title: Sperm yield after single layer centrifugation with Androcoll-E is related to the potential fertility of the original ejaculate.
abstract: Many attempts have been made to identify laboratory tests that are predictive of sperm fertility, both to improve the quality of stallion semen doses for artificial insemination (AI) and to identify potential breeding sires if no fertility data are available. Sperm quality at the stud is mostly evaluated by assessing subjective motility, although this parameter can be poorly indicative of fertility. Sperm morphology and chromatin integrity in Swedish stallions are correlated to pregnancy rate after AI. Because single layer centrifugation (SLC) selects for spermatozoa with normal morphology and good chromatin, retrospective analysis was carried out to investigate whether sperm yield after SLC is linked to potential fertility. Commercial semen doses for AI from 24 stallions (five stallions with four ejaculates each, 19 stallions with three ejaculates each; n = 77) obtained during the breeding season were cooled, and sent overnight to the Swedish University of Agricultural Sciences in an insulated box for evaluation, with other doses being sent to studs for commercial AI. On arrival at Swedish University of Agricultural Sciences, the semen was used for SLC and also for evaluation of sperm motility, membrane integrity, chromatin integrity, and morphology. The seasonal pregnancy rates for each stallion were available. The yield of progressively motile spermatozoa after SLC (calculated as a proportion of the initial load) was found to be highly correlated with pregnancy rate (r = 0.75; P < 0.001). Chromatin damage was highly negatively correlated with pregnancy rate (r = -0.69; P < 0.001). Pregnancy rate was also correlated with membrane integrity (r = 0.58; P < 0.01), progressive motility (r = 0.63; P < 0.01), and normal morphology (r = 0.45; P < 0.05). In conclusion, these preliminary results show that sperm yield after SLC is related to the potential fertility of the original ejaculate, and could be an alternative indicator of stallion fertility if breeding data are not available. Single layer centrifugation is fast (30 minutes) and does not require expensive equipment, whereas other assays require a flow cytometer and/or specialist skills. An additional option could be to transport semen doses to a laboratory for SLC if the stud personnel do not want to perform the procedure themselves.
authors.affiliation: Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden. Electronic address: jane.morrell@kv.slu.se.
authors.forename: J M
authors.initials: JM
authors.lastname: Morrell
authors.affiliation: Flyinge AB, Flyinge Kungsgård, Sweden.
authors.forename: G
authors.initials: G
authors.lastname: Stuhtmann
authors.affiliation: Flyinge AB, Flyinge Kungsgård, Sweden.
authors.forename: S
authors.initials: S
authors.lastname: Meurling
authors.affiliation: Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Lundgren
authors.affiliation: Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
authors.forename: C
authors.initials: C
authors.lastname: Winblad
authors.affiliation: Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
authors.forename: B
authors.initials: B
authors.lastname: Macias Garcia
authors.affiliation: Anatomy Physiology and Biochemistry, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Johannisson
mesh_terms: D000818:Animals; D001947:Breeding; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002498:Centrifugation / Q000379:methods / Q000662:veterinary*; D005260:Female; D005298:Fertility; D006736:Horses; D007315:Insemination, Artificial / Q000662:veterinary; D008297:Male; D011247:Pregnancy; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000002:abnormalities / Q000166:cytology / Q000502:physiology*; D013548:Sweden
keywords: Androcoll-E; Colloid centrifugation; Seasonal pregnancy rate; Sperm quality; Yield of motile spermatozoa