El cultivo "in vitro" como instrumento práctico para el diagnóstico y aislamiento primario de Leishmania braziliensis braziliensis. 2. Estudios en pacientes de áreas endémicas

Cuba Cuba, C. A.; Netto, E. M.; Costa, J. L. M.; Barreto, A. C.; Marsden, P. D.

doi:10.1590/S0036-46651986000500006

Resúmenes

El cultivo "in vitro" de Leishmania braziliensis braziliensis constituye un método útil en el trabajo de campo, para el aislamiento primario de ésta subes-pécie de Leishmania. Cultivos en dos medios difásicos de agar sangre (DAB y EVANS) y dos medios líquidos (SCHNEIDER'S y AR-103) realizados en pacientes con lesiones cutáneas de Leishmaniasis Tegumentaria Americana (LTA) demostraron: 1) Similar sensibilidad de los medios DAB y Schneider's cuando utilizamos el procedimiento de aspiración de las muestras con aguja. 2) Rendimiento sensible y reproducible, con el medio DAB, cuando comparado, en repetidas ocasiones, con el medio EVANS. 3) Incremento significativo en el aislamiento primario de Leishmania braziliensis brazilensis mediante la ejecución, en la misma lesión de cada paciente, de tres aspiraciones consecutivas en sitios diferentes de la úlcera activa (50% de positividad, con el medio DAB).

Leishmaniose tegumentar; Leishmania braziliensis braziliensis; Diagnóstico pelo cultivo in vitro

In vitro cultivation of Lbb is a useful method in the field for primary isolation of this leismanial subspecies. Cultures in biphasic blood agar media (DAB and Evans) and liquid media (Schneiders and AR 103) done in patients with cutaneous leismanias showed, 1. A similar sensibility in DAB and Schneiders medium when needle aspirates were used to seed the cultures; 2. That DAB medium performed well when compared "with Evans medium; 3. A rise in isolation of Lbb if 3 aspirations taken from different sites in the same lesion were cultivated (50% positivity with DAB medium).

ARTIGOS ORIGINAIS

El cultivo "in vitro" como instrumento práctico para el diagnóstico y aislamiento primario de Leishmania braziliensis braziliensis. 2. Estudios en pacientes de áreas endémicas^* * Trabajo parcialmente financiado por CNPq (Pro. 403682/82) y OMS/TDR-RE (820526)

The viability of the "in vitro" cultivation procedure as a practical tool for diagnosis and primary isolation of Leishmania braziliensis braziliensis. 2. A study of patients from endemic areas

C. A. Cuba Cuba; E. M. Netto; J. L. M. Costa; A. C. Barreto; P. D. Marsden

Núcleo de Medicina Tropical e Nutrição, Universidade de Brasília, Caixa Postal 153121, 70.910 Brasília, D.F., Brasil

RESUMEN

El cultivo "in vitro" de Leishmania braziliensis braziliensis constituye un método útil en el trabajo de campo, para el aislamiento primario de ésta subes-pécie de Leishmania. Cultivos en dos medios difásicos de agar sangre (DAB y EVANS) y dos medios líquidos (SCHNEIDER'S y AR-103) realizados en pacientes con lesiones cutáneas de Leishmaniasis Tegumentaria Americana (LTA) demostraron: 1) Similar sensibilidad de los medios DAB y Schneider's cuando utilizamos el procedimiento de aspiración de las muestras con aguja. 2) Rendimiento sensible y reproducible, con el medio DAB, cuando comparado, en repetidas ocasiones, con el medio EVANS. 3) Incremento significativo en el aislamiento primario de Leishmania braziliensis brazilensis mediante la ejecución, en la misma lesión de cada paciente, de tres aspiraciones consecutivas en sitios diferentes de la úlcera activa (50% de positividad, con el medio DAB).

Unitermos: Leishmaniose tegumentar Leishmania braziliensis braziliensis Diagnóstico pelo cultivo in vitro.

SUMMARY

In vitro cultivation of Lbb is a useful method in the field for primary isolation of this leismanial subspecies. Cultures in biphasic blood agar media (DAB and Evans) and liquid media (Schneiders and AR 103) done in patients with cutaneous leismanias showed,

1. A similar sensibility in DAB and Schneiders medium when needle aspirates were used to seed the cultures;

2. That DAB medium performed well when compared "with Evans medium;

3. A rise in isolation of Lbb if 3 aspirations taken from different sites in the same lesion were cultivated (50% positivity with DAB medium).

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4. CHANG, K. P. & FISH, W. R. Leishmania In: JENSEN. J. B., ed. In vitro cultivation protozoa pathogenic to men and domestic animals. Boca Raton, Florida. CRC Press, 1981. p. 2-91.

Recebido para publicação em 12/11/1985.

1. AZEVEDO, H. F. & ROITMAN, I. Cultivation of Trypanosoma cruzi in defined media. In: MOREL, C M., ed. Genes and antigens parasites. A laboratory manual. 2nd. ed. Rio de Janeiro, Fundação Oswaldo Cruz, ,1984. p. 29-36.
2. BARKER, D. C. & BUTCHER, J. The use of PNA probes in the identification of leishmanias: discrimination between isolates of the Leishmania mexicana and L. braziliensis complex. Trans. ray. Soc. trop. med. Hyg., 77: 285-291, 1983.
3. BARRETO, A. C; CUBA CUBA, C. A.; MARSDEN, P. D.; VEXANAT, J. & BELDER, M. Características epidiemiológicas de Leishmaniose Tegumentar Americana em uma região endêmica do Estado da Bahia. Brasil. 1. Leishmaniose humana. Bol. Ofic. sanit. panamer., 90: 415-424, 1981.
5. CUBA, C. A.; VEXENAT, A. J.; LLANOS, E. A.; MARSDEN. P. D.; BARRETO. A. C. & ROSA, A. C. Experimental infections of wild caught specimens of Lutzomyia (N) whitmani (Díptera. Psychodidae) and their use for Leishmania identification. In: REUNIÃO ANUAL SOBRE PESQUISA BÁSICA EM DOENÇA DE CHAGAS, 9., Caxambu, 1982. Programa e resumos. Belo Horizonte, CNPq: FINEP, 1982. p. 149.
6 CUBA CUBA. C. A.; NETTO, M. E.: COSTA, L. J.; ROSA, A. de C ; BARRETO, A. C. & MARSDEN, P. D. Cutaneous leishmaniasis caused by Leishmania braziliensis braziliensis: Improving diagnosis by primary isolation in culture. In: REUNIÃO ANUAL SOBRE PESQUISA BÁSICA EM DOENÇA DE CHAGAS, li.. Caxambu, 1984. Programa e resumos. Ribeirão Preto. PIDE: CNPq. 1984. p. 129.
7 CUBA CUBA., C. A.; MILES. M. A.; VEXENAT, J. A.; Mc MaHON PRATT. D.: BARKER, D. C: BUTCHER, J. D.; BARRETO, A. C & MARSDEN, P. D. A forus of mucocutaneous leishhmaniasis in Três Braços, Bahia, Brazil. Characterization and identification of Leishmania stocks isolated from man and dogs. Trans. roy. Soc. trop. Med. Hyg., 79: 500-507, 1985.
8. EVANS, D. A.; LANHAM, S. M.; BALDWIN, C. I. & PETERS, W. Tne isolation and isoenzyme characterization of Leishmania braziliensis sbsp. from patients with cutaneous leishmaniasis acquired in Belize. Trans. roy. Soc. trop. Med. Hyg., 78: 35-42, 1984.
9. HENDRICKS, L. D. & WRIGHT, N. Diagnosis of cutaneous leishmaniasis by "in vitro" cultivation of saline aspirate in Schneider's Drosophila medium. Amer. J. trop. Med. Hyg., 28: 962-964, 1979.
10. HENDRICKS, L. D. & CHILDS, G. E. Present knowledge of the in vitro cultivation of Leishmania. In: THE IN VITEO CULTIVATION OF THE PATHOGENS OF TROPICAL DISEASES. Basel, Schwabe & Co. AG, 1980. p. 251-272. (Tropical Diseases Research Series No. 3).
11. LAINSON, R. & SHAW, J. J. Some problems in studies on parasites of the Leishmania braziliensis complex. In: Ecologie des Leishmanioses. Colloquies Internationaux du CNRS, n.° 239. Montpellier, 1977. p. 83-86.
12. LAINSON, R. & SHAW, J. J. The role of animals In the epidemiology of South American Leishmaniasis. In: LUMSDEN, W. H. R. & EVANS, D. A., ed. -Biology of the kinetoplastida. London, Academic Press, 1979. v. 2, p. 1-116.
13. LAINSON, R. Our present knowledge of the ecology and control of Leishmaniasis in the Amazon region of Brazil. Rev. Soc. bras. Med. trop., 18: 47-56, 1985.
147 LLANOS CUENTAS, A.; ARANA, M.; MONTOYA, Y.; CAMPOS, M. & ROMERO, G. Aislamiento primario de Leishmania sp de pacientes en áreas endémicas del Perú. In: REUNIÃO ANUAL SOBRE PESQUISA BÁSICA EM DOENÇA DE CHAGAS, 11., Caxambu, 1984 Programa e resumos. Ribeirão Preto, PIDE; CNPq, 1984. p. 58.
15. MAYRINK, W.; WILLIAMS, P.; COELHO, M. V.; DIAS, M.; VIANNA MARTINS, A.; MAGALHÃES, P. A.; DA COSTA, C. A.: FALCÃO, A. R.; MELO, M. N. & FALCÃO, A. L. Epidemiology of dermal leishmaniasis in the Rio Doce Valley State of Minas Gerais. Brazil. Ann. trop. Med. Parasit., 73: 124-137, 1979.
16. NETTO, E. M.; CUBA CUBA, C. A.; MARSDEN, P D. & BARRETO, A. C. El método de cultivo "in vitro" como instrumento práctico en el diagnóstico y el aislamiento primario de Leishmania braziliensis braziliensis I. Observaciones de Laboratório. Rev. Soc. bras. Med. trop., 1985 (apresentado para publicación)
17. PETERS, W.: EVANS, D. A. & LANHAM, S. M. The importance of parasite identification in cases of Leishmaniasis. J. roy. Soc. Med., 76: 540-542, 1983.
18. SARVEVA, N. G.; HOLGUIN, A. P.; Mc MAHON PRATT, D. & D'ALESSANDRO, A. Mucocutaneous Leishmaniasis in Colombia: Leishmania braziliensis subspecies diversity. Amer. J. trop. Med. Hyg., 34: 714-720, 1985.
19. SHAW, J. J. & LAINSON, R. The "in vitro" cultivation of members of the Leishmania braziliensis complex. Trans. roy. Soc. irop. Med. Hyg., 75: 127, 1981.
20 URJEL, R.; RECACOECHEA, M,; LA FUENTE, C. & ORELLANA A., H. Simple method for the collection of material from cutaneous and mucocutaneous leishmaniasis lesions. Trans. roy. Soc. trop. Med. Hyg., 77: 882-883, 1973.
21. WALTON, B. C.; SHAW, J. J. & LAINSON, R. Observations on the "in vitro" cultivation of Leishmania braziliensis. J. Parasit., 63: 1118-1119, 1977.
22. WORLD HEALTH ORGANIZATION ù Report of thu Fourth Meeting of the Scientific Working Group on Immunology of Leishmaniasis. UNDP/WORLD/BANK/ WHO TDR^IV; 1982.
23. WIRTH, D. F. & Mc MAHON PRATT, D. Rapid identification of Leishmania species by specific hybridisation of kinetoplast DNA in cutaneous lesions Proc. nat. Acad. Sci. (Wash.), 79: 6999-7003, 1982.

*

Trabajo parcialmente financiado por CNPq (Pro. 403682/82) y OMS/TDR-RE (820526)

Fechas de Publicación

Publicación en esta colección
16 Oct 2012
Fecha del número
Oct 1986

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

[1] 1. AZEVEDO, H. F. & ROITMAN, I. Cultivation of Trypanosoma cruzi in defined media. In: MOREL, C M., ed. Genes and antigens parasites. A laboratory manual. 2nd. ed. Rio de Janeiro, Fundação Oswaldo Cruz, ,1984. p. 29-36.

[2] 2. BARKER, D. C. & BUTCHER, J. The use of PNA probes in the identification of leishmanias: discrimination between isolates of the Leishmania mexicana and L. braziliensis complex. Trans. ray. Soc. trop. med. Hyg., 77: 285-291, 1983.

[3] 3. BARRETO, A. C; CUBA CUBA, C. A.; MARSDEN, P. D.; VEXANAT, J. & BELDER, M. Características epidiemiológicas de Leishmaniose Tegumentar Americana em uma região endêmica do Estado da Bahia. Brasil. 1. Leishmaniose humana. Bol. Ofic. sanit. panamer., 90: 415-424, 1981.

[4] 5. CUBA, C. A.; VEXENAT, A. J.; LLANOS, E. A.; MARSDEN. P. D.; BARRETO. A. C. & ROSA, A. C. Experimental infections of wild caught specimens of Lutzomyia (N) whitmani (Díptera. Psychodidae) and their use for Leishmania identification. In: REUNIÃO ANUAL SOBRE PESQUISA BÁSICA EM DOENÇA DE CHAGAS, 9., Caxambu, 1982. Programa e resumos. Belo Horizonte, CNPq: FINEP, 1982. p. 149.

[5] 6 CUBA CUBA. C. A.; NETTO, M. E.: COSTA, L. J.; ROSA, A. de C ; BARRETO, A. C. & MARSDEN, P. D. Cutaneous leishmaniasis caused by Leishmania braziliensis braziliensis: Improving diagnosis by primary isolation in culture. In: REUNIÃO ANUAL SOBRE PESQUISA BÁSICA EM DOENÇA DE CHAGAS, li.. Caxambu, 1984. Programa e resumos. Ribeirão Preto. PIDE: CNPq. 1984. p. 129.

[6] 7 CUBA CUBA., C. A.; MILES. M. A.; VEXENAT, J. A.; Mc MaHON PRATT. D.: BARKER, D. C: BUTCHER, J. D.; BARRETO, A. C & MARSDEN, P. D. A forus of mucocutaneous leishhmaniasis in Três Braços, Bahia, Brazil. Characterization and identification of Leishmania stocks isolated from man and dogs. Trans. roy. Soc. trop. Med. Hyg., 79: 500-507, 1985.

[7] 8. EVANS, D. A.; LANHAM, S. M.; BALDWIN, C. I. & PETERS, W. Tne isolation and isoenzyme characterization of Leishmania braziliensis sbsp. from patients with cutaneous leishmaniasis acquired in Belize. Trans. roy. Soc. trop. Med. Hyg., 78: 35-42, 1984.

[8] 9. HENDRICKS, L. D. & WRIGHT, N. Diagnosis of cutaneous leishmaniasis by "in vitro" cultivation of saline aspirate in Schneider's Drosophila medium. Amer. J. trop. Med. Hyg., 28: 962-964, 1979.

[9] 10. HENDRICKS, L. D. & CHILDS, G. E. Present knowledge of the in vitro cultivation of Leishmania. In: THE IN VITEO CULTIVATION OF THE PATHOGENS OF TROPICAL DISEASES. Basel, Schwabe & Co. AG, 1980. p. 251-272. (Tropical Diseases Research Series No. 3).

[10] 11. LAINSON, R. & SHAW, J. J. Some problems in studies on parasites of the Leishmania braziliensis complex. In: Ecologie des Leishmanioses. Colloquies Internationaux du CNRS, n.° 239. Montpellier, 1977. p. 83-86.

[11] 12. LAINSON, R. & SHAW, J. J. The role of animals In the epidemiology of South American Leishmaniasis. In: LUMSDEN, W. H. R. & EVANS, D. A., ed. -Biology of the kinetoplastida. London, Academic Press, 1979. v. 2, p. 1-116.

[12] 13. LAINSON, R. Our present knowledge of the ecology and control of Leishmaniasis in the Amazon region of Brazil. Rev. Soc. bras. Med. trop., 18: 47-56, 1985.

[13] 147 LLANOS CUENTAS, A.; ARANA, M.; MONTOYA, Y.; CAMPOS, M. & ROMERO, G. Aislamiento primario de Leishmania sp de pacientes en áreas endémicas del Perú. In: REUNIÃO ANUAL SOBRE PESQUISA BÁSICA EM DOENÇA DE CHAGAS, 11., Caxambu, 1984 Programa e resumos. Ribeirão Preto, PIDE; CNPq, 1984. p. 58.

[14] 15. MAYRINK, W.; WILLIAMS, P.; COELHO, M. V.; DIAS, M.; VIANNA MARTINS, A.; MAGALHÃES, P. A.; DA COSTA, C. A.: FALCÃO, A. R.; MELO, M. N. & FALCÃO, A. L. Epidemiology of dermal leishmaniasis in the Rio Doce Valley State of Minas Gerais. Brazil. Ann. trop. Med. Parasit., 73: 124-137, 1979.

[15] 16. NETTO, E. M.; CUBA CUBA, C. A.; MARSDEN, P D. & BARRETO, A. C. El método de cultivo "in vitro" como instrumento práctico en el diagnóstico y el aislamiento primario de Leishmania braziliensis braziliensis I. Observaciones de Laboratório. Rev. Soc. bras. Med. trop., 1985 (apresentado para publicación)

[16] 17. PETERS, W.: EVANS, D. A. & LANHAM, S. M. The importance of parasite identification in cases of Leishmaniasis. J. roy. Soc. Med., 76: 540-542, 1983.

[17] 18. SARVEVA, N. G.; HOLGUIN, A. P.; Mc MAHON PRATT, D. & D'ALESSANDRO, A. Mucocutaneous Leishmaniasis in Colombia: Leishmania braziliensis subspecies diversity. Amer. J. trop. Med. Hyg., 34: 714-720, 1985.

[18] 19. SHAW, J. J. & LAINSON, R. The "in vitro" cultivation of members of the Leishmania braziliensis complex. Trans. roy. Soc. irop. Med. Hyg., 75: 127, 1981.

[19] 20 URJEL, R.; RECACOECHEA, M,; LA FUENTE, C. & ORELLANA A., H. Simple method for the collection of material from cutaneous and mucocutaneous leishmaniasis lesions. Trans. roy. Soc. trop. Med. Hyg., 77: 882-883, 1973.

[20] 21. WALTON, B. C.; SHAW, J. J. & LAINSON, R. Observations on the "in vitro" cultivation of Leishmania braziliensis. J. Parasit., 63: 1118-1119, 1977.

[21] 22. WORLD HEALTH ORGANIZATION ù Report of thu Fourth Meeting of the Scientific Working Group on Immunology of Leishmaniasis. UNDP/WORLD/BANK/ WHO TDR^IV; 1982.

[22] 23. WIRTH, D. F. & Mc MAHON PRATT, D. Rapid identification of Leishmania species by specific hybridisation of kinetoplast DNA in cutaneous lesions Proc. nat. Acad. Sci. (Wash.), 79: 6999-7003, 1982.