Abstract
Aim of study: There is little information about vitrification of sperm in large volumes (up to 0.5 mL). This study aimed to develop the vitrification technique in 0.5 mL straws in donkey sperm, evaluating the effect of three warming temperatures.Area of study: Cordoba, Spain.Material and methods: Ejaculates from five donkeys were divided in four groups: one control subjected to conventional slow freezing (C) and three vitrified in 0.5 mL straws and warmed using different protocols (W1: 37ºC/30s, W2: 43ºC/20s and W3: 70ºC/8s+37ºC/52s). Sperm motility, kinematic parameters, plasma membrane and acrosome integrity were evaluated. Conventional freezing resulted in significantly higher values for total (42.7±19.6%), and progressive motility (30.3±16.7%), plasma membrane (49.1±10.4%) and acrosome integrity (39.6±14.5%) respect to vitrification method.Main results: Values after warming ranged between 0.2-2.8% for total motility; 0.2-2.1% for progressive motility; 5.5-20.0% for plasma membrane integrity and 14.5-29.8% for acrosome integrity in all warming protocols after sperm vitrification. However, no differences were found between W3 and C for kinematic parameters; and W3 resulted in significantly higher values for membrane integrity (20.0±11.0%) in comparison to W1 (5.5±3.6%) and W2 (9.3±8.4%).Research highlights: High warming rates seem to be better for donkey sperm vitrification in large volumes; but this methodology is still not an alternative to conventional sperm freezing.
Highlights
Since sperm vitrification in straws was first described in human by Isachenko et al (2005), it has been investigated in different animals including ram (Jiménez-Rabadán et al, 2015), goat (Daramola et al., 2016), fish (Zilli et al, 2018) and horse (Consuegra et al, 2018; Restrepo et al, 2019)
Research highlights: High warming rates seem to be better for donkey sperm vitrification in large volumes; but this methodology is still not an alternative to conventional sperm freezing
Since sperm vitrification in straws was first described in human by Isachenko et al (2005), it has been investigated in different animals including ram (Jiménez-Rabadán et al, 2015), goat (Daramola et al, 2016), fish (Zilli et al, 2018) and horse (Consuegra et al, 2018; Restrepo et al, 2019)
Summary
Since sperm vitrification in straws was first described in human by Isachenko et al (2005), it has been investigated in different animals including ram (Jiménez-Rabadán et al, 2015), goat (Daramola et al., 2016), fish (Zilli et al, 2018) and horse (Consuegra et al, 2018; Restrepo et al, 2019). This technique has been developed in donkey sperm in absence of permeable cryoprotectants, showing promising. To our knowledge, there is little information about its use in animals, and if so, no motile sperm was recovered after warming (Restrepo et al, 2019)
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