[en] Microplastics (MPs) are increasingly associated with physiological disruptions in aquatic organisms, yet the biological responses to environmentally sourced particles remain underexplored. This study investigated the reproductive toxicity of environmentally derived MPs collected from the Ikopa River (Antananarivo, Madagascar) in Danio rerio. Zebrafish were chronically exposed to cryomilled riverine MPs (1.2–50 µm) at concentrations of 100 and 1000 µg/L for 66 days, with daily reproductive assessments conducted over the final 21 days in accordance with OECD Test Guideline 229. Microplastic accumulation in gonadal tissue was assessed, along with subcellular responses via enzymatic assays in gonads and proteomic profiling in liver samples. Reproductive toxicity was evaluated through gonadal histology, fecundity, and fertility rates. MPs accumulated in gonads in a sex- and concentration dependent manner, with the highest levels in males exposed to 1000 µg/L (177.88 ± 102.65 particles/mg tissue, mean ± SD, n = 4). Despite MPs accumulation, no histopathological lesions were observed. However, significant oxidative stress and energy metabolism disruptions were identified in the liver, suggesting hepatic dysfunction as a potential driver of reproductive impairments. Furthermore, six polychlorinated biphenyl (PCB) congeners ranging from dozens to hundreds of ng/g MPs, and seven polybrominated diphenyl ether (PBDE) congeners in the range of a few ng/g MPs were detected on MPs surfaces, which may exacerbate toxicity via apoptosis inhibition. These findings provide novel mechanistic insights into how environmentally relevant MPs impair reproductive function in fish. The results underscore the necessity of incorporating environmental microplastics into toxicity testing frameworks to ensure accurate ecological risk assessment.
Disciplines :
Environmental sciences & ecology
Author, co-author :
Rabezanahary, Andry Ny Aina; University of Namur, Research Unit in Environmental and Evolutionary Biology (URBE), Institute of Life, Earth & Environment, Namur, Belgium ; Centre National de Recherches sur L'Environnement (CNRE), Rue Rasamimanana Fiadanana, Madagascar ; Plateforme Proteogen, Université de Caen Normandie, Esplanade de la paix, Caen, France
Ducarme, Baptiste; University of Namur, Research Unit in Environmental and Evolutionary Biology (URBE), Institute of Life, Earth & Environment, Namur, Belgium
Rajoelisoa, Andriamalala; Centre National de Recherches sur L'Environnement (CNRE), Rue Rasamimanana Fiadanana, Madagascar
Mong, Yves Jean Michel; Centre National de Recherches sur L'Environnement (CNRE), Rue Rasamimanana Fiadanana, Madagascar
Azari, Aala; Environment and Health, Department of Public Health and Primary Care, Leuven, Belgium
Ghosh, Manosij; Environment and Health, Department of Public Health and Primary Care, Leuven, Belgium
Hoet, Peter H.M.; Environment and Health, Department of Public Health and Primary Care, Leuven, Belgium
Muller, Hugo ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Eppe, Gauthier ; Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Bernay, Benoît; Plateforme Proteogen, Université de Caen Normandie, Esplanade de la paix, Caen, France
Randrianarivo, Ranjàna Hanitra; Department of Fundamental and Applied Biochemistry, Faculty of Science, University of Antananarivo, Madagascar
Kestemont, Patrick; University of Namur, Research Unit in Environmental and Evolutionary Biology (URBE), Institute of Life, Earth & Environment, Namur, Belgium
Missawi, Omayma; University of Namur, Research Unit in Environmental and Evolutionary Biology (URBE), Institute of Life, Earth & Environment, Namur, Belgium
Cornet, Valérie; University of Namur, Research Unit in Environmental and Evolutionary Biology (URBE), Institute of Life, Earth & Environment, Namur, Belgium
ARES - Academy of Research and Higher Education F.R.S.-FNRS - Fund for Scientific Research
Funding text :
This work was supported by the Scientific Research Fund (FNRS, Belgium) ( PDR N\u00B040003703 ) and the Academy of Research and Higher Education (ARES).
Baekelandt, S., Leroux, N., Burattin, L., Gérard, C., Delierneux, C., Robert, J.B., Cornet, V., Kestemont, P., Estetrol has a lower impact than 17α-ethinylestradiol on the reproductive capacity of zebrafish (Danio rerio). Aquat. Toxicol., 259, 2023, 10.1016/j.aquatox.2023.106505.
Bhagat, J., Zang, L., Nishimura, N., Shimada, Y., Zebrafish: an emerging model to study microplastic and nanoplastic toxicity. Sci. Total Environ., 728, 2020, 138707, 10.1016/j.scitotenv.2020.138707.
Biginagwa, F.J., Mayoma, B.S., Shashoua, Y., Syberg, K., Khan, F.R., First evidence of microplastics in the African Great Lakes: recovery from Lake Victoria Nile perch and Nile tilapia. J. Great Lakes Res. 42 (2016), 146–149, 10.1016/j.jglr.2015.10.012.
Bohnenberger, S., Wagner, B., Schmitz, H.J., Schrenk, D., Inhibition of apoptosis in rat hepatocytes treated with “non-dioxinlike” PCBs. Fresenius Environ. Bull. 12 (2003), 232–239.
Bucci, K., Tulio, M., Rochman, C.M., What is known and unknown about the effects of plastic pollution: a meta-analysis and systematic review. Ecol. Appl., 30, 2020, 10.1002/eap.2044.
Burns, E.E., Boxall, A.B.A., Microplastics in the aquatic environment: evidence for or against adverse impacts and major knowledge gaps. Environ. Toxicol. Chem. 37 (2018), 2776–2796, 10.1002/etc.4268.
Chen, Q., Gundlach, M., Yang, S., Jiang, J., Velki, M., Yin, D., Hollert, H., Quantitative investigation of the mechanisms of microplastics and nanoplastics toward zebrafish larvae locomotor activity. Sci. Total Environ. 584–585 (2017), 1022–1031, 10.1016/j.scitotenv.2017.01.156.
Clough, M.E., Ochoa Rivera, E., Parham, R.L., Ault, A.P., Zimmerman, P.M., McNeil, A.J., Tewari, A., Enhancing confidence in microplastic spectral identification via conformal prediction. Environ. Sci. Technol. 58 (2024), 21740–21749, 10.1021/acs.est.4c05167.
Cole, M., Lindeque, P., Halsband, C., Galloway, T.S., Microplastics as contaminants in the marine environment: a review. Mar. Pollut. Bull. 62 (2011), 2588–2597, 10.1016/j.marpolbul.2011.09.025.
Cormier, B., Cachot, J., Blanc, M., Cabar, M., Christelle, C., Dubocq, F., Le Bihanic, F., Morin, B., Zapata, S., Bégout, M.-L., Cousin, X., Environmental microplastics disrupt swimming activity in acute exposure in D. Rerio larvae and reduce growth and reproduction success in chronic exposure in D. Rerio and O. Melastigma. SSRN Electron. J., 308, 2022, 10.2139/ssrn.4059781.
Cormier, B., Gambardella, C., Tato, T., Perdriat, Q., Costa, E., Veclin, C., Le Bihanic, F., Grassl, B., Dubocq, F., Kärrman, A., Van Arkel, K., Lemoine, S., Lagarde, F., Morin, B., Garaventa, F., Faimali, M., Cousin, X., Bégout, M.L., Beiras, R., Cachot, J., Chemicals sorbed to environmental microplastics are toxic to early life stages of aquatic organisms. Ecotoxicol. Environ. Saf., 208, 2021, 10.1016/j.ecoenv.2020.111665.
Cui, X., Yang, T., Li, Z., Nowack, B., Meta-analysis of the hazards of microplastics in freshwaters using species sensitivity distributions. J. Hazard. Mater., 463, 2024, 132919, 10.1016/j.jhazmat.2023.132919.
De Frond, H.L., van Sebille, E., Parnis, J.M., Diamond, M.L., Mallos, N., Kingsbury, T., Rochman, C.M., Estimating the mass of chemicals associated with ocean plastic pollution to inform mitigation efforts. Integr. Environ. Assess. Manag. 15 (2019), 596–606, 10.1002/ieam.4147.
Ding, P., Xiang, C., Yao, Q., Li, X., Zhang, J., Yin, R., Zhang, L., Li, A.J., Hu, G., Aged polystyrene microplastics exposure affects apoptosis via inducing mitochondrial dysfunction and oxidative stress in early life of zebrafish. J. Environ. Manage., 367, 2024, 121995, 10.1016/j.jenvman.2024.121995.
Faker, M., Razavi Aghjeh, M.K., Ghaffari, M., Seyyedi, S.A., Rheology, morphology and mechanical properties of polyethylene/ethylene vinyl acetate copolymer (PE/EVA) blends. Eur. Polym. J. 44 (2008), 1834–1842, 10.1016/j.eurpolymj.2008.04.002.
Frias, J.P.G.L., Nash, R., Microplastics: finding a consensus on the definition. Mar. Pollut. Bull. 138 (2019), 145–147, 10.1016/j.marpolbul.2018.11.022.
Gao, Y., Fan, K.Y., Wei, J.X., Zeng, Y.Y., Liu, Q.F., Wang, C., Effects of microplastic concentrations on the survivability and reproduction of Moina Macrocopa. Appl. Ecol. Environ. Res. 21 (2023), 4155–4163, 10.15666/aeer/2105_41554163.
Gardon, T., Paul-pont, I., Le, G., Soyez, C., Lagarde, F., Cryogrinding and sieving techniques as challenges towards producing controlled size range microplastics for relevant ecotoxicological tests ☆. Environ. Pollut., 315, 2022, 120383, 10.1016/j.envpol.2022.120383.
Harikrishnan, T., Paramasivam, P., Sankar, A., Sakthivel, M., Sanniyasi, E., Raman, T., Thangavelu, M., Singaram, G., Muthusamy, G., Weathered polyethylene microplastics induced immunomodulation in zebrafish. Environ. Toxicol. Pharmacol., 109, 2024, 104478, 10.1016/j.etap.2024.104478.
Hou, T., Yu, J., Li, C., Wang, Z., Liu, H., Immunotoxicity of microplastics and polychlorinated biphenyls alone or in combination to Crassostrea gigas. Mar. Pollut. Bull., 200, 2024, 116161, 10.1016/j.marpolbul.2024.116161.
Hu, M., Palić, D., Micro- and nano-plastics activation of oxidative and inflammatory adverse outcome pathways. Redox Biol, 37, 2020, 10.1016/j.redox.2020.101620.
Hutton, S.J., Kashiwabara, L., Anderson, E., Siddiqui, S., Harper, B., Harper, S., Brander, S.M., Behavioral and molecular effects of micro and nanoplastics across three plastic types in fish: weathered microfibers induce a similar response to nanosized particles. Front. Toxicol. 6 (2024), 1–14, 10.3389/ftox.2024.1490223.
Jang, M., Shim, W.J., Han, G.M., Cho, Y., Moon, Y., Hong, S.H., Relative importance of aqueous leachate versus particle ingestion as uptake routes for microplastic additives (hexabromocyclododecane) to mussels. Environ. Pollut., 270, 2021, 116272, 10.1016/j.envpol.2020.116272.
Jia, Z., Zhao, C., Wang, M., Zhao, X., Zhang, W., Han, T., Xia, Q., Han, Z., Lin, R., Li, X., Hepatotoxicity assessment of Rhizoma Paridis in adult zebrafish through proteomes and metabolome. Biomed. Pharmacother., 121, 2020, 109558, 10.1016/j.biopha.2019.109558.
Junaid, M., Siddiqui, J.A., Liu, S., Lan, R., Abbas, Z., Chen, G., Wang, J., Adverse multigeneration combined impacts of micro(nano)plastics and emerging pollutants in the aquatic environment. Sci. Total Environ., 882, 2023, 163679, 10.1016/j.scitotenv.2023.163679.
Kameda, Y., Yamada, N., Fujita, E., Source- and polymer-specific size distributions of fine microplastics in surface water in an urban river. Environ. Pollut., 284, 2021, 117516, 10.1016/j.envpol.2021.117516.
Kim, J.H., Yu, Y.Bin, Choi, J.H., Toxic effects on bioaccumulation, hematological parameters, oxidative stress, immune responses and neurotoxicity in fish exposed to microplastics: a review. J. Hazard. Mater., 413, 2021, 125423, 10.1016/j.jhazmat.2021.125423.
Li, C., Busquets, R., Campos, L.C., Assessment of microplastics in freshwater systems: a review. Sci. Total Environ., 707, 2020, 135578, 10.1016/j.scitotenv.2019.135578.
Li, Z., Chang, X., Hu, M., Fang, J.K.H., Sokolova, I.M., Huang, W., Xu, E.G., Wang, Y., Is microplastic an oxidative stressor? Evidence from a meta-analysis on bivalves. J. Hazard. Mater., 423, 2022, 127211, 10.1016/j.jhazmat.2021.127211.
Lu, Y., Zhang, Y., Deng, Y., Jiang, W., Zhao, Y., Geng, J., Ding, L., Ren, H., Uptake and accumulation of polystyrene microplastics in zebrafish (Danio rerio) and toxic effects in liver. Environ. Sci. Technol. 50 (2016), 4054–4060, 10.1021/acs.est.6b00183.
Mai, N.T.Q., Batjargal, U., Kim, W.S., Kim, J.H., Park, J.W., Kwak, I.S., Moon, B.S., Microplastic induces mitochondrial pathway mediated cellular apoptosis in mussel (Mytilus galloprovincialis) via inhibition of the AKT and ERK signaling pathway. Cell Death Discov. 9 (2023), 1–10, 10.1038/s41420-023-01740-3.
McColley, C.J., Nason, J.A., Harper, B.J., Harper, S.L., An assessment of methods used for the generation and characterization of cryomilled polystyrene micro- and nanoplastic particles. Microplast.Nanoplast, 3, 2023, 10.1186/s43591-023-00069-z.
Michailidou, K., Palisidou, C., Feidantsis, K., Ainali, N.M., Kastrinaki, G., Lambropoulou, D.A., Kyzas, G.Z., Bikiaris, D.N., Kaloyianni, M., Bobori, D.C., Impact of aged and virgin polyethylene microplastics on multi end-points effects of freshwater fish tissues. Sci. Total Environ., 948, 2024, 174704, 10.1016/j.scitotenv.2024.174704.
Missawi, O., Bousserrhine, N., Zitouni, N., Maisano, M., Boughattas, I., De Marco, G., Cappello, T., Belbekhouche, S., Guerrouache, M., Alphonse, V., Banni, M., Uptake, accumulation and associated cellular alterations of environmental samples of microplastics in the seaworm Hediste diversicolor. J. Hazard. Mater., 406, 2021, 124287, 10.1016/j.jhazmat.2020.124287.
Missawi, O., Jeddou, I.Ben, Venditti, M., Zitouni, N., Zaouali, M.A., Abdennebi, H.Ben, Messaoudi, I., Reiter, R.J., Minucci, S., Banni, M., Environmental microplastic accumulation exacerbates liver ischemia-reperfusion injury in rat: protective effects of melatonin. Sci. Total Environ., 860, 2023, 160155, 10.1016/j.scitotenv.2022.160155.
Mohamed Helal, Sara Ghanem, A.E.-S., Impact of PCBs, Furan and Dioxin on hepatocarcinogenesis. Persistent org. Pollut. - monit. Impact treat. Immunol., 11, 2022, 13, 10.5772/intechopen.101526.
Mohamed Nor, N.H., Koelmans, A.A., Transfer of PCBs from microplastics under simulated gut fluid conditions is biphasic and reversible. Environ. Sci. Technol. 53 (2019), 1874–1883, 10.1021/acs.est.8b05143.
Morozov, A.A., Yurchenko, V.V., Changes in the liver proteome of zebrafish (Danio rerio) exposed to glyphosate and aminomethylphosphonic acid in the presence of a humic substance. Pestic. Biochem. Physiol., 204, 2024, 106036, 10.1016/j.pestbp.2024.106036.
Muller, H.B., Scholl, G., Far, J., De Pauw, E., Eppe, G., Sliding windows in ion mobility (SWIM): a new approach to increase the resolving power in trapped ion mobility-mass spectrometry hyphenated with chromatography. Anal. Chem. 95 (2023), 17586–17594, 10.1021/acs.analchem.3c03039.
Nam, T.G., Lipid peroxidation and its toxicological implications. Toxicol. Res. 27 (2011), 1–6, 10.5487/TR.2011.27.1.001.
OECD, 2010. Testing and assessment No. 123 guidance document on the diagnosis of endocrine-related histopathology in fish gonads JT03284402.
OECD, 2009. Test No. 229: fish short term reproduction assay. Test No. 229 fish short term reprod. Assay. https://doi.org/10.1787/9789264076211-en.
Pannetier, P., Morin, B., Le Bihanic, F., Dubreil, L., Clérandeau, C., Chouvellon, F., Van Arkel, K., Danion, M., Cachot, J., Environmental samples of microplastics induce significant toxic effects in fish larvae. Environ. Int., 134, 2020, 105047, 10.1016/j.envint.2019.105047.
Patchaiyappan, A., Dowarah, K., Zaki Ahmed, S., Prabakaran, M., Jayakumar, S., Thirunavukkarasu, C., Devipriya, S.P., Prevalence and characteristics of microplastics present in the street dust collected from Chennai metropolitan city. India. Chemosphere, 269, 2021, 128757, 10.1016/j.chemosphere.2020.128757.
Phuong, N.N., Zalouk-Vergnoux, A., Poirier, L., Kamari, A., Châtel, A., Mouneyrac, C., Lagarde, F., Is there any consistency between the microplastics found in the field and those used in laboratory experiments?. Environ. Pollut. 211 (2016), 111–123, 10.1016/j.envpol.2015.12.035.
Pisoschi, A.M., Pop, A., The role of antioxidants in the chemistry of oxidative stress: a review. Eur. J. Med. Chem. 97 (2015), 55–74, 10.1016/j.ejmech.2015.04.040.
Qiang, L., Cheng, J., Exposure to polystyrene microplastics impairs gonads of zebrafish (Danio rerio). Chemosphere, 263, 2021, 128161, 10.1016/j.chemosphere.2020.128161.
Qiao, R., Deng, Y., Zhang, S., Wolosker, M.B., Zhu, Q., Ren, H., Zhang, Y., Accumulation of different shapes of microplastics initiates intestinal injury and gut microbiota dysbiosis in the gut of zebrafish. Chemosphere, 236, 2019, 124334, 10.1016/j.chemosphere.2019.07.065.
Quigley, E., Silva, A.L.P., Chelinho, S., Cunha, L., Briones, M.J., Sousa, J.P., Pristine and UV-aged polyethylene microplastics’ impact on gut microbiome and reproduction of earthworm Eisenia andrei. Eur. J. Soil Biol., 122, 2024, 10.1016/j.ejsobi.2024.103640.
Rabezanahary, A., Kestemont, P., Cornet, V., Benali, S., Laby, P., Hanitra, R., Yves, R., Michel, J., Jean, M., Raquez, M., Missawi, O., Unseen riverine risk: spatio temporal shifts of microplastic pollution and its bioavailability in freshwater fish within the Ikopa River urban system. Environ. Monit. Assess., 2024, 10.1007/s10661-024-13010-5.
Rozman, U., Kal, G., 2022. Seeking for a perfect (non-spherical) microplastic particle – The most comprehensive review on microplastic laboratory research 424. https://doi.org/10.1016/j.jhazmat.2021.127529.
Shruti, V.C., Kutralam-muniasamy, G., Trends in environmental analytical chemistry blanks and bias in microplastic research: implications for future quality assurance. Trends Environ. Anal. Chem., 38, 2023, e00203, 10.1016/j.teac.2023.e00203.
Singleman, C., Holtzman, N.G., Growth and maturation in the zebrafish, Danio Rerio: a staging tool for teaching and research. Zebrafish 11 (2014), 396–406, 10.1089/zeb.2014.0976.
Sokolova, I.M., Frederich, M., Bagwe, R., Lannig, G., Sukhotin, A.A., Energy homeostasis as an integrative tool for assessing limits of environmental stress tolerance in aquatic invertebrates. Mar. Environ. Res. 79 (2012), 1–15, 10.1016/j.marenvres.2012.04.003.
Tran, T.K.A., Yu, R.M.K., Islam, R., Nguyen, T.H.T., Bui, T.L.H., Kong, R.Y.C., O'Connor, W.A., Leusch, F.D.L., Andrew-Priestley, M., MacFarlane, G.R., The utility of vitellogenin as a biomarker of estrogenic endocrine disrupting chemicals in molluscs. Environ. Pollut. 248 (2019), 1067–1078, 10.1016/j.envpol.2019.02.056.
Ullah, Sana, Ahmad, S., Guo, X., Ullah, Saleem, Ullah, Sana, Nabi, G., Wanghe, K., A review of the endocrine disrupting effects of micro and nano plastic and their associated chemicals in mammals. Front. Endocrinol. 13 (2023), 1–17, 10.3389/fendo.2022.1084236.
Varshney, S., Hegstad-Pettersen, M.M., Siriyappagouder, P., Olsvik, P.A., Enhanced neurotoxic effect of PCB-153 when co-exposed with polystyrene nanoplastics in zebrafish larvae. Chemosphere, 355, 2024, 141783, 10.1016/j.chemosphere.2024.141783.
Vroom, R.J.E., Koelmans, A.A., Besseling, E., Halsband, C., Aging of microplastics promotes their ingestion by marine zooplankton. Environ. Pollut. 231 (2017), 987–996, 10.1016/j.envpol.2017.08.088.
Wang, J., Li, Y., Lu, L., Zheng, M., Zhang, X., Tian, H., Wang, W., Ru, S., Polystyrene microplastics cause tissue damages, sex-specific reproductive disruption and transgenerational effects in marine medaka (Oryzias melastigma). Environ. Pollut., 254, 2019, 113024, 10.1016/j.envpol.2019.113024.
Wang, J., Wu, F., Dong, S., Wang, Xusheng, Ai, S., Liu, Z., Wang, Xiaonan, Meta-analysis of the effects of microplastic on fish: insights into growth, survival, reproduction, oxidative stress, and gut microbiota diversity. Water Res, 267, 2024, 122493, 10.1016/j.watres.2024.122493.
Wang, Z.M., Wagner, J., Ghosal, S., Bedi, G., Wall, S., SEM/EDS and optical microscopy analyses of microplastics in ocean trawl and fish guts. Sci. Total Environ. 603–604 (2017), 616–626, 10.1016/j.scitotenv.2017.06.047.
Xia, X., Guo, W., Ma, X., Liang, N., Duan, X., Zhang, P., Zhang, Y., Chang, Z., Zhang, X., Reproductive toxicity and cross-generational effect of polyethylene microplastics in Paramisgurnus dabryanus. Chemosphere, 313, 2023, 137440, 10.1016/j.chemosphere.2022.137440.
Xiong, X., Tu, Y., Chen, X., Jiang, X., Shi, H., Wu, C., Elser, J.J., Ingestion and egestion of polyethylene microplastics by goldfish (Carassius auratus): influence of color and morphological features. Heliyon, 5, 2019, e03063, 10.1016/j.heliyon.2019.e03063.
Yi, J., Ma, Y., Ruan, J., You, S., Ma, J., Yu, H., Zhao, J., Zhang, K., Yang, Q., Jin, L., Zeng, G., Sun, D., The invisible threat: assessing the reproductive and transgenerational impacts of micro- and nanoplastics on fish. Environ. Int., 183, 2024, 108432, 10.1016/j.envint.2024.108432.
Zhou, N., Wang, Z., Yang, L., Zhou, W., Qin, Z., Zhang, H., Size-dependent toxicological effects of polystyrene microplastics in the shrimp Litopenaeus vannamei using a histomorphology, microbiome, and metabolic approach. Environ. Pollut., 316, 2023, 120635, 10.1016/j.envpol.2022.120635.
