Veterinary Andrology and Reproductive Techniques

PROCEDURE AND INTERPRETATION OF RESULT

Procedure

• Take 0.9 ml of HOS media/distilled water in a sugar tube
• Keep the tube in 37⁰C water bath for 5 minutes to bring the temperature of media to 37⁰C
• Then add 0.1 ml of semen
• Incubate the mixture at 37⁰C water bath for 30 minutes
• Place a drop of semen in a clean, grease free glass slide and put a cover slip over it.
• Examine under phase contrast microscope to see the tail curling

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If the clarity is not there, do the following procedure

• Make a thin smear of the incubated mixture in a clean slide and dry it.
• Stain the slide with 3% Rose Bengal stain for 20 minutes
• Wash the slide under running tap water and dry it
• Examine the slide under oil immersion microscope for tail curling

• Count at least 200 spermatozoa
  • Calculate the percentage of spermatozoa with showing tail curling.
  • Samples showing higher percentage of sperms with tail curling indicate good samples

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Calculation

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• The fresh semen samples should have a minimum of 70% and above HOS reacted sperms.
• The frozen semen samples should have a minimum of 50% reacted sperm.

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