title: DNA integrity of canine spermatozoa during chill storage assessed by the sperm chromatin dispersion test using bright-field or fluorescence microscopy.
abstract: The objective of this study was to evaluate the effect of chill storage on canine sperm DNA fragmentation assessed by the sperm chromatin dispersion test using bright-field microscopy with Wright solution (sDF-B) or fluorescence microscopy with propidium iodide (sDF-F). The relationship and agreement between the results obtained with both staining methods were analyzed. The values of DNA fragmentation indexes (sDF-F and sDF-B) were compared at each time of chill storage (0, 24, 48, 72, and 96 hours). Additionally, the sperm DNA fragmentation rate (slope) was compared between the methods during chill storage. Good agreement and no significant differences between values obtained with both staining procedures were observed. Finally, the effect of chill storage for up to 96 hours was assessed on sperm motility parameters and DNA fragmentation indexes. Significant differences were found after 48 hours of chill storage, obtaining greater values of fragmented DNA. Progressive sperm motility was lower just after 96 hours of chill storage, and no effect was found in total sperm motility. In conclusion, the Sperm-Halomax kit, developed for canine semen and based on the sperm chromatin dispersion test, can be used accurately under bright-field or fluorescence microscopy to assess the sperm DNA integrity of canine semen during chill storage. The sperm DNA fragmentation index increased after 48 hours of chill storage, thereby detecting sperm damage earlier than other routine sperm parameters, such as sperm motility.
authors.affiliation: Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Córdoba, Córdoba, Spain. Electronic address: mhidalgo@uco.es.
authors.forename: M
authors.initials: M
authors.lastname: Hidalgo
authors.affiliation: Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Córdoba, Córdoba, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Urbano
authors.affiliation: Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Córdoba, Córdoba, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Ortiz
authors.affiliation: MERAGEM Research Group, Department of Genetics, Faculty of Veterinary Medicine, University of Cordoba, Cordoba, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Demyda-Peyras
authors.affiliation: Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Córdoba, Córdoba, Spain.
authors.forename: M R
authors.initials: MR
authors.lastname: Murabito
authors.affiliation: Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Córdoba, Córdoba, Spain.
authors.forename: M J
authors.initials: MJ
authors.lastname: Gálvez
authors.affiliation: Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Córdoba, Córdoba, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Dorado
mesh_terms: D000818:Animals; D002843:Chromatin; D003080:Cold Temperature; D053938:DNA Fragmentation; D004285:Dogs / Q000502:physiology*; D008297:Male; D055101:Semen Analysis / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology*
chemical_list: D002843:Chromatin
keywords: Bright field; Fluorescence; Halomax; Sperm DNA; Sperm chromatin dispersion