Sas package v 9

Acrosome integrity and embryo development data were handled in GraphPad Prism for Windows (Version 6.01, GraphPad software, CA, USA) using Fisher’s exact test and One-way Anova non-parametric test (for acrosome integrity and embryo development respectively). Tests were based on more than five replicates per experiment. Data are reported as means ± standard error mean (SEM).
TGA analysis were run in triplicate and the results reported as average.
Data obtained from SCSA assessment were analysed using the SAS package v 9.4 (SAS Institute Inc., Cary, NC, USA). The General Linear Model procedure (PROC GLM) was used to evaluate the effect of the freeze-dry protocols on the SCSA sperm quality parameters. The model included the fixed effect of the ram. Results are given as adjusted least squares means ± standard error means (LSM ± SEM). A PROC CORR was applied to calculate Spearman’s correlation coefficients within freeze-dried sperm quality and embryo development variables.
For all the tests the level of significance was set at P < 0.05.

Data obtained from CASA and flow cytometry measurements were analyzed using the SAS™ package v 9.4 (SAS Institute Inc., Cary, NC, USA). The General Linear Model procedure (PROC GLM) was used to analyze the effect of technical replicates on semen quality parameters in the two fractions. The model included as fixed effects the bull and the replicate nested in the sperm fractions (HM and LM).
A mixed model procedure (PROC MIXED) was used to perform analysis on sperm quality parameters in order to evaluate the efficiency of the sperm separation into the HM and LM sperm fractions. The mixed model included the fixed effect of the sperm fraction (HM and LM), and bull as random. Results are given as adjusted least squares means ± standard error means (LSM ± SEM).