Intraspecific variation in the venom electrophoretic profile of recently captured Crotalus durissus terrificus (Laurenti, 1768) snakes

MAGRO, A. J.; SILVA, R. J. DA; RAMOS, P. R. R.; CHERUBINI, A. L.; HATAYDE, M. R.

doi:10.1590/S0104-79302001000200010

Abstract

The aim of this study was to compare individual venom samples of Crotalus durissus terrificus recently captured in the wild to evaluate possible differences in venom protein composition. Protein levels were quantified by biochemical method (Biuret) and then submitted to electrophoresis. Electrophoresis studies of native protein were performed in vertical slabs of polyacrylamide gel (PAGE), in an alkaline discontinuous buffer system, with a concentration of 10% in the separation gel. SDS-PAGE was performed in PhastGel® (8-25). Both gels were stained with Coomassie Blue. Gels were analyzed using the VDS-Pharmacia® device. Our results indicate that all analyzed venom samples showed different protein composition, although common protein fractions were detected in some individual samples. Differences were observed between the different individual venom samples and so in the same specimen in relation to the time of collection, for both techniques used. Diet did not influence the variability of venom composition. There is a significant difference in native venom protein composition of males and females.

Crotalus durissus terrificus; intraspecific variation; protein composition; venom; sex; feeding; electrophoresis

Short communication

INTRASPECIFIC VARIATION IN THE VENOM ELECTROPHORETIC PROFILE OF RECENTLY CAPTURED Crotalus durissus terrificus (LAURENTI, 1768) SNAKES

A. J. MAGRO¹, R. J. DA SILVA¹, P. R. R. RAMOS², A. L. CHERUBINI¹, M. R. HATAYDE³

1 Centro de Estudos de Venenos e Animais Peçonhentos da UNESP (CEVAP/UNESP), Distrito de Rubião Júnior, Botucatu, São Paulo, Brasil; 2 Departamento de Física e Biofísica, Instituto de Biociências, UNESP, Botucatu, São Paulo, Brasil, 3 Departamento de Clínica Veterinária, Faculdade de Ciências Agrárias e Veterinárias, UNESP, Jaboticabal, São Paulo, Brasil.

References

01 AMARAL A. Serpentes do Brasil: iconografia colorida. São Paulo: Melhoramentos, Edusp, 1978: 247p.
02 BARRIO A. Estudio electroforético del veneno de serpiente de cascabel. Cienc. Invest., 1954, 10, 368-70.
03 BARRIO A., BRAZIL O. Neuromuscular action of the Crotalus terrificus terrificus venoms. Acta Physiol. Latinoam., 1951, 1, 291-308.
04 BERCOVICI D., CHUDZINSKI AM., DIAS WO., ESTEVES MI., HIRAICHI E., OISHI NY., PICARELLI ZP., ROCHA MC., UEDA CMPM., YAMANOUYE N., RAW I. A systematic fractionation of Crotalus durissus terrificus venom.Mem. Inst. Butantan (São Paulo), 1987, 49, 69-78.
05 CAMPBELL JA., LAMAR WW. The venomous reptiles of Latin America. New York: Cornell University Press, 1989: 425p.
06 CHIPPAUX J., WILLIAMS V., WHITE J. Snake venom variability: methods of study results and interpretation. Toxicon, 1991, 29, 1279-303.
07 DALTRY JC., WÜSTER W., THORPE RS. Diet and snake evolution. Nature, 1996, 379, 537-40.
08 FIERO MK., SEIFERT MW., WEAVER JJ., BONILLA CA. Comparative study of juvenile and adult prairie rattlesnake (Crotalus viridis viridis) venoms.Toxicon, 1972, 10, 81-2.
09 FRANCISCHETTI IM., GOMBAROVITS ME., VALENZUELA JG., CARLINI CR., GUIMARÃES JA. Intraspecific variation in the venoms of the South American rattlesnake (Crotalus durissus terrificus). Comp. Biochem. Physiol. C, 2000, 127, 23-36.
¹⁰ GAHNE B., JUNEJA RK., GROUMUS J. Horizontal polyacrylamide gradient gel electrophoresis for the simultaneous phenotyping of transferrin post-transferrin albumin and post-albumin in the blood of cattle. Anim. Blood Groups Biochem. Genet., 1977, 8, 127-37.
¹¹ GIORGI R., BERNARDI MM., CURY Y. Analgesic effect evoked by low molecular weight substances extracted from Crotalus durissus terrificus venom. Toxicon, 1993, 31: 1257-65.
¹² GLENN JL., STRAIGHT R. Mojave rattlesnake Crotalus scutulatus scutulatus venom: variation in toxicity with geographical origin. Toxicon, 1978, 16, 81-4.
¹³ GONÇALVES JM., VIEIRA LG. Estudos sobre venenos de serpentes brasileiras. I. Análise eletroforética. An. Acad. Bras. Ciênc., 1950, 22, 141-50.
¹⁴ GUBENSEK F., SKET D., TURK V., LEBEZ D. Fractionation of Vipera ammodytes venom and seasonal variation of its composition. Toxicon, 1974, 12, 167-71.
¹⁵ HAMES BD., RICKWOOD D. Gel electrophoresis of proteins 2.ed. New York: Oxford University, 1990: 383p.
¹⁶ IRWIN RL., OLIVIER KL., MOHAMED AH., HAAST WE. Toxicity of elapid venoms and observation in relation to geographical location. Toxicon, 1970, 8, 51-4.
¹⁷ JIMENEZ-PORRAS JM. Intraspecific variations in composition of venom of the jumping viper Bothrops nummifera Toxicon, 1964, 2, 187-96.
¹⁸ LATIFI M. Variation in yield and lethality of venom from Iranian snakes. Toxicon, 1984, 22, 373-80.
¹⁹ MANCIN AC., SOARES AM., ANDRIÃO-ESCARSO SH., FAÇA V., GREENE LG., ZUCOLLOTTO S., PELÁ I., GIGLIO JR. The analgesic activity of crotamine, a neurotoxin from Crotalus durissus terrificus (South American rattlesnake) venom: a biochemical and pharmacological study.Toxicon, 1998, 36, 1927-37.
²⁰ MARSH A., GLATSTON A. Some observations on the venom of the Rhinoceros horned viper Bitis nasicornis (Shaw). Toxicon, 1974, 12, 621-8.
²¹ MATTISON C. Rattler! - A natural history of rattlesnakes. London: Blandford, 1996: 144p.
²² MEBS D., KORNALIK F. Intraspecific variation in content of a basic toxin in eastern diamondback rattlesnakes (Crotalus adamanteus) venom. Toxicon, 1984, 22, 831-3.
²³ RODRIGUES VM., SOARES AM., MANCIN AC., FONTES MRM., HOMSI-BRANDENBURGO MI., GIGLIO JR. Geographic variations in the composition of myotoxins from Bothrops neuwiedi snake venoms: biochemical characterization and biological activity. Comp. Biochem. Physiol. A, 1998, 121, 215-22.
²⁴ SCHENBERG S. Geographical pattern of crotamine distribution in the same rattlesnake subspecies. Science, 1959, 129, 1361-3.
²⁵ SILVA R. J. Efeito dos venenos de serpentes Crotalus durissus terrificus (LAURENTI 1768) e Bothrops jararaca (WIED 1824) na evolução do tumor ascítico de Ehrlich. Botucatu: Universidade Estadual Paulista, Faculdade de Medicina, 1995. 132p. [Dissertação - Mestrado].
²⁶ SILVA RJ., FECCHIO D., BARRAVIERA B. Antitumor effect of snake venoms.J. Venom. Anim. Toxins, 1996, 2, 79-90.
²⁷ SILVA RJ., FECCHIO D., BARRAVIERA B. Effect of Crotalus durissus terrificus venom on evolution of the Ehrlich ascites tumor. J. Venom. Anim. Toxins, 1997, 3, 324-41.
²⁸ SOARES AM., ANZALONI PEDROSA LH., FONTES MRM., SILVA RJ., GIGLIO JR. Polyacrylamide gel electrophoresis as a tool for the taxonomic identification of snakes from the Elapidae and Viperidae families. J. Venom. Anim. Toxins, 1998, 4, 137-42.
²⁹ SOERENSEN B. Animais peçonhentos: reconhecimento distribuição geográfica produção de soros clínica tratamento dos envenenamentos. São Paulo: Atheneu, 1990. 138p.
³⁰ SOLORZANO A., ROMERO M., GUTIÉRREZ JM., SASA M. Venom composition and diet of the cantil Agkistrodon bilineatus howardgloydi (Serpentes: Viperidae). Southwest. Nat., 1999, 44, 478-83.
³¹ STOLF HO., BARRAVIERA SRCS., THOMAZINI I. Cola de fibrina derivada de veneno de cobra: uso experimental em cirurgia dermatológica. Nota prévia. In: CONGRESSO BRASILEIRO DE DERMATOLOGIA, 48, Curitiba, 1997. Abstracts... Curitiba: 1997. 168.
³² TABORSKA E. Intraspecies variability of the venom of Echis carinatus. Physiol. Bohemoslov., 1971, 20, 307-18.
³³ VILLELA GG., BACILA M., TASTALDI H. Técnicas e experimentos de Bioquímica. Rio de Janeiro: Guanabara Koogan, 1972. 552p
³⁴ WILLEMSE GT. Individual variation in snake venom. Comp. Biochem. Physiol. B, 1978, 61, 553-7.

Publication Dates

Publication in this collection
08 Oct 2002
Date of issue
Dec 2001

History

Received
18 Jan 2001
Accepted
09 Mar 2000

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

[1] 01 AMARAL A. Serpentes do Brasil: iconografia colorida. São Paulo: Melhoramentos, Edusp, 1978: 247p.

[2] 02 BARRIO A. Estudio electroforético del veneno de serpiente de cascabel. Cienc. Invest., 1954, 10, 368-70.

[3] 03 BARRIO A., BRAZIL O. Neuromuscular action of the Crotalus terrificus terrificus venoms. Acta Physiol. Latinoam., 1951, 1, 291-308.

[4] 04 BERCOVICI D., CHUDZINSKI AM., DIAS WO., ESTEVES MI., HIRAICHI E., OISHI NY., PICARELLI ZP., ROCHA MC., UEDA CMPM., YAMANOUYE N., RAW I. A systematic fractionation of Crotalus durissus terrificus venom.Mem. Inst. Butantan (São Paulo), 1987, 49, 69-78.

[5] 05 CAMPBELL JA., LAMAR WW. The venomous reptiles of Latin America. New York: Cornell University Press, 1989: 425p.

[6] 06 CHIPPAUX J., WILLIAMS V., WHITE J. Snake venom variability: methods of study results and interpretation. Toxicon, 1991, 29, 1279-303.

[7] 07 DALTRY JC., WÜSTER W., THORPE RS. Diet and snake evolution. Nature, 1996, 379, 537-40.

[8] 08 FIERO MK., SEIFERT MW., WEAVER JJ., BONILLA CA. Comparative study of juvenile and adult prairie rattlesnake (Crotalus viridis viridis) venoms.Toxicon, 1972, 10, 81-2.

[9] 09 FRANCISCHETTI IM., GOMBAROVITS ME., VALENZUELA JG., CARLINI CR., GUIMARÃES JA. Intraspecific variation in the venoms of the South American rattlesnake (Crotalus durissus terrificus). Comp. Biochem. Physiol. C, 2000, 127, 23-36.

[10] ¹⁰ GAHNE B., JUNEJA RK., GROUMUS J. Horizontal polyacrylamide gradient gel electrophoresis for the simultaneous phenotyping of transferrin post-transferrin albumin and post-albumin in the blood of cattle. Anim. Blood Groups Biochem. Genet., 1977, 8, 127-37.

[11] ¹¹ GIORGI R., BERNARDI MM., CURY Y. Analgesic effect evoked by low molecular weight substances extracted from Crotalus durissus terrificus venom. Toxicon, 1993, 31: 1257-65.

[12] ¹² GLENN JL., STRAIGHT R. Mojave rattlesnake Crotalus scutulatus scutulatus venom: variation in toxicity with geographical origin. Toxicon, 1978, 16, 81-4.

[13] ¹³ GONÇALVES JM., VIEIRA LG. Estudos sobre venenos de serpentes brasileiras. I. Análise eletroforética. An. Acad. Bras. Ciênc., 1950, 22, 141-50.

[14] ¹⁴ GUBENSEK F., SKET D., TURK V., LEBEZ D. Fractionation of Vipera ammodytes venom and seasonal variation of its composition. Toxicon, 1974, 12, 167-71.

[15] ¹⁵ HAMES BD., RICKWOOD D. Gel electrophoresis of proteins 2.ed. New York: Oxford University, 1990: 383p.

[16] ¹⁶ IRWIN RL., OLIVIER KL., MOHAMED AH., HAAST WE. Toxicity of elapid venoms and observation in relation to geographical location. Toxicon, 1970, 8, 51-4.

[17] ¹⁷ JIMENEZ-PORRAS JM. Intraspecific variations in composition of venom of the jumping viper Bothrops nummifera Toxicon, 1964, 2, 187-96.

[18] ¹⁸ LATIFI M. Variation in yield and lethality of venom from Iranian snakes. Toxicon, 1984, 22, 373-80.

[19] ¹⁹ MANCIN AC., SOARES AM., ANDRIÃO-ESCARSO SH., FAÇA V., GREENE LG., ZUCOLLOTTO S., PELÁ I., GIGLIO JR. The analgesic activity of crotamine, a neurotoxin from Crotalus durissus terrificus (South American rattlesnake) venom: a biochemical and pharmacological study.Toxicon, 1998, 36, 1927-37.

[20] ²⁰ MARSH A., GLATSTON A. Some observations on the venom of the Rhinoceros horned viper Bitis nasicornis (Shaw). Toxicon, 1974, 12, 621-8.

[21] ²¹ MATTISON C. Rattler! - A natural history of rattlesnakes. London: Blandford, 1996: 144p.

[22] ²² MEBS D., KORNALIK F. Intraspecific variation in content of a basic toxin in eastern diamondback rattlesnakes (Crotalus adamanteus) venom. Toxicon, 1984, 22, 831-3.

[23] ²³ RODRIGUES VM., SOARES AM., MANCIN AC., FONTES MRM., HOMSI-BRANDENBURGO MI., GIGLIO JR. Geographic variations in the composition of myotoxins from Bothrops neuwiedi snake venoms: biochemical characterization and biological activity. Comp. Biochem. Physiol. A, 1998, 121, 215-22.

[24] ²⁴ SCHENBERG S. Geographical pattern of crotamine distribution in the same rattlesnake subspecies. Science, 1959, 129, 1361-3.

[25] ²⁵ SILVA R. J. Efeito dos venenos de serpentes Crotalus durissus terrificus (LAURENTI 1768) e Bothrops jararaca (WIED 1824) na evolução do tumor ascítico de Ehrlich. Botucatu: Universidade Estadual Paulista, Faculdade de Medicina, 1995. 132p. [Dissertação - Mestrado].

[26] ²⁶ SILVA RJ., FECCHIO D., BARRAVIERA B. Antitumor effect of snake venoms.J. Venom. Anim. Toxins, 1996, 2, 79-90.

[27] ²⁷ SILVA RJ., FECCHIO D., BARRAVIERA B. Effect of Crotalus durissus terrificus venom on evolution of the Ehrlich ascites tumor. J. Venom. Anim. Toxins, 1997, 3, 324-41.

[28] ²⁸ SOARES AM., ANZALONI PEDROSA LH., FONTES MRM., SILVA RJ., GIGLIO JR. Polyacrylamide gel electrophoresis as a tool for the taxonomic identification of snakes from the Elapidae and Viperidae families. J. Venom. Anim. Toxins, 1998, 4, 137-42.

[29] ²⁹ SOERENSEN B. Animais peçonhentos: reconhecimento distribuição geográfica produção de soros clínica tratamento dos envenenamentos. São Paulo: Atheneu, 1990. 138p.

[30] ³⁰ SOLORZANO A., ROMERO M., GUTIÉRREZ JM., SASA M. Venom composition and diet of the cantil Agkistrodon bilineatus howardgloydi (Serpentes: Viperidae). Southwest. Nat., 1999, 44, 478-83.

[31] ³¹ STOLF HO., BARRAVIERA SRCS., THOMAZINI I. Cola de fibrina derivada de veneno de cobra: uso experimental em cirurgia dermatológica. Nota prévia. In: CONGRESSO BRASILEIRO DE DERMATOLOGIA, 48, Curitiba, 1997. Abstracts... Curitiba: 1997. 168.

[32] ³² TABORSKA E. Intraspecies variability of the venom of Echis carinatus. Physiol. Bohemoslov., 1971, 20, 307-18.

[33] ³³ VILLELA GG., BACILA M., TASTALDI H. Técnicas e experimentos de Bioquímica. Rio de Janeiro: Guanabara Koogan, 1972. 552p

[34] ³⁴ WILLEMSE GT. Individual variation in snake venom. Comp. Biochem. Physiol. B, 1978, 61, 553-7.