Assessment of T-2 effect and its combination with growth factor IGF-I, metabolic hormones leptin and ghrelin on 17β-estradiol secretion by rabbit ovarian fragments was studied. Rabbit ovarian fragments were incubated without (control group) or with alone T-2 toxin (0.01; 0.1; 1; 10; and 100 ng.mL-1), or its combination with insulin-like growth factor I - IGF-I (100 ng.mL-1), leptin (1000 ng.mL-1) or ghrelin (500 ng.mL-1) for 24 hours. Secretion of 17β-estradiol was determined by ELISA. T-2 toxin was not shown to be potential regulator of 17β-estradiol secretion in rabbit ovarian fragments. On the other hand T-2 toxin at all used doses combined with IGF-I significantly (P<0.05) inhibited 17β-estradiol secretion by the fragments. Similarly, 17β-estradiol secretion was significantly (P<0.05) inhibited by T-2 toxin at the highest used doses (10 and 100 ng.mL-1) combined with leptin (1000 ng.mL-1). On the other hand T-2 toxin combined with ghrelin was not shown to be potential regulator of 17β-estradiol secretion in rabbit ovarian fragments. Results in this study showed that trichothecene as T-2 toxin combined with IGF-I or leptin was able to modulate 17β-estradiol secretion in rabbit ovarian fragments in vitro. We suggest their possible involvement to the process of steroidogenesis.