Molecular Biology Reports

, Volume 38, Issue 3, pp 1499–1506

Freeze–thaw induced genotoxicity in buffalo (Bubalus bubalis) spermatozoa in relation to total antioxidant status


DOI: 10.1007/s11033-010-0257-1

Cite this article as:
Kumar, R., Jagan Mohanarao, G., Arvind et al. Mol Biol Rep (2011) 38: 1499. doi:10.1007/s11033-010-0257-1


Use of cryopreserved semen has become an important tool in assisted reproduction but freezing and thawing cause sub-lethal damage to spermatozoa. This is detrimental to sperm because of the membrane damage including permeability and integrity. An excess generation of reactive oxygen species (ROS) creates oxidative stress due to reduced antioxidant status of the cryopreserved spermatozoa. In the present study fresh buffalo semen was collected and divided into two aliquots. One aliquot was used for fresh semen analysis and the other was cryopreserved in Tris-egg yolk-citrate extender. The semen samples were used to study different sperm quality parameters like motility, viability, membrane integrity and total antioxidant status. The DNA integrity in fresh and cryopreserved spermatozoa was also studied using comet assay. The sperm quality parameters like post-thaw sperm motility, viability, membrane integrity and total antioxidant status of cryopreserved spermatozoa were significantly lowered (P < 0.05) compared to fresh spermatozoa. The DNA fragmentation in cryopreserved spermatozoa was significantly higher (P < 0.01) as compared to fresh spermatozoa. The results show that the irreversible DNA damage occurs in spermatozoa during cryopreservation.


BuffaloComet assayCryopreservationDNA damageSpermatozoaTotal antioxidant status



Reactive oxygen species


Deoxyribonucleic acid

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Animal Biochemistry DivisionNational Dairy Research InstituteKarnalIndia