Evaluation of joint toxicity of heavy metals and herbicide mixtures in soils to earthworms (Eisenia fetida)

It is widely acknowledged that a simplified and robust approach to evaluating thecombined effects of chemical mixtures is critical for ecological risk assessment (ERA) of contaminated soil. The earthworm (Eisenia fetida) was used as a model to study the combined effects of polymetallic contamination and the herbicide siduron in field soil using a microcosm experiment. The responses of multiple biomarkers, including the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and acetylcholine esterase (AChE), the concentrations of glycogen, soluble protein (SP), malonaldehyde (MDA), and metallothionein (MT), and the neutral red uptake test (NRU), were investigated. Multivariate analysis, Principal Component Analysis (PCA) and Spearman's Rank Correlations analysis (BVSTEP) revealed that the activities of AChE and CAT and the NRU content were the prognostic biomarkers capturing the minimum data set of all the variables. Internal Cd (tissue Cd) in earthworms was closely related to the health status of worms under combined contamination of heavy metals and siduron. The integrated effect (E_mix) calculated based on the activities of AChE and CAT and NRU content using the stress index method had significantly linear regression with internal Cd (p<0.01). E_mix(10), E_mix(20), and E_mix(50) were then calculated, at 1.27, 1.63 and 2.71 mg/kg dry weight, respectively. It could be concluded that a bioassay-based approach incorporating multivariate analysis and internal dose was pragmatic and applicable to evaluating combined effects of chemical mixtures in soils under the guidance of the top-down evaluation concept of combined toxicity.