Biochemical behavior of Trypanosoma cruzi strains isolated from mice submitted to specific chemotherapy

Marretto, Jesila Pinto M.; Andrade, Sonia G.

doi:10.1590/S0037-86821994000400002

Abstracts

To investigate the influence of chemotherapy on the biochemical beha vior of Trypanosoma cruzi strains, three groups of mice were infected with one of three strains of T. cruzi of different biological and isoenzymic patterns (Peruvian, 21 SF and Colombian strains). Each group was subdivided into subgroups: 1 - treated with nifurtimox; 2 - treated with benznidazole and 3 - untreated infected controls. At the end of treatment, that lasted for 90 days, xenodiagnosis, sub inoculation of blood into new born mice and haemoculture were performed as tests of cure. From the positive tests, 22 samples of T. cruzi were isolated from all subgroups. Electrophoretic analysis of the isoenzymes PGM, GP1, ALAT and AS AT failed to show any difference between parasite strains isolated from treated and untreated mice, which indicates that no detectable clonal selection or parasite genetic markers alterations concerning the isoenzymes analysed have been determined by treatment with drugs of recognized antiparasitic effect, suggesting stability of the phenotypic characteristics of the three biological types of T. cruzi strains.

Trypanosoma cruzi strains; Isoenzymes; Chemotherapy

Com o objetivo de investigar a influência da quimioterapia no padrão bioquímico de diferentes cepas do Trypanosoma cruzi, três grupos de camundongos foram infectados respectivamente com as cepas Peruana, 21 SF e Colombiana, que correspondem a diferentes padrões biológicos e isoenzimáticos. Cada grupo foi subdividido em subgrupos: 1 - tratados com nifurtimox; 2 - tratados com benzonidazol; 3- controles infectados não tratados. Ao final do tratamento que durou 90 dias, os animais foram submetidos a testes parasitológicos de cura: xenodiagnóstico, subinoculação do sangue em camundongos recém-nascidos e hemocultura em meio Warren. A partir da positivação destes testes, foram isoladas 22 amostras do T. cruzi dos três subgrupos. A análise eletroforética dos extratos enzimáticos obtidos após cultura para as enzimas PGM, GPI, ALAT e ASAT demonstrou identidade dos padrães enzimáticos entre os parasitos de uma mesma cepa isolados de animais tratados ou não tratados. Conclui-se que não houve seleção clonal ou alterações genéticas dos parasitos, detectáveis pelas isoenzimas examinadas, dependentes da ação de drogas de reconhecida ação antiparasitária. Isto sugere uma estabilidade dos caracteres fenotípicos dos três tipos biológicos de cepas do T. cruzi.

Cepas do Trypanosoma cruzi; Isoenzimas; Quimioterapia

ARTICLES

Biochemical behavior of Trypanosoma cruzi strains isolated from mice submitted to specific chemotherapy

Jesila Pinto M. Marretto; Sonia G. Andrade

^{Adress to correspondence} Adress to correspondence: Dr a Sonia G. Andrade. Centro de Pesquisas Gonçalo Moniz/FIOCRUZ/UFBA. R. Valdemar Falcão 121 Brotas, 40295-001 Salvador, BA, Brasil.

ABSTRACT

To investigate the influence of chemotherapy on the biochemical beha vior of Trypanosoma cruzi strains, three groups of mice were infected with one of three strains of T. cruzi of different biological and isoenzymic patterns (Peruvian, 21 SF and Colombian strains). Each group was subdivided into subgroups: 1 - treated with nifurtimox; 2 - treated with benznidazole and 3 - untreated infected controls. At the end of treatment, that lasted for 90 days, xenodiagnosis, sub inoculation of blood into new born mice and haemoculture were performed as tests of cure. From the positive tests, 22 samples of T. cruzi were isolated from all subgroups. Electrophoretic analysis of the isoenzymes PGM, GP1, ALAT and AS AT failed to show any difference between parasite strains isolated from treated and untreated mice, which indicates that no detectable clonal selection or parasite genetic markers alterations concerning the isoenzymes analysed have been determined by treatment with drugs of recognized antiparasitic effect, suggesting stability of the phenotypic characteristics of the three biological types of T. cruzi strains.

Keywords:Trypanosoma cruzi strains. Isoenzymes. Chemotherapy.

RESUMO

Com o objetivo de investigar a influência da quimioterapia no padrão bioquímico de diferentes cepas do Trypanosoma cruzi, três grupos de camundongos foram infectados respectivamente com as cepas Peruana, 21 SF e Colombiana, que correspondem a diferentes padrões biológicos e isoenzimáticos. Cada grupo foi subdividido em subgrupos: 1 - tratados com nifurtimox; 2 - tratados com benzonidazol; 3- controles infectados não tratados. Ao final do tratamento que durou 90 dias, os animais foram submetidos a testes parasitológicos de cura: xenodiagnóstico, subinoculação do sangue em camundongos recém-nascidos e hemocultura em meio Warren. A partir da positivação destes testes, foram isoladas 22 amostras do T. cruzi dos três subgrupos. A análise eletroforética dos extratos enzimáticos obtidos após cultura para as enzimas PGM, GPI, ALAT e ASAT demonstrou identidade dos padrães enzimáticos entre os parasitos de uma mesma cepa isolados de animais tratados ou não tratados. Conclui-se que não houve seleção clonal ou alterações genéticas dos parasitos, detectáveis pelas isoenzimas examinadas, dependentes da ação de drogas de reconhecida ação antiparasitária. Isto sugere uma estabilidade dos caracteres fenotípicos dos três tipos biológicos de cepas do T. cruzi.

Palavras-chave: Cepas do Trypanosoma cruzi. Isoenzimas. Quimioterapia.

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Recebido para publicação em 25/03/94.

Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Universidade Federal da Bahia, Salvador, BA.

1. Andrade SG. Caracterização de cepas do Trypanosoma cruzi isoladas do Recôncavo Bahiano. Revista de Patologia Tropical 3:65-121, 1974.
2. Andrade SG. Morphological and behavioral characterization of Trypanosoma cruzi strains. Revista da Sociedade Brasileira de Medicina Tropical 18(suppl):39-46, 1985.
3. Andrade SG, Andrade, ZA, Figueira RM. Estudo experimental sobre a resistência de uma cepa do Trypanosoma cruzi ao Bay 2502. Revistado Instituto de Medicina Tropical de São Paulo 19:124-129,1977.
4. Andrade SG, Figueira RM. Estudo experimental sobre a ação terapêutica da droga R07-1051 na infeção por diferentes cepas do Trypanosoma cruzi Revista do Instituto de Medicina Tropical de São Paulo 9:335-341, 1977.
5. Andrade SG, Figueira RM, Carvalho ML, Gorini DF. Influência da cepa do Trypanosoma cruzi na resposta à terapêutica pelo Bay 2502 (Resultados de tratamento a longo prazo). Revista do Instituto de Medicina Tropical de São Paulo 17:380-389, 1975.
6. Andrade SG, Magalhães JB, Pontes AL. Evaluation of chemotherapy with benznidazole and nifurtimox in mice infected with Trypanosoma cruzi strains of different types. Bulletin of the World Health Organization 63:721-726, 1985.
7. Andrade V, Brodskyn C, Andrade SG. Correlation between isoenzymepatterns andbiologicalbehavior of different strains of Trypanosoma cruzi Transactions of the Royal Society of Tropical Medicine and Hygiene 77:796-799, 1983.
8. Bogliolo AR, Chiari E, Silva-Pereira RO, Silva Pereira AA. A comparative study of Trypanosoma cruzi enzyme polymorphism in South America. Brazilian JoumalofMedical and Bio logical Research 19: 673-683, 1986.
9. Bogliolo AR, Lanham SM, Corredor A. Further enzyme polymorphism in Trypanosoma cruzi from Colombia. Brazilian Journal of Medical and Biological Research 18: 427-433, 1985.
10. Brener Z, Costa CAG, Chiari C. Differences in the susceptibility of Trypanosoma cruzi strains to active chemotherapeutic agents. Revista do Instituto de Medicina Tropical de São Paulo 18:450-455, 1976.
11. Bongertz V, Dvorak JA. Trypanosoma cruzi: antigenic analysis of cloned stocks. The American Journal of Tropical Medicine and Hygiene 32:716- 722, 1983.
12. Carneiro M, Chiari E, Gonçalves AM, Silva Pereira AA, Morel CM, Romanha AJ. Changes in the isoenzyme and kinetoplast DNA patterns of Trypanosoma cruzi strains induced by maintenance in mice. Acta Tropica 47:35-45, 1990.
13. Castro Silva R, Santiago CMG, AndradeSG. Estudo da estabilidade dos caracteres isoenzimáticos de cepas do Trypanosoma cruzi em extratos enzimáticos estocados em nitrogênio líquido, por período prolongado. Memórias do Instituto Oswaldo Cruz 83(supl I):82, 1988.
14. Castro Silva R, Santiago CMG, Pontes AL, Andrade SG. Padrão isoenzimático da cepa Y do Trypanosoma cruzi após quimioterapia específica. Memórias do Instituto Oswaldo Cruz 84: 81-86, 1989.
15. Chapman MD, Baggaley RC, Godfrey-Fausset P, Malpas TJ, White G, Canese J, Miles MAS. Trypanosoma cruzi from the Paraguayan chaco: isoenzyme profiles of strains isolated at Makthlawaiya. Journal of Protozoology 31:482- 486, 1984.
16. Dvorak JA, Hartman DL, Miles MA. Trypanosoma cruzi: Correlation of growth kinetics to zymodeme type in clones derived from various sources. Joumal of Protozoology 27:472-474, 1980.
17. FedericiEE, AbelmannWB,NevaFA. Chronicand progressive myocarditis in C3H mice infected with Trypanosoma cruzi The American Journal of Tropical Medicine and Hygiene 13:272-280, 1964.
18. Godfrey DG, Kilgour V. Enzyme electrophoresis in characterizing the causative organism of gambian trypanosomiasis. Transactions of the Royal Society ofTropicalMedicineand Hygiene 3:219-224,1976.
19. Goldberg SS , Silva Pereira AA. Enzyme variation among clones of Trypanosoma cruzi Journal of Parasitology 69:91-96, 1983.
20. Magalhães JB, Andrade SG. Estudo do comportamento de cepas de Trypanosoma cruzi após passagem em diferentes espécies de triatomíneos. Revista da Sociedade Brasileira de Medicina Tropical 24:209-216, 1991.
21. Magalhães JB, Pontes AL, Andrade SG. 27. Comportamento das cepas Y e Peruana do Trypanosoma cruzi, após passagens em diferentes meios. Memórias do Instituto Oswaldo Cruz, 80:41-50, 1985.
22. McDaniel JP, Dvorak JA. Identification, isolation and characterization of naturally occurring Trypanosoma cruzi variants. Molecular and Biochemical Parasitology 57:213-222, 1993.
23. Miles MA, Apt W, Widmer G, Povoa MM, Schofield C J. Isoenzyme heterogeneity and numerical taxonomy of Trypanosoma cruzi stocks from Chile. Transactions of the Royal Society of Tropical Medicine and Hygiene 78:526-535, 1984.
24. Miles MA, Lanham SM, Souza AA, Povoa M. Further enzymic characters of Trypanosoma cruzi and their evaluation for strain identification. Transactions of the Royal Society of Tropical Medicine and Hygiene 74:221-237, 1980.
25. Miles MA, ToyePJ, Sarah C, Oswaldo CC, Godfrey DG. The identification by isoenzyme patterns of two distinct strain groups of Trypanosoma cruzi, circulating independently in a rural area of Brazil. Transactions of the Royal Society of Tropical Medicine and Hygiene 71:217-225, 1977.
26. Morel C, Chiari E, Camargo EP, Mattei DM, Romanha AJ, Simpson L. Strains and clones of Trypanosoma cruzi can be characterized by pattern of restriction endonuclease products of kinetoplast DNA minicircles. Proceedings of the National Academy of Sciences USA 67:6810-6814, 1980.
27. Nussenzweig VB, Goble F. Further studies on the antigenic constitution of strains of Trypanosoma (Schizotrypanum) cruzi Experimental Parasitology 18:224-230, 1966.
28. Postan M, Dvorak JM, McDaniel JP. Studies of Trypanosoma cruzi clones in inbred mice. I. A comparison of the course of infection of C3H/HEN mice with two clones isolated from a common source. The American Journal o f Tropical Medicine and Hygiene 32:497-506, 1983.
29. Saravia NG, Holguin AF, Cibulskis RE, D'Alessandro A. Divergent isoenzyme profiles of sylvatic and domiciliary Trypanosoma cruzi in the highlands of Colombia. The American Journal of Tropical Medicine and Hygiene 36:59-69, 1987.
30. Tibayrenc M, Echalar L, Dujardin JP, Poch O, Desjeux P. The microdistribution of isoenzymic strains of Trypanosoma cruzi in southern Bolivia; new isoenzyme profiles and furtherarguments against Mendciian sexuality. Transactions of the Royal Society of Tropical Medicine and Hygiene 78:519- 525, 1984.
31. Tibayrenc M, Ward P, Moya A, Ayala JF. Natural populations of Trypanosoma cruzi, the agent of Chagas'disease. Proceedings of the National Academy of Sciences USA 83:115-119, 1986.
32. Widmer G, Marinkelle CJ, Guhl F, Miles MA. Isozyme profiles of Trypanosoma cruzi stocks from Colombia and Salvador. Annals ofTropical Medicine and Parasitology 3:253-257, 1985.

Adress to correspondence:

Dr

^a Sonia G. Andrade.

Centro de Pesquisas Gonçalo Moniz/FIOCRUZ/UFBA.

R. Valdemar Falcão 121 Brotas,

40295-001

Salvador, BA, Brasil.

Publication Dates

Publication in this collection
10 Apr 2013
Date of issue
Dec 1994

History

Accepted
25 Mar 1994
Received
25 Mar 1994

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

[1] 1. Andrade SG. Caracterização de cepas do Trypanosoma cruzi isoladas do Recôncavo Bahiano. Revista de Patologia Tropical 3:65-121, 1974.

[2] 2. Andrade SG. Morphological and behavioral characterization of Trypanosoma cruzi strains. Revista da Sociedade Brasileira de Medicina Tropical 18(suppl):39-46, 1985.

[3] 3. Andrade SG, Andrade, ZA, Figueira RM. Estudo experimental sobre a resistência de uma cepa do Trypanosoma cruzi ao Bay 2502. Revistado Instituto de Medicina Tropical de São Paulo 19:124-129,1977.

[4] 4. Andrade SG, Figueira RM. Estudo experimental sobre a ação terapêutica da droga R07-1051 na infeção por diferentes cepas do Trypanosoma cruzi Revista do Instituto de Medicina Tropical de São Paulo 9:335-341, 1977.

[5] 5. Andrade SG, Figueira RM, Carvalho ML, Gorini DF. Influência da cepa do Trypanosoma cruzi na resposta à terapêutica pelo Bay 2502 (Resultados de tratamento a longo prazo). Revista do Instituto de Medicina Tropical de São Paulo 17:380-389, 1975.

[6] 6. Andrade SG, Magalhães JB, Pontes AL. Evaluation of chemotherapy with benznidazole and nifurtimox in mice infected with Trypanosoma cruzi strains of different types. Bulletin of the World Health Organization 63:721-726, 1985.

[7] 7. Andrade V, Brodskyn C, Andrade SG. Correlation between isoenzymepatterns andbiologicalbehavior of different strains of Trypanosoma cruzi Transactions of the Royal Society of Tropical Medicine and Hygiene 77:796-799, 1983.

[8] 8. Bogliolo AR, Chiari E, Silva-Pereira RO, Silva Pereira AA. A comparative study of Trypanosoma cruzi enzyme polymorphism in South America. Brazilian JoumalofMedical and Bio logical Research 19: 673-683, 1986.

[9] 9. Bogliolo AR, Lanham SM, Corredor A. Further enzyme polymorphism in Trypanosoma cruzi from Colombia. Brazilian Journal of Medical and Biological Research 18: 427-433, 1985.

[10] 10. Brener Z, Costa CAG, Chiari C. Differences in the susceptibility of Trypanosoma cruzi strains to active chemotherapeutic agents. Revista do Instituto de Medicina Tropical de São Paulo 18:450-455, 1976.

[11] 11. Bongertz V, Dvorak JA. Trypanosoma cruzi: antigenic analysis of cloned stocks. The American Journal of Tropical Medicine and Hygiene 32:716- 722, 1983.

[12] 12. Carneiro M, Chiari E, Gonçalves AM, Silva Pereira AA, Morel CM, Romanha AJ. Changes in the isoenzyme and kinetoplast DNA patterns of Trypanosoma cruzi strains induced by maintenance in mice. Acta Tropica 47:35-45, 1990.

[13] 13. Castro Silva R, Santiago CMG, AndradeSG. Estudo da estabilidade dos caracteres isoenzimáticos de cepas do Trypanosoma cruzi em extratos enzimáticos estocados em nitrogênio líquido, por período prolongado. Memórias do Instituto Oswaldo Cruz 83(supl I):82, 1988.

[14] 14. Castro Silva R, Santiago CMG, Pontes AL, Andrade SG. Padrão isoenzimático da cepa Y do Trypanosoma cruzi após quimioterapia específica. Memórias do Instituto Oswaldo Cruz 84: 81-86, 1989.

[15] 15. Chapman MD, Baggaley RC, Godfrey-Fausset P, Malpas TJ, White G, Canese J, Miles MAS. Trypanosoma cruzi from the Paraguayan chaco: isoenzyme profiles of strains isolated at Makthlawaiya. Journal of Protozoology 31:482- 486, 1984.

[16] 16. Dvorak JA, Hartman DL, Miles MA. Trypanosoma cruzi: Correlation of growth kinetics to zymodeme type in clones derived from various sources. Joumal of Protozoology 27:472-474, 1980.

[17] 17. FedericiEE, AbelmannWB,NevaFA. Chronicand progressive myocarditis in C3H mice infected with Trypanosoma cruzi The American Journal of Tropical Medicine and Hygiene 13:272-280, 1964.

[18] 18. Godfrey DG, Kilgour V. Enzyme electrophoresis in characterizing the causative organism of gambian trypanosomiasis. Transactions of the Royal Society ofTropicalMedicineand Hygiene 3:219-224,1976.

[19] 19. Goldberg SS , Silva Pereira AA. Enzyme variation among clones of Trypanosoma cruzi Journal of Parasitology 69:91-96, 1983.

[20] 20. Magalhães JB, Andrade SG. Estudo do comportamento de cepas de Trypanosoma cruzi após passagem em diferentes espécies de triatomíneos. Revista da Sociedade Brasileira de Medicina Tropical 24:209-216, 1991.

[21] 21. Magalhães JB, Pontes AL, Andrade SG. 27. Comportamento das cepas Y e Peruana do Trypanosoma cruzi, após passagens em diferentes meios. Memórias do Instituto Oswaldo Cruz, 80:41-50, 1985.

[22] 22. McDaniel JP, Dvorak JA. Identification, isolation and characterization of naturally occurring Trypanosoma cruzi variants. Molecular and Biochemical Parasitology 57:213-222, 1993.

[23] 23. Miles MA, Apt W, Widmer G, Povoa MM, Schofield C J. Isoenzyme heterogeneity and numerical taxonomy of Trypanosoma cruzi stocks from Chile. Transactions of the Royal Society of Tropical Medicine and Hygiene 78:526-535, 1984.

[24] 24. Miles MA, Lanham SM, Souza AA, Povoa M. Further enzymic characters of Trypanosoma cruzi and their evaluation for strain identification. Transactions of the Royal Society of Tropical Medicine and Hygiene 74:221-237, 1980.

[25] 25. Miles MA, ToyePJ, Sarah C, Oswaldo CC, Godfrey DG. The identification by isoenzyme patterns of two distinct strain groups of Trypanosoma cruzi, circulating independently in a rural area of Brazil. Transactions of the Royal Society of Tropical Medicine and Hygiene 71:217-225, 1977.

[26] 26. Morel C, Chiari E, Camargo EP, Mattei DM, Romanha AJ, Simpson L. Strains and clones of Trypanosoma cruzi can be characterized by pattern of restriction endonuclease products of kinetoplast DNA minicircles. Proceedings of the National Academy of Sciences USA 67:6810-6814, 1980.

[27] 27. Nussenzweig VB, Goble F. Further studies on the antigenic constitution of strains of Trypanosoma (Schizotrypanum) cruzi Experimental Parasitology 18:224-230, 1966.

[28] 28. Postan M, Dvorak JM, McDaniel JP. Studies of Trypanosoma cruzi clones in inbred mice. I. A comparison of the course of infection of C3H/HEN mice with two clones isolated from a common source. The American Journal o f Tropical Medicine and Hygiene 32:497-506, 1983.

[29] 29. Saravia NG, Holguin AF, Cibulskis RE, D'Alessandro A. Divergent isoenzyme profiles of sylvatic and domiciliary Trypanosoma cruzi in the highlands of Colombia. The American Journal of Tropical Medicine and Hygiene 36:59-69, 1987.

[30] 30. Tibayrenc M, Echalar L, Dujardin JP, Poch O, Desjeux P. The microdistribution of isoenzymic strains of Trypanosoma cruzi in southern Bolivia; new isoenzyme profiles and furtherarguments against Mendciian sexuality. Transactions of the Royal Society of Tropical Medicine and Hygiene 78:519- 525, 1984.

[31] 31. Tibayrenc M, Ward P, Moya A, Ayala JF. Natural populations of Trypanosoma cruzi, the agent of Chagas'disease. Proceedings of the National Academy of Sciences USA 83:115-119, 1986.

[32] 32. Widmer G, Marinkelle CJ, Guhl F, Miles MA. Isozyme profiles of Trypanosoma cruzi stocks from Colombia and Salvador. Annals ofTropical Medicine and Parasitology 3:253-257, 1985.