Quantifying microplastic ingestion, degradation and excretion in insects using fluorescent plastics

Plastic pollution is a growing threat to our natural environment. Plastic waste/pollution results from high emissions of both macro (>5 mm) and microplastics (MPs; <5 mm) as well as environmental fractioning of macroplastics into MPs. MPs have been shown to have a range of negative impacts on biota. Harmonized methods to accurately measure and count MPs from animal samples are limited, but what methods exist are not ideal for a controlled laboratory environment where plastic ingestion, degradation and elimination can be quantified and related to molecular, physiological and organismal traits. Here, we propose a complete method for isolating and quantifying fluorescent MPs by combining several previously reported approaches into one comprehensive workflow. We combine tissue dissection, organic material digestion, sample filtering and automated imaging techniques to show how fluorescently labelled MPs provided to insects (e.g. in their diet) in a laboratory setting can be isolated, identified and quantified. As a proof of concept, we fed crickets (<i>Gryllodes sigillatus</i>) a diet of 2.5% (w/w) fluorescently labelled plastics and isolated and quantified plastic particles within the gut and frass.