The tests were carried out before and after each experiment, using WEHI-539 hydrochloride following model organisms and standard methods: L. sativum (UNICHIM 1651: 2003 method), P. subcapitata (UNI EN ISO 8692: 2005 method) and V. fischeri (UNI EN ISO 11348-3). To evaluate whether the experimental procedures perturbed the response of the bioassays, the analyses were carried out also on the abiotic control (wastewater without enzymatic inoculum).
Phytotoxicity on L. sativum was expressed as the germination index (GI%), calculated from the following equation:equation(1)GI%=(Gs×Ls)/(Gc×Lc)×100GI%=(Gs×Ls)/(Gc×Lc)×100where Gs is RNA polymerase the mean number of germinated seeds in the sample, Ls is the mean root length of the sample, Gc is the mean number of germinated seeds in the control (i.e. ultrapure water), and Lc is the mean root length of the control.
As regard P. subcapitata assay, the results were expressed as percentage of inhibition of the algal growth (I%) comparing the treated samples with a control made up by nutrient solution only.