title: Effect of cryopreservation and single layer centrifugation on canine sperm DNA fragmentation assessed by the sperm chromatin dispersion test.
abstract: The aims of this study were: 1) to assess the effect of freezing and thawing on dog sperm DNA fragmentation index (sDFI) using the sperm chromatin dispersion test (SCDt); and 2) to determine whether or not the sperm selection by single layer centrifugation (SLC) using Androcoll-C improves sperm DNA longevity in SLC-selected frozen-thawed dog semen samples. Semen samples were collected from 4 dogs using digital manipulation. After collection, ejaculates were pooled and cryopreserved following a standard protocol. Sperm motility and morphology were assessed before freezing and after thawing as a control for the cryopreservation method used. In experiment 1, sDFI was analyzed immediately before freezing and after thawing (baseline values), showing no significant differences between fresh and frozen-thawed semen samples. In experiment 2, frozen-thawed semen samples were processed or not by SLC using Androcoll-C and longevity of DNA were assessed in terms of sDFI after 24h of in vitro incubation at physiological temperature (38°C). The results showed low values of sDFI in SLC-selected semen in comparison to unselected samples. In conclusion, no effect of cryopreservation was observed on baseline values of dog sperm DNA fragmentation. Additionally, SLC-selection using Androcoll-C improved longevity of frozen-thawed sperm DNA assessed by the SCDt.
authors.affiliation: Animal Reproduction Group, Department of Animal Medicine and Surgery, University of Cordoba, Cordoba, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Urbano
authors.forename: J
authors.initials: J
authors.lastname: Dorado
authors.forename: I
authors.initials: I
authors.lastname: Ortiz
authors.forename: J M
authors.initials: JM
authors.lastname: Morrell
authors.forename: S
authors.initials: S
authors.lastname: Demyda-Peyrás
authors.forename: M J
authors.initials: MJ
authors.lastname: Gálvez
authors.forename: L
authors.initials: L
authors.lastname: Alcaraz
authors.forename: L
authors.initials: L
authors.lastname: Ramírez
authors.forename: M
authors.initials: M
authors.lastname: Hidalgo
mesh_terms: D000818:Animals; D002498:Centrifugation / Q000379:methods / Q000662:veterinary; D002843:Chromatin / Q000378:metabolism*; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary; D053938:DNA Fragmentation; D004285:Dogs / Q000235:genetics / Q000378:metabolism; D005615:Freezing / Q000009:adverse effects; D008297:Male; D055101:Semen Analysis / Q000379:methods* / Q000662:veterinary; D012662:Semen Preservation / Q000009:adverse effects / Q000379:methods / Q000662:veterinary; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*
chemical_list: D002843:Chromatin
keywords: Androcoll-C; Cryopreservation; DNA fragmentation; Dog; SCDt