Crustacean Daphnia Magna Research Articles

Evaluation of Ecotoxicity of Wastewater from the Full-Scale Treatment Plants

In this work, the influence of wastewater from full-scale wastewater treatment plants (WWTPs) on aquatic and soil biota was reviewed and presented. Moreover, the methods and model organisms used in testing the ecotoxicity of wastewater were shown. It was found that wastewater usually affected the biochemical activity and growth of organisms such as bacteria, algae and protozoa. They contributed to the immobilization and death of inter alia crustaceans and fishes. The values of degree of inhibition or lethality widely varied dependent on the type of wastewater, the sampling point (influent or effluent) and the model organisms applied in the biotests. Thus, a battery of ecotoxicity tests using model organisms of different sensitivities should be employed. So far, bacteria (e.g., Vibrio fischeri), green microalgae (e.g., Raphidocelis subcapitata) and crustaceans (Daphnia magna) have been frequently used organisms in the biological assessment of wastewater. They were applied in almost half (bacteria) or more than half (microalgae, crustaceans) of papers analyzed in this study. In almost all studies, the reduction of wastewater toxicity after treatment processes was found. It was proven that the conventional activated sludge systems were efficient in the removal of wastewater toxicity from both municipal and industrial wastewater, while the tertiary stage of treatment, in particular chlorination or ozonation, contributed to the increase in wastewater toxicity.

CELF1 represses Doublesex1 expression via its 5' UTR in the crustacean Daphnia magna.

In sex determination of the crustacean Daphnia magna, male-specific expression of DM-domain transcription factor Doublesex1 (Dsx1) orchestrates the male developmental program triggered by environmental stimuli. We previously identified the CELF1 ortholog as a candidate of proteins associated with the 5’ UTR of the Dsx1α isoform. Here we report the CELF1-dependent suppression of Dsx1 expression in D. magna. During embryogenesis, CELF1 expression was not sexually dimorphic. Silencing of CELF1 led to the activation of Dsx1 expression both in female and male embryos. Overexpression of CELF1 in male embryos resulted in a reduction of Dsx1 expression. By these manipulations of CELF1 expression, the Dsx1 transcript level was not significantly changed. To investigate whether the CELF1 controls Dsx1 expression via its 5’ UTR, we injected the GFP reporter mRNA having intact Dsx1α 5’ UTR or mutated one lacking the GU-rich element (GRE) that is known as a binding site of the CELF1 ortholog. We found that deletion of the GRE significantly increased the reporter gene expression. These results indicate that CELF1 suppresses Dsx1 expression both in females and males, possibly at the post-transcriptional level. We speculate that CELF1 may avoid unintended Dsx1 expression and generation of sexual ambiguity by setting a threshold of Dsx1 expression.

Are natural deep eutectic solvents always a sustainable option? A bioassay-based study

The traditional use of organic solvents in various branches of industry is being rethought as these compounds very often display high volatility, toxicity and lipophilicity (related to the ability to interact with biological membranes). More recently, developments in the field of Green Chemistry are focusing on the design of more sustainable and cost-effective solvent alternatives like Ionic Liquids (ILs), bio-based solvents and natural deep eutectic solvents (NADESs). The present study aimed at performing an ecotoxicological screening of 15 NADESs using an extensive set of marine and freshwater bioassays, based on different endpoints as the following: immobilization of the crustacean Daphnia magna, growth inhibition of Raphidocelis subcapitata and of Phaeodactylum tricornutum, larval development alterations on the serpulid Ficopomatus enigmaticus and bioluminescence inhibition of Aliivibrio fischeri. What emerged was a general absence of toxicity of all samples. However, both algal assays showed a certain degree of biostimulation, up to over 100% growth increase in respect to controls with 8 out of 15 compounds tested with Raphidocelis subcapitata. Despite NADESs-induced negligible toxicity effects to invertebrates, encouraging their labelling as “sustainable” solvents, the liability of their intentional or accidental release into aquatic systems may represent a serious risk in terms of ecosystem functioning impairments.

Effects of Two Antiretroviral Drugs on the Crustacean Daphnia magna in River Water.

Antiretroviral (ARVs) drugs are used to manage the human immunodeficiency virus (HIV) disease and are increasingly being detected in the aquatic environment. However, little is known about their effects on non-target aquatic organisms. Here, Daphnia magna neonates were exposed to Efavirenz (EFV) and Tenofovir (TFV) ARVs at 62.5–1000 µg/L for 48 h in river water. The endpoints assessed were mortality, immobilization, and biochemical biomarkers (catalase (CAT), glutathione S-transferase (GST), and malondialdehyde (MDA)). No mortality was observed over 48 h. Concentration- and time-dependent immobilization was observed for both ARVs only at 250–1000 µg/L after 48 h, with significant immobilization observed for EFV compared to TFV. Results for biochemical responses demonstrated that both ARVs induced significant changes in CAT and GST activities, and MDA levels, with effects higher for EFV compared to TFV. Biochemical responses were indicative of oxidative stress alterations. Hence, both ARVs could potentially be toxic to D. magna.

The Daphnia carapace and other novel structures evolved via the cryptic persistence of serial homologs

Understanding how novel structures arise is a central question in evolution. Novel structures are often defined as structures that are not derived from (homologous to) any structure in the ancestor.1 The carapace of the crustacean Daphnia magna is a bivalved "cape" of exoskeleton. Shiga etal.2 proposed that the carapace of crustaceans like Daphnia and many other plate-like outgrowths in arthropods are novel structures that arose through the repeated co-option of genes like vestigial that also pattern insect wings.2-4 To determine whether the Daphnia carapace is a novel structure, we compare previous functional work2 with the expression of genes known to pattern the proximal leg region (pannier, araucan, and vestigial)5,6 between Daphnia, Parhyale, and Tribolium. Our results suggest that the Daphnia carapace did not arise by co-option but instead derived from an exite (lateral leg lobe) that emerges from an ancestral proximal leg segment that was incorporated into the Daphnia body wall. The Daphnia carapace, therefore, appears to be homologous to the Parhyale tergal plate and the insect wing.5 Remarkably, the vestigial-positive tissue that gives rise to the Daphnia carapace appears to be present in Parhyale7 and Tribolium as a small, inconspicuous protrusion. Thus, rather than a novel structure resulting from gene co-option, the Daphnia carapace appears to have arisen from a shared, ancestral tissue (morphogenetic field) that persists in a cryptic state in other arthropod lineages. Cryptic persistence of unrecognized serial homologs may thus be a general solution for the origin of novel structures.

Are "liquid plastics" a new environmental threat? The case of polyvinyl alcohol.

Despite the pollution induced by plastics become a well-known and documented problem, bringing many countries to adopt restrictions about their production, commercialization and use, the impact of another emerging category of synthetic polymers, represented by the Water-Soluble Polymers (WSPs), also known as "liquid plastics", is overlooked by scientific community. WSPs are produced in large quantities and used in a wide plethora of applications such as food packaging, pharmaceuticals and personal care products, cosmetics and detergents, with a consequent continuous release in the environment. The aim of this study was the investigation of the possible toxicity induced by polyvinyl alcohol (PVA), one of the main produced and used WSPs, on two freshwater model organisms, the crustacean Daphnia magna and the teleost Danio rerio (zebrafish). We evaluated the effects of solubilized standard PVA powder and PVA-based commercial bags for carp-fishing, at 3 different concentrations (1 µg/L, 0.5 mg/L and 1 mg/L), through the exposures for 14 days of D. magna (daphnids; age < 24 h) and for 5 days of zebrafish embryos (up to 120h post fertilization - hpf). As acute effects we evaluated the immobilization/mortality of specimens, while for chronic toxicity we selected several endpoints with a high ecological relevance, as the behavioural alteration on swimming performance, in real-time readout, and the activity of monoamine oxidase (MAO), a neuro-enzyme with a potential implication in the organism movement. The results showed the lack of significant effects induced by the selected substances, at all tested concentrations and in both model organisms. However, considering the wide plethora of available WSPs, other investigations are needed to provide the initial knowledge of risk assessment of these compounds contained in some consumer products.

Ecotoxicological Consequences of the Abatement of Contaminants of Emerging Concern by Ozonation—Does Mixture Complexity Matter?

Ozonation has been used to degrade persistent water contaminants, namely, pharmaceuticals and personal care products (PPCPs). However, ozonation can lead to by-products that can be more toxic than the parent compounds. This work aims to assess whether the ecotoxicological effects of ozonation are modified as the initial matrix being treated increases in complexity, considering mixtures of 2, 3, 4 and 5 PPCPs. The following PPCPs were used: two parabens (metylparaben (MP) and propylparaben (PP)), paracetamol (PCT), sulfamethoxazole (SMX) and carbamazepine (CBZ). The following model species were used to assess toxicity: the crustacean Daphnia magna, the microalgae Raphidocelis subcapitata, the macrophyte Lemna minor and the watercress Lepidium sativum. There was a trend of increased toxicity with increasing mixture complexity of the untreated samples, except for D. magna. The same was observed after ozonation with the exception of the mixture MP+PP, which showed high toxicity to all the tested species, namely 100% immobilization of D. magna. The toxicity of SMX to the primary producers decreased pronouncedly after ozonation, except for L. minor. This study highlights the importance of considering the complexity of the matrix being treated and of using an ecotoxicological test battery with a wide diversity of species for assessing ozonation efficiency.

The first plastic produced, but the latest studied in microplastics research: The assessment of leaching, ecotoxicity and bioadhesion of Bakelite microplastics

Bakelite, the first synthetic plastic, is a rather unexplored material in the field of ecotoxicology, despite its long production and use. The aim of this study was to investigate the ecotoxicity of Bakelite microplastics (before and after leaching) and its leachates on four aquatic organisms: the crustacean Daphnia magna, the plant Lemna minor, the bacterium Allivibrio fischeri and the alga Pseudokirchneriella subcapitata. Bakelite microplastics before and after leaching and leachates affected all organisms, but to varying degrees. Leachates showed increased ecotoxicity to Daphnia magna, while Pseudokirchneriella subcapitata was more affected by particles. For Lemna minor and Allivibrio fischeri, the effects of particles before leaching and leachate were comparable, while the negative effect of particles after leaching was minimal or not present. All leachates were analysed, and phenol and phenol-like compounds were the predominant organics found. In addition, bioadhesion of Bakelite microplastics to the surface of Daphnia magna and Lemna minor was confirmed, but the particles were mainly weakly adhered. Results of this study suggest that, in addition to the recently studied microplastics from consumer products (e.g. from polyethylene and polystyrene), microplastics from industrial plastics such as Bakelite may be of increasing concern, primarily due to leaching of toxic chemicals.

Protecting our environment, a motivating outdoor game for proteomics!

The global Earth ecosystem faces many intertwined threats, primarily anthropogenic pollution, drastic reduction of wild spaces, faster spread of pathogens, and global climate warming. Ecotoxicology, the integration of toxicology and ecology, aims to describe the effects of toxicants on organisms, whether at the level of the population, the community, the ecosystem, or the biosphere. Sentinel species are employed to assess threats to life, giving advance warning of danger. In this issue of Proteomics, Wilde and collaborators (Proteomics 2022, 22, https://doi.org/10.1002/pmic.202100289) present a comprehensive coverage of the proteome of the crustacean Daphnia magna, a species used to evaluate aquatic pollution. This study illustrates how current shotgun proteomics technology allows straightforward quantitation of any protein for whole animals or dissected organs, making global molecular phenotyping a reality for animals. Tandem mass spectrometry operated in data-independent acquisition can be used to compare the response of sentinels to various environmental conditions. The current low number of well-annotated animal or plant genomes, the high diversity of genetic backgrounds of each species, and the paucity of knowledge about protein functions for most of the relevant sentinels pose huge challenges for data interpretation. As a result, ecology and ecotoxicology today constitute an exceptional field for proteomics.

Evaluation of plant protection efficacy in field conditions and side effects of Lysobacter capsici AZ78, a biocontrol agent of Plasmopara viticola

ABSTRACT Downy mildew, caused by Plasmopara viticola, is a highly significant grapevine disease causing severe yield losses and affecting wine quality. Disease control mainly relies on use of copper and synthetic fungicides. Today there is a growing interest in developing microbial biopesticides for downy mildew control in integrated and organic viticulture. Here, the bacterial biocontrol agent Lysobacter capsici AZ78 (AZ78) was tested following a holistic approach including in planta efficacy test (large-scale trial) combined with bioassays aiming to assess potential adverse effects on non-target organisms: beneficial arthropods (Aphidius rhopalosiphi and Typhlodromus pyri), the crustacean Daphnia magna, the algae Selenastrum capricornutum and soil organisms [earthworm Eisenia (Ei.) fetida and worm Enchytraeid (En.) albidus]. Moreover, side effects of AZ78 treatments on grape quality and wine-making process were assessed. The results showed that AZ78 was equally effective to reference fungicide (Kocide®2000) in controlling downy mildew on leaves and bunches. AZ78 had no significant effect on the mortality of the indicator arthropod species at a Limit test Rate, which was equal to four times the recommended field rate. Additionally, no significant effects (mortality, growth rate or reproduction) were observed on D. magna, Ei. albidus, En. fetida and S. capricornutum, indicating AZ78 low toxicity on non-target organisms. Likewise, the yeast population dynamics in fermenting musts was not affected, without compromising the fermentation processes and wine quality. Overall, this study provides new insights into the characterisation of the biocontrol agent AZ78 showing that this bacterium has the potential to be further developed as a commercial biopesticide.

Ecological Impact of End-of-Life-Tire (ELT)-Derived Rubbers: Acute and Chronic Effects at Organism and Population Levels.

Considering the large amount of tires that reach the end of life every year, the aim of this study was the evaluation of both acute and chronic effects of end-of-life-tire (ELT)-derived rubber granules (ELT-dg) and powder (ELT-dp) on a freshwater trophic chain represented by the green alga Pseudokirchneriella subcapitata, the crustacean Daphnia magna and the teleost Danio rerio (zebrafish). Adverse effects were evaluated at the organism and population levels through the classical ecotoxicological tests. Acute tests on D. magna and D. rerio revealed a 50% effect concentration (EC50) > 100.0 mg/L for both ELT-dg and ELT-dp. Chronic exposures had a lowest observed effect concentration (LOEC) of 100.0 mg/L for both ELT-dg and ELT-dp on P. subcapitata grow rate and yield. LOEC decreased in the other model organisms, with a value of 9.8 mg/L for D. magna, referring to the number of living offspring, exposed to ELT-dg suspension. Similarly, in D. rerio, the main results highlighted a LOEC of 10.0 mg/L regarding the survival and juvenile weight parameters for ELT-dg and a LOEC of 10.0 mg/L concerning the survival and abnormal behavior in specimens exposed to ELT-dp. Tested materials exhibited a threshold of toxicity of 9.8 mg/L, probably a non-environmental concentration, although further investigations are needed to clarify the potential ecological impact of these emerging contaminants.

Antiparasitic potential of agrochemical fungicides on a non-target aquatic model (Daphnia × Metschnikowia host-parasite system)

Pesticides are a major anthropogenic threat to the biodiversity of freshwater ecosystems, having the potential to affect non-target aquatic organisms and disrupt the processes in which they intervene. Important knowledge gaps have been recognised concerning the ecological effects of synthetic fungicides on non-target symbiotic aquatic fungi and the ecological processes where they intervene. The goal of this work was to assess the influence of three commonly used fungicides (myclobutanil, metalaxyl and cymoxanil), which differ in their mode of action, on a host (the crustacean Daphnia magna) × parasite (the yeast Metschnikowia bicuspidata) experimental model. Using a set of life history experiments, we evaluated the effect of each fungicide on the outcome of this relationship (disease) and on the fitness of both host and parasite. Contrasting results were observed: (i) cymoxanil and metalaxyl were overall innocuous to host and parasite at the tested concentrations, although host reproduction was occasionally reduced in the simultaneous presence of parasite and fungicide; (ii) on the contrary, myclobutanil displayed a clear antifungal effect, decreasing parasite prevalence and alleviating infection signs in the hosts. This antiparasitic effect of myclobutanil was further investigated with a follow-up experiment that manipulated the timing of application of the fungicide, to understand which stage of parasite development was most susceptible: while myclobutanil did not interfere in the early stages of infection, its antifungal activity was clearly observable at a later stage of the disease (by impairing the production of transmission stages of the parasite). More research is needed to understand the broader consequences of this parasite-clearance effect, especially in face of increasing evidence that parasites are ecologically more important than their cryptic nature might suggest.

Transcriptional responses of Daphnia magna exposed to Akaki river water

Pollution of the aquatic environment is a global problem, with industrial waste, farming effluents, sewage, and wastewater as the main contributors. Many pollutants are biologically active at low concentrations, resulting in sublethal effects, which makes it a highly complex situation and difficult to assess. In many places, such as the Akaki river in Ethiopia, the pollution situation has resulted in streams with minimal presence of invertebrates or vertebrates. As it is difficult to perform a complete chemical analysis of the waters, the present study focused on using gene expression analysis as a biological end point to determine the effects of Akaki river contaminants. The present study was conducted using the small planktonic crustacean Daphnia magna with toxicogenomic molecular markers. Daphnia magna neonates were exposed to Akaki water samples collected from two different sites on the river and analyzed for mortality and expression of genes involved in different biological pathways. Despite the poor quality of Akaki river water, 48 h acute toxicity tests showed no mortality. Interestingly, analysis of sublethal toxicogenomic responses showed that exposure to Akaki water altered the expression of 25 out of 37 genes involved in metal regulation, immune response, oxidative stress, respiration, reproduction, and development. The toxicogenomic data gives insight into the mechanisms involved in causing potential adverse effects to aquatic biota harboring the Akaki river system.

Attenuation of toxicity and occurrence of degradation products of the fungicide tebuconazole after combined vacuum UV and UVC treatment of drinking water

Antifungal azoles are the most frequently used fungicides worldwide and occur as active ingredients in many antifungal pharmaceuticals, biocides, and pesticides. Azole fungicides are frequent environmental contaminants and can affect the quality of surface waters, groundwater, and drinking water. This study examined the potential of combined vacuum UV (185 nm) and UVC (254 nm) irradiation (VUV/UVC) of the azole fungicide tebuconazole and the transformation product 1,2,4-trizole on degradation and changes in ecotoxicity. In vivo ecotoxicity was examined before and after UV treatment using bioassays with test organisms from different trophic levels to integrate changes in biological effect of the parent compound and the degradation products. The test battery included the luminescent bacterium Aliivibrio fischeri, the Gram-positive bacterium Bacillus subtilis, the fungus Fusarium graminearum, the green microalga Raphidocelis subcapitata, and the crustacean Daphnia magna. The combined VUV/UVC treatment of tebuconazole in drinking water efficiently degraded the parent compound at the µg/L-mg/L level and resulted in transformation products with lower toxicity than the parent compound. A direct positive correlation was observed between the applied UV dose (fluence, J/cm2), the disappearance of tebuconazole, and the decrease in ecotoxicity. The combined VUV/UVC process does not require addition of supplementary oxidants or catalysts and our study suggests that VUV/UVC-mediated photolysis of azole fungicides in water can decrease the overall toxicity and represent a potentially environmentally friendly treatment method.

Demographic history shapes genomic variation in an intracellular parasite with a wide geographical distribution.

Analysing variation in a species' genomic diversity can provide insights into its historical demography, biogeography and population structure, and thus its ecology and evolution. Although such studies are rarely undertaken for parasites, they can be highly revealing because of the parasite's co-evolutionary relationships with hosts. Modes of reproduction and transmission are thought to be strong determinants of genomic diversity for parasites and vary widely among microsporidia (fungal-related intracellular parasites), which are known to have high intraspecific genetic diversity and interspecific variation in genome architecture. Here we explore genomic variation in the microsporidium Hamiltosporidium, a parasite of the freshwater crustacean Daphnia magna, looking especially at which factors contribute to nucleotide variation. Genomic samples from 18 Eurasian populations and a new, long-read-based reference genome were used to determine the roles that reproduction mode, transmission mode and geography play in determining population structure and demographic history. We demonstrate two main Hamiltosporidium tvaerminnensis lineages and a pattern of isolation-by-distance, but note an absence of congruence between these two parasite lineages and the two Eurasian host lineages. We suggest a comparatively recent parasite spread through Northern Eurasian host populations after a change from vertical to mixed-mode transmission and the loss of sexual reproduction. While gaining knowledge about the ecology and evolution of this focal parasite, we also identify common features that shape variation in genomic diversity for many parasites, such as distinct modes of reproduction and the intertwining of host-parasite demographies.

Intelligent high-throughput intervention testing platform in Daphnia.

We present a novel platform for testing the effects of interventions on the life‐ and healthspan of a short‐lived freshwater organism with complex behavior and physiology—the planktonic crustacean Daphnia magna. Within this platform, dozens of complex behavioral features of both routine motion and response to stimuli are continuously quantified over large synchronized cohorts via an automated phenotyping pipeline. We build predictive machine‐learning models calibrated using chronological age and extrapolate onto phenotypic age. We further apply the model to estimate the phenotypic age under pharmacological perturbation. Our platform provides a scalable framework for drug screening and characterization in both life‐long and instant assays as illustrated using a long‐term dose‐response profile of metformin and a short‐term assay of well‐studied substances such as caffeine and alcohol.

Development of transgenic Daphnia magna for visualizing homology-directed repair of DNA

In the crustacean Daphnia magna, studying homology-directed repair (HDR) is important to understand genome maintenance during parthenogenesis, effects of environmental toxicants on the genome, and improvement of HDR-mediated genome editing. Here we developed a transgenic D. magna that expresses green fluorescence protein (GFP) upon HDR occurrence. We utilized the previously established reporter plasmid named DR-GFP that has a mutated eGFP gene (SceGFP) and the tandemly located donor GFP gene fragment (iGFP). Upon double-strand break (DSB) introduction on SceGFP, the iGFP gene fragment acts as the HDR template and restores functional eGFP expression. We customized this reporter plasmid to allow bicistronic expression of the mCherry gene under the control of the D. magna EF1α-1 promoter/enhancer. By CRISPR/Cas-mediated knock-in of this plasmid via non-homologous joining, we generated the transgenic D. magna that expresses mCherry ubiquitously, suggesting that the DR-GFP reporter gene is expressed in most cells. Introducing DSB on the SceGFP resulted in eGFP expression and this HDR event could be detected by fluorescence, genomic PCR, and quantitative reverse-transcription PCR, suggesting this line could be used for evaluating HDR. The established reporter line might expand our understanding of the HDR mechanism and also improve the HDR-based gene-editing system in this species.

Toxicity of the antiparasitic lipopeptide biosurfactant SPH6 to green algae, cyanobacteria, crustaceans and zebrafish.

A lipopeptide with biosurfactant properties produced by the bacterium Pseudomonas H6 (SPH6) has antiparasitic effects and may serve as an alternative to chemotherapeutants against aquatic pathogens in aquaculture. We have elucidated its ecotoxicological potential by short-term standardized tests, including a growth rate inhibition test with algae (Raphidocelis subcapitata), a lethality test on the cyanobacteria Phormidium autumnale, a lethality test using crustaceans (Daphnia magna), a fish embryo acute toxicity test and a fish acute toxicity test using zebrafish (Danio rerio). The decrease of the biosurfactant concentration in zebrafish test water during 24 h was measured. The toxicity for crustaceans was highest (LC50 = 20 mg/L), followed by the test with the zebrafish embryo (LC50 = 27 mg/L). The juvenile zebrafish fish (complete mortality occurred between 40 and 80 mg/L), the cyanobacteria (LC50 = 80 mg/L) and the green algae (EC50 = 170 mg/L) showed higher tolerance. The determination of SPH6 concentrations in fish tank (up to 50% elimination over 24 h) suggested that the compound may become adsorbed to tank walls, absorbed by fish or degraded. Further studies should determine its impact under different environmental settings (e.g. temperature) relevant for different branches of the aquaculture sector.

Toxicity of micro and nano tire particles and leachate for model freshwater organisms

Environmental sampling has documented a diversity of microplastics, including high levels of black rubber— generally identified as tire debris. Though organisms have been shown to ingest tire particles (TPs), past research focused on toxicity of leachate alone, overlooking potential effects of particles. To address these gaps, we assessed the toxicity of micro (1–20 µm) and nano (<1 µm) TPs for two model organisms, embryonic Zebrafish Danio rerio and the crustacean Daphnia magna. To assess effects on development, Zebrafish embryos were exposed to concentrations of TPs or leachate ranging from 0 to 3.0 × 109 particles/ml and 0–100% respectively (n = 4). Greater mortality and sublethal malformations were observed following nano TP and leachate exposures as compared to micro TPs. Unique abnormalities between the exposures indicates that there is both chemical and particle-specific toxicity. We also observed D. magna mortality following a 48 h exposure of neonate to TPs or leachate, ranging from 0 to 3.3 × 109 particles/ml and 0–100% respectively (n = 3). Though, particle-enhancement of toxicity was observed for both Zebrafish and D. magna, overall sensitivity to TPs differed. It is important to identify differential toxicities across species to achieve an understanding of the environmental impacts of TPs and the chemicals they leach.

Environmental hazard of polypropylene microplastics from disposable medical masks: acute toxicity towards Daphnia magna and current knowledge on other polypropylene microplastics

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.