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Parasitology Research

, Volume 106, Issue 5, pp 1249–1252 | Cite as

A new experimental culture medium for cultivation of Leishmania amazonensis: its efficacy for the continuous in vitro growth and differentiation of infective promastigote forms

  • Igor de Almeida Rodrigues
  • Bianca Alcântara da Silva
  • André Luis Souza dos Santos
  • Alane Beatriz Vermelho
  • Celuta Sales Alviano
  • Maria do Socorro Santos RosaEmail author
Short Communication

Abstract

Parasites from the genus Leishmania cause a variety of disease states in humans and other mammals in tropical and subtropical regions, which include cutaneous, mucocutaneous and visceral leishmaniasis. The elaboration of a culture medium for the in vitro cultivation of Leishmania spp., which promotes the growth and differentiation of the parasites, is an important tool for diagnosis, biochemical, biological and immunological studies in the genus. Herein, we have reported the development of a rapid, inexpensive and reliable monophasic culture medium. The novel medium, designated PBHIL, promoted an excellent parasite growth, generating high quantities of promastigotes with long-term viability, and was able to induce cellular differentiation of L. amazonensis promastigotes to the amastigote-like forms (93%). Additionally, we reported the influence of this novel medium on the biochemical characteristics of L. amazonensis and on the interaction of this parasite parasites with mammalian macrophages.

Keywords

Visceral Leishmaniasis Leishmaniasis Brain Heart Infusion Association Index Brain Heart Infusion Medium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This study was supported by grants from Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). The authors also wish thank to Dr. Elvira Saraiva (Departamento de Imunologia, IMPPG-UFRJ) who kindly provided the anti-3A1-La antibody.

References

  1. Alves CR, Corte-Real S, Bourguignon SC, Chaves CS, Saraiva BEM (2005) Leishmania amazonensis: early proteinase activities during promastigote–amastigote differentiation in vitro. Exp Parasitol 109:38–48CrossRefPubMedGoogle Scholar
  2. Balanco JMF, Pral EMF, da Silva S, Bijovsky AT, Mortara RA, Alfieri SC (1998) Axenic cultivation and partial characterization of Leishmania braziliensis amastigote-like stages. Parasitology 116:103–113CrossRefPubMedGoogle Scholar
  3. Bates PA, Robertson CD, Tetley L, Coombs GH (1992) Axenic cultivation and characterization of Leishmania mexicana amastigote like forms. Parasitology 105:193–202CrossRefPubMedGoogle Scholar
  4. Chang KP (1980) Human cutaneous leishmania in a mouse macrophage line: propagation and isolation of intracellular parasites. Science 209:1240–1242CrossRefPubMedGoogle Scholar
  5. Chang KP, Nancy CA, Pearson RD (1986) Intracellular parasitism of macrophages in leishmaniasis: in vitro systems and their applications. Methods Enzymol 132:603–626CrossRefPubMedGoogle Scholar
  6. Chaves CS, Soares DC, Da Silva RP, Saraiva EM (2003) Characterization of the species and stage specificity of two monoclonal antibodies against Leishmania amazonensis. Exp Parasitol 103:152–159CrossRefPubMedGoogle Scholar
  7. Glaser TA (1990) Leishmania major and Leishmania donovani: a method for rapid purification of amastigotes. Exp Parasitol 71:343–345CrossRefPubMedGoogle Scholar
  8. Gupta N, Goyal N, Rastogi AK (2001) In vitro cultivation and characterization of axenic amastigotes of Leishmania. Trends Parasitol 17:150–153CrossRefPubMedGoogle Scholar
  9. Hart DT, Vickerman K, Coombs GH (1981) A quick, simple method for purifying Leishmania mexicana amastigotes in large numbers. Parasitology 82:345–355CrossRefPubMedGoogle Scholar
  10. Ilgoutz SC, McConville MJ (2001) Function and assembly of the Leishmania surface coat. Int J Parasitol 31:899–908CrossRefPubMedGoogle Scholar
  11. Lowry OH, Rozebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  12. Mendonça-Filho RR, Rodrigues IA, Alviano DS, Santos ALS, Soares RM, Alviano CS, Lopes AHCS, Rosa MSS (2004) Leishmanicidal activity of polyphenolic-rich extract from husk fiber of Cocos nucifera Linn. (Palmae). Res Microbiol 155:136–143CrossRefPubMedGoogle Scholar
  13. Muniaraj M, Lal CS, Kumar S, Sinha PK, Das P (2007) Milk of cow (Bos taurus), buffalo (Bubalus bubalis), and goat (Capra hircus): a better alternative than fetal bovine serum in media for primary isolation, in vitro cultivation, and maintenance of Leishmania donovani promastigotes. J Clin Microbiol 45:1353–1356CrossRefPubMedGoogle Scholar
  14. Nicolle C (1908) Culture du parasite du bouton d'Orient. C R Acad Sci 146:842–843Google Scholar
  15. Rosa MSS, Mendonça-Filho RR, Bizzo HR, Rodrigues IA, Soares RMA, Souto-Padrón T, Alviano CS, Lopes AHCS (2003) Antileishmanial activity of linalool-rich essential oil from Croton cajucara Benth. Antimicrob Agents Chemother 47:1895–1901CrossRefGoogle Scholar
  16. Santos ALS, Abreu CM, Alviano CS, Soares RMA (2005) Use of proteolytic enzymes as an additional tool for trypanosomatids identification. Parasitology 130:79–88CrossRefPubMedGoogle Scholar
  17. Schuster FL, Sullivan JJ (2002) Cultivation of clinically significant hemoflagellates. Clin Microbiol Rev 15:374–389CrossRefPubMedGoogle Scholar
  18. Somanna A, Mundodi V, Gedamu L (2002) In vitro cultivation and characterization of Leishmania chagasi amastigote-like forms. Acta Trop 83:37–42CrossRefPubMedGoogle Scholar
  19. Teixeira MCA, Santos RJ, Sampaio RB, Pontes-de-Carvalho L, dos-Santos WLC (2002) A simple and reproducible method to obtain large numbers of axenic amastigotes of different Leishmania species. Parasitol Res 88:963–968CrossRefPubMedGoogle Scholar
  20. Visvesvara GS, Garcia LS (2002) Culture of protozoan parasites. Clin Microbiol Rev 15:327–328CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Igor de Almeida Rodrigues
    • 1
    • 2
  • Bianca Alcântara da Silva
    • 1
  • André Luis Souza dos Santos
    • 1
  • Alane Beatriz Vermelho
    • 1
  • Celuta Sales Alviano
    • 1
  • Maria do Socorro Santos Rosa
    • 1
    Email author
  1. 1.Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS)Universidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  2. 2.Instituto de Química, Centro de TecnologiaUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil

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