Understanding boron toxicity in aquatic plants (Salvinia natans and Lemna minor) in the presence and absence of EDTA.

Even though boron is a widely used element in various industries and a contributor to water pollution worldwide, few studies have examined the toxicity of boron in aquatic plants. EDTA is used to maintain aquatic plants cultures, however it is possible to modify the toxicity of metals. The objective of this study is to assess the toxicity of boron in aquatic plants and explore the impact of EDTA presence on the resulting toxic responses. Floating watermoss Salvinia natans and duckweed Lemna minor were exposed to concentrations ranging from 5 to 100 mg/L for 7 days and 1 to 60 mg/L for 3 days, respectively. Growth and photosynthetic activity parameters were investigated in the presence and absence of EDTA. Growth inhibitions in both aquatic plants were observed in a concentration-dependent manner, irrespective of the presence or absence of EDTA. For instance, based on the specific growth rate (leaves coverage), EC10 values for S. natans were calculated as 12.7 (9.9-15.3) mg/L and 8.0 (5.8-10.3) mg/L with and without EDTA, respectively. In the case of L. minor, EC10 values were calculated as 1.3 (0.8-1.89) mg/L and 2.0 (0.4-4.3) mg/L with EDTA without EDTA, respectively. Significant effects were also observed on the photosynthetic capacity, however there was no change in the increase of boron concentration. Generally, negligible effects of EDTA to the toxicity of boron were observed in the present study. By comparing toxicity results based on the presence and absence of EDTA, which is an essential element in the test medium, the results of this study are expected to be utilized for the ecological risk assessment of boron in aquatic ecosystems.

Evidence of soil particle-induced ecotoxicity in old abandoned mining area.

Ecotoxicity of heavy metals in soil is primarily associated with their bioaccessibility and bioavailability in the soil media. However, in some exceptional cases, soil ecotoxicity has been observed despite high total metal concentrations and low extractable metal concentrations in contaminated field sites; therefore, other exposure pathways must be considered. Therefore, the aim of this study was to evaluate the soil-particle induced ecotoxicity in an old mining area. We hypothesized that heavy metals, strongly adsorbed onto soil particles of consumable size for soil organisms, exhibit ecotoxicity, especially on soil particles ∼1 µm to 300 µm in size. A plant seedling assay, in vivo cytotoxicity assay using earthworm immune cells, and a metal bioconcentration assessment were performed. The results of soil particle toxicity revealed that the soil from the study area (A1-A4) had a low contribution to the soil ecotoxicity of extractable metals. For instance, the concentration of extractable arsenic was only 1.9 mg/kg soil, despite the total arsenic concentration reaching 36,982 mg/kg soil at the A1 site. The qualitative and quantitative analyses using SEM-EDX and ICP-OES, as well as principal component analyses, supported the hypothesis of the present study. Overall, the study results emphasize the importance of soil particle-induced ecotoxicity in long-term contaminated field soils. Our study results can inform on effective site-specific soil ecological risk assessment as they suggest the inclusion of soil particle-induced ecotoxicity as an important criterion in old, contaminated field sites, even when the extractable metal fraction in the field soil is low. ENVIRONMENTAL IMPLICATION: Bioaccessibility and bioavailability are primary factors contributing to the soil ecotoxicity of heavy metals. However, in some cases, such as long-term contaminated field sites, soil ecotoxicity has been confirmed even when low extractable metal concentrations were detected alongside high total metal concentrations. The findings of this study reveal that soil particles of edible size could be sources of soil ecotoxicity in the case of long-term contaminated fields with low extractable metal concentrations. The results of this study would contribute to the area of site-specific soil ecological risk assessment.

Microplastic ingestion in aquatic and soil biota: A comprehensive review of laboratory studies on edible size and intake pattern.

Microplastic contamination is ubiquitous and can be transferred through the food chain to humans. However, studies on microplastic size have mainly focused on large animals with a body length >20 mm. To address this gap, we conducted a comprehensive review of 169 laboratory studies to determine the edible size of microplastics for macrofauna and flora in aquatic and soil biota. Our findings indicate that microplastics with a size of <300 µm and 1 µm, respectively, are edible for these organisms, which are positioned at the base of the food chain. We also analyzed intake and depuration patterns and identified factors affecting microplastic ingestion. Our study fills an important knowledge gap by identifying the range of microplastic sizes that can enter the food chain and be transferred to humans. The study findings have strong implications for the ecological risk assessment of microplastics and suggest a starting point for mitigating this threat.

Microplastics promote the accumulation of negative fungal groups and cause multigenerational effects in springtails.

The environmental persistence of microplastics (MPs) is ubiquitous and problematic. Despite an increase in research on the soil ecotoxicity of MPs, the response of springtails to MP pollution remains unexplored. We hypothesized that MPs promote the accumulation of negative soil fungal groups and cause multigenerational effects in springtails. We performed a multigenerational study of high-density polyethylene MPs using springtail Folsomia candida and analyzed the soil fungal community. We found that soil entomopathogenic fungi and negative soil fungal groups accumulated in springtail F. candida due to soil MP pollution; subsequently, MPs negatively affected F. candida in the F2 generation. To the best of our knowledge, this is the first study to investigate the correlations between MP pollution, soil fungi, and fungi-feeding springtails. The study provides evidence of the accumulation of soil entomopathogenic fungi and negative soil fungal groups in F. candida caused by soil MP pollution.

Assessing the risks of silver nanoparticle-concentrated matrix application in agricultural soil: Implications for plant and soil enzymes.

The practice of reusing biosolids may be an effective strategy for addressing nutrient reuse and soil shortages. However, the accumulation of silver nanoparticles (AgNPs) in biosolids poses a major ecological hazard to organisms. In this study, we evaluated the effects of AgNPs on plant and soil enzymes in terrestrial ecosystems by exposing soybean Glycine max, as well as soil, to varying concentrations of AgNPs embedded either on the surface soil layer or throughout the soil matrix. We found that exposing the plants to 20 mg AgNPs/kg soil that was embedded throughout the soil matrix had a significant impact. Conversely, soil enzymes (dehydrogenase, urease, and fluorescein diacetate hydrolase) were affected by exposure to 20 and 80 mg AgNPs/kg of soil embedded on the surface soil layer or throughout the soil matrix at 20 mg AgNPs/kg soil. The toxic effects of AgNPs were induced by different receptor habitats. The presence of AgNPs in soil led to developmental retardation, inhibited root growth, reduced photosynthetic activity, and decreased transpiration rates in leaves. These plants also produced fewer fruits compared to soybean plants grown in regular soil without AgNPs. Moreover, most of the AgNPs soil groups demonstrated a decrease in soil enzyme activity in the surface soil; therefore, soil-capping patterns involving embedded stressors should be considered. These findings provide valuable insights that will contribute substantially to advancements in the field of biosolids for land applications.

Multigenerational effects of microplastic fragments derived from polyethylene terephthalate bottles on duckweed Lemna minor: Size-dependent effects of microplastics on photosynthesis.

The 2019 global coronavirus disease pandemic has led to an increase in the demand for polyethylene terephthalate (PET) packaging. Although PET is one of the most recycled plastics, it is likely to enter the aquatic ecosystem. To date, the chronic effects of PET microplastics (MPs) on aquatic plants have not been fully understood. Therefore, this study aimed to investigate the adverse effects of PET MP fragments derived from PET bottles on the aquatic duckweed plant Lemna minor through a multigenerational study. We conducted acute (3-day exposure) and multigenerational (10 generations from P0 to F9) tests using different-sized PET fragments (PET0-200, < 200 µm; PET200-300, 200-300 µm; and PET300-500, 300-500 µm). Different parameters, including frond number, growth rate based on the frond area, total root length, longest root length, and photosynthesis, were evaluated. The acute test revealed that photosynthesis in L. minor was negatively affected by exposure to small-sized PET fragments (PET0-200). In contrast, the results of the multigenerational test revealed that large-sized PET fragments (PET300-500) showed substantial negative effects on both the growth and photosynthetic activity of L. minor. Continuous exposure to PET MPs for 10 generations caused disturbances in chloroplast distribution and inhibition of plant photosynthetic activity and growth. The findings of this study may serve as a basis for future research on the generational effects of MPs from various PET products.

Salvinia natans microplate assay: A simple and efficient method for evaluating aquatic toxicity.

This study proposes a microplate assay for chemical toxicity evaluation using S. natans; this assay is a miniaturization of a culture dish assay from previous studies. The optimal initial frond number of S. natans in the microplate assay was established and the feasibility of the microplate assay was evaluated. To evaluate the applicability of the developed method, S. natans was exposed to six model chemicals (one metalloid and five heavy metals) and the specific growth rate and photosynthetic activity were compared. We determined that the optimal initial frond number was four fronds for the microplate assay. Additionally, toxicity sensitivity to six model chemicals revealed similar results to the culture dish assay. Therefore, we propose that the S. natans microplate assay is a simple and effective method to assess ecotoxicity as it requires less test solution and area than the culture dish assay. This methodology can be expandable to ecotoxicological research.

Acute and multigenerational effects of petroleum- and cellulose-based microfibers on growth and photosynthetic capacity of Lemna minor.

Comparative toxicological assessment studies on the effects of petroleum- and cellulose-based microfibers on aquatic plants are limited. Therefore, we evaluated the acute and 10-generational toxicological effects of two types of petroleum- and cellulose-based microfibers on the duckweed Lemna minor. Plant growth and photosynthesis parameters were monitored as bioindicators. The multigenerational test revealed the following ranking of the microfibers according to the severity of their effects on L. minor: polypropylene > lyocell > viscose > polyethylene terephthalate. The acute tests revealed a significant increase in the energy required to initiate photosynthesis, although the growth of L. minor was not adversely affected by any microfiber. Both petroleum- and cellulose-based microfibers induced adverse effects on the growth and photosynthesis of L. minor in multigenerational tests. The results of the generational tests contribute to the understanding of the long-term adverse effects of microfibers on aquatic plants.

Earthworm half-pipe assay: A new alternative in vivo skin corrosion test using invertebrates.

As a result of the efforts to introduce the principle of the 3Rs (replacement, reduction, and refinement) into animal testing, alternative in vitro skin corrosion test methods have been developed and standardized globally. However, alternative in vitro skin corrosion test methods have some limitations in terms of the use of humanely killed rats or commercial models and kits. The present study focused on the applicability of invertebrates as alternative in vivo skin models. Even though earthworm skin comprises the same biomolecules as human skin, the possibility of using earthworm skin as an alternative for skin testing remains unexplored. In this study, we developed a half-pipe tool for earthworm skin corrosion testing and optimized the test protocol. Subsequently, the applicability of the earthworm half-pipe assay for corrosion testing with six chemicals, including inorganic acids, organic acids, and alkalis, was investigated using stereomicroscopy and electron microscopy. It was observed that the specific concentrations for earthworm skin corrosion were lower than those for animal or in vitro tests. Therefore, the sensitivity of the earthworm half-pipe assay indicates that it could be useful as a screening tool before conducting in vivo animal tests or in vitro skin tests. This new method can contribute to research on alternative skin corrosion tests by reducing ethical issues, time, and cost while achieving effective results.

Length- and polymer-dependent ecotoxicities of microfibers to the earthworm Eisenia andrei.

Microfibers are widespread environmental pollutants introduced to the soil environment because of the increasing use of plastic polymers. However, research on the soil ecotoxicity of microfibers is limited, especially when compared to research on their aquatic toxicity. This study investigated the effects of sub-chronic microfiber exposure on the earthworm Eisenia andrei. We compared the effects of three types of microfibers: short lyocell microfibers (231 ± 126 µm long), short polypropylene microfibers (191 ± 107 µm long), and long polypropylene microfibers (891 ± 141 µm long). After exposure for 21 days, earthworm survival, coelomocyte viability, cast microbial viability, and gut microbial viability were assessed, and a histopathological examination of the digestive tract and reproductive tissues was conducted. In addition, long polypropylene microfibers egested by the earthworms were collected to explore the possibility of earthworm-driven biofragmentation. Results indicated that high exposure concentration (1000 mg/kg dry soil) negatively affected earthworm coelomocytes and intestinal tissue, gut, and cast microbiomes. Although all three microfiber types reduced earthworm survival, the short polypropylene microfibers were more toxic to the earthworms than the long polypropylene microfibers or short lyocell microfibers, which indicated that size-dependent soil ecotoxicity was induced. PP microfibers were found to more negatively affect cast microbial activity and intestinal tissue than lyocell microfibers, indicating polymer-dependent soil ecotoxicity potential against earthworm species. This study provides evidence that synthesized microfibers cause cytotoxicity and decrease gut microbiome viability in earthworms, and that they can be biofragmented by earthworms.

Site-specific ecological risk assessment of metal-contaminated soils based on the TRIAD approach.

Ecological risk assessment based on scientific data is crucial for understanding causal relationships between chemical pollution and environmental risks. Simultaneously, a balance is required between socioeconomic factors and scientific evidence. The TRIAD approach, which incorporates three lines of evidence (LoE)-chemical (Chem-LoE), ecotoxicological (Ecotox-LoE), and ecological (Eco-LoE)-was applied in five sites of an abandoned mine for site-specific soil ecological risk assessment (SERA). In combination, the three LoEs showed that two sites had extremely high risks, one site had moderate risk, and the other site had low risk. At all sites, Chem-LoE exhibited high-integrated risk values. In Ecotox-LoE and Eco-LoE, some species were not affected despite high metal concentrations in the soil samples collected from the sites, indicating that the bioavailability of metals differed according to the physiochemical properties of the soil medium. This study is significant as multiple analyses were performed considering ecosystem structure to reduce uncertainty in SERA. The results provide information to support effective decision-making risk management to protect the soil ecosystem. Moreover, these findings will be useful in establishing policies and priorities for soil risk management.

Sub-acute exposure to nanoplastics via two-chain trophic transfer: From brine shrimp Artemia franciscana to small yellow croaker Larimichthys polyactis.

This study investigated the trophic transfer of nanoplastics in marine food chains. We fed nanoplastic-exposed Artemia franciscana (brine shrimp) to Larimichthys polyactis (small yellow croaker) daily for eight days. Subsequently, the overall health condition, histopathological damage to the liver and digestive tract, and swimming ability of the fish were measured. After the sub-acute exposure to nanoplastics via trophic transfer, the fish showed inhibited growth, severe liver damage, as well as a poorer swimming ability compared to the control. The swimming ability was especially affected, in terms of the overall movement as well as thigmotaxis. The results thus clarified that even an indirect exposure to nanoplastics could induce neurotoxic effects and affect the swimming ability of the fish. As fish are well-known human food resources, the possibility of such trophic transfers affecting higher trophic level organisms, such as humans, cannot be ruled out.

Species sensitivity distributions for ethylparaben to derive protective concentrations for soil ecosystems.

Ethylparaben is used as an antifungal preservative. Although some countries have implemented regulations for human exposure to parabens, environmental regulations for ethylparaben have not been established. This study provides new toxicological data for ethylparaben, for which data regarding soil organisms were previously lacking. Although ethylparaben toxicity has been reported in other species, we present herein the first comprehensive study of its toxicity in soil organisms. We used 12 test species (Lycopersicon esculentum, Vigna radiata, Hordeum vulgare, Oryza sativa, Eisenia andrei, Folsomia candida, Lobella sokamensis, Caenorhabiditis elegans, Chlamydomonas reinhardtii, Chlorococcum infusionum, Chlorella sorokiniana, Chlorella vulgaris) from eight taxonomic groups for acute bioassays and nine test species (L. esculentum, V. radiata, H. vulgare, O. sativa, C. reinhardtii, C. infusionum, C. sorokiniana, and C. vulgaris) from five taxonomic groups for chronic bioassays. A suite of acute and chronic toxicity tests, using 21 soil species, was conducted to estimate EC50 values, which facilitated the construction of species sensitivity distributions (SSDs) and the calculation of protective concentrations (PCs). Acute and chronic PC95 values (protective concentration for 95% of species) for ethylparaben were estimated to be 14 and 5 mg/kg dry soil, respectively. To the best of our knowledge, this is the first study to evaluate the toxicity of ethylparaben to soil species and derive PCs for soil ecosystems based on SSDs. Therefore, the data presented in this study can be used as a basis for further investigations of paraben toxicity to the soil environment.

Critical review of environmental impacts of microfibers in different environmental matrices.

Microfibers, as globally emerging environmental pollutants, have received increasing attention over recent years. In ecosystems, natural textile microfibers are the predominant fibers. The inadvertent ingestion of microplastics by organisms in an ecosystem provides a channel for microfibers to enter biological webs. Based on existing research on microfibers, this review summarizes the potential adverse impacts of microfibers on organisms living in marine, freshwater, and soil ecosystems, and provides a brief introduction to the source of microfibers, as well as the related current status and future challenges. Although previous studies have recorded the adverse effects of microfibers on ecosystems, there remains a lack of evidence on the toxic effects of microfibers on the primary producer level in food chains (e.g., phytoplankton). This is essential, as the long-term effects of microfiber toxicity on different ecosystems ultimately affect human health. The analysis of microfiber toxicity paves the way for the field of environmental research and provides future perspectives for researchers in the fields of ecotoxicology and microplastics.

Post COVID-19 pandemic: Biofragmentation and soil ecotoxicological effects of microplastics derived from face masks.

Because of the COVID-19 pandemic, used face masks have increasingly littered the environment and are causes for concern since they are commonly made of plastics such as polypropylene. Understanding production of microplastics from face masks is essential for predicting the post COVID-19 pandemic impact on the soil ecosystem. We investigated the generation of nanofibers from meltblown face mask filters (MB filters) and their adverse effects on soil species, particularly the earthworm and springtail. Results of MB filter soil bioassays at a high concentration (1000 mg/kg dry soil) suggest inhibited reproduction and stunted growth in springtails, decreased intracellular esterase activity in earthworm coelomocytes, and inhibited spermatogenesis in male earthworm reproductive tissues. Moreover, it was estimated that generation of nanofibers from microfibers and fragments of MB filters might occur in the soil ecosystem post COVID-19. This study does not oppose the use of face masks but aims to encourage appropriate disposal of the masks. Preservation of human health and the ecosystem should be prioritized even amidst the COVID-19 pandemic.

Combined exposure to microplastics and zinc produces sex-specific responses in the water flea Daphnia magna.

Microplastics are ubiquitous environmental pollutants and a great threat to the aquatic environment. Due to their small size (ranging from 1 µm to 5 mm), microplastics be easily ingested by a wide range of organisms and can serve as a vector for various contaminants. In this study, additive or possible synergistic effects of microplastics and zinc were demonstrated through sex-specific alterations in behavior, redox status, and modulation of detoxification-related genes in Daphnia magna, with males being more sensitive than females with stronger modulations of antioxidant responses, particularly on glutathione S-transferases expressions. Furthermore, we demonstrated microplastics may act as vectors for metals (Zn2+) in the aquatic environment in D. magna, with reduced bio-concentration of the total Zn concentration, inducing greater toxicity. Our findings demonstrated synergistic toxicity of the heavy metal Zn and microplastics and could contribute to greater understanding of sex-specific effects of microplastics in aquatic organisms.

Comparative toxicity of potential leachates from perovskite and silicon solar cells in aquatic ecosystems.

Globally, perovskite solar cells (PSCs) represent a third-generation photovoltaic technology that is being increasingly implemented and commercialized. However, the biological impacts of leachates from PSCs are poorly understood. Therefore, the aim of this study was to investigate the ecotoxicity of PSC leachates compared with that of commercial Si-based solar cell (SBSC) leachates. We performed leaching assessments and aquatic bioassays using internationally recommended test species and measured and compared the ecotoxicity of PSC and SBSC leachates. As a result of the leaching analyses, Si, Pb, and Al were found to be the most leached elements from broken PSCs and SBSCs. The bioassays indicated that polycrystalline SBSC (p-Si) and monocrystalline SBSC (m-Si) leachates were more toxic to fish embryos than the PSC leachates and that water fleas were sensitive to m-Si leachates, but less sensitive to PSC and p-Si leachates. In addition, principle component analyses indicated that the ecotoxicity of solar cell leachates was related to either the Pb or Si content. This is the first comparative study of the potential ecotoxicity of PSC and SBSC leachates in aquatic ecosystems, and the results of which can be used in the environmentally safe commercialization of solar cells.

Assessing potential indicator of endocrine-disrupting property of chemicals using soil invertebrates.

Vitellogenin has been regarded as an acceptable indicator for evaluating the endocrine-disrupting property of chemicals using fish. However, the endocrine-disrupting property of chemicals has been rarely evaluated using soil species. This study aimed to find evidence that endocrine-disrupting chemicals (including the natural hormones estradiol and dihydrotestosterone) can affect the reproductive organs of earthworms. Earthworms were exposed to 17ß-estradiol, dihydrotestosterone, bisphenol A, and methylparaben for seven days. The four EDCs inhibited normal oogenesis and maturation of oocytes in earthworm ovary, and dihydrotestosterone and bisphenol A were observed to damage earthworm seminal vesicle tissues and inhibit normal spermatogenesis. The evidence showed that the tested EDCs have an adverse effect on female and male reproductive systems of soil invertebrates. The results suggest that the evaluations of oogenesis and spermatogenesis in the ovary and seminal vesicles of earthworms are useful indicators for investigating the endocrine-disrupting property of chemicals. Additionally, our results encourage further studies on developing novel indicators using soil invertebrates to evaluate the effects of the toxicity of endocrine-disrupting chemicals on the soil ecosystem.

Sublethal toxicity of PbI2 in perovskite solar cells to fish embryos (Danio rerio and Oryzias latipes): Deformity and growth inhibition.

Pb-based perovskite in solar cells is a source of PbI2. The objective of this study was to characterize the embryonic toxicity of PbI2, a potentially leachable chemical and hazardous material, for two fish species (zebrafish and Japanese medaka). A series of measurements were performed to assess mortality, abnormalities (deformities and other pathological changes), hatchability, and growth inhibition. The results obtained showed that the toxicities observed were predominantly associated with Pb2+ and I-. Therefore, given the potential ecotoxicity of PbI2, precautions should be taken to prevent its release during the breakage and disposal of Pb-based perovskite solar cells.

Multigenerational effects of polyethylene terephthalate microfibers in Caenorhabditis elegans.

Microfibers (MFs) have recently become an increasingly prevalent pollutant in ecosystems and pose a direct threat to organisms and an indirect threat via adsorption of other pollutants. Here, we used Caenorhabditis elegans to study multigenerational effects of polyethylene terephthalate (PET) MFs (diameter 17.4 µm) by observing the maternal generation (P0) to the seventh offspring generation (F7) with continuous MF exposure. Exposure to 250-µm PET MFs decreased locomotion behavior and induced intestinal reactive oxygen species (ROS) in the P0 generation compared with other PET MF sizes. Moreover, no notably negative effects on survival were observed in any generation during continuous exposure to 250-µm PET MFs. However, the reproduction rate clearly decreased in the F2 and F3 generations but gradually recovered in the F4-F7 generations. Developmental abnormalities showed a close relationship with body length. Although some recovery was confirmed, there were significant decreases in body length in the F2-F5 generations. Interestingly, growth inhibition was also observed in the F6 generation without MF exposure. ROS production and dermal damage in the P0-F5 generations might have resulted in the toxicological responses. To the best of our knowledge, this is the first study to provide evidence of multigenerational toxicity of MFs in C. elegans.