Abstract
Artificial insemination with doses containing low-sperm numbers has been utilized to optimize the use of elite bulls. Hen egg yolk is widely used as a cryoprotective agent in semen freezing extender protecting the spermatozoa. Its action is due to the presence of low-density lipoproteins (LDL) in the hen egg yolk. The objectives of the present study were to evaluate the effects of the semen dilution to low-sperm number/dose on sperm motility and integrity of sperm plasma membrane in the cryopreservation process, using two commercial extenders (Triladyl ®, Bioxcell ®) and LDL extender prepared in our laboratory, 97% purity. Fifteen ejaculates were collected from five fertile crossbred bulls ( Bos taurus × Bos indicus). After collection, sperm motility was examined by Computer-Assisted Semen Analysis (Hamilton Thorne), morphological sperm characteristics were evaluated by differential interference microscopy and the integrity of plasma membranes was determined using the hypo-osmotic swelling test. The semen was subsequently divided into three aliquots and diluted with the three extenders into 120 × 10 6, 60 × 10 6 and 20 × 10 6 sperm/mL, corresponding to 30 × 10 6, 15 × 10 6 and 5 × 10 6 sperm/dose, respectively. This study revealed that LDL extender was more effective in preservation of motility and integrity of the plasma membrane of spermatozoa than Bioxcell ® and Triladyl ® ( p < 0.05), but no significant difference was observed between Triladyl ® and Bioxcell ®. Therefore we can conclude that LDL extender could be used instead of Triladyl ® or Bioxcell ® at low semen concentration per dose for elite bulls, it also could be envisaged for the industry of sex-stored semen.
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