r——— ‘ _ I‘— THE iNFLUENCE OF VARYiNG DEGREES OF H‘ffiRTH‘fachISM ON SEMEN PR3 UCTION N THE DOMESTEC FDWL Thesis 4501‘ the Degree of M. S. MICHEQZAN SUSIE COLLEGE Mbarm Mex-Marita "Néiwsri‘h W43 THESlS This is to certify that the thesis entitled The Influence of Varying Degrees of Hyperthyroidism on Semen Production in the Domestic Fowl presented by Alberto Monteiro Wilwerth has been accepted towards fulfillment of the requirements for Magma in Physiology _ Z . d QM --_.__ Major professor Date__M_s1_2L_lege_ _- M-796 THE INFLUENCE OF VARYING DEGREES OF HYPEHTBYROIDISM ON SEMEN PRODUCTION IN THE DOMESTIC FOWL By ALBERTO MONTEBIRO gngssTH A THESIS Submitted to the School of Graduate Studies of Michigan State College of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Physiology and Pharmacology 1948 THES‘S ACKNOWLEDGMENT The author wishes to express his sincere appreciation to Professor 5. P. Reineke, Department of Physiology and Pharmacology, Michigan State College for his wise orientation in the course of this work and for his critical readings of the manuscript; to Professor B. B. Roseboom, Head of the Department of Physiology and Pharmacology of Michigan State College for the facilities given to us in conducting this experiment; to L. F. Wolterink, Associate Professor of the Department of Physiology and Pharmacology, Michigan State College, for his valuable suggestions; and to Professor J. A. Davidson of the Poultry Department for making the animals available for this work. Be especially wishes to express his appreciation to Odette F. Wilwerth, his wife, both for technical assistance and for her continued encouragement during the course of this work. 1303185 TABLE OF CONTENTS lI.J'l.-IRODUCTIONOO000......O...0..OOOOOOOOOOOOOOOOOOO... 1 RELATIONSHIP OF THE THXHULD TU FESTILITY IN DOMESTiC ANIMALS............................................. 2 REVIEW OF LITSHATUHE Relationship of the Thyroid to Reproduction: Cattle.......................................... 3 Sheep and Goats................................. 4 FOWISOOOOO0....OOOOOOOOOOOOODOOOOOOOO00.0.0.0... 5 Effect of Thyroid Therapy on Testes and Sperm PrOductiOnOOOOOOO0.0000000000000.00.000.000.000. 7 Effect of Hypothyroidism on Testes and Sperm Production...................................... 8 EXPESIMENTAL PROCEDURE Birds and Material.............................. 10 General Plan.................................... 12 Semen Volume................................ 12 Sperm Concentration......................... 15 Total Number of Sperm....................... 13 EXPEEIMSNTAL RESULTS............................... 14 Semen Volume.................................... 14 Sperm Uoncentration............................. 26 Total Number of Sperm........................... 36 DISCUSSION AND SUMMAhY............................. 45 CONCLUSIONS........................................ 47 REFEREIJUES.000......IO...0.0...OOOOOIOOOOOOOOOOOOOOO 48 INTRODUCTION It is well-established that the thyroid gland has an important function in life processes. Thyroid removal will prevent maturation changes such as the metamorphosis of tadpoles as well as the growth and sexual development of higher animals. It will also depress many productive processes such as egg production in chickens and milk pro— duction in cows. In adult animals thyroid deficiency often results in decreased fertility. In recent years a great deal of work has been done in animals that were rendered hypothyroid either by means of thyroidectomy or administration of the goitrogenic drugs. Most authors are in agreement that the decrease in metabolism observed in the hypothyroid state is accompanied by a depression of growth, milk production, sexual activity and many of the general body processes. These functions can be improved, on the other hand, by the administration of thyroid substance. Modern investigations of thyroid function had their inception with the observations of Gull (1874) and Reverdin (188z) in women with spontaneous atrOphy of this gland. The discovery of iodine in the thyroid by Bauman (1895) lead to the use of this element in the correction of cer— tain types of thyroid abnormalities. Since that time a very extensive literature has been developed on both the clinical aspects and the fundamental nature of the physiology of the thyroid gland. A new impetus to thyroid investigations was produced by the discovery of the goitrogenic effects of sulfaguana- dine (MacKenzie, MacKenzie and McCollum, 1941), and the ser- ies including thiourea and thiouracil (MacKenzie and MacKenzie, 1943; Aswood et a1, 1945). With the develOp- ment of thyroidally active iodinated casein by Reineke and Turner (1943), it became possible to vary the level of thyroid function of domestic animals at will, simply by the suitable administration of a goitrogen to produce the‘ hypothyroid state or therprotein to create the desired level of hyperthyroidism. So far as the domestic animals are concerned the principal emphasis thus far has been placed on the study of such functions as milk production, egg production and growth. Little attention has been given to the possible relationships of the thyroid hor- mone and the reproductive functions until quite recently. The investigations to be reported represent a study of the effect of.different degrees of hyperthyroidism on semen production in the domestic fowl. This work is a Continuation of the investigations started by Martinez (1947). Relationship of the Thyroid to Fertility in Domestic Animals Many clinical reports are available that indicate that hypothyroidism in man is accompanied by reproductive disorders. In the male, deficient Spermatogenesis and a decline in libido are often observed. In the female, irregular menstrual cycles and lowered fertility accom- pany this condition. Many cases have been reported in which such conditions are corrected by thyroid therapy. It is only during the last few years that any attention has been given to the panning role of the thyroid in the reproductive processes of domestic animals. This work has been confined principally to cattle, goats and birds. Relationship of theAThyroid to Reproduction in Cattle: Petersen, Spielman, Pomeroy and Boyd reported that when a male Jersey was thyroidectomized at four months of age a complete suppression of libido occurred. However, semen that was obtained by rectal manipulation of the ampulla was used successfully for the insemination of cows. Oral administration of desiccated thyroid restored normal activity and sexual behavior. With the development of therprotein, Reineke and Turner (1945) and Turner (1945) suggested the possible use of this product to correct breeding deficiencies in bulls that become deficient in breeding performance due to subnormal thyroid activ;ty. Heineke (1946) reported the effects of feeding thyro- protein to 14 bulls which had unsatisfactory breeding records. Definite improvement in vigor and libido was observed in 10, the time required for an Observable effect to occur ranging from 7 to 40 days. Definite evidence of improvement in the conception records was observed in four cases. Results of thyroidectomy on reproduction in the fe- male bovine were reported by Spielman et a1 (1945). Four thyroidectomized cows failed to manifest the normal physical signs of estrous. By rectal palpation it was established, however, that normal ovulatory cycles were occurring. Artificial insemination of the cows with semen from a thyroidectomized bull resulted in conceptions in all of the animals. It would appear therefore that the most significant effect of hypothyroidism on the repro- ductive functions is a pronounced decrease in sex interest in both the male and female. However, even though the thyroidectomized bull employed in these cases produced viable semen the data presented were too limited to judge the intensity of spermatogenesis. Relationship of the Thyroid to Reproduction in Sheep and Qpappz It is well known that sheep and goats are seasonal breeders, exhibiting little or no sexual activity during the summertime and resuming their breeding cycles in the Fall. Berliner and Warburton (1957) reported that Shrop- shire rams produced semen of poor quality during the hot months of summer but only the poorer Hampshires declined noticeably during this period. Hampshire rams that were lthyroidectomized in the Fall showed a deficiency of sperm production similar to that observed in intact Shropshires during the Summer. Sperm production was improved in both conditions by the injection of thyroxine. Turner, Mixner and Beineke (1943) reported definite improvement in the semen production of a ram which had good sex drive but was deficient in spermatogenesis. A male Toggenburg goat that showed a definite lack of sex interest was fed thyroprotein at the rate of one gram daily. This animal showed a considerable increase in sexual vigor and settled five out of six females that were bred to him. A quite extensive study on the relationship of seasonal temperature changes and the thyroid to sperma- togenesis in rams was reported by Bogart and Mayer (1946 a, b). The administration of thyroxine or thyroprotein largely prevented the usual Summer decline in spermato- genesis of rams. Changes in semen quality similar to those resulting from high environmental temperatures were induced during the breeding season.by the administration of thiouracil. These changes were also counteracted by the administration of thyroprotein. Belationship of the Thyppid to Reproduction in Fgwlgp In recent years a considerable amount of work has been done on the possible relationship of the thyroid to re- production in both hens and cocks. However, there have been large differences in the conditions of management and the dosage of thyroidal substance used in the various .1aboratories. Inasmuch as the level of dosage of this type of substance is quite critical, the variance in re- sults from several of the laboratories may be explained upon this basis. Hens: In a very famous investigation Crew (1925) reported the rejuvenation of seven hens, 5 to 8 years of age when they were fed desiccated thyroid daily for six months. The principal changes were the development of new plumage characteristic of younger fowls, improve— ment of the head furnishings and an increase in egg pro- duction. Some improvement in the head furnishings and the egg production of hens was reported by Greenwood and Blyth (1942) following treatment with thyroid substance. With the development of synthetic thyroprotein, Turner, Irwin and Reineke (1945), and Turner, Kempster, Hall and Reineke (1945) reported an increase in the egg pro- duction of White Leghorn hens when thyroprotein was fed continuously at the level of 10 gm. per 100 lbs of feed. The greatest effect observed was a tendency toward main- tenance of the intensity of egg production during the hot months of summer. A similar maintenance of egg production in these same birds when continued on therprotein feeding in their fourth and fifth laying years has been reported by Turner, Kempster and Hall (1946) and Kempster and Turner (1947). Cole and Butt (1927) reported that desiccated thy- roid when fed at the rate of 59 mgs. per pound of live weight for a period of six weeks had no effect on egg production. The difference between this and results cited earlier can probably be eXplained by differences in dosage. That the thyroid secretion is involved in egg pro— duction in the hen is also indicated by the reports of Winchester (1940) and Blivaiss and Domm (1942). In the first report egg production was reduced markedly by thyroidectomy and in the second the thyroidectomized hens were not known to lay any eggs up to 8 months of age. Egg production was restored (Winchester, 1940) following the injection of thyroxine. Effect ofwThyroid Thprapy on Tespgs and Sperm Production: Crew (1925) reported the rejuvenation of five cocks, 5 to 8 years of age that were fed desiccated thyroid. This report is based principally on the physi- cal appearance of the birds, however. Jeep (1955) observed that when .25 to 1.0 gram of desiccated thyroid was fed daily to Mallard drakes during late Winter and early Spring both the size of the testes and spermatogenesis increased much more than in normal drakes subjected to increased light exposure alone. Aron and Benoit (1954) also observed sexual stimulation of immature male ducks when they were fed thyroid tissue or were injected with thyroxine. When fed a large dose of desiccated thyroid (100 mg. daily) white Leghorn cockerels showed an increase in the rate of seasonal decline in amount of semen produced (Titus and Burrows, 1940). On the other hand, Hayes (1948) observed that during the period from January 12 to March 21 the seasonal decline in fertility of cocks could be prevented either by the administration of small doses of thyroxine or the use of supplementary lighting. Martinez (1948) observed a definite increase in semen volume, Sperm concentration and total number of Sperm produced by male fowls that were fed thyroprotein at the rate of 18 gms. per 100 pounds of feed. Lower dosage levels were ineffective. Effect of Hypothyroidism on Testgs and Spgrm Production: Following thyroidectomy of male chickens and ducks, Benoit and Aron (1954) reported delayed testicular growth in young birds of both Species. In older birds thyroidectomy resulted in a pronounced decrease in size of the testes. Thyroidectomy also reduced considerably the testicular growth in immature ducks exposed to the stimulating action of supplemental lighting (Benoit, 1956). Spermatogenesis was affected similarly (Benoit, 1957a), and development of the penis of thyroidectomized ducks was inferior (Benoit, 1957b, 1957c). Greenwood and Chu (1959) and Blivaiss and Domm (1942) reported that when Brown Leghorn cockerels were thyroidectomized there was a pronounced reduction in the weight and histological development of the testes as well as de- vficient Sperm production. Shaffner and Andrews (1948) fed rations contain- ing 2/10 and 5/10 per cent thiouracil in order to sup- press thyroid function. Neither sperm concentration, total number of spermatozoa or methylene blue reduction time was affected by the feeding of thiouracil. Initial motility was lowered considerably at the 5/10 per cent level. Sperm survival time at 4° C. was decreased by both dosages. Actual tests of fertility were made by inseminating hens with semen from the control and exper- imental birds. Both levels of thiouracil caused a signi- ficant reduction in fertility of the males. With the foregoing work as a background it seemed of considerable interest to carry on further investiga- tions on the possibility of influencing the intensity of spermatogenesis in adult male chickens by properly regulated dosages of therprotein. With this in mind, an attempt has been made to establish the effects of a relatively low dosage level that would be eXpected to stimulate spermatogenesis to some extent, and also to determine whether excessive levels given under the same conditions would cause inhibitory effects. EXPERIMENTAL PROCEDURE Birds and Material Fifteen 8-month-old hhode Island Red roosters obtained from the Poultry Department of Michigan State College, were selected for the experiment from a group of about 28, after a preliminary training period of 5 months. They were placed in individual cages on Octo— ber 18, 1947 and fed a basal diet which was compounded as shown in 690 400 300 200 so so 40 50 50 100 so 12 ___8 2000 During the accompanying table: lbs. lbs. lbs. lbs. lbs. lbs. lbs. lbs. lbs. lbs. lbs. lbs. lbs. lbs. Corn Meal Ground Oats Bran Middlings 17% dehydrated alfalfa meal Meat scraps Dry milk Fish meal Soybean meal Oyster shell flour Steamed bone meal Salt Fish liver 011 (400 A., 2000 D.) the preliminary period semen was collected from the roosters three times a week, by the method of Burrows and Quinn (1957). After this preliminary period that extended for three months, in order to allow the roosters to mature and give uniform samples, the roosters were distributed into three groups of 5 birds each on the basis of the average semen production. One other category of birds used consisted of 8 old cocks, about 21 months old, selected from a group -10- of 12, on the same basis as for the young birds; these old roosters formed two groups of four birds each, and they received the ration described above. The room temperature was maintained at about 65°F. during the period of the experiment, that is, from October till March, and the birds received artificial light 24 hours a day throughout the experimental period. Our experiment started on.January 9 and lasted until March 18 with the young birds; with the older ones, we finished on February 27. From January 28 each group received a different treatment. The drug we used, Thyroprotein, a highly active iodinated protein (Protamone), was supplied by Cerophyl Laboratories, Inc., Kansas City, Missouri. This drug was given to the birds together with the feed, being carefully mixed in it, having in account the small amounts used and required to stimulate feather growth and egg production as reported by Turner, Irwin and Reineke (1945) and to improve semen production as re- ported by Martinez (1947). Thyroprotein was added to the ration of the experi- mental birds as indicated below: Young roosters: Group A - Normal control: this group received only the basal mash, throughout the period of experi- ment; Group B - Received thyroprotein at the level of 18 grams -11... per 100 pounds of feed, that is, 0.04% through- out the period; Group C - Received thyroprotein at the rate of 56 grams per 100 pounds of mash, or 0.08%. This group in the two last weeks of the experiment re- ceived four times the level of group B, or 72 grams per 100 pounds of feed. Old roostepp Group D - Normal control: treated in the same way as the normal young roosters; Group E - Received thyroprotein at the level of 0.04%, throughout the period of experiment. All birds received the mash "ad libitum"; the food was supplied twice a day and they had running tap water at all times. General plan Semen was collected from the birds three times a week, with the purpose of determining the possible variations in semen volume and sperm concentration under the influence of thyroprotein. To collect the samples we used the method of Burrows and Quinn (1957). After the birds were well trained they produced quite constant amounts of semen. The samples were collected at intervals of 48 hours as recommended by Parker, McKenzie and Kempster (1942). a. Semen volume The volume was determined in a 1 ml. tuberculin -12- syringe and the measurements were made as accurately as possible, giving us readings in hundredths ml. We collected samples every week, throughout the period of experiment, two or three times a week. An interval of forty-eight hours was observed between the collections. b. Sperm concentratigp — To determine the Sperm concentration we employed the hematocytometer used in counting red blood cells. To kill the sperm in order to avoid troubles in count- ing, we added a few drops of alcohol to the Tyrode's solution used to dilute the semen; we have used too, with advantage, the following solution for dilution: Methylene blue - 50 ml. KOH 2% - 60 ml. Acetic acid - .5 ml. The dilution of the semen was 1:200, that is the undiluted semen was taken up to the 0.5 mark in the mix- ing pipette for red blood cell counts and the dilute solution was taken up to the 101 mark. From each sample we did two and sometimes three counts and the averages obtained were taken as our values for sperm concentration per cubic millimeter. c. Total number of sperp The total number of sperm in the ejaculate is based on the concentration per cubic millimeter and the volume; knowing these data we just multiply concentration by volume in order to get the total number of Sperm. -15- EXPERIMENTAL RESULTS We collected control semen samples for this experi- ment from January 9 to January 28, 1948, from all groups of birds. On January 28 we started giving to the young roosters thyroprotein as follows: to group B we gave 0.04% in the feed, throughout the period of experiment, that is, until March 18: to group C we gave 0.08% in the feed until March lst, when we started giving twice this dose (0.16% in the feed), for the rest of the period. In the old roosters we had just two groups: group D that was the normal control and group m, receiving thyro- protein at the 0.04% level. The period of experiment for these two groups (D and E) started on January 28 and lasted until February 27. All the birds were in good health throughout the period of eXperiment and no variation in semen production due to sickness or any other factor was observed. Our results will refer to the groups of young and old roosters separately. Semen Volume Xpunggroospeps - In this category we had three different groups, each receiving a different treatment during the period of experiment. Group A.« This group was the normal control in this cate- gory and did not receive any therprotein throughout the -14- period of experiment. The averages obtained during this period were be- tween 0.40 and 0.68 ml. This group was quite constant in giving samples. The highest volume observed was 0.85 and the lowest was 0.22 ml. The greater part of the data fell between 0.50 and 0.70 ml. The results obtained from this group are shown in table I, and a comparison with the other groups will be given in table IV. TABLE I - The Semen Volume of Control Roosters. ._f SEMEN VOLUME in ml. Date Birds 1 2 5 4 5 Average__ Jan. 9 0.50 0.80 0.75 0.50 0.85 0.68 14 0.45 0.64 0.58 0.20 0.61 0.50 16 0.57 0.59 0.80 0.55 0.44 0.51 19 0.57 0.56 0.70 0.71 0.85 0.68 25 0.52 0. 66 0.46 0.58 0.60 0.52 26 0.41 0. 66 0.70 0.65 0.45 0.57 28 0.60 0. 67 0.65 0.74 0.84 0.69 50 0.42 0.67 0.65 0.66 0.48 0.57 Feb. 2 0.57 0.49 0.71 0.55 0. 58 0. 58 4 0.42 0.77 0.66 0.67 0. 65 0. 65 9 0.50 0. 70 0.65 0.59 0. 45 0.54 11 0.25 0. 61 0.22 0.52 0. 58 O. 59 15 0.59 0.55 0.41 0.57 0.66 0.52 16 0.58 0.60 0.45 0.55 0.60 0.52 18 0.40 0.42 0.50 0.55 0.66 0.45 20 0.26 0.60 0.55 0.58 0.59 0.40 25 0.27 0.60 0.40 0.50 0.40 0.45 27 0.50 0.50 0.58 0.51 0.45 0.47 Mar. 1 0.40 0.60 0.40 0.50 0.50 0.48 5 0.58 0.50 0.45 0.50 0.60 0.48 5 0.49 0.55 0.47 0.51 0.49 0.46 8 0. 55 0.55 0.55 0.45 0.55 0. 55 10 0. 25 0.75 0.45 0.50 0.50 0. 49 18 0.46 0.74 0.45 0.45 0.47 0. 51 Group B - This group received the lowest dose of thyro- protein we have used in this eXperiment - 0.04% of therprotein in the feed. Feeding was begun on Jan- uary 28 and continued for 50 days until March 18. The volume from this group was not increased when compared with the preliminary period (between January 9 to Jan- uary 27), but the averages during the period of experi- ment were more constant and the semen voluem level was maintained just about the same throughout the period and at a more constant and higher level on the average than in Group A. The highest value was Olggyml. and the low— est was OL§§Hml.; mostcfi'the samples were between 0.50 and 0.70 ml. The data obtained from this group are shown in table II and a comparison will be made with the other two groups in table IV. -16- TABLE II - The effect of thyroprotein on semen production of young roosters when given as 0.04% of the ration SEMEN VOLUME in m1. Birds Date *= 9 10 11 12 13 Average Jan. 9 0.95 0.67 0.43 0.45 0.65 0.63 14 0.76 0.70 0.62 0.55 0.67 0.66 16 0.57 0.48 0.48 0.45 0.40 0.48 19 0.75 0.60 0.70 0.67 0.46 0.64 23 0.65 0.50 0.45 0.61 9.57 0.55 26 0.38 0.61 0.40 0.95 0.47 0.56 28* 0.65 0.42 0.46 0.61 0.65 0.56 30 0.56 0.57 0.62 0.69 0.75 0.64 Feb. 2 0.55 0.57 0.50 0.60 0.85 0.61 4 0.67 0.66 0.50 0.77 0.58 0.64 9 0.59 0.75 0.40 0.64 0.38 0.55 11 0.65 0.60 0.41 0.46 0.52 0.53 13 0.46 0.44 0.40 0.51 0.44 0.45 16 0.70 0.70 0.47 0.60 0.60 0.61 18 0.68 0.70 0.48 0.65~ 0.70 0.64 20 0.65 0.68 0.55 0.60 0.65 0.62 25 0.70 0.65 0.50 0.65 0.70 0.64 27 0.72 0.56 0.60 0.45 0.78 0.62 Mar. 1 0.60 0.55 0.61 0.50 0.75 0.60 3 0.70 0.67 0.45 0.57 0.60 0.60 5 0.41 0.59 0.38 0.30 0.75 0.49 8- 0.60 0.85 0.53 0.40 0.65 0.61 10 0.60 0.70 0.58 0.30 0.65 0.56 18 0.40 0.85 0.45 0.25 0.80 0.55 *Started receiving 0.04% of thyroprotein. Group C - This group received two different doses of therprotein throughout the period of experiment; from January 28 until February 29 it received 0.08% of thyro- protein in the feed; from March 1 until the end of the experimental perion (March 18) it received twice this dose, that is, 0.16% of therprotein. The semen volume in this group was not so constant 1n as in‘the two others. By the end of the experiment the average volume had gone down about 50%. The highest volume checked was 1.37 ml. and the low- est volume was 0.18 ml. The data in the first half of the 'experiment were about the same as the other two groups; between 0.40 and 0.60 ml. Later the volumes declined rather markedly. In table III are shown the volumes ob- tained from group C and a comparison with the two other groups will be given in table IV. -15- TAELE III - The effect of thyroprotein on semen production of young roosters when given as 0.08 and 0.16 per cent of the ration. SEMEN VOLUME in ml. “r-— BiTQ§ _ Date 20 21 22 23 24 Average Jan. 9 0.30 0.35 0.95 0.35 0.33 0.34 14 0.55 0.30 0.57 0.40 0.95 0.31 13 0.30 0.45 0.53 0.39 0.77 0.59 19 0.41 0.4 0.79 0.49 0.91 0.31 23 0.40 0.2 0.43 0.39 0.30 0.42 23 0.42 0.40 0.44 0.70 1.37 0.37 23* 0.43 0.2 0.42 0.30 1.07 0.52 30 0.32 0.31 0.32 0.35 0.94 0.37 Feb. 2 0.57 0.39 0.39 0.33 0.55 0.54 4 0.57 0.45 0.43 0.47 0.35 0.51 9 0.30 0.35 0.43 0.47 0.35 0.51 11 0.32 0.40 0.32 0.44 0.70 0.50 13 0.33 0.31 0.30 .29 0.45 0.3 13 0.33 0.25 ,0.45 0.40 0.35 0.49 19 0.45 0.50 0.24 0.45 0.30 0.45 20 0.52 0.32 0.22 0.20 0.34 0.39 25 0.30 0.40 0.20 0.25 0.50 0.37 2 0.53 0.40 0.33 .0.39 0.30 0.39 Mar. 1** 0.52 0.35 0.30 0.45 0.30 0.39 3 0.50 0.40 0.40 0.39 0.33 0.41 5 0.45 0.33 0.45 0.39 0.32 0.5. 8 0.50 0.40 0.4 0.30 0.20 0.33 10 0.59 0.47 0.2. 0.33 0.13 0.37 18 0.35 0.40 0.40 0.13 0.33 0.45 *Started receiving 0.08% of thyroprotein. ** Started receiving 0.16% of thyroprotein. Table IV - Summarizes the effect of different levels of thyroprotein on semen volume of the three groups of young birds. These data are the average obtained from those groups shown in the three first tables. The semen volume of the first three groups is com— pared in figure I. Group B, receiving thyroprotein as 0.04 per cent of the feed, showed more constant and high- er average values than the control group. The semen volume of group C was less constant and showed a marked decline when the dose of thyroprotein was increased to 0.16 per cent of the feed. TABLE IV - Summary of the effect of different levels of thyroprotein on semen production of young roosters. SEMEN VOLUME in ml. ,1 _EQBP§_ ___. Date A B C Jan. 9 0.68 0.63 0.64 14 0.50 0.66 0.61 16 0.51 0.48 0.59 19 0.68 0.64 0.61 23 0.52 0.55 0.42 2 0.57 0.56 0.67 28 0.69 0.56* 0.52** 30 0.57 0.64 0.67 Feb. 2 0.58 0.61 0.54 4 0.63 0.64 0.51 9 0.54 0.55 0.56 11 0.39 0.53 0.50 13 0.52 0.45 0.34 16 0.52 0.61 0.48 20 0.40 0.62 0.38 25 0.43 0.64 0.37 27 0.47 0.62 0.39 Mar. 1 0.48 0.60 0.39*** 3 0.48 0.60 0.41 5 0.46 0.49 0.39 8 0.53 0.61 0.36 10 0.49 0.56 0.37 18 0.51 0.55 0.36 g§tarted receiving thyroprotein 0.04 per cent in the ration **Rece1ved 0.08 per cent of therprotein until February 29 ***The dose of thyroprotein was increased to 0.16 per cent. 0.7 ‘1 A ‘ . , .\ 0.6 ‘ . ‘ ‘V\ ~ 1. 0 V V - ‘ 0.5. , _ lu ’ ’4 3 ’ \V . 5 3414. ~ X E l“ c z 0.3. '44 v) 0.2. L- T”. 0.04): , B __ Ira 0.007.,c ITP.0.IO‘Z,C 0 10 20 :0 40 50 60 70 DAYS 0N EXPERIMENT Figure I - The effect of therprotein on semen volume of young roosters when given as 0.04, 0.08 and 0.16 per cent of the feed. -21- 01d roosters In this category of birds we worked with two groups: group D, the normal control, did not receive any treat- ment, and group B received thyroprotein at the level of 0.04% in the feed. Group D - This group did not receive any treatment throughout the period of experiment which started on January 9 and went on until February 27. The highest semen volume checked was 0.80 ml. and the lowest was 0.20 ml. Most of the values were between 0.30 ml. and 0.55 ml. The results obtained from this group are shown in table V and a comparison with group E will be shown in table VII. Table V — Semen volume of control roosters. SEMEN VOLUME in ml. Birds Date 41 42 43 45 Average Jan. 9 0.40 0.50 0.75 0.40 0.51 14 0.27 0.19 0.37 0.43 0.31 13 0.25 0.34 0.35 0.43 0.42 19 0.22 0.23 0.53 0.49 0.38 2 0.24 0.37 0.48 0.45 0.39 23 0.55 0.49 0.27 0.90 0.53 28 0.39 0.50 0.45 0.57 0.49 30 0.49 0.55 0.44 0.33 0.53 Feb. 2 0.29 0.39 0.32 0.33 0.40 4 0.31 0.39 0.24 0.40 0.41 9 0.29 0.45 0.41 0.30 0.44 11 0.23 0.40 0.45 0.51 0.40 13 0.32 0.20 0.13 0.55 0.31 13 0.50 0.32 0.30 0.35 0.44 19 0.55 0.25 0.20 0.5 0.39 20 0.30 0.30 0.20 0.5 0.39 25 0.39 0.30 0.20 0.55 0.33 27 0.32 0.40 0.22 0.50 0.33 Group E - This group received therprotein at the dose of 0.04”. Feeding was begun on January 28 and lasted for 30 days until February 27. The highest volume check- ed was 0.85 ml. and the lowest was 0.18 ml. The averages were between 0.30 and 0.62 ml. and most of the volumes checked were between 0.30 and 0.55 ml. In table VI are shown the individual volumes ob- tained in this group. A comparison with group D will be made in table VII. 'TABLE VI - The effect of thyroprotein on semen produc- tion of old roosters when given as 0.04 per cent of the ration. SEMEN VOLUME in m1. D t __ _____ Birds __ a e - 56 _ 57 38 g__391_Axerage Jan. 9 0.46 . 0.48 0.75 0.23 0.48 14 0.25 --- 0.58 --- -~- 16 0.28 0.33 0.40 0.18 0.30 19 0.48 0.37 0.46 90.25 0.39 23 0.34 0.39 0.56 0.21 0.37 26 0.37 0.22 0.78 0.42 0.45 28* 0.40 0.57 0.70 0.31 0.50 30 0.47 0.57 0.75 0.32 0.53 Feb. 2 0.27 0.40 0.58 0.41 0.41 4 0.31 0.64 0.84 0.47 0.56 9 0.30 0.30 0.55 0.27 0.35 11 0.35 0.35 0.79 0.68 0.62 13 0.25 0.57 0.33 0.23 0.34 16 0.38 0.27 0.70 0.30 0.41 18 0.30 0.40 0.71 0.40 0.45 20 0.22 0.55 0.56 0.50 0.46 25 0.25 0.45 0.61 0.55 0.46 27 0.29 0.57 0.81 0.65 0.58 ;§tarted receiving 0.04% of thyroprotein in the feed. -25- Table VII - In this table are summarized the averages of semen volumes obtained from groups D and E. Group D did not receive any treatment and group E received 0.04% of thyroprotein in the feed, during 30 days. A graph will show more clearly the variations in the semen volume in these two groups. (Figure II). The average values of group B, that received thyroprotein as 0.04% of the feed were increased near the end of the experiment, when compared with the control group D. TABLE VII - Summary of semen volume of old roosters. ___A__A A SEMEN VOLUME in cc. Groups Date D - E Fw Jan. 9 0.51 0.48 14 0031 ""'- 16 0.42 0.30 19 0.58 0.39 23 0.38 0.37 26 0.53 0.45 28* 0.48 0.50 30 0.53 0.53 Feb. 2 0.40 0.41 4 0.41 0.56 9 0.44 0.35 11 0.40 0.62 13 0.31 0.34 16 0.44 0.41 18 0.39 0.45 20 0.38 0.46 25 0.36 0.58 27 0.36 0.58 *Group E started receiving 0.04% of thyroprotein in the feed. .. 24- 6L6. E' 045 k: k 13 -4 (3(14. 3 E: .D 3 043 ‘u U) .9 I“ L_ {TR 0042, E 0 1'0 20 :0 , 40 50 0,4125 0N EXPERIMENT Figure 11 - The effect of thyroprotein when given as 0.04% of the feed, on semen volume of old roosters. -25- §perm Concentration_ Young r0049; Group A - This group was our normal control and did not receive any treatment during the period of experiment. Sperm concentration was checked every week from January 23 until March 18. The data obtained in this group were quite constant, the highest average value being 2,200,000 sperm per cubic millimeter and the lowest average value being 1,854,000 sperm per cubic millimeter. The highest individual value was 2,390,000 and the low- est was 1,670,000 sperm per cubic millimeter. The results obtained from this group are seen in table VIII. In table XI a comparison will be made with the other two groups - B and C. TABLE VIII - The sperm concentration of the control group of young roosters. 'V—w NUMBER OF SPERM IN MILLIONS PER cu. mm. Birds Date 1 2 5 4 5 Average Jan. 25 1.820 1.780 1.760 1.980 1.950 1.854 26 2.050 2.070 2.150 2.110 2.200 2.112 50 1.670 2.180 2.000 1.980 1.820 1.950 Feb. 4 2.040 2.250 2.180 2.220 2.170 2.172 15 2.160 2.020 2.060 2.590 1.920 2.110 20 2.050 2.010 1.890 2.210 1.980 2.024 27 1.950 2.020 2.190 2.520 1.870 2.066 Mar. 8 2.090 2.290 2.040 2.280 2.250 2.190 10 2.050 2.500 2.060 2.550 2.280 2.200 18 2.010 2.260 2.180 2.210 2.090 2.150 Group B - This group received 0.04% of thyroprotein in the feed throughout the period of experiment, from January 28 to March 18. Two weeks after receiving thyro- protein the birds showed a little improvement in sperm concentration as can.be seen in table IX. The highest value for this group was 2,590,000 Sperm per cubic millimeter and the lowest was 1,790,000 Sperm per cubic millimeter. The highest average value was 2,390,000 and the lowest was 1,888,000 Sperm per cubic millimeter. The data concerning this group are given in table IX and the averages are put together in table XI, for comparison. TABLE IX - The effect of 0.04 per cent of thyroprotein on sperm concentration in young roosters. NUMBER OF SPERM IN MILLIONS PER cu. mm. D t _, Birds a e 9 10 11 12 13 Average Jan. 23 1.860 1.900 1.790 1.940 1.950 1.888 26 2.020 2.060 2.110 2.180 2.130 2.100 30 1.890 2.220 2.410 2.320 2.040 2.176 Feb. 4 2.160 2.670 2.180 2.310 1.860 2.236 13 1.970 2.140 2.440 2.550 1.910 2.202 20 2.210 2.380 2.310 2.380 2.250 2.306 27 2.110 2.280 2.570 2.110 2.050 2.224 Mar. 8 2.100 2.250 2.500 2.450 2.200 2.300 10 2.200 2.300 2.590 2.490 2.370 2.390 18 2.210 2.250 2.450 2.430 2.410 2.350 Group B received -27... 0.04% of thyr0protein in the feed; feeding was begun on January 28 and continued for 50 days. Group C - This group received two different levels of therprotein, during the period of experiment. From January 28 until February 29 they received 0.08% of thyroprotein in the feed; from March 1 until the end of the experiment they had twice this dose, that is, 0.16% of thyr0protein. The highest average value was 2,108,000 Sperm per cubic millimeter and the lowest was 1,374,000 Sperm per cubic millimeter. The highest individual value was 2,500,000 and the lowest was 1,310.000 Sperm per cubic millimeter. There was a marked decrease in sperm concentration at the end of the experiment due probably to the quite heavy dosage of therprotein. The data obtained from this group are Shown in table X and the averages given in table XI for compari son with the other groups. TABLE X - The effect of thyroprotein on sperm concen- tration of young roosters when given as 0.08 and 0.16 per cent of the ration. NUMBER OF SPERM IN MILLIONS PER cu. mm. Date Birds _ Jan. 23 1.680 1.970 1.750 1.730 2.040 1.834 26 1.990 2.030 1.960 2.170 2.210 2.072 30 1.890 1.850 2.500 2.330 1.950 2.104 Feb. 4 1.180 2.170 2.290 2.450 2.310 2.080 13 1.820 1.930 1.360 1.770 1.890 1.754 20 1.830 2.050 1.800 1.960 1.980 1.880 27 1.990 1.750 1.570 1.820 1.880 1.802 Mar. 8 1.600 1.560 1.530 1.690 1.770 1.630 10 1.700 1.650 1.570 1.690 1.590 1.640 18 1.390 1.360 1.440 1.370 1.310 1.374 Received therprotein as 0.08% of the feed from Jan- uary 28 until February 29; and 0.16% from March 1 to marCh 18 0 Table XI will Show a summary of the average values obtained from the birds in the three groups and it shows the effect of different dosages of thyroprotein on Sperm concentration. A graph of this data will show these variations more clearly (Figure III). Spermatogenesis was influ- enced by the administration of thyroprotein at differ— ent levels. When given as 0.04% of the feed (group B) sperm concentration was increased markedly. When given at higher levels, as 0.08% and 0.16% of the feed (group C), a marked decline in sperm concentration was observed, due probably to the depressing effect of ex- cessive thyroprotein on the gonads. Sperm concentration of the control group (group A), was quite constant throughout the experimental period. TABLE XI - Summary showing the effect of different levels of thyroprotein on sperm concentration of young roosters. NUMBER OF SPEKM 1N MILLIONS PEfi cu. mm. _. Groups __ Date A B 0 Jan. 23 1.854 1.888 1.834 26 2.112 2.100 2.072 30 1.930 2.176 2.104 Feb. 4 2.172 2.236 2.080 13 2.110 2.202 1.754 20 2.024 2.306 1.880 27 2.066 2.224 1.802 Mar. 8 2.190 2.300 1.630 10 2.200 2.390 1.640 18 2.150 2.350 1.374 Group B received 0.04% of thyroprotein in the feed from January 28 to March 18. Group C received 0.08% of thyroprotein from January 28 until February 29; and 0.16% from March 1 to March 18. -50.. 2‘ Q l / 2.4. B .32- 2 ,f‘\ 'A 0 r R \ ‘ 2.0.] \ Q I 2 , \ \' A / \ {4,1,0 \ , ‘x 2 V \. Q \ U \ k Q“ Kl Q. v; ~. «L 0’ '3 1.2- . 7/2 0.042 , a KIT/10082.6 TR 0.167.,6 0 f 10 20 30 40 50 60 DA VS 0N EXPERIMENT Figure III - The effect of thyroprotein on sperm con- centration of young roosters when given as 0.04, 0.08 and 0.16 per cent of the feed. -51- 01d Roosters Group D - This was the control group and did not receive any treatment during the period of experiment. The averages The for this sperm per cubic millimeter. was 2,057,000 and the lowest was 1,547,000 Sperm per cubic millimeter. are given in table XIV with group 5. data obtained are shown in table XII and the highest individual sperm concentration value group was 2,280,000 and the lowest was 800,000 The highest average value TABLE XII - The sperm concentration of the control group of Old roosters. SPEhM CONCENTRATION IN MILLIONS ———r Birds Date 41 42 '43 45 Average Jan. 19 2.150 1.950 1.810 1.930 1.960 23 2.020 2.000 1.810 2.060 1.972 26 1.970 2.070 1.910 2.120 2.017 28 0.800 2.280 1.940 1.430 1.612 Feb. 2 0.910 1.810 1.900 1.570 1.547 9 2.030 2.050 1.950 2.100 2.033 13 1.800 2.120 2.050 2.260 2.057 18 1.630 2.100 1.970 2.120 1.955 27 2.030 2.050 1.740 1.810 1.907 Group E - Feeding this group with therprotein began on Ja nuary 28 and lasted until February 27, at the level of 0.04% in the feed. The highest individual value for this group was 2,360,000 Sperm per cubic millimeter and the lowest was _'ZOl 1,520,000 sperm per cubic millimeter. The highest aver- age value waS 2,145,000 and the lowest was 1,775,000 sperm per cubic millimeter. The data obtained from this group are shown in table XIII and the averages in table XIV with group D. TABLE XIII - The effect of thyroprotein on sperm con- centration of old roosters when given as 0.04 per cent of the feed. SPEhM CONCENTRATION IN MILLIONS Birds Date 36 37 38 39 Average Jan. 19 2.090 1.990 1.580 1.910 1.892 23 2.200 2.080 1.610 2.140 2.007 26 2.090 2.010 1.590 2.060 1.937 28* 1.690 2.050 1.650 1.710 1.775 Feb. 2 1.780 2.280 1.960 2.000 2.005 9 1.660 2.150 1.700 2.210 1.930 13 1.520 2.360 2.090 2.120 2.022 18 1.820 2.010 1.900 2.100 1.957 27 2.030 2.150 2.040 2.360 2.145 *Started receiving thyroprotein as 0.04% of the feed.— Table XIV shows the average values obtained from groups D and E, group D being the normal control and E receiving 0.04% of therprotein for about 30 days. The variations in Sperm concentration of these two groups is shown more clearly in graphic form in figure IV. Sperm concentration was not influenced in this category of birds as it was in young roosters. The average values from the control group and the group receiving thyroprotein as 0.04% of the feed, were about the same 0 TABLE XIV - Summary of the Sperm concentration of old roosters. SPEhM CONCENTRATION IN MILLIONS Groups Date D E .Jano 19 1.960 1.892 23 1.972 2.007 26 2.017 1.937 28 1.612 1.775 Feb. 2 1.547 2.005 9 2.033 1.930 13 2.057 2.022 18 1.955 1.957 27 1.907 2.145 Group E received 0.04% of thyroprotein in the feed, from January 28 to the end of the experiment. -34- 2.0. - - ' 2 ’1 A a V ' . I; o Elea- E K) 2 61.5- K.) 3 ct 81.4- V) I... [TR 0.00:,5 0 lo 20 3'0 40 ans 0N EXPERIMENT 1 { -.—_——-- ~7Y i Figure IV — The effect of thyroprotein when given as 0.04% of the feed on Sperm concentration of old roosters. Total Number of Sperm Group A - The data obtained from this group are shown in table XV and the average values will be put together with groups B and 0 in table XVIII. The highest individual value was 1,732 millions of sperm and the lowest was 501 millions of sperm per ejaculation. The highest average value was 1,368 millions of sperm and the lowest was 809 millions of sperm. TABLE XV - The total number of Sperm of the control group of young roosters. TOTAL NUMBER OF SPERM IN MILLIONS __ Birds Date 1 2 3 4 5 Jan. 25 946.43 1,174.80 809.60 752.40 1,338.00 26 852.50 1,556.20 1,505.00 1,571.50 946.00 50 701.40 1,460.60 1,260.00 1,306.80 857.60 F813. 4 856.80 1,752.50 1,438.80 1,487.40 1,267.10 15 842.40 1,111.00 844.60 1,362.30 1,267.20 20 527.80 1,260.00 661.50 859.80 772.20 27 576.00 1,010.00 1,270.20 1,182.20 841.50 Mar. 8 1,147.50 1,259.50 1,122.00 1,026.00 1,257.50 10 501.50 1,725.00 927.00 1,165.00 1,140.00 18 924.60 1,642.80 957.40 994.50 982.30 Group B - This group receiving 0.04% of thyroprotein throughout the experimental period showed a Sistent improvement in spermatogenesis. quite con- The highest individual value was 2,071 millions of sperm and the lowest was 607 millions of sperm per -36- ejaculation. The average values were: 1,431 millions of sperm for the highest and 990 millions of sperm for the lowest one. Table XVI shows the data obtained from this group and in table XVII will be given the average values with groups A and 0. TABLE 1V1 - The effect of of sperm of young roosters thyroprotein on total number when given as 0.04% of the feed. TOTAL NUIBTB 0F SPEEES IN MILLIONS Birds Date ‘ _ A 9 10 11 12 13 Jan. 23 1,209.00 950.00 805.50 1,183.40 1,111.50 26 767.60 1,256.60 844.00 2,071.00 1,001.10 30 1,058.40 1,265.40 1,494.20 1,600.80 1,530.00 Feb. 4 1,447.20 1,762.20 1,090.00 1,778.70 1,078.80 13 906.20 941.60 976.00 1,300.50 840.40 20 1,436.50 1,618.40 1,270.50 1,428.00 1,462.50 27 1,519.20 1,276.80 1,542.00 949.50 1,599.00 Mar. 8 1,260.00 1,912.50 1,325.00 980.00 1,430.00 10 1,320.00 1,610.00 1,502.2 747.00 1,540.50 18 884.00 1,912.50 1,102.50 607.50 1,928.00 Therprotein feeding was begun on January 28 and contin- ued to the end of the experimental period. Group 0 - The data obtained from this group that received higher levels in table XVII with groups A of therprotein than group B, are shown and the average values will be put together and B, in table XVIII, for comparison. The highest individual value was 3,027 millions of -57- Sperm per ejaculation and the lowest was 235 millions of sperm per ejaculation. was 1,409 millions of sperm and the lowest was 549 millions of Sperm. The highest average value TABLE XVII — The effect of thyroprotein on total num- ber of sperm of young roosters when given as 0.08 and 0.16 per cent of the feed. TOTAL NUMBER OF SPERM- IN MILLIONS Date Birds 20 21 22 23 24 Jan. 23 672.00 472.80 805.00 674.70 1,224.00 26 835.80 812.00 862.40 1,510.00 3,027.70 30 1,171.80 1,128.50 1,550.00 1,514.50 1,638.00 Feb. 4 672.60 976.50 984.70 1,151.50 1,501.50 13 637.00 598.30 408.00 513.00 850.50 20 951.60 656.00 396.00 392.00 1,267.20 27 1,054.70 700.00 565.20 691.60 564.00 Mar. 8 800.00 624.00 612.00 507.00 354.00 10 1,003.00 775.50 392.50 608.40 286.20 18 625.50 476.00 576.00 548.00 235.80 heceived thyroprotein as 0.08% of the feed from January 28 until February 29; and 0.16% from March 1 to March 18. Table XVIII Shows a summary of the average values ob- tained from the birds in these three groups and it shows the effect of different dosages of thyroprotein on the total number of Sperm. AA graph of these data will Show these variations in total number of sperm more clearly. (Figure V). The effect of varying levels of thyroprotein on semen -38— production is better seen in the total number of Sperm. It depends on volume and sperm concentration. In group B, that received 0.04% of therprotein in the feed we observed a quite consistent increase in the total number of sperm; in group C, receiving higher doses of therprotein, we observed a rather marked de- cline in the total number of sperm. In group A, the control group, we had quite constant values for the total number of sperm. TABLE XVIII - Summary Showing the effect of different levels of thyroprotein on total number of Sperm of young roosters. TOTAL NUMBER OF SPERM? IN MILLIONS Groups Date A B 0 Jan. 23 964.08 1,038.40 770.28 26 1,203.84 1,176.00 1,388.24 30 1,100.10 1,392.64 1,409.68 Feb. 4 1,368.36 1,431.04 1,060.80 13 1,097.20 990.90 596.36 20 809.60 1,429.72 731.12 27 971.02 1,378.88 702.78 Mar. 8 1,160.70 1,403.00 586.80 10 1,078.00 1,338.40 606.80 18 1,096.50 1,292.50 549.60 Group B received 0.0—% of therprotein in the feed from January 28 to March 18. Group 0 received 0.08 of thyro- protein from January 28 until February 29; and 0.16% from March 1 to March 18. -39- 1.4 5 [.2 '0 z A E \l 1.0 \l \ Q 5116 9: E /P\“‘\. / \ V) \ 06 r- V \VK\ 0.4 - TP. 0.04)! ,8 : {racoaxm [72 0.16%,6‘ 1' l l 7 1 1 0 10 20 JO 40 50 60 DAYS 0:" EXPERIMENT Figure V — The effect of thyroprotein on the total number of sperm of young roosters when given as 0.04, 0.08 and 016 per cent of the feed. -40- 01d noostggg Group D - The data from this group are given in table XIX and the average values will be given in table XXI with the averages from group B. The highest individual value was 1,690 millions of Sperm and the lowest was 254 millions of sperm per ejaculation. The highest average value was 1,069 millions of sperm and the lowest average value was 618 millions of sperm. TABLE XIX - The total number of Sperm of the control group of old roosters. u..." W. ——.. ' TOTAL NUMBER OF SPERM IN MILLIONS Birds Date " " ” 41 42 43 45 Jan. 19 473.00 507.00 1,013.60 926.40 23 484.80 740.00 868.80 927.00 26 1,083.50 1,014.30 515.70 1,696.00 28 312.00 1,140.00 873.00 686.40 Feb. 2 254.80 687.80 608.00 989.10 9 588.70 922.50 799.50 1,260.00 13 576.00 424.00 328.00 1,243.00 18 896.50 525.00 394.00 1,208.40 27 649.60 820.00 382.80 905.00 — '—— —-—— —— .— r——V v Group E - The data from this group are shown in table XX and the average values will be given in table XXI with group D. The highest individual value was 1,652 millions of sperm and the lowest was 380 millions of Sperm per ejaculation. The highest average value was 1,242 millions of sperm and the lowest was 675 millions of Sperm. TABLE XX - The effect of thyroprotein on total number of sperm when given as 0.04% of the ration in old roosters. TOTAL NUMBER OF SPERMS IN MILLIONS Date Birds 36 37 38 39 Jan. 19 1,003.20 706.60 726.80 477.50 23 748.00 811.20 901.60 449.40 26 773.30 442.20 1,240.20 865.20 28 676.00 1,168.50 1,155.00 530.00 Feb. 2 480.60 912.00 1,136.80 820.00 9 498.00 645.00 935.00 596.70 13 380.00 1,345.20 689.70 487.00 18 546.00 804.00 1,349.00 840.00 27 588.70 1,225.50 1,652.40 1,534.00 Thyroprotein feeding was begun on January 28 and continued to the end of the eXperimental period. Table XXI - In this table are summarized the average values of the total number of sperm from groups D and E. Group D did not receive any treatment and group E re— ceived 0.0 % of therprotein in the feed for 30 days. The variations in total number of sperm of these two groups are shown more clearly in figure VI. Feed- ing thyroprotein to old roosters for about 30 days, as 0.04% of the feed, increased somewhat the total number of Sperm, as can be seen in group E, when compared with the control group (group D). In the control group we had quite constant values for the total number of sperm. throughout the experimental period. TABLE XXI - Summary of the total number of sperm of old roosters TOTAL NUMBER OF SPEEMS IN MILLIONS Jan. 19 744.80 737.88 23 749.36 742.59 26 1,069.01 871.65 28 773.76 887.57 Feb. 2 618.80 822.05 9 894.52 675.50 13 637.67 687.48 18 762.45 880.65 27 686.52 1,242.94 Group E received thyroprotein as 0.04% of the feed Since January 28. -43- cg 4L4? 2 (3 ‘~ \1 1‘ 1.0 In S E Q 036 U3 0.5 I ‘ W. 0.04): Ill—- GROUP 6' b m 20 .30 40 DAYS 0N EXPERIMENT Figure VI - The effect of thyroprotein when given as 0.04% of the feed on the total number of sperm of old roosters. -44- DISCUSSION AND SUMMARY In this experiment we used two categories of birds; fifteen 8-month-old Rhode Island Red roosters were divided into three groups of five each, and eight old cocks about 21—month-old were divided into two groups of four each. All birds were placed in individual cages, receiv- ing the same basal diet "ad libitum", supplied twice a day, had running tap water and received artificial light twenty-four hours a day. The room temperature was main- tained at about 65°F. during the experimental period. Each group received a different treatment, since January 28, after a preliminary period of training. In each category of birds we had a control group receiving only the basal diet. In young roosters we had two more groups that were fed thyroprotein at three different levels: Group B received thyroprotein as 0.04% of the ra- tion throughout the period of experiment; Group 0 received two different levels during the experimental period: from January 28 until Feb- ruary 29 it received 0.08% of the ration; in the last two weeks it received twice this dose, that is, 0.16% of the ration. In the old roosters we had a control group (group D), receiving the basal mash, and a group (group E) -45- receiving thyroprotein as 0.04% of the feed. The results obtained in this experiment showed that thyroprotein when given as 0.04% of the feed is able to improve spermatogenesis in both young and old roosters. We obtained an increase in semen volume, sperm concen- tration and consequently, in the total number of sperm. The results from young roosters are more consistent than in the older ones. Higher levels of thyroprotein seemed to have a depressing effect on spermatogenesis, as can be seen in the data obtained from group C of the young roosters. We observed a decrease in semen volume, sperm concen- tration, and consequently, in the total number of sperm in this group. -46... 1. CONCLUSIONS Therprotein when given as 0.04 per cent of the ration produced an increase in spermatogenesis of young roosters. As compared to control birds both semen volume and sperm concentration increased to some extent with the result that the total Sperm per ejaculate increased rather markedly. In old- er roosters similar but less pronounced trends were observed. Thyroprotein when given at the higher levels of 0.08 and 0.16 per cent of the ration, depressed semen production and the total number of sperm decreased markedly o -47- Ii EF Eh‘ENC ES Aron, N. and Benoit, J., 1934. Comptes rendus Soc. Biol. 117:218. Astwood, E. B., 1943. Treatment of hyperthyroidism with thiourea and thiouracil. J. A. M. A., 22:78. Baumann, 8., 1896. Ueber das normale vorkommen von jod im thierkorper. Z. Physiol. Chem., 21:319. Benoit, J. and Aron, N., 1934. Comptes rendus. Soc. Biol., 116:221. Benoit, J. 1936. Role de la thyroide dans la gonado- stimulation par la lumiere artificielle chez le canard domestique. Soc. Biol., 123:243. Benoit, J., 1937-a. Thyroide et croissance testiculaire chez le canard domestique. 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