Orally administered nano-polystyrene caused vitellogenin alteration and oxidative stress in the Red Swamp Crayfish (Procambarus clarkii)

Nanoplastics (≤100 nm) represent the smallest fraction of plastic litter and may result in the aquatic environment as degradation products of larger plastic material. To date, few studies focused on the interactions of micro- and nanoplastics with freshwater Decapoda. The red swamp crayfish (Procambarus clarkii, Girard, 1852) is an invasive species able to tolerate highly perturbed environments. As a benthic opportunistic feeder, this species may be susceptible to plastic ingestion. In this study, adult P. clarkii, at intermolt stage, were exposed to 100 µg of 100 nm carboxylated polystyrene nanoparticles (PS NPs) through diet in a 72h acute toxicity test. An integrated approach was conceived to assess the biological effects of PS NPs, by analyzing both transcriptomic and physiological responses. Total hemocyte counts, basal and total phenoloxidase activities, glycemia and total protein concentration were investigated in crayfish hemolymph at 0h, 24h, 48h and 72h from PS NPs administration to evaluate general stress response over time. Differentially expressed genes (DEGs) in the hemocytes and hepatopancreas were analyzed to ascertain the response of crayfish to PS NP challenge after 72h. At a physiological level, crayfish were able to compensate for the induced stress, not exceeding generic stress thresholds. The RNA-Sequencing analysis revealed the altered expression of few genes involved in immune response, oxidative stress, gene transcription and translation, protein degradation, lipid metabolism, oxygen demand, and reproduction after PS NPs exposure. This study suggests that a low concentration of PS NPs may induce mild stress in crayfish, and sheds light on molecular pathways possibly involved in nanoplastic toxicity.