Microplastic exposure is associated with epigenomic effects in the model organism Pimephales promelas (fathead minnow).

Microplastics have evolutionary and ecological impacts across species, affecting organisms' development, reproduction, and behavior along with contributing to genotoxicity and stress. As plastic pollution is increasing and ubiquitous, gaining a better understanding of organismal responses to microplastics is necessary. Epigenetic processes such as DNA methylation are heritable forms of molecular regulation influenced by environmental conditions. Therefore, determining such epigenetic responses to microplastics will reveal potential chronic consequences of this environmental pollutant. We performed an experiment across two generations of fathead minnows (Pimephales promelas) to elucidate transgenerational epigenetic effects of microplastic exposure. We exposed the first generation of fish to four different treatments of microplastics: two concentrations of each of pre-consumer polyethylene (PE) and PE collected from Lake Ontario. We then raised the first filial generation with no microplastic exposure. We used enzymatic methylation sequencing on adult liver tissue and homogenized larvae to evaluate DNA methylation differences among treatments, sexes, and generations. Our findings show the origin of the plastic had a larger effect in female minnows whereas the effect of concentration was stronger in the males. We also observed transgenerational effects, highlighting a mechanism in which parents can pass on the effects of microplastic exposure to their offspring. Many of the genes found within differentially methylated regions in our analyses are known to interact with estrogenic chemicals associated with plastic and are related to metabolism. This study highlights the persistent and potentially serious impacts of microplastic pollution on gene regulation in freshwater systems.

Informing the Exposure Landscape: The Fate of Microplastics in a Large Pelagic In-Lake Mesocosm Experiment.

Understanding microplastic exposure and effects is critical to understanding risk. Here, we used large, in-lake closed-bottom mesocosms to investigate exposure and effects on pelagic freshwater ecosystems. This article provides details about the experimental design and results on the transport of microplastics and exposure to pelagic organisms. Our experiment included three polymers of microplastics (PE, PS, and PET) ranging in density and size. Nominal concentrations ranged from 0 to 29,240 microplastics per liter on a log scale. Mesocosms enclosed natural microbial, phytoplankton, and zooplankton communities and yellow perch (Perca flavescens). We quantified and characterized microplastics in the water column and in components of the food web (biofilm on the walls, zooplankton, and fish). The microplastics in the water stratified vertically according to size and density. After 10 weeks, about 1% of the microplastics added were in the water column, 0.4% attached to biofilm on the walls, 0.01% within zooplankton, and 0.0001% in fish. Visual observations suggest the remaining >98% were in a surface slick and on the bottom. Our study suggests organisms that feed at the surface and in the benthos are likely most at risk, and demonstrates the value of measuring exposure and transport to inform experimental designs and achieve target concentrations in different matrices within toxicity tests.

Microplastic and other anthropogenic microparticles in Arctic char (Salvelinus alpinus) and their coastal habitat: A first-look at a central Canadian Arctic commercial fishery.

In the recent monitoring guidelines released by the Arctic Monitoring and Assessment Program's Litter and Microplastic Expert Group, Arctic salmonids were recommended as an important species for monitoring plastics in Arctic ecosystems, with an emphasis on aligning microplastic sampling and analysis methods in Arctic fishes. This recommendation was based on the minimal documentation of microplastics in Northern fishes, especially Arctic salmonids. In response, we worked collaboratively with local partners to quantify and characterize microplastics in Arctic char, Salvelinus alpinus, and their habitats in a commercial fishery near Iqaluktuuttiaq (Cambridge Bay), Nunavut. We sampled Arctic char, surface water, and benthic sediments within their summer foraging habitat at Palik (Byron Bay). We found microplastics in 95 % of char with an average of 26 (SD ± 19) particles per individual. On average, surface water samples had 23 (SD ± 12) particles/L and benthic sediment <1 particles/gww. This is the first documentation of plastic pollution in Arctic char and their coastal habitats. Future work should evaluate seasonal, temporal and spatial trends for long-term monitoring of microplastics in Arctic fishes and their habitats.

A Multicompartment Assessment of Microplastic Contamination in Semi-remote Boreal Lakes.

Microplastic contamination is ubiquitous across the globe, even in remote locations. Still, the sources and pathways of microplastics to such locations are largely unknown. To investigate microplastic contamination in a semi-remote location, we measured microplastic concentrations in nine oligotrophic lakes within and around the International Institute for Sustainable Development-Experimental Lakes Area in northwestern Ontario, Canada. Our first objective was to establish ambient concentrations of microplastics in bottom sediments, surface water, and atmospheric deposition in semi-remote boreal lakes. Across all lakes, mean shallow and deep sediment microplastic concentrations, near-surface water microplastic concentrations from in situ filtering, and dry atmospheric microplastic deposition rates were 551 ± 354 particles kg-1, 177 ± 103 particles kg-1, 0.2 ± 0.3 particles L-1, and 0.4 ± 0.2 particles m-2 day-1, respectively. Our second objective was to investigate whether microplastic contamination of these lakes is driven by point sources including local runoff and direct anthropogenic inputs or nonpoint sources such as atmospheric deposition. Lakes were selected based on three levels of anthropogenic activity-low, medium, and high-though activity levels were minimal across all study lakes compared with highly populated areas. Whereas a positive correlation would indicate that point sources were a likely pathway, we observed no relationship between the level of anthropogenic activity and microplastic contamination of surface water. Moreover, the composition of microplastics in surface water and atmospheric deposition were similar, comprising mostly polyester and acrylic fibers. Together, these results suggest that atmospheric deposition may be the main pathway of microplastics to these remote boreal lakes. Environ Toxicol Chem 2024;43:999-1011. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Microplastics may induce food dilution and endocrine disrupting effects in fathead minnows (Pimephales promelas), and decrease offspring quality.

Microplastics are a complex environmental contaminant that have been reported to cause a variety of impacts, although the mechanism of these impacts remains unclear. Many studies have investigated either sub-organismal or apical endpoints, while very few have attempted to integrate and link endpoints seen at multiple levels of organization. Here, we exposed fathead minnows to microplastics for their entire lifecycle, from the egg stage through to reproduction, and raised a subset of the offspring in clean water. We show that both preconsumer and environmentally sourced microplastics impact adult growth, lipid storage, and external colouration, suggesting a potential food dilution effect. Environmentally sourced microplastics, but not preconsumer microplastics, had further endocrine disrupting impacts on the parental generation and their offspring in the low concentration treatments such that egg production began later, eggs were less viable, and the offspring had higher rates of malformation. Low dose effects are a typical dose-response for endocrine disrupting contaminants. These results suggest that microplastic exposure, at concentrations relevant to what is being found in the environment, has potential implications for forage fish populations. Our findings also highlight the importance of using an integrative approach to understanding the mechanisms behind how and why microplastics impact organisms.

A City-Wide Emissions Inventory of Plastic Pollution.

A global agreement on plastic should have quantitative reduction targets for the emissions of plastic pollution and regular measurements to track success. Here, we present a framework for measuring plastic emissions, akin to greenhouse gas emissions, and demonstrate its utility by calculating a baseline measurement for the City of Toronto in Ontario, Canada. We identify relevant sources of plastic pollution in the city, calculate emissions for each source by multiplying activity data by emission factors for each source, and sum the emissions to obtain the total annual emissions of plastic pollution generated. Using Monte Carlo simulations, we estimate that 3,531 to 3,852 tonnes (T) of plastic pollution were emitted from Toronto in 2020. Littering is the largest source overall (3,099 T), and artificial turf is the largest source of microplastic (237 T). Quantifying source emissions can inform the most effective mitigation strategies to achieve reduction targets. We recommend this framework be scaled up and replicated in cities, states, provinces, and countries around the world to inform global reduction targets and measure progress toward reducing plastic pollution.

High microplastic and anthropogenic particle contamination in the gastrointestinal tracts of tiger sharks (Galeocerdo cuvier) caught in the western North Atlantic Ocean.

Few studies have documented microplastics (<5 mm) in shark gastrointestinal (GI) tracts. Here, we report microplastic contamination in the tiger shark (Galeocerdo cuvier), an apex predator and generalist feeder, at several different life stages. We examined seven stomachs and one spiral valve from eight individuals captured off the United States Atlantic and Gulf of Mexico coasts (eastern US) and conducted a literature review of publications reporting anthropogenic debris ingestion in elasmobranchs. Specimens were chemically digested in potassium hydroxide (KOH) and density separated using calcium chloride (CaCl2) before quantifying and categorizing suspected anthropogenic particles (>45 µm) by size, morphology, and colour. Anthropogenic particles were found in the stomachs and spiral valve of all sharks. A total of 3151 anthropogenic particles were observed across all stomachs with 1603 anthropogenic particles observed in a single specimen. A subset of suspected anthropogenic particles (14%) were chemically identified using Raman spectroscopy and µ-Fourier Transform Infrared spectroscopy to confirm anthropogenic origin. Overall, ≥95% of particles analyzed via spectroscopy were confirmed anthropogenic, with 45% confirmed as microplastics. Of the microplastics, polypropylene (32%) was the most common polymer. Diverse microparticle morphologies were found, with fragments (57%) and fibers (41%) most frequently observed. The high occurrence and abundance of anthropogenic particle contamination in tiger sharks is likely due to their generalist feeding strategy and high trophic position compared to other marine species. The literature review resulted in 32 studies published through 2022. Several methodologies were employed, and varying amounts of contamination were reported, but none reported contamination as high as detected in our study. Anthropogenic particle ingestion studies should continue in the tiger shark, in addition to other elasmobranch species, to further understand the effects of anthropogenic activities and associated pollution on these predators.

Exposure of U.S. adults to microplastics from commonly-consumed proteins.

We investigated microplastic (MP) contamination in 16 commonly-consumed protein products (seafoods, terrestrial meats, and plant-based proteins) purchased in the United States (U.S.) with different levels of processing (unprocessed, minimally-processed, and highly-processed), brands (1 - 4 per product type, depending on availability) and store types (conventional supermarket and grocer featuring mostly natural/organic products). Mean (±stdev) MP contamination per serving among the products was 74 ± 220 particles (ranging from 2 ± 2 particles in chicken breast to 370 ± 580 in breaded shrimp). Concentrations (MPs/g tissue) differed between processing levels, with highly-processed products containing significantly more MPs than minimally-processed products (p = 0.0049). There were no significant differences among the same product from different brands or store types. Integrating these results with protein consumption data from the American public, we estimate that the mean annual exposure of adults to MPs in these proteins is 11,000 ± 29,000 particles, with a maximum estimated exposure of 3.8 million MPs/year. These findings further inform estimations of human exposure to MPs, particularly from proteins which are important dietary staples in the U.S. Subsequent research should investigate additional drivers of MPs in the human diet, including other understudied food groups sourced from both within and outside the U.S.

Microplastics in marine mammal blubber, melon, & other tissues: Evidence of translocation.

Marine mammals consume large quantities of microplastic particles, likely via trophic transfer (i.e., through prey who have consumed plastic) and direct consumption from seawater or sediment. Microplastics have been found in the stomachs, gastro-intestinal tracts, and feces of cetaceans and pinnipeds. Translocation of ingested microplastics has been documented in other organs of several aquatic species, but has not been examined in marine mammals. Marine mammals have highly specialized lipid-rich tissues which may increase susceptibility to lipophilic microplastics. Here we demonstrate the occurrence of microplastics, ranging in size, mass concentration, and particle count concentration from 24.4 µm - 1387 µm, 0.59 µg/g - 25.20 µg/g, and 0.04 - 0.39 particles/g, respectively, in four tissues (acoustic fat pad, blubber, lung, & melon) from twelve marine mammal species inclusive of mysticetes, odontocetes, and phocids. Twenty-two individuals were examined for microplastics using a combination of Raman spectroscopy and pyrolysis gas chromatography with mass spectrometry. Overall, 68% of individuals had at least one microplastic particle in at least one of the four tissue types, with the most common polymer and shape observed being polyethylene and fibers, respectively. These findings suggest some proportion of ingested microplastics translocate throughout marine mammal bodies posing an exposure risk to both marine mammals and people. For people, exposure could be directly through consumption for those who rely on marine mammals as food and indirectly to peoples globally who consume the same prey resources as marine mammals. Some individuals examined represent samples obtained over two decades ago, suggesting that this process, and thus exposure risk, has occurred for some time.

A multi-taxonomic, trait-based framework for assessing macroplastic vulnerability.

Individual interactions with plastic pollution have been documented in hundreds of marine species. However, the population and community level effects of these interactions remain poorly understood. Trait-based approaches provide a method for assessing the relative vulnerability of populations or communities to plastic pollution when empirical studies and data are limited. We conducted a literature review and identified 22 traits that influence likelihood of exposure, species sensitivity, and population resilience to the physical impacts of macroplastic. The resulting trait-based framework provides a process for assessing the relative vulnerability of marine biota to macroplastic ingestion and entanglement. Our framework can be applied to develop vulnerability indices for marine taxonomic groups that can inform targeted management efforts, identify priorities for long-term monitoring, and identify species for future quantitative risk assessments.

The influence of complex matrices on method performance in extracting and monitoring for microplastics.

Previous studies have evaluated method performance for quantifying and characterizing microplastics in clean water, but little is known about the efficacy of procedures used to extract microplastics from complex matrices. Here we provided 15 laboratories with samples representing four matrices (i.e., drinking water, fish tissue, sediment, and surface water) each spiked with a known number of microplastic particles spanning a variety of polymers, morphologies, colors, and sizes. Percent recovery (i.e., accuracy) in complex matrices was particle size dependent, with ∼60-70% recovery for particles >212 µm, but as little as 2% recovery for particles <20 µm. Extraction from sediment was most problematic, with recoveries reduced by at least one-third relative to drinking water. Though accuracy was low, the extraction procedures had no observed effect on precision or chemical identification using spectroscopy. Extraction procedures greatly increased sample processing times for all matrices with the extraction of sediment, tissue, and surface water taking approximately 16, 9, and 4 times longer than drinking water, respectively. Overall, our findings indicate that increasing accuracy and reducing sample processing times present the greatest opportunities for method improvement rather than particle identification and characterization.

Representative subsampling methods for the chemical identification of microplastic particles in environmental samples.

Chemical identification of microplastics is time-consuming, especially when particles are numerous. To save resources, a subsample of particles is often selected for chemical identification. Because no standard subsampling protocols currently exist, methods vary widely and often lack evidence of representativeness, limiting conclusions and cross-study comparability. In this study, we determine best practices for subsampling >100 µm microparticles for chemical identification based on two research objectives: 1) quantifying the proportion of plastic, anthropogenic and natural particles and 2) quantifying the diversity of material types. Using published datasets where all microparticles counted were chemically identified, we tested subsampling methods where particles are selected either from individual samples, or from a group of samples treated collectively. We determine that overall, particle selection at random provides a representative subsample with the lowest effort. Subsampling methods must also be informed by your research objective. Fewer particles are required to accurately represent the proportion of plastic, anthropogenic and natural particles present, compared to representing the diversity of material types. To accurately represent particle diversity, researchers must understand particle diversity within the environmental matrix in question which informs necessary sampling volume. Overall, harmonized, and representative subsampling practices will allow improved comparability among studies, transparent data reporting, and more robust conclusions.

Quantitative assessment of visual microscopy as a tool for microplastic research: Recommendations for improving methods and reporting.

Microscopy is often the first step in microplastic analysis and is generally followed by spectroscopy to confirm material type. The value of microscopy lies in its ability to provide count, size, color, and morphological information to inform toxicity and source apportionment. To assess the accuracy and precision of microscopy, we conducted a method evaluation study. Twenty-two laboratories from six countries were provided three blind spiked clean water samples and asked to follow a standard operating procedure. The samples contained a known number of microplastics with different morphologies (fiber, fragment, sphere), colors (clear, white, green, blue, red, and orange), polymer types (PE, PS, PVC, and PET), and sizes (ranging from roughly 3-2000 µm), and natural materials (natural hair, fibers, and shells; 100-7000 µm) that could be mistaken for microplastics (i.e., false positives). Particle recovery was poor for the smallest size fraction (3-20 µm). Average recovery (±StDev) for all reported particles >50 µm was 94.5 ± 56.3%. After quality checks, recovery for >50 µm spiked particles was 51.3 ± 21.7%. Recovery varied based on morphology and color, with poorest recovery for fibers and the largest deviations for clear and white particles. Experience mattered; less experienced laboratories tended to report higher concentration and had a higher variance among replicates. Participants identified opportunity for increased accuracy and precision through training, improved color and morphology keys, and method alterations relevant to size fractionation. The resulting data informs future work, constraining and highlighting the value of microscopy for microplastics.

Experimental Evidence from the Field that Naturally Weathered Microplastics Accumulate Cyanobacterial Toxins in Eutrophic Lakes.

Freshwater ecosystems with recurring harmful algal blooms can also be polluted with plastics. Thus the two environmental problems may interact. To test whether microplastics influence the partitioning of microcystins in freshwater lakes, we examined the sorption of four microcystin congeners to different polymers of commercially available plastics (low-density polyethylene, polyethylene terephthalate, polyvinyl chloride, and polypropylene). We conducted three experiments: a batch sorption experiment in the laboratory with pristine microplastics of four different polymers, a second batch sorption experiment in the laboratory to compare pristine and naturally weathered microplastics of a single polymer, and a 2-month sorption experiment in the field with three different polymers experiencing natural weathering in a eutrophic lake. This series of experiments led to a surprising result: microcystins sorbed poorly to all polymers tested under laboratory conditions (<0.01% of the initial amount added), irrespective of weathering, yet in the field experiment, all polymers accumulated microcystins under ambient conditions in a eutrophic lake (range: 0-84.1 ng/g). Furthermore, we found that the sorption capacity for microcystins differed among polymers in the laboratory experiment yet were largely the same in the field. We also found that the affinity for plastic varied among microcystin congeners, namely, more polar congeners demonstrated a greater affinity for plastic than less polar congeners. Our study improves our understanding of the role of polymer and congener type in microplastic-microcystin sorption and provides novel evidence from the field, showing that naturally weathered microplastics in freshwater lakes can accumulate microcystins. Consequently, we caution that microplastics may alter the persistence, transport, and bioavailability of microcystins in freshwaters, which could have implications for human and wildlife health. Environ Toxicol Chem 2022;41:3017-3028. © 2022 SETAC.

Local Monitoring Should Inform Local Solutions: Morphological Assemblages of Microplastics Are Similar within a Pathway, But Relative Total Concentrations Vary Regionally.

Pathways for microplastics to aquatic ecosystems include agricultural runoff, urban runoff, and treated or untreated wastewater. To better understand the importance of each pathway as a vector for microplastics into waterbodies and for mitigation, we sampled agricultural runoff, urban stormwater runoff, treated wastewater effluent, and the waterbodies downstream in four regions across North America: the Sacramento Delta, the Mississippi River, Lake Ontario, and Chesapeake Bay. The highest concentrations of microplastics in each pathway varied by region: agricultural runoff in the Sacramento Delta and Mississippi River, urban stormwater runoff in Lake Ontario, and treated wastewater effluent in Chesapeake Bay. Material types were diverse and not unique across pathways. However, a PERMANOVA found significant differences in morphological assemblages among pathways (p < 0.005), suggesting fibers as a signature of agricultural runoff and treated wastewater effluent and rubbery fragments as a signature of stormwater. Moreover, the relationship between watershed characteristics and particle concentrations varied across watersheds (e.g., with agricultural parameters only being important in the Sacramento Delta). Overall, our results suggest that local monitoring is essential to inform effective mitigation strategies and that assessing the assemblages of morphologies should be prioritized in monitoring programs to identify important pathways of contamination.

Monitoring microplastics in drinking water: An interlaboratory study to inform effective methods for quantifying and characterizing microplastics.

California Senate Bill 1422 requires the development of State-approved standardized methods for quantifying and characterizing microplastics in drinking water. Accordingly, we led an interlaboratory microplastic method evaluation study, with 22 participating laboratories from six countries, to evaluate the performance of widely used methods: sample extraction via filtering/sieving, optical microscopy, FTIR spectroscopy, and Raman spectroscopy. Three spiked samples of simulated clean water and a laboratory blank were sent to each laboratory with a prescribed standard operating procedure for particle extraction, quantification, and characterization. The samples contained known amounts of microparticles within four size fractions (1-20 µm, 20-212 µm, 212-500 µm, >500 µm), four polymer types (PE, PS, PVC, and PET), and six colors (clear, white, green, blue, red, and orange). They also included false positives (natural hair, fibers, and shells) that may be mistaken for microplastics. Among laboratories, mean particle recovery using stereomicroscopy was 76% ± 10% (SE). For particles in the three largest size fractions, mean recovery was 92% ± 12% SD. On average, laboratory contamination from blank samples was 91 particles (± 141 SD). FTIR and Raman spectroscopy accurately identified microplastics by polymer type for 95% and 91% of particles analyzed, respectively. Per particle, FTIR spectroscopy required the longest time for analysis (12 min ± 9 SD). Participants demonstrated excellent recovery and chemical identification for particles greater than 50 µm in size, with opportunity for increased accuracy and precision through training and further method refinement. This work has informed methods and QA/QC for microplastics monitoring in drinking water in the State of California.

Conventional and biological treatment for the removal of microplastics from drinking water.

This study examines the removal of microplastics and other anthropogenic particles (>10 µm) from surface water by a full-scale conventional drinking water treatment plant. The treatment process is composed of coagulation with aluminum hydroxide, flocculation, anthracite-sand filtration, and chlorination. Samples were also collected from pilot-scale biological filters consisting of anthracite-sand or granular activated carbon (GAC) media operated with or without pre-ozonation and at a range of different empty-bed contact times (EBCTs). Particles in 10 L water samples collected in duplicate using a fully enclosed sampling apparatus were separated using sieves with 500 µm, 300 µm, 125 µm, and 45 µm openings followed by filtration through 10 µm polycarbonate filters. Particles were counted using stereomicroscopy and characterized using µ-Raman spectroscopy. Full-scale conventional treatment removed 52 % of anthropogenic particles when comparing raw (42 ± 18 particles/L) and finished water (20 ± 8 particles/L). Coagulation, flocculation, and sedimentation accounted for the highest removal (70 %) of any individual unit process. Overall removal was reduced to 52 %, the difference being attributed to airborne particle deposition that occurred while water was detained in a clearwell (exposed to atmosphere via ventilation) that was used to achieve the required contact time for disinfection. The majority of the particles (>80 %) were identified as fibers 10-45 µm; microplastics were predominantly composed of polyester while the non-plastic anthropogenic particles were primarily cellulose. None of the pilot filter configurations examined resulted in significantly fewer microplastics when compared to full-scale conventional filtration. This study illustrates that the removal efficiency of conventional treatment may be limited when considering microfibers <45 µm in size.

Microplastics shift impacts of climate change on a plant-microbe mutualism: Temperature, CO2, and tire wear particles.

Anthropogenic stressors can affect individual species and alter species interactions. Moreover, species interactions or the presence of multiple stressors can modify the stressor effects, yet most work focuses on single stressors and single species. Plant-microbe interactions are a class of species interactions on which ecosystems and agricultural systems depend, yet may be affected by multiple global change stressors. Here, we use duckweed and microbes from its microbiome to model responses of interacting plants and microbes to multiple stressors: climate change and tire wear particles. Climate change is occurring globally, and microplastic tire wear particles from roads now reach many ecosystems. We paired perpendicular gradients of temperature and carbon dioxide (CO2) treatments with factorial manipulation of leachate from tire wear particles and duckweed microbiomes. We found that tire leachate and warmer temperatures enhanced duckweed and microbial growth, but caused effects of microbes on duckweed to become negative. However, induced negative effects of microbes were less than additive with warming and leachate. Without tire leachate, we observed that higher CO2 and temperature induced positive correlations between duckweed and microbial growth, which can strengthen mutualisms. In contrast, with tire leachate, growth correlations were never positive, and shifted negative at lower CO2, again suggesting leachate disrupts this plant-microbiome mutualism. In summary, our results demonstrate that multiple interacting stressors can affect multiple interacting species, and that leachate from tire wear particles could potentially disrupt plant-microbe mutualisms.