In vitro fertilization with cryopreserved spermatozoa in small pieces of gonad of the scallop Argopecten purpuratus (Lamarck, 1819)

• Published in: Cryobiology 2016-10, Vol.73 (2), p.216-220
• Main Authors: Dupré, Enrique, Covarrubias, Alejandra, Goldstein, Merari, Guerrero, Alicia, Rojas, Herman
• Format: Article
• Language: English
• Subjects: Animals; Argopecten purpuratus; Cryoadditive; Cryopreservation; Cryopreservation - methods; Cryoprotective Agents - pharmacology; Dimethyl Sulfoxide - pharmacology; DMSO; Female; Fertilization in Vitro - methods; Freezing; Freezing rates; Gonads; Male; Me2SO; Mollusks; Pectinidae; Semen Preservation - methods; Spermatozoa - drug effects
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1016/j.cryobiol.2016.07.008

The aim of this work was to evaluate the fertilization rates through the first cleavage, obtained with fresh oocytes inseminated with sperm cryopreserved at different freezing rates (−8.8,−10 and −12 °C/min) and at two thawing rates, using cryoprotectant solution (Me2SO, at 1 M, 2 M, 3 M, 4 M and 5 M with or without egg yolk and sucrose). Sperm contained in small pieces of male gonad were frozen at the three freezing rates, stored in liquid nitrogen and later the samples were thawed at two rates by immersing the samples in water at 50 °C (rapid) or 30 °C. Control fertilization rates ranged from 69.2 ± 2.8%–45.5% ± 1.6%. To determine the best concentration of the cryoprotectant (between 1 M and 5 M), in a first step, a freezing of −15 °C/min and a rapid thawing was used. Fertilization rates ranged between 9.6 ± 2.5% and 34.6± 12.2% and the highest percentages of fertilized oocytes (34.6%) was obtained with 3 M concentration with cryoaditive. The second step, using 3 M Me2SO with cryoadditive, determined that the freezing rate −8.8 °C/min produces the best result 29 ± 2.9% of fertilized oocytes corresponding to 59.2 ± 9.1% compared to controls. Although there were no significant differences among the different freezing rates, the fertilization rate tended to be higher with a lower freezing rate. Comparing the results of the present study, which used a cryoprotective solution composed of Me2SO and a cryoadditive, to those of other studies that used Me2SO without cryoadditives, suggests that the addition of a cryoadditive to the cryoprotectant Me2SO improves the fertilizing capacity of the sperm of Argopecten purpuratus after being cryopreserved.