*4.5. Statistical Analysis*

To test the effects of temperature, pH, and ionic strength on antibacterial activity of oocyte lysate, one-way permutational multivariate analyses of variance (PERMANOVA) [80] were performed based on Euclidean distances on untransformed data, using 9999 random permutations of the appropriate units [81], following three different designs with one factor, separately: temperature (Te, as fixed factor with four levels); pH (pH, as fixed factor with five levels); ionic strength (IS, as fixed factor with three levels). When significant differences were found (*p* < 0.05), post hoc pairwise tests were carried out to ascertain the consistency of the differences among several different conditions tested. Because of the restricted number of unique permutations in the pairwise tests, p values were obtained from Monte Carlo samplings. The analyses were performed using the software PRIMER v. 6 [82].
