DMSO associado com açúcares na criopreservação espermática de Pacu

Abstract

The technique of sperm cryopreservation is simple and allows the creation of germplasm banks, allowing the conservation of genetic diversity and replacement of endangered species. However, during the cryopreservation process the frozen cells can be damaged by cryoprotectant toxicity by ice crystals, resulting in damage to DNA, mitochondria, membrane and interfering with sperm kinetics. The objective of this work was to evaluate the effect of the association of 10% DMSO on trehalose, raffinose, sucrose and lactose at 50 mM, 100 mM and 150 mM concentrations on the sperm cells of Piaractus mesopotamicus. Sperm were collected from the animals through abdominal massage, being hormonally induced with carp pituitary extract 12h before extrusion. Then the samples were diluted in Beltsville Thawing Solution (BTS) containing the penetrant cryoprotectant dimethylsulfoxide (DSMO) at 10% concentration (control) and in the other treatments were added the different concentrations of sugars. After 60 days, cell movement analysis (total and progressive motility, DAP, DCL, DSL, VAP, VCL, VSL, STR, LIN, WOB, ALH and BCF) were performed by Computer Assisted Semen Analysis (CASA) And analysis of cell structures: DNA integrity, membrane integrity, mitochondrial functionality, membrane fluidity and reactive oxygen species (ROS), through flow cytometry. The Shapiro - Wilk normality test was performed, with subsequent analysis of variance using the Kriskal Wallis test, because the data did not behave in a parametric manner. The evaluations of total motility and motility period were better when associated with 100 mM raffinose at 10% DMSO (P <0.05). The lateral displacement of the head (ALH) obtained a good result when using the association of 10% DMSO with 100 mM lactose, 150 mM sucrose and 150 mM raffinose, showing better (P <0.05) than the treatment without Addition of lactose (0 mM). In conclusion, considering the kinetic parameters essential for sperm cryopreservation, the results of the motility and total motility period evaluations presented their best rates in the treatment containing 100 mM raffinose associated with 10% DMSO.