Canoco 5 for windows

Data are presented as mean ± SD with N giving the number of animals analyzed. GSH + GSSG concentrations are given as nmol g⁻¹ wwt (wet weight), MDA concentrations as nmol mg⁻¹ protein and enzymes activities as U mg⁻¹ protein. Shapiro–Wilk tests were performed using IBM SPSS Statistics Version 21.0 (IBM Corp., Armonk, NY, USA), to test data of experimental groups for normal distribution. As some groups failed the normality test, non-parametric Mann–Whitney U, Wilcoxon signed rank, and Spearman’s correlation tests were performed in SPSS to detect significant differences between independent groups, between related groups, and correlations, respectively. Redundancy analyses (RDA), which represent principal component analyses (Ter Braak and Šmilauer 2012 ), were performed using the CANOCO 5 for windows software package (Microcomputer Power, Ithaca, NY, USA). Significance of differences was accepted when P < 0.05.

To study the treatment, time and their interaction effects, univariate analysis of variance (ANOVA) was performed through SPSS 17.0 statistical package (IBM, New York, USA). The treatment effects were considered significantly different at 5% probability level. Time dependent effect of nanomaterial on various biochemical parameters was also tested using ANOVA at 5% probability. When main effects among treatments were significant the comparison analysis was performed by using Tukey test. Moreover, relationship of various biochemical parameters among each other and with carbon and nitrogen mineralization were tested for PCA on correlation matrices using Canoco 5 for Windows (Microcomputer Power Inc., Ithaca, NY) after 90 and 180 days of incubation study.