Veterinary Andrology and Reproductive Techniques

PROCEDURE AND INTERPRETATION OF RESULT

(Materials required, Procedure and Result)

Materials required

• Fresh semen
• Egg yolk citrate diluent
• 5 ml test tube
• Incubator
• Methylene blue solution (50 mg of methylene blue in 100 ml of 2.9 per cent sodium citrate buffer)
• Liquid paraffin
• Water bath

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Procedure

• Take 0.2 ml of fresh semen and 0.8 ml of egg yolk citrate diluent in a sterile 5 ml test tube.
• Add 0.1 ml methylene blue solution and mix the contents.
• Place 1 cm layer of liquid paraffin.

Liquid paraffin is layered to create anerobic environment.

• Keep the test tube in a water bath of 46.5⁰C.
• Observe the time taken to change in colour from blue to colourless.

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Result

Based on the time taken to change the colour, the semen sample is classified as follows.

S.No.	Time interval	Classification
1	3 - 5 min	Good
2	9 min	Average
3	> 9 min	Poor

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