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then stained in haematoxylin and eosin for histopathological studies (Kiernan, 2008). For statistical evaluation of significance, 100 seminiferous tubules (‘a’ and ‘c’), per group were assessed and X 2 (Chi Square) test was conducted using the formula as per the two by two table. Results In the present study the testes of cadmium and curcumin treated animals were assessed using the following parameters: Changes in morphology, testicular pathology, and cytostatic and cytotoxic changes in the germ and Leydig cells. The testes of control group mice administered only vehicle showed normal pathology with distinct seminiferous tubules undergoing different stages of spermatogenesis. The interstitium was compact with distinct Leydig cells (Fig.1). Spermatogonial mother cells, primary and secondary spermatocytes, maturing spermatids and spermatozoons embedded in Sertoli cells were clearly visible (Fig.2). Histopathological evaluation of only curcumin treated mice testes showed normal structures similar to control group. Testes of the cadmium treated animals exhibited hemorrhage of its vasculature, but the overall shape was not altered. Seminiferous tubules appeared to lose their typical spherical shape and tunica propria manifested thickening and was irregularly and randomly broken at places. Leydig cells were entrapped in the degenerated interstitium (Fig. 3). There was general derangement of morphology of spermatogonia, Curcumin protects against cadmium chloride 33 where ‘b’ the Observed Frequency for category ‘a’ ‘d’ is the Expected Frequency in the corresponding category ‘c’ This experimental study was done after taking approval from the Institutional Animal Ethics Committee (No.Cs\Res\07\759). a b a+b c d c+d a+c b+d N ; X 2 = N (ad-bc) 2 (a+c ) ( b+d) ( a+b) (c+d) spermatocytes and differentiating spermatids in the seminiferous tubules (Fig. 4). In most of the peripheral seminiferous tubules, the germ cells were seen to break free end appeared in the lumen as puffs (Fig. 5). The cell types which appeared to be most affected were spermatocytes and spermatids. Tubular lumens were filled with degenerated germ cells and multinucleated spermatid aggregates (Fig. 3). Vacuolization of the seminiferous epithelium was also observed (Fig. 6). The results show that a single dose of cadmium causes a sudden increase in testicular damage, apparently overpowering this tissue’s natural defenses. Also, most of the seminiferous tubules of testes in this group showed complete absence of primary spermatocytes, secondary spermatocytes, spermatids and spermatozoa and loss of spermatogenesis process (Fig. 6) in comparison with normal structure of seminiferous tubules in control mice. The seminiferous tubules of experimental group animals pre-treated with curcumin for 15 days and cadmium chloride on 16 th day showed very slight histopathological damage in the peripheral tubules and the inner tubules appeared to be normal showing all stages of spermatogenesis viz. spermatoginal mother cells, primary and secondary spermatocytes, maturing spermatids and spermatozoons embedded in Sertoli cells (Fig.7 and 8).
34 Preeti SINGH et al. Figure 1. 1) Photomicrograph of testis of control group mice administered only vehicle. Normal seminiferous tubules (ST) and Leydig cell (LC) are clearly visible (20X). 2) Testis of control group mice administered only vehicle. Seminiferous tubule with distinct spermatogonial mother cells (SMC), primary spermatocyte (PS), secondary spermatocytes (SS), maturing spermatids (MS) and spermatozoa (S) are seen (40X). 3) Testis of cadmium chloride (50mg/kg) treated group mice. Degenerate seminiferous tubules(ST) are clearly visible (20x). 4) Testis of cadmium chloride(50mg/kg) treated group mice. Degenerated germ cells in lumen of seminiferous tubules are clearly evident (40x). 5) Testis of cadmium chloride(50mg/kg) treated mice showing exfoliated germ cells(GC) in tubular lumen (100x). 6) Testis of cadmium chloride (50mg/kg) treated mice. Seminiferous tubules are left with only vacuolised spermatogonial mother cells (SMC) and Sertoli cell (SC) (100x). 7) Testis of group pre-treated with curcumin(10mg/kg for 15 days) and then administered cadmium chloride(50mg/kg). Seminiferous tubules (ST) showing stages of spermatogenesis and pyknotic nuclei in Leydig cells (LC) are clearly evident (40x). 8) Testis of group pre-treated with curcumin (10mg/kg for 15 days) and then administered cadmium chloride(50mg/kg). Maturing spermatids (MS) and adhered germ cells are visible (100x)
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