Similarities and differences among the responses to three chlorinated organophosphate esters in earthworm: Evidences from biomarkers, transcriptomics and metabolomics

The wide use of chlorinated organophosphate esters (Cl-OPEs) as additive flame retardants has aroused concern about their potential risks on ecosystem and human health. However, knowledge about the toxicity of Cl-OPEs on soil organisms remains limited. In this study, earthworms, Eisenia fetida, were exposed to three representative Cl-OPEs, i.e., tris(2-chloroethyl) phosphate (TCEP), tris(2-chloro-1-methylethyl) phosphate (TCPP), and tris(1,3-dichloro-2-propyl) phosphate (TDCPP) in artificial soil. Using a combination of biochemical indicators (biomarkers), transcriptomics, and metabolomics, we compared the Cl-OPE-induced toxicity to E. fetida and provide new insight into the related molecular mechanism. All three Cl-OPEs elicited immune defense by the earthworms, as evidenced by increased acid phosphatase and alkaline phosphatase activities, and the genes involved in immune-related pathways (e.g., lysosomal and interleukin-17 signaling pathways). Furthermore, no effects on acetylcholinesterase activity were observed among the three Cl-OPEs. However, the TCPP and TDCPP treatments significantly decreased the neurotransmitter serotonin, suggesting the potential neurotoxicity of Cl-OPEs. Although TCEP affected the genes involved in carbohydrate and amino acid metabolism, the changes in the corresponding metabolites were not statistically significant. In contrast, exposure to TCPP and TDCPP induced oxidative stress, and affected xenobiotic metabolism and energy metabolism, leading to the decreased body weight in E. fetida. Based on these toxic effects, TCPP and TDCPP were more severely toxic than TCEP, despite their structural similarity. Given that the use of TCEP has been tightly regulated, our results suggest the potentially toxic effects of TCPP and TDCPP should not be ignored in future risk assessments of flame retardants.