Effects of single and combined exposure to nano and bulk zinc-oxide and propiconazole on Enchytraeus albidus

Organisms in soil are often exposed to different mixtures of contaminants. These contaminants may interact with each other and, consequently, may have a different effect on organisms than each of them alone. We wanted to investigate possible effects of ZnO mixtures in bulk and nano form and fungicide propiconazole (PCZ) on biochemical parameters and reproduction in Enchytreus albidus. These compounds were applied separately and in binary mixture. In the single exposure experiment the EC₅₀ values for the number of juveniles were calculated: nano ZnO (641.21 ± 88.82 mg kg^-1), bulk ZnO (445.78 ± 148.4 mg kg^-1) and PCZ (3.63 ± 1.68 mg kg-1), respectively. These concentrations were subsequently used in the binary exposure experiment. Calculated combination indices (CI), that allow quantitative determination of chemical interactions at different concentration and effect level, indicated additive or antagonistic interactions (CI ≥ 1) of applied mixture ratios. The only ratio that showed synergistic interaction (CI < 1) was 75% EC₅₀ nZnO/25% EC₅₀ PCZ. Both ZnO forms caused acetylcholinesterase (AChE) activity increase of up to 40% of control level, as well as increased catalase (CAT) and glutathione S-transferase (GST) activities and malondialdehyde (MDA) level. PCZ did not affect AChE and CAT activities, yet it increased GST activity and MDA level. Induced levels of measured biomarkers indicate an oxidative stress after binary exposure, as well. These effects were not enhanced after binary exposure but reflected the effects on biomarkers that corresponding concentrations of these compounds generated in a single exposure experiment.