PBAT is biodegradable but what about the toxicity of its biodegradation products?

CONTEXT: Poly(butylene adipate-co-terephthalate) (PBAT) is a biodegradable plastic. It was introduced to the plastics market in 1998 and since then has been widely used around the world. The main idea of this research is to perform quantum chemical calculations to study the potential toxicity of PBAT and its degradation products. We analyzed the electron transfer capacity to determine its potential toxicity. We found that biodegradable products formed with benzene rings are as good electron acceptors as PBAT and OOH•. Our results indicate that the biodegradation products are potentially as toxic as PBAT. This might explain why biodegradation products alter the photosynthetic system of plants and inhibit their growth. From this and other previous investigations, we can think that biodegradable plastics could represent a potential environmental risk. METHODS: All DFT computations were performed using the Gaussian16 at M062x/6-311 + g(2d,p) level of theory without symmetry constraints. Electro-donating (ω-) and electro-accepting (ω +) powers were used as response functions.

Bio-based microplastic polylactic acid exerts the similar toxic effects to traditional petroleum-based microplastic polystyrene in mussels.

Microplastics (MPs) have accumulated in the oceans, causing adverse effects on marine organisms and the environment. Biodegradable polylactic acid (PLA) is considered as an excellent substitute for traditional petroleum-based plastics, but it is difficult to degrade completely and easily become MPs in the marine environment. To test the ecological risk of bio-based PLA, we exposed thick-shelled mussels (Mytilus coruscus) to bio-based PLA and petroleum-based polystyrene (PS) (at 102, 104, and 106 particles/L) for 14 days. The significant increase in enzyme activities related to oxidative stress and immune response showed that mussels were under physiological stress after MP ingestion. While enzyme activities of nerve conduction and energy metabolism were significantly disturbed after exposure. Meanwhile, normal physiological activities in respiration, ingestion and assimilation were also suppressed in association with enzyme changes. The negative effects of PS and PLA in mussels were not differentiated, and further integration analysis of integrated biomarker response (IBR) and principal component analysis (PCA) also showed that PLA would induce adverse effects in mussels and ecological risks as PS, especially at environmental concentrations. Therefore, it is necessary to pay more attention to the environmental and ecological risk of bio-based MP PLA accumulating in the marine environment.

Comparative assessment of the acute toxicity of commercial bio-based polymer leachates on marine plankton.

Conventional plastics have become a major environmental concern due to their persistence and accumulation in marine ecosystems. The development of potential degradable polymers (PBP), such as polyhydroxyalkanoates (PHAs) and polylactic acid (PLA), has gained attention as an alternative to mitigate plastic pollution, since they have the potential to biodegrade under certain conditions, and their production is increasing as replacement of conventional polyolefins. This study aimed to assess and compare the toxicity of leachates of pre-compounding PBP (PLA and the PHA, polyhydroxybutyrate-covalerate (PHBv)) and polypropylene (PP) on five marine planktonic species. A battery of standard bioassays using bacteria, microalgae, sea urchin embryos, mussel embryos and copepod nauplii was conducted to assess the toxicity of leachates from those polymers. Additionally, the presence of chemical additives in the leachates was also verified through GC-MS and LC-HRMS analysis. Results showed that PHBv leachates exhibited higher toxicity compared to other polymers, with the microalgae Rhodomonas salina, being the most sensitive species to the tested leachates. On the other hand, PP and PLA generally displayed minimal to no toxicity in the studied species. Estimated species sensitivity distribution curves (SSD) show that PHBv leachates can be 10 times more hazardous to marine plankton than PP or PLA leachates, as demonstrated by the calculated Hazardous Concentration for 5 % of species (HC5). Qualitative chemical analysis supports the toxicological results, with 80 % of compounds being identified in PHBv leachates of which 2,4,6-trichlorophenol is worth mentioning due to the deleterious effects to aquatic biota described in literature. These findings underscore the fact that whereas environmental persistence can be targeted using PBP, the issue of chemical safety remains unsolved by some alternatives, such as PHBv. Gaining a comprehensive understanding of the toxicity profiles of PBP materials through a priori toxicological risk assessment is vital for their responsible application as alternatives to conventional plastics.

Polylactic acid nanoplastics (PLA-NPLs) induce adverse effects on an in vitro model of the human lung epithelium: The Calu-3 air-liquid interface (ALI) barrier.

The expected increments in the production/use of bioplastics, as an alternative to petroleum-based plastics, require a deep understanding of their potential environmental and health hazards, mainly as nanoplastics (NPLs). Since one important exposure route to NPLs is through inhalation, this study aims to determine the fate and effects of true-to-life polylactic acid nanoplastics (PLA-NPLs), using the in vitro Calu-3 model of bronchial epithelium, under air-liquid interphase exposure conditions. To determine the harmful effects of PLA-NPLs in a more realistic scenario, both acute (24 h) and long-term (1 and 2 weeks) exposures were used. Flow cytometry results indicated that PLA-NPLs internalized easily in the barrier (∼10 % at 24 h and ∼40 % after 2 weeks), which affected the expression of tight-junctions formation (∼50 % less vs control) and the mucus secretion (∼50 % more vs control), both measured by immunostaining. Interestingly, significant genotoxic effects (DNA breaks) were detected by using the comet assay, with long-term effects being more marked than acute ones (7.01 vs 4.54 % of DNA damage). When an array of cellular proteins including cytokines, chemokines, and growth factors were used, a significant over-expression was mainly found in long-term exposures (∼20 proteins vs 5 proteins after acute exposure). Overall, these results described the potential hazards posed by PLA-NPLs, under relevant long-term exposure scenarios, highlighting the advantages of the model used to study bronchial epithelium tissue damage, and signaling endpoints related to inflammation.

Characterization, hazard identification, and risk assessment of volatile organic compounds in Poly(butylene adipate-co-terephthalate)-based food contact articles.

The chemical safety of poly (butylene adipate-co-terephthalate) (PBAT) based food contact articles (FCAs) has aroused increasing toxicological concerns in recent years, but the chemical characterization and associated risk assessment still remain inadequate as it fails to elucidate the distribution pattern and discern the potential genotoxic and carcinogenic hazards of the identified substances. Herein, the volatile organic compounds (VOCs) in 50 batches of PBAT-based FCAs of representative categories and 10 batches of PLA and PBAT pellets were characterized, by which 237 VOCs of 10 chemical categories were identified and exhibited characteristic distribution patterns in the chemical spaces derived from their molecular descriptors. Chemical hazards associated with the identified VOCs were discerned by a hazard-driven classification scheme integrating hazard-related knowledge from multiple publicly available sources, and 34 VOCs were found to bear genotoxic or carcinogenic hazards and to feature higher average molecular weight than the other VOCs. Finally, the Risk and hazard quotient (HQ) calculated as the metrics of risk suggested that all identified VOCs posed acceptable risks (Risk<10-4 or HQ < 1), whereas oxolane, butyrolactone, N,N-dimethylacetamide, 2-butoxyethanol, benzyl alcohol, and 1,2,3-trichloropropane posed non-negligible (Risk>10-6) genotoxic or carcinogenic risk and thus should be of prioritized concern to promote the chemical safety of PBAT-based FCAs.

Multi-level toxicity assessment of polylactic acid (PLA) microplastics on the cladoceran Daphnia magna.

The accumulation of plastics waste in the environment has raised a worrisome concern, moving the society to seek out for sustainable solutions, such as the transition from the use of fossil-based, conventional plastics to bioplastics (BPs). However, once in the environment bioplastics have the same probability to accumulate and experience weathering processes than conventional plastics, leading to the formation of microplastics (MPs). However, to date the information on the potential toxicity of MPs originated from the weathering of bioplastics is limited. Thus, this study aimed at investigating the adverse effects induced by the exposure to MPs made of a bioplastic polymer, the polylactic acid (PLA), towards the freshwater cladoceran Daphnia magna. Organisms were exposed for 21 days to three concentrations (0.125 µg/mL, 1.25 µg/mL and 12.5 µg/mL) of PLA microplastics (hereafter PLA-MPs). A multi-level approach was performed to investigate the potential effects through the biological hierarchy, starting from the sub-individual up to the individual level. At the sub-individual level, changes in the oxidative status (i.e., the amount of reactive oxygen species and the activity of antioxidant and detoxifying enzymes) and oxidative damage (i.e., lipid peroxidation) were explored. Moreover, the total caloric content as well as the content of protein, carbohydrate and lipid content assess were used to investigate the effects on energy reserves. At individual level the changes in swimming activity (i.e., distance moved and swimming speed) were assessed. Our results showed that the exposure to PLA-MPs induced a slight modulation in the oxidative status and energy reserves, leading to an increase in swimming behavior of treated individuals compared to control conspecifics. These results suggest that the exposure to MPs made of a bioplastic polymer can induce adverse effects similar to those caused by conventional polymers.

Biodegradable microplastics: Uptake by and effects on the rockpool shrimp Palaemon elegans (Crustacea: Decapoda).

Ingestion of microplastics can lead to deleterious consequences for organisms, as documented by numerous laboratory studies. The current knowledge is based on a multitude of effect studies, conducted with conventional fossil-based and non-degradable plastics. However, there is a lack of information about the acceptance and the effects of novel bio-based and biodegradable plastics. Biodegradable plastics are considered an alternative to conventional plastics and are showing rapidly growing production rates. Biodegradable plastics can disperse into the environment in the same way as conventional plastics do, becoming available to marine organisms. This study aims to provide new insights into the uptake and effects of biodegradable microplastics on marine invertebrates. Rockpool shrimp, Palaemon elegans, were fed with algal flakes coated with polylactic acid (PLA), polyhydroxybutyrate-co-valerate (PHBV) and conventional low-density polyethylene (LDPE) microparticles. Live observations showed that all of the different types of microplastics were ingested. After dissection of the shrimp, less LDPE particles were found in the stomachs than PLA and PHBV particles. This indicates a longer retention time of biodegradable microplastics compared to conventional microplastics. Presumably, less LDPE particles were ingested or evacuated from the stomach, probably by regurgitation. The ingestion of microparticles of all types of plastics induced enzymatic activity of short-chain carboxylesterases in the midgut glands of the shrimp. However, only PLA induced enzymatic activity of medium-chain carboxylesterases. Palaemon elegans showed no oxidative stress response after ingestion of microparticles, irrespective of polymer type. From our results we conclude that biodegradable plastics might have different effects than conventional plastics. The longer retention times of biodegradable plastics might enhance exposure to leaching additives and other harmful substances. Our study provides new insights into how biodegradable plastics might affect aquatic fauna and indicate that the use of biodegradable plastics needs to be reconsidered to some extent.

Photoaging of biodegradable nanoplastics regulates their toxicity to aquatic insects (Chironomus kiinensis) by impairing gut and disrupting intestinal microbiota.

Biodegradable plastic, a widely used ecofriendly alternative to conventional plastic, easily form nanoplastics (NPs) upon environmental weathering. However, the effects and underlying mechanisms governing the toxicity of photoaged biodegradable NPs to aquatic insects are not understood. In this study, we investigated the photoaging of polylactic acid nanoplastics (PLA-NPs, a typical biodegradable plastic) that were placed under xenon arc lamp for 50 days and 100 days and compared the toxicity of virgin and photoaged PLA-NPs to Chironomus kiinensis (a dominant aquatic insect). The results showed that photoaged PLA-NPs significantly decreased the body weight, body length and emergence rate of C. kiinensis. Additionally, photoaged PLA-NPs induced more severe gut oxidative stress, histological damage, and inflammatory responses than virgin PLA-NPs. Furthermore, the alpha diversity of gut microbiota was lower in photoaged PLA-NPs group than virgin PLA-NPs. The relative abundance of key gut bacteria related to intestinal barrier defense, immunity, and nutrient absorption was reduced more significantly in photoaged PLA-NPs group than virgin PLA, indirectly leading to stronger gut damage and growth reduction. A stronger impact of photoaged PLA-NPs on the gut and its microbiota occurred because photoaging reduced the size of NPs from 255.5 nm (virgin PLA) to 217.1 nm (PLA-50) and 182.5 nm (PLA-100), induced surface oxidation and enhancement of oxidative potential, and improved the stability of NPs, thereby exacerbating toxicity on the gut and its microbiota. This study provides insights into the effects of biodegradable NPs on aquatic insects and highlights the importance of considering biodegradable nanoplastic aging in risk assessments.

Are bioplastics safe? Hazardous effects of polylactic acid (PLA) nanoplastics in Drosophila.

The expanded uses of bioplastics require understanding the potential health risks associated with their exposure. To address this issue, Drosophila melanogaster as a versatile terrestrial in vivo model was employed, and polylactic acid nanoplastics (PLA-NPLs), as a proxy for bioplastics, were tested as a material model. Effects were determined in larvae exposed for 4 days to different concentrations (25, 100, and 400 µg/mL) of 463.9 ± 129.4 nm PLA-NPLs. Transmission electron microscopy (TEM) and scanning electron microscope (SEM) approaches permitted the detection of PLA-NPLs in the midgut lumen of Drosophila larvae, interacting with symbiotic bacteria. Enzymatic vacuoles were observed as carriers, collecting PLA-NPLs and enabling the crossing of the peritrophic membrane, finally internalizing into enterocytes. Although no toxic effects were observed in egg-to-adult survival, cell uptake of PLA-NPLs causes cytological disturbances and the formation of large vacuoles. The translocation across the intestinal barrier was demonstrated by their presence in the hemolymph. PLA-NPL exposure triggered intestinal damage, oxidative stress, DNA damage, and inflammation responses, as evaluated via a wide set of marker genes. Collectively, these structural and molecular interferences caused by PLA-NPLs generated high levels of oxidative stress and DNA damage in the hemocytes of Drosophila larvae. The observed effects point out the need for further studies aiming to deepen the health risks of bioplastics before adopting their uses as a safe plastic alternative.

Identification of non-volatile non-intentionally added substances from polyester food contact coatings and genotoxicity assessment of polyester coating's migrates.

Can's polyester coatings are intended to replace epoxy-phenolic ones due to rising safety concern regarding the potential release of bisphenol A under increased regulations and consumer pressure. In this study, hazard linked to the migration of non-intentionally added substances from a single polyester-coated tin plate (5 batches) to canned food has been studied. Migration tests were performed using acetonitrile (ACN) and ethanol (EtOH) 95 %. Non-targeted analyses by liquid chromatography-high-resolution mass spectrometry revealed the presence of four cyclic oligoesters classified as Cramer class III substances with an estimated exposure (calculated for French population only) below the threshold of toxicological concern value of 1.5 µg/kg b.w./day, suggesting a no safety concern. Moreover, migrates were tested using in vitro genotoxicity DNA damage response (DDR) test and mini mutagenicity test (MMT) with different strains of S. Typhimurium using direct incorporation (TA100, TA98, TA102, TA1537) and pre-incubation (TA100, TA98) methods. Samples were negative in both bioassays suggesting the absence of genotoxicity/mutagenicity of the mixtures. To verify any false negative response due to matrix effect, migrates were spiked with corresponding positive controls in parallel with the MMT and the DDR test. No matrix effect was observed in these experimental conditions.

Plastic food? Energy compensation of zebrafish (Danio rerio) after long-term exposure to polylactic acid biomicroplastics.

The extensive use of bio-based plastics has led to their widespread distribution in the environment. However, their long-term ecological impact on aquatic animals is not well understood. In this study, adult zebrafish (Danio rerio) were exposed to 1000 items·L-1 of either polylactic acid (PLA) or polyethylene terephthalate (PET) microplastics (MPs), for 90 days. PLA is a typical bio-based plastic, while PET is a typical petroleum-derived plastic. The abundances of PLA and PET MPs in fish intestines were 981 ± 66 and 671 ± 151 items per fish, respectively, indicating a greater amount of PLA MP residues than PET MPs. However, the inhibitory effect of PET on fish weight was 1.8 times higher than that of PLA, suggesting energy compensation in PLA-treated zebrafish. Proliferation of Lactobacillus was observed in the fish intestines of the PLA group, indicating increased utilization capacity of intestinal flora for lactic acid production during PLA degradation. Metabolomics showed that the tricarboxylic acid pathway was up-regulated in the PLA group compared with that in the PET group, providing evidence of energy compensation. However, more ingested PLA MPs caused more significant histological damage to fish intestines than PET MPs. Therefore, the ecological risks of bio-based plastics still require attention.

Revealing the environmental hazard posed by biodegradable microplastics in aquatic ecosystems: An investigation of polylactic acid's effects on Microcystis aeruginosa.

The influence of petroleum-based microplastics (MPs) on phytoplankton has been extensively studied, while research on the impact of biodegradable MPs, derived from alternative plastics to contest the environmental crisis, remains limited. This study performed a 63 days co-incubation experiment to assess the effect of polylactic acid MPs (PLA-MPs) on the growth, physiology, and carbon utilization of M. aeruginosa and the change in PLA-MPs surface properties. The results showed that despite PLA-MPs induced oxidative stress and caused membrane damage in M. aeruginosa, the presence of PLA-MPs (10, 50, and 200 mg/L) triggered significant increases (p < 0.05) in the density of M. aeruginosa after 63 days. Specifically, the algal densities upon 50 and 200 mg/L PLA-MPs exposure were increased by 20.91% and 36.31% relative to the control, respectively. Meanhwhile, the reduced C/O ratio on PLA-MPs surface and change in PLA-MPs morphological characterization, which is responsible for substantially increase in the aquatic dissolved inorganic carbon concentration during the co-incubation, implying the degradation of PLA-MPs; thus, provided sufficient carbon resources that M. aeruginosa could assimilate. This was in line with the declined intracellular carbonic anhydrase content in M. aeruginosa. This study is the first attempt to uncover the interaction between PLA-MPs and M. aeruginosa, and the finding that their interaction promotes the degrading of PLA-MPs meanwhile favoring M. aeruginosa growth will help elucidate the potential risk of biodegradable MPs in aquatic environment.

Polylactic Acid Microplastics Do Not Exhibit Lower Biological Toxicity in Growing Mice Compared to Polyvinyl Chloride Microplastics.

Biomicroplastics (BMPs) will be produced during bioplastic degradation (i.e., polylactic acid), although bioplastics have been widely used for food packaging. Like conventional microplastics (MPs), BMPs would be mistakenly ingested into the body through diet or drinking water, but their health risks in vivo are poorly understood. Here, we deeply compared the toxicity difference between irregularly shaped polylactic acid (PLA-MPs, 16-350 µm) and polyvinyl chloride (PVC-MPs, 40-300 µm) MPs in growing mice. After six weeks of exposure, PLA-MP exposure resulted in more severe inhibition of the mice's weight gain than PVC-MPs did. Both PLA- and PVC-MPs significantly elevated the levels of oxidative stress. Moreover, significant changes including altered transcriptional profiles and significantly differentially expressed genes in liver and colon transcription levels were observed in the PVC- and PLA-MP groups. Compared with PVC-MPs, PLA-MPs have a stronger effect on lipid metabolism and digestive systems. PLA-MPs also caused gut microbiota dysbiosis, significantly interfering with the relative abundance of microbiota and altering microbial diversity. These findings indicated the toxicities of PLA-MPs in growing mice were not significantly reduced compared to PVC-MPs, which would also provide new insights for re-examining bioplastic safety.

Comparative study on the toxic effects of secondary nanoplastics from biodegradable and conventional plastics on Streptomyces coelicolor M145.

Because of the excellent properties, plastics have been widely used in the past decades and caused serious environmental issues. As an excellent substitute for conventional plastics, the biodegradable plastics have attracted increasing attention. However, biodegradable plastics may produce more micro/nanoplastics in the short time compared with conventional plastics, and cause more serious ecological risks. In this study, the short-term toxicity of nanoplastics released from biodegradable and conventional plastics on Streptomyces coelicolor M145 was investigated. After 30 days of degradation, the biodegradable microplastics, polylactic acid (PLA) and polyhydroxyalkanoates (PHA) released more secondary nanoplastics than conventional microplastics, polystyrene (PS). After exposure, PLA and PHA nanoplastics showed significant toxicity to M145. The survival rate of M145 cells was 16.1% after treatment with PLA nanoplastics for 7 days (PLA-7). The toxicity of PHA was lower than that of PLA. This might have been due to the agglomeration of PHA nanoplastics in the solution. Compared with the controls, the PS secondary nanoplastics showed no significant toxicity to M145. After the treatment, the production of antibiotics, actinorhodin (ACT) and undecylprodigiosin (RED), significantly increased. The yields of ACT and RED reached their maximum values after treatment with PLA-7, which were 4.2-fold and 2.1-fold higher than those of the controls, respectively. The addition of biodegradable nanoplastics significantly increased the expression of these key pathway-specific regulatory genes, leading to increased antibiotic production. This study provides toxicological insights into the impacts of conventional and biodegradable microplastics on S. coelicolor.

The effects of polylactic acid bioplastic exposure on midgut microbiota and metabolite profiles in silkworm (Bombyx mori): An integrated multi-omics analysis.

Polylactic acid (PLA) is a highly common biodegradable plastic and a potential threat to health and the environment. However, limited data are available on the effects of PLA exposure in the silkworm (Bombyx mori), a model organism used in toxicity studies. In this study, silkworms with or without PLA exposure (P1: 1 mg/L, P5: 5 mg/L, P25: 25 mg/L, and P0: 0 mg/L) for the entire 5th instar period were used to investigate the impact of PLA exposure on midgut morphology, larvae growth, and survival. Mitochondrial damage was observed in the P5 and P25 groups. The weights of the P25 posterior silk gland (5th day in the 5th instar), mature larvae and pupae were all significantly lower than those of the controls (P < 0.05). Dead worm cocoon rates and larva-pupa to 5th instar larvae ratios showed a positive and negative dose-dependent manner with respect to PLA concentrations, respectively. Additionally, reactive oxygen species levels and superoxide dismutase activity of the P25 midgut were significantly higher and lower when compared with controls, respectively (P < 0.05). The molecular mechanisms underlying the effects of PLA and associated physiological responses were also investigated. In the midgut metabolome, 127 significantly different metabolites (variable importance projection >1 and P < 0.05) were identified between the P0 and P25 groups and were mainly enriched for amino acid metabolism and energy supply pathways. The 16 S rDNA data showed that PLA altered microbial richness and structural composition. Microbiota, classified into 34 genera and 63 species, were significantly altered after 25 mg/L PLA exposure (P < 0.05). Spearman's correlation results showed that Bifidobacterium catenulatum and Schaalia odontolytica played potentially vital roles during exposure, as they demonstrated stronger correlations with the significantly different metabolites than other bacterial species. In sum, PLA induced toxic effects on silkworms, especially on energy- and protein-relevant metabolism, but at high concentrations (25 mg/L). This prospective mechanistic investigation on the effects of PLA on larval toxicity provides novel insight regarding the ecological risks of biodegradable plastics in the environment.

Effects of polycaprolactone degradation products on the water flea, Daphnia magna: Carbodiimide additives have acute and chronic toxicity.

Plastics have benefited our lives in many ways, but their long persistence in the environment causes serious problems. Rapid decomposition and detoxification of plastics after use are significant challenges. As a possible solution, biodegradable plastics have attracted attention, and for environmental risk assessment research on polymer toxicity, use of indicator organisms, like water fleas and fish, has increased globally. However, such research often focuses on standardized substances without considering changes in toxicity due to plastic degradation products. Additionally, tests generally focus on acute toxicity, while long-term effects on organismal reproduction and lifespan are largely unknown. Understanding the impact of degraded polymers on biological activities is crucial for accurate risk assessment. In this study, we investigated the biological toxicity of substances generated during degradation of polycaprolactone (PCL), a common biodegradable plastic, using the indicator organism, Daphnia magna. We examined PCL, oligocaprolactones (OCLs), and monomers resulting from polymer cleavage, as well as carbodiimides, added during polyester synthesis. As a result, PCL, which is insoluble in water, reduced individual survival and total number of offspring at an exposure concentration of 100 mg/L, while no toxicity was observed for water-soluble degradation products, OCLs, and monomers. Furthermore, carbodiimides, which are expected to be released during PCL degradation, showed strong toxicity, significantly reducing individual survival and total number of offspring at 0.1-10 mg/L. These findings suggest that changes in physical properties due to polymer degradation and release of additives can significantly alter their toxicity.

Effect of bio-based microplastics on earthworms Eisenia andrei.

The contribution of bio-based plastics in the global market is gradually growing and diversifying. Therefore, it is necessary to assess their environmental impact including the biotic parts of ecosystems. Earthworms are regarded as functionally essential and useful bioindicators of ecological disturbances in the terrestrial ecosystems. The purpose of this study was to evaluate the impact of three innovative bio-based plastics on earthworms Eisenia andrei in the long-term experiments. It comprised the mortality, body mass and reproduction ability of earthworms as well as the oxidative stress response. Regarding the latter the activities of catalase (CAT) and superoxide dismutase (SOD) involved in the antioxidant system of earthworms were determined. Two out of three bio-based materials tested were polylactic acid-based (PLA-based) plastics, while one was poly(hydroxybutyrate-co-valerate)-based (PHBV-based) plastic. Neither mortality nor weight of adult earthworms was affected even at high concentration of the bio-based plastics up to 12.5 % w/w in the soil. Reproduction ability occurred to be more sensitive endpoint than mortality or body mass. At the concentration of 12.5 % w/w each of the studied bio-based plastics contributed to the decrease of the earthworm reproduction at statistically significant level. PLA-based plastics exerted stronger effect on earthworm reproduction ability than PHBV-based plastic did. CAT activity turned out to be a good indicator of the cellular response against oxidative stress induced by bio-based plastics in earthworms. The activity of this enzyme increased in the response to the exposure to the bio-based plastics compared to the level achieved in the control tests. It was from 16 % to about 84 % dependent on the material tested and its concentration in the soil. Finally, the reproduction ability and catalase activity are recommended to be used in the evaluation of the potential impacts of bio-based plastics on earthworms.

Sublethal effects of bio-plastic microparticles and their components on the behaviour of Daphnia magna.

Bioplastics arise as an alternative to plastic production delinked from fossil resources. However, as their demand is increasing, there is a need to investigate their environmental fingerprint. Here we study the toxicity of microplastics (MPLs) of two widely used materials, the polylactic acid (PLA) and the polyhydroxybutyrate (PHB) on the environmental aquatic model species Daphnia magna. The study was focused on sublethal behavioural and feeding endpoints linked to antipredator scape responses and food intake. The study aimed to test that MPLs from single-use household comercial items and among them bioplastics should be more toxic than those obtained from standard plastic polymers and fossil plastic materials due to the greater amount of plastic additives, and that MPLs should be more toxic than plastic extracts due to the contribution of both particle and plastic additive toxicity. MPLs were obtained by cryogenic grinding and sea-sand erosion to obtain irregular particles. MPL included standard polymers and nine comercial items of PLA and PHB and one fossil-based material of high-density polyethylene (HDPE). The additive content in commercial items was characterised by liquid chromatography coupled with high-resolution mass spectrometry. D. magna juveniles were exposed for 24 h to particles and their plastic extracts. Results indicated that the toxicity of bioplastic particles was five times higher than the effects produced by exposure to the content of the additives alone, that bioplastic particles were more toxic than fossil ones and that particles obtained from commercial items were more toxic than those obtained from PLA, PHB or HDPE polymer standards. Predicted toxicity from the measured plastic additives in the studied commercially available household items, however, was poorly related with the observed behavioural and feeding effects. Further research on unknown chemical components together with physical factors is need it to fully understand the mechanisms of toxicity of bioplastic materials.

Effect of polylactic acid microplastics and lead on the growth and physiological characteristics of buckwheat.

Microplastics (MPs) and heavy metals are common, often co-existing pollutants, that threaten crop growth and productivity worldwide. We analysed the adsorption of lead ions (Pb2+) to polylactic acid MPs (PLA-MPs) and their single factor and combined effects on tartary buckwheat (Fagopyrum tataricum L. Gaertn.) in hydroponics by measuring changes in the growth characteristics, antioxidant enzyme activities and Pb2+ uptake of buckwheat in response to PLA-MPs and Pb2+. PLA-MPs adsorbed Pb2+, and the better fitting second-order adsorption model implied that Pb2+ was adsorbed by chemisorption. However, the similar Pb2+ contents in the plants treated with Pb2+ only and those treated with the combined PLA-MPs-Pb2+ suggested that the adsorption played no role in the uptake of Pb2+. Low concentrations of PLA-MPs promoted shoot length. At high concentrations of both PLA-MPs and Pb2+, buckwheat growth was inhibited, and leaf peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) contents were higher than in the control. No significant differences were observed in seedling growth between exposure to Pb2+ only and combined exposure to PLA-MPs with Pb2+, implying that PLA-MPs did not increase the toxicity of Pb2+ at macroscopic level. POD activity was higher and chlorophyll content was lower with PLA-MPs in the low Pb2+ dose treatments, suggesting that PLA-MPs may increase the toxicity of naturally occurring Pb2+. However, the conclusions must be verified in controlled experiments in natural soil conditions over the whole cultivation period of buckwheat.

Polylactic acid microparticles in the range of µg/L reduce reproductive capacity by affecting the gonad development and the germline apoptosis in Caenorhabditis elegans.

Polylactic acid (PLA) accounts for approximately 45% of the global market of biodegradable plastics. Using Caenorhabditis elegans as an animal model, we examined the effect of long-term exposure to PLA microplastic (MP) on reproductive capacity and the underlying mechanism. Brood size, number of fertilized eggs in uterus, and number of hatched eggs were significantly reduced by exposure to 10 and 100 µg/L PLA MP. Number of mitotic cells per gonad, area of gonad arm, and length of gonad arm were further significantly decreased by exposure to 10 and 100 µg/L PLA MP. In addition, exposure to 10 and 100 µg/L PLA MP enhanced germline apoptosis in the gonad. Accompanied with the enhancement in germline apoptosis, exposure to 10 and 100 µg/L PLA MP decreased expression of ced-9 and increased expressions of ced-3, ced-4, and egl-1. Moreover, the induction of germline apoptosis in PLA MP exposed nematodes was suppressed by RNAi of ced-3, ced-4, and egl-1, and strengthened by RNAi of ced-9. Meanwhile, we did not detect the obvious effect of leachate of 10 and 100 µg/L PLA MPs on reproductive capacity, gonad development, germline apoptosis, and expression of apoptosis related genes. Therefore, exposure to 10 and 100 µg/L PLA MPs potentially reduces the reproductive capacity by influencing the gonad development and enhancing the germline apoptosis in nematodes.