Paraoxonase activity in liver of Pacu, Piaractus mesopotamicus Holmberg (Characidae)

Bastos, Vera Lucia F. Cunha; Folly, Evelize; Rossini, Ana; Ceccarelli, Paulo S.; Senhorini, José A.; Bastos, Jayme Cunha

doi:10.1590/S0101-81751998000300012

Abstract

Enzymatic production of p-nitrophenol in liver of Piaractus mesopotamicus Holmberg, 1887 was consistently assayed at pH 8.5 using 7.5 mM paraoxon as substrate. This pacu liver paraoxonase activity was activated by NaCl. Apparent values of K M were 2.42 x 10-3 M in the presence of 0.5 M NaCl and 8.99 x 10-3 M without NaCl. Apparent maximum velocity values calculated in the absence and presence of 0.5 M NaCl were 1.09 x 10-3 µmoles/min/mg of proteins and 1.29 x 100-3 µmoles/min/mg of proteins, respectively. These Vmax values are fifty-fold the value described for trout (Salmo trutta Linnaeus, 1758) liver paraoxonase. Paraoxonase activity of pacu liver homogenates was recovered as much in cytosolic as in particulate cellular subtractions, but the particulate subtractions showed higher specific activities. The data presented here indicate that hepatic hydrolysis of organophosphorous pesticides may not be an important detoxification process in pacu.

Detoxication; esterase; fish; organophosphate

Paraoxonase activity in liver of Pacu, Piaractus mesopotamicus Holmberg (Characidae)

Vera Lucia F. Cunha Bastos^I; Evelize Folly^I; Ana Rossini^I; Paulo S. Ceccarelli^II; José A. Senhorini^II; Jayme Cunha Bastos^I

^IDepartamento de Bioquímica, Instituto de Biologia Roberto Alcântara Gomes, Universidade Estadual do Rio de Janeiro. Avenida Prof. Manuel de Abreu 48, 20550-170 Rio de Janeiro, Rio de Janeiro, Brasil

^IICentro Nacional de Pesquisas em Peixes Tropicais. Caixa Postal 64, 13630-970 Pirassununga, São Paulo, Brasil

ABSTRACT

Enzymatic production of p-nitrophenol in liver of Piaractus mesopotamicus Holmberg, 1887 was consistently assayed at pH 8.5 using 7.5 mM paraoxon as substrate. This pacu liver paraoxonase activity was activated by NaCl. Apparent values of K_M were 2.42 x 10^-3 M in the presence of 0.5 M NaCl and 8.99 x 10^-3 M without NaCl. Apparent maximum velocity values calculated in the absence and presence of 0.5 M NaCl were 1.09 x 10^-3µmoles/min/mg of proteins and 1.29 x 100^-3µmoles/min/mg of proteins, respectively. These V_max values are fifty-fold the value described for trout (Salmo trutta Linnaeus, 1758) liver paraoxonase. Paraoxonase activity of pacu liver homogenates was recovered as much in cytosolic as in particulate cellular subtractions, but the particulate subtractions showed higher specific activities. The data presented here indicate that hepatic hydrolysis of organophosphorous pesticides may not be an important detoxification process in pacu.

Key words: Detoxication, esterase, fish, organophosphate

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ACKNOWLEDGEMENTS. The authors are grateful to the support provided by the Rio de Janeiro State Research Support Foundation (FAPERJ).

REFERENCES

BURNS, K.A. 1976. Microsomal mixed function oxidases in an estuarine fish, Fundulus heteroclitus, and their induction as a result of environmental contamination. Comp. Biochem. Physiol. 53B:443-446.

CHEMNITIUS, J-M.; H. LOSCH; K. LOSCH & R. ZECH. 1983. Organophosphate detoxicating hydrolases in different vertebrate species. Comp. Biochem. Physiol., 76C:85-93.

CUNHA BASTOS, J.; V.L.F. CUNHA BASTOS; A. ROSSINI; H. FORTINI & M.V. CASTRO FARIA. 1992. Activation of parathion by liver of Hypostomus punctatus, a Brazilian benthic fish (Cascudo). Comp. Biochem. Physiol. 102C:561-566.

CUNHA BASTOS, V.L.F.; J. CUNHA BASTOS; A. ROSSINI; M.V. ALVES; P.S. CECCARELLI & J.A. FERRAZ DE LIMA. Paraoxon esterase activity in sera from Piaractus mesopotamicus Holmberg (Characidae) and Hypostomus punctatus Valenciennes (Siluridae). Revta bras. Zool. 15.

ECKERSON, H.W.; J. ROMSON; C. WYTE & B.N. LA DU. 1983. The human serum paraoxonase polymorphism: identification of phenotypes by their response to salts. Amer. Jour. Hum. Genet. 35:214-227.

ERDÖS, E.G.; C.R. DEBAY & M.P. WESTERMAN. 1959. Activation and inhibition of the arylesterase of human serum. Nature 184:430-431.

FARIA, H.C. & A. BRASIL. 1959. Adaptação foto-colorimétrica do método de Greenberg, para dosagem das proteínas do soro. O Hospital 55:597-601.

GAINES, T.; W.J. HAYES & R.E. LINDER. 1966. Liver metabolism of anticholinesterase compounds in rats: Relation to toxicity. Nature 209:88-89.

GELDMACHER-VON MALLINCKRODT, M. & T.L. DIEBGEN. 1988. The human serum paraoxonase - polymorphism and specificity. Toxicol. Environ. Chem. 18:79-196.

GRAVE, K.; M. ENGELSTAD & N.E SOLLY. 1991a. Utilization of dichlorvos and trichlorfon in salmonid farming in Norvvay during 1981-1988. 1991a. Acta Vet. Scand. 32:1-7.

GRAVE, K.; M. ENGELSTAD; N.E. SOLI & E.L. TOVERUD. 1991b. Clinical use of dichlorvos (Nuvan) and trichlorfon (Neguvon) in the treatment of salmon louse, Lepeophtheirus salmonis. compliance with the recommended treatment procedures. Acta Vet. Scand. 32:9-14.

GREENBERG, D.M. 1929. The colorimetric determination of the serum proteins. Jour. Biol. Chem. 82:545-550.

HINTON, D.E.; J.E. KLAUNIG & M.M. LIPSKY. 1978. PCB-induced alterations in teleost liver: A model for environmental disease in fish. Mar. Fish Rev. 40:47-50.

HINTON, D.E.; J. A. HAMPTON & R.C. LANTZ. 1985. Morphometric analysis of liver in rainbow trout quantitatively defining an organ of xenobiotic metabolism. Mar. Environ. Res. 17:238-239.

LI, W.F.; C.E. FURLONG & L.G. COSTA. 1995. Paraoxonase protects against chlorpyrifos toxicity in mice. Toxicol, letters 76:219-226.

NEAL, R.A. 1967. Studies on the metabolism of diethyl-4-nitrophenyl phosphorothionate (parathion) in vitro. Biochem. Jour. 108:183-191.

NEAL, R.A. & K.P. DUBOIS. 1965. Studies on the mechanism of detoxification of cholinergic phosphorothioates. Jour. Pharmacol. Exp. Therapeut. 148 (2):185-192.

NOWAK, B. 1996. Relationship between endosulfan residue level and ultrastructural changes in the liver of Catfish, Tandanus tandanus. Arch. Environ. Contain. Toxicol. 30:195-202.

PEDERSEN, M.G.; W.K. HERSHBERGER; P.K. ZACHARIAH & M.R. JUCHAU. 1976. Hepatic biotransformation of environmental xenobiotics in six strains of rainbow trout (Salmo gairdneri). Jour. Fish. Res. Board Can. 33:666-675.

PLA, A. & M.K. JOHNSON. 1989. Degradation by rat tissues in vitro of organophosphorus esters which inhibit Cholinesterase. Biochem. Pharmacol. 38:1527-1533.

PLAYFER, J.R.; L.C. EZE; M.F. BULLEN & D.A.P. EVANS. 1976. Genetic polymorphism and interethnic variability of plasma paraoxonase activity. Jour. Med. Genet. 13:337-342.

POND, A.L.; H.W. CHAMBERS & J.E. CHAMBERS. 1995. Organophosphate detoxication potential of various rat tissues via A-esterase and aliesterase activities. Toxicol. Letters 78:245-252.

SMOLEN, A.; H.W. ECKERSON; K.N. GAN; N. HAILAT & B.N. LA DU. 1991. Characteristics of the genetically determined allozymic forms of human serum paraoxonase/arylesterase. Drug Metab. Dispos. 19:107-112.

SULTATOS, L.G.; L.D. MINOR & S.D. MURPHY. 1985. Metabolic activation of phosphorothioate pesticides: role of the liver. Jour. Pharmacol. Exp. Therapeut. 232:624-628.

Recebido em 24.III.1997; aceito em 03.VIII.1998.

BURNS, K.A. 1976. Microsomal mixed function oxidases in an estuarine fish, Fundulus heteroclitus, and their induction as a result of environmental contamination. Comp. Biochem. Physiol. 53B:443-446.
CHEMNITIUS, J-M.; H. LOSCH; K. LOSCH & R. ZECH. 1983. Organophosphate detoxicating hydrolases in different vertebrate species. Comp. Biochem. Physiol., 76C:85-93.
CUNHA BASTOS, J.; V.L.F. CUNHA BASTOS; A. ROSSINI; H. FORTINI & M.V. CASTRO FARIA. 1992. Activation of parathion by liver of Hypostomus punctatus, a Brazilian benthic fish (Cascudo). Comp. Biochem. Physiol. 102C:561-566.
CUNHA BASTOS, V.L.F.; J. CUNHA BASTOS; A. ROSSINI; M.V. ALVES; P.S. CECCARELLI & J.A. FERRAZ DE LIMA. Paraoxon esterase activity in sera from Piaractus mesopotamicus Holmberg (Characidae) and Hypostomus punctatus Valenciennes (Siluridae). Revta bras. Zool. 15.
ECKERSON, H.W.; J. ROMSON; C. WYTE & B.N. LA DU. 1983. The human serum paraoxonase polymorphism: identification of phenotypes by their response to salts. Amer. Jour. Hum. Genet. 35:214-227.
ERDÖS, E.G.; C.R. DEBAY & M.P. WESTERMAN. 1959. Activation and inhibition of the arylesterase of human serum. Nature 184:430-431.
FARIA, H.C. & A. BRASIL. 1959. Adaptação foto-colorimétrica do método de Greenberg, para dosagem das proteínas do soro. O Hospital 55:597-601.
GAINES, T.; W.J. HAYES & R.E. LINDER. 1966. Liver metabolism of anticholinesterase compounds in rats: Relation to toxicity. Nature 209:88-89.
GELDMACHER-VON MALLINCKRODT, M. & T.L. DIEBGEN. 1988. The human serum paraoxonase - polymorphism and specificity. Toxicol. Environ. Chem. 18:79-196.
GRAVE, K.; M. ENGELSTAD & N.E SOLLY. 1991a. Utilization of dichlorvos and trichlorfon in salmonid farming in Norvvay during 1981-1988. 1991a. Acta Vet. Scand. 32:1-7.
GRAVE, K.; M. ENGELSTAD; N.E. SOLI & E.L. TOVERUD. 1991b. Clinical use of dichlorvos (Nuvan) and trichlorfon (Neguvon) in the treatment of salmon louse, Lepeophtheirus salmonis. compliance with the recommended treatment procedures. Acta Vet. Scand. 32:9-14.
GREENBERG, D.M. 1929. The colorimetric determination of the serum proteins. Jour. Biol. Chem. 82:545-550.
HINTON, D.E.; J.E. KLAUNIG & M.M. LIPSKY. 1978. PCB-induced alterations in teleost liver: A model for environmental disease in fish. Mar. Fish Rev. 40:47-50.
HINTON, D.E.; J. A. HAMPTON & R.C. LANTZ. 1985. Morphometric analysis of liver in rainbow trout quantitatively defining an organ of xenobiotic metabolism. Mar. Environ. Res. 17:238-239.
LI, W.F.; C.E. FURLONG & L.G. COSTA. 1995. Paraoxonase protects against chlorpyrifos toxicity in mice. Toxicol, letters 76:219-226.
NEAL, R.A. 1967. Studies on the metabolism of diethyl-4-nitrophenyl phosphorothionate (parathion) in vitro. Biochem. Jour. 108:183-191.
NEAL, R.A. & K.P. DUBOIS. 1965. Studies on the mechanism of detoxification of cholinergic phosphorothioates. Jour. Pharmacol. Exp. Therapeut. 148 (2):185-192.
NOWAK, B. 1996. Relationship between endosulfan residue level and ultrastructural changes in the liver of Catfish, Tandanus tandanus. Arch. Environ. Contain. Toxicol. 30:195-202.
PEDERSEN, M.G.; W.K. HERSHBERGER; P.K. ZACHARIAH & M.R. JUCHAU. 1976. Hepatic biotransformation of environmental xenobiotics in six strains of rainbow trout (Salmo gairdneri). Jour. Fish. Res. Board Can. 33:666-675.
PLA, A. & M.K. JOHNSON. 1989. Degradation by rat tissues in vitro of organophosphorus esters which inhibit Cholinesterase. Biochem. Pharmacol. 38:1527-1533.
PLAYFER, J.R.; L.C. EZE; M.F. BULLEN & D.A.P. EVANS. 1976. Genetic polymorphism and interethnic variability of plasma paraoxonase activity. Jour. Med. Genet. 13:337-342.
POND, A.L.; H.W. CHAMBERS & J.E. CHAMBERS. 1995. Organophosphate detoxication potential of various rat tissues via A-esterase and aliesterase activities. Toxicol. Letters 78:245-252.
SMOLEN, A.; H.W. ECKERSON; K.N. GAN; N. HAILAT & B.N. LA DU. 1991. Characteristics of the genetically determined allozymic forms of human serum paraoxonase/arylesterase. Drug Metab. Dispos. 19:107-112.
SULTATOS, L.G.; L.D. MINOR & S.D. MURPHY. 1985. Metabolic activation of phosphorothioate pesticides: role of the liver. Jour. Pharmacol. Exp. Therapeut. 232:624-628.

Publication Dates

Publication in this collection
10 July 2009
Date of issue
1998

History

Accepted
03 Aug 1998
Received
24 Mar 1997

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] BURNS, K.A. 1976. Microsomal mixed function oxidases in an estuarine fish, Fundulus heteroclitus, and their induction as a result of environmental contamination. Comp. Biochem. Physiol. 53B:443-446.

[2] CHEMNITIUS, J-M.; H. LOSCH; K. LOSCH & R. ZECH. 1983. Organophosphate detoxicating hydrolases in different vertebrate species. Comp. Biochem. Physiol., 76C:85-93.

[3] CUNHA BASTOS, J.; V.L.F. CUNHA BASTOS; A. ROSSINI; H. FORTINI & M.V. CASTRO FARIA. 1992. Activation of parathion by liver of Hypostomus punctatus, a Brazilian benthic fish (Cascudo). Comp. Biochem. Physiol. 102C:561-566.

[4] CUNHA BASTOS, V.L.F.; J. CUNHA BASTOS; A. ROSSINI; M.V. ALVES; P.S. CECCARELLI & J.A. FERRAZ DE LIMA. Paraoxon esterase activity in sera from Piaractus mesopotamicus Holmberg (Characidae) and Hypostomus punctatus Valenciennes (Siluridae). Revta bras. Zool. 15.

[5] ECKERSON, H.W.; J. ROMSON; C. WYTE & B.N. LA DU. 1983. The human serum paraoxonase polymorphism: identification of phenotypes by their response to salts. Amer. Jour. Hum. Genet. 35:214-227.

[6] ERDÖS, E.G.; C.R. DEBAY & M.P. WESTERMAN. 1959. Activation and inhibition of the arylesterase of human serum. Nature 184:430-431.

[7] FARIA, H.C. & A. BRASIL. 1959. Adaptação foto-colorimétrica do método de Greenberg, para dosagem das proteínas do soro. O Hospital 55:597-601.

[8] GAINES, T.; W.J. HAYES & R.E. LINDER. 1966. Liver metabolism of anticholinesterase compounds in rats: Relation to toxicity. Nature 209:88-89.

[9] GELDMACHER-VON MALLINCKRODT, M. & T.L. DIEBGEN. 1988. The human serum paraoxonase - polymorphism and specificity. Toxicol. Environ. Chem. 18:79-196.

[10] GRAVE, K.; M. ENGELSTAD & N.E SOLLY. 1991a. Utilization of dichlorvos and trichlorfon in salmonid farming in Norvvay during 1981-1988. 1991a. Acta Vet. Scand. 32:1-7.

[11] GRAVE, K.; M. ENGELSTAD; N.E. SOLI & E.L. TOVERUD. 1991b. Clinical use of dichlorvos (Nuvan) and trichlorfon (Neguvon) in the treatment of salmon louse, Lepeophtheirus salmonis. compliance with the recommended treatment procedures. Acta Vet. Scand. 32:9-14.

[12] GREENBERG, D.M. 1929. The colorimetric determination of the serum proteins. Jour. Biol. Chem. 82:545-550.

[13] HINTON, D.E.; J.E. KLAUNIG & M.M. LIPSKY. 1978. PCB-induced alterations in teleost liver: A model for environmental disease in fish. Mar. Fish Rev. 40:47-50.

[14] HINTON, D.E.; J. A. HAMPTON & R.C. LANTZ. 1985. Morphometric analysis of liver in rainbow trout quantitatively defining an organ of xenobiotic metabolism. Mar. Environ. Res. 17:238-239.

[15] LI, W.F.; C.E. FURLONG & L.G. COSTA. 1995. Paraoxonase protects against chlorpyrifos toxicity in mice. Toxicol, letters 76:219-226.

[16] NEAL, R.A. 1967. Studies on the metabolism of diethyl-4-nitrophenyl phosphorothionate (parathion) in vitro. Biochem. Jour. 108:183-191.

[17] NEAL, R.A. & K.P. DUBOIS. 1965. Studies on the mechanism of detoxification of cholinergic phosphorothioates. Jour. Pharmacol. Exp. Therapeut. 148 (2):185-192.

[18] NOWAK, B. 1996. Relationship between endosulfan residue level and ultrastructural changes in the liver of Catfish, Tandanus tandanus. Arch. Environ. Contain. Toxicol. 30:195-202.

[19] PEDERSEN, M.G.; W.K. HERSHBERGER; P.K. ZACHARIAH & M.R. JUCHAU. 1976. Hepatic biotransformation of environmental xenobiotics in six strains of rainbow trout (Salmo gairdneri). Jour. Fish. Res. Board Can. 33:666-675.

[20] PLA, A. & M.K. JOHNSON. 1989. Degradation by rat tissues in vitro of organophosphorus esters which inhibit Cholinesterase. Biochem. Pharmacol. 38:1527-1533.

[21] PLAYFER, J.R.; L.C. EZE; M.F. BULLEN & D.A.P. EVANS. 1976. Genetic polymorphism and interethnic variability of plasma paraoxonase activity. Jour. Med. Genet. 13:337-342.

[22] POND, A.L.; H.W. CHAMBERS & J.E. CHAMBERS. 1995. Organophosphate detoxication potential of various rat tissues via A-esterase and aliesterase activities. Toxicol. Letters 78:245-252.

[23] SMOLEN, A.; H.W. ECKERSON; K.N. GAN; N. HAILAT & B.N. LA DU. 1991. Characteristics of the genetically determined allozymic forms of human serum paraoxonase/arylesterase. Drug Metab. Dispos. 19:107-112.

[24] SULTATOS, L.G.; L.D. MINOR & S.D. MURPHY. 1985. Metabolic activation of phosphorothioate pesticides: role of the liver. Jour. Pharmacol. Exp. Therapeut. 232:624-628.