Development of Two Stages Coc’s Semen Extender for Room Temperature Storage at Laboratory

Abstract

A simple cock’s semen dilutor (two-stage) with minimum ingredients was developed in the present study. The stage-1dilutor was aimed to reduce the motility and metabolism during storage and the stage-2 dilutor was to be added just prior to insemination to revive the motility to facilitate the entry of spermatozoa into sperm host gland. Initially, the semen was diluted 1 : 1 with the stage-1 dilutor having lower pH that reduced the motility and metabolism. Subsequently, after 24h, the diluted semen was mixed slowly with another dilutor stage-2 at 1 : 2 ratios to revive the motility of the spermatozoa to normal level. Phosphate buffer was the base for both storage (stage-1) and recovery (stage-2) dilutors with a pH of 6.5 and 7.5, respectively. In stage-1 dilutor sodium glutamate to minimize metabolism of spermatozoa and antibiotics to check bacterial growth were used whereas these were absent in stage-2 dilutor. Glucose was added as energy source in the dilutor of both stages. The osmolarity of stage-1 and stage-2 dilutors was 346 and 327Osm/kg H₂0, respectively. This dilutor can be used to store the cock’s semen at room temperature (17-21°C) for 24h with sustained motility. This new approach for two-stage dilutor of cock’s semen storage at room temperature for extended period could be a useful A.I. tool under farm condition.