Document Type : Original Article

Authors

1 M.Sc. Student of Animal Physiology, Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran

2 Professors, Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran

Abstract

Semen collection evaluation and addition of preservatives to increase storage period of sperm are essential for successful artificial insemination. This study was conducted on 4rams (2 Ghezel Merinos and 2 Merinos Moghani) to evaluate the effect of two diluents in Khalatposhan research station. Average age of rams was 3 years. Rams were trained to serve the artificial insemination and semen samples were collected weekly. It was started from October 2011 to June 2012. After collection semen samples were mixed with diluents. They were stored in liquid nitrogen. After diluting and storage semen was assessed after 0, 1, 2 and 3 day for pH, viability and progressive motility. Frozen straws were thawed individually at 37 °C for 20 s in a water bath for evaluation. Effect of diluent and storage day on pH, viability and progressive motility of sperm were significant (P< 0.01). With the increasing storage day pH, viability and progressive motility of sperm decreased. The success of these diluent has been attributed the tris which may act as a buffer against changes in pH and tonicity. Fructose and citric acid are energy source. Egg- yolks protect the cell membrane during cooling. Glycerol protects the spermatozoa against membrane damage during freezing. This study showed that diluents containing of 7% glycerol and 20% egg-yolk had better sperm protection ability than extender containing 5% glycerol and 5% egg-yolk  according to sperm motility, pH and sperm viability

Keywords

Amann R, Pickett B (1987). Principles of cryopreservation and a review of stallion spermatozoa. Equine. Vet. Sci. 7, 145–173.
Avdi M, Leboeuf B, TerquiM (2004). Advanced breeding and buck effect in indigenous Greek goats. Livestock. Prod. Sci. 87, 251–257.
Blackburn HD (2004). Development of national animal genetic resource programs. Reprod. Fertil. Dev. 16, 27–32.
Colas G (1975). Effect of initial freezing temperature, addition of glycerol and dilution on the survival and fertilizing ability of deep-freezing ram semen. J. Reprod. Fertil. 42, 277–285.
Evanc G, Salmons WMC (1987). Artificial insemination of sheep and goats. Star. Printer. pty ltd. 108- 145.
Fisher PS, Fairfull RW (1984). The effect of glycerol concentration and cooling velocity on cryosurvival of ram spermatozoa frozen in straws. Cryobiology. 21, 542–551.
Graham EF, Crabo BG, Pace MM ( 1978). Current status of semenpreservation in the ram, boar andstallion. J. Anim. Sci. 2, 80–119.
Hammerstedt RH, Graham JK (1992). Cryopreservation of poultry sperm. the enigma of glycerol. Cryobiology. 29, 26–38
Isachenko E (2003). Vitrification of mammalian spermatozoa in the absence of cryoprotectans, from past practical difficulties to present. Reprod. Biol. 6, 191–200.
Jeyendran R, Acosta V, Land S, Coulam C (2008). Cryopreservation of human sperm in a lecithinsupplemented freezing medium. Fertil. Steril. 90, 1263–5.
Matsuoka T, Imai H, Kohno H, Fukui Y (2006). Effects of bovine serum albumin and trehalose in semen diluents for improvement of frozenthawed ram spermatozoa. J. Reprod. Dev. 52, 675–683.
Moussa M, Matinet V, Trimeche A, Tainturier D, Anton M (2002) . Low density lipoproteins extracted from hen egg yolk by an easy method. cryoprotective effect on frozen-thawed bull semen. Theriogenology. 57, 1695–706.
Paulenz H, Soderquist L, Perez-Pe R, Berg KA (2002). Effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriogenology. 57, 823–36.
Pontbriand D, Howard JG, Schiewe MC, Stuart LD, WildtDE (1989). Effect of cryoprotective diluent and method of freeze-thawing on survival and acrosomal integrity of ram spermatozoa. Cryobiology. 26, 341–54.
Preservage S, Hassan M R, Ershaduzzaman M, Khandoker M A M Y (2009) .Preservation of liquid semen and artificial insemination in native sheep. J. Bangladesh Agri. Univer. 7, 305-308.
Purdy PH (2006). A review on goat sperm cryopreservation.Small. Rumin. Res. 63, 215–225.
Rodriguez-Martinez H, Barth AD (2007). In vitro evaluation of sperm quality related to in vivo function and fertility. Soc. Reprod. Fertil. Suppl. 64, 39–54.
Salamon S, Lightfoot R J (1969). Freezing of ram spermatozoa by the pellet method. I. The effect of diluentcomposition on survival of spermatozoa. Aust. J. Biol. Sci. 22, 1527–1546.
Salamon S, Maxwell W.M.C (1995). Frozen storage of ram semen.Processing, freezing, thawing and fertility after cervical insemination. Anim. Reprod. Sci. 37, 185-249.
Salamon S, Maxwell WM ( 2000). Storage of ram semen. Anim. Reprod. Sci. 62, 77-111.
Shamsuddin M, Amiri Y, Bhuiyan MMU (2000). Characteristics of buck Semen with regard to ejaculate numbers, collection intervals, diluents and preservation periods. Reprod. Dom. Anim. 35, 53-57.
Watson PF, Martin ICA (1975). Effects of egg yolk, glycerol and the freezing rate on the viability and acrosomal structures of frozen ram spermatozoa. Aust. J. Biol. Sci. 28, 153-159.
Waston PF, Martin CA (1975). The influence of some fractions of egg yolk on the survival of ram spermatozoa at 5 8C. Aust. J. Biol . Sci 28, 145-52.
Waston PF (1981). The role of lipid and protein in the protection of ram spermatozoa at 5 °C by egg yolk lipoprotein. J. Reprod. Fertil. 2, 337-40.
Watson PF (1995). Recent development and concepts in the conservation of spermatozoa and the assessment of their post-thawing function. Reprod. Fertil. Dev. 7, 871-891.