Ecotoxicological effects of atmospheric particulate produced by braking systems on aquatic and edaphic organisms.

Vehicles generate particulate matter (PM) in significant amounts as their brake systems wear. These particles can influence air quality and their transport/deposition may affect the edaphic and aquatic ecosystems. As part of the LOWBRASYS H2020 project, new more eco-friendly brake disc and pad formulations were developed. PMs generated from traditional (FM1-BD1) and innovative (FM4-BD2, FMB-BD7) brake systems in bench tests were studied. The PMs' physical/chemical characteristics were preliminarily investigated. To study the possible environmental impact of the nano-micro particulate, we used a battery of ecotoxicological tests. We employed the microalga Pseudokirchneriella subcapitata, the crustacean Daphnia magna and the bacteria Vibrio fischeri as aquatic bioindicators, while for the edaphic ecosystem we used the seeds of Lepidium sativum and Sorghum saccharatum, the nematode Caenorhabditis elegans, the earthworm Eisenia andrei and the ameba Dictyostelium discoideum. The results showed a higher sensitivity of the freshwater organisms exposed to the soluble PM fraction, with respect to the edaphic ones. FM4-BD2 brake formulation was slightly more toxic for algae (200 mg/L) than FM1-BD1 (500 mg/L). The new system FMB-BD7 particulate was not harmful for crustacean survival, and resulted weakly toxic for algal reproduction only at 500 mg/L. The particulate material per se was found to affect the algal reproduction. No toxic effects were found on nematodes, earthworms and seeds up to 1000 mg/L. However, in D. discoideum the reproduction rate was significantly reduced starting from 100 mg/L; and the lysosomal membrane stability showed a relevant alteration also at minimal concentration (0.1 mg/L). The results demonstrated a minimal risk for biodiversity of the particulates from the different brake systems and highlighted a more eco-friendly performance the new brake-pad FMB-BD7. However, the occurrence of sublethal effects should be considered as a possible contribution of the particle toxicity to the biological effects of the environmental pollution.

Effects of Cr(VI) on Ca2+-ATPase activity in the earthworm Eisenia andrei.

The effect of Cr(VI) as a soil contaminant on the edaphic worm Eisenia andrei was studied by evaluating the activity of Ca2+-ATPase in the intestinal mucosa. In eukaryotes, Ca2+-ATPase is a key mediator of cell signaling although comparatively little is known about its activity in earthworms. Size and anatomical constraints (i.e. small and complex) led us to develop and optimize a cyto-biochemical method to measure Ca2+-ATPase activity in earthworms. The principal site of enzyme activity was found to be the post clitellar intestinal tract; immunohistochemistry then identified plasma membrane Ca2+-ATPase (PMCA ATPase) in the apical area of the intestinal epithelium. Earthworms exposed for 28days to OECD soil contaminated with 1, 2, and 15mg/Kg Cr(VI) demonstrated about 70% inhibition of Ca2+-ATPase activity at the low Cr (VI) concentration (the half of the Italian law limit for residential areas), rising to approximately 84% inhibition at the highest concentration. Reduced enzyme activity was accompanied by decreased enzyme content and reduced lysosomal membrane stability (LMS), which is a well established early warning biomarker of stress. These data demonstrate the potential utility of Ca2+-ATPase activity as a sensitive parameter with which to detect environmental stress in earthworms.