Intestinal spirochetosis: first cases reported in Brazil and the use of immunohistochemistry as an aid in histopathological diagnosis

Brito, T. de; Sandoval, Marina P.; Silva, Ana G.; Saad, R. C.; Colaiacovo, W.

doi:10.1590/S0036-46651996000100009

Abstracts

Colonization of the colon and rectum by intestinal spirochetes is detected for the first time in Brazil in 4 of 282 (1.41%) patients who had undergone sigmoidoscopy and/or colonoscopy with a histopathological diagnosis of chronic non specific-colitis. This frequency is probably understimated, since surgically obtained specimens were not considered in the present study. Histopathological diagnosis was performed using routine stains like hematoxylin-eosin which showed the typical, of 3-µm thick hematoxyphilic fringe on the brush border of the surface epithelium, and by silver stains like the Warthin-Starry stain. Immunohistochemical procedures using two, polyclonal, primary antibodies, one against Treponema pallidum and the other against Leptospira interrogans serovar copenhageni serogroup Icterohaemorrhagiae cross-reacted with spirochetal antigen/s producing a marked contrast of the fringe over the colonic epithelium, preserving the spiral-shaped morphology of the parasite. In one case with marked diarrhea, immunohistochemistry detected spirochetal antigen/s within a cell in an intestinal crypt, thus demonstrating that the infection can be more widely disseminated than suspected using routine stains. Immunohistochemical procedures, thus, greatly facilitate the histological diagnosis of intestinal spirochetosis and may contribute to a better understanding of the pathogenesis of the disease. Transmission and scanning electron microscopy performed in one case showed that the spirochete closely resembled the species designated as Brachyspira aalborgi.

Intestinal Spirochetosis; Brachyspira aalborgi; Immunohistochemistry; Anti-leptospiral and anti-treponemal sera; Antigen detection

A colonização do colo e reto por espiroquetas intestinais foi detectada pela primeira vez no Brasil em 4 de 282 (1,41%) pacientes submetidos a sigmoidoscopia e/ou colonoscopia com o diagnóstico histopatológico de colite crônica inespecífica. A frequência é provavelmente menor que a real, urna vez que espécimens cirúrgicos não foram considerados no presente estudo. O diagnóstico histopatológico foi feito através de colorações de rotina, tais como a hematoxilina-eosina, que mostraram franja basofílica de 3µ de espessura sobre a borda em escova do epitélio e por impregnações pela prata como o método de Whartin-Starry. Métodos imunohistoquímicos, usando dois soros primários policlonais, um contra o Treponema pallidum e o outro contra Leptospira interrogans, serovar copenhageni serogrupo Icterohaemorrhagiae revelaram reatividade cruzada com os antígeno/s dos espiroquetas produzindo um contraste acentuado da franja sobre o epitélio colônico e preservando a morfologia espiralada do parasita. Em um paciente, com acentuada diarréia, a imunohistoquímica detectou antígenos de espiroqueta no citoplasma de uma célula da cripta intestinal, demonstrando que a infecção estava mais disseminada do que parecia pelas colorações comuns. Portanto, os métodos imunohistoquímicos facilitam o diagnóstico histológico da espiroquetose intestinal e podem , também, contribuir para a melhor compreensão da patogenia da doença. A microscopia eletrônica de transmissão e a microscopia de varredura, realizadas em um paciente, demonstraram que os espiroquetas lembram a espécie designada como Brachyspira aalborgi.

PATHOLOGY AND PATHOGENESIS

Intestinal spirochetosis: first cases reported in Brazil and the use of immunohistochemistry as an aid in histopathological diagnosis

Espiroquetose intestinal: primeiros casos no Brasil e o uso de técnicas imunohistoquímicas como ajuda no diagnóstico histopatológico

T. de Brito; Marina P. Sandoval; Ana G. Silva; R. C. Saad; W. Colaiacovo

Institute of Tropical Medicine and University Hospital, University of S. Paulo Medical School, S. Paulo, Brazil

^{Correspondence to} Correspondence to: Thales De Brito, M.D. Instituto de Medicina Tropical de S. Paulo (II), 2º andar, sala 5 Av. Dr. Enéas de Carvalho Aguiar, 470 05403-000 São Paulo, SP, Brazil

SUMMARY

Colonization of the colon and rectum by intestinal spirochetes is detected for the first time in Brazil in 4 of 282 (1.41%) patients who had undergone sigmoidoscopy and/or colonoscopy with a histopathological diagnosis of chronic non specific-colitis. This frequency is probably understimated, since surgically obtained specimens were not considered in the present study.

Histopathological diagnosis was performed using routine stains like hematoxylin-eosin which showed the typical, of 3-µm thick hematoxyphilic fringe on the brush border of the surface epithelium, and by silver stains like the Warthin-Starry stain. Immunohistochemical procedures using two, polyclonal, primary antibodies, one against Treponema pallidum and the other against Leptospira interrogans serovar copenhageni serogroup Icterohaemorrhagiae cross-reacted with spirochetal antigen/s producing a marked contrast of the fringe over the colonic epithelium, preserving the spiral-shaped morphology of the parasite. In one case with marked diarrhea, immunohistochemistry detected spirochetal antigen/s within a cell in an intestinal crypt, thus demonstrating that the infection can be more widely disseminated than suspected using routine stains. Immunohistochemical procedures, thus, greatly facilitate the histological diagnosis of intestinal spirochetosis and may contribute to a better understanding of the pathogenesis of the disease.

Transmission and scanning electron microscopy performed in one case showed that the spirochete closely resembled the species designated as Brachyspira aalborgi.

Keywords: Intestinal Spirochetosis, Brachyspira aalborgi, Immunohistochemistry, Anti-leptospiral and anti-treponemal sera, Antigen detection.

RESUMO

A colonização do colo e reto por espiroquetas intestinais foi detectada pela primeira vez no Brasil em 4 de 282 (1,41%) pacientes submetidos a sigmoidoscopia e/ou colonoscopia com o diagnóstico histopatológico de colite crônica inespecífica. A frequência é provavelmente menor que a real, urna vez que espécimens cirúrgicos não foram considerados no presente estudo.

O diagnóstico histopatológico foi feito através de colorações de rotina, tais como a hematoxilina-eosina, que mostraram franja basofílica de 3µ de espessura sobre a borda em escova do epitélio e por impregnações pela prata como o método de Whartin-Starry. Métodos imunohistoquímicos, usando dois soros primários policlonais, um contra o Treponema pallidum e o outro contra Leptospira interrogans, serovar copenhageni serogrupo Icterohaemorrhagiae revelaram reatividade cruzada com os antígeno/s dos espiroquetas produzindo um contraste acentuado da franja sobre o epitélio colônico e preservando a morfologia espiralada do parasita. Em um paciente, com acentuada diarréia, a imunohistoquímica detectou antígenos de espiroqueta no citoplasma de uma célula da cripta intestinal, demonstrando que a infecção estava mais disseminada do que parecia pelas colorações comuns. Portanto, os métodos imunohistoquímicos facilitam o diagnóstico histológico da espiroquetose intestinal e podem , também, contribuir para a melhor compreensão da patogenia da doença.

A microscopia eletrônica de transmissão e a microscopia de varredura, realizadas em um paciente, demonstraram que os espiroquetas lembram a espécie designada como Brachyspira aalborgi.

Full text available only in PDF format.

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ACKNOWLEDGEMENTS

Financial support for this project was provided by a grant from CNPq (300857/94-2) and by LIM-03 and LIM-06 "Hospital das Clínicas". We thank Miss Maria Elí P. de Castro for secretarial help.

Recebido para publicação em 29/01/1996

Aceito para publicação em 07/03/1996.

1. ANTONAKOPOULOS, G.; NEWMAN, J. & WILKINSON, M. - Intestinal spirochetosis: an electron microscopic study of an unusual case. Histopathology, 6: 477-488, 1982.
2. COOPER, C; COTTON, D.W.K.; HUDSON, M.J.; KIRKHAM, N. & WILMOTT, E.E.W. - Rectal spirochetosis in homosexual men: characterization of the organism and pathophysiology. Genitourin. Med, 62: 47-52, 1986.
3. COYNE, J.O.; CURRY, A.; PURNELL, P. & HABOUBI, N.T. - Colonic tubular adenomas and intestinal spirochaetosis: an incompatible association. Histopathology, 27: 377-379, 1995.
4. CRUCIOLI, V. & BUSUTTIL, A. - Human intestinal spirochetosis. Scand. J. Gastroent., 70: 177-179, 1981.
5. COTTON, D.W.K.; KIRKHAM, N. & HICKS, D.A. - Rectal spirochetosis. Brit. J. vener. Dis., 60: 106-109, 1984.
6. DAUZAN, Y.R.; MERLIO, J.P.H.; GRELIER, P. et al. - La spirochétose colo-rectale: une entité anatomo-pathologique? Ann. Path., 10: 258-261, 1990.
7. DELLADETSIMA, K.; MARKAKI, S. & PAPADIMITRIOU, K. - Intestinal spirochetosis. Light and electron microscopic study. Path. Res. Pract., 182: 780-782, 1987.
8. GAD, A.; WILLÉN, R.; FURUGARD, K.; FORS, B. & HRADSKY, M. - Intestinal spirochetosis as a cause of long standing diarrhea. Upsala J. med. Sci, 82: 49-54, 1977.
9. GOMES, J.M. & PESSOA, S.B. - Um caso de dyarrhea dysenteriforme grave provocada por espirochetas. An. paul. Med. Cirurg., 14: 167-169, 1926.
10. GUCCION, J.G.; BENATOR, D.A.; ZELLER, J.; TERMANINI, B. & SAINI, N. - Intestinal spirochetosis and acquired immunodeficiency syndrome: ultrastructural studies of two cases. Ultrastruct. Path., 19: 15-22, 1995.
11. HARLAND, W.A. & LEE, F.D. - Intestinal spirochetosis. Brit. med. J., 3: 718-719, 1967.
12. HOVIND-HOUGEN, K.; ANDERSEN, A.B.; NIELSEN, R.H. et al. - Intestinal spirochetosis: morphological characterization and cultivation of the spirochete Brachyspira aalborgi gen. nov., sp. nov. J. clin. Microbiol., 16: 1127-1136, 1982.
13. HSU, S.M. & FANGER, H. - Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase technique: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem., 29: 557-580, 1981.
14. LEE. F.D.; KRAZZEWSKI, A.; GORDON. J. et al. - Intestinal spirochetosis. Gut, 12: 126-133, 1971.
15. LEE, J.I. & HAMPSON, D.J. - Genetic characterization of intestinal spirochaetes and their association with disease. J. med. Microbiol., 40: 365-371, 1994.
16. LINDBOE, C.F.; TOSTRUP, N.E.; NERSUND, R. & REKKAVIK, G. - Human intestinal spirochaetosis in mid Norway. APMIS, 101: 858-864, 1993.
17. LINDBOE, C.F.; ENGESVOLL, I.; DARELL, M. & KOPSTAD, G. - Immunoglobulin containing cells in the colonic mucosa in patients with human intestinal spirochetosis. APMIS, 102: 849-854, 1994.
18. LINS, A. - Sobre o Spirochela eurygyruta e a dysenteria espirillar. Arch. bras. Med., 14: 420-426, 1924.
19. MAZZA, S. - Espiroquetosis apendiculares. Pren. méd. argent., 17: 464-468, 1930.
20. NATHWANI, D.; GREEN, S.T. & BOYD, J.D. - Intestinal spirochaetosis in a man with acquired immune deficiency syndrome (AIDS). J. Infect, 21: 318-319, 1990.
21. NIELSEN, R.H.; ORHOLM, M.; PEDERSEN, J.O. et al. - Coloretal spirochetosis: clinical significance of the infestation. Gastroenterology, 85: 62-67, 1983.
22. NIELSEN, R.H.; LUNDBECK, F.A.; TEGLBJAERG, P.S.; GINNERUP, P. & HOVIND-HOUGEN, K. - Intestinal spirochetosis of the vermiform appendix. Gastroenterology, 88: 971-977, 1985.
23. RODGERS, F.G.; RODGERS, C.; SHELTON, A.P. & HAWKEY, C.J. - Proposed pathogenic mechanism for the diarrhea associated with human intestinal spirochetes. Amer. J. clin. Path., 86: 679-682, 1986.
24. SURAWICZ, C.M.; ROBERTS, P.L.; ROMPALO, A. et al. - Intestinal spirochetosis in homosexual men. Amer. J. Med., 82: 587-592, 1987.
25. TAKEUCHI, A.; JERVIS, H.R.; NAKAZAWA, H. & ROBINSON, D.M. - Spiral-shaped organisms on the surface colonic epithelium of the monkey and men. Amer. J. clin. Nutr., 27: 1287-1296, 1974.
26. TEGLJAERG, P.S. - Intestinal spirochetosis. Curr. Top. Path., 81: 247-256, 1990.
27. WILLÉN, R.; CARLÉN, B.; CRONSTEDT, J. & WILLÉN, H. - Intestinal spirochetosis of the colon diagnosed with colono-ileoscopy and multiple biopsies. Endoscopy, 17: 86-88, 1985.

Correspondence to:

Thales De Brito, M.D.

Instituto de Medicina Tropical de S. Paulo (II), 2º andar, sala 5

Av. Dr. Enéas de Carvalho Aguiar, 470

05403-000 São Paulo, SP, Brazil

Publication Dates

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

History

Received
29 Jan 1996
Accepted
07 Mar 1996

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

[1] 1. ANTONAKOPOULOS, G.; NEWMAN, J. & WILKINSON, M. - Intestinal spirochetosis: an electron microscopic study of an unusual case. Histopathology, 6: 477-488, 1982.

[2] 2. COOPER, C; COTTON, D.W.K.; HUDSON, M.J.; KIRKHAM, N. & WILMOTT, E.E.W. - Rectal spirochetosis in homosexual men: characterization of the organism and pathophysiology. Genitourin. Med, 62: 47-52, 1986.

[3] 3. COYNE, J.O.; CURRY, A.; PURNELL, P. & HABOUBI, N.T. - Colonic tubular adenomas and intestinal spirochaetosis: an incompatible association. Histopathology, 27: 377-379, 1995.

[4] 4. CRUCIOLI, V. & BUSUTTIL, A. - Human intestinal spirochetosis. Scand. J. Gastroent., 70: 177-179, 1981.

[5] 5. COTTON, D.W.K.; KIRKHAM, N. & HICKS, D.A. - Rectal spirochetosis. Brit. J. vener. Dis., 60: 106-109, 1984.

[6] 6. DAUZAN, Y.R.; MERLIO, J.P.H.; GRELIER, P. et al. - La spirochétose colo-rectale: une entité anatomo-pathologique? Ann. Path., 10: 258-261, 1990.

[7] 7. DELLADETSIMA, K.; MARKAKI, S. & PAPADIMITRIOU, K. - Intestinal spirochetosis. Light and electron microscopic study. Path. Res. Pract., 182: 780-782, 1987.

[8] 8. GAD, A.; WILLÉN, R.; FURUGARD, K.; FORS, B. & HRADSKY, M. - Intestinal spirochetosis as a cause of long standing diarrhea. Upsala J. med. Sci, 82: 49-54, 1977.

[9] 9. GOMES, J.M. & PESSOA, S.B. - Um caso de dyarrhea dysenteriforme grave provocada por espirochetas. An. paul. Med. Cirurg., 14: 167-169, 1926.

[10] 10. GUCCION, J.G.; BENATOR, D.A.; ZELLER, J.; TERMANINI, B. & SAINI, N. - Intestinal spirochetosis and acquired immunodeficiency syndrome: ultrastructural studies of two cases. Ultrastruct. Path., 19: 15-22, 1995.

[11] 11. HARLAND, W.A. & LEE, F.D. - Intestinal spirochetosis. Brit. med. J., 3: 718-719, 1967.

[12] 12. HOVIND-HOUGEN, K.; ANDERSEN, A.B.; NIELSEN, R.H. et al. - Intestinal spirochetosis: morphological characterization and cultivation of the spirochete Brachyspira aalborgi gen. nov., sp. nov. J. clin. Microbiol., 16: 1127-1136, 1982.

[13] 13. HSU, S.M. & FANGER, H. - Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase technique: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem., 29: 557-580, 1981.

[14] 14. LEE. F.D.; KRAZZEWSKI, A.; GORDON. J. et al. - Intestinal spirochetosis. Gut, 12: 126-133, 1971.

[15] 15. LEE, J.I. & HAMPSON, D.J. - Genetic characterization of intestinal spirochaetes and their association with disease. J. med. Microbiol., 40: 365-371, 1994.

[16] 16. LINDBOE, C.F.; TOSTRUP, N.E.; NERSUND, R. & REKKAVIK, G. - Human intestinal spirochaetosis in mid Norway. APMIS, 101: 858-864, 1993.

[17] 17. LINDBOE, C.F.; ENGESVOLL, I.; DARELL, M. & KOPSTAD, G. - Immunoglobulin containing cells in the colonic mucosa in patients with human intestinal spirochetosis. APMIS, 102: 849-854, 1994.

[18] 18. LINS, A. - Sobre o Spirochela eurygyruta e a dysenteria espirillar. Arch. bras. Med., 14: 420-426, 1924.

[19] 19. MAZZA, S. - Espiroquetosis apendiculares. Pren. méd. argent., 17: 464-468, 1930.

[20] 20. NATHWANI, D.; GREEN, S.T. & BOYD, J.D. - Intestinal spirochaetosis in a man with acquired immune deficiency syndrome (AIDS). J. Infect, 21: 318-319, 1990.

[21] 21. NIELSEN, R.H.; ORHOLM, M.; PEDERSEN, J.O. et al. - Coloretal spirochetosis: clinical significance of the infestation. Gastroenterology, 85: 62-67, 1983.

[22] 22. NIELSEN, R.H.; LUNDBECK, F.A.; TEGLBJAERG, P.S.; GINNERUP, P. & HOVIND-HOUGEN, K. - Intestinal spirochetosis of the vermiform appendix. Gastroenterology, 88: 971-977, 1985.

[23] 23. RODGERS, F.G.; RODGERS, C.; SHELTON, A.P. & HAWKEY, C.J. - Proposed pathogenic mechanism for the diarrhea associated with human intestinal spirochetes. Amer. J. clin. Path., 86: 679-682, 1986.

[24] 24. SURAWICZ, C.M.; ROBERTS, P.L.; ROMPALO, A. et al. - Intestinal spirochetosis in homosexual men. Amer. J. Med., 82: 587-592, 1987.

[25] 25. TAKEUCHI, A.; JERVIS, H.R.; NAKAZAWA, H. & ROBINSON, D.M. - Spiral-shaped organisms on the surface colonic epithelium of the monkey and men. Amer. J. clin. Nutr., 27: 1287-1296, 1974.

[26] 26. TEGLJAERG, P.S. - Intestinal spirochetosis. Curr. Top. Path., 81: 247-256, 1990.

[27] 27. WILLÉN, R.; CARLÉN, B.; CRONSTEDT, J. & WILLÉN, H. - Intestinal spirochetosis of the colon diagnosed with colono-ileoscopy and multiple biopsies. Endoscopy, 17: 86-88, 1985.