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Influence of Osmolality of Complete Semen Extender on Motion Characteristics of Frozen-thawed Ram Spermatozoa

  • Joshi, Anil (Semenology laboratory, Division of Physiology and Biochemistry, Central Sheep & Wool Research Institute Avikanagar) ;
  • Mathur, A.K. (Project Directorate on Cattle) ;
  • Naqvi, S.M.K. (Semenology laboratory, Division of Physiology and Biochemistry, Central Sheep & Wool Research Institute Avikanagar) ;
  • Mittal, J.P. (Semenology laboratory, Division of Physiology and Biochemistry, Central Sheep & Wool Research Institute Avikanagar)
  • Received : 2005.07.21
  • Accepted : 2005.10.19
  • Published : 2005.12.01

Abstract

The present study was conducted to observe the effect of osmolality of glycerolated TEST-yolk glycerol extenders on post-thawing sperm kinematics of ram spermatozoa of the native Malpura breed maintained in a semi-arid tropical environment. Good quality semen obtained from adult rams was pooled, split and diluted to 1,000 million spermatozoa per ml in complete TEST-yolk-glycerol extenders of 900, 1,200, 1,500 and 1,800 mOsm/kg osmolality. Diluted semen samples were loaded in 0.25 ml straws and cooled down to $-125^{\circ}C$ freezing temperature at the rate of $-25^{\circ}C$ per minute under controlled conditions before plunging into liquid nitrogen for storage. The thawing of straws was performed at $50^{\circ}C$ in a water bath for 10 seconds and sperm kinematics of the frozen-thawed spermatozoa were assessed by a computer-assisted sperm analysis technique. Osmolality of diluent had no significant effect on post-thawing % motility, % rapid, % medium and % slow moving frozen-thawed spermatozoa but significantly (p< 0.05) affected the % linearity and % straightness. The post-thawing % motility and % rapid motile spermatozoa were highest in samples extended in diluent of 1,500 mOsm/kg osmolality and lowest in 900 mOsm/kg. The curvilinear velocity of spermatozoa was significantly (p<0.05) higher for samples extended in 1,800 mOsm/kg, compared to those in 900 and 1,200 mOsm/kg, but the effect was not significantly different to those extended in diluent of 1,500 mOsm/kg osmolality. The study indicated that ram spermatozoa could tolerate a wide osmolality range for dilution in the complete TEST-yolk-glycerol extender for their cryosurvival. The highest recovery of motile spermatozoa following thawing was achieved in samples extended in the TEST-yolk-glycerol diluent of 1,500 mOsm/kg osmolality.

Keywords

Sheep;Extender;Osmolality;Spermatozoa;Cryopreservation;Computer-aided Semen Analysis

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Cited by

  1. Effect of Controlled and Uncontrolled Rate of Cooling, Prior to Controlled Rate of Freezing, on Motion Characteristics and Acrosomal Integrity of Cryopreserved Ram Spermatozoa vol.6, pp.4, 2008, https://doi.org/10.1089/bio.2008.0013