ArticleApplied and environmental microbiology2026
Developing an optimized method for biofilm extraction from microplastic surfaces for high-efficiency analysis of adherent bacterial communities.
Article in Applied and environmental microbiology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Microplastics (MiPs, ×5 mm in size) harbor complex biofilms that facilitate pathogen dissemination, yet standardized extraction protocols are lacking. Here, we developed and optimized a method for biofilm extraction from environmentally weathered MiPs. To reflect real-world conditions, the protocol was applied directly to bulk, heterogeneous, field-collected MiP mixtures (size range: 80 µm-5 mm) without prior sorting by polymer type or morphology. By optimizing extraction buffers, mechanical disruption, and MiP quantities (100-150 particles), we established an optimal protocol combining phosphate-buffered saline with 0.1% Tween 80, ultrasonication (40 kHz, 10 min), vortexing with glass beads, and a two-cycle extraction-disaggregation workflow. This approach involves an initial extraction followed by a repeated, exhaustive extraction step designed to maximize the recovery of recalcitrant biofilm residues. This protocol markedly enhanced recovery of viable, culturable cells, delivering a 2,950-fold enhancement in the recovery of viable, culturable cells (evaluated via CFU counts; 28,020 ± 11,034 CFU MiP⁻¹) vs. conventional PBS extraction (9.5 ± 3 CFU MiP⁻¹) and 102-fold vs. passive extraction (274 ± 59 CFU MiP⁻¹). The 10-min sonication empirically maximized viable cell recovery within the tested duration range. The two-step protocol with Tween 80-mediated disaggregation proved critical, increasing recovery 208-fold by disaggregating biofilm fragments. While DNA yields (26.5 ± 3.93 ng µL⁻¹) were sufficient for targeted PCR-based pathogen detection (
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