Analysis of Awassi Sperm Motility in Two Media at Different Levels of Temperature, pH and Osmolality
Subject Areas : Camelم. آلومار 1 * , م. زرکاوی 2 , م.آ. آلزوآبی 3
1 - Department of Agriculture, Atomic Energy Commission, Damascus, Syria
2 - Department of Agriculture, Atomic Energy Commission, Damascus, Syria
3 - Department of Agriculture, Atomic Energy Commission, Damascus, Syria
Keywords: temperature, ram, computer-aided sperm analyzer, osmolality,
Abstract :
The effects of five levels of osmolality and pH as well as three temperatures on Awassi ram sperm motility parameters assessed by computer-aided sperm analyzer(CASA)in tyrode albumin lactate pyruvate (TALP) and egg-yolk-tris (YET) media were studied. Semen obtained from five adult Awassi rams were pooled and diluted in the above two media at 100, 200, 300, 400 and 500 mOsm/kg at temperatures of 4˚, 20˚ and 37˚ degree celsius and also at pH levels of 5, 6, 7, 8 and 9 at the same temperatures. The osmolality, pH and temperature had a significant effect (P<0.05) on the percent motility (MOT %) and the percent of sperm showing progressive motility (PMOT %). With the exception of lateral head displacement, all the other CASA motility parameters were also significantly affected (P<0.05). Semen incubated in YET was able to tolerate osmolarities within 200 to 400 mOsm/kg range for MOT %. Alkaline condition at pH 9 had higher negative effect than acidic condition at pH 5. Compared to the 20 ˚C and 37 ˚C, the 4 ˚C had negatively affected MOT % and PMOT % and this was more obvious in TALP medium. In conclusion, our results suggest: 1) the significant effects of osmolality, pH and temperature on Awassi sperm incubated in both TALP and YET media and 2) the need for careful selection of temperature by which the semen may be manipulated during artificial insemination and in vitro fertilization experiments in these two media.
Ak K., Cirit U., Nur Z., Bacinoglu S., Pabuccuoglu S., Ozdas O.B. and Birler S. (2010).Effects of extender osmolarity, cooling rate, dilution rate and glycerol addition time on post-thaw ram semen characteristics and fertilization. J. Fac. Vet. Med. Istanbul Univ. 36, 33-46.
Alomar M., Mahieu J., Verhaeghe B., Defoin L. and Donnay I. (2006). Assessment of sperm quality parameters of six bulls showing different abilities to promote embryo development. Reprod. Fertil. Dev. 18, 395-402.
Amann R.P. and Waberski D. (2014). Computer-assisted sperm analysis (CASA): Capabilities and potential developments. Theriogenology. 81, 5-17.
Bag S., Joshi A., Rawat P.S. and Mittal J.P. (2002a).Effect of initial freezing temperature on semen characteristics of frozen-thawed ram spermatozoa in a semi-arid tropical environment. Small Rumin. Res. 43, 23-29.
Bag S., Joshi A., Naqvi S.M.K., Rawat P.S. and Mittal J.P. (2002b). Effect of freezing temperature, at which straws were plunged into liquid nitrogen, on the post-thaw motility and acrosomal status of ram spermatozoa. Anim. Reprod. Sci. 72, 175-183.
Carr D.W. and Acott T.S. (1989). Intracellular pH regulates bovine sperm motility and protein phosphorylation. Biol. Reprod. 41, 907-920.
Comhaire F.H., Huysse S., Hinting A., Vermeulen L. and Schoonjans F. (1992). Objective semen analysis: Has the target been reached? Hum. Reprod. 7, 237-241.
Contri A., Gloria A., Robbe D., Valorz, C., Wegher L. and Carluccio A. (2013). Kinematic study on the effect of pH on bull sperm function. Anim. Reprod. Sci. 136, 252-259.
Curry M.R., Millar J.D. and Watson P.F. (1994).Calculated optimal cooling rates for ram and human sperm cryopreservation fail to conform with empirical observations. Biol. Reprod. 51, 1014-1021.
Drobnis E.Z., Crowe L.M., Berger T., Anchordoguy T.J., Overstreet J.W. and Crowe J.H. (1993).Cold shock damage is due to lipid phase-transitions in cell-membranes a demonstration using sperm as a model. J. Exp. Zool. 265, 432-437.
Eppleston J., Salamon S., Moore N.W. and Evans G. (1994).The depth of cervical insemination and site of intrauterine insemination and their relationship to the fertility of frozen-thawed ram semen. Anim. Reprod. Sci. 36, 211-225.
Farrell P.B., Foote R.H., Mcardle M.M., Trouern-Trend V.L. and Tardif A.L. (1996).Media and dilution procedures tested to minimize handling effects on human, rabbit and bull sperm for computer-assisted sperm analysis (CASA). J. Androl. 17, 293-300.
Farrell P.B., Presicce G.A., Brockett C.C. and Foote R.H. (1998). Quantification of bull sperm characteristics measured by computer-assisted sperm analysis (CASA) and the relationship to fertility. Theriogenology. 49, 871-879.
Galal S., Gursoy O. and Shaat I. (2008). Awassi sheep as a genetic resource and efforts for their genetic improvement: A review. Small Rumin. Res. 79, 99-108.
Herrara C., Brogliatti G., Cavia R., Conde P., Revora M. and Pasqualini R.S. (2005). CASA sperm parameters and their relation with in vitro fertilization. Pp. 411-415 in Proc. 15th Int. Congr. Anim. Reprod., Porto Seguro, Brazil.
Holt W.V. (2000). Basic aspects of frozen storage of semen. Anim. Reprod. Sci. 62, 3-22.
Holt W.V. and Palomo M.J. (1996).Optimization of a continuous real-time computerized semen analysis for ram sperm motility assessment, and evaluation of four methods of semen preparation. Reprod. Fertil. Dev. 8, 219-230.
Joshi A., Mathur A.K., Naqvi S.M.K. and Mittal J.P. (2006).Influence of osmolality of complete semen extender on motion characteristics of frozen-thawed ram spermatozoa. Asian-Australasian J. Anim. Sci. 19, 716-1721.
Langlais J. and Roberts K.D. (1985).A molecular membrane model of sperm capacitation and the acrosome reaction of mammalian spermatozoa. Gamete Res. 12, 183-224.
Makler A. and Jakobi P. (1981). Factors affecting sperm motility. V. Washing and resuspension of human spermatozoa in various artificial media. Fertil. Steril. 35, 442-446.
Makler A., David R., Blumenfeld Z. and Better O.S. (1981). Factors affecting sperm motility. VII sperm viability as affected by changes of pH and osmolarity of semen and urine specimens. Fertil. Steril. 36, 507-511.
Mortimer S.T. (1997). A critical review of the physiological importance and analysis of sperm movement in mammals. Hum. Reprod. Update. 3, 403-439.
Mortimer S.T. (2000). CASA-Practical aspects. J. Androl. 21, 515-524.
Mortimer D. and Mortimer S.T. (2013). Computer-aided sperm analysis (CASA) of sperm motility and hyperactivation. Methods Mol. Biol. 927, 77-87.
Nur Z., Zik B., Ustuner B., Sagirkaya H. and Ozguden C.G. (2010). Effects of different cryoprotective agents on ram sperm morphology and DNA integrity. Theriogenology. 73, 1267-1275.
Rijsselaere T., Mase D., Hoflack G., de Kruif A. and Van Soom A. (2007). Effect of body weight, age and breeding history on canine sperm quality parameters measured by the Hamilton-Thorne Analyser. Reprod. Domest. Anim. 42, 143-148.
Robayo I., Montenegro V., Valdes C. and Cox J.F. (2008). CASA assessment of kinematic parameters of ram spermatozoa and their relationship to migration efficiency in ruminant cervical mucus. Reprod. Domest. Anim. 43, 393-399.
Salamon S. and Maxwell W.M.C. (1995a). Frozen storage of ram semen. I. Processing, freezing, thawing and fertility after cervical insemination (Review). Anim. Reprod. Sci. 37, 185-249.
Salamon S. and Maxwell W.M.C. (1995b). Frozen storage of ram semen. II. Causes of low fertility after cervical insemination and methods of improvement. Anim. Reprod. Sci. 38, 1-36.
Sleiman F.T. and AbiSaab S. (1995). Influence of environment on respiration, heart rate and body temperature of filial crosses compared to local Awassi sheep. Small Rumin. Res. 16, 49-53.
Vantman D., Bands S.M., Koukoulis G., Dennison L. and Sherins R.J. (1989).Assessment of sperm motion characteristics from fertile and infertile men using a fully automated computer-assisted semen analyzer. Fertil. Steril. 51, 156-161.
Varisli O., Uguz C., Agca C. and Agca Y. (2009). Motility and acrosomal integrity comparisons between electro-ejaculated and epididymal ram sperm after exposure to a range of anisosmotic solutions, cryoprotective agents and low temperatures. Anim. Reprod. Sci. 110, 256-268.
Visconti P.E., Westbrook V.A., Chertihin O., Demarco I., Sleight S. and Diekman A.B. (2002).Novel signaling pathways involved in sperm acquisition of fertilizing capacity. J. Reprod. Immunol. 53, 133-150.
Watson P.F. (1975). The interaction of egg yolk and ram spermatozoa studied with a fluorescent probe. J. Reprod. Fertil. 42, 105-111.