Abstract
Isopyrazam (IPZ) is a broad spectrum succinate dehydrogenase inhibitor fungicide. Little is known about its potential ecological risks of aquatic organisms recently. The present study examined the embryonic development effects of zebrafish exposed to IPZ under static condition using a fish embryo toxicity test. The lowest observed effect concentration of IPZ was 0.025 mg/L in 4-day exposure. Developmental abnormalities, including edema, small head deformity, body deformation and decreased pigmentation, and mortality were observed in zebrafish embryos of 0.05 mg/L and higher concentrations, which shown concentration dependency. The heart rate of zebrafish was disrupted by IPZ. Moreover, enzyme and gene experiments shown that IPZ exposure caused oxidative stress of zebrafish. Furthermore, it induced a decrease of succinate dehydrogenase (SDH) enzyme activity and gene transcription level in zebrafish larvae. It can be speculated that IPZ may have a lethal effect on zebrafish, which is accompanied by decreased SDH activity, oxidative stress, and abnormality. These results provide toxicological data about the IPZ on aquatic non-target organisms, which could be useful for further understanding potential environmental risks.
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Abad-Fuentes A, Ceballos-Alcantarilla E, Mercader JV, Agulló C, Abad-Somovilla A, Esteve-Turrillas FA (2015) Determination of succinate-dehydrogenase-inhibitor fungicide residues in fruits and vegetables by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 407(14):4207–4211
Andrade TS, Henriques JF, Almeida AR, Machado AL, Koba O, Giang PT, Soares AMVM, Domingues I (2016) Carbendazim exposure induces developmental, biochemical and behavioural disturbance in zebrafish embryos. Aquat Toxicol 170:390–399
APVMA (2018) Australian pesticides and veterinary medicines authority. Proposed registration of Seguris Flexi Fungicide containing the new active constituent isopyrazam. https://apvma.gov.au/node/28996. Accessed 21 April 2018
Avenot HF, Michailides TJ (2010) Progress in understanding molecular mechanisms and evolution of resistance to succinate dehydrogenase inhibiting (SDHI) fungicides in phytopathogenic fungi. Crop Prot 29(7):643–651
Avetta P, Marchetti G, Minella M, Pazzi M, De Laurentiis E, Maurino V, Minero C, Vione D (2014) Phototransformation pathways of the fungicide dimethomorph ((E, Z) 4-[3-(4-chlorophenyl)-3-(3, 4-dimethoxyphenyl)-1-oxo-2-propenyl] morpholine), relevant to sunlit surface waters. Sci Total Environ 500:351–360
Blahová J, Plhalová L, Hostovský M, Divišová L, Dobšíková R, Mikulíková I, Stěpánová S, Svobodová Z (2013) Oxidative stress responses in zebrafish danio rerio after subchronic exposure to atrazine. Food Chem Toxicol 61(6):82–85
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
Braunbeck T, Böttcher M, Hollert H, Kosmehl T, Lammer E, Leist E, Rudolf M, Seitz N (2005) Towards an alternative for the acute fish LC50 test in chemical assessment: the fish embryo toxicity test goes multi-species—an update. ALTEX 22(2):87–102
Braunbeck T, Kais B, Lammer E, Otte J, Schneider K, Stengel D, Strecker R (2015) The fish embryo test (FET): origin, applications, and future. Environ Sci Pollut Res 22(21):16247–16261
Bunzel K, Schäfer RB, Thrän D, Kattwinkel M (2015) Pesticide runoff from energy crops: a threat to aquatic invertebrates? Sci Total Environ 537:187–196
Busquet F, Strecker R, Rawlings JM, Belanger SE, Braunbeck T, Carr GJ, Cenijn P, Fochtman P, Gourmelon A, Hübler N, Kleensang A, Knöbel M, Kussatz C, Legler J, Lillicrap A, Martínez-Jerónimo F, Polleichtner C, Rzodeczko H, Salinas E, Schneider KE, Scholz S, van den Brandhof EJ, van der Ven LT, Walter-Rohde S, Weigt S, Witters H, Halder M (2014) OECD validation study to assess intra-and inter-laboratory reproducibility of the zebrafish embryo toxicity test for acute aquatic toxicity testing. Regul Toxicol Pharmacol 69(3):496–511
Buttke TM, Sandstrom PA (1994) Oxidative stress as a mediator of apoptosis. Immunol Today 15(1):7–10
Cambier S, Benard G, Mesmer-Dudons N, Gonzalez P, Rossignol R, Brethes D, Bourdineaud JP (2009) At environmental doses, dietary methylmercury inhibits mitochondrial energy metabolism in skeletal muscles of the zebra fish (Danio rerio). Int J Biochem Cell Biol 41(4):791–799
Chen JN, Haffter P, Odenthal J, Vogelsang E, Brand M, Van Eeden FJ, Furutani-Seiki M, Granato M, Hammerschmidt M, Heisenberg CP, Jiang YJ, Kane DA, Kelsh RN, Mullins MC, Nüsslein-Volhard C (1996) Mutations affecting the cardiovascular system and other internal organs in zebrafish. Development 123(1):293–302
De Luca E, Zaccaria GM, Hadhoud M, Rizzo G, Ponzini R, Morbiducci U, Santoro MM (2014) ZebraBeat: a flexible platform for the analysis of the cardiac rate in zebrafish embryos. Sci Rep 4:649–652
Di Giulio RT, Washburn PC, Wenning RJ, Winston GW, Jewell CS (1989) Biochemical responses in aquatic animals: a review of determinants of oxidative stress. Environ Toxicol Chem 8(12):1103–1123
Driessen M, Vitins AP, Pennings JL, Kienhuis AS, van de Water B, van der Ven LT (2015) A transcriptomics-based hepatotoxicity comparison between the zebrafish embryo and established human and rodent in vitro and in vivo models using cyclosporine A, amiodarone and acetaminophen. Toxicol Lett 232(2):403–412
EPA (2004) United States Environmental Protection Agency. Chemical hazard classification and labeling: comparison of OPP requirements and the GHS. https://www.epa.gov/sites/production/files/2015-09/documents/ghscriteria-summary.pdf. Accessed 11 Dec 2017
EPA (2017) United States Environmental Protection Agency. Isopyrazam Fact Sheet. https://www3.epa.gov/pesticides/chem_search/reg_actions/pending/fs_PC-129222_05-Oct-11.pdf. Accessed 20 April 2018
Fernández D, Voss K, Bundschuh M, Zubrod JP, Schäfer RB (2015) Effects of fungicides on decomposer communities and litter decomposition in vineyard streams. Sci Total Environ 533:40–48
Fong HCH, Ho JCH, Cheung AHY, Lai KP, Tse WKF (2016) Developmental toxicity of the common UV filter, benophenone-2, in zebrafish embryos. Chemosphere 164:413–420
FRAC (2017) FRAC Code List 2017: Fungicides sorted by mode of action (including FRAC Code numbering). http://www.frac.info/docs/default-source/publications/frac-code-list/frac-code-list-2017-final.pdf. Accessed 11 December 2017
Glickman NS, Yelon D (2002) Cardiac development in zebrafish: coordination of form and function. Semin Cell Dev Biol 13(6):507–513
Gulkowska A, Buerge IJ, Poiger T (2014) Online solid phase extraction LC–MS/MS method for the analysis of succinate dehydrogenase inhibitor fungicides and its applicability to surface water samples. Anal Bioanal Chem 406(25):6419–6427
Han Y, Liu T, Wang J, Wang J, Zhang C, Zhu L (2016) Genotoxicity and oxidative stress induced by the fungicide azoxystrobin in zebrafish (Danio rerio) livers. Pestic Biochem Physiol 133:13–19
Hand LH, Moreland HJ (2014) Surface water mineralization of isopyrazam according to OECD 309: observations on implementation of the new data requirement within agrochemical regulation. Environ Toxicol Chem 33(3):516–524
Hand LH, Oliver RG (2010) The behavior of isopyrazam in aquatic ecosystems: implementation of a tiered investigation. Environ Toxicol Chem 29(12):2702–2712
Hatefi Y (1985) The mitochondrial electron transport and oxidative phosphorylation system. Annu Rev Biochem 54(1):1015–1069
He L, Cui K, Ma D, Shen R, Huang X, Jiang J, Mu W, Liu F (2017) Activity, translocation and persistence of isopyrazam for controlling cucumber powdery mildew. Plant Dis 101(7):1139–1144
Hederstedt L, Rutberg L (1981) Succinate dehydrogenase—a comparative review. Microbiol Rev 45(4):542–555
Hermann AC, Millard PJ, Blake SL, Kim CH (2004) Development of a respiratory burst assay using zebrafish kidneys and embryos. J Immunol Methods 292(1):119–129
Hermsen SA, van den Brandhof EJ, van der Ven LT, Piersma AH (2011) Relative embryotoxicity of two classes of chemicals in a modified zebrafish embryotoxicity test and comparison with their in vivo potencies. Toxicol in Vitro 25(3):745–753
Hill AJ, Teraoka H, Heideman W, Peterson RE (2005) Zebrafish as a model vertebrate for investigating chemical toxicity. Toxicol Sci 86(1):6–19
Jin Y, Zhang X, Shu L, Chen L, Sun L, Qian H, Liu W, Fu Z (2010) Oxidative stress response and gene expression with atrazine exposure in adult female zebrafish (Danio rerio). Chemosphere 78(7):846–852
Jones DP (2006) Redefining oxidative stress. Antioxid Redox Signal 8(9–10):1865–1879
Keon JP, White GA, Hargreaves JA (1991) Isolation, characterization and sequence of a gene conferring resistance to the systemic fungicide carboxin from the maize smut pathogen, Ustilago maydis. Curr Genet 19(6):475–481
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408
Lushchak VI (2011) Environmentally induced oxidative stress in aquatic animals. Aquat Toxicol 101(1):13–30
Massarsky A, Kozal JS, Di Giulio RT (2017) Glutathione and zebrafish: old assays to address a current issue. Chemosphere 168:707–715
McCord JM, Fridovich I (1969) Superoxide dismutase an enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244(22):6049–6055
Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7(9):405–410
Mnif W, Hassine AIH, Bouaziz A, Bartegi A, Thomas O, Roig B (2011) Effect of endocrine disruptor pesticides: a review. Int J Environ Res Public Health 8(6):2265–2303
Moreno-González R, Campillo JA, García V, León VM (2013) Seasonal input of regulated and emerging organic pollutants through surface watercourses to a Mediterranean coastal lagoon. Chemosphere 92(3):247–257
Moura MA, Oliveira R, Jonsson CM, Domingues I, Soares AM, Nogueira AJ (2017) The sugarcane herbicide ametryn induces oxidative stress and developmental abnormalities in zebrafish embryos. Environ Sci Pollut Res:1–10
Mu X, Chai T, Wang K, Zhu L, Huang Y, Shen G, Li Y, Li X, Wang C (2016) The developmental effect of difenoconazole on zebrafish embryos: a mechanism research. Environ Pollut 212:18–26
Niemann S, Müller U (2000) Mutations in SDHC cause autosomal dominant paraganglioma, type 3. Nat Genet 26(3):268–270
Noyes PD, Garcia GR, Tanguay RL (2016) Zebrafish as an in vivo model for sustainable chemical design. Green Chem 18(24):6410–6430
OECD (2013) Fish embryos acute toxicity (FET) test, Test Guideline No. 236, OECD guidelines for testing of chemicals. Organization for Economic Co-operation and Development, Paris
Olivari FA, Hernández PP, Allende ML (2008) Acute copper exposure induces oxidative stress and cell death in lateral line hair cells of zebrafish larvae. Brain Res 1244:1–12
Oyedotun KS, Lemire BD (2004) The quaternary structure of the saccharomyces cerevisiae succinate dehydrogenase homology modeling, cofactor docking, and molecular dynamics simulation studies. J Biol Chem 279(10):9424–9431
Plummer SM, Wright J, Currie RA (2018) Dose-dependent effects on rat liver miRNAs 200a/b and 429: potential early biomarkers of liver carcinogenesis. Toxicol Rep 5:309–313
Rasmussen JJ, McKnight US, Loinaz MC, Thomsen NI, Olsson ME, Bjerg PL, Binning PJ, Kronvang B (2013) A catchment scale evaluation of multiple stressor effects in headwater streams. Sci Total Environ 442:420–431
Reilly TJ, Smalling KL, Orlando JL, Kuivila KM (2012) Occurrence of boscalid and other selected fungicides in surface water and groundwater in three targeted use areas in the United States. Chemosphere 89(3):228–234
Rouquie D, Tinwell H, Blanck O, Schorsch F, Geter D, Wason S, Bars R (2014) Thyroid tumor formation in the male mouse induced by fluopyram is mediated by activation of hepatic CAR/PXR nuclear receptors. Regul Toxicol Pharmacol 70(3):673–680
Sarmah S, Marrs JA (2016) Zebrafish as a vertebrate model system to evaluate effects of environmental toxicants on cardiac development and function. Int J Mol Sci 17:2123
Scalliet G, Bowler J, Luksch T, Kirchhofer-Allan L, Steinhauer D, Ward K, Niklaus M, Verras A, Csukai M, Daina A, Fonné-Pfister R (2012) Mutagenesis and functional studies with succinate dehydrogenase inhibitors in the wheat pathogen Mycosphaerella graminicola. PLoS One 7(4):e35429
Selderslaghs IWT, Blust R, Witters HE (2012) Feasibility study of the zebrafish assay as an alternative method to screen for developmental toxicity and embryotoxicity using a training set of 27 compounds. Reprod Toxicol 33(2):142–154
Sies H (1986) Biochemistry of oxidative stress. Angew Chem Int Ed 25(12):1058–1071
Sies H (1999) Glutathione and its role in cellular functions. Free Radic Biol Med 27(9):916–921
Sies H (2015) Oxidative stress: a concept in redox biology and medicine. Redox Biol 4:180–183
Smalling KL, Kuivila KM, Orlando JL, Phillips BM, Anderson BS, Siegler K, Huntb JW, Hamilton M (2013) Environmental fate of fungicides and other current-use pesticides in a central California estuary. Mar Pollut Bull 73(1):144–153
Song Y, Zhang Z, Chen L, He L, Lu H, Ren Y, Mu W, Feng L (2016) Baseline sensitivity of botrytis cinerea to the succinate dehydrogenase inhibitor isopyrazam and efficacy of this fungicide. Plant Dis 100(7):1314–1320
Stengel D, Wahby S, Braunbeck T (2018) In search of a comprehensible set of endpoints for the routine monitoring of neurotoxicity in vertebrates: sensory perception and nerve transmission in zebrafish (Danio rerio) embryos. Environ Sci Pollut Res:1–19
Stowe DF, Camara AKS (2009) Mitochondrial reactive oxygen species production in excitable cells: modulators of mitochondrial and cell function. Antioxid Redox Signal 11(6):1373–1414
Strähle U, Scholz S, Geisler R, Greiner P, Hollert H, Rastegar S, Schumacher A, Selderslaghs I, Weiss C, Witters H, Braunbeck T (2012) Zebrafish embryos as an alternative to animal experiments—a commentary on the definition of the onset of protected life stages in animal welfare regulations. Reprod Toxicol 33(2):128–132
Tanabe A, Mitobe H, Kawata K, Yasuhara A, Shibamoto T (2001) Seasonal and spatial studies on pesticide residues in surface waters of the Shinano River in Japan. J Agric Food Chem 49(8):3847–3852
Tinwell H, Rouquié D, Schorsch F, Geter D, Wason S, Bars R (2014) Liver tumor formation in female rat induced by fluopyram is mediated by CAR/PXR nuclear receptor activation. Regul Toxicol Pharmacol 70(3):648–658
Townsend DM, Tew KD (2003) The role of glutathione-s-transferase in anti-cancer drug resistance. Oncogene 22(47):7369–7375
Tsuda T, Nakamura T, Inoue A, Tanaka K (2009) Pesticides in water and sediment from littoral area of Lake Biwa. Bull Environ Contam Toxicol 82(6):683–689
Tu W, Niu L, Liu W, Xu C (2013) Embryonic exposure to butachlor in zebrafish (Danio rerio): endocrine disruption, developmental toxicity and immunotoxicity. Ecotoxicol Environ Saf 89:189–195
Uttara B, Singh AV, Zamboni P, Mahajan RT (2009) Oxidative stress and neurodegenerative diseases: a review of upstream and downstream antioxidant therapeutic options. Curr Neuropharmacol 7(1):65–74
Wang Y, Yang G, Dai D, Xu Z, Cai L, Wang Q, Yu Y (2017) Individual and mixture effects of five agricultural pesticides on zebrafish (Danio rerio) larvae. Environ Sci Pollut Res 24(5):4528–4536
Westerfield M (1995) The zebrafish book: a guide for the laboratory use of zebrafish (Brachydanio rerio). University of Oregon Press, Eugene
Wiegand C, Krause E, Steinberg C, Pflugmacher S (2001) Toxicokinetics of atrazine in embryos of the zebrafish (Danio rerio). Ecotoxicol Environ Saf 49(3):199–205
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
Xiao W, Sarsour EH, Wagner BA, Doskey CM, Buettner GR, Domann FE, Goswami PC (2016) Succinate dehydrogenase activity regulates PCB3-quinone-induced metabolic oxidative stress and toxicity in HaCaT human keratinocytes. Arch Toxicol 90(2):319–332
Yang Y, Liu W, Mu X, Qi S, Fu B, Wang C (2016a) Biological response of zebrafish embryos after short-term exposure to thifluzamide. Sci Rep 6:38485
Yang Y, Qi S, Wang D, Wang K, Zhu L, Chai T, Wang C (2016b) Toxic effects of thifluzamide on zebrafish (Danio rerio). J Hazard Mater 307:127–136
Yankovskaya V, Horsefield R, Törnroth S, Luna-Chavez C, Miyoshi H, Léger C, Byrne B, Cecchini G, Iwata S (2003) Architecture of succinate dehydrogenase and reactive oxygen species generation. Science 299(5607):700–704
Yoshida M, Inoue K, Takahashi M (2015) Predictive modes of action of pesticides in uterine adenocarcinoma development in rats. J Toxicol Pathol 28(4):207–216
Zhu B, Liu L, Gong YX, Ling F, Wang GX (2014) Triazole-induced toxicity in developing rare minnow (Gobiocypris rarus) embryos. Environ Sci Pollut R 21(23):13625–13635
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The authors sincerely thank Jason Martin (United States Department of Agriculture) for paper improvement.
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This study received grants from the Project of Science and Technology by Zhejiang Food and Drug Administration (grant number SP201717), the Project by Science Technology Department of Zhejiang Province (grant number LGF18B070003), and the National Natural Science Foundation of China (grant number 21277126).
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Yao, H., Xu, X., Zhou, Y. et al. Impacts of isopyrazam exposure on the development of early-life zebrafish (Danio rerio). Environ Sci Pollut Res 25, 23799–23808 (2018). https://doi.org/10.1007/s11356-018-2449-5
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DOI: https://doi.org/10.1007/s11356-018-2449-5