Zebrafish Vitellogenin ELISA Kit

Complex mixtures of pharmaceutical chemicals in surface waters indicate potential for mixture effects in aquatic organisms. The objective of the present study was to evaluate whether effects on target gene expression and enzymatic activity of individual substances at environmentally relevant concentrations were additive when mixed. Expression of zebrafish cytochrome P4501A (cypla) and vitellogenin (vig) genes as well as activity of ethoxyresorufin-O-deethylase (EROD) were analyzed after exposure (96 h) to caffeine-Caf, ibuprofen-Ibu, and carbamazepine-Cbz (0.05 and 5 mu M), tamoxifen-Tmx (0.003 and 0.3 mu M), and after exposure to pharmaceutical mixtures (low mix: 0.05 mu M of Caf, Ibu, Cbz and 0.003 mu M of Tmx, and high mix: 5 mu M of Caf, Ibu, Cbz and 0.3 mu M of Tmx). Pharmaceuticals tested individually caused significant down regulation of both cypla and vlg, but EROD activity was not affected. Exposure to low mix did not cause a significant change in gene expression; however, the high mix caused significant up-regulation of cypla but did not affect vtg expression. Up-regulation of cypla was consistent with induction of EROD activity in larvae exposed to high mix. The complex mixture induced different responses than those observed by the individual substances. Additive toxicity was not supported, and results indicate the need to evaluate complex mixtures rather than models based on individual effects, since in environment drugs are not found in isolation and the effects of their mixtures is poorly understood.

Portunus trituberculatus broodstock were stocked in plastic tanks to evaluate the effects of starvation and feeding on gonadal development, blood chemistry, fatty acid composition, and expression of vitellogenin (Vtg) and fatty acid-binding protein genes (FABP) in females. Two treatments (starved and fed) were randomly assigned to triplicate groups of 90 swimming crab broodstock (approximately 230 +/- 45 g). In the starved treatment, crabs were starved for 30 days, whereas in the fed treatment crabs were fed once a day with clams. The gonadosomatic index decreased significantly in starved crabs (P < 0.05), as did the serum glucose and cholesterol concentrations; conversely, the total protein concentration in serum significantly increased (P < 0.05). In the ovary, there was a significant relative decline of 18:0, 16:1n-7 and 20:1n-9 fatty acids and relative increases of 20:4n-6, 22:6n3, 18:1n-9 and 20:5n-3 in starved crabs compared to fed crabs (P < 0.05). Relative expression of Vtg in the ovary decreased significantly in starved crabs (P < 0.05), while there was no significant difference in hepatopancreas Vtg expression between starved and fed crabs (P > 0.05). Starvation suppressed gonadal development in female swimming crab broodstock.