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The aim of this study was to examine ganders at different stages of the reproductive season and the effect of: (1) diets with high phytoestrogen content and (2) in vitro phytoestrogen treatment on testosterone secretion by isolated Leydig cells. Thirty-six male Biłgoraj geese were fed control diets with low phytoestrogen content (containing grass meal) and diets with high phytoestrogens (containing alfalfa meal and soy). Testes were obtained from both groups of ganders at three different times of the breeding season: peak of reproductive activity (March), second half of reproductive activity (May) and beginning of photorefractoriness (July). Isolated Leydig cells were incubated with LH as well as genistein, daidzein, equol and coumestrol and the concentration of testosterone in the medium was determined by radioimmunoassay. The mean weight of testes from ganders in May and July decreased relative to their weights in March, but no significant differences among experimental groups were noted. No differences were observed in basal and LH-stimulated testosterone secretion by Leydig cells of ganders fed the control diets and the diets with higher phytoestrogen content. In July, LH did not stimulate testosterone secretion in either group. In vitro treatment with genistein, daidzein and equol (5 and 50 microM) inhibited basal and LH-stimulated testosterone production by Leydig cells from both groups. Coumestrol (5 and 50 microM) inhibited basal testosterone secretion only in March in the control group. Dietary exposure to phytoestrogens had a slight effect on in vitro testicular secretion in ganders. In vitro treatment with phytoestrogen inhibited testosterone production by Leydig cells. Genistein showed the strongest effect and coumestrol had the weakest influence on testicular secretion.

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The aim of this study was to investigate how long-term feeding of ganders with diets containing different levels of phytoestrogens affects the morphometry of testes, plasma steroid levels, sperm parameters and reproductive performance. Male Bilgoraj geese were fed diets containing grass meal (with low phytoestrogen levels) and those containing alfalfa meal and soybean meal (with higher levels of phytoestrogens). Testes were obtained from ganders at the peak of mating and the ongoing laying period as well as at the beginning of photorefractoriness. The morphometric parameters were measured in frozen testicular sections. Blood was collected monthly from December to June, and plasma testosterone, androstenedione and corticosterone concentrations were determined. During the mating season, semen was collected and its quantity and quality were investigated. In the flocks (ganders with geese) reproductive output was noted. Seasonal changes were observed in the morphology of testes and in plasma testosterone, androstenedione and corticosterone concentrations in ganders. The feeding of ganders with dietary meals containing higher levels of phytoestrogens did not affect the plasma steroid contents or the majority of the morphometric parameters of testes, except for the height of the seminiferous epithelium, which was lowered. Moreover, diets with higher levels of phytoestrogens decreased the volume of ejaculates and the percentage of normal spermatozoa. However, these changes were not reflected in the fertilisation rates.

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The effect of plant-derived, nonsteroidal estrogens (genistein, daidzein and biochanin A) on cortisol and androstenedione secretion by porcine adrenocortical cells was examined. Adrenals were harvested from locally slaughtered mature gilts on days 5-9 of the oestrous cycle. Adrenocortical cells were incubated with or without genistein, daidzein, biochanin A (0.5, 5, 10 or 50 µM), oestradiol (10, 50, 100 or 500 pg/mL) and ACTH (5 nM -positive control). All incubations were performed for 8 h (95% air and 5% CO 2 , 37 °C). ACTH increased cortisol and androstenedione secretion. Genistein, daidzein and biochanin A suppressed cortisol output, whereas androstenedione secretion was enhanced by these phytoestrogens. In contrast, oestradiol did not alter steroid secretion by porcine adrenocortical cells, which may suggest that phytoestrogens have a non-oestrogenic mechanism of action. Therefore, phytoestrogens present in commercial forage may influence adrenocortical function in pigs by decreasing cortisol and increasing androstenedione secretion. Key words: Phytoestrogens, isoflavones, adrenocortical cells, pigPhytoestrogens are phytochemicals present in whole plants, their seeds and plant-derived preserves (Reinli and Block, 1996). These compounds are widely present in legumes and beans, which are the main components of the fodder of farm animals and constitute an integral part of the human diet (Lundh, 1995;Duncan et al., 2003). Soybeans, commonly used in pig diets, are the most significant source of isoflavones (one of the phytoestrogen classes) including

Phytoestrogens (PE) are plant-derived compounds that have an estrogen-like activity and they can influence male and female reproduction. The possible mechanisms of PE action may be including: the binding to estrogen receptors (ER) and the interaction with the key steroidogenic enzymes. The aim of this study was to investigate if PE has effect on steroidogenesis of gander testicular cells by above-described pathways. The Leydig cells were isolated from testes of White Kołuda ganders at the peak of their reproductive activity (March). These Leydig cells (1 × 10(5)per mL) were pre-incubated with the ER inhibitor - ICI 182, 780 (100 nM) for 3 h and then these cells were incubated with PE (5 and 50 μM): genistein, daidzein, equol and coumestrol during next 20 h or untreated control and the Leydig cells that were previously treated (20 h) with genistein (5 and 50 μM) were incubated for next 6 h with steroid intermediates (20 μM) as testosterone (T) precursors: hydroxycholesterol, pregnenolone, progesterone and androstenedione. Concentrations of T in the samples of incubation medium were measured using radioimmunoassay. Genistein, daidzein, and equol (5 and 50 μM) decreased (P < 0.05) T secretion by incubated gander Leydig cells and ICI 182, 780 did not eliminate the inhibitory effect of these PE. After genistein (50 μM) treatment, basal and stimulated with 22R-hydroxycholesterol, pregnenolone, progesterone and androstenedione, T production by testicular cells was decreased (P < 0.05). In contrast, genistein at lower dose (5 μM) did not affect the stimulatory effects of testosterone precursors. In conclusion, the inhibition of testosterone secretion by the phytoestrogens in gander Leydig cells did not depend on estrogen receptors. The suppression of steroidogenesis in these cells may be in part conducted by interaction of phytoestrogens with key steroidogenic enzymes. However, further studies are required to elucidate the phytoestrogen mechanism of action in gander testicular cells.