Interactive toxicity effects of metronidazole, diclofenac, ibuprofen, and differently functionalized nanoplastics on marine algae Chlorella sp.†

Pharmaceutical products (PPs) and nanoplastics (NPs) are prominent emerging contaminants that pose serious threats to marine ecosystems. The present study aimed to investigate both pristine and combined toxicity of PPs (metronidazole, diclofenac, and ibuprofen) and polystyrene nanoplastics (PSNPs) with amine (NH₂-PSNPs) and carboxyl (COOH-PSNPs) surface functionalization on marine microalgae Chlorella variabilis. Toxicity assessment included the evaluation of growth inhibition, total reactive oxygen species production, malondialdehyde content, antioxidant activity, and photosynthetic activity. Furthermore, changes in the surface functional groups of the algae after exposure to contaminants were examined. The correlation among the toxicity endpoints was assessed using Pearson correlation and cluster heatmap analysis. Zeta potential analysis and hydrodynamic size measurements revealed that the PSNPs became unstable in the presence of PPs. This instability facilitated the aggregation and rapid settlement of PSNPs, consequently impeding their direct interaction with algal cells. Growth inhibition results indicated that Chlorella variabilis exhibited minimal growth inhibition when exposed to pristine PPs (1 mg L⁻¹), whereas PSNPs (1 mg L⁻¹) caused substantial growth inhibition. Notably, the combined toxicity of PSNPs and PPs was lower compared to pristine PSNPs. The independent action model revealed that the combination of PPs and PSNPs showed an antagonistic mode of interaction. The potential reasons for the decreased toxicity observed in the mixture of PSNPs and PPs compared to pristine PSNPs can be attributed to diminished oxidative stress and enhanced photosynthetic activity. These findings provide valuable insights into the role of PPs in modulating the toxicity of PSNPs towards microalgae.