Abstract
In Brazilian fish farms, trichlorfon has been widely used to control acanthocephalan infections in Colossoma macropomum. Toxicity tests were conducted to estimate the median lethal concentration (LC50–96 h) and evaluate the effects of trichlorfon on acetylcholinesterase (AChE) and glutathione S-transferase (GST) from different tissues of C. macropomum. The LC50–96 h of trichlorfon was estimated to be 0.87 mg L−1. In the sublethal toxicity tests, concentrations of 0.26 mg L−1 (30% of LC50–96 h) and 0.43 mg L−1 (50% of LC50–96 h) were used. AChE and GST activities were measured in the brain, muscle, intestine, and liver. In vitro studies were conducted to estimate the kinetic properties and half-maximal inhibitory concentration (IC50) values of AChE in the brain and muscle for trichlorfon. In the sublethal toxicity experiments, inhibition of more than 90% of AChE in the brain, muscle, and intestine was observed. However, the activity of GST did not vary in any of the tissues studied. This finding suggests that trichlorfon is not metabolised by this enzyme. The in vitro assay results suggest that trichlorfon tends to be a classic uncompetitive inhibitor of AChE in both the brain and muscle, since Km and Vmax values decrease, while the slope remains unchanged. The IC50 values of muscle AChE are lower than those of the brain. All these results show that C. macropomum has low tolerance to this pesticide and suggest that brain AChE can be used as a biochemical biomarker, while muscle AChE may be used as an indicator of mortality in toxicological studies.
Similar content being viewed by others
References
Aguiar JP, Fazzi-Gomes PF, Hamoy IG, Santos SEB, Sampaio I (2018) Tracing individuals and populations of the tambaqui, Colossoma macropomum (Cuvier, 1818), from Brazilian hatcheries using microsatellite markers. J Sci Food Agric 99:2998–3004. https://doi.org/10.1002/jsfa.9513
Albendín G, Arellano JM, Manuel-Vez MP, Sarasquete C, Arufe MI (2017) Characterization and in vitro sensitivity of cholinesterases of gilthead seabream (Sparus aurata) to organophosphate pesticides. Fish Physiol Biochem 43:455–464. https://doi.org/10.1007/s10695-016-0299-y2016
Allocati N, Masulli M, Di Ilio C, Federici L (2018) Glutathione transferases: substrates, inhibitors and pro-drugs in cancer and neurodegenerative diseases. Oncogenesis 7:1–15. https://doi.org/10.1038/s41389-017-0025-3
Araújo MC, Assis CRD, Silva LC, Machado DC, Silva KCC, Lima AVA, Carvalho-Jr LB, Bezerra RS, Oliveira MBM (2016) Brain acetylcholinesterase of jaguar cichlid (Parachromis managuensis): from physicochemical and kinetic properties to its potential as biomarker of pesticides and metal ions. Aquat Toxicol 177:182–189. https://doi.org/10.1016/j.aquatox.2016.05.019
Aride PHR, Roubach R, Val AL (2007) Tolerance response of tambaqui Colossoma macropomum (Cuvier) to water pH. Aquac Res 38:588–594. https://doi.org/10.1111/j.1365-2109.2007.01693.x
Assis CRD, Castro PF, Amaral IPG, Carvalho MEVM, Carvalho-Jr LB, Bezerra RS (2010) Characterization of acetylcholinesterase from the brain of the Amazonian tambaqui (Colossoma macropomum) and in vitro effect of organophosphorus and carbamate pesticides. Environ Toxicol Chem 29:2243–2248. https://doi.org/10.1002/etc.272
Assis CRD, Linhares AG, Oliveira VM, França RCP, Maciel-Carvalho EVM, Bezerra RS, Carvalho-Jr LB (2012) Comparative effect of pesticides on brain acetylcholinesterase in tropical fish. Sci. Total Environ 441:141–150. https://doi.org/10.1016/j.scitotenv.2012.09.058
Baldissera MD, Souza CF, Descovi SN, Zanella R, Prestes OD, Matos AFIM, Silva AS, Baldisserotto B, Gris A, Mendes RE (2019) Disturbance of energetic homeostasis and oxidative damage provoked by trichlorfon as relevant toxicological mechanisms using silver catfish as experimental model. Chem Biol Interact 299:94–100. https://doi.org/10.1016/j.cbi.2018.11.015
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. https://doi.org/10.1006/abio.1976.9999
Bretaud S, Toutant JP, Saglio P (2000) Effects of carbofuran, diuron, and nicosulfuron on acetylcholinesterase activity in goldfish (Carassius auratus). Ecotoxicol Environ Saf 47:117–124. https://doi.org/10.1006/eesa.2000.1954
Chandrasekara HU, Pathiratne A (2005) Acetylcholinesterase inhibition and haematological alterations induced in common carp, (Cyprinus carpio) following exposure to low concentrations of Trichlorfon. J Aquacult Res 36:146–150. https://doi.org/10.1111/j.1365-2109.2004.01197.x
Chang CC, Lee PP, Liu CH, Cheng W (2006) Trichlorfon, an organophosphorus insecticide, depresses the immune responses and resistance to Lactococcus garvieae of the giant freshwater prawn Macrobrachium rosenbergii. Fish Shellfish Immunol 20:574–585. https://doi.org/10.1016/j.fsi.2005.06.012
Chapadense PFG, Castro FJ, Almeida JA, Moron SE (2009) Toxicity of atrazine herbicide in Colossoma macropomum. Rev Bras Saude Prod An 10:398–405
Coelho S, Oliveira R, Pereira S, Musso C, Domingues I, Bhujel RC, Soares AM, Nogueira AJ (2011) Assessing lethal and sub-lethal effects of trichlorfon on different trophic levels. Aquat Toxicol 103:191–198. https://doi.org/10.1016/j.aquatox.2011.03.003
Colovic MB, Krstic DZ, Lazarevic-Pasti TD, Bondzic AM, Vasic VM (2013) Acetylcholinesterase inhibitors: pharmacology and toxicology. Curr Neuropharmacol 11:315–335. https://doi.org/10.2174/1570159X11311030006
Cunha FS, Sousa NC, Santos RFB, Meneses JO, Couto MVS, Almeida FTC, Filho JGS, Carneiro PCF, Maria AN, Fujimoto RY (2018) Deltamethrin-induced nuclear erythrocyte alteration and damage to the gills and liver of Colossoma macropomum. Environ Sci Pollut Res 25:15102–15110. https://doi.org/10.1007/s11356-018-1622-1
Ellman GL, Courtney D, Andres V Jr, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95. https://doi.org/10.1016/0006-2952(61)90145-9
FAO (2007) Pesticide residues in food 2007. Joint FAO/WHO meeting on pesticide residues. Food and Agriculture Organization, Rome, Italy. http://www.fao.org/3/a-a1556e.pdf. Accessed 13 February 2019
Fernandes LS, Emerick GL, Santos NAG, Paula ES, Barbosa F Jr, Santos AC (2015) In vitro study of the neuropathic potential of the organophosphorus compounds trichlorfon and acephate. Toxicol in Vitro 29:522–528. https://doi.org/10.1016/j.tiv.2015.01.001
Ferrari A, Venturino A, D’Angelo AMP (2007) Muscular and brain cholinesterase sensitivities to azinphos methyl and carbaryl in the juvenile rainbow trout Oncorhynchus mykiss. Comp Biochem Physiol C Toxicol Pharmacol 146:308–313. https://doi.org/10.1016/j.cbpc.2007.04.002
Ferreira MKA, Silva AW, Silva FCO, Holanda CLA, Barroso SM, Lima JDR, Vieira-Neto AE, Campos AR, Bandeira PN, Santos HS, Lemos TLG, Siqueira SMC, Magalhães FEA, Menezes JESA (2019) Anxiolytic-like effect of chalcone N-{4′-[(E)-3-(4-fluorophenyl)-1-(phenyl) prop-2-en-1-one]} acetamide on adult zebrafish (Danio rerio): involvement of the GABAergic system. Behav Brain Res. https://doi.org/10.1016/j.bbr.2019.03.040
Glisic B, Mihaljevic I, Popovic M, Zaja R, Loncar J, Fent K, Kovacevic R, Smital T (2015) Characterization of glutathione-S-transferases in zebrafish (Danio rerio). Aquat Toxicol 158:50–62. https://doi.org/10.1016/j.aquatox.2014.10.013
Gomes ALS, Coelho-Filho JG, Silva WV, Oliveira MIB, Bernardino G, Costa JI (2017) The impact of Neoechinorhynchus Buttnerae (Golvan, 1956) (Eoacanthocephala: Neoechinorhynchidae) outbreaks on productive and economic performance of the tambaqui Colossoma macropomum (Cuvier, 1818) reared in pounds. Lat Am J Aquat Res 45:496–500. https://doi.org/10.3856/vol45-issue2-fulltext-25
Guimarães ATB, Calil P (2008) Growth evaluation of Oreochromis niloticus (Cichlidae, Neopterygii) exposed to trichlorfon. Braz Arch Biol Technol 51:323–332. https://doi.org/10.1590/S1516-89132008000200012
Lima MG, Silva RX, Silva SN, Rodrigue LSS, Oliveira KR, Batista EJO, Maximino C, Herculano AM (2016) Time-dependent sensitization of stress responses in zebrafish: a putative model for post-traumatic stress disorder. Behav Process 128:70–82. https://doi.org/10.1016/j.beproc.2016.04.009
Lourenço FS, Morey GAM, Malta JCO (2018) The development of Neoechinorhynchus buttnerae (Eoacanthocephala: Neoechinorhynchidae) in its intermediate host Cypridopsis vidua in Brazil. Acta Parasitol 63:354–359. https://doi.org/10.1515/ap-2018-0040
Mataqueiro MI, Nakaghi LSO, Souza JP, Cruz C, Oliveira GH, Urbinati EC (2009) Histopathological changes in the gill, liver and kidney of pacu (Piaractus mesopotamicus, Holmberg, 1887) exposed to various concentrations of trichlorfon. J Appl Ichthyol 25:124–127. https://doi.org/10.1111/j.1439-0426.2008.01160.x
OECD (2010) Short guidance on the threshold approach for acute fish toxicity. Series on testing and assessment No. 126, OECD, Paris. https://www.oecd.org/chemicalsafety/testing/40985084.pdf. Accessed 22 August 2018
Osten JR, Ortíz-Arana A, Guilhermino L, Soares AM (2005) In vivo evaluation of three biomarkers in the mosquitofish (Gambusia yucatana) exposed to pesticides. Chemosphere 58:627–636. https://doi.org/10.1016/j.chemosphere.2004.08.065
Peixe Br (2019) Brazilian Fish Report FISH BR 2018. https://www.peixebr.com.br/Anuario2018/AnuarioPeixeBR2018.pdf. Accessed 15 October 2019
Pereira BVR, Silva-Zacarin ECM, Costa MJ, Santos ACA, Carmo JB, Nunes B (2019) Cholinesterases characterization of three tropical fish species, and their sensitivity towards specific contaminants. Ecotoxicol Environ Saf 173:482–493. https://doi.org/10.1016/j.ecoenv.2019.01.105
Perkins EJ, Schlenk D (2000) In vivo acetylcholinesterase inhibition, metabolism, and toxicokinetics of aldicarb in channel catfish: role of biotransformation in acute toxicity. Toxicol Sci 53:308–315. https://doi.org/10.1093/toxsci/53.2.308
Pezzementi L, Chatonnet A (2010) Evolution of cholinesterases in the animal kingdom. Chem Biol Interact 187:27–33. https://doi.org/10.1016/j.cbi.2010.03.043
Reis YS, Leite JLR, Almeida CAL, Pereira DSP, Vidal LVO, Araújo FG, Fortes-Silva R (2019) New insights into tambaqui (Colossoma macropomum) feeding behavior and digestive physiology by the self-feeding approach: effects on growth, dial patterns of food digestibility, amylase activity and gastrointestinal transit time. Aquaculture 498:116–122. https://doi.org/10.1016/j.aquaculture.2018.08.054
Salazar-Lugo R, Estrella A, Oliveros A, Rojas-Villarroel E, Villalobos L, Lemus M (2009) Paraquat and temperature affect nonspecific immune response of Colossoma macropomum. Environ Toxicol Pharmacol 27:321–326. https://doi.org/10.1016/j.etap.2008.11.010
Silva KCC, Assis CRD, Oliveira VM, Carvalho LB Jr, Bezerra RS (2013) Kinetic and physicochemical properties of brain acetylcholinesterase from the peacock bass (Cichla ocellaris) and in vitro effect of pesticides and metal ions. Aquat Toxicol 126:191–197. https://doi.org/10.1016/j.aquatox.2012.11.001
Silva ALF, Araújo LD, Gomes LC, Chagas EC (2017) Acute toxicity and sublethal effects of methyl parathion on tambaqui (Colossoma macropomum). J Chem Pharm Res 9:93–97
Sinha AK, Vanparys C, De Boeck G, Kestemont P, Wang N, Nguyen PT, Scippo ML, De Coen W, Robbens J (2010) Expression characteristics of potential biomarker genes in Tra catfish, Pangasianodon hypophthalmus, exposed to trichlorfon. Comp Biochem Physiol Part D Genomics Proteomics 5:207–216. https://doi.org/10.1016/j.cbd.2010.05.001
Soares PRL, Andrade ALC, Santos TPS, Silva SCBL, Silva JF, Santos AR, Souza EHLS, Cunha FM, Teixeira VW, Cadena MRS, Sá FB, Júnior LBC, Cadena PG (2016) Acute and chronic toxicity of the benzoylurea pesticide, lufenuron, in the fish, Colossoma macropomum. Chemosphere 161:412–421. https://doi.org/10.1016/j.chemosphere.2016.07.033
Stewart A, Wu N, Cachat J, Hart P, Gaikwad S, Wong K, Utterback E, Gilder T, Kyzar E, Newman A, Carlos D, Chang K, Hook M, Rhymes C, Caffery M, Greenberg M, Zadina J, Kalueff AV (2011) Pharmacological modulation of anxiety-like phenotypes in adult zebrafish behavioral models. Prog Neuro-Psychopharmacol Biol Psychiatry 35:1421–1431. https://doi.org/10.1016/j.pnpbp.2010.11.035
Varó I, Amat F, Navarro JC (2008) Acute toxicity of dichlorvos to Aphanius iberus (Cuvier & Valenciennes, 1846) and its anti-cholinesterase effects on this species. Aquat Toxicol 88:53–61. https://doi.org/10.1016/j.aquatox.2008.03.004
Venturini FP, Moraes FD, Cortella LRX, Rossi PA, Cruz C, Moraes G (2015) Metabolic effects of trichlorfon (Masoten®) on the neotropical freshwater fish pacu (Piaractus mesopotamicus). Fish Physiol Biochem 41:299–309. https://doi.org/10.1007/s10695-014-9983-y
Yoshimura H, Endoh YS (2005) Acute toxicity to freshwater organisms of antiparasitic drugs for veterinary use. Environ Toxicol 20:60–66. https://doi.org/10.1002/tox.20078
Acknowledgements
This study was partially supported by the ADAPTA project (Adaptações da Biota Aquática da Amazônia)/CNPq/FAPEAM. The authors would like to thank Fabrício Barros de Sousa, Raony César Belém, Maria dos Santos Costa, Rubia Neris Machado, Vinicius Barbosa Costa, Karina Francis de Souza Barbosa, Luana da Silva Nonato, Pedro Vinicius Rodrigues Nunes, and Teresa Cristina for their assistance.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
All experiments were approved by the Ethics Committee for Animal Experimentation (#030/2018 - CEUA/UFAM).
Conflict of interest
The authors declare that they have no conflicts of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Duncan, W.P., Idalino, J.J.S., da Silva, A.G. et al. Acute toxicity of the pesticide trichlorfon and inhibition of acetylcholinesterase in Colossoma macropomum (Characiformes: Serrasalmidae). Aquacult Int 28, 815–830 (2020). https://doi.org/10.1007/s10499-019-00497-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10499-019-00497-w