Effect of Different Levels of Egg Yolk on Cryopreservation of Black Bengal Buck Semen in Tris Egg yolk Citrate Fructose Glycerol Extender
محورهای موضوعی : Camelال. گویجن سینق 1 , ک. رای 2 , ب. سرکار 3
1 - Department Veterinary Gynaecology, Directorate of Veterinary and A.H. Services, Manipur, Sanjenthong, India
2 - Department of Veterinary Gynaecology and Obstetrics, Faculty of Veterinary and Animal Science, WBUAFS, 37, K.B. Sarani, Kolkata, 700037, West Bengal, India
3 - Department of Veterinary Gynaecology and Obstetrics, Faculty of Veterinary and Animal Science, WBUAFS, 37, K.B. Sarani, Kolkata, 700037, West Bengal, India
کلید واژه: buck, cryopreservation, glycerol, semen extender,
چکیده مقاله :
This study was conducted to assess the physical and morphological characteristics of fresh, pre-freeze and post-thaw black Bengal buck (Capra hircus) semen processed in Tris-egg yolk-citrate-fructose-glycerol (TEYCFG) extender containing 4 different concentrations (2.5, 5, 7.5 and 10%) of egg-yolk. Ejaculates were collected once a week for eight weeks from six mature bucks (48 ejaculates) using an artificial vagina. It was found that a general decrease in values of pre-freeze and post-thaw semen parameters such as progressive sperm motility (%), live sperm (%), acrosomal integrity (%) and positive sperm (%) in the hypo-osmotic swelling (HOS) test, but an over-all increase in abnormal spermatozoa compared to fresh semen for extenders containing different concentrations of egg-yolk. Best values for all the semen parameters were obtained using extender with 2.5% egg-yolk. Of all egg-yolk concentrations, the 10% egg-yolk supplement caused the highest percentage of abnormal sperm, which was significant (P<0.05), compared to 2.5% and 5% egg-yolk containing extenders, but non-significantly (P>0.05) different compared with 7.5% egg-yolk containing extender. Only progressive motility was significantly different (P<0.05) within extenders containing 2.5%, 5%, 7.5% and 10% egg-yolk. We conclude that Black Bengal buck semen can be cryopreserved effectively with tris-egg yolk-citrate-fructose-glycerol extender containing 2.5% egg yolk (V/V).
این مطالعه برای ارزیابی خصوصیات فیزیکی و مرفولوژیکی منی تازه و منی پیش از انجماد و یخ گشایی شده بز سیاه بنگال انجام شد. نمونهها در رقیقکننده تریس-زرده تخم مرغ-سیترات-فروکتوز-گلیسرول دارای 4 غلظت متفاوت (5/2، 5، 5/7 و 10 درصد) از زرده تخم مرغ عملآوری شدند. انزالها (48 انزال) به صورت هفتگی و به مدت 8 هفته از 6 بز بالغ با استفاده از واژن مصنوعی جمعآوری شد. یک کاهش عمومی در مقادیر فراسنجههای پیش و پس از انجماد از قبیل حرکت پیشرونده اسپرم (درصد)، اسپرم زنده (درصد)، یکپارچگی اکروزوم (درصد) و اسپرم مثبت (درصد) در آزمون HOS، ولی یک افزایش کلی در اسپرمهای غیر طبیعی در مقایسه با اسپرم تازه برای رقیقکنندههای حاوی غلظتهای متفاوت زرده تخم مرغ مشاهده شد. بهترین مقادیر برای تمامی فراسنجههای منی با استفاده از رقیقکننده حاوی 5/2 درصد زرده تخم مرغ به دست آمد. از بین غلظتهای زرده تخم مرغ، غلظت 10 درصد بالاترین اسپرم غیر طبیعی را سبب شد، که در مقایسه با غلظتهای 5/2 و 5 درصد معنیدار (05/0P<)، ولی در مقایسه با غلظت 5/7 درصد غیر معنیدار بود (05/0P>). حرکت پیشرونده اسپرم تنها فراسنجهای بود که به طور معنیداری (05/0P<) بین غلظتهای متفاوت رقیق کننده (5/2، 5، 5/7 و 10 درصد) متفاوت بود. نتیجهگیری اینکه منی بز سیاه بنگال میتواند به طور مؤثری با استفاده از رقیقکننده تریس-زرده تخم مرغ-فروکتوز-گلیسرول حاوی 5/2 درصد زرده تخم مرغ (v/v) در شرایط انجماد نگهداری شود.
Aboagla E.M.E. and Terada T. (2004). Effects of egg yolk during the freezing step of cryopreservation on the viability of goat spermatozoa. Theriogenology. 62, 1160.
Aires V.A., Hinsch K.D., Schloesser F.M., Bogner K., Schloesser S.M. and Hinsch E. (2003). In vitro and in vivo comparison of egg yolk based and soya bean extenders for cryopreservation of bovine semen. Theriogenol. 60, 269-279.
Aitken R.J., Clarkson J.S. and Fishel S. (1989). Generation of reactive oxygen species, lipid peroxidation, and human sperm function. Biol. Reprod. 41, 183-197.
Apu A.S., Khandoker M.A.M.Y., Hussain S.S., Fakruzzaman M. and Notter D.R. (2012). A comparative study of fresh and frozen-thawed quality in relation to fertility of black Bengal goats. Iranian J. Appl. Anim. Sci. 2(2), 157-161.
Ashmawy T.A.M., Sallam A.A., El-Khalek A.E.A., El-Saidy B.E. and Gabr M.G. (2010). Recovery and fertilization rates of goat spermatozoa as affected by different levels of egg yolk, dilution rates, freezing method and months of the year. Egypt J. Sheep. Goat. Sci. 5(1), 283-293.
Bailey J.L., Bilaodeau J.F. and Cormier N. (2000). Semen cryopreservation in domestic animals: a damaging capacitating phenomenon. J. Androl. 20, 1-7.
Barrea-Compean M.H., Purdy P.H., Dzakurna J.M., Newton G.R. and Nuti L.C. (2005). Cholesterol-loaded cyctodextrin improves post-thaw goat sperm motility. J. Anim. Sci. 83(1), 153-153.
Batista M., Nino T., Alamo D., Castro N., Santana M., Gonzalez F., Cabrera F. and Gracia A. (2009). Successful artificial insemination using semen frozen and stored by an ultrafreezer in the Majorera goat breed. Theriogenology. 71, 1307-1315.
Bispo C.A.S., Pugliesi G., Galvao P., Rodrigues M.T., Ker P.G., Filgueiras B. and Carvalho G.R. (2011). Effect of low and high egg yolk concentrations in the semen extender for goat semen cryopreservation. Small Rumin. Res. 100, 54-58.
Bousseau S., Brillard J.P., Guienne M., Guerin B., Camus A. and Lechat M. (1998). Comparison of bacteriological qualities of various egg yolk sources and then in vitro fertilizing potential of bovine semen frozen in egg yolk or lecithin based diluents. Theriogenology. 50, 669-706.
Cabrera F., Gonzalez F., Batista M., Calero P., Medrano A. and Gracia A. (2005). The effect of removal of seminal plasma, egg yolk level and season on sperm freeze ability of canary buck (Capra hircus). Reprod. Domest. Anim. 40, 191-195.
Curry M.R. and Watson P.F. (1994). Osmotic effects on ram and human sperm membranes in relation to thawing injury. Cryobiology. 31, 39-46.
Curry M.R., Kleinhans F.W. and Watson P.F. (2000). Measurement of the water permeability of the membranes of boar, ram, and rabbit spermatozoa using concentration-dependent self-quenching of an entrapped fluorophore. Cryobiology. 41, 167-173.
Deleeuw F.E., Chen H.C., Colenbrander B. and Verkleij A.J. (1990). Cold-induced ultrastructural changes in bull and boar sperm plasma membranes. Cryobiology. 27, 171-183.
EI-Maghraby M.M. (2007). Physiological studies on reproduction in goats. Ph D. Thesis. MonsouraUniv., Monsoura, Egypt.
Foulkes J.A. (1977). The separation of lipoprotein from egg yolk and their effect on the motility and integrity of bovine spermatozoa. J. Reprod. Fertil. 49, 277-284.
Halliwell B. (1991). Reactive oxygen species in living systems: source, biochemistry and role in human disease. Am. J. Med. 91, 14-22.
Hancock J.L. (1951). A staining technique for the study of temperature-shock in semen. Nature. 167, 323-324.
Hancock L. (1952). The morphology of bull spermatozoa. Br. J. Exp. Biol. 29, 445-453.
Iritani A. and Nishikawa Y. (1972). Studies on the egg yolk coagulation enzyme (Phopholipase) in goat semen. IX. Enzyme concentration in the semen collected from the Cowper’s gland removed goat. Mem. Coll. Agric. Kyoto Univ. 10, 44.
Jeyendran R.S., Vander Ven H.H., Perez-Pelaez M., Crabo B.G. and Zaneveld L.J.D. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil. 70, 219-228.
Kumar S. and Das G.K. (2005). Frozen sperm quality with reference to reactive oxygen species: a review. Indian J. Anim. Sci. 75, 874-884.
Maxwell W.M.C. and Salamon S. (1993). Liquid storage of ram semen. Reprod. Fertil. Dev. 5, 613-638.
Maxwell W.M.C. and Watson P.F. (1996). Recent progress in the preservation of ram semen. Anim. Reprod. Sci. 42, 55-65.
Medeiros C.M., Forell F., Oliveira A.T. and Rodrigues J.L. (2002). Current status of sperm cryopreservation: why isn't it better? Theriogenology. 57, 327-344.
Nally W.M.M., Handarani R., Yusuf T.L., Purwantra P. and Semiadi G. (2010). The effect of glycerol concentration in tris glucose egg yolk extender on the quality of timor deer frozen semen. J. Indonesian Trop. Anim. Agric. 36(2), 91-96.
Neild D.M., Brouwers J.P., Colenbrander B., Aguero A. and Gadella B.M. (2005). Lipid peroxide formation in relation to membrane stability of fresh and frozen-thawed stallion spermatozoa. Mol. Reprod. Dev. 72, 230-238.
Ozkavukcu S., Erdemli E., Isik A., Oztuna D. and Karahuseyinoglu S. (2008). Effects of cryopreservation on sperm parameters and ultrastructural morphology of human spermatozoa. J. Assist. Reprod. Genet. 25, 403-411.
Pramanik P.S. and Raina V.S. (2001). HOS response and post-thaw incubation motility of buffalo spermatozoas in various extenders. Indian Vet. Med. J. 25, 129-131.
Priyadharsini R., Jindal S.K., Sharma D., Ramachandran N., Karche S.D. and Goel A.K. (2011). Effect of different egg yolk level on the cryopreservation capability of Jakhrana goat semen. J. Anim. Sci. Adv. 1(1), 28-37.
Purdy P.H. (2006). A review on goat sperm cryopreservation. Small Rumin. Res. 63, 215-225.
Rahman A.N.M.A., Abdullah R.B. and Khadijah W.E.W. (2008). A review of reproductive biotechnologies and their application in goat. Biotechnology. 7(2), 371-384.
Ranjan R., Ramchandran N., Jindal S.K., Goel A.K. and KharcheS.D. (2009). Effect of egg yolk levels on keeping quality of Marwari buck semen at refrigeration temperature. Indian J. Anim. Sci. 79(7), 662-664.
Rao C.K. (1957). Different staining of live and dead sperm. Indian Vet. J. 29, 307-309.
Ritar A.J., Ball P.D. and O'May P.J. (1990). Examination of methods for the deep freezing of goat semen. Reprod. Fertil. Dev. 2, 27-34.
Salamon S. and Maxwell W.M.C. (2000). Storage of ram semen. Anim. Reprod. Sci. 62, 71-111.
Sarma P.V. (1995). A simplified staining technique for evaluation of acrosomal status of sperm cells. J. Anim. Sci. 16, 127-127.
Shamsuddin M., Amiri Y. and Bhuiyan M.M.U. (2000). Characteristic of buck semen with regards to ejaculate numbers, collection intervals, dilution and preservation periods. Reprod. Domest. Anim. 35, 53-57.
SPSS Inc. (2011). Statistical Package for Social Sciences Study. SPSS for Windows, Version 20. Chicago SPSS Inc.
Stanger J.D., Long V., Yovich J.L. and Almahbobi G. (2010). Hypo osmotic swelling test identifies individual spermatozoa with minimal DNA fragmentation. Reprod. Biomed. Online. 21, 474-484.
Sundararaman M.N. and Edwin M.J. (2005). Evaluation of sperm motion characteristics of pre-freeze and post-thaw goat spermatozoa on computer assisted semen analysis (CASA). Indian J. Small Rumin. 11, 92-95.
Watson P.F. (1975). The interaction of egg yolk and ram spermatozoa studied with fluorescent probe. J. Reprod. Fertil. 42, 105-111.
Watson P.F. (1976). The protection of ram and bull spermatozoa by the low density lipoprotein fraction of egg yolk during storage at 5 ˚C and deep-freezing. J. Thermal. Biol. 1, 137-141.
Watson P.F. (1995). Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod. Fertil. Dev. 7, 871-891.
Watson P.F. (2000). The causes of reduced fertility with cryopreserved semen. Anim. Reprod. Sci. 60, 481-492.
Yimer N., Noraisyah A.H., Rosnina Y., Wahid H., Sarsaifi K. and Hafizal A.M. (2014). Comparison of cryopreservative effect of different levels of omega-3 egg-yolk in citrate extender on the quality of goat spermatozoa. Pakistan Vet. J. 34(3), 347-350.
