1 - دانش آموخته مامایی و بیماریهای تولید مثل دامپزشکی، دانشکدة علوم تخصصی دامپزشکی، دانشگاه آزاد اسلامی واحد علوم و تحقیقات تهران، تهران، ایران
2 - گروه علوم درمانگاهی، دانشکدة دامپزشکی، دانشگاه تهران، تهران، ایران
3 - گروه علوم درمانگاهی، دانشکدة دامپزشکی، دانشگاه تهران، تهران، ایران
4 - گروه علوم درمانگاهی، دانشکدة دامپزشکی، دانشگاه آزاد اسلامی واحد تبریز، تبریز، ایران
5 - گروه علوم درمانگاهی، دانشکدة دامپزشکی، دانشگاه شهرکرد، شهرکرد، ایران
تاریخ دریافت : 1387/08/09
تاریخ پذیرش : 1387/10/30
تاریخ انتشار : 1387/09/01
هدف از این مطالعه بررسی اثرات سه غلظت اسید سیتریک بر الگوی حرکتی اسپرمهای اپیدیدیمی گاو بوده است. برای اینکار 50 جفت بیضة گاو از کشتارگاه صنعتی ارومیه بلافاصله بعد از کشتار جمعآوری و در کنار یخ 5 درجه سانتیگراد به آزمایشگاه انتقال داده شد. اسپرمهای ناحیة دم اپیدیدیم با چند برش در ناحیة بدون رگ آن جمعآوری و به محیط Hams F10 با 10% سرم گوسالة جنینی و دمای 37 درجة سانتیگراد انتقال و پس از 15 دقیقه انکوباسیون در انکوباتور CO2 شمارش گردید و رقتهای 50 میلیون اسپرم در میلیلیتر آماده و در محدودة pH نرمال اسپرم (7/6 الی 4/7) سه غلظت اسید سیتریک 1/0، 2/0 و 3/0 نرمال (غلظت 1 نرمال اسید سیتریک، 7 میلیگرم در 1 میلیلیتر مایع منی گاو است) تهیّه شد و به اپندورفهای حاوی اسپرم اضافه گردید و در دقایق 15،30، 45، 60، 90، 120، 180، 240 و360 الگوی حرکتی اسپرمهای اپیدیدیمی با روش کاسا ارزیابی شد. دادههابا نرمافزارSPSS ویرایش 15 و آزمون آنالیز واریانس یکطرفه، آنالیز گردید. نتایج نشاندهندة اختلاف معنیدار شاخصههای مختلف الگوی حرکت اسپرمی (سرعت خط منحنی، سرعت خط مستقیم ، خطی بودن، درصد حرکت رو به جلو) خصوصاً در غلظت 3/0 نرمال در مقایسه با شاهد بود (05/0p<).
چکیده انگلیسی:
The aim of this study was to investigate the effect of three concentration of citric acid on motility patterns of bovine epididymal sperms. For this purpose, 50 pairs of bovine testicles were collected immediately after slaughter form urmia abattoir and transferred to the laboratory alongside 5oc ice pack. Epididymal tail sperms were collected with a few incisions in vascular areas and transferred to hams f10 milieu with 10% fetal calf serum and counted after 15 minutes of incubation at 37oc in Co2 incubator. Concentrations of 50 million sperms per ml were proposal and in the normal sperm pH rang of 6.7-7.4, 0.1, 0.2 and 0.3 normal concentration of citric acid were added to sperm continuity micro tubes (normal concentration of acid equals 7 mg/ml of bovine serum) and at 15, 30, 45, 60, 90, 120, 180, 240 and 360 minutes the motility patterns of epididymal sperms were evaluated using the computer assisted sperms analyzing (CASA) method. Data were analyzed with one-way ANOVA using the SPSS 15 software. The results indicated significant differences in various indices of sperm motility patterns (Curvilinear Velocity, Straight-line Velocity, Average Path Velocity, Mean Angel Degree, Amplitude of Lateral Head Displacement, Beat-Cross Frequency, Linearity, Wobble) particularly at 0.3 normal concentration of citric acid compared with the control.
منابع و مأخذ:
Amann, R.P. (1989): Can fertility potential of a seminal sample be predicted accurately? Journal of Andrology, 10: 89-98.
Amann, R.P., Seidel, G.E. and Mortimer, R.G. (2000): Fertilizing potential in vitro of semen from young beef bulls containing a high or low percentage of sperm with a proximal droplet. Theriogenology, 54: 515-1499.
Auger, J., Leonce, S., Jouannet, P. and Ronot, X. (1993): Flow cytometric sorting of living, highly motile human spermatozoa based on evaluation of their mitochondrial activity. J. Histochem. Cytochem., 4 1: 1247-1251.
Cha, S.Y., Tsoi, W.L., Leung, J., Ng, V., Lo, T. and Wang, C. (1990): The accuracy of sperm concentration determination by the automated Cell Soft semen analyzer before and after discontinuous percoll gradient centrifugation. Andrology, 2: 55-61.
Cox, J.F., Alfaro, V., Montenegro, H. and Rodriguez, M. (2006): Computer-assisted analysis of sperm motion in goats and its relationship with sperm migration in cervical mucus. Theriogenology, 66: 860-867.
Elzanaty, S., Johan, M. and Aleksander, G. (2005): Duration of sexual abstinence: epididymal and accessory sex gland secretions and their relationship to sperm motility. Human Reproduction, 20: 221-225.
Gillan, L., et al. (2007): Assessment of in vitro sperm characteristics in relation to fertility in dairy bulls, Anim. Reprod. Sci., doi: 10.1016/j.anireprosci.2006.12.010., In press.
Goovaerts, G.G., Hoflack, A., Van Soom, J., Dewulf, M., Nichi, A. and de Kruif, P.E.J. (2006): Evaluation of epididymal semen quality using the Hamilton–Thorne analyzer indicates variation between the two-caudae epididymides of the same bull. Theriogenology, 66: 323-330.
Harayama, H. and Kato, S. (1992): Changes in motility and morphology of spermatozoa during their transit through the epididymis in Meishan boars at various ages. Anim. Sci. Technol. (Jpn), 63: 462-467.
Hishinuma, M., Suzuki, K. and Sekine, J. (2003): Recovery & cryopreservation of sika deer (Cervus nippon) spermatozoa from epididymides stored at 4 degrees. Theriogenology, 59: 813-820.
Herolda, J.E. and Aurichb, D.G. (2004): Epididymal sperm from African buffalo can be frozen successfully with AndoMed1 & Triladyl TM but the addition of bovine seminal plasma is detrimental. Theriogenology, 61: 715-724.
Holt, W., Watson, P., Curry, M. and Holt, C. (1994): Reproducibility of computer-aided semen analysis: Comparison of five different systems used in a practical workshop. Fertility and Sterility, 62: 1277-1282.
Ikeda, H., Kikuchi, K., Noguchi, J., Takeda, H., Shimada, A., Mizokami, T., et al. (2002): Effect of preincubation of cryopreserved porcine epididymal sperm. Theriogenology, 57: 1309-1318.
Janulis, L., Hess, RA., Bunick, D., Nitta, H., Janssen, S., Asawa, Y. and Bahr, J.M. (1996): Mouse epididymal sperm contain active P450 aromatase, which decreases as sperm traverse from the epididymis. Journal of Andrology, 17: 111-116.
Kaabi, M., Paz, P., Alvarez, M., Anel, E., Boixo, G.C., Rouissi, H., Herraez, P. and Anel, L. (2003): Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem. Theriogenology, 60: 1249-1259.
Kathiravan, P., et al. (2007): Computer automated motion analysis of crossbred bull spermatozoa and its relationship with in vitro fertility in zona-free hamster oocytes, Anim. Reprod. Sci., doi: 10.1016/j.anireprosci.2007.01.002., In press.
Kato, S., Toshitaka S.A., Hiroshi, H. and Yasuyuki, K. (1996): Timing of shedding and disintegration of cytoplasmic droplets from boar and goat spermatozoa. Journal of Reproduction and Development, 42: 237-241.
Kato, S., Yasui, T. and Kanda, S. (1987): Changes in motility and morphology of goat spermatozoa during their transit through the epididymis. Mem Grad School Sci. & Technol, Kobe University, 5B: 35–42.
Martinez-Pastor, C., Guerra, M., Kaabi, A.R., Diaz, E., Anel, P., Herraez, P. and de Paz, L. (2005): Decay of sperm obtained from epididymis of wild ruminants depending on postmortem time. Theriogenology, 63: 24-40.
Margo, L., Macpherson (2001): How to Evaluate Semen in the Field. Vol: 47, AAEP (American Association of Equine Practitioners) Proceedings, pp: 413-414.
Mortimer, S.T. and Mortimer, D. (1990): Kinematics of human spermatozoa incubated under capacitating conditions. J. Androl., 11: 195-203.
Naomi, K., Ena, N., Akira, K., Chikashi, T. and Masao, S. (2001): Freezing epididymal spermatozoa of the Japanese serow (Capricornis crispus) in liquid nitrogen. Journal of Reproduction and Development, 47(6): 359-363.
Pe´rez, S., Aguado, M., Ayllon, E., Garrido, D., Montoro, V. and Garde, J. (1995): Live birth of hybrid (O. musimon-O.aries) lambs following intrauterine insemination in domestic sheep with mouflon semen obtained 40 hours postmortem. Theriogenology, 43: 218.
Rao A.R., Bane, A. and Gustafsson, B.K. (1980): Changes in the morphology of spermatozoa during their passage through the genital tract in dairy bulls with normal and impaired spermatogenesis. Theriogenology, 14: 1-12.
Verstegen, J., Iguer-Ouada, M. and Onclin, K., (2002): Computer assisted semen analyzers in andrology research and veterinary practice. Theriogenology, 57: 149-179
Youngquist, et al. (2007): Current Therapy in Large Animal Theriogenology. Saunders, London, Chapter 30, pp: 221-237.