The distribution of polycyclic aromatic hydrocarbons (PAHs) in the surface water and sediments in five regions of the Indian Sundarbans was assessed. The capability of microbial biofilm communities to sequester PAHs in a biofilm-promoting vessel was evaluated. The total PAH concentration of water and sediments ranged from undetectable to 125 ng ml⁻¹ and 4880 to 2 × 10⁴ ng g⁻¹ dry weight respectively. The total PAHs concentration of sediments exceeded the Effects Range–Low value and the recommended Effects Range-Median values, implying the PAHs might adversely affect the biota of the Sundarbans. Pyrogenic and petrogenic sources of PAH contamination were identified in most of the sampling sites. Indigenous biofilms were cultivated in a patented biofilm-promoting culture vessel containing liquid media spiked with 16 priority PAHs. Biofilm-mediated 97–100% removal efficiency of 16 PAHs was attained in all media. There was no significant difference between the mean residual PAH from the liquid media collected from hydrophobic and hydrophilic flasks. Residual amounts of acenaphthene (Ace), anthracene (Ant), benzo(b)fluoranthene [B(b)F], benzo(a)pyrene [B(a)P] and benzo(g,h,i)perylene [B(g,h,i)P] showed differences when cultivated in hydrophobic and hydrophilic flasks. The mean residual amounts of total PAHs extracted from biofilm biomasses were variable. A biofilm obtained from a specific sampling site cultured in the hydrophobic flask showed higher PAH sequestration when compared to the removal attained in the hydrophilic flask. Relative abundances of different microbial communities in PAH-sequestering biofilms revealed bacterial phyla including Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria, Chloroflexi and Planctomycetes as well as members of Ascomycota phylum of fungi. The dominance of Candida tropicalis, Clostridium butyricum, Sphingobacterium multivorum and Paecilomyces fulvus were established.