Microbenthic biofilms are consortia of autotrophic and heterotrophic organisms imbedded in a matrix of polymers and particles. As biofilms develop, internal cycling of materials might predominate, and dependence on external conditions is reduced. The mature biofilm structure may act as a barrier against deleterious effects of metals on microphytobenthos. To validate this hypothesis, biofilms from two lowland streams near the Dutch-Belgian border, the extremely Zn-and Cd-polluted Eindergatloop and the relatively clean Keersop in the River Dommel subsystem, were collected after 2 weeks (young) and 6 weeks (old) of colonization. Young and old biofilms from both sites were subsequently exposed in the laboratory to Zn and Cd concentrations mimicking that of the heavily polluted stream for a period of 2 weeks. Diatom composition, chlorophyll a, total carbohydrates, Zn and Cd concentrations, minimal chlorophyll fluorescence, and photon yield demonstrated more pronounced metal effects on the young than on the old reference biofilms. In contrast, colonization time had less effect on the overall response of the extremely polluted biofilms. Therefore, biofilms in an early colonization stage are more vulnerable than mature biofilms to metal exposure, and exposure history determines the response of biofilms to metals.
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