Tapirus bairdii (Spermatozoa (sperm))

(Cryopreservation of Baird’s Tapir (Tapirus bairdii) Sperm)

Intro/Description:

This protocol is reported in an article describing ejaculate traits of the endangered Baird’s Tapir (Tapirus bairdii). The authors note that while this protocol is generally successful, it does lead to a statistically significant decrease in sperm motility and acrosomal integrity.

Materials:

• Tapirus bairdii ejaculate
• 50 ml centrifuge tubes
• Centrifuge
• Botu-Crio (CryoVital, Grandau, Germany)
• Egg yolk
• Methylformamide
• Glycerol
• 0.5 ml plastic straws
• Liquid nitrogen
• Cryovials

Protocol:

1. Place collected Tapirus bairdii ejaculate in a sterile 50 ml centrifuge tube.
2. Centrifuge at 500 x g for 15 minutes. Discard supernatant.
3. Resuspend sperm pellets in Botu-Crio with 20% (v/v) egg yolk, 1% (v/v) glycerol and 4% (v/v) methylformamide to a dilution of 250 x 106 motile sperm/mL.
4. Package sperm suspension into 0.5 ml plastic straws, then heat seal the straws.
5. Cool the straws for 20 min at 5°C.
6. Freeze the straws by placing them in liquid nitrogen vapor (4 cm above the liquid) for 15 minutes.
7. Plunge the straws into liquid nitrogen. They are now prepared for long-term storage.
8. To thaw, place frozen straws at 46°C for 20 seconds, and empty the straw contents into a sterile tube.

References:

Pukazhenthi BS, Togna GD, Padilla L, Smith D, Sanchez C, Pelican K, Sanjur OI. Ejaculate traits and sperm cryopreservation in the endangered Baird's tapir (Tapirus bairdii). J Androl. 2011 May-Jun;32(3):260-70.

Added by: Carlton Hoyt on July 25, 2012

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