SHORT COMMUNICATION 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 Rosa Received: 14 October 2009 / Accepted: 18 January 2010 / Published online: 23 February 2010 # Springer-Verlag 2010 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 differen- tiation 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 character- istics of L. amazonensis and on the interaction of this parasite parasites with mammalian macrophages. Introduction Leishmania spp. are trypanosomatid protozoa that cause a number of important diseases in humans and are endemic in various parts of the world, including Western Asia, the Middle East, Africa, and South America (Ilgoutz and McConville 2001). These organisms are transmitted to the mammalian host by the bite of a sand fly, where they develop in the mid gut as promastigotes. In the mammalian host, the leishmania occupy an intracellular niche as amastigotes, mainly within macrophages of the host. In vitro cultivation of parasitic protozoa that cause human disease is invaluable, as it provides information on the development of parasites that can be used in the contain- ment and eradication of the parasite (Visvesvara and Garcia 2002). The different culture media that have been devel- oped over the past years can be classified into two major categories: semi-solid biphasic and liquid monophasic media. In this sense, the Novy-MacNeal-Nicolle (NNN) medium (Nicolle 1908) and other biphasic-type media are routinely used for initial maintenance of Leishmania promastigote forms (Schuster and Sullivan 2002). How- ever, the collection of blood in the sterile form from rabbits for the NNN medium is a cumbersome process and the maintenance of animal housing is another expensive burden. Due to this, there has been a preference to cultivate promastigotes in liquid media. A readily available medium for parasite culture would be of great use (Muniaraj et al. 2007). The relative ease at which the Leishmania promastigotes can be cultured in vitro has led to the extensive knowledge on the metabolic and molecular machinery of this stage of the parasite (Somanna et al. 2002). In contrast, the difficulty I. A. Rodrigues : B. A. Silva : A. L. S. Santos : A. B. Vermelho : C. S. Alviano : M. S. S. Rosa (*) 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), Bloco I, Ilha do Fundão, Rio de Janeiro, RJ 21941-902, Brazil e-mail: msrrcarvalho@micro.ufrj.br I. A. Rodrigues Instituto de Química, Centro de Tecnologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil Parasitol Res (2010) 106:1249–1252 DOI 10.1007/s00436-010-1775-4