Research article - Peer-reviewed, 2003
Evaluation of cryopreserved stallion semen from Tori and Estonian breeds using CASA and flow cytometry
Kavak A, Johannisson A, Lundeheim N, Rodriguez-Martinez H, Aidnik M, Einarsson SAbstract
Methods to evaluate the quality of frozen-thawed stallion semen are still needed, particularly those considering the sperm function. The present study evaluated sperm motility, membrane and acrosome integrity and the capacitation status of frozen-thawed spermatozoa from seven Tori and six Estonian breed stallions by way of computer assisted sperm analysis (CASA), a triple fluorophore stain combination and Merocyanine 540, respectively, the latter ones using flow cytometry. Two ejaculates from each stallion were cryopreserved using the Hannover method in 0.5 ml plastic straws. Two straws per ejaculate per stallion were thawed at 37degreesC for 30 s. Motility was analysed with CASA immediately after thawing, while for flow cytometry spermatozoa were cleansed by 70:40% Percoll discontinuous density gradient separation before analysed for sperm viability, acrosome integrity (stained with SNARF, PI and FITC-PSA) and capacitation status (stained with Merocyanine 540/Yo-Pro-1). Results (as least square means) were as follows: the motility of frozen-thawed semen was 43.4% for Tori stallions and 42.3% for Estonian stallions (P > 0.05). After Percoll separation 79.3% of the spermatozoa from Tori stallions had intact acrosomes and 1.7% of them showed early signs of capacitation. The same parameters for Estonian stallions were 84.5 and 2.3%, respectively. There were no statistically significant differences between breeds or ejaculates within breed for any evaluated parameter. We conclude that triple staining and flow cytometry are valuable techniques to evaluate frozen-thawed stallion spermatozoa, and that no differences in quality of frozen semen were registered between Tori and Estonian breed stallions, allowing implementation of this technology in the Estonian horse population. (C) 2002 Elsevier Science B.V. All rights reservedPublished in
Animal Reproduction Science2003, volume: 76, number: 3-4, pages: 205-216
Publisher: ELSEVIER SCIENCE BV
Authors' information
Swedish University of Agricultural Sciences, Department of Animal Breeding and Genetics
Aidnik, M
Kavak, A.
Swedish University of Agricultural Sciences, Department of Anatomy, Physiology and Biochemistry (AFB)
Rodriguez-Martinez, H
Einarsson, Stig
UKÄ Subject classification
Animal and Dairy Science
Veterinary Science
Publication Identifiers
DOI: https://doi.org/10.1016/S0378-43200(02)00247-6
URI (permanent link to this page)
https://res.slu.se/id/publ/538