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We first evaluated the results for the NK 603 feeding trials. The observations shown in Table with relative differences versus controls reveal that of 23 significantly different effects that are supposed to be due to this GM maize, 18 are in males (raw means with SEM; Annex Table ). The repartition of effects is thus sex-dependent. In addition, in general liver (Fig. ) and kidney (Fig. ) parameters in all rats are sex differentially expressed. This is evident not only in the experiments involving NK 603, independently of the treatment at week 14, but also at week 5 (data not shown), but similarly observed in the MON 810 and MON 863 feeding tests (Annex Fig. - Fig. ).

Males are clearly more sensitive than female animals to show physiological disturbances when fed NK 603. This is not observed for all three GM maize varieties. Moreover, most effects appear to be dose-dependent since 83% of male effects emerge only at the 33% feeding level (15/18), the highest GM maize concentration in the diet (Table ). The maximal mean differences are observed in male kidney parameters.

Urine phosphorus, for instance, is importantly disturbed in a dose-dependent manner and at both 5 and 14 week periods of feeding and hence reproducible over time. The significant effect at this level does not appear to be a false positive result (week 5, 33%, adjusted p<0.003 for FDR calculated according to Benjamini and Yekutieli), considering that all parameters were not independent. Comparable results were also obtained for relative lymphocyte and neutrophil differences (all for males, week 14, 33%, adjusted p<0.005).

Table 1 

Differences between NK 603-fed rats and controls.

Study of the GMO effects, which are indicated by mean differences (%) for each parameter with the corresponding control group per sex and per dose. The significant differences versus controls (*p < 0.05, **p < 0.01), for all the parameters measured in the subchronic feeding tests, are presented. The parameters were grouped by organs according to the sites of synthesis or classical indicators of dysfunction. They were indicated for all groups only if they showed at least for one sex or one diet a significant and relatively ± 5% difference to the mean. The animals were male (m) or female (f) young adult rats fed during 5 or 14 weeks with the GM maize NK 603 (11 or 33% in the diet) and compared with controls fed with a ''substantially equivalent'' isogenic maize line. The parameters were measured for 10 rats, except for the organ weights (20 rats), obtained only at the end of the experiment. In single-boxed numbers, we indicate the statistical differences between GMO-fed rats and controls, which are not found between the mean of the six reference groups and controls. A difference between reference and control groups could indicate an effect of the diet per se. In double-boxed numbers, among the effects due to the GMO, are indicated the statistical differences between the GMO groups and the mean of the six reference groups (which have not even eaten a genetically linked variety of maize as the control and the GMO treated groups). (p): Differences for the indicated parameters are not significant by a nonparametric test but by a parametric one; all other differences by both. “Lar Uni Cell” means percent of large unnucleated cell count.

Fig 1 

Principal Component Analysis for liver parameters of all rats in the NK 603 feeding trial. The scheme obtained for parameters at week 14 explains 66.65% of the total data variability (inertia) expressed on 2 axes (49.84% for factor 1; 16.81% for factor 2), scale d=2. This demonstrates the clear separation of parameters values according to sex.

Fig 2 

Principal Component Analysis for kidney parameters of all rats in the NK 603 experiment.

The scheme obtained for parameters at week 14 explains 44.78% of the total data variability (inertia) expressed on 2 axes (27.27% for factor 1; 17.51% for factor 2), scale d=2. This demonstrates the clear separation of parameters values according to sex.

Among 18 GM maize-related effects versus controls, 11 show that groups of reference and control animals are similar in these cases (Table , framed values). However 6 GM-linked effects are also significant versus all reference groups (Table , double framed values). At week 5, these relative maximal effects concern a diminution of blood and increase of urine creatinine clearance, and then a diminution of blood urea nitrogen. This is not observed at week 14 (Fig. a,b). Even so, the kidney parameters measured are clearly the most reactive in both sexes; 52% of significant effects are noticed at this level, but kidney parameters represent only 31% of those measured in total. We also observe that ion concentrations are enhanced in urine of male GM fed rats. Besides this, crude and relative liver weights are also affected at the end of the maximal (33%) GM maize feeding level as well as that of the heart which for corresponding parameters to a comparable extent, showed up to an 11% weight increase. Variations in females are far less frequent (5/23), with no clear significant differences except for urine phosphorus (major relative difference versus controls) and blood potassium (versus all groups).