The purpose of this study was to investigate the effect of benzo[a]pyrene (B[a]P) on reproductive endocrine disruption and explore the preliminary mechanisms in crustaceans. In this study, sexually mature female Portunus trituberculatus were exposed to 0, 0.1, 0.5 and 2.5 mu g/L B[a]P for 10 days. The following were investigated: (1) Gonadosomatic Index (GSI) and oocyte diameter, (2) steroid concentrations in ovary and hemolymph, and (3) mRNA levels of genes involved in sex steroid synthesis (3 beta-HSD,17 beta-H5D) or reproduction (estrogen receptor (ER), OUT (Ovarian tumor gene) domain containing ubiquitin aldehyde-binding protein 1 (OTUB1), vitellogenin (VTG),vasa). B[a]P exposure caused significant reductions in the GSI and oocyte diameter in the crabs. Furthermore, 17 beta-estradiol (E-2), testosterone (T) and progesterone (P) levels were inhibited significantly while 3 beta-HSD and 17 beta-HSD mRNA expressions were also decreased in a dose-dependent manner at day 10, which suggests that B[a]P can disrupt sex steroid levels through steroid synthesis pathways. In addition, high levels of B[a]P activated transcription of OTUB1 while suppressed ER and VFG expression, which indicates that exposure to waterborne B[a]P, could interfere with ubiquitin-proteasome pathway and subsequently affect ER and ER-mediated gene expression. We also observed a reduction in vasa gene expression reflecting the negative effect of B[a]P on oocyte development in the molecular level. This study is the first to demonstrate in vivo B[a] toxicity in the reproductive endocrine system of female P. trituberculatus and provided a scientific basis of the decline in crustacean populations. (C) 2015 Elsevier Inc. All rights reserved.