Toxicity assessment of metal mixtures to soil enzymes is influenced by metal dosing method

Abstract Metals are present as mixtures in the environment, yet testing such complex mixture poses design and technical challenges. One possible solution is the use of fixed ratios, i.e. rays of increasing metal concentrations. But fixed ratios rays are compromised when soils dosed with metal salts are leached due to metal-soil selectivity rules. Two alternative metal forms, metal oxides and spinel minerals of quinary metal mixtures (Pb, Cu, Co, Ni, Zn), were evaluated for their toxicity to soil microorganisms measured by the activity of ammonia monooxygenases and acid-phosphatases in three soils. Leaching, a required step for salts, had a larger effect on ammonia monooxygenases than metals. Generally, metal salts were the most toxic form, while the spinel minerals were the least toxic form. Two extractants, CaCl2 and DTPA, were evaluated for their ability to link toxicity to metals across all three metal forms. Salt toxicity was closely linked to CaCl2 extractable concentrations but DTPA was the most appropriate for oxides. We strongly recommend combining fixed ratio rays with metal oxides for metal mixture studies, since soil ratios created using oxides were more precise and required less experimental effort compared to salts and spinel minerals. Furthermore, because DTPA and CaCl2 closely tracked the toxicity of more realistic metal forms (i.e. oxides), we recommend that field studies investigating metal mixtures use both DTPA and CaCl2.