Do microplastics influence the long-term effects of ciprofloxacin on the polychaete Hediste diversicolor? An integrated behavioral and biochemical approach.

Ciprofloxacin (CPX), the most commonly used fluoroquinolone antibiotic, and microplastics (MPs) are two classes of emerging contaminants with severe adverse impacts on aquatic organisms. Previous studies suggest that both CPX and MPs induce deleterious changes in exposed aquatic biota, but the characterization of a chronic and combined ecotoxicological response is not well known, especially in organisms from estuarine ecosystems. Thus, in this study, we investigated the behavioral and biochemical effects of environmentally relevant levels of CPX alone and in combination with polyethylene terephthalate (PET) microplastics over 28 days of exposure, using the polychaete Hediste diversicolor as a model. In addition to behavioral parameters, different biochemical endpoints were also evaluated, namely the levels of metabolic enzymes of phase I (7-ethoxy-resorufin-O-deethylase, EROD), and phase II (glutathione-S-transferase, GSTs), antioxidant defense (catalase, CAT; glutathione peroxidase, GPx; superoxide dismutase, SOD), oxidative damage (lipid peroxidation, by means of levels of thiobarbituric acid reactive substances [TBARS]) and acetylcholinesterase (AChE). Chronic exposure to ciprofloxacin caused a decrease in burrowing time and a significant increase in SOD activity. In animals exposed to the combination of CPX and PET MPs, effects on behavioral traits were also observed, with higher concentrations of MPs leading to a marked delay in the animals' burrowing time. In addition, these animals showed changes in their antioxidant defenses, namely, a significant increase in SOD activity, while GPx activity was severely compromised. For none of the experimental groups, significant alterations were observed in the metabolic enzymes, TBARS or AChE. These findings provide the first insights into the responses of H. diversicolor when exposed to the combination of CPX and PET MPs, highlighting that, although the here studied conditions, there was no evidence of oxidative damage or neurotoxicity, these organisms are not risk-free in co-exposure scenarios, even at low environmental relevant concentrations.