Selection bovine frozen semen in Percoll® gradient on spermatic parameters

Abstract

At in vitro embryo production sperm selection techniques are used, one of them is Percoll® density-gradient. The aim with this study was to evaluate the plasmatic membrane (eosin-nigrosin), acrosomal (trypan blue-Giemsa; fluorescein isothiocyanate conjugated peanut agglutinin lecitin/propidium iodide (FITC-PNA/PI) and chromatin (toluidine blue) integrity, in frozen bovine spermatozoa. Twenty nine samples were analysed at post-thaw (PD) and post-Percoll® moments. Variable expressed in percentage were submitted to ANOVA and used Tuckey’s test for mean comparison. The association between variables was made by Pearson’s correlation. There were significant differences (p<0.05) between moments for motility, plasmatic membrane integrity and number of live spermatozoa with intact acrosome (FICT -PNA/PI), with higher values for PP. The percentage of cells with chromatin alteration did not differed between moments. In conclusion, Percoll® density-gradient was efficient to select a higher population of motile cells with integral plasmatic and acrosomal membranes, without alterations in nuclear chromatin condensation.