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Mixture toxicity of pyraclostrobine and metiram to the zebrafish (Danio rerio) and its potential mechanism

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Abstract

The interplay between pesticides plays a critical role in ecotoxicology since these chemicals rarely emerge as single substances but rather in mixtures with other chemicals. In the present work, we purposed to clarify the combined toxic impacts of pyraclostrobine (PYR) and metiram (MET) on the zebrafish by using numerous indicators. Results exhibited that the 4-day LC50 value of MET to fish embryos was 0.0025 mg a.i. L−1, which was lower compared with PYR (0.019 mg a.i. L−1). Combinations of PYR and MET presented a synergetic impact on fish embryos. Contents of POD, CYP450, and VTG were drastically increased in the plurality of the single and joint treatments relative to the baseline value. Three genes, including vtg1, crh, and il-8, related to the endocrine and immune systems, were also surprisingly up-regulated when fish were challenged by the individual and mixture pesticides compared with the baseline value. These results afforded valuable information on the latent toxicity mechanisms of co-exposure for PYR and MET in the early growth stage of fish. Moreover, our data also revealed that frequent application of these two pesticides might exert a potentially ecotoxicological hazard on aquatic ecosystems. Collectively, the present study provided valuable guidance for the risk evaluation of chemical combinations.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

References

  • Abe FR, Accoroni KAG, Gravato C, de Oliveira DP (2021) Early life stage assays in zebrafish. Methods Mol Biol 2240:77–92

    CAS  Google Scholar 

  • Aksakal FI, Ciltas A (2019) Impact of copper oxide nanoparticles (CuO NPs) exposure on embryo development and expression of genes related to the innate immune system of zebrafish (Danio rerio). Comp Biochem Physiol C Toxicol Pharmacol 223:78–87

    CAS  Google Scholar 

  • Bhagat J, Singh N, Nishimura N, Shimada Y (2021) A comprehensive review on environmental toxicity of azole compounds to fish. Chemosphere 262:128335

    CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    CAS  Google Scholar 

  • Campos-Sánchez JC, Esteban MÁ (2021) Review of inflammation in fish and value of the zebrafish model. J Fish Dis 44(2):123–139

    Google Scholar 

  • Çayır A, Coşkun M, Coşkun M (2016) Genotoxicity of commercial fungicide Cabrio Plus on human cell. Cytotechnology 68(5):1697–1704

    Google Scholar 

  • Chagas TQ, Freitas ÍN, Montalvão MF, Nobrega RH, Machado MRF, Charlie-Silva I, Araújo APDC, Guimarães ATB, Alvarez TGDS, Malafaia G (2021) Multiple endpoints of polylactic acid biomicroplastic toxicity in adult zebrafish (Danio rerio). Chemosphere 277:130279

    CAS  Google Scholar 

  • Chen J, Zheng L, Tian L, Wang N, Lei L, Wang Y, Dong Q, Huang C, Yang D (2018) Chronic PFOS exposure disrupts thyroid structure and function in zebrafish. Bull Environ Contam Toxicol 101(1):75–79

    CAS  Google Scholar 

  • Chi H (1997) Computer program for the probit analysis. National Chung Hsing University, Taichung

    Google Scholar 

  • Cong B, Liu C, Wang L, Chai Y (2020) The impact on antioxidant enzyme activity and related gene expression following adult Zebrafish (Danio rerio) exposure to dimethyl phthalate. Animals (basel) 10(4):717

    Google Scholar 

  • Covert SA, Shoda ME, Stackpoole SM, Stone WW (2020) Pesticide mixtures show potential toxicity to aquatic life in US streams, water years 2013–2017. Sci Total Environ 745:141285

    CAS  Google Scholar 

  • de Lima D, Roque GM, de Almeida EA (2013) In vitro and in vivo inhibition of acetylcholinesterase and carboxylesterase by metals in zebrafish (Danio rerio). Mar Environ Res 91:45–51

    Google Scholar 

  • Di Paolo C, Groh KJ, Zennegg M, Vermeirssen EL, Murk AJ, Eggen RI, Hollert H, Werner I, Schirmer K (2015) Early life exposure to PCB126 results in delayed mortality and growth impairment in the zebrafish larvae. Aquat Toxicol 69:168–178

    Google Scholar 

  • El-Nahhal Y (2018) Toxicity of some aquatic pollutants to fish. Environ Monit Assess 190(8):449

    Google Scholar 

  • Félix LM, Luzio A, Antunes L, Coimbra AM, Valentim AM (2021) Malformations and mortality in zebrafish early stages associated with elevated caspase activity after 24 h exposure to MS-222. Toxicol Appl Pharmacol 412:115385

    Google Scholar 

  • Gaaied S, Oliveira M, Le Bihanic F, Cachot J, Banni M (2019) Gene expression patterns and related enzymatic activities of detoxification and oxidative stress systems in zebrafish larvae exposed to the 2,4-dichlorophenoxyacetic acid herbicide. Chemosphere 224:289–297

    CAS  Google Scholar 

  • Guo D, Liu W, Yao T, Ma M, Wang Q, Qiu J, Qian Y (2021) Combined endocrine disruptive toxicity of malathion and cypermethrin to gene transcription and hormones of the HPG axis of male zebrafish (Danio rerio). Chemosphere 267:128864

    CAS  Google Scholar 

  • Güvercin S, Erat M, Sakiroğlu H (2008) Determination of some kinetic and characteristic properties of glutathione S-transferase from bovine erythrocytes. Protein Pept Lett 15(1):6–12

    Google Scholar 

  • Heck DE, Shakarjian M, Kim HD, Laskin JD, Vetrano AM (2010) Mechanisms of oxidant generation by catalase. Ann N Y Acad Sci 1203:120–125

    CAS  Google Scholar 

  • Hemalatha D, Rangasamy B, Nataraj B, Maharajan K, Narayanasamy A, Ramesh M (2020) Transcriptional, biochemical and histological alterations in adult zebrafish (Danio rerio) exposed to benzotriazole ultraviolet stabilizer-328. Sci Total Environ 739:139851

    CAS  Google Scholar 

  • ISO (1996) Water quality-Determination of the acute lethal toxicity of substances to a freshwater fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)]-Part 3: Flow-through method. ISO 7346–3

  • Jia K, Cheng B, Huang L, Xiao J, Bai Z, Liao X, Cao Z, Shen T, Zhang C, Hu C, Lu H (2020) Thiophanate-methyl induces severe hepatotoxicity in zebrafish. Chemosphere 248:125941

    CAS  Google Scholar 

  • Kim DJ, Seok SH, Baek MW, Lee HY, Na YR, Park SH, Lee HK, Dutta NK, Kawakami K, Park JH (2009) Benomyl induction of brain aromatase and toxic effects in the zebrafish embryo. J Appl Toxicol 29(4):289–294

    CAS  Google Scholar 

  • Klittich CJR, Wang NX, Zhang Y, Rowland LB (2020) A revised model of fungicide translaminar activity. Pestic Biochem Physiol 167:104597

    CAS  Google Scholar 

  • Lee S, Eghan K, Lee J, Yoo D, Yoon S, Kim WK (2020) Zebrafish embryonic exposure to BPAP and its relatively weak thyroid hormone-disrupting effects. Toxics 8(4):103

    CAS  Google Scholar 

  • Lewandowski Ł, Kepinska M, Milnerowicz H (2018) Inhibition of copper-zinc superoxide dismutase activity by selected environmental xenobiotics. Environ Toxicol Pharmacol 58:105–113

    CAS  Google Scholar 

  • Li S, Jiang Y, Sun Q, Coffin S, Chen L, Qiao K, Gui W, Zhu G (2020) Tebuconazole induced oxidative stress related hepatotoxicity in adult and larval zebrafish (Danio rerio). Chemosphere 241:125129

    CAS  Google Scholar 

  • Li XY, Qin YJ, Wang Y, Huang T, Zhao YH, Wang XH, Martyniuk CJ, Yan B (2021) Relative comparison of strobilurin fungicides at environmental levels: Focus on mitochondrial function and larval activity in early staged zebrafish (Danio rerio). Toxicology 452:152706

    CAS  Google Scholar 

  • Liang YQ, Xu W, Liang X, Jing Z, Pan CG, Tian F (2020) The synthetic progestin norethindrone causes thyroid endocrine disruption in adult zebrafish. Comp Biochem Physiol C Toxicol Pharmacol 236:108819

    CAS  Google Scholar 

  • Lin CH, Chou PH, Chen PJ (2014) Two azole fungicides (carcinogenic triadimefon and non-carcinogenic myclobutanil) exhibit different hepatic cytochrome P450 activities in medaka fish. J Hazard Mater 277:150–158

    CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT Method. Methods 25:402–408

    CAS  Google Scholar 

  • Loerracher AK, Braunbeck T (2020) Inducibility of cytochrome P450-mediated 7-methoxycoumarin-O-demethylase activity in zebrafish (Danio rerio) embryos. Aquat Toxicol 225:105540

    CAS  Google Scholar 

  • Luo Y, Chen H, Li D, Zhan M, Hou L, Dong W, Luo Y, Xie L (2020) The effects of norethindrone on the ontogeny of gene expression along the hypothalamic-pituitary-adrenal and hypothalamic-pituitary-gonadal axes in zebrafish (Danio rerio). Sci Total Environ 747:141554

    CAS  Google Scholar 

  • Ma X, Li H, Xiong J, Mehler WT, You J (2019) Developmental toxicity of a neonicotinoid insecticide, acetamiprid to zebrafish embryos. J Agric Food Chem 67(9):2429–2436

    CAS  Google Scholar 

  • Majumder R, Kaviraj A (2019) Acute and sublethal effects of organophosphate insecticide chlorpyrifos on freshwater fish Oreochromis niloticus. Drug Chem Toxicol 42(5):487–495

    CAS  Google Scholar 

  • Manikandan P, Nagini S (2018) Cytochrome P450 structure, function and clinical significance: A Review. Curr Drug Targets 19(1):38–54

    CAS  Google Scholar 

  • Mansano AS, Moreira RA, Dornfeld HC, Freitas EC, Vieira EM, Daam MA, Rocha O, Seleghim MHR (2020) Individual and mixture toxicity of carbofuran and diuron to the protozoan Paramecium caudatum and the cladoceran Ceriodaphnia silvestrii. Ecotoxicol Environ Saf 201:110829

    CAS  Google Scholar 

  • Mao L, Jia W, Zhang L, Zhang Y, Zhu L, Sial MU, Jiang H (2020) Embryonic development and oxidative stress effects in the larvae and adult fish livers of zebrafish (Danio rerio) exposed to the strobilurin fungicides, kresoxim-methyl and pyraclostrobin. Sci Total Environ 729:139031

    CAS  Google Scholar 

  • Marking LL (1985) Toxicity of chemical mixtures. In: Rand G, Petrocelli QS (eds) Fundamentals of aquatic toxicology. Hemisphere Publishing Corporation, Washington DC, pp 164–176

    Google Scholar 

  • Martínez R, Tu W, Eng T, Allaire-Leung M, Piña B, Navarro-Martín L, Mennigen JA (2020) Acute and long-term metabolic consequences of early developmental Bisphenol A exposure in zebrafish (Danio rerio). Chemosphere 256:127080

    Google Scholar 

  • Novoa B, Figueras A (2012) Zebrafish: model for the study of inflammation and the innate immune response to infectious diseases. Adv Exp Med Biol 946:253–275

    CAS  Google Scholar 

  • Ons L, Bylemans D, Thevissen K, Cammune BPA (2020) Combining biocontrol agents with chemical fungicides for integrated plant fungal disease control. Microorganisms 8(12):1930

    CAS  Google Scholar 

  • Park CG, Sung B, Ryu CS, Kim YJ (2020) Mono-(2-ethylhexyl) phthalate induces oxidative stress and lipid accumulation in zebrafish liver cells. Comp Biochem Physiol C Toxicol Pharmacol 230:108704

    CAS  Google Scholar 

  • Perumal S, Gopal Samy MV, Subramanian D (2021) Developmental toxicity, antioxidant, and marker enzyme assessment of swertiamarin in zebrafish (Danio rerio). J Biochem Mol Toxicol 35(9):e22843

    CAS  Google Scholar 

  • Piel C, Pouchieu C, Carles C, Béziat B, Boulanger M, Bureau M, Busson A, Grüber A, Lecluse Y, Migault L, Renier M, Rondeau V, Schwall X, Tual S, Agrican G, Pierre L, Baldi I (2019) Agricultural exposures to carbamate herbicides and fungicides and central nervous system tumour incidence in the cohort AGRICAN. Environ Int 130:104876

    CAS  Google Scholar 

  • Santos D, Félix L, Luzio A, Parra S, Cabecinha E, Bellas J, Monteiro SM (2020) Toxicological effects induced on early life stages of zebrafish (Danio rerio) after an acute exposure to microplastics alone or co-exposed with copper. Chemosphere 261:127748

    CAS  Google Scholar 

  • Shen W, Yang G, Guo Q, Lv L, Liu L, Wang X, Lou B, Wang Q, Wang Y (2021) Combined toxicity assessment of myclobutanil and thiamethoxam to zebrafish embryos employing multi-endpoints. Environ Pollut 269:116116

    CAS  Google Scholar 

  • Soffan A, Alghamdi SS, Aldawood AS (2014) Peroxidase and polyphenol oxidase activity in moderate resistant and susceptible Vicia faba induced by Aphis craccivora (Hemiptera: Aphididae) infestation. J Insect Sci 14:285

    CAS  Google Scholar 

  • Song Z, Zhang Y, Zhang H, Rajendran RS, Wang R, Hsiao CD, Li J, Xia Q, Liu K (2020) Isoliquiritigenin triggers developmental toxicity and oxidative stress-mediated apoptosis in zebrafish embryos/larvae via Nrf2-HO1/JNK-ERK/mitochondrion pathway. Chemosphere 246:125727

    CAS  Google Scholar 

  • Sun Y, Cao Y, Tong L, Tao F, Wang X, Wu H, Wang M (2020) Exposure to prothioconazole induces developmental toxicity and cardiovascular effects on zebrafish embryo. Chemosphere 251:126418

    CAS  Google Scholar 

  • Sun D, Chen Q, Zhu B, Zhao H, Duan S (2021) Multigenerational reproduction and developmental toxicity, and HPG axis gene expression study on environmentally-relevant concentrations of nonylphenol in zebrafish. Sci Total Environ 764:144259

    CAS  Google Scholar 

  • Tooby TE, Hursey PA, Alabaster JS (1975) The acute toxicity of 102 pesticides and miscellaneous substances to fish. Chem Ind-London 12:523–526

    Google Scholar 

  • Vu HT, Keough MJ, Long SM, Pettigrove VJ (2017) Toxicological effects of fungicide mixtures on the amphipod Austrochiltonia subtenuis. Environ Toxicol Chem 36(10):2651–2659

    CAS  Google Scholar 

  • Wei H, Wang S, Xu EG, Liu J, Li X, Wang Z (2020) Synergistic toxicity of microcystin-LR and Cu to zebrafish (Danio rerio). Sci Total Environ 713:136393

    CAS  Google Scholar 

  • Wu S, Lei L, Liu M, Song Y, Lu S, Li D, Shi H, Raley-Susman KM, He D (2018) Single and mixture toxicity of strobilurin and SDHI fungicides to Xenopus tropicalis embryos. Ecotoxicol Environ Saf 153:8–15

    CAS  Google Scholar 

  • Xu X, Li CM, WangWangSongWang SJHLHWZJ (2016) Residue characteristics and ecological risk assessment of twenty-nine pesticides in surface water of major river-basin in China. Asian J Ecotoxicol 11(2):347–354 in Chinese

    Google Scholar 

  • Yan S, Wang J, Zhu L, Chen A, Wang J (2015) Toxic effects of nitenpyram on antioxidant enzyme system and DNA in zebrafish (Danio rerio) livers. Ecotoxicol Environ Saf 122:54–60

    CAS  Google Scholar 

  • Yao F, Li Y, Ru H, Wu L, Xiao Z, Ni Z, Chen D, Zhong L (2020) Thyroid disruption and developmental toxicity caused by triphenyltin (TPT) in zebrafish embryos/larvae. Toxicol Appl Pharmacol 394:114957

    CAS  Google Scholar 

  • Zhang C, Wang J, Zhang S, Zhu LS, Du ZK, Wang JH (2017) Acute and subchronic toxicity of pyraclostrobin in zebrafish (Danio rerio). Chemosphere 188:510–516

    CAS  Google Scholar 

  • Zhang C, Zhou T, Xu Y, Du Z, Li B, Wang J, Wang J, Zhu L (2020) Ecotoxicology of strobilurin fungicides. Sci Total Environ 742:140611

    CAS  Google Scholar 

  • Zhao W, Hu N, Ding D, Long D, Li S, Li G, Zhang H (2019) Developmental toxicity and apoptosis in zebrafish embryos induced by low-dose γ-ray irradiation. Environ Sci Pollut Res 26(4):3869–3881

    CAS  Google Scholar 

  • Zhu B, Han J, Lei L, Hua J, Zuo Y, Zhou B (2021) Effects of SiO2 nanoparticles on the uptake of tetrabromobisphenol A and its impact on the thyroid endocrine system in zebrafish larvae. Ecotoxicol Environ Saf 209:111845

    CAS  Google Scholar 

  • Zubrod JP, Bundschuh M, Arts G, Brühl CA, Imfeld G, Knäbel A, Payraudeau S, Rasmussen JJ, Rohr J, Scharmüller A, Smalling K, Stehle S, Schulz R, Schäfer RB (2019) Fungicides: an overlooked pesticide class? Environ Sci Technol 53(7):3347–3365

    CAS  Google Scholar 

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Funding

The research was supported by the Scientific and Technological Innovation Project of Yantai City, China (Grant No. 2020MSGY053) and State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products (Grant No. 2021DG700024-KF202106).

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Authors and Affiliations

  1. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Quality and Standard for Agro-Products, Zhejiang Academy of Agricultural Sciences, HangzhouZhejiang, 310021, China

    Yanhua Wang, Xinju Liu & Jindong Ren

  2. College of Chemistry, Research Institute of Elemento-Organic Chemistry, Nankai University, Tianjin, 300071, China

    Zhongwen Gao

  3. Yantai Academy of Agricultural Sciences, Yantai, 265500, China

    Chuande Liu, Zeqi Lu, Jie Yao & Xuan Liu

  4. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Liangang Mao

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  1. Yanhua Wang
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  2. Zhongwen Gao
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  3. Chuande Liu
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  4. Liangang Mao
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  5. Xinju Liu
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  7. Zeqi Lu
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  8. Jie Yao
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  9. Xuan Liu
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Contributions

Yanhua Wang: conceptualization, methodology, validation, formal analysis, investigation, data curation, writing-original draft, writing—review and editing. Zhongwen Gao: conceptualization, methodology, software, formal analysis, investigation, resources, data curation. Chuande Liu: investigation, data curation. Liangang Mao: investigation, data curation, writing—review and editing. Xinju Liu: writing—review and editing, administration, funding acquisition. Jindong Ren: investigation, resources. Zeqi Lu: conceptualization, methodology. Jie Yao: methodology, software, formal analysis. Xuan Liu: writing—review and editing, supervision, project administration, funding acquisition, corresponding author.

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Correspondence to Xuan Liu.

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Wang, Y., Gao, Z., Liu, C. et al. Mixture toxicity of pyraclostrobine and metiram to the zebrafish (Danio rerio) and its potential mechanism. Environ Sci Pollut Res 30, 44400–44414 (2023). https://doi.org/10.1007/s11356-023-25518-3

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