Characterization of the transcriptional effects of the plastic additive dibutyl phthalate alone and in combination with microplastic on the green-lipped mussel Perna canaliculus.

The presence and persistence of microplastics (MPs) in diverse aquatic environments are of global concern. Microplastics can impact marine organisms via direct physical interaction and the release of potentially harmful chemical additives incorporated into the plastic. These chemicals are physically bound to the plastic matrix and can leach out. The hazards associated with chemical additives to exposed organisms is not well characterized. We investigated the hazards of plastic additives leaching from plastic. We used the common plasticizer dibutyl phthalate (DBP) as a chemical additive proxy and the New Zealand green-lipped mussel (Perna canaliculus) as a model. We used early-adult P. canaliculus exposed to combinations of virgin and DBP-spiked polyvinyl chloride (PVC), MPs, and DBP alone for 7 days. Whole transcriptome sequencing (RNA-seq) was conducted to assess whether leaching of DBP from MPs poses a hazard. The differences between groups were evaluated using pairwise permutational multivariate analysis of variance (PERMANOVA), and all treatments were significantly different from controls. In addition, a significant difference was seen between DBP and PVC MP treatment. Transcriptome analysis revealed that mussels exposed to DBP alone had the most differentially expressed genes (914), followed by PVC MP + DBP (448), and PVC MP (250). Gene ontology functional analysis revealed that the most enriched pathway types were in cellular metabolism, immune response, and endocrine disruption. Microplastic treatments enriched numerous pathways related to cellular metabolism and immune response. The combined exposure of PVC MP + DBP appears to cause combined effects, suggesting that DBP is bioavailable to the exposed mussels in the PVC MP + DBP treatment. Our results support the hypothesis that chemical additives are potentially an important driver of MP toxicity. Environ Toxicol Chem 2024;43:1604-1614. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Disentangling the influence of microplastics and their chemical additives on a model detritivore system.

Microplastics (MPs) can negatively impact freshwater organisms via physical effects of the polymer itself and/or exposure to chemicals added to plastic during production to achieve desired characteristics. Effects on organisms may result from direct exposure to plastic particles and/or chemical additives or effects may manifest as indirect effects through ecological interactions between organisms (e.g., reduced food availability that impairs a consumer). To disentangle these issues, we used a simplified freshwater food web interaction comprising microbes and macroinvertebrate detritivores to evaluate the toxicity of 1) polyvinyl chloride (PVC) MPs without added chemicals (virgin), 2) the common chemical additive dibutyl phthalate (DBP), and 3) PVC MPs with incorporated DBP. Exposure to virgin PVC MPs (0.33 and 3.3 mg/L) caused negligible ecological effect with the exception of reduced macroinvertebrate feeding rates at 3.3 mg/L. Exposure to DBP (1 mg/L) both individually and when incorporated into the PVC MPs negatively impacted all tested endpoints, including microbial and macroinvertebrate respiration, feeding rate and assimilation efficiency. DBP leached rapidly from the MPs into the water, and also accumulated in macroinvertebrates and their food, providing multiple routes of exposure. Our findings suggest that additives which are intentionally incorporated into MPs could play a key role in MP toxicity and contribute to the disruption of key ecological interactions underpinning ecosystem processes, such as leaf litter decomposition.

Quantifying impacts of titanium dioxide nanoparticles on natural assemblages of riverine phytobenthos and phytoplankton in an outdoor setting.

Impacts of widespread release of engineered titanium dioxide nanoparticles (nTiO2) on freshwater phytoplankton and phytobenthic assemblages in the field, represents a significant knowledge gap. Using outdoor experiments, we quantified impacts of nTiO2 on phytoplankton and periphyton from UK rivers, applied at levels representative of environmentally realistic concentrations (0.05 mg/L) and hot spots of accumulation (5.0 mg/L). Addition of nTiO2 to river water led to rapid temporal size changes in homoagglomerates and many heteroaggregates of nTiO2 with cells in the phytoplankton, including green algae, pennate and centric diatoms, increasing settlement of some cells. Changes in phytoplankton composition were evident after 72-h resulting from a significant decline in the relative abundance of very small phytoplankton cells (1-3 µm), often accompanied by increases in centric diatoms at both concentrations. Significant changes detected in the composition of the phytobenthos after 12 days, following nTiO2 treatments, were not evident when using benthic diatoms alone after 56 days. A lack of inhibition in the maximum quantum yield (Fv/Fm) in phytobenthos after 72-h exposures contrasted with a significant inhibition in Fv/Fm in 75% of phytoplankton samples, the highest recorded in Rutile nTiO2 exposures at both concentrations of nTiO2. After 12 days, strong positive stimulatory responses were recorded in the maximum relative electron transport rate (rETRmax) and the maximum non-photochemical coefficient (NPQmax), in phytoplankton and phytobenthos samples exposed to the higher Anatase nTiO2 concentration, were not measured in Rutile exposed biota. Collectively, these results indicate that the Rutile phase of nTiO2 has more negative impacts on freshwater algae than the Anatase form, at specific time scales, and phytoplankton may be more impacted by nTiO2 than phytobenthos. We caution that repeated release of nTiO2, could lead to significant changes in riverine algal biomass and species composition, dependent on the phase and concentration of nTiO2.

Towards more ecologically relevant investigations of the impacts of microplastic pollution in freshwater ecosystems.

Microplastic pollution is a major environmental concern and the subject of a rapidly growing body of research. Much of this research has focused on the direct effects of microplastics on single species and there is limited information on how microplastics affect different functional groups of organisms, multi-species interactions, and ecosystem processes. We focused on freshwater systems and reviewed 146 studies of microplastic effects on freshwater biota and recorded features including particle characteristics, study designs, functional types of species tested and ecotoxicological endpoints measured. Study species were categorized based on their ecosystem role/functional feeding group rather than taxonomy. We found that most studies were conducted on single species (95%) and focused on a narrow range of functional groups of organisms (mostly filter feeders, 37% of studies). Very few studies have investigated multi-species interactions and ecosystem processes. In many studies, certain characteristics of microplastics, such as polymer type were not well matched with the feeding and habitat ecology of test species, potentially reducing their ecological relevance. Median laboratory study test concentrations were 5-6 orders of magnitude higher than those reported in the field and few studies considered the effects of chemical additives in plastics (6%). We recommend that studies addressing the ecological effects of microplastics need to address neglected functional groups of organisms, design experiments to better match the ecology of test species, and increase in experimental scale and complexity to identify any indirect effects on species interactions and ecosystem processes. We suggest that examining microplastics through an ecological lens that better integrates the feeding and habitat ecology of test organisms will advance our understanding of the effects microplastics have in the environment.