Retrieving the real microbial diversity in aquatic plastisphere.

Disposed plastics in oceans provide a substrate to which microbes can adhere and structure the biofilm, namely the plastisphere. In this study, we showed that the mesoplastic density-based separation, routinely used in quantification assays, is detrimental to studying the microbiome diversity and ecology as it underestimates the real microbial diversity within these samples. Based on SEM and microbiome observations, we propose that chemically fixing samples before density separation preserves cellular diversity (2.32-fold change) and richness (1.12-fold change) that would be naturally lost due to the current methodology. OTUs assigned to Gram-negative bacterial species are the most negatively affected by omitting fixation and polymer composition was not decisive in shifting microbiome composition. Considering our findings, the formaldehyde-fixation step should be incorporated into the current methodology described in most studies as this is crucial to promote a deeper understanding of the microbial community in this ecosystem and biofilm-adhered scattering through aquatic ecosystems. • Formaldehyde prefixation before conserves alpha-diversity of mesoplastic community. • Formaldehyde prefixation before conserves beta-diversity of mesoplastic community. • Gram-negative representatives are naturally lost in saline-float method. • Microscopy and metagenomics data corroborate the use of Formaldehyde prefixation. [ABSTRACT FROM AUTHOR]