Early stage biofilm formation on bio-based microplastics in a freshwater reservoir

Bio-based plastics (BP) produced from renewable biomass resources, such as high-density polyethylene (HDPE), polylactic acid (PLA) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), is currently increasing in terms of both products and applications. However, their biodegradability and environmental fate are not yet fully understood, especially in freshwaters. Here, we present the results of an in-situ study in a freshwater reservoir, where we submerged HDPE, PLA and PHBV microscale BP (mBP) in dialysis bags to enable exchange of small organic and inorganic molecules, including nutrients, with the surrounding water. After one and two months, the bacterial biofilm that formed on each mBP was characterised by 16S rRNA amplicon sequencing. After two-months, Oxalobacteraceae, Pedosphaeraceae, Flavobacteriaceae (Flavobacterium) and Chitinophagaceae (Ferruginibacter) had increased by up to four times. Both these and other common members (≥1 % relative total biomass) of the microbial community were similarly abundant on all mBP. Low-abundance (0.3-1 %) bacterial taxa, however, were significantly more diverse and differed on each mBP. Notably, some low-abundance families and genera increased on specific materials, e.g. Sphingomonadaceae on HDPE, Sphingobacteriaceae on PHBV, Gemmatimonas and Crenothrix on PLA. Overall, abundant bacteria were regarded as a pioneering community, while low-abundance bacteria were more diverse and preferred mBP types in the early stages of biofilm formation on mBP. It could be influenced by the environmental conditions, where nutrient levels and low temperatures might shape the low-abundance of attached bacterial communities than the plastic material itself.