title: Centrifugation on PureSperm(®) density-gradient improved quality of spermatozoa from frozen-thawed dog semen.
abstract: The main objective of this study was to investigate if centrifugation through PureSperm(®) density-gradient can improve the post-thaw semen quality of dog semen. Semen from 5 dogs was collected and cryopreserved following a standard protocol. After thawing, semen samples were selected by centrifugation on PureSperm(®). Assessments of sperm motility (assessed by computerized-assisted semen analysis), morphology (Diff-Quick staining) and viability (triple fluorescent stain of Propidium iodine/isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin/Rhodamine 123), were performed on aliquots of fresh semen, unselected samples and selected preparations. Cryopreservation had a significant (P < 0.001) effect on all studied semen parameters. PureSperm(®) centrifugation yielded sperm suspensions with improved motility and viability (P < 0.001). The washing step significantly reduced (P < 0.001) all of the kinematics parameters evaluated as well as reduced the proportion of viable spermatozoa with intact acrosomes (P < 0.05). We concluded that PureSperm(®) centrifugation is a successful method for improving the quality of frozen-thawed dog spermatozoa. However, washing after density-gradient centrifugation dramatically reduces the post-thaw semen quality, indicating that the inclusion of such a washing step is unnecessary.
authors.affiliation: Department of Medicine and Animal Surgery, University of Cordoba, Córdoba, Spain. jdorado@uco.es
authors.forename: J
authors.initials: J
authors.lastname: Dorado
authors.forename: L
authors.initials: L
authors.lastname: Alcaráz
authors.forename: N
authors.initials: N
authors.lastname: Duarte
authors.forename: J M
authors.initials: JM
authors.lastname: Portero
authors.forename: D
authors.initials: D
authors.lastname: Acha
authors.forename: S
authors.initials: S
authors.lastname: Demyda
authors.forename: A
authors.initials: A
authors.lastname: Muñoz-Serrano
authors.forename: M
authors.initials: M
authors.lastname: Hidalgo
mesh_terms: D000177:Acrosome / Q000502:physiology; D000818:Animals; D002470:Cell Survival; D002499:Centrifugation, Density Gradient / Q000379:methods / Q000662:veterinary*; D015925:Cryopreservation / Q000662:veterinary*; D004285:Dogs; D006358:Hot Temperature; D008297:Male; D055101:Semen Analysis / Q000379:methods / Q000662:veterinary; D012662:Semen Preservation / Q000662:veterinary*; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology* / Q000648:ultrastructure