Abstract
This study aimed to assess the ecotoxicological effects of the interaction of fullerene (C60) and benzo[a]pyrene (B[a]P) on the marine mussel, Mytilus galloprovincialis. The uptake of nC60, B[a]P and mixtures of nC60 and B[a]P into tissues was confirmed by Gas Chromatography–Mass Spectrometry (GC–MS), Liquid Chromatography–High Resolution Mass Spectrometry (LC–HRMS) and Inductively Coupled Plasma Mass Spectrometer (ICP–MS). Biomarkers of DNA damage as well as proteomics analysis were applied to unravel the interactive effect of B[a]P and C60. Antagonistic responses were observed at the genotoxic and proteomic level. Differentially expressed proteins (DEPs) were only identified in the B[a]P single exposure and the B[a]P mixture exposure groups containing 1 mg/L of C60, the majority of which were downregulated (~52%). No DEPs were identified at any of the concentrations of nC60 (p < 0.05, 1% FDR). Using DEPs identified at a threshold of (p < 0.05; B[a]P and B[a]P mixture with nC60), gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis indicated that these proteins were enriched with a broad spectrum of biological processes and pathways, including those broadly associated with protein processing, cellular processes and environmental information processing. Among those significantly enriched pathways, the ribosome was consistently the top enriched term irrespective of treatment or concentration and plays an important role as the site of biological protein synthesis and translation. Our results demonstrate the complex multi-modal response to environmental stressors in M. galloprovincialis.
Highlights
There have been concerns regarding the potential for manufactured nanomaterials to cause unpredictable environmental health or hazard impacts, including deleterious effects across differing organismal levels, for over a decade
It is unsurprising that a mussel digestive gland has been used as model tissue for eco-toxicological studies of various NPs [67,68,69], with Di et al reporting that the digestive gland in Mytilus edulis accumulates more C60 than other tissues [67]
Using Danio rerio hepatocytes, Ferreira et al investigated the co-exposure of C60 with B[a]P and provided evidence of toxicological interactions, whereby C60 increased the uptake of B[a]P into cells, decreased cell viability and impaired detoxification responses [75], while Baun et al reported that co-exposure with fullerene C60 enhanced toxicity of phenanthrene to Daphnia magna and Pseudokirchneriella subcapitata [22]
Summary
There have been concerns regarding the potential for manufactured nanomaterials to cause unpredictable environmental health or hazard impacts, including deleterious effects across differing organismal levels, for over a decade. Despite numerous years of study, it is still unclear at what quantity manufactured nanomaterials can be found in the aquatic environment, along with their fate, potential bioavailability and subsequent hazardous effects to biological systems. This is surprising given the growing concern in the field of aquatic toxicology regarding their availability and potential toxicity [1]. Non-functionalised C60 possesses a measurable, but extremely low solubility in water (1.3 × 10−11 mg/mL), but can exist in the aqueous phase as aggregates (nC60) [4] and is quantifiable in aqueous environmental samples [5]. nC60 can be formed in water when fullerenes are released into the aquatic environment, increasing the transport and potential risk of this nanomaterials to the ecosystem ecology
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