Assessment of the effect of silver nanoparticles on the relevant soil protozoan genus Acanthamoeba

Silver nanoparticles (AgNP) are used in a broad range of consumer products and industrial applications. During the regular product life cycle and disposal, AgNP are continuously released into the environment. Hence, the aim of this study was to investigate the potential ecotoxicological effects of AgNP exposure on amoebae. The Acanthamoeba castellanii ATCC 30234 strain and environmental isolate Acanthamoeba strain C5/2, which are both affiliated with genotype T4, were chosen as representatives of ecologically important soil protozoan organisms. The amoebae were exposed to citrate-stabilized AgNP (30 and 70 nm in size) for 24 h and 96 h at concentrations ranging from 600 mu g L-1 to 20 mg L-1. A newly adopted cell culture based microscopic assay was applied to assess the adherence ability of the amoeba trophozoites. The general metabolic activity of Acanthamoeba was determined to be a second independent endpoint by means of intracellular reduction of the redox dye AlamarBlue((R)). The fate of AgNP within the amoebae and test solutions was visualized by light- and transmission electron microscopy (TEM). Both Acanthamoeba strains showed a significant dose-dependent decrease of adherence ability (p<0.04) and metabolic activity (p < 0.01) after 96 h of AgNP exposure. The environmental Acanthamoeba strain C5/2 lost both its adherence ability and metabolic activity at lower AgNP concentrations than the type strain, indicating a higher sensitivity to ionic silver. This was confirmed by the application of AgNO3, provoking a higher effect level in strain C5/2. AgNP was visualized intracellularly by transmission electron microscopy within the cytoplasm of Acanthamoeba. This is the first report to show the ecotoxicological effects of short-term AgNP exposure on the soil protist Acanthamoeba, causing both changes in the adherence ability and metabolic activity of this amoeba. This combined approach may be a powerful tool in the future for predicting potential harmful ecotoxicological effects of AgNP exposure using soil protozoans.