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Home / Equine Research Bank / Equine Vet J / Post-cooling semen processing and sperm re-suspension as an ...
Equine veterinary journal2024; doi: 10.1111/evj.14085

Post-cooling semen processing and sperm re-suspension as an alternative method to circumvent poor semen cooling in stallions.

Abstract: Artificial insemination with cooled-shipped semen is the primary method used in the equine breeding industry; yet, sperm quality and fertility can be suboptimal for some stallions when standard techniques are used. Therefore, there is a critical need to develop alternative approaches for these stallions. Objective: To assess sperm quality parameters and fertility of cooled-stored stallion semen processed by SpermFilter® or centrifugation and resuspended in three extenders. Methods: Controlled and field study. Methods: In Experiment 1, semen was collected from 21 stallions classified as having good ('Good-coolers', n = 8) or poor ('Bad-coolers', n = 13) semen cooling. The semen was extended at 30 million spermatozoa/mL in a skimmed milk-based (SM) diluent, and refrigerated for 24 h. Then, the cooled-stored semen was processed through SpermFilter® or centrifugation, and the resulting sperm pellets were resuspended in SM, SM containing pentoxifylline (SM-P), or an egg yolk-based (EY) extender. Unprocessed cooled-stored semen served as control. Sperm motility parameters, plasma membrane integrity (PMI), and mitochondrial membrane potential (HMMP) were assessed in cooled-semen pre- and post-processing. Experiment 2, cooled semen from 9 stallions classified as Bad-coolers was used to inseminate 18 embryo donor mares at 66 cycles (Unprocessed, n = 22; SpermFilter®/SM-P, n = 16; or SpermFilter®/EY, n = 28). Data were analysed with a mixed model and Tukey's as posthoc, and logistic regression. Results: Processed semen resuspended in EY had superior sperm motility compared to unprocessed, SM and SM-P (p  0.05). Pellet resuspension with EY and SM-P improved the HMMP of Bad-cooler stallions (p = 0.0010). Semen processed by SpermFilter® had superior PMI to centrifuged semen (p < 0.0001). Mares inseminated with SpermFilter®/SM-P (50%, 8/16) or SpermFilter®/-EY (68%, 9/28) had higher pregnancy rates than mares bred with unprocessed semen (14%, 3/22) (p < 0.001). Conclusions: Low number of mares in the fertility trial. Conclusions: Sperm quality and fertility of Bad-cooler stallions can be enhanced by SpermFilter® and pellet resuspension with either EY or SM-P.
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Publication Date: 2024-04-03 PubMed ID: 38567428DOI: 10.1111/evj.14085Google Scholar: Lookup
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Summary

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The research explores the viability of post-cooling semen processing and sperm re-suspension methods to improve fertility rates in stallions known to have poor semen cooling, a common issue in the equine breeding industry. The studied techniques reportedly provided significant improvements in sperm quality and mare pregnancy rates.

Methods Investigated

  • The study involved 21 stallions, divided into two groups – those with good semen cooling (Good-coolers) and those with poor semen cooling (Bad-coolers).
  • The stallions’ semen was extended to 30 million spermatozoa per milliliter, with a skimmed milk-based (SM) diluent, then refrigerated for 24 hours.
  • The semen then went through a SpermFilter® or was subjected to centrifugation. The filtered or centrifuged semen was then resuspended in SM, SM with an added drug called pentoxifylline (SM-P), or an egg yolk-based (EY) extender.
  • The researchers compared these samples to unprocessed semen, assuming them as control samples.
  • Sperm motility parameters, plasma membrane integrity (PMI), and high mitochondrial membrane potential (HMMP) were the sperm quality parameters assessed both pre- and post-processing.
  • A second part of the study involved breeding 18 embryo donor mares with the cooled semen of nine Bad-coolers. Different processing methods were tested across 66 breeding cycles to assess their impact on actual pregnancy rates.

Results and Conclusions

  • Cooled semen post-processing and resuspension with EY displayed better sperm motility than unprocessed semen, SM, and SM-P.
  • Semen processed with a SpermFilter® and resuspended in SM-P was roughly equivalent to EY in results.
  • The HMMP of Bad-coolers improved significantly with EY and SM-P resuspension, showing better sperm quality.
  • The PMI was also superior in semen processed by the SpermFilter® compared to centrifuged semen.
  • Finally, the fertility tests showed that mares bred with semen processed with SpermFilter® and resuspended in either SM-P or EY had considerably higher pregnancy rates than those bred with unprocessed semen.
  • Despite the limited number of mares used for fertility trials, the study concluded that SpermFilter® combined with EY or SM-P resuspension could increase sperm quality and fertility in stallions with poor semen cooling capabilities.

Cite This Article

APA
(2024). Post-cooling semen processing and sperm re-suspension as an alternative method to circumvent poor semen cooling in stallions. Equine Vet J. https://doi.org/10.1111/evj.14085

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English

Researcher Affiliations

Grant Funding

  • 2017/13883-9 / Fundau00e7u00e3o de Amparo u00e0 Pesquisa do Estado de Su00e3o Paulo

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