Metal

Biofilm

Bacteria

Biology

Chromatography

Biochemistry

Polysaccharide

Matrix (chemical analysis)

Genetics

Organic chemistry

Microplastics

Extracellular polymeric substance

Environmental chemistry

Metal toxicity

Heavy metals

Periphyton

Chemistry

Nutrient

Cadmium

Environmental Research

River biofilms inevitably serve as recipients of heavy metals including copper (Cu) and cadmium (Cd) following their introduction in fluvial systems. Nevertheless, the effects of cultivation substrata on the characteristics of river biofilm extracellular polymeric substances (EPS) and the binding behaviors of heavy metals on biofilms remain unclear. Integrating spectroscopic methods with chemometric analyses, we explored the binding behaviors of Cu(II) and Cd(II) onto biofilm EPS cultivated from two representative substrata at the molecular level. Chemical analysis revealed that biofilm cultivated on polyethylene (PE) pieces contained more non-fluorescent protein fractions, whereas EPS from periphyton grown on mineral, i.e., cobblestones was richer in aromatic fractions and polysaccharides. Excitation-emmision matrix combined with parallel factor analysis suggested a stronger interaction between fluorophores in periphytic EPS with Cu(II) compared to fluorophores in plastic biofilm EPS. Integrated use of infrared spectroscopy and two-dimensional correlation analyses revealed that, during the heavy metal binding processes, the amines and phenolics in plastic biofilm EPS gave the fastest responses to metal binding. While the amides and the aliphatic fractions in periphytic EPS showed a preferential binding to heavy metals. This study differentiates the effects of cultivation substrata on structuring the biofilm EPS characteristics and offers new insights into the environmental behaviors of heavy metal discharge into fluvial systems in river biofilm matrix.

Cultivation substrata differentiate the properties of river biofilm EPS and their binding of heavy metals: A spectroscopic insight