Addressing the environmental and health impacts of microplastics requires open collaboration between diverse sectors.

Public concern over the environmental and public health impacts of the emerging contaminant class "microplastics" has recently prompted government agencies to consider mitigation efforts. Microplastics do not easily fit within traditional risk-based regulatory frameworks because their persistence and extreme diversity (of size, shape, and chemical properties associated with sorbed chemicals) result in high levels of uncertainty in hazard and exposure estimates. Due to these serious complexities, addressing microplastics' impacts requires open collaboration between scientists, regulators, and policymakers. Here we describe ongoing international mitigation efforts, with California as a case study, and draw lessons from a similarly diverse and environmentally persistent class of emerging contaminants (per- and polyfluoroalkyl substances) that is already disrupting traditional regulatory paradigms, discuss strategies to address challenges associated with developing health-protective regulations and policies related to microplastics, and suggest ways to maximize impacts of research.

Chlorination of Emerging Contaminants for Application in Potable Wastewater Reuse: Disinfection Byproduct Formation, Estrogen Activity, and Cytotoxicity.

With increasing water scarcity, many utilities are considering the potable reuse of wastewater as a source of drinking water. However, not all chemicals are removed in conventional wastewater treatment, and disinfection byproducts (DBPs) can form from these contaminants when disinfectants are applied during or after reuse treatment, especially if applied upstream of advanced treatment processes to control biofouling. We investigated the chlorination of seven priority emerging contaminants (17ß-estradiol, estrone, 17α-ethinylestradiol, bisphenol A (BPA), diclofenac, p-nonylphenol, and triclosan) in ultrapure water, and we also investigated the impact of chlorination on real samples from different treatment stages of an advanced reuse plant to evaluate the role of chlorination on the associated cytotoxicity and estrogenicity. Many DBPs were tentatively identified via liquid chromatography (LC)- and gas chromatography (GC)-high resolution mass spectrometry, including 28 not previously reported. These encompassed chlorinated, brominated, and oxidized analogs of the parent compounds as well as smaller halogenated molecules. Chlorinated BPA was the least cytotoxic of the DBPs formed but was highly estrogenic, whereas chlorinated hormones were highly cytotoxic. Estrogenicity decreased by ∼4-6 orders of magnitude for 17ß-estradiol and estrone following chlorination but increased 2 orders of magnitude for diclofenac. Estrogenicity of chlorinated BPA and p-nonylphenol were ∼50% of the natural/synthetic hormones. Potential seasonal differences in estrogen activity of unreacted vs reacted advanced wastewater treatment field samples were observed.

Accumulation of HOCs via Precontaminated Microplastics by Earthworm Eisenia fetida in Soil.

Soil is a primary sink for plastics, but the influence of microplastics as carriers on terrestrial cycling of persistent contaminants is poorly understood as compared to aquatic systems. Studies to date have disregarded the potential fact that microplastics are generally contaminated before their entry into soil. In this study, earthworm Eisenia fetida was incubated for 28 d in a soil amended with five common types of microplastics precontaminated with polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) to elucidate contaminant transfer. Accumulation of HOCs in E. fetida varied greatly among different plastic types and HOCs. The freely dissolved concentration (Cfree) of HOCs showed that desorption of HOCs from microplastics into soil was closely related to plastic types and HOC hydrophobicity and was much slower for polystyrene or polypropylene than polyethylene. Biodynamic model analysis suggested that ingestion of microplastics could act as a significant pathway for some microplastics, likely due to HOCs on the plastics being in an "over-equilibrium" state. This was in contrast with mixing clean microplastics into HOC-contaminated soil, where the microplastics decreased bioaccumulation. Therefore, whether microplastics serve as facilitators or inhibitors of HOC bioaccumulation depends on the fugacity gradient of HOCs between microplastics and soil, which highlights the importance of considering the sequence of contamination between the plastics and soil. These findings also question the validity of short-term experiments because of the generally very slow partition kinetics of HOCs on plastics.

Fish and Seabird Gut Conditions Enhance Desorption of Estrogenic Chemicals from Commonly-Ingested Plastic Items.

Plastic is ingested by over 100 bird species and 40 fish species. Once ingested, plastic may release endocrine-disrupting plastic additives in the animal; however, amounts transferred are poorly characterized. We exposed 16 commonly ingested plastic items to fish and seabird laboratory gut mimic models using the digestive enzyme pepsin at pH 2 and shook them for 16 h at either 28 °C (in saltwater) for fish or 40 °C (in freshwater) for seabirds. Gut liquid was then evaluated for estrogen receptor activity using an in vitro cell line, and plastic-additive concentrations were quantified using ultrahigh-performance liquid chromatography/tandem mass spectrometry. Both seabird ( p < 0.0001) and fish gut conditions ( p < 0.0001) significantly enhanced the biological estrogenicity of expanded polystyrene, polyethylene shopping bag, and polypropylene string relative to controls, resulting in up to a 10.6-fold increase in estrogenicity. Out of 12 plastic additives analyzed, bisphenol A (BPA) (204 ± 129%) and diethylhexyl phthalate (DEHP) (175 ± 97%) concentrations were significantly increased in seabird gut conditions relative to control and butylbenzyl phthalate (BBP) (132 ± 68%) was significantly increased in fish gut conditions relative to control. BPA, DEHP, and BBP did not adequately account for the increase in biological estrogenicity, suggesting that uncharacterized plastic additives may have been enhanced by gut conditions.

Impacts of Salinity and Temperature on the Thyroidogenic Effects of the Biocide Diuron in Menidia beryllina.

Diuron is a herbicide used in agricultural and urban settings and also as an antifouling agent. Recent studies have indicated sublethal responses of diuron in the endocrine system of fish and amphibians. Given the potential of climate change to also alter fish endocrinology, the combination of environmental stressors with diuron may contribute to its sublethal toxicity. In this study, the effects of temperature and salinity on thyroid targets of diuron were assessed in juveniles of the estuarine fish Menidia beryllina under different conditions of salinity (10 and 20‰) and temperature (10 and 20 °C). Environmentally relevant concentrations of diuron affected the growth, and the higher temperature reduced the condition factor of animals. Increased levels of T3 were observed in fish from all treatments, and at 10 °C, T4 levels were augmented at 10‰ but reduced at 20‰. Increased gene expression of deiodinases at 20‰ in both temperatures suggests the influence of salinity on the regulation of hormone imbalance via deiodination pathway activation. Decreased transcripts of thyroid and growth hormone receptors were also observed following diuron treatment. These results indicate that changes in environmental stressors may have significant impacts on the ecological risk of diuron in estuarine fish.

Effects of HCO3- on Degradation of Toxic Contaminants of Emerging Concern by UV/NO3.

This study investigated the significant influence of HCO3- on the degradation of contaminants of emerging concern (CECs) during nitrate photolysis at 254 nm for water reuse applications. The second-order rate constants for the reactions between selected contaminants with carbonate radical (CO3•-) were determined at pH 8.8 and T = 20 °C: estrone ((5.3 ± 1.1) × 108 M-1 s-1), bisphenol A ((2.8 ± 0.2) × 108 M-1 s-1), 17α-ethynylestradiol ((1.6 ± 0.3) × 108 M-1 s-1), triclosan ((4.2 ± 1.4) × 107 M-1 s-1), diclofenac ((2.7 ± 0.7) × 107 M-1 s-1), atrazine ((5.7 ± 0.1) × 106 M-1 s-1), carbamazepine ((4.2 ± 0.01) × 106 M-1 s-1), and ibuprofen ((1.2 ± 1.1) × 106 M-1 s-1). Contributions from UV, reactive nitrogen species (RNS), hydroxyl radical (•OH), and CO3•- to the CEC decomposition in UV/NO3- in the presence and absence of HCO3- were investigated. In addition, possible transformation products and degradation pathways of triclosan, diclofenac, bisphenol A, and estrone in UV/NO3-/HCO3- were proposed based on the mass (MS) and MS2 spectra. Significant reduction in the cytotoxicity of bisphenol A was observed after the treatment with UV/NO3-/HCO3-.

A growing plastic smog, now estimated to be over 170 trillion plastic particles afloat in the world's oceans-Urgent solutions required.

As global awareness, science, and policy interventions for plastic escalate, institutions around the world are seeking preventative strategies. Central to this is the need for precise global time series of plastic pollution with which we can assess whether implemented policies are effective, but at present we lack these data. To address this need, we used previously published and new data on floating ocean plastics (n = 11,777 stations) to create a global time-series that estimates the average counts and mass of small plastics in the ocean surface layer from 1979 to 2019. Today's global abundance is estimated at approximately 82-358 trillion plastic particles weighing 1.1-4.9 million tonnes. We observed no clear detectable trend until 1990, a fluctuating but stagnant trend from then until 2005, and a rapid increase until the present. This observed acceleration of plastic densities in the world's oceans, also reported for beaches around the globe, demands urgent international policy interventions.

Training the next generation of plastics pollution researchers: tools, skills and career perspectives in an interdisciplinary and transdisciplinary field.

Plastics pollution research attracts scientists from diverse disciplines. Many Early Career Researchers (ECRs) are drawn to this field to investigate and subsequently mitigate the negative impacts of plastics. Solving the multi-faceted plastic problem will always require breakthroughs across all levels of science disciplinarity, which supports interdisciplinary discoveries and underpins transdisciplinary solutions. In this context, ECRs have the opportunity to work across scientific discipline boundaries and connect with different stakeholders, including industry, policymakers and the public. To fully realize their potential, ECRs need to develop strong communication and project management skills to be able to effectively interface with academic peers and non-academic stakeholders. At the end of their formal education, many ECRs will choose to leave academia and pursue a career in private industry, government, research institutes or non-governmental organizations (NGOs). Here we give perspectives on how ECRs can develop the skills to tackle the challenges and opportunities of this transdisciplinary research field and how these skills can be transferred to different working sectors. We also explore how advisors can support an ECRs' growth through inclusive leadership and coaching. We further consider the roles each party may play in developing ECRs into mature scientists by helping them build a strong foundation, while also critically assessing problems in an interdisciplinary and transdisciplinary context. We hope these concepts can be useful in fostering the development of the next generation of plastics pollution researchers so they can address this global challenge more effectively.

Development and application of a health-based framework for informing regulatory action in relation to exposure of microplastic particles in California drinking water.

Microplastics have been documented in drinking water, but their effects on human health from ingestion, or the concentrations at which those effects begin to manifest, are not established. Here, we report on the outcome of a virtual expert workshop conducted between October 2020 and October 2021 in which a comprehensive review of mammalian hazard studies was conducted. A key objective of this assessment was to evaluate the feasibility and confidence in deriving a human health-based threshold value to inform development of the State of California's monitoring and management strategy for microplastics in drinking water. A tiered approach was adopted to evaluate the quality and reliability of studies identified from a review of the peer-reviewed scientific literature. A total of 41 in vitro and 31 in vivo studies using mammals were identified and subjected to a Tier 1 screening and prioritization exercise, which was based on an evaluation of how each of the studies addressed various quality criteria. Prioritized studies were identified largely based on their application and reporting of dose-response relationships. Given that methods for extrapolating between in vitro and in vivo systems are currently lacking, only oral exposure in vivo studies were identified as fit-for-purpose within the context of this workshop. Twelve mammalian toxicity studies were prioritized and subjected to a Tier 2 qualitative evaluation by external experts. Of the 12 studies, 7 report adverse effects on male and female reproductive systems, while 5 reported effects on various other physiological endpoints. It is notable that the majority of studies (83%) subjected to Tier 2 evaluation report results from exposure to a single polymer type (polystyrene spheres), representing a size range of 0.040 to 20 µm. No single study met all desired quality criteria, but collectively toxicological effects with respect to biomarkers of inflammation and oxidative stress represented a consistent trend. While it was possible to derive a conservative screening level to inform monitoring activities, it was not possible to extrapolate a human-health-based threshold value for microplastics, which is largely due to concerns regarding the relative quality and reliability of current data, but also due to the inability to extrapolate data from studies using monodisperse plastic particles, such as polystyrene spheres to an environmentally relevant exposure of microplastics. Nevertheless, a conservative screening level value was used to estimate a volume of drinking water (1000 L) that could be used to support monitoring activities and improve our overall understanding of exposure in California's drinking water. In order to increase confidence in our ability to derive a human-health-based threshold value in the future, several research recommendations are provided, with an emphasis towards strengthening how toxicity studies should be conducted in the future and an improved understanding of human exposure to microplastics, insights critically important to better inform future risk assessments. Supplementary Information: The online version contains supplementary material available at 10.1186/s43591-022-00030-6.

Alteration of neuro-dopamine and steroid hormone homeostasis in wild Bank voles in relation to tissue concentrations of PFAS at a Nordic skiing area.

Perfluoroalkyl substances (PFAS) are contaminants that are applied in a wide range of consumer products, including ski products. The present study investigated the neuro-dopamine (DA) and cellular steroid hormone homeostasis of wild Bank voles (Myodes glareolus) from a skiing area in Norway (Trondheim), in relation to tissue concentrations of PFAS. We found a positive association between brain DA concentrations and the concentration of several PFAS, while there was a negative association between PFAS and dopamine receptor 1 (dr1) mRNA. The ratio between DA and its metabolites (3,4-dihydroxyphenylacetic acid: DOPAC and homovanillic acid: HVA) showed a negative association between DOPAC/DA and several PFAS, suggesting that PFAS altered the metabolism of DA via monoamine oxidase (Mao). This assumption is supported by an observed negative association between mao mRNA and PFAS. Previous studies have shown that DA homeostasis can indirectly regulate cellular estrogen (E2) and testosterone (T) biosynthesis. We found no association between DA and steroid hormone levels, while there was a negative association between some PFAS and T concentrations, suggesting that PFAS might affect T through other mechanisms. The results from the current study indicate that PFAS may alter neuro-DA and steroid hormone homeostasis in Bank voles, with potential consequences on reproduction and general health.

Steps Scientists Can Take to Inform Aquatic Microplastics Management: A Perspective Informed by the California Experience.

Recent evidence suggests that microplastic particles are pervasive and potentially of great risk to both animal and human health. The California legislature has responded to this information by enacting two new bills that require quantification of microplastics in various media and development of new management strategies to address microplastic pollution. However, there are several scientific gaps that impede the development and implementation of necessary management strategies to address microplastic pollution. In this paper, we use the California experience as a case study to provide perspective on those science gaps, the current barriers to science affecting management, and the actions scientists can take to best ensure their efforts are of greatest value to policymakers and the management community.

Tebuconazole induced oxidative stress related hepatotoxicity in adult and larval zebrafish (Danio rerio).

Tebuconazole is widely used as fungicide and has frequently been detected at elevated concentrations in environmental media. To characterize the potential toxicity of tebuconazole on vertebrate and humans. Using zebrafish as a vertebrate model, the toxic effects in liver that produced by low-toxic concentrations of tebuconazole were assessed in adult zebrafish. We further focused on tebuconazole-induced toxicity and its possible mechanism in larval zebrafish using a hepatotoxicity assay. The induction of oxidative stress in adult fish was evaluated by superoxide dismutase (T-SOD), catalase (CAT), peroxidase (POD), glutathione S-transferase (GST) activity, and the increased aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio. Significantly increased enzyme activity was observed in the liver of male and female fish at both exposure and depuration stage. Exposure to maximum non-lethal (MNLC) concentration of tebuconazole from 72 to 120 h post-fertilization (hpf) affected the liver size and yolk retention in larval zebrafish. Decreased fluorescence intensity was observed in larval Tg(Apo14:GFP) zebrafish, indicating liver degeneration after tebuconazole treated. Histopathological examination confirmed the alterations in liver histoarchitecture in exposed zebrafish. Significant 1.28-fold and 1.65-fold increases in reactive oxygen species levels were observed in juveniles exposed to MNLC and lethal concentration 10 (LC10) group, respectively. The acridine orange staining assay showed that apoptotic cells occurred in the liver regions. These results indicated that tebuconazole exposure resulted in impacts on the ecological risk in fish and vertebrate. Overall, the present study suggested further research in needed to better understand the tebuconazole-induced toxicity mechanism that associated with oxidative stress.

Degradation of contaminants of emerging concern by UV/H2O2 for water reuse: Kinetics, mechanisms, and cytotoxicity analysis.

Advanced oxidation using UV and hydrogen peroxide (UV/H2O2) has been widely applied to degrade contaminants of emerging concern (CECs) in wastewater for water reuse. This study investigated the degradation kinetics of mixed CECs by UV/H2O2 under variable H2O2 doses, including bisphenol A, estrone, diclofenac, ibuprofen, and triclosan. Reverse osmosis (RO) treated water samples from Orange County Water District's Groundwater Replenishment System (GWRS) potable reuse project were collected on different dates and utilized as reaction matrices with spiked additions of chemicals (CECs and H2O2) to assess the application of UV/H2O2. Possible degradation pathways of selected CECs were proposed based on high resolution mass spectrometry identification of transformation products (TPs). Toxicity assessments included cytotoxicity, aryl hydrocarbon receptor-binding activity, and estrogen receptor-binding activity, in order to evaluate potential environmental impacts resulting from CEC degradation by UV/H2O2. Cytotoxicity and estrogenic activity were significantly reduced during the degradation of mixed CECs in Milli-Q water by UV/H2O2 with high UV fluence (3200 mJ cm-2). However, in GWRS RO-treated water samples collected in April 2017, the cytotoxicity and estrogen activity of spiked CEC-mixture after UV/H2O2 treatment were not significantly eliminated; this might be due to the high concentration of target CEC and their TPs, which was possibly affected by the varied quality of the secondary treatment influent at this facility such as sewer-shed and wastewater discharges. This study aimed to provide insight on the impacts of post-UV/H2O2 CECs and TPs on human and ecological health at cellular level.

Negligible effects of microplastics on animal fitness and HOC bioaccumulation in earthworm Eisenia fetida in soil.

As one type of the most widespread and long-lasting anthropogenic contaminants, microplastics have become a global environmental concern. While numerous studies have demonstrated effects of microplastics on aquatic organisms, the potential influence on terrestrial faunas is relatively less known, even though soil is a primary recipient and sink of plastics. In this study, earthworm Eisenia fetida was exposed to different levels (0, 1, 5, 10, and 20% d.w.) of polyethylene (PE, ≤300 µm) and polystyrene (PS, ≤250 µm) particles in an agricultural soil to evaluate the oxidative stress. Fluorescence imaging, after dying with Nile Red, clearly indicated the ingestion of PE and PS particles by E. fetida. Exposure to PE or PS particles at the highest rate (20%) for 14 d significantly (p < 0.05) increased the activity of catalase and peroxidase and the level of lipid peroxidation, while inhibited the activity of superoxide dismutase and glutathione S-transferase in E. fetida. However, no discernible effect was detected at amendment rates ≤10% for the majority of biochemical endpoints, suggesting that microplastic-induced oxidative stress would not occur in E. fetida under most environmental conditions. The influence of microplastics on bioaccumulation of PAHs and PCBs was also evaluated in E. fetida exposed to different levels (0, 0.1, 1, 5, and 10% d.w.) of PE and PS particles. The tissue concentrations of PAHs and PCBs were reduced in the presence of microplastics at amendment rates ≥1%, suggesting that microplastics did not act as a carrier to enhance contaminant uptake. This was attributed to competitive sorption of microplastics for contaminants and the specific feeding behavior of earthworm. Biodynamic model analysis confirmed that ingestion of microplastics contributed negligibly to contaminant bioaccumulation. Findings of this study suggested that under environmentally relevant conditions, microplastics should not cause significant toxic effects to E. fetida, nor enhance its accumulation of hydrophobic contaminants.

Simulated digestion of polystyrene foam enhances desorption of diethylhexyl phthalate (DEHP) and In vitro estrogenic activity in a size-dependent manner.

Marine polychaetes and fish are known to ingest polystyrene microparticles in the environment. Laboratory microplastic feeding experiments have demonstrated that plastic may release endocrine-disrupting compounds such as diethylhexyl phthalate (DEHP), which can cause adverse effects in both vertebrates and invertebrates. In order to determine the influence of size and digestive conditions on the desorption of DEHP and other plasticizers to polychaetes and fish, we exposed polystyrene particles of various sizes under invertebrate and vertebrate digestive conditions (vertebrate mimic; pepsin, pH = 2.0, 24 °C, invertebrate mimic; Na taurocholate pH = 7, 18 °C). Estrogen receptor activation and concentrations of 12 plasticizers were measured in the extracts. DEHP, bisphenol S and 4-tert-octylphenol were the only compounds detected. Simulated vertebrate gut digestion did not significantly enhance the release of chemicals nor estrogenic activity. However, a 6.3 ±â€¯2.0-fold increase in the concentration of DEHP was observed in extracts from invertebrate gut conditions (Mean ±â€¯SD; N = 24, p < 0.0001). Additionally, estimated particle surface area was positively correlated with estrogenic activity across all treatment types (r = 0.85, p < 0.0001). Overall, these data indicate an elevated bioaccessibility of DEHP may occur in invertebrates, and size-dependent desorption of uncharacterized estrogenic compounds from plastic suggest additional complexity when considering the risks of MP to aquatic organisms.

Disruptive effects of two organotin pesticides on the thyroid signaling pathway in Xenopus laevis during metamorphosis.

Organotin compounds are the ubiquitous environmental pollutants due to their wide industrial and agricultural applications and unexpected releasing into the environment, which show characteristic of endocrine disruptors to interfere with the synthesis, receptor binding or action of endogenous-hormones. Organotin pesticides (OTPs) are used in agriculture and may impact endocrine functions on organisms. Thyroid hormones (THs) play fundamental roles in regulating the basal metabolism and energy balance, while thyroid function can be impaired by environmental contaminants. Therefore, it is crucial to clarify the effects and mechanisms of OTPs on hypothalamus-pituitary-thyroid (HPT) axis. In this study, Xenopus laevis tadpoles at stage 51 were exposed to fentin hydroxide and fenbutatin oxide (0.04, 0.20 and 1.00 µg·L-1) for 21 days. It was found that both compounds caused inhibitory effects on metamorphic development of tadpoles (e.g., significant decrease in hindlimb length and retarding development). Triiodothyronine (T3) significantly decreased in tadpoles exposed to 0.20 µg/L and 1.00 µg/L of the two OTPs for 14 days or 21 days. The expressions of TH responsive genes trß, bteb and dio2 were down-regulated, while tshß and slc5a5 were up-regulated. Surface plasmon resonance (SPR) binding assays showed that fentin hydroxide had a moderate affinity to recombinant human thyroid hormone receptor ß but fenbutatin oxide did not have. Result of the SPR assay was highly consistent with the luciferase reporter gene assays that fentin hydroxide suppressed the relative luciferase activity in the presence of T3 while fenbutatin oxide did not, demonstrating fentin hydroxide but not fenbutatin oxide displayed an antagonistic activity against T3-TR complex mediated transcriptional activation. Overall, the findings elucidated the mechanisms induced by OTPs along HPT axis. These results highlighted the adverse influences of organotin pesticides on thyroid hormone- dependent development in vertebrates and the need for more comprehensive investigations of their potential ecological risks.

Parental exposure to tebuconazole causes thyroid endocrine disruption in zebrafish and developmental toxicity in offspring.

Azole fungicides are one class of the most extensively applied current-use pesticides. Tebuconazole is a common azole fungicide that has been frequently detected in aquatic ecosystems, thus raising concerns about its ecological safety. However, adverse effects of tebuconazole remain largely unknown, especially with regard to endocrine function in aquatic organisms. In the present study, sexually immature zebrafish were exposed to different concentrations of tebuconazole (0.05, 0.20 and 0.50 mg/L) for 60 days in order to test for transgenerational toxicity on the thyroid endocrine system. Thyroid hormone homeostasis, neuronal, and cardiovascular development were investigated in the F1 generation, which were reared in tebuconazole-free water. In the F0 generation, exposure to 0.20 and 0.50 mg/L tebuconazole reduced both thyroxine (T4) and 3,5,3'-triiodothyronine (T3) levels in females, while the T3 levels were unchanged in males. Decreased heart rate was found in F1 larvae, as well as diminished T4 levels in F1 eggs/larvae. We also observed significantly increased expression of ugt1ab mRNA in two generations of zebrafish. Moreover, expression of mRNA associated with neuronal development (e.g. α1-tubulin, mbp, gap43) and cardiovascular development (e.g. cacna1ab, tnncal) were significantly downregulated in F1 larvae at 5 and 10 dpf. In addition, tebuconazole was detected in F1 eggs following parental exposure, indicating maternal transfer. This study demonstrated that tebuconazole can be transferred to offspring from exposed parents, causing thyroid endocrine disruption and developmental toxicity.

Comparisons of analytical chemistry and biological activities of extracts from North Pacific gyre plastics with UV-treated and untreated plastics using in vitro and in vivo models.

Plastic debris is an emerging worldwide threat to marine biota. Marine species may face unique challenges in low-flow estuarine systems with a high abundance of "macro-sized" (>4.75 mm) plastic due to the leaching of constituents and adsorbed contaminants. To simulate this leaching process, plastic samples recovered from the North Pacific Gyre along with corresponding UV-irradiated virgin plastic and non-irradiated virgin plastic counterparts were incubated in saltwater for 30 days at ambient temperatures ranging from 17 to 25 °C. Following solid-phase extraction, water samples were fractionated with sequential methanol elution from 10 to 100% and evaluated using in vitro assays assessing estrogen receptor (ER) and aryl hydrocarbon receptor (AhR) activities. In vivo responses (vitellogenin [vtg] and cytochrome p450 1A [cyp1a] mRNA) were measured following 5-day exposures in Japanese medaka (Oryzias latipes) larvae (3 days post hatch). Estrogenic plasticizers, co-planar PCBs and PAHs were quantified in the extracts using targeted GC-MS/MS and UPLC-MS/MS. In vitro estrogenicity showed highest activity in the 70% methanol fraction for all plastic leachate exposures. Whole extract in vitro estradiol equivalent (EEQ) values were 4.34 ±â€¯2.65, 8.79 ±â€¯2.09 and 13.78 ±â€¯3.64 ng/L, for virgin plastic, UV-irradiated virgin plastic and North Pacific Gyre-recovered plastic, respectively (mean ±â€¯SD). Significant vtg induction was observed in medaka larvae exposed to leachate extracts from North Pacific Gyre-recovered plastic and UV-irradiated virgin plastic (9.9-fold, p = 0.039 and 10.1-fold, p = 0.042, respectively). Chemically-determined EEQ values were also localized in the 70% methanol fraction. Whole leachate extract chemical EEQ values were 0.33 ±â€¯0.07, 1.64 ±â€¯0.62 and 11.4 ±â€¯2.13 ng/L, for virgin plastic, UV-irradiated virgin plastic and North Pacific Gyre-recovered plastic, respectively. In-vitro AhR activity was highest in the 70% methanol elution with greater activity in North Pacific Gyre-recovered plastic than in virgin plastic and UV-irradiated virgin plastic (toxic equivalency [TEQ] = 1.06 ±â€¯0.54, 0.38 ±â€¯0.07 and 0.71 ±â€¯0.47 ng/L, respectively). CYP1A mRNA was significantly induced in larval medaka exposed to North Pacific Gyre-recovered plastic leachates (17.8-fold, p = 0.02) while exposure to virgin plastic and UV-irradiated virgin plastic leachates caused no significant change. Chemically-determined TEQ analysis for AhR indicated highest activity in the 90% methanol fraction for all leachates, with whole extract in vitro TEQs being 1.47 ±â€¯0.87, 0.03 ±â€¯0.05 and 0.42 ±â€¯0.38 ng/L for North Pacific Gyre-recovered plastic, virgin plastic and UV-irradiated virgin plastic, respectively. These results indicate that weathering and UV radiation release estrogenic plasticizers and demonstrate the ability for plastics to transport adsorbed persistent organic pollutants at eco-toxicologically relevant concentrations.