Selection of frozen–thawed stallion semen by microfluidic technology

Vigolo, Veronica; Gautier, Camille; Falomo, Maria Elena; Aurich, Christine

doi:10.1111/rda.14305

Cited by 5 publications

(1 citation statement)

References 24 publications

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“…13 Therefore, to optimise the longevity of frozenthawed spermatozoa, it is desirable to select a high proportion of viable spermatozoa. New microfluidic sperm sorting methods used in in vitro embryo production systems have been shown to select subpopulations of frozen-thawed spermatozoa with high motility, 14 normal morphology, viability, membrane integrity and DNA integrity without the need for centrifugation as required for other sperm separation techniques like density gradient separation. 15,16 Centrifugation of spermatozoa has been shown to increase the level of reactive oxygen species in the sample, which is detrimental to sperm DNA integrity and fertility 17 and so the ability to eliminate a centrifugation step may be beneficial.…”

Section: Introductionmentioning

confidence: 99%

A simplified fixed‐time insemination protocol using frozen–thawed stallion spermatozoa stored at 17°C for up to 24 h before insemination

Morris,

Harteveld,

Gibb

2024

Equine Veterinary Journal

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BackgroundInsemination of mares with frozen–thawed spermatozoa requires intensive management and results in 40%–60% per cycle pregnancy rates.ObjectivesTo determine if satisfactory fertility is possible for frozen–thawed semen after processing it through a microfluidic device, followed by storage at 17°C for up to 24 h before fixed‐time insemination.Study designUncontrolled field trials.MethodsA pilot study evaluated the motility of frozen–thawed spermatozoa after centrifugation and storage (17°C) in two different media for up to 48 h. Subsequently, the motility of frozen–thawed semen processed through a microfluidic device, resuspended in two different media during storage (17°C) for up to 24 h was evaluated. The fertility of frozen–thawed spermatozoa, after microfluidic sorting and storage at 17°C for up to 24 h, was evaluated after fixed‐time insemination in a commercial embryo programme. Experiment 1: Frozen–thawed spermatozoa (N = 5 stallions) were centrifuged and resuspended in Botusemen Gold™ or SpermSafe™ and stored (17°C) for up to 48 h. Sperm motility was evaluated by CASA at 0, 6, 24 and 48 h. Experiment 2: Frozen–thawed spermatozoa (N = 4 stallions) underwent microfluidic sorting and storage (17°C) for up to 24 h in both media. Sperm concentration and motility were evaluated at 0, 16 and 24 h. Experiment 3: Fertility of frozen–thawed spermatozoa (N = 3 stallions) was evaluated after insemination of 42 mare cycles at 6, 16 and 24 h after thawing, microfluidic sorting and storage before fixed‐time insemination.ResultsThe stallion significantly influenced sperm motility, but there was no effect of media on motility parameters. Storage time significantly affected sperm motility after centrifugation but not after microfluidic sorting. Storage time had no effect on the overall embryo recovery rate (52%, n = 42).Main limitationsField trial with small mare numbers and no control at time = 0 h.ConclusionsFixed‐time insemination of frozen–thawed spermatozoa after microfluidic sorting and storage at 17°C for up to 24 h produced satisfactory embryo recovery rates.

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Section: Introductionmentioning

confidence: 99%

A simplified fixed‐time insemination protocol using frozen–thawed stallion spermatozoa stored at 17°C for up to 24 h before insemination

Morris,

Harteveld,

Gibb

2024

Equine Veterinary Journal

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Protamine 2 and phospholipase C zeta 1 are possible biomarkers for the diagnosis of male subfertility in frozen-thawed stallion semen

Vigolo,

Gautier,

Ertl

et al. 2024

Theriogenology

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Freezing Stallion Semen—What Do We Need to Focus on for the Future?

Al-Kass,

Morrell

2024

Veterinary Sciences

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Artificial insemination (AI) is used frequently in the breeding of sport horses, apart from Thoroughbreds. Most AIs are carried out with cooled semen rather than frozen semen because of the difficulties in identifying a protocol that is suitable for freezing most ejaculates and the necessity to inseminate close to ovulation because of the short life of the thawed spermatozoa. More widespread use of frozen semen would improve biosecurity, allow greater choice of stallions, and offer more flexibility when managing deliveries of semen to the stud. It would even decrease the amount of antibiotics used in semen extenders, since the volume of frozen semen is smaller than when cooled semen is inseminated. However, there is considerable variability in the cryosurvival of spermatozoa from different stallions, leading to the classification of stallions as good or bad freezers. Improvements could be made at the level of stallion nutrition, the semen collection regimen, the extender, the removal of seminal plasma, and the cooling protocol, among others. Stallion sperm membranes are highly susceptible to lipid peroxidation, but research on antioxidants has failed to identify an additive that would benefit all stallions. In the future, biomarkers for sperm freezability could be used as an aid in identifying suitable ejaculates for cryopreservation.

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Selection of frozen–thawed stallion semen by microfluidic technology

Cited by 5 publications

References 24 publications

A simplified fixed‐time insemination protocol using frozen–thawed stallion spermatozoa stored at 17°C for up to 24 h before insemination

A simplified fixed‐time insemination protocol using frozen–thawed stallion spermatozoa stored at 17°C for up to 24 h before insemination

Protamine 2 and phospholipase C zeta 1 are possible biomarkers for the diagnosis of male subfertility in frozen-thawed stallion semen

Freezing Stallion Semen—What Do We Need to Focus on for the Future?

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