ABSTRACT
Plastics are widely used in industry and households, but improper disposal has caused their accumulation in aquatic systems worldwide. As a result, mechanical and photochemical processes break down these plastics into microplastics or nano plastics, posing a severe threat to marine organisms and humans as they enter the food chain. This study investigates the effect of Polyvinyl chloride (PVC) and Polyvinyl alcohol (PVA) microplastics in zebrafish by using multi-spectral imaging (MSI), Optical Coherence Tomography (OCT), and Biospeckle OCT (bOCT). These techniques allow for long-term studies in the fish without invasive procedures in real-time. Zebrafish were exposed to Nile red labeled PVC and PVA for 21 days with 500mg/L concentration. Image acquisition and analysis were performed every five days till the end of the study. MSI images revealed deposition of microplastics in the gills region of the fish; some diffused deposition was seen throughout the body in the PVA group towards the end of the experiment. The effect of these MPs on the structure of the gills and their exact location was determined by capturing OCT images. bOCT was used to determine the average speckle contrast for all the OCT images to determine the change in biological activity within the gills region. An increase in bioscpeckle contrast was observed for the MPs treated groups compared to the control group. PVC appeared to cause a more considerable rise in activity compared to PVA. The results indicated that the MPs exert stress on the gills and increase activity within the gills, possibly due to the blockage of the gills and disruption of the water filtration process, which could be monitored non-invasively only by using bOCT. Overall, our study demonstrates the usefulness of non-invasive, robust techniques like MSI, bOCT, and biospeckle for long-term zebrafish studies and real-time analyses.
