key: cord-0941008-lhdw7flm authors: Nugnes, Roberta; Lavorgna, Margherita; Orlo, Elena; Russo, Chiara; Isidori, Marina title: Toxic impact of polystyrene microplastic particles in freshwater organisms date: 2022-03-18 journal: Chemosphere DOI: 10.1016/j.chemosphere.2022.134373 sha: 78964e30d9c9ec80e3c5d03be7125207f1df5980 doc_id: 941008 cord_uid: lhdw7flm The ongoing COVID-19 pandemic is leading to an increase of the global production of plastics since the use of personal protective equipment (PPEs, i.e. gloves, gowns, masks, packaging items), has become mandatory to prevent the spread of the virus. Plastic breaks down into micro/nano particles due to physical or chemical or biological actions into environment. Due to small dimensions, ubiquitous and persistent nature, the plastic particles represent a significant threat to ecosystems and can entry into food chains. Among the plastic polymers used for PPEs, polystyrene is less studied regarding its eco-geno-toxicity. This study aims to investigate acute, chronic and subchronic effects of the microplastic polystyrene beads (PS-MP, size 1.0 μm) on three freshwater species, the alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus, the crustacean Ceriodaphnia dubia and the benthic ostracod Heterocypris incongruens. Furthermore, the potential genotoxicity and the ROS production due to the PS-MP were also determined in C. dubia. Results revealed that the acute effects occurred at concentrations of PS-MP in the order of dozens of mg/L in B. calyciflorus and C. dubia and hundreds of mg/L in H. incongruens. Regarding long-term toxicity, increasing chronic effects with EC50s in the order of units (C. dubia), hundreds (B. calyciflorus) and thousands (R. subcapitata) of μg/L were observed. Both for acute and chronic/sub chronic toxicity, daphnids were more sensitive to polystyrene than ostracods. Moreover, when C. dubia neonates were exposed to the PS-MP, alterations in genetic material as well as the production of ROS occurred, starting from concentrations in the order of units of μg/L, probably due to inflammatory responses. At last, the risk quotient (RQ) as a measure of risk posed by PS-MPs in freshwater environment, was calculated obtaining a value of 7.2, higher than the threshold value of 1. The microplastic particles of polystyrene have been found to be genotoxic to aquatic organisms 127 probably due to the production of reactive oxygen species (ROS), oxidative stress and interference 128 in DNA repair (Dong et al., 2019; Solomando et al., 2020) . 129 The present study aimed to broaden the knowledge on the acute, chronic and subchronic effects of risk posed by PS-MPs in freshwater environments. In order to establish the possible differences 138 between the nominal and actual concentrations, the concentration of PS-MP in the test solution was 139 also investigated. Monodispersed micro particles of polystyrene (PS-MP) with a solid content of 2% weight, 146 analytical standard size equal to 1.0 µm and particle specific gravity equal to 1.05 g/cm 3 , were 147 purchased from Sigma Aldrich (Milano, Italy) as aqueous suspension (21,000 mg/L) containing 148 approximate 3.24·10 10 particles/mL (PS, Product number: 72938). 149 Test solutions were freshly prepared by mixing the appropriate volumes of the aqueous suspension with one rotifer/well and 0.9 mL/well of test solutions (10 dilutions in moderately hard dilution 204 water, from 0.9 to 32·10 3 µg/L, geometric progression of 3.2, chosen after appropriate range finding immediately before the test and after 6 d. The growth inhibition was determined only when a mortality of less than 30% was found in the 223 acute assay. The comet assay was performed on cells coming from whole organisms. Neonates were less than 24 239 h old and coming from the second to the fifth brood, obtained by parthenogenesis. About 20 neonates were placed in glass beakers and exposed to polystyrene for 24 h, in darkness at Thus, in the present study, the effective concentrations were reported as nominal concentrations. referring to one third of the life span of the organisms. The results of EC50, EC20 and EC10 from three independent experiments are shown in Table 2 The sub-chronic effect was studied on the primary benthic consumer H. incongruens after 6 days of 359 exposure. As explained in the material and methods section, according to standard guidelines, ostracod length was only recorded when mortality was less than 30% and thus when survival was 361 greater than 70%. As can be seen in In addition, when comparing all NOEC and LOEC values from all chronic/sub-chronic tests, it is 377 always evident that among all organisms tested, C. dubia was the organism with the lowest NOEC 378 and LOEC values showing the highest significant no effect concentration and the lowest significant The genotoxicity evaluation was performed by the comet assay on the freshwater crustacean C. 387 dubia wholly exposed in vivo to polystyrene for 24 h. As reported in Fig.2 The gut content of C. dubia after exposure to different concentrations of PS was analysed using a 424 light microscope and was visually inspected and compared to the negative control ( Figure S1 ). In 425 Figure S1a the gut tract of non-exposed C. dubia appeared completely empty and the whole 470 Surely, as reported by Gorokhova (2015) and De Felice and colleagues (2019) of particular concern 471 is the exposure and the subsequent effects due to microparticles in zooplanktonic filter-feeder 472 species, which indiscriminately ingest them during their normal swimming and feeding activity. In fact, as reported by Ebert (2005) , daphnids have the ability to uptake and ingest small (1-70 µm Besseling and coauthors observed that when the green chlorococcal alga Scenedesmus obliquus was Table S1 . the AF depended on the number of trophic levels considered which, in the present study, were two 537 (alga, rotifers and crustaceans, producers and primary consumers) (Table S1 ). Therefore, 538 considering all the parameters just mentioned, the value of PS-MP RQ was equal to 7.2 absolutely 539 above the threshold value of 1, with a severe environmental concern for the freshwater ecosystem. In light of the above, there is no doubt that during the COVID-19 era, the request, the production 541 and the consumption of plastic materials, including polystyrene products have grown exponentially Microplastic 599 contamination in freshwater environments: a review, focusing on interactions with sediments and 600 benthic organisms Genotoxic evaluation of polystyrene microplastic Nanoplastic affects growth of S. 606 obliquus and reproduction of D. magna Nanoplastics Identification 610 in Complex Environmental Matrices: Strategies for Polystyrene and Polypropylene Microplastic ingestion by Daphnia magna and its enhancement 614 on algal growth Food chain transport of 618 nanoparticles affects behaviour and fat metabolism in fish Polystyrene 648 microplastic particles: In vitro pulmonary toxicity assessment Some parasites of Daphnia Presence of microplastics and nanoplastics in food, with particular focus on 656 seafood: panel on contamination in the food chain Technical Guidance Document on risk Assessment Guideline on the Environmental Risk Assesment of the Medicinal Products for 664 Risk assessment of microplastics in the ocean: modelling approach and 668 first conclusions Screening for microplastic particles in plankton samples: How to integrate 671 marine litter assessment into existing monitoring programs? oxidative stress in clinical samples: research applications in the cancer field The molecular effects of acrolein Ultraviolet B retards growth, 700 induces oxidative stress, and modulates DNA repair-related gene and heat shock protein gene 701 expression in the monogonont Analysis, occurrence, and degradation 705 of microplastics in the aqueous environment Microplastics in freshwaters and drinking water: Critical review and assessment of data quality Prediction and 713 assessment of ecogenotoxicity of anti-neoplastic drugs in binary mixtures Toxicity and genotoxicity 717 of the quaternary ammonium compound benzalkonium chloride (BAC) using Daphnia 718 magna and Ceriodaphnia dubia as model systems Bioaccumulation of polystyrene nanoplastics and 722 their effect on the toxicity of Au ions in zebrafish embroys Investigating 744 the toxicities of different functionalized polystyrene nanoplastics on Daphnia magna Polystyrene nanoplastic 747 exposure induces immobilization, reproduction, and stress defense in the freshwater cladoceran 748 751 Optimization of epitope processing enhances immunogenicity of multiepitope DNA 752 vaccines Levels and fate of perfluoroalkyl 755 substances in beached plastic pellets and sediments collected from Greece Accumulation of Polystyrene Microplastics in Zebrafish (Danio rerio) and Toxic Effects in Liver Current status of production and supply of Face Masks, Antiseptics and Toilet Paper The effects of microplastics on freshwater Hydra attenuate 766 feeding, morphology and reproduction OECD Guideline for testing of chemicals 201: Freshwater Alga and 770 Performance of a miniaturized algal bioassay in phytotoxicity screening Plastics in the time of COVID-19 pandemic: Protector or polluter Cytotoxicity, genotoxicity and ecotoxicity assay using human cell and 780 environmental species for the screening of the risk from pollutant exposure Eco-genotoxicity of six 784 anticancer drugs using comet assay in daphnids Plastic waste release caused by COVID-19 792 and its fate in the global ocean Occurrence 796 of microplastics in raw and treated drinking water Low doses of widely consumed cannabinoids (cannabidiol 801 and cannabidivarin) cause DNA damage and chromosomal aberrations in human-derived cells Toxicity of microplastics and natural particles in the freshwater dipteran Chironomus riparius: 806 Same same but different? Trace 810 analysis of polystyrene microplastics in natural waters Analysis of microplastic in water 814 by micro-Raman spectroscopy: release of plastic particles from different packaging into mineral 815 water Plastic pollution during COVID-19: Plastic waste 818 directives and its long-term impact on the environment Environmental 822 challenges induced by extensive use of face masks during COVID-19: A review and potential 823 solutions Relevance of nano-and microplastics for freshwater 847 ecosystems: A critical review Methods for measuring the acute toxicity of effluents and receiving waters to 851 freshwater and marine organisms Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to 855 Freshwater and Marine Organisms Do microplastic 858 contaminated seafood consumption pose a potential risk to human health? Rational use of personal protective equipment (PPE) for 862 coronavirus disease (COVID-19 The physical impacts of microplastics on 866 marine organisms: A review phenanthrenequinone, and phenanthrene to Daphnia magna: evidence for a reactive oxygen 872 mechanism