Physiological responses of three mono-species phototrophic biofilms exposed to copper and zinc.

In freshwater ecosystem, phototrophic biofilms play a crucial role through adsorption and sequestration of organic and inorganic pollutants. However, extracellular polymeric substance (EPS) secretion by phototrophic biofilms exposed to metals is poorly documented. This work evaluated the physiological responses of phototrophic biofilms by exposing three microorganisms (cyanobacterium Phormidium autumnale, diatom Nitzschia palea and green alga Uronema confervicolum) to 20 and 200 μg L-1 of Cu or 60 and 600 μg L-1 of Zn, both individually and in combination. Analysis of metal effects on algal biomass and photosynthetic efficiency showed that metals were toxic at higher concentrations for these two parameters together and that all the strains were more sensitive to Cu than to Zn. U. confervicolum was the most impacted in terms of growth, while P. autumnale was the most impacted in terms of photosynthetic efficiency. In consequence to metal exposure at higher concentrations (Cu200, Zn600 and Cu200Zn600), a higher EPS production was measured in diatom and cyanobacterium biofilms, essentially caused by an overproduction of protein-like polymers. On the other hand, the amount of secreted polysaccharides decreased during metal exposure of the diatom and green alga biofilms. Size exclusion chromatography revealed specific EPS molecular fingerprints in P. autumnale and N. palea biofilms that have secreted different protein-like polymers during their development in the presence of Zn600. These proteins were not detected in the presence of Cu200 despite an increase of proteins in the EPS extracts compared to the control. These results highlight interesting divergent responses between the three mono-species biofilms and suggest that increasing protein production in EPS biofilms may be a fingerprint of natural biofilm against metal pollutants in freshwater rivers.