This study was to examine single or combined in vitro effects of environmental endocrine disruptors on boar sperm characteristics, oxidative stress damage in sperm and development of porcine IVF embryos. Addition of various concentration of NP (10, 20, 30μM), DBP (10, 50, 100 μM) and BPA (1, 5 or 10 μg/ml) on boar sperm characteristics such as percentages of sperm motility, viability, membrane integrity and mitochondrial activity were dose-dependently decreased within 3, 6 or 9 hr incubation period (p<0.05). The overall detrimental effects increased with incubation time increasement. NP, DBP and BPA showed the detrimental effects on sperm membrane and mitochondria of energy production organelles affecting cell viability with the dependancy of dose and incubation time.In combination effects, NP (10 μM) + DBP (10 μM) significantly decreased boar general sperm characteristics for 3 or 6 hr incubation period compared with control (p<0.05). When both of NP and DBP concentrations (NP; 30 μM, DBP; 100 μM) increase, the detrimental effects on sperm characteristics were larger than those of low concentration combination (p<0.05). The inhibitory effects of NP (30 μM) + BPA (10 μg/ml) on sperm characteristics were larger than those of NP (10 μM) + BPA (1 μg/ml) (p<0.05). DBP (100 μM) + BPA (10 μg/ml) decreased sperm characteristics compared with the low concentration combination (DBP 10 μM + BPA 1 μg/ml, p<0.05). This result indicates the detrimental effects of both chemicals on sperm characteristics were dose dependent. Addition of NP (30 μM) + DBP (100 μM), NP (30 μM) + BPA (10 μg/ml), DBP (10 μM) + BPA (1 μg/ml) or DBP (100 μM) + BPA (10 μg/ml) significantly increased lipid peroxidation for 3 or 6 hr incubation period (p<0.05) compared with no addition control.NP (≥ 20 μM) decreased the percentages of IVF embryo development from morulae and blastocyst stages (p<0.05) and its detrimental effects were dose-dependant. BPA 0, 1, 5 or 10 μg/ml decreased significantly and dose-dependently the percentage of morulae plus and blastocysts (p<0.05). Combinations of DBP (100 μM) plus NP (30 μM) and DBP (100 μM) plus BPA (10 μg/ml) did not affect on morulae and blastocyst development, but NP (30 μM) plus BPA (10 μg/ml) has significant detrimental effect on embryo development at these stages (p<0.05). These overall results indicate that the partial detrimental effects on boar sperm characteristics and embryo development by NP, DBP, BPA or the combination of these chemicals might be due to the increasement of lipid peroxidation and free radical formation in the cell and there were no specific interaction effects on boar sperm and embryo degeneration among the combined treatments.
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