Different species of microorganisms colonize the plastic surfaces and form biofilms depending on the aquatic environment. In the current investigation, characteristics of the plastic surface after exposure to three different aquatic environments based on visualization using scanning electron microscopy (SEM) and spectroscopic (diffuse reflectance (DR) and infrared (IR)) techniques were examined in laboratory bioreactors with time. For both materials, there were no differences observed in the ultraviolet (UV) region among the reactors and several peaks were observed with fluctuating intensities and without any trends. For light density polyethylene (LDPE), peaks indicating the presence of biofilm could be observed in the visible region for activated sludge bioreactor, and for polyethylene terephthalate (PET), freshwater algae biofilm was also visible. PET in freshwater bioreactor is the most densely populated sample both under the optical microscope and SEM. Based on the DR spectra, different visible peaks for LDPE and PET were observed but, in both cases, the visible region peaks (~ 450 and 670 nm) correspond to the peaks found in the water samples of the bioreactors. The difference on these surfaces could not be identified with IR but the fluctuations observed in the UV wavelength region were also detectable using indices obtained from the IR spectra such as keto, ester, and vinyl. For instance, the virgin PET sample shows higher values in all the indices than the virgin LDPE sample [(virgin LDPE: ester Index (I) = 0.051, keto I = 0.039, vinyl I = 0.067), (virgin PET: ester I = 3.5, keto I = 19, vinyl I = 0.18)]. This suggests that virgin PET surface is hydrophilic as expected. At the same time, for all the LDPE samples, all the indices demonstrated higher values (especially for R2) than the virgin LDPE. On the other hand, ester and keto indices for PET samples demonstrated lower values than virgin PET. In addition, DRS technique was able to identify the formation of the biofilm both on wet and dry samples. Both DRS and IR can describe changes in the hydrophobicity during the initial formation of biofilm but DRS can better describe the fluctuations of biofilm in the visible spectra region.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11356-023-27163-2.