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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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.

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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

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Equine veterinary journal

ISSN: 2042-3306

NlmUniqueID: 0173320

Country: United States

Language: English

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Grant Funding

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

References

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