title: Use of single-layer centrifugation with Androcoll-C to enhance sperm quality in frozen-thawed dog semen.
abstract: The aim of this study was to investigate whether single-layer centrifugation (SLC) with Androcoll-C could select good quality spermatozoa, including those with specific motility patterns, from doses of frozen dog semen. Semen from five dogs was collected and cryopreserved following a standard protocol. After thawing, the semen samples were divided in two aliquots, one of which was used as a control and the other one processed by SLC. Assessment of sperm motility (assessed by computer-assisted semen analysis), morphology (Diff-Quick staining), viability (dual staining with propidium iodine/acridine orange), and acrosome integrity (dual staining with propidium iodine/isothiocyanate-labeled peanut [Arachis hypogaea] agglutinin) were performed on aliquots of fresh semen, frozen-thawed control samples, and frozen-thawed SLC-treated preparations. A multivariate clustering procedure separated 57,577 motile spermatozoa into three subpopulations (sP): sP1 consisted of poorly active and nonprogressive spermatozoa (48.8%), sP2 consisted of moderately slow but progressive spermatozoa (13.3%), and sP3 consisted of highly active and/or progressive spermatozoa (37.8%). SLC with Androcoll-C yielded sperm suspensions with improved motility, viability, and acrosome integrity (P < 0.01). The frozen-thawed SLC-treated samples were enriched in sP3, representing 38.5% of the sperm population. Likewise, sP2 was more frequently observed after SLC, but not significantly so. From these results, we concluded that for dog semen samples selected by SLC with Androcoll-C after thawing, the sperm quality parameters, including motility patterns, are better than in frozen-thawed control samples.
authors.affiliation: Animal Reproduction Group, Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Cordoba, Córdoba, Spain. Electronic address: jdorado@uco.es.
authors.forename: J
authors.initials: J
authors.lastname: Dorado
authors.forename: M J
authors.initials: MJ
authors.lastname: Gálvez
authors.forename: J M
authors.initials: JM
authors.lastname: Morrell
authors.forename: L
authors.initials: L
authors.lastname: Alcaráz
authors.forename: M
authors.initials: M
authors.lastname: Hidalgo
mesh_terms: D000177:Acrosome / Q000648:ultrastructure; D000818:Animals; D002498:Centrifugation / Q000379:methods / Q000662:veterinary*; D016000:Cluster Analysis; D015925:Cryopreservation; D004285:Dogs; D008297:Male; D015999:Multivariate Analysis; D012661:Semen / Q000166:cytology*; D013081:Sperm Motility
keywords: Cryopreservation; Dog spermatozoa; Motility patterns; Sperm quality; Sperm separation