Toxicity, Histopathological Alterations and Immunohistochemical CYP1A Induction in the Early Life Stages of the Seabream, Sparus aurata, Following Waterborne Exposure to B(a)P and TCDD

In this paper, the toxicity (percentage of hatching and LC50) and histopathological alterations induced by benzo(a)pyrene (B(a)P; 0.032, 0.056, 0.1, 0.32, 0.56 and 0.1 microg l(-1)) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 0.025, 0.05, 0.1, 1.5 and 5 pg l(-1)) have been studied in the early life stages of the seabream, Sparus aurata, from 0 to 15 days post-hatching (dph). Toxicity assays showed that the percentage of hatching decreased with increasing the contaminant concentration. Moreover, the number of hatched larvae was lower for TCDD-exposed eggs in comparison with the B(a)P exposed ones. The sensitivity of the larvae, in terms of LC50, towards B(a)P and TCDD increased with age of the larvae. The LC50 were 0.81 microg l(-1) for B(a)P and 4.37 pg l(-1) for TCDD in neonate larvae and 0.11 microg l(-1) for B(a)P and 1.45 pg l(-1) for TCDD in 5 dph larvae. For histopathological examination, samples from LC50 experiments were taken at different concentrations of B(a)P (between 0.032 and 0.1 microg l(-1)) and TCDD (between 0.025 and 5 pg l(-1)). In both, B(a)P- and TCDD-exposed larvae, a concentration-dependency of the histopathological alterations was detected. In contrast, an age-dependency was not clearly detected, possibly due to the lack of development of mostly the organs in the early life stages. Cytoplasmic vacuolization of hepatocytes, as well as subcutaneous edema and necrosis of the trunk musculature, were the most common histopathological disorders detected in both B(a)P- and TCDD-exposed larvae. On the other hand, there were differences in histopathology on exposure to B(a)P and TCDD. Epithelial desquamation in gills, lack of inflation of the swim bladder, as well as lesions in the nervous system were specific for TCDD, while hepatic, vascular and muscular alterations were common for both toxicants. In parallel to the histopathological examinations, immunohistochemical analyses on cytochrome P450-1A isoenzyme (CYP1A) expression were performed on the same samples. The basal/constitutive distribution of CYP1A and its induction was also analysed in similar stages of larval development of the seabream under control conditions and after sublethal exposure to B(a)P (between 0.032 and 0.1 microg l(-1)) and TCDD (between 0.025 and 5 pg l(-1)). During the endogenous nutrition period (from hatching until 4 dph), constitutive CYP1A immunoreactivity was observed in the syncytium and in the matrix of the yolk sac. On the other hand, during exogenous feeding (between 4 and 10 dph), basal CYP1A immunoreactivity was detected in vascular hepatic system, whereas exocrine pancreas showed no reactivity. In gut, basal CYP1A immunoreactivity was restricted to the intestinal brush border and the apical cytoplasm of some enterocytes. Induced CYP1A immunoreactivity in B(a)P-exposed larvae was detected within cytoplasm of hepatocytes, intestinal enterocytes and endothelial cells of the heart. Finally, in TCDD-exposed larvae, CYP1A induction was also detected in pancreatic acinar cells, as well as in renal epithelial cells. The results of this study provided preliminary evidence that constitutive and inducible CYP1A organ distribution in S. aurata larvae was similar to that existing in adult fish. Moreover, exposure to TCDD was more toxic for the larvae and induced more CYP1A that exposure to B(a)P.