Comparison of two dilution rates on canine semen quality after cryopreservation in a coconut water extender

The objective of the present study was to evaluate the effect of sperm dilution (one part semen:one part extender or at 200 × 106 spermatozoa/mL) using a coconut water extender on the post-thaw sperm quality. Twelve ejaculates were collected from six dogs. Semen was divided into two aliquots, one for dilution one part semen:one part extender (group 1) and another for a concentration of 200 × 106 spermatozoa/mL (group 2). Semen was initially extended at 37 °C at a proportion of one part semen:half part extender (1:12) for group 1 (A-fraction). For group 2, the volume for a concentration of 200 × 106 spermatozoa/mL was calculated and a half of this volume was used for the initial dilution (A-fraction, 37 °C). Coconut water extender containing 20% egg yolk was used for this initial dilution in both groups. After dilution, the semen was cooled for 40 min in a thermal box (15 °C) and for 30 min in a refrigerator. The other half of the extender (B-fraction) containing egg yolk and glycerol (12%) was added to semen in both groups. Subsequently, the final concentration of glycerol in the extender was 6%. Ejaculates were frozen in 0.25 mL straws 5 cm above the surface of liquid nitrogen and stored at −196 °C. After 1 week, straws were thawed at 37 °C for 1 min and the microscopic criteria were evaluated. The dilution method had no influence on sperm motility, vigor and normal spermatozoa (71.4 compared with 67.7%). There was no effect of dog, ejaculate within male on post-thaw semen quality. Moreover, there was not a male × treatment interaction. Both treatments were efficient in preserving sperm quality.