Invertebrates, including crustaceans, rely on cellular and humoral immune responses to protect against extrinsic and intrinsic factors that threaten their integrity. Recently, different immune ...parameters have been increasingly used as biomarkers of effects of pollutants and environmental change. Here, we describe the dynamics of the innate immune response of the terrestrial crustacean
Porcellio scaber
to injection of a single dose of lipopolysaccharide (LPS), an important molecular surface component of the outer membrane of Gram-negative bacteria. The aim was to provide a basis for interpretation of change in immune parameters as a result of different challenges, including microplastics and nanoplastics exposure. Changes in total and differential numbers of hemocytes, hemocyte viability, and humoral immune parameters (i.e., phenoloxidase-like activity, nitric oxide levels) were assessed at different times (3, 6, 12, 24, 48 h). An injection of 0.5 μg/μL LPS into the body of
P. scaber
resulted in a rapid decrease (3 h after LPS injection) in the total number of hemocytes and reduced viability of the hemocytes. This was accompanied by changed proportions of the different hemocyte types, as a decrease in the numbers of semigranulocytes and granulocytes, and a marked increase in the numbers of hyalinocytes. In addition, phenoloxidase-like activity and nitric oxide levels in the hemolymph were increased at 3 h and 6 h, respectively, after the LPS challenge. Forty-eight hours after LPS injection, the immune parameters in the hemolymph of
P. scaber
had returned to those before the LPS challenge. This suggests that the innate immune system successfully protected
P. scaber
from the deleterious effects of the LPS challenge. These data indicate the need to consider the dynamics of innate immune responses of
P. scaber
when effects of infections, pollutants, or environmental changes are studied. We also propose an approach to test the immunocompetence of organisms after different challenges in ecotoxicity studies, based on the dynamics of their immune responses.
The presence of microplastics (MPs) in the environment has raised many concerns, and therefore approaches and technologies to remove them in situ are of high interest. In this context, we ...investigated the interactions between polyethylene MPs (fragments with a mean size of 149 ± 75 μm) and an aquatic floating macrophyte Lemna minor in order to assess its potential use for in situ phytoremediation. We first investigated the long-term effects of a high (100 mg/L = 9600 MPs/L), but still environmentally relevant concentration of MPs on L. minor. Subsequently bioadhesion of MPs was studied and the number and strength of MPs adhering to plant biomass were assessed. MPs did not adversely affect various parameters of plants (e.g., specific growth rate, chlorophyll contents, total antioxidant capacity, electron transport system activity, and contents of energy-rich molecules) throughout the duration of the experiment (12 weeks), except for the first week of the experiment, when protein content and total antioxidant capacity were affected. On the other hand, MPs affected the root length of L. minor during the first eight weeks of the experiment, while further exposure resulted in a decrease in the effects, indicating the ability of L. minor to tolerate the presence of MPs for a long period of time. MPs adhered rapidly to the plant biomass and the average percentages of strongly and weakly adhered particles were 6.5% and 20.0%, respectively, of the total MPs applied. In summary, results of this study suggest that L. minor can tolerate hotspot concentrations of MPs and can collect MPs from the water surface. Therefore, phytoremediation using floating plants could be considered as a potential method for in situ removal of MPs from the aquatic environment.
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•Microplastics (MPs) adhere to the biomass of Lemna minor.•MPs have no significant effect on Lemna minor growth over a 12-week period.•6.5% and 20.0% of all MPs are strongly and weekly adhered, respectively.•Phytoremediation could be used to remove MPs from the aquatic environment.
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•The effects of polyester fibers on soil animals were studied for the first time.•Enchytraeid reproduction decreased up to 30% but only by long fibers in soil.•Isopod energy reserves ...and feeding activity were affected by fibers in soil.•Polyester fibers were not very harmful to soil invertebrates in 21–28-days exposure.•Polyester fibers can enter terrestrial food webs by ingestion by soil invertebrates.
Polyester fiber is one of the most abundant types of microplastics in the environment. A major proportion of the fibers entering wastewater treatment plants end up in sewage sludge, which is used as a soil fertilizer in many countries. As their impacts in the terrestrial environment are still poorly understood, we studied the effects of polyester fibers on enchytraeids (Enchytraeus crypticus), springtails (Folsomia candida), isopods (Porcellio scaber) and oribatid mites (Oppia nitens), all playing an important role in soil decomposer food webs. We exposed these invertebrates in the laboratory to short (12 µm–2.87 mm) and long (4–24 mm) polyester fibers, spiked in soil or in food at five concentrations ranging from 0.02% to 1.5% (w/w) and using five replicates. Overall the effects of polyester fibers on the soil invertebrates were slight. Energy reserves of the isopods were slightly affected by both fiber types, and enchytraeid reproduction decreased up to 30% with increasing fiber concentration, but only for long fibers in soil. The low ingestion of long fibers by the enchytraeids suggests that this negative impact arose from a physical harm outside the organism, or from indirect effects resulting from changes in environmental conditions. The short fibers were clearly ingested by enchytraeids and isopods, with the rate of ingestion positively related to fiber concentration in the soil. This study shows that polyester fibers are not very harmful to soil invertebrates upon short-term exposure. However, longer lasting, multigeneration studies with functional endpoints are needed to reveal the possible long-term effects on soil invertebrates and their role in the decomposition process. This study also shows that polyester fibers can enter terrestrial food web via ingestion of fibers by soil invertebrates.
Mulching films, widely used in agriculture, are a large source of microplastics (MPs) to soil. However, there is little knowledge on the long-term effects of agricultural MPs on soil invertebrates. ...We investigated the effects of MPs from conventional non-biodegradable, fossil-based, low-density polyethylene (PE) and biodegradable fossil-based poly(butylene adipate-coterephthalate) (starch-PBAT blend) mulching films on two generations of the mealworm Tenebrio molitor. No effects of MPs (0.005 %–5 %, w/w dry food) on mealworm development and survival were observed until the end of the experiments (12 weeks for the first generation, nine weeks for the second generation), but effects on their moulting and growth were observed. These were most evident for PE MPs (5 %, w/w), where a decrease in larval growth and moulting was noted in the first generation. On the contrary, PBAT MPs (5 %, w/w) significantly induced the growth of mealworms in the second generation. In addition, there was a non-significant trend towards increased growth at all other PBAT MP exposure concentrations. Increased growth is most likely due to the biodegradation of starch PBAT MPs by mealworms. Overall, these data suggest that PE and PBAT MPs do not induce significant effects on mealworms at environmentally relevant concentrations, but rather only at very high exposure concentrations (5 %).
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•PBAT and PE MPs up to 5 % (w/w) do not affect the survival and development of mealworms.•Larval moult and growth are decreased in PE MP exposure (5 %) in the 1st generation.•Larval growth is significantly increased in PBAT MP exposure (5 %) in the 2nd generation.•PE and PBAT MPs do not induce significant effects on mealworms at environmental levels.
The COVID-19 pandemic has increased the use of disposable plastics, including medical masks, which have become a necessity in our daily lives. As these are often improperly disposed of, they ...represent an important potential source of microplastics in the environment. We prepared microplastics from polypropylene medical masks and characterised their size, shape, organic chemical leaching, and acute toxicity to the planktonic crustacean
Daphnia magna
. The three layers of the masks were separately milled and characterised. Each of the inner frontal, middle filtering, and outer layers yielded different types of microplastics: fibres were obtained from the inner and outer layer, but irregular fragments from the middle layer. The shape of the obtained microplastics differed from the initial fibrous structure of the intact medical mask layers, which indicates that the material is deformed during cryo-milling. The chemical compositions of plastics-associated chemicals also varied between the different layers. Typically, the inner layer contained more chemicals related to antimicrobial function and flavouring. The other two layers also contained antioxidants and their degradation products, plasticisers, cross-linking agents, antistatic agents, lubricants, and non-ionic surfactants. An acute study with
D. magna
showed that these microplastics do not cause immobility but do physically interact with the daphnids. Further long-term studies with these microplastics are needed using a suite of test organisms. Indeed, studies with other polypropylene microplastics have shown numerous adverse effects on other organisms at concentrations that have already been reported in the environment. Further efforts should be made to investigate the environmental hazards of polypropylene microplastics from medical masks and how to handle this new source of environmental burden.
Tire particles pose a potential threat to terrestrial organisms because they are deposited in large quantities in the soil by tire wear abrasion, and moreover their chemical complexity poses an ...additional risk. Microplastics can affect several physiological processes in organisms, including those related to immunity. Therefore, we investigated the expression profile of selected immune-related genes (MnSod, Manganese Superoxide dismutase; Cat, Catalase; CypG, Cyclophilin G; Nos, Nitric oxide synthase; Ppae2a, Prophenoloxidase-activating enzyme 2a; Dscam, Down syndrome cell adhesion molecule; Myd88, Myeloid-differentiation factor 88; Toll4, Toll-like receptor 4; Mas-like, Masquerade-like protein) in haemocytes and the digestive gland hepatopancreas of terrestrial crustacean Porcellio scaber after two different time exposures (4 and 14 days) to tire particles in soil. Our results reveal for the first time the response of P. scaber after microplastic exposure at the transcriptome level. We observed time- and tissue-dependent changes in the expression of the analysed genes, with more pronounced alterations in haemocytes after 14 days of exposure. Some minor changes were also observed in hepatopancreas after 4 days. Changes in the expression profile of the analysed genes are a direct indication of a modulated immune status of the test organism, which, however, does not represent an adverse effect on the test organism under the given conditions. Nevertheless, the question remains whether the observed change in immune status affects the immunocompetence of the test organism.
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•Exposure to tire particles in soil impacts the expression of tested genes in woodlice.•Modulations in immune-related gene expression were time- and tissue-dependent.•The most striking changes were observed in haemocytes after 14 days.•Some genes were also changed in hepatopancreas but only after 4 days.
Microplastics are very common contaminants in the environment. Despite increasing efforts to assess the effects of microplastics on soil organisms, there remains a lack of knowledge on how organisms ...respond to diverse types of microplastics after different exposure durations. In the present study, we investigated the immune response of the terrestrial crustacean Porcellio scaber exposed to the two most common microplastic particles in the environment: polyester fibres and tyre particles. We also tested two natural particles: wood dust and silica powder, with all treatments performed at 1.5% w/w. The response of P. scaber was evaluated at the level of the immune system, and also the biochemical, organism and population level, after different exposure durations (1, 2, 4, 7, 14, 21 days). These data reveal dynamic changes in the levels of some immune parameters shortly after exposure, with a gradual return to control values. The total number of haemocytes was significantly decreased after 4 days of exposure to tyre particles, while the proportion of different haemocyte types in the haemolymph was altered shortly after exposure to both polyester fibres and tyre particles. Moreover, 7 days of exposure to tyre particles resulted in increased superoxide dismutase activity in the haemolymph, while metabolic activity in whole woodlice (measured as electron transport system activity) was increased after exposure for 7, 14 and 21 days. In contrast, the natural particles did not elicit any significant changes in the measured parameters. Survival and feeding of P. scaber were not altered by exposure to the microplastics and natural particles in soil. Overall, this study defines a time-dependent transient immune response of P. scaber, which indicates that immune parameters represent sensitive biomarkers of exposure to microplastics. We discuss the importance of using natural particles in studies of microplastics exposure and their effects.
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•We studied natural particles and microplastics for immune effects in Porcellio scaber•Haemocyte proportions in haemolymph rapidly and transiently changed for microplastics•Exposure to tyre particles significantly increased whole woodlouse metabolic activity•There were no changes in measured parameters for exposure to natural particles
Agricultural mulching films are potential sources of microplastics (MPs) in soil. As an alternative to conventional non-degradable mulching films, a variety of different biodegradable mulching films ...are used. However, it is not yet known whether MPs from biodegradable mulching films pose a lower risk to terrestrial invertebrates compared to MPs from conventional mulching films. In this study, the effects of MPs produced from two conventional polyethylene (PE-1 and PE-2) and two biodegradable (starch-based poly(butylene adipate co-terephthalate); PBAT-BD-1, and PBAT-BD-2) fossil-based mulching films on terrestrial crustacean woodlice Porcellio scaber and mealworm Tenebrio molitor were compared. A key finding was that no clear differences in induced responses between biodegradable and conventional MPs were detected. No adverse effects on P. scaber after two weeks and on T. molitor after four weeks of exposure were observed up to 5 % (w/w dry soil) of either MP type. However, some sublethal physiological changes in metabolic rate and immune parameters were found in P. scaber after two weeks of exposure indicating a response of organisms to the presence of MP exposure in soil. In addition, it was demonstrated that both types of MPs might affect the soil water holding capacity and pH. In conclusion, we confirmed that biodegradable MPs can induce responses in organisms hence further studies testing the environmental hazard of biodegradable MPs are justified.
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•PBAT and LDPE microplastics (MPs) induced various sublethal responses in woodlice.•Both types of MPs provoked an immune response in woodlice after 2 weeks-exposure.•Biodegradable starch-based and non-degradable conventional MPs induced similar responses.•MPs did not affect mealworm growth and survival after 4 weeks of exposure up to 5 %.
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