Screening the mutagenic activities of coomonly used antiparasite drugs by the Simultest, a simplified Salmonella/microsome plate incorporation assay

Melo, Maria Eliane B.; Ferreira, Luís Carlos S.

doi:10.1590/S0036-46651990000400006

Abstracts

The mutagenic activities of 16 anti-parasite drugs were screened by the Simultest in both qualitative (spot test) and quantitative (plate incorporation) assays with a Salmonella typhimurium pool composed by the indicator strains TA97, TA 98, TA100 and TA102. Four anti Chagas' disease drugs (nifurtimox, benznidazole, CL 64,855, and MK 436) and two anti-amebae drugs (metronidazole and tinidazole) gave positive results in qualitative tests and incorporation of rat liver microsomes did not alter the results. Comparative dose response curves of the mutagenic activities of CL 64,855, metronidazole and benznidazole obtained by the simultest and by individual Salmonella indicator strains demonstrated that both approachs have similar sensitivities. The results corroborate the validity of the Simultest, as a simplified, fast and economic version of the Ames test in preliminary screening of potential mutagenic drugs.

Simultest; Anti-parasite drugs; Ames test

As atividades mutagênicas de 16 drogas com ação anti-parasitária foram avaliadas pelo Simultest em ensaios qualitativos (spot testes) e quantitativos (incorporação em placa) com uma mistura das linhagens indicadoras de Salmonella typhimurium TA97, TA98, TA100 e TA102. Quatro drogas anti-doença de Chagas (nifurtimox, benzonidazol, CL 64,855 e MK 436) e duas drogas anti-amebiase (metronidazol e tinidazol) deram resultados positivos em testes qualitativos e a incorporação de fração microssomal de fígado de rato não alterou os resultados. Curvas comparadas de efeito da dose da atividade mutagênica do metronidazol, benzonidazol e CL 64,855 detectadas pelo Simultest e linhagens indicadoras individuais demonstraram que as duas abordagens possuem sensibilidades semelhantes. Os resultados corroboram a validade do Simultest como uma versão simplificada, rápida e econômica do teste de Ames no rastreamento preliminar de drogas potencialmente mutagênicas.

ORIGINAL ARTICLES

Screening the mutagenic activities of coomonly used antiparasite drugs by the Simultest, a simplified Salmonella/microsome plate incorporation assay

Rastreamento de atividade mutagênica de drogas anti-parasitárias de uso comum pelo Simultest, uma versão simplificada do teste Salmonella/fração microssomal

Maria Eliane B. Melo^I; Luís Carlos S. Ferreira^II,III

^ICentro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz. Recife, Pernambuco, Brazil

^IIDepartamento de Biofísica e Radiobiologia and Laboratório de Imunopatologia Keizo Asami, Universidade Federal de Pernambuco. Recife, Pernambuco, Brazil

^IIIPresent address: Instituto de Biofísica Carlos Chagas Filho. Universidade Federal do Rio de Janeiro CCS. Cidade Universitária, Ilha do Fundão. 21949 Rio de Janeiro. RJ, Brazil

^{All correspondence should be sent to} All correspondence should be sent to: Luís Carlos S. Ferreira

SUMMARY

The mutagenic activities of 16 anti-parasite drugs were screened by the Simultest in both qualitative (spot test) and quantitative (plate incorporation) assays with a Salmonella typhimurium pool composed by the indicator strains TA97, TA 98, TA100 and TA102. Four anti Chagas' disease drugs (nifurtimox, benznidazole, CL 64,855, and MK 436) and two anti-amebae drugs (metronidazole and tinidazole) gave positive results in qualitative tests and incorporation of rat liver microsomes did not alter the results. Comparative dose response curves of the mutagenic activities of CL 64,855, metronidazole and benznidazole obtained by the simultest and by individual Salmonella indicator strains demonstrated that both approachs have similar sensitivities. The results corroborate the validity of the Simultest, as a simplified, fast and economic version of the Ames test in preliminary screening of potential mutagenic drugs.

Key words: Simultest; Anti-parasite drugs; Ames test.

RESUMO

As atividades mutagênicas de 16 drogas com ação anti-parasitária foram avaliadas pelo Simultest em ensaios qualitativos (spot testes) e quantitativos (incorporação em placa) com uma mistura das linhagens indicadoras de Salmonella typhimurium TA97, TA98, TA100 e TA102. Quatro drogas anti-doença de Chagas (nifurtimox, benzonidazol, CL 64,855 e MK 436) e duas drogas anti-amebiase (metronidazol e tinidazol) deram resultados positivos em testes qualitativos e a incorporação de fração microssomal de fígado de rato não alterou os resultados. Curvas comparadas de efeito da dose da atividade mutagênica do metronidazol, benzonidazol e CL 64,855 detectadas pelo Simultest e linhagens indicadoras individuais demonstraram que as duas abordagens possuem sensibilidades semelhantes. Os resultados corroboram a validade do Simultest como uma versão simplificada, rápida e econômica do teste de Ames no rastreamento preliminar de drogas potencialmente mutagênicas.

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

ACKNOWLEDGMENTS

We acknowledge the excelent technical assistence of Mr. S. H. Almeida and Dr. Ames and Dr. Rosenkranz for the bacterial indicator strains.

Recebido para publicação em 23/10/1989

Research supported by the Japanese International Cooperation Agency, FINEP and CNPq.

1. AMES, B.N. Detection of environmental mutagens and potential carcinogens. Cancer, 53: 2034-2040, 1984.
2. AMES, B.N. Mutagenesis and carcinogenesis: endogenous and exogenous factors. Environ. Mol. Mutagen., 14: 66-77, 1989.
3. AMES, B.N.; McCANN, J. & YAMASAHI, E. Methods for detecting carcinogens and mutagens with the Salmonella/mammalian microsome mutagenicity test. Mutation Res., 31: 347-363, 1975.
4. ANDRADE, S.G.; SILVA, R.C.; SANTIAGO, C.M.G. & FREITAS, L.A.R. Therapeutic action of MK-436 (2,5-nitroimidazole) on Trypanosoma cruzi infection in mice a parasitological, serological, histopathological and structural study. Bull. Wld. Hlth. Org., 65: 625-633, 1987.
5. ARRIAGA ALBA, M.; ESPINOSA, J.E. & CORTINAS DE NAVA, C. Mutagenicity of products generated by the reaction between several antiparasitic drugs and nitrite. Environ. Mol. Mutagen., 12: 65-73, 1988.
6. EBRINGER, L. & BENCOVA, M. Mutagenicity of nitrofuran drugs in bacterial systems. Folia microbiol. (Praha), 25: 388-396, 1980.
7. ESPINOSA-AGUIRRE, J.J.; C. AROUMIR, C.; MEZA, M.T.; CIENFUEGOS, E. & CORTINAS DE NAVA, C. Genotoxicity of amebicide anthelmintic drugs in Escherichia coli polA+/polA-. Mutation Res., 188: 111-120, 1987.
8. FERREIRA, R.C.C. & FERREIRA, L.C.S. Mutagenicity of mifurtimox and benznidazole in the Salmonella/microsome assay. Braz. J. med. biol. Res., 19: 19-25, 1986.
9. FILARDI, L.S. & BRENER, Z. A nitroimidazole-thiadizale derivative with curative action in experimental Trypanosoma cruzi infections. Ann. trop. Med. Parasit., 76: 293-297, 1982.
10. HARTMAN, P.E.; HULBERT, P.B.; BUEDING, E. & TAYLOR, D.D. Microsomal activation to mutagens of antischistosomal methyl thioxanthenones and initial tests on a possibly non-mutagenic analogue. Mutation Res., 31: 87-95, 1975.
11. MARON, D.M. & AMES, B.N. Revised methods for the Salmonella mutagenicity test. Mutation Res., 113: 173-215, 1983.
12. McCANN, J.; SPINGARN, N.E.; KOBORI, J. & AMES, B.N. Detection of carcinogens as mutagens: bacterial tests strains with R factors plasmids. Proc. nat. Acad. Sci. (Wash.), 72: 979-983, 1975a.
13. McCANN, J.; CHOI, E.; YAMASAKI, E. & AMES, B.N. Detection of carcinogens as mutagens in the Salmonella microsome test: assay of 300 chemicals. Proc. nat. Acad. Sci. (Wash.), 72: 5135-5139, 1975b.
14. NESTMANN, E.R.; BRILLINGER, R.L.; McPHERSON, M.F. & MARS, K.L. The Simultest: a new approach to screening chemicals with the Salmonella reversion assay. Environ. Mol. Mutagen., 10: 169-181, 1987.
15. NETO, V.A. & BALDY, J.L. Doenças transmissíveis. Săo Paulo, Sarvier, 1989.
16. SPECK, W.T.; STEIN, A.B. & ROSENKRANZ, H.S. Mutagenicity of metronidazole: presence of several active metabolites in human urine. J. nat. Cancer Inst., 56: 283-284, 1976.
17. ZIEGER, E.; RISKO, K.J. & MARGOLIN, B.H. Strategies to reduce the cost of mutagenicity screening with the Salmonella assay. Environ. Mol. Mutagen., 7: 901-911, 1985.

All correspondence should be sent to:

Luís Carlos S. Ferreira

Publication Dates

Publication in this collection
12 Sept 2006
Date of issue
Aug 1990

History

Received
23 Oct 1989

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

[1] 1. AMES, B.N. Detection of environmental mutagens and potential carcinogens. Cancer, 53: 2034-2040, 1984.

[2] 2. AMES, B.N. Mutagenesis and carcinogenesis: endogenous and exogenous factors. Environ. Mol. Mutagen., 14: 66-77, 1989.

[3] 3. AMES, B.N.; McCANN, J. & YAMASAHI, E. Methods for detecting carcinogens and mutagens with the Salmonella/mammalian microsome mutagenicity test. Mutation Res., 31: 347-363, 1975.

[4] 4. ANDRADE, S.G.; SILVA, R.C.; SANTIAGO, C.M.G. & FREITAS, L.A.R. Therapeutic action of MK-436 (2,5-nitroimidazole) on Trypanosoma cruzi infection in mice a parasitological, serological, histopathological and structural study. Bull. Wld. Hlth. Org., 65: 625-633, 1987.

[5] 5. ARRIAGA ALBA, M.; ESPINOSA, J.E. & CORTINAS DE NAVA, C. Mutagenicity of products generated by the reaction between several antiparasitic drugs and nitrite. Environ. Mol. Mutagen., 12: 65-73, 1988.

[6] 6. EBRINGER, L. & BENCOVA, M. Mutagenicity of nitrofuran drugs in bacterial systems. Folia microbiol. (Praha), 25: 388-396, 1980.

[7] 7. ESPINOSA-AGUIRRE, J.J.; C. AROUMIR, C.; MEZA, M.T.; CIENFUEGOS, E. & CORTINAS DE NAVA, C. Genotoxicity of amebicide anthelmintic drugs in Escherichia coli polA+/polA-. Mutation Res., 188: 111-120, 1987.

[8] 8. FERREIRA, R.C.C. & FERREIRA, L.C.S. Mutagenicity of mifurtimox and benznidazole in the Salmonella/microsome assay. Braz. J. med. biol. Res., 19: 19-25, 1986.

[9] 9. FILARDI, L.S. & BRENER, Z. A nitroimidazole-thiadizale derivative with curative action in experimental Trypanosoma cruzi infections. Ann. trop. Med. Parasit., 76: 293-297, 1982.

[10] 10. HARTMAN, P.E.; HULBERT, P.B.; BUEDING, E. & TAYLOR, D.D. Microsomal activation to mutagens of antischistosomal methyl thioxanthenones and initial tests on a possibly non-mutagenic analogue. Mutation Res., 31: 87-95, 1975.

[11] 11. MARON, D.M. & AMES, B.N. Revised methods for the Salmonella mutagenicity test. Mutation Res., 113: 173-215, 1983.

[12] 12. McCANN, J.; SPINGARN, N.E.; KOBORI, J. & AMES, B.N. Detection of carcinogens as mutagens: bacterial tests strains with R factors plasmids. Proc. nat. Acad. Sci. (Wash.), 72: 979-983, 1975a.

[13] 13. McCANN, J.; CHOI, E.; YAMASAKI, E. & AMES, B.N. Detection of carcinogens as mutagens in the Salmonella microsome test: assay of 300 chemicals. Proc. nat. Acad. Sci. (Wash.), 72: 5135-5139, 1975b.

[14] 14. NESTMANN, E.R.; BRILLINGER, R.L.; McPHERSON, M.F. & MARS, K.L. The Simultest: a new approach to screening chemicals with the Salmonella reversion assay. Environ. Mol. Mutagen., 10: 169-181, 1987.

[15] 15. NETO, V.A. & BALDY, J.L. Doenças transmissíveis. Săo Paulo, Sarvier, 1989.

[16] 16. SPECK, W.T.; STEIN, A.B. & ROSENKRANZ, H.S. Mutagenicity of metronidazole: presence of several active metabolites in human urine. J. nat. Cancer Inst., 56: 283-284, 1976.

[17] 17. ZIEGER, E.; RISKO, K.J. & MARGOLIN, B.H. Strategies to reduce the cost of mutagenicity screening with the Salmonella assay. Environ. Mol. Mutagen., 7: 901-911, 1985.

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