Prism 4.0 for windows

Data obtained were expressed as mean ± standard error of the mean (Mean ± SEM). Cosinor analysis was used to determine the t_cloacal daily rhythms of individual birds. The mean mesor (rhythm‐adjusted mean), amplitude (half the range of excursion or a measure of the extent of predictable change within a cycle) and acrophase (time of peak) values of the variables of daily rhythm were calculated for each bird and for each time series of the study period. Values were subjected to repeated‐measures one‐way analysis of variance (ANOVA model‐3) and by the cosinor procedure (Refinetti et al. 2007; Piccione et al. 2013), followed by Tukey's multiple comparison post hoc test, using GraphPad Prism 4.0 for windows (GraphPad Software, San Diego, CA) to compare the differences between the means, obtained from the control and treated broilers. Values of P ˂ 0.05 were considered significant.

Data are presented as means ± SD. Two-way ANOVA (P < 0.05) with Bonferroni post hoc test was used to analyze data. Statistics was performed with GraphPad Prism 4.0 for Windows (GraphPad Software, San Diego, CA, USA). For Real Time qPCR analysis, significance was tested using the “Pair Wise Fixed Reallocation Randomisation Test”, developed by REST software [43 (link)]. The number of experiments is reported in figure legends.

Statistical analyses of viability experiments were conducted using GraphPad Prism 4.0 for Windows (GraphPad Software, San Diego, CA), using one-way ANOVA with Bonferroni’s multiple comparison post-test. Results were considered significant at p ≤ 0.05.

Data were analyzed for normality and homogeneity of variance with the Shapiro-Wilk test and Levene’s test, respectively. For the reproductive state (GSI and fertilization success) and offspring morphology, a two-way ANOVA with Bonferroni post hoc test was performed to determine how the biological responses were affected by sampling time and site. Differences in immunopositive bands obtained from densitometric analysis were tested through a one-way ANOVA with Dunnett’s post hoc test. A non-parametric Kruskal-Wallis test (K/S) with Mann-Whitney pairwise as post hoc test was used for the variable “position” that did not satisfy normality assumption for nitrite levels and spawning data. For all tests, significance level was set for p < 0.05. Statistical analysis were performed with Past software ver. 3.11 (http://folk.uio.no/ohammer/past/terms.html). Graphics were created with GraphPad Prism 4.0 for Windows (Graphpad Software, San Diego, CA, USA). For Real Time qPCR analysis, significance was tested using the “Pair Wise Fixed Reallocation Randomisation Test”, developed by REST61 (link).

The unpaired t test was used to analyze the normally distributed, numerical data of two groups. The paired t test was used for statistical analysis between methylation rates before and after density-gradients of each subject. KS normality test and F test was used to test if the data were normally distributed and had equal variance. All analyses were performed using GraphPad Prism 4.0 for Windows (GraphPad Software, CA, USA) and a two-sided P value of 0.05 was considered statistically significant.

Data are presented as means ± SD. All normally distributed data were analyzed using one-way ANOVA with Newman-Keuls multiple comparison post-tests when main effects were identified. GAA concentrations were not normally distributed as determined by the D’Agostino Pearson normality test, so non-parametric Kruskal-Wallis analysis was used to determine differences among groups with Dunn’s post-tests when main effects were identified (GraphPad Prism 4.0 for Windows, GraphPad Software, San Diego, CA). Differences were considered significant at P < 0.05.