title: Equine sperm post-thaw evaluation after the addition of different cryoprotectants added to INRA 96® extender.
abstract: The rise of assisted reproduction techniques in equine medicine has fostered investigations that seek to optimize methods to increase fertility rates. Since cryopreservation continues to give low values of viability in stallions, the handling and preservation of the sperm is of vital importance. This reduction of fertility makes it essential for farmers to find new options that ensure reliability in the use of these techniques. The main objective of this study was to assess the effect of INRA 96® (manufactured commercial extender for cooling of Equine semen) as an extender for cryopreservation in combination with different cryoprotectants: Acetal (5%), Dimethylformamide (5%) and Glycerol (5%), alone and combined (2.5% each) on ejaculated and epididymal spermatozoa. Ejaculates collected from mature stallion and epididymal sperm samples were cryopreserved in INRA® varying content of cryoprotectant and cryopreserved. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. We conclude that INRA 96® is suited as extender for freezing when it is used in combination with Dimethylformamide (5%) or Dimethylformamide (2.5%)+Glycerol (2.5%) for samples of ejaculate. The combination of Dimethylformamide (2.5%)+Glycerol (2.5%) showed the best results on epididymal spermatozoa. In conclusion, the combination of Dimethylformamide and Glycerol as cryoprotectants in INRA® medium enhanced equine epididymal and ejaculated spermatozoa quality after cryopreservation.
authors.affiliation: Centro Militar de Cría Caballar de Zaragoza, Spain. Electronic address: calvsan@gmail.com.
authors.forename: C
authors.initials: C
authors.lastname: Álvarez
authors.affiliation: Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Gil
authors.affiliation: Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
authors.forename: N
authors.initials: N
authors.lastname: González
authors.affiliation: Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Olaciregui
authors.affiliation: Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
authors.forename: V
authors.initials: V
authors.lastname: Luño
mesh_terms: D000177:Acrosome / Q000502:physiology; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004126:Dimethylformamide / Q000494:pharmacology*; D004822:Epididymis / Q000166:cytology; D005990:Glycerol / Q000031:analogs & derivatives / Q000494:pharmacology*; D006736:Horses; D008297:Male; D009997:Osmotic Pressure / Q000187:drug effects; D015203:Reproducibility of Results; D012661:Semen; D055101:Semen Analysis; D012662:Semen Preservation / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects
chemical_list: D003451:Cryoprotective Agents; D004126:Dimethylformamide; D005990:Glycerol
keywords: Acetal; Cryopreservation; Dimethylformamide; Equine sperm; Glycerol