Early stage of biofilm formation on bio-based microplastics in a freshwater reservoir
The collected water was passed through 20 m filters to remove other larger unnecessary materials. Then a 500 mL of filtrated water was transferred to each dialysis bag (7.6 x 30 cm) (Merck, Germany). Afterwards, each material was added to the dialysis bag to obtain a final concentration of 1mg/mL. In total 18 dialysis bags were submerged at the same location in the reservoir where the water was taken, each sample was prepared as triplicate. After 1 and 2 months, triplicate samples were withdrawn and filtered through 20 m pore size filters (Whatman cellulose filters, Merck Millipore, Germany) to obtain mBPs with biofilm for DNA extraction and microscopy analysis.