Evidence for multiple mechanisms of toxicity in larval rainbow trout (Oncorhynchus mykiss) co-treated with retene and alpha-naphthoflavone.
Alkylated polycyclic aromatic hydrocarbons, such as retene (7-isopropyl-1-methylphenanthrene), induce cytochrome P450 1A (CYP1A) enzymes and produce dioxin-like toxicity in the embryo-larval stages of fish characterized by the signs of blue sac disease (BSD). The signs of toxicity are well characterized; however, the mechanism is not well understood. To elucidate the role of CYP1A in retene toxicity, larval rainbow trout (Oncorhynchus mykiss) were co-treated with a range of concentrations of alpha-naphthoflavone (ANF), a known CYP1A inhibitor. The co-treatment produced synergistic toxicity at 3.2-100 microg/L ANF, after which toxicity at 180 microg/L ANF dropped to levels typical of retene-only. At 320 microg/L ANF, toxicity increased with or without retene, indicating that ANF alone was capable of inducing BSD. In addition, the additive toxicity of retene-only and 320 microg/L ANF-only approximately equalled that of the co-exposed larvae (100 microg/L retene+320 microg/L ANF), indicating response addition. Thus, two mechanisms of action occurred in co-exposed larvae at different concentrations of ANF. In trout larvae, there was a correlation between toxicity and CYP1A protein concentrations, and in juvenile trout, ANF produced a concentration-dependent inhibition of ethoxyresorufin-O-deethylase (EROD) activity without a measurable drop in CYP1A protein. Taken together, the mechanism underlying the synergistic toxicity is EROD-independent and may be AhR-dependent. This study demonstrated that multiple, exposure-dependent mechanisms can occur in mixture toxicity, suggesting that current risk assessment models may drastically underestimate toxicity, particularly of mixtures containing both CYP1A inducers and inhibitors.
Scott
Jason A
JA
Department of Biology, Queen's University, Kingston, ON, Canada.
Hodson
Peter V
PV
D000818:Animals; D001571:Benzoflavones / Q000633:toxicity*; D015153:Blotting, Western; D019363:Cytochrome P-450 CYP1A1 / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism; D005393:Fish Diseases / Q000139:chemically induced* / Q000201:enzymology; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D017686:Oncorhynchus mykiss; D010616:Phenanthrenes / Q000633:toxicity*; D018336:Receptors, Aryl Hydrocarbon / Q000378:metabolism; D018709:Statistics, Nonparametric; D016019:Survival Analysis; D018675:Toxicity Tests
D001571:Benzoflavones; D010616:Phenanthrenes; D018336:Receptors, Aryl Hydrocarbon; C447880:retene; C011512:alpha-naphthoflavone; D019363:Cytochrome P-450 CYP1A1