Abstract
The insecticide fipronil, one of the main pesticides used in Brazil, is often detected in natural aquatic environments, and causes neuronal hyperexcitation by inhibiting GABAergic neurotransmission, leading to putative alterations in behaviour and development. This work sought to analyse the toxicity of formulated Regent® 800WG (80% fipronil) on development (fish embryo toxicity test, FET), morphology, and swimming behaviour of larvae and adults of zebrafish (Danio rerio). FET was performed following OECD236 guidelines at concentrations ranging from 0.002 to 1600 μg.L−1 of formulated Regent® 800WG. Adults were exposed to 0.2, 2 and 20 μg.L−1 of the product for 24 and 96 h, and were submitted to the light-dark, novel tank and swimming endurance tests No lethal parameters were observed in larvae, but in concentrations above 400 µg.L−1, there was shortening of the body axis and decreased swimming behavior. In adults, exposure to the pesticide did not lead to changes in free swimming parameters. However, a marked decrease of swimming endurance was observed at all experimental treatments, although probably not in consequence of energetic depletion, since baseline blood glucose levels and condition factor were similar at all conditions. Furthermore, zebrafish adults did not show their natural preference for the dark environment. The pesticide likely has anxiolytic effects on zebrafish, as well as a compromising effect on locomotor control, illustrating that behavioural changes, which could affect activities on the natural environment, such as escape and predation, may occur even in environmentally relevant concentrations of this pollutant.
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References
Bakshi VP, Kalin NH (2002) Animal models and endophenotypes of anxiety and stress disorder. Neuropsychopharmacol Fifth Gener Prog 62:884–901
Ballesteros ML, Durando PE, Nores ML, Díaz MP, Bistoni MA, Wunderlin DA (2009) Endosulfan induces changes in spontaneous swimming activity and acetylcholinesterase activity of Jenynsia multidentata (Anablepidae, Cyprinodontiformes). Environ Pollut 157:1573–1580. https://doi.org/10.1016/j.envpol.2009.01.001
Beggel S, Werner I, Connon RE, Geist JP (2010) Sublethal toxicity of commercial insecticide formulations and their active ingedients to larval fathead minnow (Pimephales promelas). Sci Total Environ 408(16):3169–3175. https://doi.org/10.1016/j.scitotenv.2010.04.004
Bencan Z, Sledge D, Levin ED (2009) Buspirone, chlordiazepoxide and diazepam effects in a zebra fish model of anxiety. Pharmacol Biochem Behav 94:75–80. https://doi.org/10.1016/j.pbb.2009.07.009
Bevilaqua F, Sachett A, Chitolina R, Garbinato C, Gasparetto H, Marcon M, Mocelin R, Dallegrave E, Conterato G, Piato A, Siebel AM (2020) A mixture of fipronil and fungicides induces alterations on behavioral and oxidative stress parameters in zebrafish. Ecotoxicology 29:140–147. https://doi.org/10.1007/s10646-019-02146-7
Blaser RE, Goldsteinholm K (2012) Depth preference in zebrafish, Danio rerio: Control by surface and substrate cues. Anim Behav 83(4):953–959. https://doi.org/10.1016/j.anbehav.2012.01.014
Brasil (2005) Ministério do Meio Ambiente. Conselho Nacional do Meio Ambiente (CONAMA). Resolução CONAMA No. 357, de 17 de março de 2005.
Brasil (2011) Ministério do Meio Ambiente. Conselho Nacional do Meio Ambiente (CONAMA). Resolução CONAMA No. 430, de 13 de maio de 2011.
Chaulet FC, Barcellos HHA, Fior D, Pompermaier A, Koakoski G, Rosa JGS, Fagundes M, Barcellos LJG (2019) Glyphosate-and fipronil-based agrochemicals and their mixtures change zebrafish behavior. Arch Environ Con Tox 77(3):443–451. https://doi.org/10.1007/s00244-019-00644-7
Clasen B, Loro VL, Cattaneo R, Moraes B, Lópes T, de Avila LA, Zanella R, Reimche GB, Baldisserotto B (2012) Effects of the commercial formulation containing fipronil on the non-target organism Cyprinus carpio: implications for rice-fish cultivation. Ecotoxicol Environ Saf 77:45–51. https://doi.org/10.1016/j.ecoenv.2011.10.001
Coutinho CFB, Tanimoto ST, Galli A, Garbellini GS, Takayama M, Do Amaral RB, Mazo LH, Avaca LA, Machado SAS (2005) Pesticidas: Mecanismo De Ação, Degradação E Toxidez. Pestic Rev Ecotoxicologia e Meio Ambient 15:65–72. https://doi.org/10.5380/pes.v15i0.4469
Dallarés S, Dourado P, Sanahuja I, Solovyev M, Gisbert E, Montemurro N, Torreblanca A, Blázquez M, Solé M (2020) Multibiomarker approach to fipronil exposure in the fish Dicentrarchus labrax under two temperature regimes. Aquat Toxicol 219:105348. https://doi.org/10.1016/j.aquatox.2019.105378
De Esch C, Slieker R, Wolterbeek A, Woutersen R, de Groot D (2012) Zebrafish as potential model for developmental neurotoxicity testing. A mini review. Neurotoxicol Teratol 34:545–553. https://doi.org/10.1016/j.ntt.2012.08.006
Eames SC, Philipson LH, Prince VE, Kinkel MD (2010) Blood sugar measurement in zebrafish reveals dynamics of glucose homeostasis. Zebrafish 7:205–13. https://doi.org/10.1089/zeb.2009.0640
Faimali M, Gambardella C, Costa E, Piazza V, Morgana S, Estévez-Calvar N, Garaventa F (2017) Old model organisms and new behavioral end-points: swimming alteration as an ecotoxicological response. Mar Environ Res128:36–45. https://doi.org/10.1016/j.marenvres.2016.05.006
Farias MS, Schlosser JF, Casali AL, Frantz UG, Rodrigues FA (2010) Qualidade da água utilizada para aplicação de agrotóxicos na região central do Rio Grande do Sul. Ciência Rural 40:1053–1059
Food and Agriculture Organization of the United Nations, (2019). FAOSTAT Database. http://www.fao.org/faostat/en/#data/RP. Accessed 29 June 2019.
Gunasekara AS, Truong T, Goh KS, Spurlock F, Tjeerdema RS (2007) Environmental fate and toxicology of fipronil. Pestic Sci 32(3):189–199. https://doi.org/10.1584/jpestics.R07-02
Gupta SK, Pal AK, Sahu NP, Saharan N, Prakash C, Akhtar MS, Kumar S (2014) Haemato-biochemical responses in Cyprinus carpio (Linnaeus, 1758) fry exposed to sub-lethal concentration of a phenylpyrazole insecticide, fipronil. Proc Natl Acad Sci India B - Biol Sci 84(1):113–122. https://doi.org/10.1007/s40011-013-0201-y
Hellou J (2011) Behavioural ecotoxicology, an “early warning” signal to assess environmental quality. Environ Sci Pollut Res 18:1–11. https://doi.org/10.1007/s11356-010-0367-2
IBAMA, (2019). Annual reports on commercialization of agrotoxics. Retrieved from https://www.ibama.gov.br/agrotoxicos/relatorios-de-comercializacao-de-agrotoxicos#historicodecomercializacao. Accessed 29 June 2019.
Ikeda T, Nagata K, Kono Y, Yeh JZ, Narahashi T (2004) Fipronil modulation of GABAA receptor single-channel currents. Pest Manag Sci 60:487–492. https://doi.org/10.1002/ps.830
Jinguji H, Ohtsu K, Ueda T, Goka K (2018) Effects of short-term, sublethal fipronil and its metabolite on dragonfly feeding activity. PloS ONE 13(7):e0200299. https://doi.org/10.1371/journal.pone.0200299
Jorge LAC, Laumann R, Borges M, Moraes MCB, Cruz RA, Milare BN, Palhares L (2005) Software para Avaliação do Comportamento de Insetos. Circular Técnica 30, 1a Edição - Embrapa CNPDIA, São Carlos, Brazil
Kavaliers M, Choleris E (2001) Antipredator responses and defensive behavior: ecological and ethological approaches for the neurosciences. Neurosci Biobehav Rev 25:577–586. https://doi.org/10.1016/S0149-7634(01)00042-2
Lawrence C (2007) The husbandry of zebrafish (Danio rerio): a review. Aquaculture 269:1–20. https://doi.org/10.1016/j.aquaculture.2007.04.077
Levin ED, Bencan Z, Cerutti DT (2007) Anxiolytic effects of nicotine in zebrafish. Physiol Behav 90:54–58. https://doi.org/10.1016/j.physbeh.2006.08.026
Maximino C, Brito TM, De, Gazolla RA, Tenório R, De RIT (2007) A comparative analysis of the preference for dark environments in five teleosts. Int J Comp Psychol 20(4):351–367. https://doi.org/10.5070/P4204009997
Maximino C, Marques de Brito T, Dias CAGDM, Gouveia A, Morato S (2010a) Scototaxis as anxiety-like behavior in fish. Nat Protoc 5:209–216. https://doi.org/10.1038/nprot.2009.225
Maximino C, de Brito TM, da Silva Batista AW, Herculano AM, Morato S, Gouveia A (2010b) Measuring anxiety in zebrafish: a critical review. Behav Brain Res 214:157–71. https://doi.org/10.1016/j.bbr.2010.05.031
Menezes C, Leitemperger J, Murussi C, Souza M, De, Martha V, Renato BA (2016) Effect of diphenyl diselenide diet supplementation on oxidative stress biomarkers in two species of freshwater fish exposed to the insecticide fipronil. Fish Physiol Biochem 42:1357–1368. https://doi.org/10.1007/s10695-016-0223-5
Norton W, Bally-Cuif L (2010) Adult zebrafish as a model organism for behavioural genetics. BMC Neurosci 11:11–90. https://doi.org/10.1186/1471-2202-11-90
OECD (2013). Test no. 236: Fish Embryo Acute Toxicity (FET), OECD Guidelines for Testing of Chemicals.
Ohlberger J, Staaks G, Van Dijk PLM, Hölker F (2005) Modelling energetic costs of fish swimming. J Exp Zool303(8):657–664. https://doi.org/10.1002/jez.a.181
Parker MO, Millington ME, Combe FJ, Brennan CH (2012) Housing conditions differentially affect physiological and behavioural stress responses of Zebrafish, as well as the responste to anxiolytics. PloS ONE 7:e34992. https://doi.org/10.1371/journal.pone.0034992
Post JR, Lee JA (1996) Metabolic ontogeny of teleost fishes. Can J Fish Aquat Sci 53(4):910–923. https://doi.org/10.1139/f95-278
Qu H, Ma R, Liu D, Gao J, Wang F, Zhou Z, Wang P (2016) Environmental behavior of the chiral insecticide fipronil: enantioselective toxicity, distribution and transformation in aquatic ecosystem. Water Res 105:138–146. https://doi.org/10.1016/j.watres.2016.08.063
Rao JV, Begum G, Pallela R, Usman PK, Rao RN (2005) Changes in behavior and brain acetylcholinesterase activity in Mosquito Fish, Gambusia affinis in response to the sub-lethal exposure to chlorpyrifos. Int J Environ Res Public Health 2(3):478–483
Robinson PD (2009) Behavioural toxicity of organic chemical contaminants in fish: application to ecological risk assessments (ERAs). Can J Fish Aquat Sci 66:1179–1188. https://doi.org/10.1139/f09-069
Saka M, Tada N (2021) Acute and chronic toxicity tests of systemic insecticides, four neonicotinoids and fipronil, using the tadpoles of the western clawed frog Silurana tropicalis. Chemosphere 270:129418. https://doi.org/10.1016/j.chemosphere.2020.129418
Schwarzenbach RP, Escher BI, Hostetter T, Jonhson BC, Von Gunten AU, Wehrli B (2006) The challenge of micropollutants in aquatic systems. Science 313(5790):1072–1077. https://doi.org/10.1126/science.1127291
Scott GR, Sloman KA (2004) The effects of environmental pollutants on complex fish behaviour: integrating behavioural and physiological indicators of toxicity. Aquat Toxicol 68:369–392. https://doi.org/10.1016/j.aquatox.2004.03.016
Serra EL, Medalha CC, Mattioli R (1999) Natural preference of zebrafish (Danio rerio) for a dark environment. Brazilian J Med Biol Res 32:1551–1553. https://doi.org/10.1590/S0100-879X1999001200016
Silva DRO, Avila LA, Agostinetto D, Magro TD, Oliveira E, Zanella R, Noldin JA (2009) Monitoramento de agrotóxicos em águas superficiais de regiões orizícolas no sul do Brasil. Ciência Rural 39:2383–2389. https://doi.org/10.1590/S0103-84782009000900001
Silva LC, Moreira RA, Pinto TJ, Ogura AP, Yoshii MP, Lopes LF, Montagner CC, Goulart BV, Daam MA, Espíndola EL (2020) Acute and chronic toxicity of 2,4-D and fipronil formulations (individually and in mixture) to the Neotropical cladoceran Ceriodaphnia silvestrii. Ecotoxicology 29(9):1462–1475
Simonato JD, Mela M, Doria HB, Guiloski IC, Randi MAF, Carvalho PSM, Meletti PC, Silva HC, Assis D, Bianchini A, Martinez CBR (2016) Biomarkers of waterborne copper exposure in the Neotropical fish Prochilodus lineatus. Aquat Toxicol 170:31–41. https://doi.org/10.1016/j.aquatox.2015.11.012
Simon-Delso N, Amaral-Rogers V, Belzunces LP, Bonmatin JM, Chagnon M, Downs C, Furlan L, Gibbons DW, Giorio C, Girolami V, Goulson D, Kreutzweiser DP, Krupke CH, Liess M, Long E, Mcfield M, Mineau P, Mitchell EA, Morrissey CA, Noome DA, Pisa L, Settele J, Stark JD, Tapparo A, Van Dyck H, Van Praagh J, Van Der Sluijs JP, Whitehorn PR, Wiemers M (2015) Systemic insecticides (Neonicotinoids and fipronil): trends, uses, mode of action and metabolites. Environ Sci Pollut Res 22:5–34. https://doi.org/10.1007/s11356-014-3470-y
Steenbergen PJ, Richardson MK, Champagne DL (2011) Patterns of avoidance behaviours in the light/dark preference test in young juvenile zebrafish: a pharmacological study. Behav Brain Res 222:15–25. https://doi.org/10.1016/j.bbr.2011.03.025
Srean P, Almeida D, Rubio-Gracia F, Luo Y, García-Berthou E (2017) Effects of size and sex on swimming performance and metabolism of invasive Mosquitofish Gambusia holbrooki. Ecol Freshw Fish 26(3):424–433
Stehr CM, Linbo TL, Incardona JP, Scholz NL (2006) The developmental neurotoxicity of fipronil: Notochord degeneration and locomotor defects in zebrafish embryos and larvae. Toxicol Sci 92:270–278. https://doi.org/10.1093/toxsci/kfj185
Stemple DL (2005) Structure and function of the notochord: an essential organ for chordate development. Development 132:2503–2512. https://doi.org/10.1242/dev.01812
Vieira CED, Costa PG, Caldas SS, Tesser ME, Risso WE, Escarrone ALV, Primel EG, Bianchini A, Maritinez CBR (2019) An integrated approach in subtropical agro-ecosystems: active biomonitoring, environmental contaminants, bioaccumulation, and multiple biomarkers in fish. Sci Tot Environ 666:508–524. https://doi.org/10.1016/j.scitotenv.2019.02.209
Vieira LR, Gravato C, Soares AMVM, Morgado F, Guilhermino L (2009) Acute effects of copper and mercury on the estuarine fish Pomatoschistus microps: Linking biomarkers to behaviour. Chemosphere 76:1416–1427. https://doi.org/10.1016/j.chemosphere.2009.06.005
Vignet C, Bégout M-L, Péan S, Lyphout L, Leguay D, Cousin X (2013) Systematic screening of behavioral responses in two Zebrafish Strains. Zebrafish 10:365–375. https://doi.org/10.1089/zeb.2013.0871
Wagner SD, Kurobe T, Hammock BG, Lam CH, Wu G, Vasylieva N, Gee SJ, Hammock BD, Teh SJ (2017) Developmental effects of fipronil on Japanese Medaka (Oryzias latipes). Chemosphere 166:511–520. https://doi.org/10.1016/j.chemosphere.2016.09.069
Wang X, Zhou S, Ding X, Zhu G, Guo J (2010) Effect of triazophos, fipronil and their mixture on miRNA expression in adult zebrafish. J Environ Sci Health B 45:648–57. https://doi.org/10.1080/03601234.2010.502435
Wu H, Gao C, Guo Y, Zhang Y, Zhang J, Ma E (2014) Acute toxicity and sub-lethal effects of fipronil on detoxification enzymes in juvenile zebrafish (Danio rerio). Pestic Biochem Physiol 115:9–14. https://doi.org/10.1016/j.pestbp.2014.07.010
Yoshida M, Nagamine M, Uematsu K (2005) Comparison of behavioral responses to a novel environment between three teleosts, bluegill Lepomis macrochirus, crucian carp Carassius langsdorfii, and goldfish Carassius auratus. Fish Sci 71:314–319. https://doi.org/10.1111/j.1444-2906.2005.00966.x
Zaluski R, Kadri SM, Alonso DP, Ribolla PEM, Orsi RO (2015) Fipronil promotes motor and behavioral changes in honey bees (Apis mellifera) and affects the development of colonies exposed to sublethal doses. Environ Toxicol Chem 34(5):1062–1069. https://doi.org/10.1002/etc.2889
Zhu F, Wigh A, Friedrich T, Devaux A, Bony S, Nugegoda D, Kaslin J, Wlodkowic D (2015) Automated Lab-on-a-chip technology for fish embryo toxicity tests performed under continuous microperfusion (µFET). Environ Sci Technol 49:14570–14578. https://doi.org/10.1021/acs.est.5b03838
Acknowledgements
The authors thank the State University of Londrina and the Laboratory of Animal Ecophysiology for the technical support. ALRC would like to thank Ana Flávia G Campos and Gustavo W Vituri for their assistance recording behavioural tests; and Vinicius Popolin and Caísa Batista for their support with the FET analyses.
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PCM developed the original idea and experimental design, supervised the experiments and data analyses, and edited the paper. ALRC performed all experiments, all analyses, and calculations, and wrote the paper. JDS reviewed the paper, and contributed to the discussion and data interpretation.
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The protocols and experiments here described were approved by the Bioethics Committee of the State University of Londrina, license no. 12766.2018.54.
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Cuenca, A.L.R., Simonato, J.D. & Meletti, P.C. Acute exposure of embryo, larvae and adults of Danio rerio to fipronil commercial formulation reveals effects on development and motor control. Ecotoxicology 31, 114–123 (2022). https://doi.org/10.1007/s10646-021-02497-0
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DOI: https://doi.org/10.1007/s10646-021-02497-0