Fertilisation experiments were performed to compare the fertilisation rate of frozen sperm and
fresh sperm. In the first experiment, artificial fertilisation was performed by placing 1 or 2 ml of
HBSS-extended fresh MGC sperm (sperm-to-extender ratio = 1:6 v/v) and eggs (1 gram) in a Petri
dish and mixing for 2 minutes using a rooster’s tail feather and 50 ml of distilled water was then
added. The mixture was placed in a small fine net enclosure (6"x6"x6"). Aeration was provided in a
nursery concrete pond. After approximately 24 hours, the fertilisation rate was determined by
observing the fertilised eggs (somite stage) under a dissecting microscope.