Rapid Mass Conversion for Environmental Microplastics of Diverse Shapes.

Rivers have been recognized as the primary conveyors of microplastics to the oceans, and seaward transport flux of riverine microplastics is an issue of global attention. However, there is a significant discrepancy in how microplastic concentration is expressed in field occurrence investigations (number concentration) and in mass flux (mass concentration). Of urgent need is to establish efficient conversion models to correlate these two important paradigms. Here, we first established an abundant environmental microplastic dataset and then employed a deep neural residual network (ResNet50) to successfully separate microplastics into fiber, fragment, and pellet shapes with 92.67% accuracy. We also used the circularity (C) parameter to represent the surface shape alteration of pellet-shaped microplastics, which always have a more uneven surface than other shapes. Furthermore, we added thickness information to two-dimensional images, which has been ignored by most prior research because labor-intensive processes were required. Eventually, a set of accurate models for microplastic mass conversion was developed, with absolute estimation errors of 7.1, 3.1, 0.2, and 0.9% for pellet (0.50 ≤ C < 0.75), pellet (0.75 ≤ C ≤ 1.00), fiber, and fragment microplastics, respectively; environmental samples have validated that this set is significantly faster (saves ∼2 h/100 MPs) and less biased (7-fold lower estimation errors) compared to previous empirical models.

Non-negligible Toxicity to Fish in the Early Life Stages Triggered by Aqueous Leachate of Takeaway Plastic Containers.

Ordering takeout is a growing social phenomenon and may raise public health concerns. However, the associated health risk of compounds leaching from plastic packaging is unknown due to the lack of chemical and toxicity data. In this study, 20 chemical candidates were tentatively identified in the environmentally relevant leachate from plastic containers through the nontargeted chemical analysis. Three main components with high responses and/or predicted toxicity were further verified and quantified, namely, 3,5-di-tert-butyl-4-hydroxycinnamic acid (BHC), 2,4-di-tert-butylphenol (2,4-DTBP), and 9-octadecenamide (oleamide). The toxicity to zebrafish larvae of BHC, a degradation product of a widely used antioxidant Irganox 1010, was quite similar to that of the whole plastic leachate. In the same manner, RNA-seq-based ingenuity analysis showed that the affected canonical pathways of zebrafish larvae were quite comparable between BHC and the whole plastic leachate, i.e., highly relevant to neurological disease, metabolic disease, and even behavioral disorder. Longer-term exposure (35 days) did not cause any effect on adult zebrafish but led to decreased hatching rate and obvious neurotoxicity in zebrafish offspring. Collectively, this study strongly suggests that plastic containers can leach out a suite of compounds causing non-negligible impacts on the early stages of fish via direct or parental exposure.

Diel Pattern of Microplastic Residues in Zebrafish.

As one of the emerging pollutants, microplastics (MPs) can be taken up by aquatic organisms through ingestion. However, little is known about the uptake pattern in organisms over time and the associated mechanisms of retention patterns. The present study aims to elucidate these patterns in fish, their relationship with light/dark conditions, and examine the uptake kinetic process of small-sized plastic pollutants, especially during the long-neglected dark period. Zebrafish were sampled every 2 h during the light and dark periods after exposure to an environmentally relevant concentration (100 items/L) of MPs. The results demonstrated that MP residues in zebrafish decreased during the dark period rather than increased over time. The MP retention rhythm and the swimming behavior of exposed zebrafish displayed a statistically significant light/dark variation. Moreover, a very strong and statistically significant positive correlation was found between the swimming speed of zebrafish and the number of MP residues in the gastrointestinal tracts of zebrafish. These results clearly demonstrate that fibrous MP residues in the fish have a discernible diel pattern. This work improves the understanding of the dynamic residual process of MPs in organisms and calls for further in-depth circadian toxicokinetic studies to better suit particle pollutants.

Crack Patterns of Environmental Plastic Fragments.

Secondary microplastics usually come from the breakdown of larger plastics due to weathering and environmental stress cracking of plastic wastes. In the present study, 5013 plastic fragments were collected from coastal beaches, estuary dikes, and lake banks in China. The fragment sizes ranged from 0.2 to 17.1 cm, and the dominant polymers were polypropylene and polyethylene. Cracks were observed on the surfaces of 49-56% of the fragments. Based on the extracted crack images, we proposed a general crack pattern system including four crack types with specific definitions, abbreviations, and symbols. The two-dimensional spectral analysis of the cracks suggests that the first three patterns showed good regularity and supported the rationality of the pattern system. Some crack metrics (e.g., line density) were closely correlated with the carbonyl index and additives (e.g., phthalate esters) of fragments. For crack investigation in field, we proposed a succinct protocol, in which five crack ranks were established to directly characterize the degree of cracking based on the line density values. The system was successfully applied to distinguish the differences in crack features at two representative sites, which indicates that crack pattern is a useful tool to describe the morphological changes of plastic surfaces in the environment.

Plastic debris in coastal macroalgae.

Macroalgae are the most productive marine macrophytes in the coastal ecosystem. Although plastic debris has been ubiquitously detected in marine animals, little is known about plastic pollution in macroalgae and how they interact with each other. In this study, the occurrence of plastic debris including microplastics was investigated in 5 macroalgae species that are commonly found along the Chinese shorelines. These species consisted of Gracilaria lemaneiformis, Chondrus ocellatus, Ulva lactuca, Ulva prolifera and Saccharina japonica. We categorized the plastic debris into 3 size classes, i.e., macroplastics (>25 mm), mesoplastics (5-25 mm), and microplastics (1 µm-5 mm). It was shown that there were 5 loading patterns of plastic debris interaction with the macroalgae. The 5 patterns included entanglement, adherence, wrapping, embedment, and entrapment by epibionts. According to direct observations through the non-digestion method, all 3 size classes of plastics were found in the macroalgae. The abundances were 0-201.5 (macroplastics), 0-1178.0 (mesoplastics) and 0-355.6 (microplastics) items/kg dry weight, respectively. These plastics were dominated by fibers (52.2%) in shape, 1-5 mm (39.6%) in size, and polystyrene (36.5%) in composition. According to indirect observations through the digestion method, only 2 size classes of plastics were identified in the macroalgae: mesoplastics, 0 to 888.9 items/kg dry weight; microplastics, 148.1 to 5889.0 items/kg dry weight. These plastics were prevailing by fibers (71.5%) in shape, 1-5 mm (52%) in size, and polyester (29.3%) in composition. Furthermore, plastic characteristics in the detected macroalgae were related to their species, sampling regions, and beach types based on the results of similarity and principal component analysis. This study indicated that macroalgae utilized diverse pathways for loading plastics in the coastal environment. Meanwhile, environmental factors significantly influenced the distribution of plastics loaded by macroalgae.

Gonadal Atresia, Estrogen-Responsive, and Apoptosis-Specific mRNA Expression in Marine Mussels from the East China Coast: A Preliminary Study.

This preliminary survey analysed mussel atresia incidences, estrogen-responsive and apoptotic-specific molecular end points, and aqueous and gonadal levels of selected estrogens from the East China coast. Estrogen levels were low (e.g. < LOD-28.36 ng/L, < LOD-3.88 ng/g wet weight of tissue for BPA) relative to worldwide freshwater environments, but high oocyte follicle atresia incidences (up to 26.6%) occurred at selected sites. Expression of estrogen-responsive ER2 was significantly increased in males relative to females at sites with high atresia incidences in females. A second estrogen-responsive gene, V9, was significantly increased at two sites in April in females relative to males; the opposite was true for the remaining two sites. Apoptosis-specific genes (Bcl-2, fas) showed elevated expression in males relative to females at the site with the highest atresia incidence. These results provide coastal estrogen levels and the utility of several estrogen-specific molecular-level markers for marine mussels.

Distribution and translocation of micro- and nanoplastics in fish.

Microplastics (MPs) and nanoplastics (NPs) are regarded as emerging particulate contaminants. Here, we first summarize the distribution of plastic particles in fish. Field investigations verify the presence of various kinds of fibrous, spherical, and fragmentary MPs in fish gastrointestinal tract and gills, and specifically in muscle and liver. Laboratory works demonstrate that NPs even penetrate into blood vessels of fish and pass onto next generations. Second, we systematically discuss the translocation ability of MPs and NPs in fish. MPs can enter early-developing fish through adherence, and enter adult fish internal organs by intestine absorption or epidermis infiltration. NPs can not only penetrate into fish embryo blastopores, but also reach adult fish internal organs through blood circulation. Third, the cellular basis for translocation of plastic particles, NPs in particular, into cells are critically reviewed. Endocytosis and paracellular penetration are two main pathways for them to enter cells and intercellular space, respectively. Finally, we compare the chemical and physical properties among various particular pollutants (MPs, NPs, settleable particulate matters, and manufactured nanomaterials) and their translocation processes at different biological levels. In future studies, it is urgent to break through the bottleneck techniques for NPs quantification in field environmental matrix and organisms, re-confirm the existence of MPs and NPs in field organisms, and develop more detailed translocating mechanisms of MPs and NPs by applying cutting-edge tracking techniques.

Fish Ingest Microplastics Unintentionally.

Microplastics (size of plastic debris <5 mm) occur in various environments worldwide these days and cause detrimental effects on biota. However, the behavioral responses of fish to microplastics in feeding processes are not well understood. In the present study, juveniles from four fish species and two common shapes of microplastics were used to explore fish feeding responses. We found swallowing-feeding fish ingested more pellets than filtering- and sucking-feeding fish. With high-definition and high-speed observational experiments, we found that all species did not actively capture microfibers; instead, they passively sucked in microfibers while breathing. Surprisingly, fish showed a rejective behavior, which was spontaneously coughing up microfibers mixed with mucus. Nevertheless, some of the microfibers were still found in the gastrointestinal tracts and gills of fish, while abundances of ingested microfibers were increased in the presence of food. Our findings reveal a common phenomenon that fish ingest microplastics inadvertently rather than intentionally. We also provide insights into the pathways via which microplastics enter fish and potential strategies to assess future ecological risk and food safety related to microplastics.

Microplastics habituated with biofilm change decabrominated diphenyl ether degradation products and thyroid endocrine toxicity.

Microplastics (MPs) are rapidly colonized by microbial biofilms in a natural aquatic environment, and the nature of the microbial community and type of MP can result in different degradation products of organic pollutants. Here, we quantified the degradation products of a ubiquitously detected pollutant, decabrominated diphenyl ether (BDE-209), under both light-only and biota conditions and in the absence or presence of three kinds of MPs, styrofoam polystyrene, hard polyamide, and polypropylene film. The results showed that the BDE-209 concentration increased by 0.7-2.8 fold in the presence of MPs, probably due to the "sustained release" desorption effect. Under light-only conditions, the penta- and hexa-BDE concentrations in the presence of styrofoam or hard MPs were significantly reduced, which can be deemed a beneficial effect. However, when biota were present, the debromination products increased with the addition of MPs, particularly in the presence of styrofoam MPs. These products caused a 1.7-fold upregulation in triiodothyronine content and a 5.9-fold upregulation of thyroid stimulating hormone ß expression in zebrafish larvae. The increase in debromination products could be attributed to the distinct high abundance of the bacteria Chloroflexi, Proteobacteria, and Basidiomycotina on styrofoam MPs that can participate in pollutant degradation. Collectively, our results indicate that MPs can alter the degradation pathways of BDE-209 and increase the toxicity to the endocrine system and the thyroid in aquatic organisms.

Microplastic Fallout in Different Indoor Environments.

Microplastics in the air have gradually attracted our attention in recent years; however, temporal and spatial trends of microplastics in indoor air are rarely discussed. In the present study, we tracked microplastic fallout in a dormitory, an office, and a corridor on both workdays and weekends for three months. In addition, an air conditioner was used to understand airflow influence on microplastic resuspension in the dorm. Among the three sampling sites, the highest average microplastic abundance appeared in the dormitory (9.9 × 103 MPs/m2/d), followed by the office (1.8 × 103 MPs/m2/d) and the corridor (1.5 × 103 MPs/m2/d). In the dormitory, the average MP abundance on weekends (1.4 × 104 MPs/m2/d) was approximately three times of that on weekdays (5.8 × 103 MPs/m2/d). In the office; however, the abundance on weekends (1.2 × 103 MPs/m2/d) was 50% of that on weekdays (2.4 × 103 MPs/m2/d). Microplastic fallout existed mostly in the form of fibers and showed similar polymer compositions to the textile products used in indoor environments. The airflow tests using an air conditioner suggested that airflow turbulence increased resuspension of microplastics. Taken together, we conclude that indoor environments are prone to serious microplastic pollution, but microplastic level varies greatly due to different characteristics of indoor setting. Our results also indicate that textile quantity is one of the main factors affecting microplastic abundance in indoor air, whereas air conditioner-induced airflow turbulence can cause microplastic migration in indoor environments.

A Review of Microplastics in Table Salt, Drinking Water, and Air: Direct Human Exposure.

The ubiquity of microplastics in aquatic and terrestrial environments and related ecological impacts have gained global attention. Microplastics have been detected in table salt, drinking water, and air, posing inevitable human exposure risk. However, rigorous analytical methods for detection and characterization of microplastics remain scarce. Knowledge about the potential adverse effects on human health via dietary and respiratory exposures is also limited. To address these issues, we reviewed 46 publications concerning abundances, potential sources, and analytical methods of microplastics in table salt, drinking water, and air. We also summarized probable translocation and accumulation pathways of microplastics within human body. Human body burdens of microplastics through table salt, drinking water, and inhalation were estimated to be (0-7.3)×104, (0-4.7)×103, and (0-3.0)×107 items per person per year, respectively. The intake of microplastics via inhalation, especially via indoor air, was much higher than those via other exposure routes. Moreover, microplastics in the air impose threats to both respiratory and digestive systems through breathing and ingestion. Given the lifetime inevitable exposure to microplastics, we urgently call for a better understanding of the potential hazards of microplastics to human health.

Design, Synthesis, Biological Evaluation and Inhibition Mechanism of 3-/4-Alkoxy Phenylethylidenethiosemicarbazides as New, Potent and Safe Tyrosinase Inhibitors.

Tyrosinase plays important roles in many different disease related processes, and the development of its inhibitors is particularly important in biotechnology. In this study, thirty-nine 3-/4-alkoxyphenylethylidenethiosemicarbazides were synthesized as novel tyrosinase inhibitors based on structure-based molecular design. Our experimental results demonstrated that thirty-one of them possess remarkable tyrosinase inhibitory activities with IC50 value below 1 µM, and 5a, 6e, 6g and 6t did not display any toxicity to 293T cell line at the concentration of 1000 µmol/L. According to the inhibitory activities, several compounds were selected for detail investigation on the structure-activity relationships (SARs), mechanisms of enzyme inhibition, inhibitory kinetics and cytotoxicity. In particular, the interaction between the selected inhibitors and the active center of tyrosinase was considered and discussed in detail based on their structural characteristics. Taken together, the results presented here demonstrated that the newly designed compounds are promising candidates for the treatment of tyrosinase-related disorders and further development of them may have significant contribution in biomedical science.

PVC Does Not Influence Cadmium Uptake or Effects in the Mussel (Mytilus edulis).

Microplastics have become a global concern in recent years. In this study, we studied (i) whether the presence of polyvinyl chloride (PVC) microparticles may affect cadmium (Cd) uptake in mussel (Mytilus edulis); and (ii) the biological effects of PVC microparticles exposure alone or in combination with Cd. Significant Cd uptake in digestive gland was observed following Cd exposure. However, PVC did not significantly increase Cd uptake compared with Cd alone treatment. In terms of biological impacts, significantly lower neutral red retention (NRR) time and elevated expression of Metallothionein isoform 20-IV (MT-20) were observed in mussels exposed to Cd alone, or combined with microplastics, yet there was no significant difference between them. catalase (CAT) expression only showed a significant increase in mussels exposed to Cd alone. This work provides an insight into the relationship on resulting biological impacts between these two contaminants.

Microplastics as Both a Sink and a Source of Bisphenol A in the Marine Environment.

Microplastics were demonstrated to be an environmental sink for hydrophobic organic pollutants, while they can also serve as a potential source of such pollutants. In this study, the sorption and release of bisphenol A in marine water were investigated through laboratory experiments. Sorption and desorption isotherms were developed, and the results reveal that sorption and desorption depend on the crystallinity, elasticity, and hydrophobicity of the polymer concerned. The adsorption and partition of bisphenol A can be quantified using a dual-mode model of the sorption mechanisms. Polyamide and polyurethane were found to exhibit the highest sorption capacity for bisphenol A, and it was almost irreversible, probably due to hydrogen bonding. Polyethylenes and polypropylene exhibited high and reversible sorption without noticeable desorption hysteresis. Glassy polystyrene, poly(vinyl chloride), poly(methyl methacrylate), and poly(ethylene terephthalate) exhibited low sorption capacity and only partial reversibility. Low-density polyethylene and polycarbonate microplastic particles were for the first time proved to be a persistent source releasing bisphenol A into aquatic environments. Salinity, pH, coexisting estrogens, and water chemistry influence the sorption/desorption behaviors to different degrees. Plastic particles can serve as transportation vectors for bisphenol A, which may constitute an ecological risk.

Microplastics in Small Waterbodies and Tadpoles from Yangtze River Delta, China.

Although microplastic (MP) pollution in freshwater systems is gaining attention, our knowledge of its distribution in small waterbodies is scarce. Small waterbodies are freshwater habitats to many species, including amphibians, that are vulnerable to MP pollution. This study analyzed the distribution and characteristics of MPs in 25 small waterbodies from the Yangtze River Delta, China. MPs were detected in surface water, sediment, and tadpoles with abundances ranging from 0.48 to 21.52 items L-1, 35.76 to 3185.33 items kg-1, and 0 to 2.73 items individual-1 (0 to 168.48 items g-1), respectively. The dominant shape and polymer of MPs in water and tadpole samples were polyester (PES) fibers, and polypropylene (PP) fibers and fragments were dominant in sediment samples. In addition, MPs were primarily <0.5 mm in length in all samples. Tadpole length was positively correlated to the number of MPs detected. The abundance, shape, and polymer distribution of MPs in tadpoles resembled that of water rather than sediment, suggesting that tadpoles likely take up MPs from the surrounding water. This study demonstrated that MPs are abundant in these small waterbodies and are ingested by resident tadpoles. This may suggest a pathway of MP entry into aquatic and terrestrial food webs.

Single and mixture toxicity of strobilurin and SDHI fungicides to Xenopus tropicalis embryos.

The decline in amphibian populations is a critical threat to global biodiversity, and pesticide pollution is considered as one of the major factors. Although effects of single pesticides on amphibians have been documented, toxicological interactions prevailing in mixtures of pesticides have not been well elucidated. Strobilurin and succinate dehydrogenase inhibitor (SDHI) fungicides are new types of commonly used pesticides. In this study, effects of three strobilurins (pyraclostrobin, trifloxystrobin and azoxystrobin), two SDHIs (isopyrazam and bixafen), and their mixtures on X. tropicalis embryos were fully investigated. Results showed that exposure to individual fungicides induced lethal and teratogenetic effects; and malformed embryos displayed similar phenotypes including microcephaly, hypopigmentation, somite segmentation and narrow fin. Exposure to two strobilurins or two SDHIs at equitoxic concentrations caused additive or synergetic effects at environmentally relevant concentrations. TU for mixtures of isopyrazam and bixafen was 0.53 and 0.30 for lethal and teratogenic toxicity, respectively. Finally, binary mixtures of strobilurins and SDHIs also exhibited additive or synergetic effects on amphibian embryos. Overall, these results reveal that the mixtures of multiple fungicides caused a higher incidence of lethality and teratogenicity of amphibian embryos, compared to a single fungicide at the corresponding doses. Our findings provide important data about the ecotoxicology of agricultural fungicides on non-target organisms, which is useful for guiding management practices for pesticides.

Comparison of phenotypic and global gene expression changes in Xenopus tropicalis embryos induced by agonists of RAR and RXR.

Retinoic acid functions through two classes of receptors, i.e., the retinoic acid receptor (RAR) and the retinoid X receptor (RXR). The difference in the role between RAR and RXR, however, are not well clarified. In the present study, we comparatively investigated the phenotypic and global gene expression changes in Xenopus tropicalis embryos induced by three different agonists, including a RAR selective ligand (all-trans retinoic acid, at-RA), a RXR selective ligand (fluorobexarotene, FBA) and their common ligand (9-cis retinoic acid, 9c-RA). All three agonists induced striking and similar malformations in X. tropicalis embryos at the concentrations of 5-50µg/L. Especially, the development of anterior structures and caudal region was dramatically altered. The hierarchical clustering analysis of phenotypes and gene profiles suggested that effects induced by 9c-RA separated from those by at-RA and FBA. The up-regulated genes were involved in multiple pathways while down-regulated genes were mainly related to phototransduction and tyrosine metabolism. at-RA primarily affected the retinol, glycolysis, starch and sucrose metabolisms while FBA led to disturbances in more wide-ranging pathways such as the PPAR, adipocytokine, insulin, FoxO signaling pathways, alanine, aspartate and glutamate metabolism. RXR is a heterodimeric partner for several other nuclear receptors, which opens the possibility that additional retinoid effects could be mediated by FBA, such as RXR-PPAR. Our data indicates that not only RXR-RAR but also RXR-PPAR plays important roles in the control of metabolism with retinoid treatment in X. tropicalis embryos.

The unexpected teratogenicity of RXR antagonist UVI3003 via activation of PPARγ in Xenopus tropicalis.

The RXR agonist (triphenyltin, TPT) and the RXR antagonist (UVI3003) both show teratogenicity and, unexpectedly, induce similar malformations in Xenopus tropicalis embryos. In the present study, we exposed X. tropicalis embryos to UVI3003 in seven specific developmental windows and identified changes in gene expression. We further measured the ability of UVI3003 to activate Xenopus RXRα (xRXRα) and PPARγ (xPPARγ) in vitro and in vivo. We found that UVI3003 activated xPPARγ either in Cos7 cells (in vitro) or Xenopus embryos (in vivo). UVI3003 did not significantly activate human or mouse PPARγ in vitro; therefore, the activation of Xenopus PPARγ by UVI3003 is novel. The ability of UVI3003 to activate xPPARγ explains why UVI3003 and TPT yield similar phenotypes in Xenopus embryos. Our results indicate that activating PPARγ leads to teratogenic effects in Xenopus embryos. More generally, we infer that chemicals known to specifically modulate mammalian nuclear hormone receptors cannot be assumed to have the same activity in non-mammalian species, such as Xenopus. Rather they must be tested for activity and specificity on receptors of the species in question to avoid making inappropriate conclusions.

An assay to determine the sensitive window of embryos to chemical exposure using Xenopus tropicalis.

The frog embryo teratogenesis assay-Xenopus (FETAX) is an established method to evaluate the developmental toxicity of chemicals. In FETAX, a 48 h continuous exposure is usually conducted when the X. tropicalis embryo is used as the test model. In the present study, we exposed X. tropicalis embryos to nine known teratogens for four separate 12-h periods. The embryos showed great variations in response to nine tested compounds during different exposure periods. Based on the value of the score of malformations, the most sensitive 12 h exposure periods of embryos were significantly distinguished for all the compounds with the exception of NiCl2 . The embryos were the most sensitive to retinols (e.g. all-trans-retinoic acid and 9-cis-retinoic acid) during 0-12 h and to metal compounds (e.g. triphenlytin and CdCl2) during a 24 to 36 h exposure period. In the further 3 h exposure experiment, the most sensitive period could only be determined for one of three tested compounds. Based on the present results, we proposed an assay to determine a 12 h sensitive window of embryos to chemical exposure using Xenopus tropicalis.

Microplastic Pollution in Table Salts from China.

Microplastics have been found in seas all over the world. We hypothesize that sea salts might contain microplastics, because they are directly supplied by seawater. To test our hypothesis, we collected 15 brands of sea salts, lake salts, and rock/well salts from supermarkets throughout China. The microplastics content was 550-681 particles/kg in sea salts, 43-364 particles/kg in lake salts, and 7-204 particles/kg in rock/well salts. In sea salts, fragments and fibers were the prevalent types of particles compared with pellets and sheets. Microplastics measuring less than 200 µm represented the majority of the particles, accounting for 55% of the total microplastics, and the most common microplastics were polyethylene terephthalate, followed by polyethylene and cellophane in sea salts. The abundance of microplastics in sea salts was significantly higher than that in lake salts and rock/well salts. This result indicates that sea products, such as sea salts, are contaminated by microplastics. To the best of our knowledge, this is the first report on microplastic pollution in abiotic sea products.