Isoenzyme profile as parameter to differentiate pathogenic strains of Entamoeba histolytica in Brazil

Martinez, Ana Maria Barral de; Gomes, Maria Aparecida; Viana, João da Costa; Romanha, Alvaro José; Silva, Edward Felix

doi:10.1590/S0036-46651996000600004

Abstracts

The isoenzyme profiles (IP) of 33 strains of Entamoeba histolytica isolated from patients and carriers of two regions in Brazil (Amazonia and Southeast) were determined. The enzymes phosphoglucomutase, glucose-phosphate isomerase, hexokinase and malic enzyme were considered. IP of the strains was correlated with culture conditions, time of maintenance in laboratory and clinical history of patients. The strains were maintained under polyxenic, monoxenic and axenic culture conditions: 27 polyxenic, 1 polyxenic and monoxenic, 1 polyxenic, monoxenic and axenic and 4 axenic only. The patients were symptomatic and asymptomatic. The symptomatic patients presented either non dysenteric (NDC) or dysenteric colitis (DC), associated or not with hepatic abscess (HA). One patient presented anal amoeboma (AM). The analysis of IP for isolates maintained in polyxenic culture showed non pathogenic IP (I) for strains from carriers and patients with NDC, while the strains isolated from patients presenting DC, HA and AM resulted in isolates II or XIX pathogenic IP. This parameter was not able to differentiate strains from carriers from symptomatic patients when these strains were found in axenic or monoxenic culture. All these strains displayed pathogenic IP (II), demonstrating the inability of this parameter to classifying for virulence since it showed identical IP for strains isolated from carriers or symptomatic patients.

Entamoeba histolytica; Isoenzymatic profiles; Virulence; Diagnostic

O perfil isoenzimático (PI) de 33 cepas de E. histolytica, isoladas das regiões Amazônica e Sudeste do Brasil foi determinado. Foram consideradas as enzimas fosfoglicomutase, glicose fosfato isomerase, hexoquinase e enzima málica. O perfil obtido para cada cepa foi correlacionado com o meio e o tempo de manutenção em cultivo e com a história clínica do paciente. As cepas foram mantidas sob condições de cultivo axênico, monoxênico e polixênico: 27 polixênico, 1 polixênico e monoxênico, 1 polixênico, monoxênico e axênico e 4 somente em cultivo axênico. Os pacientes apresentaram sintomas ou não. Os pacientes sintomáticos apresentaram colite disentérica (CD) ou colite não disentérica (CND), associada ou não com abscesso hepático (AH). Um paciente apresentou ameboma anal (AM). A análise do PI das diferentes cepas mantidas em cultivo polixênico mostrou um PI não patogênico (I) para as cepas isoladas de portadores e pacientes com CND, enquanto as cepas isoladas de pacientes apresentando CD, AH ou AM resultou em isolados com PI patogênico (II e XIX). Analisando o PI das cepas mantidas em cultura axênica ou monoxênica, verificamos a inabilidade deste parâmetro para diferenciar cepas isoladas de portadores daquelas isoladas de pacientes sintomáticos. Todas estas cepas apresentaram PI patogênico, demonstrando a inabilidade deste parâmetro para classificar quanto à virulência.

PARASITOLOGY

Isoenzyme profile as parameter to differentiate pathogenic strains of Entamoeba histolytica in Brazil

Perfil isoenzimático como parâmetro para diferenciar cepas patogênicas de Entamoeba histolytica no Brasil

Ana Maria Barral de Martinez; Maria Aparecida Gomes; João da Costa Viana; Alvaro José Romanha; Edward Felix Silva

^{Correspondence to} Correspondence to: Dr. Edward Felix Silva Departamento de Parasitologia, ICB-UFMG Av. Antônio Carlos 6627, Pampulha 31270-901 Belo Horizonte, MG, Brazil

SUMMARY

The isoenzyme profiles (IP) of 33 strains of Entamoeba histolytica isolated from patients and carriers of two regions in Brazil (Amazonia and Southeast) were determined. The enzymes phosphoglucomutase, glucose-phosphate isomerase, hexokinase and malic enzyme were considered. IP of the strains was correlated with culture conditions, time of maintenance in laboratory and clinical history of patients. The strains were maintained under polyxenic, monoxenic and axenic culture conditions: 27 polyxenic, 1 polyxenic and monoxenic, 1 polyxenic, monoxenic and axenic and 4 axenic only. The patients were symptomatic and asymptomatic. The symptomatic patients presented either non dysenteric (NDC) or dysenteric colitis (DC), associated or not with hepatic abscess (HA). One patient presented anal amoeboma (AM). The analysis of IP for isolates maintained in polyxenic culture showed non pathogenic IP (I) for strains from carriers and patients with NDC, while the strains isolated from patients presenting DC, HA and AM resulted in isolates II or XIX pathogenic IP. This parameter was not able to differentiate strains from carriers from symptomatic patients when these strains were found in axenic or monoxenic culture. All these strains displayed pathogenic IP (II), demonstrating the inability of this parameter to classifying for virulence since it showed identical IP for strains isolated from carriers or symptomatic patients.

Keywords: Entamoeba histolytica; Isoenzymatic profiles; Virulence; Diagnostic.

RESUMO

O perfil isoenzimático (PI) de 33 cepas de E. histolytica, isoladas das regiões Amazônica e Sudeste do Brasil foi determinado. Foram consideradas as enzimas fosfoglicomutase, glicose fosfato isomerase, hexoquinase e enzima málica. O perfil obtido para cada cepa foi correlacionado com o meio e o tempo de manutenção em cultivo e com a história clínica do paciente. As cepas foram mantidas sob condições de cultivo axênico, monoxênico e polixênico: 27 polixênico, 1 polixênico e monoxênico, 1 polixênico, monoxênico e axênico e 4 somente em cultivo axênico. Os pacientes apresentaram sintomas ou não. Os pacientes sintomáticos apresentaram colite disentérica (CD) ou colite não disentérica (CND), associada ou não com abscesso hepático (AH). Um paciente apresentou ameboma anal (AM). A análise do PI das diferentes cepas mantidas em cultivo polixênico mostrou um PI não patogênico (I) para as cepas isoladas de portadores e pacientes com CND, enquanto as cepas isoladas de pacientes apresentando CD, AH ou AM resultou em isolados com PI patogênico (II e XIX). Analisando o PI das cepas mantidas em cultura axênica ou monoxênica, verificamos a inabilidade deste parâmetro para diferenciar cepas isoladas de portadores daquelas isoladas de pacientes sintomáticos. Todas estas cepas apresentaram PI patogênico, demonstrando a inabilidade deste parâmetro para classificar quanto à virulência.

Full text available only in PDF format.

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ACKNOWLEDGEMENTS

To FAPEMIG, FINEP and CNPq for supporting this work.

Recebido para publicação em 01/09/1995

Aceito para publicação em 04/12/1996

1. ANDREWS, B.; MENTZONI, L. & BJORVATN, L. - Zymodemes conversion of isolates of Entamoeba histolytica. Trans. roy. Soc. trop. Med. Hyg., 84: 63-65, 1990.
2. BLANC, D. - Determination of taxonomic status of pathogenic and nonpathogenic Entamoeba histolytica zymodemes using isoenzyme analysis. J. Protozool., 39: 471-479, 1992.
3. CLARK, C.G. & DIAMOND, L.S. - Pathogenicity, virulence and Entamoeba histolytica. Parasit. today, 10: 46-47, 1994.
4. CRUZ REYES, J.; SPICE, W.; REHMAN, T.; GISBORNE, E. & ACKERS, J. - Ribosomal DNA sequences in the differentiation of pathogenic and nonpathogenic isolates of Entamoeba histolytica. Parasitology, 104: 239-246, 1992.
5. CUNHA, A.S. - Patogenia da amebíase. Belo Horizonte, 1975. (Tese para Professor Titular - Faculdade de Medicina da UFMG).
6. CUNHA, A.S.; SILVA, E.F.; RASO, P. & MELO, S.M. - Patogenia da amebíase. I. Aspectos clínicos da amebíase no Brasil. Estudo realizado em tręs grupos populacionais de tręs regiőes geográficas distintas. Rev. Inst. Med. trop. S. Paulo, 19: 289-300, 1977.
7. FARRI, T.; SARGEAUNT, P.; WARHURST, D. & WILLIAMS, J. - Electrophoretic isoenzyme patterns of the pathogenic and nonpathogenic intestinal amoebae of man. Trans. roy. Soc. trop. Med. Hyg., 73: 225-227, 1979.
8. GATHIRAM, V. & JACKSON, T. - Frequency distribution Entamoeba histolytica in a rural South African population. Lancet, 30: 719-721, 1985.
9. GATHIRAM, V. & JACKSON, T. - Pathogenic zymodemes of Entamoeba histolytica remain unchanged throughout their life-cycle. Trans. roy. Soc. trop. Med. Hyg., 84: 806-807, 1990.
10. MIRELMAN, D.; BRACHA, R.; WEXLER, A. & CHAYEN, A. - Alteration of isoenzyme patterns of a cloned culture of non-pathogenic Entamoeba histolytica upon changes in growth conditions. Arch. Invest. méd. (Méx.), 17: 187-193, 1986.
11. MIRELMAN, D. - Effects of culture conditions and bacterial associates on the zymodeme of Entamoeba histolytica. Parasit. today, 3: 37-40, 1987.
12. NOZAKI, T.; MOTTA, S.R.N.; KOBAYNOZAKI, T. et al. - Zymodemes of Entamoeba histolytica isolated in the Amazon and the north-east of Brazil. Trans. roy. Soc. trop. Med. Hyg., 84: 387-388, 1990.
13. OROZCO, E.; GUARNEROS, G. & MARTINEZ-PALOMO, A. - Entamoeba histolytica: phagocytosis as a virulence factor. J. exp. Med., 158: 1511-1521, 1983.
14. ROBINSON, G. - The laboratory diagnosis of human amoebae. Trans. roy. Soc. trop. Med. Hyg., 62: 285-294, 1968.
15. SARGEAUNT, P. & WILLIAMS, J. - Electrophoretic isoenzyme of Entamoeba histolytica and Entamoeba coli. Trans. roy. Soc. trop. Med. Hyg., 72: 164-166, 1978a.
16. SARGEAUNT, P. & WILLIAMS, J. - The differentiation of invasive and non-invasive Entamoeba histolytica by isoenzyme electrophoresis. Trans. roy. Soc. trop. Med. Hyg., 72: 519-521, 1978b.
17. SARGEAUNT, P.; BAVEJA, U.; NANDA, R. & ANAND, B. - Influence of geographical factors in the distribution of pathogenic zymodemes of Entamoeba histolytica. Identification of Zymodeme XIV in India. Trans. roy. Soc. trop. Med. Hyg., 78: 96-101, 1984.
18. SARGEAUNT, P. - Zymodemes expressing possible genetic exchange in Entamoeba histolytica. Trans. roy. Soc. trop. Med. Hyg., 79: 86-89, 1985.
19. SARGEAUNT, P. - The reliability of Entamoeba histolytica zymodemes in clinical diagnosis. Parasit. today, 3: 40-43, 1987.
20. SARGEAUNT, P. - A survey of Entamoeba histolytica and Entamoeba dispar (Brumpt). Infections on Mahé, the Seychelles. Arch. med. Res., 23: 265-267, 1992.
21. SILVA, E.F. - Estudos sobre a Entamoeba moshkosvskii. II. Novos focos em diversos tipos de coleçőes hídricas no Brasil e no Uruguai. Rev. Inst. Med. trop. S. Paulo, 16: 203-221, 1974.
22. SILVA, E.F.; SILVA PEREIRA, A.A. & BOGLIOLO, A.R. - Isoenzymes patterns of Entamoeba histolytica stocks grown in axenic and non-axenic conditions. Mem. Inst. Oswaldo Cruz, 83(Suppl. 1): 259, 1988.
23. STRACHAN, W.; CHIODINI, P.; SPICE, W.; MOODY, A. & ACKERS, J. - Immunological differentiation of pathogenic and nonpathogenic isolates of Entamoeba histolytica. Lancet, 12: 561-563, 1988.
24. TANNICH, E. & BURCHARD, G. - Differentiation of pathogenic from non pathogenic Entamoeba histolytica by restriction fragment analysis of a single gene amplified in vitro. J. clin. Microbiol., 29: 250-255, 1991.
25. TRISSL, D.; MARTINEZ-PALOMO, A.; DE LA TORRE, M.; DE LA HOZ, R. & PEREZ SUAREZ, E. - Surfaces properties of Entamoeba. Increased rate of human erythrocyte phagocytosis in pathogenic strains. J.exp. Med., 148: 1137-1145, 1978.
26. VARGAS, M.A. & OROZCO, E. - Entamoeba histolytica. Changes in the zymodeme of cloned nonpathogenic trophozoites cultured under differents conditions. Parasit. Res., 79: 353-357, 1993.
27. WALSH, J. - Amebiasis in the world. Arch. Invest. méd. (Méx.), 17: 385-389, 1986.
28. WHO - Amebiasis. Wld. Hlth. Org. techn. Rep. Ser., (421), 1969.
²⁹
WHO informal meeting on strategies for control of amebiase. Geneve, 1984.

Correspondence to:

Dr. Edward Felix Silva

Departamento de Parasitologia, ICB-UFMG

Av. Antônio Carlos 6627, Pampulha

31270-901 Belo Horizonte, MG, Brazil

Publication Dates

Publication in this collection
22 Sept 2006
Date of issue
Dec 1996

History

Accepted
04 Dec 1996
Received
01 Sept 1995

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

[1] 1. ANDREWS, B.; MENTZONI, L. & BJORVATN, L. - Zymodemes conversion of isolates of Entamoeba histolytica. Trans. roy. Soc. trop. Med. Hyg., 84: 63-65, 1990.

[2] 2. BLANC, D. - Determination of taxonomic status of pathogenic and nonpathogenic Entamoeba histolytica zymodemes using isoenzyme analysis. J. Protozool., 39: 471-479, 1992.

[3] 3. CLARK, C.G. & DIAMOND, L.S. - Pathogenicity, virulence and Entamoeba histolytica. Parasit. today, 10: 46-47, 1994.

[4] 4. CRUZ REYES, J.; SPICE, W.; REHMAN, T.; GISBORNE, E. & ACKERS, J. - Ribosomal DNA sequences in the differentiation of pathogenic and nonpathogenic isolates of Entamoeba histolytica. Parasitology, 104: 239-246, 1992.

[5] 5. CUNHA, A.S. - Patogenia da amebíase. Belo Horizonte, 1975. (Tese para Professor Titular - Faculdade de Medicina da UFMG).

[6] 6. CUNHA, A.S.; SILVA, E.F.; RASO, P. & MELO, S.M. - Patogenia da amebíase. I. Aspectos clínicos da amebíase no Brasil. Estudo realizado em tręs grupos populacionais de tręs regiőes geográficas distintas. Rev. Inst. Med. trop. S. Paulo, 19: 289-300, 1977.

[7] 7. FARRI, T.; SARGEAUNT, P.; WARHURST, D. & WILLIAMS, J. - Electrophoretic isoenzyme patterns of the pathogenic and nonpathogenic intestinal amoebae of man. Trans. roy. Soc. trop. Med. Hyg., 73: 225-227, 1979.

[8] 8. GATHIRAM, V. & JACKSON, T. - Frequency distribution Entamoeba histolytica in a rural South African population. Lancet, 30: 719-721, 1985.

[9] 9. GATHIRAM, V. & JACKSON, T. - Pathogenic zymodemes of Entamoeba histolytica remain unchanged throughout their life-cycle. Trans. roy. Soc. trop. Med. Hyg., 84: 806-807, 1990.

[10] 10. MIRELMAN, D.; BRACHA, R.; WEXLER, A. & CHAYEN, A. - Alteration of isoenzyme patterns of a cloned culture of non-pathogenic Entamoeba histolytica upon changes in growth conditions. Arch. Invest. méd. (Méx.), 17: 187-193, 1986.

[11] 11. MIRELMAN, D. - Effects of culture conditions and bacterial associates on the zymodeme of Entamoeba histolytica. Parasit. today, 3: 37-40, 1987.

[12] 12. NOZAKI, T.; MOTTA, S.R.N.; KOBAYNOZAKI, T. et al. - Zymodemes of Entamoeba histolytica isolated in the Amazon and the north-east of Brazil. Trans. roy. Soc. trop. Med. Hyg., 84: 387-388, 1990.

[13] 13. OROZCO, E.; GUARNEROS, G. & MARTINEZ-PALOMO, A. - Entamoeba histolytica: phagocytosis as a virulence factor. J. exp. Med., 158: 1511-1521, 1983.

[14] 14. ROBINSON, G. - The laboratory diagnosis of human amoebae. Trans. roy. Soc. trop. Med. Hyg., 62: 285-294, 1968.

[15] 15. SARGEAUNT, P. & WILLIAMS, J. - Electrophoretic isoenzyme of Entamoeba histolytica and Entamoeba coli. Trans. roy. Soc. trop. Med. Hyg., 72: 164-166, 1978a.

[16] 16. SARGEAUNT, P. & WILLIAMS, J. - The differentiation of invasive and non-invasive Entamoeba histolytica by isoenzyme electrophoresis. Trans. roy. Soc. trop. Med. Hyg., 72: 519-521, 1978b.

[17] 17. SARGEAUNT, P.; BAVEJA, U.; NANDA, R. & ANAND, B. - Influence of geographical factors in the distribution of pathogenic zymodemes of Entamoeba histolytica. Identification of Zymodeme XIV in India. Trans. roy. Soc. trop. Med. Hyg., 78: 96-101, 1984.

[18] 18. SARGEAUNT, P. - Zymodemes expressing possible genetic exchange in Entamoeba histolytica. Trans. roy. Soc. trop. Med. Hyg., 79: 86-89, 1985.

[19] 19. SARGEAUNT, P. - The reliability of Entamoeba histolytica zymodemes in clinical diagnosis. Parasit. today, 3: 40-43, 1987.

[20] 20. SARGEAUNT, P. - A survey of Entamoeba histolytica and Entamoeba dispar (Brumpt). Infections on Mahé, the Seychelles. Arch. med. Res., 23: 265-267, 1992.

[21] 21. SILVA, E.F. - Estudos sobre a Entamoeba moshkosvskii. II. Novos focos em diversos tipos de coleçőes hídricas no Brasil e no Uruguai. Rev. Inst. Med. trop. S. Paulo, 16: 203-221, 1974.

[22] 22. SILVA, E.F.; SILVA PEREIRA, A.A. & BOGLIOLO, A.R. - Isoenzymes patterns of Entamoeba histolytica stocks grown in axenic and non-axenic conditions. Mem. Inst. Oswaldo Cruz, 83(Suppl. 1): 259, 1988.

[23] 23. STRACHAN, W.; CHIODINI, P.; SPICE, W.; MOODY, A. & ACKERS, J. - Immunological differentiation of pathogenic and nonpathogenic isolates of Entamoeba histolytica. Lancet, 12: 561-563, 1988.

[24] 24. TANNICH, E. & BURCHARD, G. - Differentiation of pathogenic from non pathogenic Entamoeba histolytica by restriction fragment analysis of a single gene amplified in vitro. J. clin. Microbiol., 29: 250-255, 1991.

[25] 25. TRISSL, D.; MARTINEZ-PALOMO, A.; DE LA TORRE, M.; DE LA HOZ, R. & PEREZ SUAREZ, E. - Surfaces properties of Entamoeba. Increased rate of human erythrocyte phagocytosis in pathogenic strains. J.exp. Med., 148: 1137-1145, 1978.

[26] 26. VARGAS, M.A. & OROZCO, E. - Entamoeba histolytica. Changes in the zymodeme of cloned nonpathogenic trophozoites cultured under differents conditions. Parasit. Res., 79: 353-357, 1993.

[27] 27. WALSH, J. - Amebiasis in the world. Arch. Invest. méd. (Méx.), 17: 385-389, 1986.

[28] 28. WHO - Amebiasis. Wld. Hlth. Org. techn. Rep. Ser., (421), 1969.

[29] ²⁹
WHO informal meeting on strategies for control of amebiase. Geneve, 1984.

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