The soil plastisphere.

Understanding the effects of plastic pollution in terrestrial ecosystems is a priority in environmental research. A central aspect of this suite of pollutants is that it entails particles, in addition to chemical compounds, and this makes plastic quite different from the vast majority of chemical environmental pollutants. Particles can be habitats for microbial communities, and plastics can be a source of chemical compounds that are released into the surrounding environment. In the aquatic literature, the term 'plastisphere' has been coined to refer to the microbial community colonizing plastic debris; here, we use a definition that also includes the immediate soil environment of these particles to align the definition with other concepts in soil microbiology. First, we highlight major differences in the plastisphere between aquatic and soil ecosystems, then we review what is currently known about the soil plastisphere, including the members of the microbial community that are enriched, and the possible mechanisms underpinning this selection. Then, we focus on outlining future prospects for research on the soil plastisphere.

Toxicity of Aged Paint Particles to Soil Ecosystems: Insights from Caenorhabditis elegans.

Despite the extensive global consumption of architectural paint, the toxicological effects of aged exterior paint particles on terrestrial biota remain largely uncharacterized. Herein, we assessed the toxic effect of aged paint particles on soil environments using the nematode Caenorhabditis elegans (C. elegans) as a test organism. Various types of paint particles were generated by fragmentation and sequential sieving (500-1000, 250-500, 100-250, 50-100, 20-50 µm) of paint coatings collected from two old residential areas. The paint particles exerted different levels of toxicity, as indicated by a reduction in the number of C. elegans offspring, depending on their size, color, and layer structure. These physical characteristics were found to be closely associated with the chemical heterogeneity of additives present in the paint particles. Since the paint particle sizes were larger than what C. elegans typically consume, we attributed the toxicity to leachable additives present in the paint particles. To assess the toxicity of these leachable additives, we performed sequential washings of the paint particles with distilled water and ethanol. Ethanol washing of the paint particles significantly reduced the soil toxicity of the hydrophobic additives, indicating their potential environmental risk. Liquid chromatography-mass spectrometry analysis of the ethanol leachate revealed the presence of alkyl amines, which exhibited a high correlation with the toxicity of the paint particles. Further toxicity testing using an alkyl amine standard demonstrated that a paint particle concentration of 1.2% in soil could significantly reduce the number of C. elegans offspring. Our findings provide insights into the potential hazards posed by aged paint particles and their leachable additives in the terrestrial environment.

Soil Storage Conditions Alter the Effects of Tire Wear Particles on Microbial Activities in Laboratory Tests.

In this study, we focused on the fact that soil storage conditions in the laboratory have never been considered as a key factor potentially leading to high variation when measuring effects of microplastics on soil microbial activity. We stored field-collected soils under four different conditions [room-temperature storage, low-temperature storage (LS), air drying (AD), and heat drying] prior to the experiment. Each soil was treated with tire wear particles (TWPs), and soil microbial activities and water aggregate stability were investigated after soil incubation. As a result, microbial activities, including soil respiration and three enzyme activities (ß-glucosidase, N-acetyl-ß-glucosaminidase, and phosphatase), were shown to depend on soil storage conditions. Soil respiration rates increased with the addition of TWPs, and the differences from the control group (no TWPs added) were more pronounced in the AD TWP treatment than in soils stored under other conditions. In contrast, phosphatase activity followed an opposing trend after the addition of TWPs. The AD soil had higher phosphatase activity after the addition of TWPs, while the LS soil had a lower level than the control group. We suggest that microplastic effects in laboratory experiments can strongly depend on soil storage conditions.

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.

Microplastics disrupt accurate soil organic carbon measurement based on chemical oxidation method.

Microplastics are widespread contaminants in soils and terrestrial ecosystems in many areas worldwide. In this study, we measured soil organic carbon (SOC) and soil organic matter (SOM) in microplastic-treated soils to determine if the presence of microplastics could affect the accuracy of carbon-based soil quality indicator measurements. Six different sizes and types of microplastics were selected, and six soil samples were used to evaluate the impacts. Treating soil with polyethylene and low-density polyethylene significantly increased SOC (p < 0.05) when measured with the modified Walkley & Black method; microplastic addition (0.01%, v/v) increased SOC by >40% compared to control organic carbon-poor soil (<10.0 g kg-1). We conclude that the microplastics can disrupt the accurate measurement of SOC. Likely, the physicochemical treatment used in the SOC measurement process can cause the organic compounds and/or carbon complexes to be extracted from microplastics, and this can affect the results. Considering that SOC is a main indicator for assessing soil quality and the global carbon cycle, overestimations caused by microplastic contamination should be further discussed to identify appropriate ways to deal with microplastics as a new carbon source in the environment.

Selection of Ovalbumin-specific Binding Peptides through Instant Translation in Ribosome Display Using E. coli Extract.

In vitro selection has been widely used to generate molecular-recognition elements in analytical sciences. Although reconstituted types of in vitro transcription and translation (IVTT) system, such as PURE system, are nowadays widely used for ribosome display and mRNA/cDNA display, use of E. coli extract is often avoided, presumably because it contains unfavorable contaminants, such as ribonuclease. Nevertheless, the initial speed of protein translation in E. coli extract is markedly faster than that of PURE system. We thus hypothesized that E. coli extract is more appropriate for instant translation in ribosome display than PURE system. Here, we first revisit the potency of E. coli extract for ribosome display by shortening the translation time, and then applied the optimized condition for selecting peptide aptamers for ovalbumin (OVA). The OVA-binding peptides selected using E. coli extract exhibited specific binding to OVA, even in the presence of 50% serum. We conclude that instant translation in ribosome display using E. coli extract has the potential to generate easy-to-use and economical molecular-recognition elements in analytical sciences.

Determination of hazardous concentrations of 2,4-dinitrophenol in freshwater ecosystems based on species sensitivity distributions.

2,4-dinitrophenol (2,4-DNP) is a phenolic compound used as a wood preservative or pesticide. The chemical is hazardous to freshwater organisms. Although 2,4-DNP poses ecological risks, only a few of its aquatic environmental risks have been investigated and very limited guidelines for freshwater aquatic ecosystems have been established by governments. This study addresses the paucity of 2,4-DNP toxicity data for freshwater ecosystems and the current lack of highly reliable trigger values for this highly toxic compound. We conducted acute bioassays using 12 species from nine taxonomic groups and chronic assays using five species from four taxonomic groups to improve the quality of the dataset and enable the estimation of protective concentrations based on species sensitivity distributions. The acute and hazardous concentrations of 2,4-DNP in 5% of freshwater aquatic species (HC5) were determined to be 0.91 (0.32-2.65) mg/L and 0.22 (0.11-0.42) mg/L, respectively. To the best of our knowledge, this is the first report of a suggested chronic HC5 for 2,4-DNP and it provides the much-needed fundamental data for the risk assessment and management of freshwater ecosystems.

Effects of Different Microplastics on Nematodes in the Soil Environment: Tracking the Extractable Additives Using an Ecotoxicological Approach.

With increasing interest in the effects of microplastics on the soil environment, there is a need to thoroughly evaluate the potential adverse effects of these particles as a function of their characteristics (size, shape, and composition). In addition, extractable chemical additives from microplastics have been identified as an important toxicity pathway in the aquatic environment. However, currently, little is known about the effects of such additives on the soil environment. In this study on nematodes (Caenorhabditis elegans), we adopted an ecotoxicological approach to assess the potential effects of 13 different microplastics (0.001-1% of soil dry weight) with different characteristics and extractable additives. We found that poly(ethylene terephthalate) (PET) fragments and polyacrylicnitrile (PAN) fibers show the highest toxicity, while high-density polyethylene (HDPE), polypropylene (PP), and polystyrene (PS) fragments induced relatively less adverse effects on nematodes. In addition, low-density polyethylene (LDPE) induced no toxicity within our test concentration range for the acute period. Acute toxicity was mainly attributed to the extractable additives: when the additives were extracted, the toxic effects of each microplastic disappeared in the acute soil toxicity test. The harmful effects of the LDPE films and PAN fibers increased when the microplastics were maintained in the soil for a long-term period with frequent wet-dry cycles. We here provide clear evidence that microplastic toxicity in the soil is highly related to extractable additives. Our results suggest that future experiments consider extractable additives as key explanatory variables.

Edible size of polyethylene microplastics and their effects on springtail behavior.

Many reliable studies have provided evidence of microplastic ingestion by soil organisms. However, further research is required to determine the edible size of microplastics, especially given the ubiquity of microplastics and their adverse effects on the soil environment. Determining the size range of microplastics that can be ingested by soil organisms is crucial for the prediction of the exposure route and toxicity mechanisms of microplastics in soil. Springtails, organisms prevalent in a wide variety of soil ecosystems, can ingest or transport microplastics; however, direct evidence for this has not been reported. To address this knowledge gap, we designed dietary exposure experiments under laboratory conditions, using the springtail species Folsomia candida. The springtails were administered polyethylene microplastics in three different sizes (2, 34, and 66 µm) via their food for a short period of time; we further observed the intestinal presence of microplastics via fluorescence microscopy to determine the maximum edible size. We evaluated the effects of ingested microplastics on springtails by quantifying their moving behavior. The results show that the edible size of microplastics is < 66.0 ± 10.9 µm, and microplastics smaller than this can significantly reduce the velocity and distance of springtail movement by 74% ± 38% compared with the control group. Based on this finding, the broader fate and toxicity of microplastics in soil environments can be estimated. Furthermore, the average velocity and distance of springtail movement decreases in response to microplastic ingestion, highlighting the negative effects of microplastics on soil organisms.

Multispecies bioassay of propylparaben to derive protective concentrations for soil ecosystems using a species sensitivity distribution approach.

Propylparaben is widely used as a preservative in pharmaceuticals and personal care products and is ultimately excreted by the human body. Thus, propylparaben reaches sewage and enters the soil environment by sludge fertilization and wastewater irrigation. However, there are few existing studies on the toxicity and risks of such chemicals in terrestrial environments. In this study, a multispecies bioassay for propylparaben was performed and protective concentrations (PCs) were derived based on toxicity values by probabilistic ecological risk assessment. Acute and chronic bioassays were conducted on 11 species in eight taxonomic groups (Magnoliopsida, Liliopsida, Clitellata, Entognatha, Entomobryomorpha, Chromadorea, Chlorophyceae, Trebouxiophyceae). Based on the toxicity values calculated, the PC95 values for acute and chronic SSDs were 13 and 6 mg/kg dry soil, respectively. Toxicity varied among taxa, with soil algae emerging as the most sensitive to propylparaben. This may be attributable to differences in exposure pathways among species. The exposure pathway of propylparaben can be altered by adsorption to soil particles. As parabens are presently under-regulated globally in terms of their environmental effects, our findings can serve as the basis to propose standard values for environmental protection.

Application of a soil quality assessment system using ecotoxicological indicators to evaluate contaminated and remediated soils.

The deterioration of soil quality owing to human activities results in adverse effects on the soil ecosystem. This study developed a systematic method to quantitatively evaluate soil quality based on physical, chemical, biological, and ecotoxicological indicators and proposed the soil quality assessment and management system. This system consists of step-by-step processes, including indicator classification, indicator measurement, scoring and weighting, and soil quality index (SQI) calculation. The novel strategy included the usage of authentic ecotoxicological indicators for realistically interpreting soil quality assessment results. This study used five ecotoxicological indicators, including earthworm survival, enzyme activities, nematode reproduction, plant germination and growth, soil algal biomass, and soil algal photosynthetic capacity. Relatively higher SQI values than those corresponding to the actual soil quality status would be obtained without considering the ecotoxicological indicators. We conclude that the use of ecotoxicological indicator can help in soil quality assessment even under extreme soil quality conditions, such as highly contaminated or physically and chemically remediated soils.

Size-dependent effects of polystyrene plastic particles on the nematode Caenorhabditis elegans as related to soil physicochemical properties.

Plastic polymers are widely used in various applications and are thus prevalent in the environment. Over time, these polymers are slowly degraded into nano- and micro-scale particles. In this study, the free-living nematode, Caenorhabditis elegans, was exposed to polystyrene particles of two different sizes (42 and 530 nm) in both liquid and soil media. The number of offspring significantly (p < 0.05) decreased at polystyrene concentrations of 100 mg/L and 10 mg/kg in liquid and soil media, respectively. In soil media, but not liquid media, C. elegans was more sensitive to the larger particles (530 nm) than the smaller particles (42 nm), and the median effective concentration (EC50) values of the 42 and 530 nm-sized particles were found to be > 100 and 14.23 (8.91-22.72) mg/kg, respectively. We performed the same toxicity bioassay on five different field-soil samples with different physicochemical properties and found that the size-dependent effects were intensified in clay-rich soil samples. A principal component analysis showed that the bulk density, cation exchange capacity, clay content, and sand content were the dominant factors influencing the toxicity of the 530 nm-sized polystyrene particles. Therefore, we conclude that the soil composition has a significant effect on the toxicity induced by these 530 nm-sized polystyrene particles.

Preparation of azidophenyl-low molecular chitosan derivative micro particles for enhance drug delivery.

Systems for delivering damaged tissue by immobilization of a bioactive substance or a protein drug for rapid recovery of a patient are being studied. To immobilize drugs using natural polymer, photo-immobilization method has been designed. Immobilization through photo-reaction is a new technology that stabilizes drugs or growth factors for sustained release. Introduction of photo-reactive functional groups into biocompatible natural polymers produces materials applicable to the medical field. Since chitosan is a natural polymer with stability and biocompatibility, this study attempts to use chitosan as a mediator of drug delivery. In addition, If the form of the immobilized biomaterial is made into a micro-sized particle, it can be utilized as an injectable material in addition to the stability of the photo-immobilization. In photo-immobilization in particle form, the probability of exposure to the enzyme in the body is lower than if it is injected into the body in the conventional free state. In addition, since it can be freely injected into a desired target site, it can be used for various medical applications. Therefore, it is expected that various effects of growth factors and drugs can be utilized and additional effects can be obtained by photo-immobilization together with various effects.

Changes in soil properties after remediation influence the performance and survival of soil algae and earthworm.

Previous research on soil remediation focused on soil properties and not on its effects on soil ecosystems. The present study investigated the adverse effects of soil physicochemical changes due to remediation on the biological indicators Chlorococcum infusionum and Chlamydomonas reinhardtii (algae) and Eisenia andrei (earthworm). Soil physicochemical properties, concentrations of total, bioavailable, and water-soluble heavy metals in soil were measured before and after remediation. Changes in soil pH, electrical conductivity, total nitrogen, and total phosphorous immediately after soil remediation were the primary causes of the biomass and photosynthetic activity inhibition observed in C. infusionum and C. reinhardtii, and the survival, normality, and burrowing behavior decrease observed in E. andrei in remediated soils showing dramatic changes in those properties. These findings suggest that remediated soils need some time to recover before restoring their functions, although heavy metals are no longer contaminating the soil.

Soil microplastics inhibit the movement of springtail species.

Previous studies have indicated the means by which micro-sized plastic particles may affect the soil environment, and this could be linked to the behavior of plastics in the soil system and how these particles are influenced by biological responses. Soil-dwelling organisms play a key role in modifying the soil system by constructing bio-pores, and these structural changes are potentially related to the behavior of plastic particles. In this study, we found that micro-sized plastic particles moved into bio-pores within seconds, and that this influx disrupted the movement of springtails (Lobella sokamensis). The springtails moved to avoid becoming trapped, and this behavior created bio-pores in the soil system. The influx of plastic particles into these cavities subsequently immobilized the springtails within. This phenomenon was observed at low a concentration of plastic particles (8 mg/kg), and it likely occurs in actual soil environments. The findings of this study indicate that the behavior of plastic particles in the soil not only disrupts the movement of springtails but also has wider implications for effective management of soils.

A simple and efficient method for separation of low-density polyethylene films into different micro-sized groups for laboratory investigation.

Microplastics are abundant in both aquatic and terrestrial environments. While they have received much recent attention because of their effects on ecosystems, their true impact on natural environments remains difficult to assess because of the problems associated with processing them in the laboratory. In this study, we designed and implemented a new, vacuum-based method of separating different sizes of low-density polyethylene films. Using multiple sieve sizes, we achieved consistent recovery of the desired size fractions using this method. The vacuum suction (VS) system consisted of two differently sized cylindrical sieves of 500 µm (or 200 µm) and 65 µm, allowing film samples between 65 and -500 µm and 65-200 µm to be collected. The VS systems successfully separated microplastic film samples into small area ranges of 0.015-0.065 mm2, and the film areas showed different distributions for each sample from the different VS systems. This system provided an easy, rapid, and low-labor means of processing different sizes of microplastics via an innovative method. Further research into the effects of microplastics on natural environments is critically needed, and the laboratory separation of different size fractions of microplastics facilitates such endeavors.

Zebrafish can recognize microplastics as inedible materials: Quantitative evidence of ingestion behavior.

This study investigated the ingestion behavior of zebrafish exposed to microplastic particles (MPs) at different concentrations, presented alone or in a mix with food particles. Zebrafish showed spitting behavior after ingesting micro-sized (247.5 µm) polyethylene particles (i.e., MPs), with prey-capture time increasing when food and MPs were supplied simultaneously. Fish were compelled to ingest MPs with food, and the accumulation percentage (ingested particles/supplied particles) was quantified as 0.5 to 9.4% with increasing food volume. However, the accumulation percentage was determined as 0.0 to 1.0% under exposure to MPs only, and as 3.8 to 4.3% at high MP concentrations when the food concentration was fixed. Overall, these results demonstrate that small freshwater organisms can recognize that MPs are not food items. Under laboratory conditions, zebrafish rarely discriminate between food and MPs when both are presented together, and the indiscriminate feeding behavior becomes clearer as more food is available.

Preparation of drug-immobilized anti-adhesion agent using visible light-curable alginate derivative containing furfuryl group.

This study demonstrated the anti-adhesion and wound healing effect of a visible light curable anti-adhesion agent using an alginate derivative modified with a furfuryl moiety. Visible light-curable furfuryl alginate (F-Alg) was prepared in conjugation with alginate and furfurylamine by an amide coupling reaction, and the conjugated F-Alg was characterized by 1H NMR analysis. The cytotoxicity, cell adhesion, and cell permeability of the F-Alg were evaluated for use in anti-adhesion applications. Drug immobilization and protein release were assessed to verify whether the alginate derivatives and drugs were photo-immobilized. In in vivo anti-adhesion testing, the new anti-adhesion agent prepared in this study acted as a physical protective layer by forming a biofilm on the surgical site. Additionally, along with gradual decomposition of the photo-crosslinked alginate derivative, the immobilized drug was released, and additional effects such as accelerated wound healing are expected. Thus, visible light-curable F-Alg has good application potential as an anti-adhesion agent.