| Summary: | Overuse of pesticides in agricultural soil and dye-polluted effluents severely contaminates the environment and is toxic to animals and humans making their removal from the environment essential. The present study aimed to assess the biodegradation of pesticides (cypermethrin (CYP) and imidacloprid (IMI)), and dyes (malachite green (MG) and Congo red (CR)) using biofilms of bacteria isolated from pesticide-contaminated soil and dye effluents. Biofilms of indigenous bacteria, i.e., <i>Bacillus thuringiensis</i> 2A (OP554568), <i>Enterobacter hormaechei</i> 4A (OP723332), <i>Bacillus</i> sp. 5A (OP586601), and <i>Bacillus cereus</i> 6B (OP586602) individually and in mixed culture were tested against CYP and IMI. Biofilms of indigenous bacteria i.e., <i>Lysinibacillus sphaericus</i> AF1 (OP589134), <i>Bacillus</i> sp. CF3 (OP589135) and <i>Bacillus</i> sp. DF4 (OP589136) individually and in mixed culture were tested for their ability to degrade dyes. The biofilm of a mixed culture of <i>B. thuringiensis</i> + <i>Bacillus</i> sp. (P7) showed 46.2% degradation of CYP compared to the biofilm of a mixed culture of <i>B. thuringiensis</i> + <i>E. hormaechei</i> + <i>Bacillus</i> sp. + <i>B. cereus</i> (P11), which showed significantly high degradation (70.0%) of IMI. Regarding dye biodegradation, a mixed culture biofilm of <i>Bacillus</i> sp. + <i>Bacillus</i> sp. (D6) showed 86.76% degradation of MG, which was significantly high compared to a mixed culture biofilm of <i>L. sphaericus</i> + <i>Bacillus</i> sp. (D4) that degraded only 30.78% of CR. UV–VIS spectroscopy revealed major peaks at 224 nm, 263 nm, 581 nm and 436 nm for CYP, IMI, MG and CR, respectively, which completely disappeared after treatment with bacterial biofilms. Fourier transform infrared (FTIR) analysis showed the appearance of new peaks in degraded metabolites and disappearance of a peak in the control spectrum after biofilm treatment. Thin layer chromatography (TLC) analysis also confirmed the degradation of CYP, IMI, MG and CR into several metabolites compared to the control. The present study demonstrates the biodegradation potential of biofilm-forming bacteria isolated from pesticide-polluted soil and dye effluents against pesticides and dyes. This is the first report demonstrating biofilm-mediated bio-degradation of CYP, IMI, MG and CR utilizing soil and effluent bacterial flora from Multan and Sheikhupura, Punjab, Pakistan.
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