Field fertility of liquid stored and cryopreserved flow cytometrically sex-sorted stallion sperm.
Abstract: The fertility of sex-sorted, cryopreserved stallion sperm must be improved for the sex-sorting technology to be applied commercially. Objective: To optimise the conditions used to liquid store stallion sperm prior to sex-sorting and assess the fertility of sperm following sex-sorting and cryopreservation. Methods: Both in vitro experiment and randomised controlled trial in healthy, client-owned mares. Methods: Stallion ejaculates (n = 9) were diluted in either a skimmed milk (KMT) or BSA (I-BSA) based media to 25 × 106 sperm/ml directly (+SP25) or washed to remove seminal plasma and diluted to 25 or 111 × 106 sperm/ml (-SP25 and -SP111). Sperm were stored for 18 h at 10 to 15°C and -SP25 and +SP25 treatments were centrifuged and resuspended to 111 × 106 sperm/ml. Sperm were incubated under H33342 staining conditions and motility, viability and acrosome integrity assessed. Semen was collected from stallions (n = 4), liquid stored at 10-15°C for up to 5 h and sperm either cryopreserved directly, sex-sorted and cryopreserved, or sex-sorted and returned to liquid storage until insemination. Low-dose hysteroscopic insemination was performed in 23 mares randomly allocated to the semen preparation group and pregnancy determined following embryo flushing on Day 9 after ovulation, or via transrectal ultrasonography on Day 14 after ovulation. Results: Skimmed milk was superior to I-BSA in maintaining motility, viability and acrosome integrity. Seminal plasma removal did not affect the parameters measured at the concentrations examined. Conception rates did not differ significantly between the groups, although a high incidence of pregnancy loss was observed in both the cryopreserved groups. Conclusions: While the conception rates achieved are among the highest yet reported for sex-sorted, cryopreserved stallion sperm, the high incidence of pregnancy loss suggests that the development of the resulting embryos was significantly impaired by the sperm processing treatments.
© 2016 EVJ Ltd.
Publication Date: 2016-05-22 PubMed ID: 27060979DOI: 10.1111/evj.12583Google Scholar: Lookup
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- Journal Article
Summary
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The research study focuses on improving the fertility of sex-sorted, cryopreserved stallion sperm for commercial use. This involved optimizing liquid storage conditions of stallion sperm and assessing their fertility post sex-sorting and cryopreservation.
Methods
- Both in vitro experiments and a randomised controlled trial involving healthy, client-owned mares were conducted.
- The collected stallion ejaculates (n = 9) were diluted in either a skimmed milk (KMT) or BSA (I-BSA) based media to certain sperm concentrations, both with and without washing to remove seminal plasma.
- The sperm samples were stored at temperatures between 10 – 15°C for 18 hours with certain treatments being centrifuged and resuspended to a specific sperm concentration.
- These stored sperm samples were incubated under H33342 staining conditions and evaluated for motility, viability and acrosome integrity.
- For the trial, semen was collected from stallions (n = 4), liquid stored at similar temperatures for up to 5 hours and sperm either directly cryopreserved, sex-sorted and cryopreserved, or sex-sorted and returned to liquid storage until insemination.
- Low-dose hysteroscopic insemination was performed in 23 mares and pregnancy was determined following embryo flushing on Day 9 post ovulation, or via transrectal ultrasonography on Day 14 post ovulation.
Results
- It was found that skimmed milk was more effective than I-BSA in maintaining motility, viability and acrosome integrity of the sperm.
- Removal of seminal plasma was found to have no significant effect on the parameters measured.
- While conception rates did not significantly differ between groups, a high incidence of pregnancy loss was observed in both the groups where cryopreservation was involved.
Conclusions
- Although the conception rates achieved with cryopreserved, sex-sorted stallion sperm were the highest recorded yet, the high incidence of pregnancy loss suggests adverse effects on embryo development by the sperm processing treatments.
- Improvements in these sperm processing treatments are therefore needed, especially for successful commercial application of sex-sorting technology in stallion sperm.
Cite This Article
APA
Gibb Z, Grupen CG, Maxwell WM, Morris LH.
(2016).
Field fertility of liquid stored and cryopreserved flow cytometrically sex-sorted stallion sperm.
Equine Vet J, 49(2), 160-166.
https://doi.org/10.1111/evj.12583 Publication
Researcher Affiliations
- Faculty of Veterinary Science, The University of Sydney, New South Wales, Australia.
- Faculty of Veterinary Science, The University of Sydney, New South Wales, Australia.
- Faculty of Veterinary Science, The University of Sydney, New South Wales, Australia.
- EquiBreed NZ Limited, Te Awamutu, New Zealand.
MeSH Terms
- Acrosome / physiology
- Animals
- Cell Survival
- Cryopreservation / veterinary
- Female
- Horses / physiology
- Insemination, Artificial / veterinary
- Male
- Pregnancy
- Semen Preservation / veterinary
- Sex Preselection / veterinary
- Sperm Motility
Citations
This article has been cited 1 times.- Al-Khaldi K, Yimer N, Sadiq MB, Firdaus Jesse Bin Abdullah F, Salam Babji A, Al-Bulushi S. Edible bird's nest supplementation in chilled and cryopreserved Arabian stallion semen. Saudi J Biol Sci 2022 Mar;29(3):1577-1584.
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