Di-n-butyl phthalate causes estrogenic effects in adult male Murray rainbowfish (Melanotaenia fluviatilis)

•Reduction in the sizes of the spermatocytes and spermatids in fish after treatment with di-n-butyl phthalate (DnBP).•Increase in the proportion of spermatogonia after exposure to DnBP.•Vitellogenin protein induction in 500μg/L and 1000μg/L treatment groups.•An induction in the genes coding for ERα, ERβ, ARα and ARβ after treatment with 1000μ/L DnBP. Phthalic acid esters (PAEs) are a class of synthetic industrial chemicals commonly found in the aquatic environment worldwide. PAEs have been recognised as anti-androgens in male mammals but little is known about their endocrine disrupting effects in fish. This study investigated the effects of 7-day exposures to nominal (measured) concentrations of 125 (62), 250 (140), 500 (230) and 1000 (383)μg/L of di-n-butyl phthalate (DnBP) on the biomarkers of reproduction in adult male Murray River rainbowfish (Melanotaenia fluviatilis) using molecular, biochemical and histological endpoints. None of the tested concentrations of DnBP had any effect on survival or the vital body indices of the fish. The sizes of spermatogonia, Type A and B spermatocytes and spermatids were significantly smaller relative to the controls after treatment with DnBP. This was accompanied by a significant increase in the proportion of spermatogonia in fish treated with 250–1000μg/L of DnBP in comparison to the unexposed fish. At the end of the exposure period, the expressions of the transcripts for the androgen receptors α and β were significantly elevated in the livers of the fish treated with 500 and 1000μg/L of DnBP. In addition, there was also an increase in the circulating concentrations of vitellogenin in the plasma in the higher treatment groups. An induction in the activity of aromatase was noted in the brains of 1000μg/L DnBP-treated fish. This was accompanied by an increase in the hepatic expression of the genes (here and later, whenever the phrase gene expression is used as a synonym for gene transcription although it is acknowledged that it is also regulated, e.g., by translation, mRNA stability and protein stability) encoding for the oestrogen receptors α and β and choriogenin L. Collectively, an increase in the proportion of spermatogonia in the testes, the upregulation of the genes for the oestrogen receptors and choriogenin in the liver, an induction in the brain aromatase activity and the increase in the circulating levels of plasma vitellogenin suggest that continuous exposures for 7 days to sub-acute concentrations of DnBP c