Combined toxicity of triclosan, 2,4-dichlorophenol and 2,4,6-trichlorophenol to zebrafish (Danio rerio)

Triclosan (TCS), 2,4,6-trichlorophenol (2,4,6-TCP) and 2,4-dichlorophenol (2,4-DCP) are the most prevalent chlorinated phenolic pollutants in aquatic environments. Our results showed LC50 and EC50 values of 0.51, 1.11, 2.45 mg/L, and 0.36, 0.74, 1.53 mg/L for TCS, 2,4,6-TCP and 2,4-DCP, respectively, to 120 hpf zebrafish. The highest TCSD (the mixture of TCS, 2,4,6-TCP and 2,4-DCP) toxicity was observed at a TCS:2,4,6-TCP:2,4-DCP concentration ratio of 1:2:4. LC50 and EC50 values of TCSD mixtures for 120-hpf zebrafish were 2.28 and 1.16 mg/L, respectively. Two toxicity assessment methods (Toxic Unit and Mixture Toxicity Index) indicated that TCSD interactions produced partly additive toxicity. TCSD exposure decreased zebrafish hatching rate and led to a series of malformations. Following alkaline phosphatase staining, a large area of vascular ablation was observed with almost complete disappearance of vascular branches and a smaller coverage range. Prominent reddening of the yolk sac and visceral mass after oil red O staining implied that TCSD exposure severely affected fat metabolism. Following acridine orange staining, cell death occurred in eyes while high TCSD concentrations (0.84 mg/L) induced cardiovascular circulation dysfunction. Alcian blue staining increased the alpha angle between Meckel's cartilages and beta angle between two ceratobranchial. Basihyal and palatoquadrate became shorter and developmental abnormality or defects occurred in the fifth ceratobranchial. Overall, these results provide a theoretical basis for systematically evaluating the combined toxicity of the prevalent chlorinated phenolic pollutants in real-world aquatic environments.