An efficient and reproducible method for improving growth of a soil alga (Chlorococcum infusionum) for toxicity assays.

This study evaluated five methods of soil inoculation using the soil alga Chlorococcum infusionum to determine the most efficient and reproducible method for promoting the growth of soil algae for toxicity testing. The five techniques included application of C. infusionum in a circle on top of the soil, to a central spot on top of the soil, to a central spot in the subsoil, to one side on top of the soil, and application divided between a circle and a central spot on top of the soil. Of these, the first method generated the greatest amount of chlorophyll fluorescence and was the method with the best reproducibility. We evaluated the applicability of this method in an assessment of the toxicity of copper and nickel to C. infusionum in two representative standard soils. Copper (20-75 mg/kg for OECD soil and 20-60 mg/kg Lufa 2.2 soil) and nickel (400-500 mg/kg for OECD soil and 60-100 mg/kg Lufa 2.2 soil) reduced the chlorophyll fluorescence of C. infusionum when the inoculation was delivered in a circle on top of both soil types. To our knowledge, this is the first study to assess the suitability of different soil algal inoculation methods for terrestrial toxicity testing.