Integrated Analysis of Behavioral and Physiological Effects of Nano-Sized Carboxylated Polystyrene Particles on Daphnia magna Neonates and Adults: A Video Tracking-Based Improvement of Acute Toxicity Assay

Nanoplastics pose significant environmental and public health risks, prompting the need for sensitive, cost-effective, and rapid assays for ecotoxicity assessment. The pre-sent work proposes the use of a portable smartphone-based platform to enhance tradi-tional Daphnia magna acute toxicity assays by integrating behavior analysis and heart rate measurements. The aim is to improve sensitivity in detecting toxic effects of nano-plastics. In particular the study focused on nano-sized carboxylated polystyrene (PS) nanoparticles. Two variability factors that could influence nanoplastic biological effects, the size of the particles and the age of the organisms, were considered. Results demonstrated that the application of the proposed integrated approach allowed the detection of early subtle effects such as a significant impact on the heart rate and be-havior of Daphnia magna under short-term exposure to PS carboxylated nanoparticles. In particular, a stimulation of heart rate was observed for both neonates and adults either for 40 nm or 200 nm particles after 48 h exposure, presumably attributable to an interference of carboxylated PS NPs with adrenergic-type receptors. Behavioral altera-tions were detectable for 40 nm particles but not for 200 nm ones consisting of a de-crease in velocity and alterations of trajectories. Obtained results demonstrated the suitability of the proposed smartphone platform for friendly and real-time integration of behavioral analysis with physiological outcome measurements during acute expo-sure of Daphnia magna to nano-sized carboxylated PS NPs, expanding the sensitivity of the traditional acute toxicity tests. It offers a novel, cost-effective, and field-applicable method for environmental monitoring of nanoparticle toxicity and impact.