Environmental occurrence, biological effects, and health implications of zinc pyrithione: A review.

Due to the detrimental effects on aquatic organisms and ecosystem, tributyltin as a antifouling agent have been banned worldwide since 1990s. As a replacement for tributyltin, zinc pyrithione (ZnPT) has emerged as a new environmentally friendly antifouling agent. However, the widespread use of ZnPT unavoidably leads to the occurrence and accumulation in aquatic environments, especially in waters with limited sunlight. Despite empirical evidence demonstrating the ecotoxicity and health risks of ZnPT to different organisms, there has been no attempt to compile and interpret this data. The present review revealed that over the past 50 years, numerous studies have documented the toxicity of ZnPT in various organisms, both in vitro and in vivo. However, long-term effects and underlying mechanisms of ZnPT on biota, particularly at environmentally realistic exposure levels, remain largely unexplored. In-depth studies are thus necessary to generate detailed ecotoxicological information of ZnPT for environmental risk assessment and management.

Toxicological effects of tire rubber-derived 6PPD-quinone, a species-specific toxicant, and dithiobisbenzanilide (DTBBA) in the marine rotifer Brachionus koreanus.

The ingredients of tire-rubber products include a complex range of chemicals additives, most of which are leached into surrounding water as unmeasured toxicants with unexplored ecotoxicological impacts. The present study summarizes the reported species-specific acute toxicity of N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine-quinone (6PPD-Q), the ozonation product of anti-oxidant 6PPD used in tire rubber. Also, chronic toxicity and oxidative response of 6PPD-Q and another tire-rubber derivative, 2',2'''-dithiobisbenzanilide (DTBBA), in rotifer Brachionus koreanus were investigated. Although 6PPD-Q has been reported to be highly toxic to several species of salmonids, only moderate chronic toxicity was observed in B. koreanus. In contrast, DTBBA significantly retarded the population growth and fecundity. The varying toxicity of 6PPD-Q and DTBBA was linked to the level of reactive oxygen species in which DTBBA exposure caused a significant concentration-dependent increase. Our results imply unanticipated risks to aquatic species posed by chemical additives in tire-rubber which may be considered emerging contaminants of toxicological concern.

Environmental DNA surveillance of biocontamination in a drinking water treatment plant.

A clean and adequate supply of drinking water is essential to life and good health. However, despite the risk of biologically derived contamination of drinking water, monitoring of invertebrate outbreaks has relied primarily on naked-eye inspections that are prone to errors. In this study, we applied environmental DNA (eDNA) metabarcoding as a biomonitoring tool at seven different stages of drinking water treatment, from prefiltration to release from household faucets. While the composition of invertebrate eDNA communities reflected the communities of the source water in earlier stages of the treatment, several predominant invertebrate taxa (e.g., rotifer) were shown to be introduced during purification, but most were eliminated in later treatment stages. In addition, the limit of detection/quantification of PCR assay and read capacity of high-throughput sequencing was assessed with further microcosm experiments to estimate the applicability eDNA metabarcoding to the biocontamination surveillance in drinking water treatment plants (DWTPs). Here we propose a novel eDNA-based approach for sensitive and efficient surveillance of invertebrate outbreaks in DWTPs.

Maternal exposure to nanoplastic induces transgenerational toxicity in the offspring of rotifer Brachionus koreanus.

Nanoplastics have received a great deal of attention as evidence of their potential harmful effects on aquatic biota. In zooplankton, ingestion is known as a major uptake route of nanoplastics due to the low feeding selectivity of filter-feeding organisms. In this study, we propose maternal transfer as an alternative uptake route of nanoplastic in the rotifer Brachionus koreanus. Exposure to nanoplastics in parental rotifers induced maternal transfer in offspring, as revealed by fluorescence in rotifer eggs. We further verified that egg shells are not permeable to nanoplastics, and the observed fluorescence was associated with nanoplastic particles, not leached fluorescent dye, supporting the idea that nanoplastics can be transferred through an intrinsic maternal transfer route. This maternal transfer induced adverse effects on life-cycle parameters, including development and reproduction, in offspring rotifers, and was associated with oxidative stress. The results of this study shed light on the ecological impacts of nanoplastics in marine environments.

Short- and long-term single and combined effects of microplastics and chromium on the freshwater water flea Daphnia magna.

In this study, we investigated the individual and combined effects of microplastics (MPs) and chromium (Cr) on the freshwater water flea Daphnia magna by measuring mortality, bioaccumulation, antioxidative response, multixenobiotic resistance activity, and sestrin-related mitochondrial biogenesis in short-term assays and in vivo endpoints including reproduction and adult survival rate in long-term assays. Exposure to MPs, Cr, and their combination caused significant deleterious effects and acute toxicity in D. magna. Alterations in oxidative stress occurred in the groups treated with MPs and Cr alone and together. However, upon co-exposure to MPs, the Cr concentration, measured by inductively coupled plasma optical emission spectroscopy, decreased, suggesting that MPs and Cr interact with each other. Based on enzymatic activities, we noted a decrease in MP egestion via inhibition of P-glycoprotein activity in the MP-exposed groups, and multidrug resistance-associated protein activity increased in some of the MP-exposed animals depending on Cr concentration. On the other hand, MP exposure seemed to lead to mitochondrial transcription dysfunction induced by Cr via sestrin-related mitochondrial biogenesis. Overall, these results indicate that co-exposure to MPs and Cr causes acute toxicity in D. magna but lacks the chronic toxicity (21 days) and mitochondrial dysfunction caused by Cr exposure alone.

The single and combined effects of mercury and polystyrene plastic beads on antioxidant-related systems in the brackish water flea: toxicological interaction depending on mercury species and plastic bead size.

Plastics are considered as a major threat to marine environments owing their high usage, persistence, and negative effects on aquatic organisms. Although they often exist as mixtures in combination with other pollutants (e.g., mercury (Hg)) in aquatic ecosystems, the combined effects of plastics and ambient pollutants remain unclear. Therefore, in the present study, we investigated the toxicological interactions between Hg and plastics using two Hg species (HgCl2 and MeHgCl) and different-sized polystyrene (PS) beads (diameter: 0.05, 0.5, and 6-µm) in the brackish water flea Diaphanosoma celebensis. The single and combined effects of Hg and PS beads on mortality were investigated, and changes in the antioxidant system and lipid peroxidation were further analyzed. After 48-h exposure to single Hg, HgCl2 induced a higher mortality rate than MeHgCl. The combined exposure test showed that 0.05-µm PS beads can enhance the toxicity of both the Hg species. The expression of GST-mu, glutathione S-transferease (GST) activity and malondialdehyde (MDA) content increased significantly after exposure to Hg alone (HgCl2 or MeHgCl) exposure. Combined exposure with PS beads modulated the effects of Hg on the antioxidant system depending on bead size and the Hg species. In particular, the 0.05-µm beads significantly increased the expression level of GST-mu, GST activity and MDA content, regardless of Hg species. These findings suggest that toxicological interactions between Hg and PS beads depend on the type of Hg species and the size of PS beads; nano-sized 0.05-µm PS beads can induce synergistic toxicity with Hg.

Contrasting toxicity of polystyrene nanoplastics to the rotifer Brachionus koreanus in the presence of zinc oxide nanoparticles and zinc ions.

Emerging contaminants such as nanoplastics and nanoparticles likely experience similar environmental behaviours, fate and effects but our knowledge of their combined toxicity is scanty. This study, therefore, investigated the joint toxicity of polystyrene nanoplastics (PNPs) and zinc oxide nanoparticles (ZnO-NPs) to an ecologically important rotifer Brachionus koreanus, and compared with the joint toxicity of PNPs and Zn ions (Zn-IONs from ZnSO4·7H2O). With increasing concentration, ZnO-NPs formed significant agglomeration with PNPs for up to 1.3 times of the original hydrodynamic size of ZnO-NPs, alongside doubling in their sedimentation and thereby losing 58% of their released Zn ions. In contrast, the availability of Zn-IONs was less affected by the agglomeration and sedimentation of PNPs, with only a loss of 18% of Zn ions at the highest concentration of PNPs. Consequently, as suggested by Concentration Addition and Independent Action models and the Model Deviation Ratios, ZnO-NPs and PNPs exerted an antagonistic interaction whereas Zn-IONs and PNPs exhibited an additive effect. We also advocate the use of the Nonparametric Response Surface method, which is more useful to predict the toxicity of chemical mixtures with interacting effects. Our findings suggested a potential difference between particle-particle and particle-ion interactions, especially at higher test concentrations, which may eventually affect their toxicity. We, therefore, call for a more systematic evaluation of commonly coexisting chemical mixtures which consist of nanoplastics and manufactured nanomaterials.

Modulation of ecdysteroid and juvenile hormone signaling pathways by bisphenol analogues and polystyrene beads in the brackish water flea Diaphanosoma celebensis.

Owing to its high production and world-wide usage, plastic pollution is an increasing concern in marine environments. Plastic is decomposed into nano- and micro-sized debris, which negative affect reproduction and development in aquatic organisms. Bisphenol A (BPA), an additive of plastic, is released into the water column upon plastic degradation, and is known as a representative endocrine-disrupting chemical. However, the reproductive effects of plastics and bisphenols at the molecular level have not yet been explored in small marine crustaceans. In this study, we investigated the effects of polystyrene (PS) beads (0.05, 0.5, and 6 - µm) and bisphenol analogues (BPs; BPA, BPS, and BPF) on reproduction and development of small marine crustaceans. Effects on transcriptional changes in ecdysteroid and juvenile hormone (JH) signaling pathway-related genes were examined in the brackish water flea Diaphanosoma celebensis exposed to PS beads and BPs for 48 h. As results, BPs and PS beads delayed emergence time of first offspring, and increased fecundity in a concentration-dependent manner. BPs differentially modulated the expression of ecdysteroid and JH signaling pathway-related genes, indicating that BP analogs can disrupt endocrine systems via mechanisms different from those of BPA. PS beads was also changed the gene expression of both pathway, depending on their size and concentration. Our findings suggest that BP analogues and PS beads disrupt the endocrine system by modulating the hormonal pathways, affecting reproduction negatively. This study provides a better understanding of the molecular mode of action of BPs and PS beads in the reproduction of small crustaceans.

Metabolism deficiency and oxidative stress induced by plastic particles in the rotifer Brachionus plicatilis: Common and distinct phenotypic and transcriptomic responses to nano- and microplastics.

Growing experimental data on the adverse effects of microplastic pollution on marine biota indicate that the size of the plastic particles is a key determinant of toxicity. Here, we investigated size-dependent toxicity at different levels of biological organizations in the marine rotifer Brachionus plicatilis, from bioaccumulation as an initiating event to adverse in-vivo outcomes, with ecotoxicogenomic approach to elucidate the size-dependent toxicity of microplastics. Nanoplastics strongly retarded the reproduction and population growth of B. plicatilis, while microplastics were associated with moderate effects. This size dependency could be attributed to the selective induction of oxidative stress by nanoplastic exposure in addition to a metabolic deficiency, which was a common toxicity mechanism with both nano- and microplastic exposure as predicted by transcriptomic analysis. Our findings suggested that metabolic deficiency is a shared toxicity mechanism of nano- and microplastics, while oxidative stress might be responsible for the stronger toxicity of nanoplastics.

Phenotypic toxicity, oxidative response, and transcriptomic deregulation of the rotifer Brachionus plicatilis exposed to a toxic cocktail of tire-wear particle leachate.

Tire-wear particles (TWPs) are potential source of microplastic (MP) pollution in marine environments. Although the hazardous effects of MPs on marine biota have received considerable attention, the toxicity of TWPs and associated leachates remain poorly understood. Here, to assess the toxicity of TWP leachate and the underlying mechanisms of toxicity, the phenotypic and transcriptomic responses of the rotifer Brachionus plicatilis were assessed with chemistry analysis of a TWP leachate. Although acute toxicity was induced, and a variety of metals and polyaromatic hydrocarbons were detected in the leachate, levels were below the threshold for acute toxicity. The results of particle analysis suggest that the acute toxicity observed in our study is the result of a toxic cocktail of micro- and/or nano-sized TWPs and other additives in TWP leachate. The adverse effects of TWP leachate were associated with differential expression of genes related to cellular processes, stress response, and impaired metabolism, with further oxidative stress responses. Our results imply that TWPs pose a greater threat to marine biota than other plastic particles as they constitute a major source of nano- and microplastics that have synergistic effects with the additives contained in TWP leachate.

Effects of atrazine and diuron on life parameters, antioxidant response, and multixenobiotic resistance in non-targeted marine zooplankton.

Atrazine and diuron are among the most widely used antifoulant biocides in the world. Due to their persistence in the environment, they can induce adverse effects on non-targeted organisms. In this study, we investigated the chronic in vivo toxicity of atrazine and diuron with further assessments on oxidative stress responses (e.g., oxidative stress, antioxidant) and multixenobiotic resistance (MXR) function in the rotifer Brachionus koreanus, a non-targeted microzooplanktonic grazer at the primary level of the marine food chain. Although similar oxidative response was shown by both biocides, diuron induced stronger retardation on reproduction and population growth rates of B. koreanus while moderate effects were observed by atrazine. This higher toxicity of diuron was shown to be associated with its stronger inhibition of MXR conferred by P-glycoprotein and multidrug resistance proteins which play as a first line of defense by transporting various toxicants out of a cell. Our study provides new insight into non-targeted effects of biocides on marine zooplankton and mechanisms beyond their different degrees of toxicity.

Phenotypic and transcriptomic responses of the rotifer Brachionus koreanus by single and combined exposures to nano-sized microplastics and water-accommodated fractions of crude oil.

Sorption of organic pollutants on microplastics can be an alternative uptake route for organic pollutants in aquatic organisms. To assess the combined effects of microplastics and organic pollutants, we employed phenotypic and transcriptomic analyses to the responses of the marine rotifer Brachionus koreanus to environmentally relevant concentrations of nano-sized microplastic (0.05 µm), water-accommodated fractions of crude oil, and binary mixtures thereof. Our multigenerational in vivo experiments revealed more than additive effects on population growth of B. koreanus in response to combined exposure, while a single exposure to nano-sized microplastic did not induce observable adverse effects. Synergistic transcriptome deregulation was consistently associated with dramatically higher numbers of differentially expressed genes, and increased gene expression was associated with combined exposure. The majority of synergistic transcriptional alteration was related to metabolism and transcription, with impaired reproduction resulting from energetic reallocation toward adaptation. As further supported by chemistry analysis for polycyclic aromatic hydrocarbons sorption on microplastic, our findings imply that nano-sized microplastics can synergistically mediate the effects of organic pollutants in aquatic organisms.

Effects of polystyrene in the brackish water flea Diaphanosoma celebensis: Size-dependent acute toxicity, ingestion, egestion, and antioxidant response.

Owing to the increasing usage of plastics, their debris is continuously deposited in marine environments, resulting in deleterious effects on aquatic organisms. Although it is known that microplastics disturb the cellular redox status, knowledge of molecular in marine cladocerans is still lacking. In the present study, we investigated the acute toxicity of different-sized polystyrene (PS) beads (0.05, 0.5, and 6-µm diameter), ingestion and egestion patterns, their distribution in the tissues, and their effects on the antioxidant systems in the brackish water flea Diaphanosoma celebensis. All different-sized PS beads showed no mortality at the concentrations used in this study. After 48 h of exposure to PS beads of different sizes, all microbeads were retained in the digestive tract, but the retention time varied according to the bead size. In particular, the group that was exposed to 0.05-µm beads showed widely distributed fluorescence (e.g., in the embryo, and probably in lipid droplets as well as the digestive tract). The transcriptional level and enzyme activities of antioxidants were modulated depending on the size of the PS beads, and lipid peroxidation was induced in groups exposed to 0.05 and 0.5-µm beads. These findings suggest that the size of PS beads is an important factor for cellular toxicity, and can induce size-dependent oxidative stress in this species. This study provides a better understanding of the molecular modes of action of microplastics in marine zooplankton.

Key mechanisms of micro- and nanoplastic (MNP) toxicity across taxonomic groups.

Micro- and nanoplastics (MNPs) are ubiquitous in aquatic and terrestrial environments, and detrimental biological effects have been observed on a variety of organisms, from bacteria and alga to plants and animals. A fast-growing number of toxicological studies report diverse responses and wide species-dependent sensitivity upon MNP exposure. While studies are dominated by in vivo animal tests, our understanding of cellular toxicity and the corresponding toxicity mechanisms is still limited. This challenges the proper assessment of environmental hazards and health risks of MNPs. In this review, we gathered and analyzed the up-to-date studies on humans, animals, plants, alga, and bacteria, and identified the similarities and differences in key toxicity mechanisms of MNPs across different taxonomic groups. Particularly, human cell-based studies at the cellular level provide fundamental and valuable information on the key toxicity mechanisms, which are essential to answer the question of whether and how MNPs pose health threats. In general, toxicity mechanisms of MNPs depend on their size, surface characteristics, polymer type, as well as cell type. Plausible toxicity mechanisms mainly include membrane disruption, extracellular polymeric substance disruption, reactive oxygen species generation, DNA damage, cell pore blockage, lysosome destabilization, and mitochondrial depolarization. A deeper understanding of these key mechanisms in different taxonomic groups can also improve both in vivo and in vitro models useful for predictive impact assessments of plastic pollution on the environment and human health.

The genome of the freshwater monogonont rotifer Brachionus angularis: Identification of phase I, II, and III detoxification genes and their roles in molecular ecotoxicology.

Brachionus spp. rotifers, which are widely distributed in aquatic environments, have been proposed as model organisms for ecotoxicological studies. Although the genomes of several rotifers belonging to the genus Brachionus have been assembled, the genome for the freshwater rotifer Brachionus angularis remains unknown. In this study, we analyzed the whole-genome sequence of B. angularis, which revealed a total length of 56.5 Mb and 21 contigs. The N50 and the GC content were 5.42 Mb and 23.66%, respectively. A total of 13,952 genes were predicted. Of them, we identified the main detoxification-related gene families, including those for cytochrome P450, glutathione S-transferase (GST), and the ATP-binding cassette transporter. In comparison with other Brachionus rotifers, massive species-specific expansion in GST sigma genes was found in B. angularis. This whole-genome analysis of B. angularis provides a basis for molecular ecotoxicological studies and provides useful biological tools for comparative studies of the evolution of detoxification mechanisms in Brachionus spp.

The genome of the European estuarine calanoid copepod Eurytemora affinis: Potential use in molecular ecotoxicology.

In this study, we sequenced and assembled the genome of a European estuarine calanoid copepod using Oxford Nanopore PromethION and Illumina HiSeq 2500 platforms. The length of the assembled genome was 776.1 Mb with N50 = 474.9 kb (BUSCO 85.9%), and the genome consisted of 2473 contigs. A total of 18,014 genes were annotated and orthologous gene clusters were analyzed in comparison to other copepods. In addition, genome-wide identification of cytochrome P450s, glutathione S-transferases, and ATP-binding cassette transporters in E. affinis was performed to determine gene repertoire of these detoxification-related gene families. Results revealed the presence of species-specific gene inventories, indicating that these gene families have evolved through species-specific gene loss/expansion processes, possibly due to adaptation to different environmental stressors. Our study provides a new inventory of the European estuarine calanoid copepod E. affinis genome with emphasis on phase I, II, and III detoxification systems.

Arsenic exposure combined with nano- or microplastic induces different effects in the marine rotifer Brachionus plicatilis.

Besides the adverse biological effects induced by microplastics (MPs), the effects associated with sorption of ambient pollutants on MPs are considered as an emerging environmental problem as MPs act as a mediator of pollutants. The present study examines the combined effects of nano(micro)plastics (NMPs) and arsenic (As) by exposing the marine rotifer Brachionus plicatilis to MP particles at the micro-scale (6 µm) and nano-scale (nanoplastics, NPs) (50 nm) along with As. In vivo toxicity, bioaccumulation, and biochemical reactions were used to examine the effects of combined exposure. The results of in vivo experiments showed that As toxicity increased with NP exposure, whereas toxicity was alleviated by MPs, indicating a different mode of action between NPs and MPs in combination with As. The highest level of As bioaccumulation was detected in NP + As groups, and followed by MP + As and As-only exposure groups, whereas no significant difference between groups was shown for As metabolites. In addition, the activity of several ATP-binding cassette proteins that confer multixenobiotic resistance, which is responsible for efflux of As, was activated by As but significantly inhibited by NP exposure, supporting the findings of in vivo experiments. Our results show that the effects of combining exposure to As with NP and MPs differ depending on particle size and provide an in-depth understanding of both environmental pollutants.

Multigenerational Mitigating Effects of Ocean Acidification on In Vivo Endpoints, Antioxidant Defense, DNA Damage Response, and Epigenetic Modification in an Asexual Monogonont Rotifer.

Ocean acidification (OA) is caused by changes in ocean carbon chemistry due to increased atmospheric pCO2 and is predicted to have deleterious effects on marine ecosystems. While the potential impacts of OA on many marine species have been studied, the multigenerational effects on asexual organisms remain unknown. We found that low seawater pH induced oxidative stress and DNA damage, decreasing growth rates, fecundity, and lifespans in the parental generation, whereas deleterious effects on in vivo endpoints in F1 and F2 offspring were less evident. The findings suggest that multigenerational adaptive effects play a role in antioxidant abilities and other defense mechanisms. OA-induced DNA damage, including double-strand breaks (DSBs), was fully repaired in F1 offspring of parents exposed to OA for 7 days, indicating that an adaptation mechanism may be the major driving force behind multigenerational adaptive effects. Analysis of epigenetic modification in response to OA involved examination of histone modification of DNA repair genes and a chromatin immunoprecipitation assay, as Bombus koreanus has no methylation pattern for CpG in its genome. We conclude that DSBs, DNA repair, and histone modification play important roles in multigenerational plasticity in response to OA in an asexual monogonont rotifer.

Transgenerational acclimation to changes in ocean acidification in marine invertebrates.

The rapid pace of increasing oceanic acidity poses a major threat to the fitness of the marine ecosystem, as well as the buffering capacity of the oceans. Disruption in chemical equilibrium in the ocean leads to decreased carbonate ion precipitation, resulting in calcium carbonate saturation. If these trends continue, calcifying invertebrates will experience difficultly maintaining their calcium carbonate exoskeleton and shells. Because malfunction of exoskeleton formation by calcifiers in response to ocean acidification (OA) will have non-canonical biological cascading results in the marine ecosystem, many studies have investigated the direct and indirect consequences of OA on ecosystem- and physiology-related traits of marine invertebrates. Considering that evolutionary adaptation to OA depends on the duration of OA effects, long-term exposure to OA stress over multi-generations may result in adaptive mechanisms that increase the potential fitness of marine invertebrates in response to OA. Transgenerational studies have the potential to elucidate the roles of acclimation, carryover effects, and evolutionary adaptation within and over generations in response to OA. In particular, understanding mechanisms of transgenerational responses (e.g., antioxidant responses, metabolic changes, epigenetic reprogramming) to changes in OA will enhance our understanding of marine invertebrate in response to rapid climate change.

Generation of albino via SLC45a2 gene targeting by CRISPR/Cas9 in the marine medaka Oryzias melastigma.

To produce albinism in the marine medaka Oryzias melastigma, we disrupted the solute carrier family 45 (SLC45a2) gene by clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 with a single guide RNA (sgRNA). Selected sgRNAs were able to target a SLC45a2 gene as confirmed by genotyping and heteroduplex mobility assay (HMA). Of the survived embryos after injection, 54.2% and 60.0% embryos exhibited albinism phenotype by sgRNA1 and sgRNA2, respectively. Deep sequencing at the on-target sites showed different insertion and deletion (indel) mutation profiles near the DNA cleavage sites, indicating high efficacy of producing SLC45a2 knock-out mutants by this method. Moreover, HMA at the potential off-target sites revealed that off-target activity would be induced at a low rate, or not induced at all. This albino marine medaka will be a good model for marine molecular ecotoxicology in establishment of diverse in vivo endpoints, and the application of this efficient gene targeting method in the marine medaka would be useful tool for mechanistic approaches.