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Characterization and in vitro sensitivity of cholinesterases of gilthead seabream (Sparus aurata) to organophosphate pesticides

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Abstract

The characterization of cholinesterase activity in brain and muscle of gilthead seabream was carried out using four specific substrates and three selective inhibitors. In addition, K m and V max were calculated from the Michaelis–Menten equation for ASCh and BSCh substrates. Finally, the in vitro sensitivity of brain and muscle cholinesterases to three organophosphates (OPs) was also investigated by estimating inhibition kinetics. The results indicate that AChE is the enzyme present in the brain, whereas in muscle, a typical AChE form is present along with an atypical form of BChE. Very low ChE activity was found in plasma with all substrates used. The inhibitory potency of the studied OPs on brain and muscle AChEs based on bimolecular inhibition constants (k i ) was: omethoate < dichlorvos < azinphosmethyl-oxon. Furthermore, muscle BChE was found to be several orders of magnitude (from 2 to 4) more sensitive than brain and muscle AChE inhibition by dichlorvos and omethoate.

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References

  • Alpuche-Gual L, Gold-Bouchot G (2008) Determination of esterase activity and characterization of cholinesterases in the reef fish Haemulon plumier. Ecotoxicol Environ Saf 71:787–797. doi:10.1016/j.ecoenv.2008.01.024

    Article  Google Scholar 

  • Arufe MI, Arellano JM, García L, Albendín G, Sarasquete C (2007) Cholinesterase activity in gilthead seabream (Sparus aurata) larvae: characterization and sensitivity to the organophosphate azinphosmethyl. Aquat Toxicol 84:328–336. doi:10.1016/j.aquatox.2007.06.009

    Article  CAS  PubMed  Google Scholar 

  • Assis CRD, Bezerra RS, Carvalho LB Jr (2011) Fish cholinesterases as biomarkers of organophosphorus and carbamate pesticides. In: Stoytcheva M (ed) Pesticides in the modern world - Pests control and pesticides exposure and toxicity assessment. InTech, pp 253–278. doi:10.5772/20777

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

    Article  CAS  PubMed  Google Scholar 

  • Carr RL, Chambers JE (1996) Kinetic analysis of the in vitro inhibition, aging, and reactivation of brain acetylcholinesterase from rat and channel catfish by paraoxon and chlorpyrifos-oxon. Toxicol Appl Pharmacol 139:365–373. doi:10.1006/taap.1996.0177

    Article  CAS  PubMed  Google Scholar 

  • Chuiko GM (2000) Comparative study of acetylcholinesterase and butyrylcholinesterase in brain and serum of several freshwater fish: specific activities and in vitro inhibition by DDVP, and organophosphorus pesticide. Comp Biochem Physiol 127C:233–242. doi:10.1016/S0742-8413(00)00150-X

    CAS  Google Scholar 

  • Chuiko GM, Podgornaya VA, Zhelnin YY (2003) Acetylcholinesterase and butyrylcholinesterase activities in brain and plasma of freshwater teleosts: cross-species and cross-family differences. Comp Biochem Physiol 135B:55–61. doi:10.1016/S1096-4959(03)00048-4

    Article  CAS  Google Scholar 

  • De la Torre FR, Ferrari L, Salibián A (2002) Freshwater pollution biomarker: response of brain acetylcholinesterase activity in two fish species. Comp Biochem Physiol 131C:271–280. doi:10.1016/S1532-0456(02)00014-5

    Google Scholar 

  • Ellman GL, Courtney KD, Andres V, FeatherstoneI I, Featherstone RM (1961) A new and rapid colometric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95. doi:10.1016/0006-2952(61)90145-9

    Article  CAS  PubMed  Google Scholar 

  • Guilhermino L, Lopes MC, Carvalho AP, Soares AMVM (1996) Inhibition of acetylcholinesterase activity as effect criterion in acute tests with juvenile Daphnia magna. Chemosphere 32:727–738. doi:10.1016/0045-6535(95)00360-6

    Article  CAS  PubMed  Google Scholar 

  • Herzsprung P, Weil L, Niessner R (1992) Measurement of bimolecular rate constants k i of the cholinesterase inactivation reaction by 55 insecticides and of the influence of various pyridiniumoximes on k i . Int J Environ Anal Chem 47:181–200

    Article  CAS  Google Scholar 

  • Johnson JA, Wallace KB (1987) Species-related differences in the inhibition of brain acetylcholinesterase by paraoxon and malaoxon. Toxicol Appl Pharmacol 88:234–241. doi:10.1016/0041-008X(87)90009-3

    Article  CAS  PubMed  Google Scholar 

  • Jung JH, Addison RF, Shim WJ (2007) Characterization of cholinesterases in marbled sole, Limanda yokohamae, and their inhibition in vitro by the fungicide iprobenfos. Mar Environ Res 63:471–478. doi:10.1016/j.marenvres.2006.12.007

    Article  CAS  PubMed  Google Scholar 

  • Kamrin MA (2000) Pesticides profiles. Toxicology, environmental impact, and fate. CRC Press LLC, CRC Press LLC

    Google Scholar 

  • Kemp RJ, Wallace KB (1990) Molecular determinants of species-selective inhibition of brain acetylcholinesterase. Toxicol Appl Pharmacol 104:246–258. doi:10.1016/0041-008X(90)90299-A

    Article  CAS  PubMed  Google Scholar 

  • Kitz R, Wilson IB (1962) Ester of methanesulfonic acid as irreversible inhibitors of acetylcholinesterase. J Biol Chem 237:3245–3249

    CAS  PubMed  Google Scholar 

  • Main AR, Iverson F (1966) Measurement of the affinity and phosphorylation constants governing irreversible inhibition of cholinesterases by di-isopropyl phosphofluoridate. Biochem J 100:525–531

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Oliveira MM, Filho MVS, Bastos VLF, Fernandes FC, Bastos JC (2007) Brain acetylcholinesterase as a marine pesticide biomarker using brazilian fishes. Mar Environ Res 63:303–312. doi:10.1016/j.marenvres.2006.10.002

    Article  CAS  PubMed  Google Scholar 

  • Quintaneiro C, Monteiro M, Pastorinho R, Soares AMVM, Noqueira AJA, Morgado F, Guilhermino L (2006) Environmental pollution and natural populations: a biomarkers case study from the Iberian Atlantic Coast. Mar Pollut Bull 52:1406–1413. doi:10.1016/j.marpolbul.2006.04.002

    Article  CAS  PubMed  Google Scholar 

  • Rendon Von Osten J, Ortíz-Arana A, Guilhermino L, Soares AMVM (2005) In vivo evaluation of three biomarkers in the mosquitofish (Gambusia yucatana) exposed to pesticides. Chemosphere 58:627–636. doi:10.1016/j.chemosphere.2004.08.065

    Article  CAS  PubMed  Google Scholar 

  • Rodríguez-Fuentes G, Gold-Bouchot G (2004) Characterization of cholinesterase activity from different tissues of Nile tilapia (Oreochromis niloticus). Mar Environ Res 58:505–509. doi:10.1016/j.marenvres.2004.03.037

    Article  PubMed  Google Scholar 

  • Salles JB, Cunha Bastos VLF, Silva Filho MV, Machado OLT, Salles CMC, Giovanni de Simone S, Cunha Bastos J (2006) A novel butyrylcholinesterase from serum of Leporinus macrocephalus, a neotropical fish. Biochimie 88:59–68. doi:10.1016/j.biochi.2005.06.017

    Article  CAS  PubMed  Google Scholar 

  • Shaonan I, Xianchuan X, Guaonian Z, Yajun T (2004) Kinetic characters and resistance to inhibition of crude and purified brain acetylcholinesterase of three freshwater fishes by organophosphates. Aquat Toxicol 68:293–299. doi:10.1016/j.aquatox.2004.03.013

    Article  PubMed  Google Scholar 

  • Silva Filho MVS, Oliveira MM, Salles JB, Cunha Bastos VLF, Cassano VPF, Cunha Bastos J (2004) Methyl-paraoxon comparative inhibition kinetics for acetylthiocholinesterases from brain of neotropical fishes. Toxicol Lett 153:247–254. doi:10.1016/j.toxlet.2004.04.026

    Article  CAS  PubMed  Google Scholar 

  • Solè M, Vega S, Varò I (2012) Characterization of type “B” esterases and hepatic CYP450 isoenzimes in Senegalese sole for their further application in monitoring studies. Ecotoxicol Environ Saf 78:72–79. doi:10.1016/j.ecoenv.2011.11.013

    Article  PubMed  Google Scholar 

  • Sturm A, da Silva de Assis HC, Hansen PD (1999) Cholinesterases of marine teleost fish: enzymological characterization and potential use in the monitoring of neurotoxic contamination. Mar Environ Res 47:389–398. Correction: Mar Environ Res 49(2000):95. doi:10.1016/S0141-1136(98)00127-5

  • Sturm A, Wogran J, Segner H, Liess M (2000) Different sensitivity to organophosphates of acetylcholinesterase and butyrylcholinesterase from three-spined stickeback (Gasterosteus aculeatus): application in biomonitoring. Environ Toxicol Chem 19:1607–1615. doi:10.1002/etc.5620190618

    Article  CAS  Google Scholar 

  • Tortelli V, Colares EP, Robaldo RB, Nery LEM, Pinho GLL, Bianchini A, Monserrat JM (2006) Importance of cholinesterase kinetic parameters in environmental monitoring using estuarine fish. Chemosphere 65:560–566. doi:10.1016/j.chemosphere.2006.02.047

    Article  CAS  PubMed  Google Scholar 

  • Varò I, Navarro JC, Nunes B, Guilhermino L (2007) Effects of dichlorvos aquaculture treatments on selected biomarkers of gilthead seabream (Sparus aurata L.) fingerlings. Aquaculture 266:87–96. doi:10.1016/j.aquaculture.2007.02.045

    Article  Google Scholar 

  • Wang C, Murphy SD (1982) Kinetic analysis of species difference in acetylcholinesterase sensitivity to organophosphate insecticides. Toxicol Appl Pharmacol 66:409–419. doi:10.1016/0041-008X(82)90307-6

    Article  CAS  PubMed  Google Scholar 

  • Wilson BW (2010) Cholinesterases. Hayes’s handbook of pesticide toxicology. Academic Press, New York, pp 1457–1478

    Chapter  Google Scholar 

Download references

Acknowledgments

The authors are grateful to the Laboratory of Marine Culture at University of Cádiz for providing the experimental fish used in this study. This work was supported by Plan Nacional de I + D, Sciences and Technology Minister (Project: CTM2004-05718) and in part by Plan Andaluz de Investigación (PAI group: RNM-345).

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Authors and Affiliations

  1. Toxicology Laboratory, Faculty of Marine and Environmental Sciences, University of Cádiz, Polígono Río San Pedro s/n, 11510, Puerto Real, Cádiz, Spain

    G. Albendín, J. M. Arellano & M. I. Arufe

  2. Analytical Chemical Department, Faculty of Marine and Environmental Sciences, University of Cádiz, Polígono Río San Pedro s/n, 11510, Puerto Real, Cádiz, Spain

    M. P. Mánuel-Vez

  3. Institute of Marine Sciences of Andalucía (ICMAN-CSIC), Avda. República Saharaui s/n, 11510, Puerto Real, Cádiz, Spain

    C. Sarasquete

Authors
  1. G. Albendín
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  2. J. M. Arellano
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  3. M. P. Mánuel-Vez
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  4. C. Sarasquete
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  5. M. I. Arufe
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Corresponding author

Correspondence to G. Albendín.

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Albendín, G., Arellano, J.M., Mánuel-Vez, M.P. et al. Characterization and in vitro sensitivity of cholinesterases of gilthead seabream (Sparus aurata) to organophosphate pesticides. Fish Physiol Biochem 43, 455–464 (2017). https://doi.org/10.1007/s10695-016-0299-y

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  • DOI: https://doi.org/10.1007/s10695-016-0299-y

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