Harmful algal blooms (HABs), driven by warming oceans and increased eutrophication, are negatively affecting aquatic ecosystems as well as human health. Blooms of diatom Pseudo-nitzschia spp. produce a neurotoxin, domoic acid (DA), which can be concentrated by filter feeders, including shellfish. Consumption of DA-contaminated seafood causes amnesic shellfish poisoning. DA causes well-established effects on neurons, inducing neurotoxicity by binding to ionotropic glutamate receptors. However, its effects on non-neuronal cells including microglia, the resident immune cells in the brain, are less well understood. Microglia play critical roles in brain health, and disruptions in microglial activity during development can have long-term impacts on brain function and disease risk. The objective of this study was to examine the effects of developmental DA exposure on microglia using zebrafish (Danio rerio). We characterized effects of DA exposure on microglial abundance and morphology at two developmental stages (2 and 4 days post-fertilization (dpf)). We additionally assessed impacts on cell death, cytokine expression, and startle response behavior. In older larvae (4 dpf), microglial effects occurred only at the highest dose (0.3 ng DA per larvae) and coincided with severe morphological defects. In contrast, 2 dpf exposure to lower doses (0.1 ng DA per embryo) caused transient tremors without gross abnormalities but exposure to either 0.1 or 0.3 ng DA per larvae increased microglial abundance and altered microglial morphology. In contrast, 4 dpf exposure to 0.3 ng DA per larvae reduced microglial numbers. Increases in mRNA levels of il1b, il4, and tgfb were observed after 4 dpf exposure, but no significant cytokine changes were detected at 2 dpf. Overall, the effects of DA are highly developmental stage-specific, with microglial reactivity occurring at doses that do not cause gross morphological changes. These findings suggest that neuroinflammation may arise at lower DA doses, indicating a potential mechanism for subtle toxic effects and emphasizing the need for further research to better understand the consequences of low-dose DA exposure on microglial function.

Dioxins and dioxin-like compounds (DLCs) exert toxicity through the aryl hydrocarbon receptor (AHR), but species variations in AHR lead to differing sensitivities. Investigating the variation in AHR homolog diversity, expression levels, predominant forms, and AHR sensitivity across species-particularly in fish sensitive to dioxins-is essential for enhancing ecological risk assessment. This study focuses on the tiger puffer (Takifugu rubripes), identifying five AHRs and two ARNTs, with truAHR2a showing the highest expression and the truAHR1 subfamily displaying lower levels. All truAHRs are functional and can be activated by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), with truARNT1 cooperating more efficiently with truAHRs than truARNT2. We determined EC50 values for truAHR1a (0.30 ± 0.10 nM), truAHR1b (0.32 ± 0.20 nM), truAHR2a (0.98 ± 0.63 nM), truAHR2b (2.62 ± 2.48 nM), and truAHR2c (0.43 ± 0.22 nM), with truAHR1a showing the highest sensitivity. The truAHR1 subfamily displayed greater sensitivity than the truAHR2 subfamily, contrasting with medaka and zebrafish, where AHR2 is similar to or more sensitive than AHR1. Comparisons highlighted species- and subform-specific sensitivities in AHRs, differing by one to two orders of magnitude. Ligand-binding assays showed that all truAHRs bound [3H]TCDD specifically. Molecular docking indicated that although TCDD binds AHRs with similar affinities and conserved residues, other subform-specific factors likely contribute to their differential sensitivities. This study provides valuable data on AHR diversity and ligand-sensitivity, contributing to ecological toxicity assessment of dioxin-like compounds.

ABSTRACT The mummichog, Fundulus heteroclitus , an abundant estuarine fish broadly distributed along the eastern coast of North America, has repeatedly evolved tolerance to otherwise lethal levels of aromatic hydrocarbon exposure. This tolerance is linked to reduced activation of the aryl hydrocarbon receptor (AHR) signaling pathway. In other animals, the AHR has been shown to influence the gastrointestinal-associated microbial community, particularly when activated by the model toxic pollutant 3,3′,4,4′,5-pentachlorobiphenyl (PCB-126) and other dioxin-like compounds. To understand host population and PCB-126 exposure effects on mummichog gut microbiota, we sampled two populations of wild fish, one from a PCB-contaminated environment (New Bedford Harbor, MA, USA) and the other from a much less polluted location (Scorton Creek, MA, USA), as well as laboratory-reared F2 generation fish originating from each of these populations. We examined the microbes associated with the gut of these fish using amplicon sequencing of bacterial and archaeal small subunit ribosomal RNA genes. Fish living in the PCB-polluted site had high microbial alpha and beta diversity compared to fish from the low PCB site. These differences between wild fish were not present in laboratory-reared F2 fish that originated from the same populations. Microbial compositional differences existed between wild and lab-reared fish, with the wild fish dominated by Vibrionaceae and the lab-reared fish by Enterococceae. These results suggest that mummichog habitat and/or environmental conditions have a stronger influence on the mummichog gut microbiome compared to population or hereditary-based influences. Mummichog are important eco-evolutionary model organisms; this work reveals their importance for exploring host-environmental-microbiome dynamics. IMPORTANCE The mummichog fish, a common resident of North America's east coast estuaries, has evolved the ability to survive in waters contaminated with toxic chemicals that would typically be deadly. Our study investigates how living in and adapting to these toxic environments may affect their gut microbiomes. We compared mummichogs from a polluted area in Massachusetts with those from a non-polluted site and found significant differences in their gut microbes. Interestingly, when we raised the next generation of these fish in a lab, these differences disappeared, suggesting that the environment plays a more crucial role in shaping the gut microbiome than genetics. Understanding these changes helps shed light on how animals and their associated microbiomes adapt to pollution, which can inform conservation efforts and our broader understanding of environmental impacts on host-microbe dynamics.

Microplastics (MPs) have been found in a diverse range of organisms across trophic levels. While a majority of the information on organismal exposure to plastics in the environment comes from gastrointestinal (GI) data, the prevalence of MP particles in other tissues is not well understood. Additionally, many studies have not been able to detect the smallest, most prevalent, MPs (1 µm – 5 mm) that are the most likely to distribute to tissues in the body. To address these knowledge gaps, MPs in the GI tract and muscle of Atlantic killifish (Fundulus heteroclitus) collected from two sites (Falmouth and Bourne) on Buzzards Bay, Cape Cod, MA were quantified down to 2 µm in size. Eight fish from Falmouth and 10 fish Bourne site were analyzed. Fourier-transform infrared spectroscopy and Raman spectroscopy were used to identify all particles. The mean concentrations of MPs in the GI tract and muscle from fish collected from Falmouth was 85.5 ± 70.2 and 11 ± 12.5 particles per gram wet weight, respectively. Fish collected from Bourne site had mean particle concentrations of 12.2 ± 18.1 and 1.69 ± 5.36 particles per gram wet weight. Of the 2,008 particles analyzed in various fish tissue samples, only 3.4

The genetic architecture of phenotypic traits can affect the mode and tempo of trait evolution. Human-altered environments can impose strong natural selection, where successful evolutionary adaptation requires swift and large phenotypic shifts. In these scenarios, theory predicts that adaptation is due to a few adaptive variants of large effect, but empirical studies that have revealed the genetic architecture of rapidly evolved phenotypes are rare, especially for populations inhabiting polluted environments. Fundulus killifish have repeatedly evolved adaptive resistance to extreme pollution in urban estuaries. Prior studies, including genome scans for signatures of natural selection, have revealed some of the genes and pathways important for evolved pollution resistance, and provide context for the genotype-phenotype association studies reported here. We created multiple quantitative trait locus (QTL) mapping families using progenitors from four different resistant populations, and using RAD-seq genetically mapped variation in sensitivity (developmental perturbations) following embryonic exposure to a model toxicant PCB-126. We found that one to two large-effect QTL loci accounted for resistance to PCB-mediated developmental toxicity. QTLs harbored candidate genes that govern the regulation of aryl hydrocarbon receptor (AHR) signaling. One QTL locus was shared across all populations and another was shared across three populations. One QTL locus showed strong signatures of recent natural selection in the corresponding wild population but another QTL locus did not. Some candidate genes for PCB resistance inferred from genome scans in wild populations were identified as QTL, but some key candidate genes were not. We conclude that rapidly evolved resistance to the developmental defects normally caused by PCB-126 is governed by few genes of large effect. However, other aspects of resistance beyond developmental phenotypes may be governed by additional loci, such that comprehensive resistance to PCB-126, and to the mixtures of chemicals that distinguish urban estuaries more broadly, may be more genetically complex.

Saxitoxin (STX) is a potent neurotoxin naturally produced by dinoflagellates and cyanobacteria. STX inhibits voltage-gated sodium channels (VGSCs), affecting the propagation of action potentials. Consumption of seafood contaminated with STX is responsible for paralytic shellfish poisoning (PSP). Humans are among the species most sensitive to PSP; neurological symptoms of exposure range from tingling of the extremities to severe paralysis. The objective of this study was to determine the effects of STX exposure on developmental processes during early embryogenesis. This study was designed to test the hypothesis that early developmental exposure to STX would disrupt key processes, particularly those related to neural development. Zebrafish embryos were exposed to STX (24 or 48 pg) or vehicle (0.3 mM HCl) at 6 h post fertilization (hpf) via microinjection. There was no overt toxicity but starting at 36 hpf there was a temporary lack of pigmentation in STX-injected embryos, which resolved by 72 hpf. Using high performance liquid chromatography, we found that STX was retained in embryos up to 72 hpf in a dose-dependent manner. Temporal transcriptional profiling of embryos exposed to 48 pg STX per embryo revealed no differentially expressed genes (DEGs) at 24 hpf, but at 36 and 48 hpf, there were 3547 and 3356 DEGs, respectively. KEGG pathway analysis revealed significant enrichment of genes related to focal adhesion, adherens junction and regulation of actin cytoskeleton, suggesting that cell-cell and cell-extracellular matrix interactions were affected by STX. Genes affected are critical for axonal growth and the development of functional neural networks. We confirmed these findings by visualizing axonal defects in transgenic zebrafish with fluorescently labeled sensory neurons. In addition, our gene expression results suggest that STX exposure affects both canonical and noncanonical functions of VGSCs. Given the fundamental role of VGSCs in both physiology and development, these findings offer valuable insights into effects of exposure to neurotoxins.

A diversity of chemicals are intentionally added to plastics to enhance their properties and aid in manufacture. Yet, the accumulated chemical composition of these materials is essentially unknown even to those within the supply chain, let alone to consumers or recyclers. Recent legislated and voluntary commitments to increase recycled content in plastic products highlight the practical challenges wrought by these chemical mixtures, amid growing public concern about the impacts of plastic-associated chemicals on environmental and human health. In this Perspective, we offer guidance for plastics manufacturers to collaborate across sectors and critically assess their use of added chemicals. The ultimate goal is to use fewer and better additives to promote a circular plastics economy with minimal risk to humans and the environment.

Microplastics and nanoplastics are found in marine biota across a wide range of trophic levels and environments. While a large portion of the information about plastic exposure comes from gastrointestinal (GI) data, the relevance of particle accumulation from an oral exposure compared with other types of exposure (e.g. dermal, respiratory) is unknown. To address this gap in knowledge, larval zebrafish (7 days post fertilization) were exposed to two different sizes of nanoplastics through either oral gavage or a waterborne exposure. Larvae were tracked for 48 h post exposure (hpe) to assess the migration and elimination of plastics. Larvae eliminated orally gavaged nanoplastics within 48 hpe. Oral gavage showed limited particle movement from the GI tract into other tissues. In contrast, waterborne nanoplastic-exposed larvae displayed notable fluorescence in tissues outside of the GI tract. The 50 nm waterborne-exposed larvae retained the particles past 48 hpe, and showed accumulation with neuromasts. For both sizes of plastic particles, the nanoplastics were eliminated from non-GI tract tissues by 24 hpe. Our results suggest that waterborne exposure leads to greater accumulation of plastic in comparison to oral exposure, suggesting that plastic accumulation in certain tissues is greater via routes of exposure other than oral consumption.

Popularized on social media, hand-moldable plastics are formed by consumers into tools, trinkets, and dental prosthetics. Despite the anticipated dermal and oral contact, manufacturers share little information with consumers about these materials. Inherent to their function, moldable plastics pose a risk of dermal and oral exposure to unknown leachable substances. We analyzed 12 moldable plastics advertised for modeling and dental applications and determined them to be polycaprolactone (PCL) or thermoplastic polyurethane (TPU). The bioactivities of the most popular brands advertised for modeling applications of each type of polymer were evaluated using a zebrafish embryo bioassay. Both products were sold as microplastic-sized resin pellets. While water-borne exposure to the TPU pellets did not affect the targeted developmental endpoints at any concentration tested, the PCL pellets were acutely toxic above 1 pellet/mL. Aqueous leachates of the PCL pellets demonstrated similar toxicity. Methanolic extracts from the PCL pellets were assayed for their bioactivity using the Attagene FACTORIAL platform. Of the 69 measured endpoints, the extracts activated nuclear receptors and transcription factors for xenobiotic metabolism (pregnane X receptor, PXR), lipid metabolism (peroxisome proliferator-activated receptor gamma, PPARg), and oxidative stress (nuclear factor erythroid 2-related factor 2, NRF2). By non-targeted high-resolution comprehensive two-dimensional gas chromatography (GC×GC-HRT), we tentatively identified several compounds in the methanolic extracts, including PCL oligomers, a phenolic antioxidant, and residues of suspected anti-hydrolysis and crosslinking additives. In a follow-up zebrafish embryo bioassay, because of its stated high purity, biomedical grade PCL was tested to mitigate any confounding effects due to chemical additives in the PCL pellets; it elicited comparable acute toxicity. From these orthogonal and complementary experiments, we suggest that the toxicity was due to oligomers and nanoplastics released from the PCL rather than chemical additives. These results challenge the perceived and assumed inertness of plastics and highlight their multiple sources of toxicity.

Many roles of aryl hydrocarbon receptor repressor in intestinal immune homeostasis are discussed.

While plastic pollution threatens ecosystems and human health, the use of plastic products continues to increase. Limiting its harm requires design strategies for plastic products informed by the threats that plastics pose to the environment. Thus, we developed a sustainability metric for the ecodesign of plastic products with low environmental persistence and uncompromised performance. To do this, we integrated the environmental degradation rate of plastic into established material selection strategies, deriving material indices for environmental persistence. By comparing indices for the environmental impact of on-the-market plastics and proposed alternatives, we show that accounting for the environmental persistence of plastics in design could translate to societal benefits of hundreds of millions of dollars for a single consumer product. Our analysis identifies the materials and their properties that deserve development, adoption, and investment to create functional and less environmentally impactful plastic products.

In May 2021, the M/V X-Press Pearl ship fire disaster led to the largest maritime spill of resin pellets (nurdles) and burnt plastic (pyroplastic). Field samples collected from beaches in Sri Lanka nearest to the ship comprised nurdles and pieces of pyroplastic. Three years later, the toxicity of the spilled material remains unresolved. To begin understanding its potential toxicity, solvent extracts of the nurdles and pyroplastic were screened for their bioactivity by several Attagene FACTORIAL bioassays (TF, NR, and AquaTox), which measured the activity of a combined 70 human transcription factor response elements and nuclear receptors and 6 to 7 nuclear receptors for each of three phylogenetically distinct fish species. Extracts of the pyroplastics robustly activated end points for the human aryl hydrocarbon receptor (AhR), estrogen receptor (ER), pregnane X receptor (PXR), peroxisome proliferator-activated receptor (PPAR), retinoid X receptor (RXR), and oxidative stress (NRF2) and had the potential for activation of several others. The bioactivity profile of the pyroplastics was most similar (similarity score = 0.96) to that of probable human carcinogens benzo[b]fluoranthene and benzo[k]fluoranthene despite the extracts being a complex mixture of thousands of compounds. The activity diminished only slightly for extracts of pyroplastic collected eight months after the spill. The AquaTox FACTORIAL bioassay measured the activation of ERα, ERβ, androgen receptor (AR), PPARα, PPARγ, and RXRβ for human, zebrafish (Danio rerio), Japanese medaka (Oryzias latipes), and rainbow trout (Oncorhynchus mykiss), revealing species-specific sensitivities to the chemicals associated with the pyroplastics. These findings provide needed information to guide long-term monitoring efforts, make hazard assessments of the spilled material, and direct further research on pyroplastic, an emerging global contaminant.

[This corrects the article DOI: 10.5334/aogh.4056.].

In May 2021, the M/V X-Press Pearl container ship burned for 2 weeks, leading to the largest maritime spill of resin pellets (nurdles). The disaster was exacerbated by the leakage of other cargo and the ship's underway fuel. This disaster affords the unique opportunity to study a time-stamped, geolocated release of plastic under real-world conditions. Field samples collected from beaches in Sri Lanka nearest to the ship comprised nurdles exposed to heat and combustion, burnt plastic pieces (pyroplastic), and oil-plastic agglomerates (petroplastic). An unresolved question is whether the 1600+ tons of spilled and recovered plastic should be considered hazardous waste. Due to the known formation and toxicity of combustion-derived polycyclic aromatic hydrocarbons (PAHs), we measured 20 parent and 21 alkylated PAHs associated with several types of spilled plastic. The maximum PAH content of the sampled pyroplastic had the greatest amount of PAHs recorded for marine plastic debris (199,000 ng/g). In contrast, the sampled unburnt white nurdles had two orders of magnitude less PAH content. The PAH composition varied between the types of spilled plastic and presented features typical of and conflicting with petrogenic and pyrogenic sources. Nevertheless, specific markers and compositional changes for burning plastics were identified, revealing that the fire was the main source of PAHs. Eight months after the spill, the PAH contents of sampled stray nurdles and pyroplastic were reduced by more than 50%. Due to their PAH content exceeding levels allowable for plastic consumer goods, classifying burnt plastic as hazardous waste may be warranted. Following a largely successful cleanup, we recommend that the Sri Lankans re-evaluate the identification, handling, and disposal of the plastic debris collected from beaches and the potential exposure of responders and the public to PAHs from handling it. The maritime disaster underscores pyroplastic as a type of plastic pollution that has yet to be fully explored, despite the pervasiveness of intentional and unintentional burning of plastic globally.

Harmful algal blooms (HABs) produce neurotoxins that affect human health. Developmental exposure of zebrafish embryos to the HAB toxin domoic acid (DomA) causes myelin defects, loss of reticulospinal neurons, and behavioral deficits. However, it is unclear whether DomA primarily targets myelin sheaths, leading to the loss of reticulospinal neurons, or reticulospinal neurons, causing myelin defects. Here, we show that while exposure to DomA at 2 dpf did not reduce the number of oligodendrocyte precursors prior to myelination, it led to fewer myelinating oligodendrocytes that produced shorter myelin sheaths and aberrantly wrapped neuron cell bodies. DomA-exposed larvae lacked Mauthner neurons prior to the onset of myelination, suggesting that axonal loss is not secondary to myelin defects. The loss of the axonal targets may have led oligodendrocytes to inappropriately myelinate neuronal cell bodies. Consistent with this, GANT61, a GLI1/2 inhibitor that reduces oligodendrocyte number, caused a reduction in aberrantly myelinated neuron cell bodies in DomA-exposed fish. Together, these results suggest that DomA initially alters reticulospinal neurons and the loss of axons causes aberrant myelination of nearby cell bodies. The identification of initial targets and perturbed cellular processes provides a mechanistic understanding of how DomA alters neurodevelopment, leading to structural and behavioral phenotypes.

Sunlight transforms plastic into water-soluble products, the potential toxicity of which remains unresolved, particularly for vertebrate animals. We evaluated acute toxicity and gene expression in developing zebrafish larvae after 5 days of exposure to photoproduced (P) and dark (D) leachates from additive-free polyethylene (PE) film and consumer-grade, additive-containing, conventional, and recycled PE bags. Using a "worst-case" scenario, with plastic concentrations exceeding those found in natural waters, we observed no acute toxicity. However, at the molecular level, RNA sequencing revealed differences in the number of differentially expressed genes (DEGs) for each leachate treatment: thousands of genes (5442 P, 577 D) for the additive-free film, tens of genes for the additive-containing conventional bag (14 P, 7 D), and none for the additive-containing recycled bag. Gene ontology enrichment analyses suggested that the additive-free PE leachates disrupted neuromuscular processes via biophysical signaling; this was most pronounced for the photoproduced leachates. We suggest that the fewer DEGs elicited by the leachates from conventional PE bags (and none from recycled bags) could be due to differences in photoproduced leachate composition caused by titanium dioxide-catalyzed reactions not present in the additive-free PE. This work demonstrates that the potential toxicity of plastic photoproducts can be product formulation-specific.

ADVERTISEMENT RETURN TO ISSUEViewpointNEXTBiomaterials Science Can Offer a Valuable Second Opinion on Nature's Plastic MaladyBryan D. James*Bryan D. JamesDepartment of Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543, United States*Email: [email protected]More by Bryan D. Jameshttps://orcid.org/0000-0002-6104-8310, Mark E. HahnMark E. HahnDepartment of Biology, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543, United StatesMore by Mark E. Hahnhttps://orcid.org/0000-0003-4358-2082, and Christopher M. ReddyChristopher M. ReddyDepartment of Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543, United StatesMore by Christopher M. Reddyhttps://orcid.org/0000-0002-7814-2071Cite this: Environ. Sci. Technol. 2022, 56, 3, 1475–1477Publication Date (Web):January 7, 2022Publication History Received6 November 2021Published online7 January 2022Published inissue 1 February 2022https://pubs.acs.org/doi/10.1021/acs.est.1c07569https://doi.org/10.1021/acs.est.1c07569article-commentaryACS PublicationsCopyright © 2022 American Chemical Society. This publication is available under these Terms of Use. Request reuse permissions This publication is free to access through this site. Learn MoreArticle Views3992Altmetric-Citations3LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail PDF (1 MB) Get e-Alertsclose Get e-Alerts

Genome-wide association studies have identified risk loci linked to inflammatory bowel disease (IBD)1—a complex chronic inflammatory disorder of the gastrointestinal tract. The increasing prevalence of IBD in industrialized countries and the augmented disease risk observed in migrants who move into areas of higher disease prevalence suggest that environmental factors are also important determinants of IBD susceptibility and severity2. However, the identification of environmental factors relevant to IBD and the mechanisms by which they influence disease has been hampered by the lack of platforms for their systematic investigation. Here we describe an integrated systems approach, combining publicly available databases, zebrafish chemical screens, machine learning and mouse preclinical models to identify environmental factors that control intestinal inflammation. This approach established that the herbicide propyzamide increases inflammation in the small and large intestine. Moreover, we show that an AHR–NF-κB–C/EBPβ signalling axis operates in T cells and dendritic cells to promote intestinal inflammation, and is targeted by propyzamide. In conclusion, we developed a pipeline for the identification of environmental factors and mechanisms of pathogenesis in IBD and, potentially, other inflammatory diseases. The herbicide propyzamide increases inflammation in the small and large intestine, and the AHR–NF-κB–C/EBPβ signalling axis—which operates in T cells and dendritic cells to promote intestinal inflammation—is targeted by propyzamide.