Effects of Rapid Cooling Prior to Freezing on the Quality of Canine Cryopreserved Spermatozoa |
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| Authors: | Carmen RODENAS Inmaculada PARRILLA Jordi ROCA Emilio Arsenio MARTINEZ Xiomara LUCAS |
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| Institution: | 1)Department of Animal Medicine and Surgery, University of Murcia, Espinardo 30100, Spain |
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| Abstract: | The aim of this study was to evaluate the effects of rapid cooling prior to freezing on frozen-thawed canine sperm quality.
In experiment 1, centrifuged ejaculates from 6 dogs were pooled, split into 4 aliquots and cryopreserved by the Uppsala
procedure using different cooling rates (control, cooling speed 18 C/90 min and average cooling rate 0.2 C/min; rapid,
cooling speed 18 C/8 min and average cooling rate 2.25 C/min) in combination with 2 glycerol addition protocols (fractionated
or unfractionated). In experiment 2, centrifuged ejaculates from 4 dogs were processed individually using the same cooling
rates described in experiment 1 in combination with an unfractionated glycerol addition protocol. Each of the experiments was
replicated 5 times. Sperm quality was evaluated after 30 and 150 min of post-thawing incubation at 38 C. Total motility (TM),
progressive motility (PM) and quality of movement parameters were assessed using a computerized system, and sperm viability
(spermatozoa with intact plasma and acrosome membranes) was assessed using flow cytometry (H-42/PI/FITC-PNA). Values for TM,
PM, viable spermatozoa and the quality of movement parameters after thawing were not significantly affected by the cooling
rate. The interaction between the cooling rate and the added glycerol protocol was not significant. There were significant
differences among the males (P<0.01) in the sperm quality parameters evaluated after thawing. The interaction between the
males and the cooling rate was not significant. In conclusion, canine spermatozoa can be cryopreserved using the Uppsala
method at an average cooling rate of 2.25 C/min prior to freezing together with addition of fractionated or unfractionated
glycerol. |
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| Keywords: | Canine Cooling-rate Cryopreservation Spermatozoa |
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