VolumeInternational II Number Journal 2 2011 for [1-7] Environmen tal Rehabilitation and Conservation Volume[ISSN 0975 II No. - 6272] 2 2011 [1-7] Charde [ISSN et al.0975 - 6272]

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain , coromandelica.

Pravin Charde1, Jyoti Ramteke1 and Suresh Zade2

Received: January 2, 2011 ⏐ Accepted: July 3, 2011 ⏐ Online: October 20, 2011

Abstract Introduction are almost tailless patridge like Adult rain quail are sexually diamorphic. The popularly known as “Batter”, belonging to the aim of this study was to determine the effect class Aves and family . Batter is of testosterone propionate and hydroxyl a good table bird known for its delicacy since progesterone caproate on gonads and some olden days. They are used as food before other endocrine glands of male and female chicken was domesticated. Meat is rich in rain quail. 15 male and 15 female rain quails vitamins, amino acids, unsaturated fatty acid were treated with testosterone and etc., which are very vital for health of human progesterone daily for 7 and 15 days being. Quail meat therefore can be respectively. The study reveals that after the recommended to be included in diet of treatment of testosterone, the spermatogonia children, pregnant mother and convalescent were detached and show hypertrophy as well patient for speedy recovery. as the germ cells were enlarged and vacuolated every degenerates, thyroid and Perusal of literature on quail reproduction reveals that, little is known about these adrenal shows hypertrophy. aspects in exotic and tropical species. The Keywords: Testosterone Propionate ⏐ reason is that more critical experiments have Hydroxyl Progesterone Caproate⏐Sexual not been successfully undertaken in Indian Dimorphism ⏐ Gonads ⏐ Rain Quail. quail species. Studies on these species are confined mainly to the changes in the

reproductive organs and rarely endocrine glands during different phases of reproductive For Correspondence: cycle. No experimental studies have been 1Principal, Sevadal Mahila Mahavidyalaya, Nagpur- made in these species endocrine glands in 440009 (M.S.) reproduction. Fundamental knowledge of 2 Professor, P.G.T.D. Zoology, Rashtrasant Tukadoji reproductive process and the endocrine Maharaj Nagpur University, Nagpur (M.S.) mechanism that control process is therefore,

Email: [email protected] invisible for profitable management and production of these in large scale. The paucity of information on reproduction

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain quail, Coturnix coromandelica. 1 Volume II Number 2 2011 [1-7] [ISSN 0975 - 6272] Charde et al.

endocrinology of Indian quail motivated me were acclimatized in laboratory for at least 2 to undertake this investigation. to 3 weeks.

The present study was undertaken on rain The males are divided into four groups quail, Coturnix coromandelica. This bird was of 15 each. The ground II and IV are selected for this work because it offers the given intramuscular injection of testosterone advantages of endocrinological works. The propionate 0.15 mg/kg/day for 7 and 15 aim and objective this investigation is to study consecutive days respectively. The I and III the histopathological effect of testosterone group receive equivalent volume of propionate and hydroxy progesterone physiological saline for the same period. For caproate on gonads and other endocrine another experiment 40 female birds were glands and to investigate the mechanism of divided into four groups of 10 birds each. The action of this steroids in both sexes of this group II and IV were given intramuscular Rain quail. injection of hydroxy progesterone caproate

In the present study testosterone propionate 0.15 mg/kg/day consecutive for 7 and 15 days and hydroxyl progesterone caproate obtain respectively. The I group and II were used as from German Remedies Limited, Bombay a control and treated with equivalent volume under trademark of Schering AG, West- of physiological saline for the same period. German. Birds of experimental and control groups were weighed before and at the end of the MATERIAL AND METHODS experiments and maintained under same husbandry condition. At the end of each The Indian rain quail C. coromandelica has experimental animals were sacrificed. For been selected for present investigation. Adult histopathological studies reproductive organs quail are sexually dimorphic. Reproductive and endocrine glands were dissected, weighed behaviour is sexually differentiated in quail. and fixed in various fixatives. All endocrine The adult rain quail is about 6½ to 7 inches in glands were fixed in formal sublimate for 24 size (without tail) and body weight is 85 to 90 Hrs. and other tissue were fixed in Bouin’s gms. Temperature, light, floor space, fluid. The tissue were dehydrated in various humidity and feed are most important factors grades of alcohol cleared in xylene and after for good housing and maintenance of quails. embedded in paraffin blocks were prepared. The maximum daily amount of feed given Paraffin sections were cut at 5 to 6 µm was equivalent to 20 to 25% of body weight. thickness. Sections were stained with To identify individual quails from each haematoxyline and eosin. groups, they were marked with numbered aluminium strips using bands. HISTOMORPHOLOGY The rain quail utilized in this study were Testis – The testis consists of compact collected from various places in Nagpur convoluted seminiferous tubules which are district and Amravati district of Maharashtra lined by single or double layer of germinal State. The birds were trap over a period of cells. The tubules are composed of primary four months during October to January and spermatogonia, secondary spermatogonia and consisted of 60 adult males and 40 adult sectoli cells. Further stages of females weighing 70 to 90 gms. Before spermatogenesis such as spermatocytes, commencement of the experiments all birds spermatids and spermatozoa are not observed. Similarly, the sertoli cells are inconspicuous

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain quail, Coturnix coromandelica. 2 Volume II Number 2 2011 [1-7] [ISSN 0975 - 6272] Charde et al.

and regressed. Since, the control males are sacrificed in the month of January which is sexually inactive period in rain quails, the testis have showed regression. In the interfollicular region blood capillaries, connective tissue and regressed Leydig’s cells are present.

Figure 2: Photomicrograph of control ovary showing primary follicles and germinal follicles with theca externa, theca interna and interstitial cells.

Adrenal gland of rain quail is paired, triangular in shape and yellow or orange in colour. It locates anterior and medial to the cephalic lobe of kidney and posterior to the lungs. It is surrounded by thick capsule which Figure 1: Photomicrograph of control testis is made up of connective tissue. showing convoluted seminiferous tubules with germ cells, sertoli cells and interstitial Pineal gland is present on the dorsal side of cells. the brain in triangular space between Ovary: cerebellum and cerebral hemisphere. Pineal In the rain quail single ovary and oviduct is grain in rain quail is creamisa brown, club- present which is situated on the left side of the shaped conical structure. Pineal gland is cephalic end of kidneys and attached the body composed of lobules and each lobule consists wall by ligament. In rain quail, the ovary of follicles which are separated by contains several primary follicles and blood intralobular septa.

haemorrhage. Developing follicles are Thyroid gland of the male and female rain composed of germinal vesicle granulose cells, quail is composed of two separate lobes theca interna and theca externa. Some degree which are reddish in colour, situated on either of atresia is observed at primary follicular side of the trachea in between jugular and stage. Rarely degeneration is visible in large carotid blood vessels. The normal thyroid follicles. gland is encapsulated and composed of well organized compact follicles.

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain quail, Coturnix coromandelica. 3 Volume II Number 2 2011 [1-7] [ISSN 0975 - 6272] Charde et al.

Parathyroid gland in the rain quail is paired, more pronounced in the fifteen days treated well vascularised and closely attached to the birds. posterior part of the thyroid gland. Adrenal: 7 days after treatment: No significant changes RESULTS AND DISCUSSION were found in the vasculature of the gland. No Pineal: changes were evident in Type II cells while 7 days after treatment: No significant Type I cells showed slight hypertrophy and changes were observed in the vasculature of degradation. pineal gland. Lumen of the follicle was 15 days after treatment: More significant slightly obliterated. Type-I i.e. polygonal cells changes were seen in the internal cells. The shows regression whereas Type-II i.e. tall internal cells shows hypertrophy and were cells shows hypertrophy and degranulation. degranulated and vacuolated. No significant changes were observed in Type II and showed 15 days after treatment: Showed more little hypertrophy as compared to control. pronounced hypertrophy of Type-II i.e. tall

columnar cells as compared to 7 days of Testis: testosterone propionate. The cell debris in 7 days after treatment: lumen was evident. Type-I i.e. polygonal cells showed more regression.

Thyroids: 7 days after treatment: Testosterone propionate treatment resulted in hypertrophy of the follicular epithelial cells and thickness of follicular wall. It induced hypertrophy and hyperplasia in small follicles. Large follicles showed little regressive effect.

15 days after treatment: Induced changes in thyroid follicles. Diameter of follicles were increased. The gland contained several empty follicles. The colloid contained few vacuoles at the peripheral region. The reduction in the amount of colloid was more evident.

Parathyroid: Figure 3: Photomicrograph of testis 7 days 7 days after treatment: Testosterone treatment of testosterone propionate showing propionate treatment induced hypertrophy and detached spermatogonia in the lumen.

vacuolization in the cells of parathyroid cells The germinal epithelium was not affected of parathyroid gland. Small circular vacuoles after treatment of testosterone propionate. appeared adjacent to the nucleus. Debris formation in the lumen of the

15 days after treatment: The above mentioned seminiferous tubules was seen. Regressive effect of testosterone propionate were same, changes were observed in peripheral rather more pronounced were same, rather seminiferous tubules. Primary spermatogonia and secondary spermatogonia were detached

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain quail, Coturnix coromandelica. 4 Volume II Number 2 2011 [1-7] [ISSN 0975 - 6272] Charde et al.

and seen accumulated in the lumen of viscous. The large follicles shows regressive seminiferous tubules. In some tubules lumen changes. disappeared because of hypertrophy of germ 7 days after treatment: cells. No changes were observed in the vasculature of gland.

15 days treatment:

Figure 5: Photomicrograph of Ovary 7 days treatment of hydroxyl progesterone caproate showing primary follicles and formation of space between ovarian stroma and blood haemorrhage

15 days after treatment:

Figure 4: Photomicrograph of testis 15 days treatment of testosterone propionate showing completely obligated lumen, hypertrophy of sertoli cells and degenerative effect on Leydig’s cells (Interstitial cells) Figure 6: Photomicrograph of Ovary 15 days treatment of hydroxyl progesterone caproate In most of the tubules germ cells detached showing greatly regressed germinal vesicle, and accumulated as debris in the lumen. Germ degenerating large follicles and disturbed cells were enlarged and vacuolated. Lumen of interstitials cells. seminiferous tubules disappear enlarged size of the primary spermatogonia and secondary The space between blood haemorrhage and spermatogona showed hypertrophy. Reydig’s ovarian stroma is more pronounced. Follicular cells were degenerated while sertoli cells cytoplasm and germinal vesicle shows showed hypertrophy. Blood vessels because regressive changes. There was no significant dilated. effect on the theca interna and theca externa, but these two layers started to separate from Ovary each other in large follicles. In stroma interstitial cells shows more degenerative The atretic primary follicles and formation of changes. space between the haemorrhage and ovarian

stroma changes. Haemorphage become more

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain quail, Coturnix coromandelica. 5 Volume II Number 2 2011 [1-7] [ISSN 0975 - 6272] Charde et al.

CONCLUSION: Bently, P. J. (1976). Comparative invertebrate From the present study it has been concluded endocrinology, Cambridge University that the doses of testosterone propionate Press, Cambridge, London, New stimulates the spermatogenesis in male rain York. quail. Similarly the treatment of hydroxyl Bhatia, A.J. and Wade, G.N.(1989). progesterone caproate in female stimulates the Progesterone can either increase or secretion of FSH of pituitary and decrease weight gain and adiposity in development of ovarian follicles. ovariectomized Syrain Hamsters Physiology and Behaviour, 46:273- 278. References C. Berg, L. Holni, I. Brandt and B. Abalain, J. H., Amet, Y. Dahiel, J. Y. and Brunstrom, (2001) Anatomical and Floch, H. H. (1983). Testosterone histological changes in the oviduct of stimulation of DNA- Dependant DNA coturnix japonica, after polymerase activities in the cloacal embryonic exposure to and uropygial glands of the male quail ethynyloesstradiol. Reproduction 121, (coturnix coturnix japonica). Gen. 155-165. Comp. Endocrinology. 52, 164-171. Charde, P. N. (1998). Influence of Exogenesis Alison, M. R. and Wright, N.A. Testosterone Pharmacological Compounds on 5 alpha reductase activity as related to Reproductive Biology of Coturnix proliferative status in mouse accessory coromandelica. Thesis submitted for sex glands: J. endocrinology; 81;83- Ph.D. Degree Nagpur University, 92, 1979. Nagpur. Ambadkar, P. M. and Chauhan, B. C. (1980). Follet, B. K. (1984a). In Marshal’s Observations on influences of Physiology of Reproduction. Vol. 1: pinealectomy on certain aspects of Reproductive Cycles of Vertebrates testicular functions in Indian house (G.E. Lamming Ed.), pp.283-350. crow (Crovus splendens). Pavo 18, 39- Fuladi, R. S. (1992). Histopathological and 52. biochemical effects of some drugs on Arneja, D.V., George, G.C., Mehta, S.N. gonads and endocrine glands of grey Dixit, V.P. and Razdan, M. N. (1984). quail, coturnix coturnix. Thesis Plasma testosterone concentration in submitted for Ph.D. Degree, Nagpur relation of thyroid status in Japanese University, Nagpur. quail. Indian J. Poultry Science, 19, Gorbman, A. and Bern, H. A. (1962). A 207-209. textbook of comparative Arneja, D.V., Sharma, D. W. and Razdan, endocrinology, John Wikey and Sons, M.N. (1985). Histomorphology of New York. parathyroids in white perkin drakes. Hervey, E. and Hervey, G. R. (1967). The Ind. Journal Poultry Sci. Vol.20, 58- effects of progesterone on body 60. weight and composition in the rat.

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain quail, Coturnix coromandelica. 6 Volume II Number 2 2011 [1-7] [ISSN 0975 - 6272] Charde et al.

Journal of Endocrinology. 37: 361- Distribution and effects of testosterone 384. on aromatase mRNA in the quail M.A.Ottinger, S. Pitts and M.A. Abdelnabi forebrain. A non-radioactive in situ (2001). Steroid hormones during hybridization study. Journal of embryonic development in Japanese Chemical Neuroanatomy 14, 103-115. quail; plasma gonadal, and adrenal levels. Paultry Science 8; 795-799.

N. Aste, G.C. Panzica, C. Viglietti. Panzica, N. Harada, J. Balthazar (1998).

Histopathological effect of testosterone propionate on male and hydroxyl progesterone caproate on female rain quail, Coturnix coromandelica. 7