Three generation study of reproductive and developmental toxicity following exposure of pubescent F0 male mice to di-n-butyl phthalate.

Abstract

Humans are exposed to phthalates continuously throughout life. The aim of this study was to evaluate the genotoxic effects induced in male mice following 8 weeks of subchronic exposure to di-n-butyl phthalate (DBP) during their puberty and to investigate the possibility of transmission of mutations to subsequent generations via the sperm. Pzh:Sfis outbred male mice aged 4.5 weeks were exposed to DBP by gavage for 8 weeks, 3 days per week to doses of 1/16 LD50 or 1/4 LD50 each time. Six to seven males from each dosage group were sacrificed at 4, 8 and 12 weeks after the start of exposure for examination of sperm count and quality. Immediately after the end of exposure, the remaining males were caged for 1 week with two unexposed females each. Group of females were sacrificed 1 day before expected parturition, whilst other females were allowed to deliver and rear litters. F1 generation males at 8-9 weeks of age were caged with females from the same group, but from a different litter, for examination of prenatal development of the F2 generation. The remaining F1 generation males were sacrificed at the same age to check the sperm count and quality. Our results confirmed the toxic effects of DBP on the reproductive organs and germ cells of pubertally exposed males. The changes induced in male gametes might be transmitted to the next generation via the sperm. The most important effects were induced in the F1 generation. Exposure of F0 males to DBP induced skeletal malformations in surviving foetuses, caused significant mortality in postnatal life and a disturbance in the sex ratio (superior survival of females in F1), as well as increased frequency of DNA damage in the germ cells of F1 males. The present study did not confirm higher sensitivity to DBP of pubescent males compared to adult males, but the effects induced in the F1 generation differed from that after exposure of adult F0 males.