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Semen Freezing

Why Freeze?

  • To enable you to market your stallion overseas or more widely in the UK.
  • To have a bank of semen stored in case of accident or injury.
  • Join the growing number of owners wishing to use frozen semen to avoid distraction to the competing stallion.
  • For all these reasons we are here to offer a professional service.

Collection of Semen

Semen is collected using one of several models of an artificial vagina (AV). The AV is lubricated thoroughly using a non-spermicidal sterile gel. The semen is collected in a sterile bottle designed to separate the sperm rich fraction from the often-copious gel fraction. A typical stallion will produce between 30-70ml sperm rich semen. The semen is quickly diluted with a transport medium, which is a cocktail of a series of chemicals, which maintains the viability of sperm for up to 7 hours from the collection of sperm. This remarkable solution is capable of holding the semen at room temperature, and therefore the necessity of chilling and incubating is removed. The sperm are checked for motility at this stage.

The laboratory at Stallion AI Services is equipped with the latest high tech apparatus for the correct processing, freezing and evaluation of semen from your valuable stallion.

Centrifugation of the ejaculate

Sperm are then spun down on to a liquid cushion to prevent damage during centrifugation; this is the process of reducing the volume of semen resulting in an increased density of sperm. The semen is centrifuged at 1000 times the force of gravity for 25 minutes. Once spun down, the band of cells suspended on the liquid cushion can be harvested ready for the next stage of processing.

Photometric equipment is used to accurately calculate the exact number of sperm in the ejaculate. When this has been established the semen can be diluted with a freezing extender to the correct concentration for insemination. 6 hundred million sperm will eventually be used to inseminate into the mare.

Filling and Printing of straws

The diluted semen will be packaged into 0.5 ml plastic straws. Each straw is printed with information relating to the stallion. This information includes the ministry licence number, the stallion’s name, date of semen collection and the stallion’s registration number. The printed straws are then placed in a machine that will fill and seal each one. The machine will automatically fill each straw with exactly 0.5ml of semen. Each straw contains 75 million sperm cells and 6 of these straws will make up the single insemination dose.

Freezing

The straws containing the semen are then placed into the digit cool semen freezer. This Cryogenic machine uses liquid nitrogen to freeze the semen. A computer linked to the freezing machine is pre-programmed to cool and freeze the semen at a specific rate down to a chilly –140C. Once at this temperature the straws can be removed from the digit cool and plunged into liquid nitrogen at –196C. At this temperature the sperm are in suspended animation. The sperm will remain viable indefinitely at this temperature and will not degrade or perish. The semen can be used to inseminate mares many years after it was collected from the stallion. Random straws are removed for post thaw quality control.

Evaluation


After freezing, 2 straws from each ejaculate are tested by one of the centre’s experienced technicians. They are thawed at 37C for 35 seconds and pooled in a tube. The motility is assessed visually and must have at least 35% progressive motility. The semen is then stained with propidium iodide (PI) and carboxyflourescein diacetate (CFD). The PI stains the dead sperm cells red and the CFD stains live cells green. After counting 200 cells, the live percentage is calculated using a formula. A morphology assessment is done by looking at 100 live cells and noting the number of normal ones. The number of live, normal sperm post-thaw in each dose can then be calculated to form a clear objective view of the end product, which is to be inseminated into the mare. To pass the post-thaw quality tests at least 260 million cells must be live and normal, of the 800 million in each dose of 4mls.

Storage

After semen has successfully passed the post thaw evaluation assessments, the semen is stored in our new, purpose built storage centre on site.

Storage & Distribution

Attached documents