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LB broth-lyophilized Rabbit serum (LLR) as a new and suitable culture medium for cultivation of promastigotes of Leishmania major

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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.

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References

  • Adler S (1964) Leishmania. Adv Parasitol 2:35–96

    Article  CAS  PubMed  Google Scholar 

  • 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

    CAS  PubMed  Google Scholar 

  • Armstrong TC, Patterson JL (1994) Cultivation of Leishmania braziliensis in an economical serum-free medium containing human urine. J Parasitol 80(6):1030–1032

    Article  CAS  PubMed  Google Scholar 

  • Berens RL, Brun R, Krassner SM (1976) A simple monophasic medium for axenic culture of hemoflagellates. J Parasitol 62(3):360–365

    Article  CAS  PubMed  Google Scholar 

  • Bray RS, Rahim GF, Taj-Eldin S (1967) The present state of leishmaniasis in Iraq. Protozoology 2:171–186

    Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Kumar A (2013) Leishmania and Leishmaniasis. Springer, New York

    Book  Google Scholar 

  • 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

    CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  PubMed  PubMed Central  Google Scholar 

  • 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

    Google Scholar 

  • 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

    Article  Google Scholar 

  • 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

    PubMed  PubMed Central  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Taylor AER, Baker JR (1987) In vitro methods for parasite cultivation. Academic Press Ltd, London

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

The authors are thankful to the Director, RVSRI for providing the facilities during the research performing.

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Correspondence to Vahid Nasiri.

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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

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