Assessing applicability of the paper-disc method used in combination with flow cytometry to evaluate algal toxicity

Soil algal bioassays have been limited by their inability to evaluate several toxic endpoints because it is difficult to collect pure soil algae growing on and beneath the soil surface. This study describes the extension of a previously developed paper-disc method for analyzing soil toxicity to algae. The method can be used in conjunction with flow cytometric analysis and facilitates the assessment of previously proposed toxicity endpoints, such as the growth zone, biomass, and photosynthetic activity. We assessed the applicability of this paper-disc soil method using the green algae Chlamydomonas reinhardtii and Pseudokirchneriella subcapitata exposed to nickel-contaminated soil; examined cell sizes, cell granularity, enzyme activity, and oxidative stress as new toxicity endpoints using flow cytometry; and identified morphological changes in green algae assayed. The results showed that, used in conjunction with flow cytometry, the extended paper-disc soil method is sufficiently sensitive to detect decreases in cell granularity in C. reinhardtii and esterase activity in P. subcapitata. The method also revealed decreases in growth zone, biomass, and electron transfer from the reaction center to the quinone pool. Collectively, the results of this study indicate that soil algal bioassays using nonspecific algae can be used to assess soil quality, to derive several toxicity endpoints for individual cells, and to evaluate previously established flow cytometric toxicity endpoints.