Acta Vet. Brno 2015, 84: 383-391

The effect of the freezing curve type on bull spermatozoa motility after thawing

Martina Doležalová, Luděk Stádník, Zuzana Biniová, Jaromír Ducháček, Jan Beran

Czech University of Life Sciences Prague, Faculty of Agrobiology, Food and Natural Resources, Department of Animal Husbandry, Czech Republic

Received February 24, 2015
Accepted June 24, 2015

The objective of this work was to determine the effect of selected freezing curves on spermatozoa survivability after thawing, defined by its motility. The ejaculates of nine selected sires of the same age, breed, and frequency of collecting, bred under the same breeding conditions including handling, stabling, feeding system and feeding ratio composition, were repeatedly collected and evaluated. Sperm samples of each sire were diluted using only one extender and divided into four parts. Selected four freezing curves – the standard, commercially recommended three-phase curve; a two-phase curve; a slow three-phase curve; and a fast three-phase curve, differing in the course of temperature vs time, were applied. The percentage rate of progressive motile spermatozoa above head was determined immediately after thawing, and after 30, 60, 90, and 120 min of the thermodynamic test (TDT). Moreover, average spermatozoa motility (AMOT) and spermatozoa motility decrease (MODE) throughout the entire TDT were evaluated. Insemination doses frozen using the simpler two-phase curve demonstrated the highest motility values (+2.97% to +10.37%; P < 0.05–0.01) immediately after thawing and during the entire TDT. Concurrently, the highest AMOT (+4.37% to +8.82%; P < 0.01) was determined. The highest spermatozoa motility values were detected after thawing doses frozen by the two-phase freezing curve in eight out of nine sires. Simultaneously, a significant effect of sire individuality was clearly confirmed. Inter-sire differences of spermatozoa motility during TDT as well as AMOT and MODE were significant (P < 0.01). The findings describing both factors of interaction indicate the necessity of individual cryopreservation of the ejaculate to increase its fertilization capability after thawing.


The work was funded by the “S” grant of the Ministry of Education, Youth and Sports of the Czech Republic and project NAZV QJ1210109.


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