Paraoxonase activity in sera from Piaractus mesopotamicus Holmberg (Characidae) and Hypostomus punctatus Valenciennes (Siluridae)

Bastos, Vera Lucia F. Cunha; Rossini, Ana; Alves, Marcelo V; Ceccarelli, Paulo S; de Lima, José A. Ferraz; Bastos, Jayme Cunha

doi:10.1590/S0101-81751998000300011

Abstract

A paraoxonase activity present in serum of two Brazilian fish species was consistently assayed at pH 8.5 using 7.5 mM paraoxon final concentration. The paraoxonase activity was more activated by 0.5 M NaCl in serum of Piaractus mesopotanricus Holmberg, 1887 (pacu) than in serum of Hypostomus punctatus Valenciennes, 1840 (cascudo). Apparent values of K M were 3.3 x 10-3 M for cascudo and pacu paraoxonase activity in the presence of 0.5 M NaCl. Apparent maximum velocity values calculated in the presence of 0.5 M NaCl were 6.1 and 6.5 nmole/min/mL of serum for cascudo and pacu, respectively. Vmax/K M ratio values of determinations in the presence and absence of 0.5 M NaCl showed that NaCl had a more evident effect on increasing the affinity of serum paraoxonase for paraoxon in pacu serum. Young specimens of pacu showed a marked decreased paraoxonase serum activity when kept in tanks treated with 0.25 ppm methyl-parathion.

Detoxication; fish; organophosphate; serum esterases

Paraoxonase activity in sera from Piaractus mesopotamicus Holmberg (Characidae) and Hypostomus punctatus Valenciennes (Siluridae)

Vera Lucia F. Cunha Bastos^I; Ana Rossini^I; Marcelo V. Alves^I; Paulo S. Ceccarelli^II; José A. Ferraz de Lima^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

A paraoxonase activity present in serum of two Brazilian fish species was consistently assayed at pH 8.5 using 7.5 mM paraoxon final concentration. The paraoxonase activity was more activated by 0.5 M NaCl in serum of Piaractus mesopotanricus Holmberg, 1887 (pacu) than in serum of Hypostomus punctatus Valenciennes, 1840 (cascudo). Apparent values of K_M were 3.3 x 10^-3 M for cascudo and pacu paraoxonase activity in the presence of 0.5 M NaCl. Apparent maximum velocity values calculated in the presence of 0.5 M NaCl were 6.1 and 6.5 nmole/min/mL of serum for cascudo and pacu, respectively. V_max/K_M ratio values of determinations in the presence and absence of 0.5 M NaCl showed that NaCl had a more evident effect on increasing the affinity of serum paraoxonase for paraoxon in pacu serum. Young specimens of pacu showed a marked decreased paraoxonase serum activity when kept in tanks treated with 0.25 ppm methyl-parathion.

Key words: Detoxication, fish, organophosphate, serum esterases.

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

Recebido em 28.II.1997; aceito em 03.VIII.1998.

ALDRIDGE, W.N. 1953. Serum esterase 2: an enzyme hydrolyzing diethyl p-nitrophenyl phosphate (E600) and its identity with the A-esterase of mammalian sera. Biochem. Jour. 53:117-124.
BENKE, G.M.; K.L. CHEEVER; F.E. MIRER & S.D. MURPHY. 1974. Comparative toxicity, anticholinesterase action and metabolism of methyl parathion and parathion in sunfish and mice. Toxicol. Appl. Pharmacol. 28:97-109.
BRIMBLECOMBE, R.W. 1977. Drugs acting on central cholinergic mechanisms and affecting respiration. Pharcol. Ther. 3:65-74.
BREALEY, C.J.; C.H. WALKER & B.C. BALDWIN. 1980. A-esterase activities in relation to the differential toxicity of pirimiphos-methyl to birds and mammals. Pestic. Sci. 11:546-554.
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. 94C:683-689.
CUNHA BASTOS, V.L.F.; J. CUNHA BASTOS; R.L. MENDONÇA & M.V. CASTRO FARIA. 1988. Main kinetic characteristics of acetylcholinesterase from brain of Hypostomus punctatus, a Brazilian bentonic fish (cascudo). Comp. Biochem. Physiol. 91C:327-331.
DIGGLE, W.M. & J.C. GAGE. 1951. Cholinesterase Inhibition in vitro by OO-Diethyl O-p-nitrophenyl Thiophosphate (Parathion, E 605). Biochem. Jour. 49:491-494.
DUBOIS, K.P. 1961. Potentiation of the toxicity of organophosphorus compounds. Adv. Pest Control Res. 4:117-151.
ECKERSON, H.W.; J. ROMSOM; C. WYTE & B. LA DU. 1983. The human serum paraoxonase polymorphism: Identification of phenotypes by their response to salts. Amer. Jour. Hum. Genet. 35:214-227.
ECOBICHON, D.J. 1996. Toxic effects of pesticides, p. 643-689. In: C.D. KLAASSEN (Ed.). Casarett and Doul's Toxicology - The Basic Science of Poisons. New York, MacGraw-Hill Co. Inc., 5^th ed, 1111p.
FURLONG, C.E.; R.J. RICHTER; S.L. SEIDEL & A.G. MOTULSKY. 1988. Role of genetic polymorphism of human plasma paraoxonase/arylesterase in hydrolysis of the insecticide metabolites chlorpyrifos oxon and paraoxon. Amer. Jour. Hum. Genet. 43:230-238.
FURLONG, C.E.; R.J. RICHTER; S.L. SEIDEL; L.G. COSTA & A.G. MOTULSKY 1989. Spectrophotometric assays for the enzymatic hydrolysis of the active metabolites of chlorpyrifos and parathion by plasma paraoxonase/arylesterase. Anal. Biochem. 180:242-247.
GRAVE, K.; M. ENGLESTAD & N.E. SOLI. 1991a. Utilization of dichlorvos and trichlorfon in salmonid farming in Norway during 1981-1988. Acta Vet. Scand. 32:1-7.
GRAVE, K.; M. ENGLESTAD; 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.
LI, W.F.; C.E. FURLONG & L.G. COSTA. 1993. Serum paraoxonase status: a major factor in determining resistance to organophosphates. Jour. Toxicol. Environ. Health 40:337-346.
______. 1995. Paraoxonase protects against chlorpyrifos toxicity in mice. Toxicol. Letters, 76:219-226.
LINEWEAVER, H. & D. BURKE. 1934. The determination of enzyme dissociation constants. Jour. Amer. Chem. Soc. 56:658-666.
MAIN, A.R. 1976. Structure and inhibitors of cholinesterase, p. 269-353. In: M. GOLDBERG & I. HANIN (Ed.). Biology of Cholinergic Function. New York, Raven Press, 512p.
MURPHY, S.D. 1975. Pesticides, p. 408-453. In: L.J. CASARETT & J. DOULL (Ed.). Toxicology the Basic Science of Poinsons. New York, Macmillan Publishing Co. Inc, 768p.
NATOFF, I.L. 1971. Organophosphorus pesticides. Pharmacol. Prog. Med. Chem. 8:1-37.
NEAL, R.A. 1967. Studies on the metabolism of diethyl 4-nitrophenyl phosphorothionate (parathion) in vitro. Biochem. Jour. 103:183-191.
NEAL, R.A. & K.P. DUBOIS. 1965. Studies on the mechanism of detoxification of cholinergic phosphorothioates. Jour. Pharmacol. Exp. Therapeut. 148 (2):185-192.
PELLIN, M.C.; A. MORETTO; M. LOTTI & E. VILANOVA. 1990. Distribution and some biochemical properties of rat paraoxonase activity. Neurotoxicol. Teratol. 12:611-614.
PLAYFER, J.R.; L.C. EZE; M.F. BULLEN & D.A.P. EVANS. 1976. Genetic polymorphism and interethnic variability of plasma paroxonase activity. Jour. Med. Genet. 13:337-342.
REINER, E.; M.K. JOHNSON & M. JOKANOVIC. 1993. Hydrolysis of some organophosphorus dichlorophenyl esters by hen brain homogenates and rabbit serum compared with hydrolysis of paraoxon. Chem. Biol. Interactions 87:127-131.
WALKER, C.H. 1993. The classification of esterases which hydrolyse organophosphates: recent developments. Chem. Biol. Interactions 87:17-24.
WALKER, C.H. & M.I. MACKNESS. 1987. A-esterases and their role in regulating the toxicity of organophosphates. Arch. Toxicol. 60:30-33.
ZECH, R. & K. ZÜRCHER. 1974. Organophosphate splitting serum enzymes in different mammals. Comp. Biochem. Physiol. 48B:427-433.

Publication Dates

Publication in this collection
10 July 2009
Date of issue
1998

History

Accepted
03 Aug 1998
Received
28 Feb 1997

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

[1] ALDRIDGE, W.N. 1953. Serum esterase 2: an enzyme hydrolyzing diethyl p-nitrophenyl phosphate (E600) and its identity with the A-esterase of mammalian sera. Biochem. Jour. 53:117-124.

[2] BENKE, G.M.; K.L. CHEEVER; F.E. MIRER & S.D. MURPHY. 1974. Comparative toxicity, anticholinesterase action and metabolism of methyl parathion and parathion in sunfish and mice. Toxicol. Appl. Pharmacol. 28:97-109.

[3] BRIMBLECOMBE, R.W. 1977. Drugs acting on central cholinergic mechanisms and affecting respiration. Pharcol. Ther. 3:65-74.

[4] BREALEY, C.J.; C.H. WALKER & B.C. BALDWIN. 1980. A-esterase activities in relation to the differential toxicity of pirimiphos-methyl to birds and mammals. Pestic. Sci. 11:546-554.

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

[6] 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. 94C:683-689.

[7] CUNHA BASTOS, V.L.F.; J. CUNHA BASTOS; R.L. MENDONÇA & M.V. CASTRO FARIA. 1988. Main kinetic characteristics of acetylcholinesterase from brain of Hypostomus punctatus, a Brazilian bentonic fish (cascudo). Comp. Biochem. Physiol. 91C:327-331.

[8] DIGGLE, W.M. & J.C. GAGE. 1951. Cholinesterase Inhibition in vitro by OO-Diethyl O-p-nitrophenyl Thiophosphate (Parathion, E 605). Biochem. Jour. 49:491-494.

[9] DUBOIS, K.P. 1961. Potentiation of the toxicity of organophosphorus compounds. Adv. Pest Control Res. 4:117-151.

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

[11] ECOBICHON, D.J. 1996. Toxic effects of pesticides, p. 643-689. In: C.D. KLAASSEN (Ed.). Casarett and Doul's Toxicology - The Basic Science of Poisons. New York, MacGraw-Hill Co. Inc., 5^th ed, 1111p.

[12] FURLONG, C.E.; R.J. RICHTER; S.L. SEIDEL & A.G. MOTULSKY. 1988. Role of genetic polymorphism of human plasma paraoxonase/arylesterase in hydrolysis of the insecticide metabolites chlorpyrifos oxon and paraoxon. Amer. Jour. Hum. Genet. 43:230-238.

[13] FURLONG, C.E.; R.J. RICHTER; S.L. SEIDEL; L.G. COSTA & A.G. MOTULSKY 1989. Spectrophotometric assays for the enzymatic hydrolysis of the active metabolites of chlorpyrifos and parathion by plasma paraoxonase/arylesterase. Anal. Biochem. 180:242-247.

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

[15] GRAVE, K.; M. ENGLESTAD; 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.

[16] LI, W.F.; C.E. FURLONG & L.G. COSTA. 1993. Serum paraoxonase status: a major factor in determining resistance to organophosphates. Jour. Toxicol. Environ. Health 40:337-346.

[17] ______. 1995. Paraoxonase protects against chlorpyrifos toxicity in mice. Toxicol. Letters, 76:219-226.

[18] LINEWEAVER, H. & D. BURKE. 1934. The determination of enzyme dissociation constants. Jour. Amer. Chem. Soc. 56:658-666.

[19] MAIN, A.R. 1976. Structure and inhibitors of cholinesterase, p. 269-353. In: M. GOLDBERG & I. HANIN (Ed.). Biology of Cholinergic Function. New York, Raven Press, 512p.

[20] MURPHY, S.D. 1975. Pesticides, p. 408-453. In: L.J. CASARETT & J. DOULL (Ed.). Toxicology the Basic Science of Poinsons. New York, Macmillan Publishing Co. Inc, 768p.

[21] NATOFF, I.L. 1971. Organophosphorus pesticides. Pharmacol. Prog. Med. Chem. 8:1-37.

[22] NEAL, R.A. 1967. Studies on the metabolism of diethyl 4-nitrophenyl phosphorothionate (parathion) in vitro. Biochem. Jour. 103:183-191.

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

[24] PELLIN, M.C.; A. MORETTO; M. LOTTI & E. VILANOVA. 1990. Distribution and some biochemical properties of rat paraoxonase activity. Neurotoxicol. Teratol. 12:611-614.

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

[26] REINER, E.; M.K. JOHNSON & M. JOKANOVIC. 1993. Hydrolysis of some organophosphorus dichlorophenyl esters by hen brain homogenates and rabbit serum compared with hydrolysis of paraoxon. Chem. Biol. Interactions 87:127-131.

[27] WALKER, C.H. 1993. The classification of esterases which hydrolyse organophosphates: recent developments. Chem. Biol. Interactions 87:17-24.

[28] WALKER, C.H. & M.I. MACKNESS. 1987. A-esterases and their role in regulating the toxicity of organophosphates. Arch. Toxicol. 60:30-33.

[29] ZECH, R. & K. ZÜRCHER. 1974. Organophosphate splitting serum enzymes in different mammals. Comp. Biochem. Physiol. 48B:427-433.