Abstract
Fetal calf serum is the major part and the most expensive ingredient of the Leishmania culture media. Here, the efficacy of the LB broth-lyophilized Rabbit serum medium (LLR) was evaluated in cultivation of Leishmania major. Conventional Luria–Bertani (LB) broth medium was prepared and autoclaved for 15 min at 121 °C and then lyophilized Rabbit serum was added at the 1, 2.5, 5 and 10 % final concentrations. The efficacy of medium was evaluated by assessing the growth ability and replication pattern of the promastigotes of L. major. According to our finding, the LLR medium with 5–10 % lyophilized Rabbit serum supported the growth of the parasites and can be used for cultivation of Leishmanian parasites with acceptable In vivo infectivity for research purpose. The ability of the parasites to survive and proliferating in the presence of lyophilized Rabbit serum indicating that this serum is a good nutritional source. This study opens a new way to make low-cost medium that could be used in cultivation of Leishmanian parasites.
Similar content being viewed by others
References
Adler S (1964) Leishmania. Adv Parasitol 2:35–96
Ali SA, Iqbal J, Ahmad B, Masoom M (1998) A semisynthetic fetal calf serum-free liquid medium for in vitro cultivation of Leishmania promastigotes. Am J Trop Med Hyg 59:163–165
Armstrong TC, Patterson JL (1994) Cultivation of Leishmania braziliensis in an economical serum-free medium containing human urine. J Parasitol 80(6):1030–1032
Berens RL, Brun R, Krassner SM (1976) A simple monophasic medium for axenic culture of hemoflagellates. J Parasitol 62(3):360–365
Bray RS, Rahim GF, Taj-Eldin S (1967) The present state of leishmaniasis in Iraq. Protozoology 2:171–186
Castelli G, Galante A, Lo Verde V et al (2014) Evaluation of two modified culture media for Leishmania infantum cultivation versus different culture media. J Parasitol 100:228–230
García N, Moreno I, Alvarez J et al (2014) Evidence of Leishmania infantum infection in rabbits (Oryctolagus cuniculus) in a natural area in Madrid, Spain
Ghoshal K, Sen S, Pal S, Banerjee AB (1986) Nutrition of Leishmania donovani donovani: growth in new semidefined & completely chemically defined media. Indian J Med Res 84:461
Herman R (1966) Studies of the Numbers and Morphology of the Intracellular Form of Leishmania donovani grown in cell culture. J Protozool 13:408–418
Kumar A (2013) Leishmania and Leishmaniasis. Springer, New York
Merlen T, Sereno D, Brajon N et al (1999) Leishmania spp.: completely defined medium without serum and macromolecules (CDM/LP) for the continuous in vitro cultivation of infective promastigote forms. Am J Trop Med Hyg 60:41–50
Molina R, Jiménez MI, Cruz I et al (2012) The hare (Lepus granatensis) as potential sylvatic reservoir of Leishmania infantum in Spain. Vet Parasitol 190:268–271
Moreno I, Álvarez J, García N et al (2014) Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test. Vet Parasitol 199:264–267. doi:10.1016/j.vetpar.2013.10.010
Nasiri V (2013) Sheep blood-LB agar base medium (SLM) as a simple and suitable medium for the cultivation of Leishmania major promastigotes. Parasitol Res 112:3741–3742. doi:10.1007/s00436-013-3560-7
Nasiri V, Dalimi A, Habibi GH et al (2011) Use of chicken serum as a good replacement for the fetal calf serum in cultivation of promastigotes of Leishmania major. Arch Razi 66:59–64
Nasiri V, Dalimi A, Ghaffarifar F (2013a) Use of chicken (Gallus gallus) serum as a costly replacement for the fetal calf serum in cultivation of promastigotes of Leishmania infantum. Asian Pacific J Trop Dis 3:169–173. doi:10.1016/S2222-1808(13)60034-1
Nasiri V, Karimi G, Dalimi A et al (2013b) Effects of sheep and mouse urine on the growth pattern of Leishmania major promastigotes. Biomed Res Int. doi:10.1155/2013/748592
Newman C (2003) Serum-free cell culture—the ethical, scientific focus on alternatives is an organisation working towards alternatives, pp 941–942
Pal JK, Joshi-Purandare M (2001) Dose-dependent differential effect of hemin on protein synthesis and cell proliferation in Leishmania donovani promastigotes cultured in vitro. J Biosci 26:225–231
Ruiz-Fons F, Ferroglio E, Gortázar C (2013) Leishmania infantum in free-ranging hares, Spain, 2004–2010. Euro Surveill 18:20541. doi:10.2807/1560-7917.ES2013.18.30.20541
Schuster FL, Schuster FL, Sullivan JJ, Sullivan JJ (2002) Cultivation of clinically significant hemoflagellates. Society 15:374–389. doi:10.1128/CMR.15.3.374
Shamsuzzaman SM, Furuya M, Korenaga M et al (1999) Use of urine samples from healthy humans, nephritis patients or other animals as an alternative to foetal calf serum in the culture of Leishmania (L.) donovani in vitro. Ann Trop Med Parasitol 93:613–620
Taylor AER, Baker JR (1987) In vitro methods for parasite cultivation. Academic Press Ltd, London
The World Health Organization (2004) The World Health Report 2004: changing history, annex table 3: burden of disease in DALYs by cause, sex, and mortality stratum in WHO regions, estimates for 2002
The World Health Organization (2010) Control of the leishmaniases: report of a meeting of the WHO Expert Commitee on the Control of Leishmaniases, Geneva, 22–26 March 2010. World Health Organization, Geneva
Trager W (1953) The development of Leishmania donovani in vitro at 37 °C effects of the kind of serum. J Exp Med 97:177–188
Acknowledgments
The authors are thankful to the Director, RVSRI for providing the facilities during the research performing.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
We declare that we have no conflict of interest.
Rights and permissions
About this article
Cite this article
Nasiri, V., Dalimi, A. & Ghaffarifar, F. LB broth-lyophilized Rabbit serum (LLR) as a new and suitable culture medium for cultivation of promastigotes of Leishmania major . J Parasit Dis 41, 247–251 (2017). https://doi.org/10.1007/s12639-016-0786-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12639-016-0786-1