Exposure of wild organisms to anthropogenic substances never follows a definite time-course and pulsed events can often determine biological responses to such chemicals, confounding the interpretation of toxicological data. This is the case of specific chemicals such as pharmaceutical drugs, which are commonly released by sewage systems into sensitive areas, including estuaries. The presence and amount of these chemicals in the wild can be modulated by events such as dilution due to heavy rain, floods, or by varying patterns of domestic water use (daily vs. seasonal). The present study aimed to obtain additional data about the toxicity of paracetamol towards the marine clam species Ruditapes philippinarum, following realistic modes of exposure. Thus, the toxicity assessment was made after an acute exposure to different concentrations of paracetamol, followed by a recovery period. The adopted toxicological endpoints included energy-related parameters (glycogen content, GLY; protein content, PROT; electron transport system activity, ETS), activity of antioxidant and biotransformation enzymes (superoxide dismutase, SOD; glutathione peroxidase, GPx; Glutathione-S-transferases, GSTs), levels of reduced glutathione (GSH), neurotoxicity (cholinesterases activity, ChEs), and indicators of oxidative damage (lipid peroxidation, LPO). The here obtained results showed an increase in SOD and GPx activities after exposure. In organisms exposed to the highest concentration tested it was also possible to observe a significant increase in GSTs activity. However, these alterations in the antioxidant defence system were not able to prevent the occurrence of oxidative stress in exposed organisms. Furthermore, exposure to paracetamol induced neurotoxicity in clams, with a concentration-dependent ChEs inhibition along the exposure concentrations. Exposure to paracetamol also led to an increase of GLY content which resulted from metabolic activity depression along the increasing exposure gradient. In recovering organisms the activities of SOD, GPx and GSTs decreased back towards control values presenting lower values than the ones observed in organisms after acute exposure to paracetamol. No LPO was registered in organisms after the recovery period. In addition, after recovery, clams showed no signs of neurotoxicity, with ChEs activities in previously exposed organisms similar to control clams. After recovery clams seemed to re-establish their metabolic capacity, especially evidenced in clams previously exposed to the highest paracetamol concentration as demonstrated by the increase of ETS activity up to control values. Furthermore, the decrease of GLY content after recovery may indicate that clams increased their metabolic activity and started to use their energetic reserves to re-establish their oxidative status. This set of data shows that an acute exposure to paracetamol can exert deleterious effects that may compromise specific biochemical pathways in sensitive aquatic species, such as R. philippinarum, but organisms can re-establish their biochemical status to control levels after a recovery period.
