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Leishmania donovani development in Phlebotomus argentipes: comparison of promastigote- and amastigote-initiated infections



Leishmania parasites alternate in their life cycle between promastigote stages that develop in the gut of phlebotomine sand flies and amastigotes residing inside phagocytic cells of vertebrate hosts. For experimental infections of sand flies, promastigotes are frequently used as this way of infection is technically easier although ingestion of promastigotes by sand flies is unnatural. Here we aimed to answer a critical question, to what extent do promastigote-initiated experimental infections differ from those initiated with intracellular amastigotes. We performed side-by-side comparison of Leishmania development in Phlebotomus argentipes females infected alternatively with promastigotes from log-phase cultures or amastigotes grown ex vivo in macrophages. Early stage infections showed substantial differences in parasite load and representation of morphological forms. The differences disappeared along the maturation of infections; both groups developed heavy late-stage infections with colonization of the stomodeal valve, uniform representation of infective metacyclics and equal efficiency of transmission. The results showed that studies focusing on early phase of Leishmania development in sand flies should be initiated with intracellular amastigotes. However, the use of promastigote stages for sand fly infections does not alter significantly the final outcome of Leishmania donovani development in P. argentipes and their transmissibility to the vertebrate host.

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This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (, which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited.

Corresponding author

*Corresponding author: Department of Parasitology, Faculty of Science, Charles University, Vinicna 7, 128 44 Prague 2, Czech Republic. E-mail:


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Anjili, C., Langat, B., Lugalia, R., Mwanyumba, P., Ngumbi, P., Mbati, P. A., Githure, J. and Tonui, W. K. (2006). Estimation of the minimum number of Leishmania major amastigotes required for infecting Phlebotomus duboscqi (Diptera: Psychodidae). East African Medical Journal 83, 6871.
Bates, P. A. (1993). Axenic culture of Leishmania amastigotes. Parasitology Today 9, 143146.
Bates, P. A. (2007). Transmission of Leishmania metacyclic promastigotes by phlebotomine sand flies. International Journal for Parasitology 37, 10971106.
Borovsky, D. and Schlein, Y. (1987). Trypsin and chymotrypsin-like enzymes of the sandfly Phlebotomus papatasi infected with Leishmania and their possible role in vector competence. Medical and Veterinary Entomology 1, 235242.
Chang, K. P. (1980). Human cutaneous leishmania in a mouse macrophage line: propagation and isolation of intracellular parasites. Science 209, 12401242.
Charest, H. and Matlashewski, G. (1994). Developmental gene expression in Leishmania donovani: differential cloning and analysis of an amastigote-stage-specific gene. Molecular and Cellular Biology 14, 29752984.
Dostalova, A. and Volf, P. (2012). Leishmania development in sand flies: parasite–vector interactions overview. Parasites & Vectors 5, 276.
Freitas, V. C., Parreiras, K. P., Duarte, A. P., Secundino, N. F. and Pimenta, P. F. (2012). Development of Leishmania (Leishmania) infantum chagasi in its natural sandfly vector Lutzomyia longipalpis . The American Journal of Tropical Medicine and Hygiene 86, 606612.
Gupta, N., Goyal, N. and Rastogi, A. K. (2001). In vitro cultivation and characterization of axenic amastigotes of Leishmania . Trends in Parasitology 17, 150153.
Holzer, T. R., McMaster, W. R. and Forney, J. D. (2006). Expression profiling by whole-genome interspecies microarray hybridization reveals differential gene expression in procyclic promastigotes, lesion-derived amastigotes, and axenic amastigotes in Leishmania mexicana . Molecular and Biochemical Parasitology 146, 198218.
Kimblin, N., Peters, N., Debrabant, A., Secundino, N., Egen, J., Lawyer, P., Fay, M. P., Kamhawi, S. and Sacks, D. (2008). Quantification of the infectious dose of Leishmania major transmitted to the skin by single sand flies. Proceedings of the National Academy of Sciences of the United States of America 105, 1012510130.
Lehane, M. J. (1997). Peritrophic matrix structure and function. Annual Review of Entomology 42, 525550.
Maia, C., Seblova, V., Sadlova, J., Votypka, J. and Volf, P. (2011). Experimental transmission of Leishmania infantum by two major vectors: a comparison between a viscerotropic and a dermotropic strain. PLoS Neglected Tropical Disieses 5, e1181.
Mary, C., Faraut, F., Lascombe, L. and Dumon, H. (2004). Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. Journal of Clinical Microbiology 42, 52495255.
McCall, L. I. and Matlashewski, G. (2012). Involvement of the Leishmania donovani virulence factor A2 in protection against heat and oxidative stress. Experimental Parasitology 132, 109115.
Myskova, J., Votypka, J. and Volf, P. (2008). Leishmania in sand flies: comparison of quantitative polymerase chain reaction with other techniques to determine the intensity of infection. Journal of Medical Entomology 45, 133138.
Pescher, P., Blisnick, T., Bastin, P. and Späth, G. F. (2011). Quantitative proteome profiling informs on phenotypic traits that adapt Leishmania donovani for axenic and intracellular proliferation. Cellular Microbiology 13, 978991.
Pimenta, P. F., Modi, G. B., Pereira, S. T., Shahabuddin, M. and Sacks, D. L. (1997). A novel role for the peritrophic matrix in protecting Leishmania from the hydrolytic activities of the sand fly midgut. Parasitology 115, 359369.
Pruzinova, K., Sadlova, J., Seblova, V., Homola, M., Votypka, J. and Volf, P. (2015). Comparison of bloodmeal digestion and the peritrophic matrix in four sand fly species differing in susceptibility to Leishmania donovani . PLoS ONE 10, e0128203.
Ramalho-Ortigão, J. M. and Traub-Csekö, Y. M. (2003). Molecular characterization of Llchit1, a midgut chitinase cDNA from the leishmaniasis vector Lutzomyia longipalpis . Insect Biochemistry and Molecular Biology 33, 279287.
Ramalho-Ortigão, J. M., Kamhawi, S., Joshi, M. B., Reynoso, D., Lawyer, P. G., Dwyer, D. M., Sacks, D. L. and Valenzuela, J. G. (2005). Characterization of a blood activated chitinolytic system in the midgut of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi . Insect Molecular Biology 14, 703712.
Rochette, A., Raymond, F., Corbeil, J., Ouellette, M. and Papadopoulou, B. (2009). Whole-genome comparative RNA expression profiling of axenic and intracellular amastigote forms of Leishmania infantum . Molecular and Biochemical Parasitology 165, 3247.
Rogers, M. E., Chance, M. L. and Bates, P. A. (2002). The role of promastigote secretory gel in the origin and transmission of the infective stage of Leishmania mexicana by the sandfly Lutzomyia longipalpis . Parasitology 124, 495507.
Rogers, M. E., Hajmova, M., Joshi, M. B., Sadlova, J., Dwyer, D. M., Volf, P. and Bates, P. A. (2008). Leishmania chitinase facilitates colonization of sand fly vectors and enhances transmission to mice. Cellular Microbiology 10, 13631372.
Sacks, D. L. and Perkins, P. V. (1985). Development of infective stage Leishmania promastigotes within phlebotomine sand flies. The American Journal of Tropical Medicine and Hygiene 34, 456459.
Schlein, Y. and Jacobson, R. L. (1998). Resistance of Phlebotomus papatasi to infection with Leishmania donovani is modulated by components of the infective bloodmeal. Parasitology 117, 467473.
Seblova, V., Volfova, V., Dvorak, V., Pruzinova, K., Votypka, J., Kassahun, A., Gebre-Michael, T., Hailu, A., Warburg, A. and Volf, P. (2013). Phlebotomus orientalis sand flies from two geographically distant Ethiopian localities: biology, genetic analyses and susceptibility to Leishmania donovani . PLoS Neglected Tropical Diseases 7, e2187.
Stamper, L. W., Patrick, R. L., Fay, M. P., Lawyer, P. G., Elnaiem, D. E., Secundino, N., Debrabant, A., Sacks, D. L. and Peters, N. C. (2011). Infection parameters in the sand fly vector that predict transmission of Leishmania major . PLoS Neglected Tropical Disieses 5, e1288.
Stierhof, Y. D., Bates, P. A., Jacobson, R. L., Rogers, M. E., Schlein, Y., Handman, E. and Ilg, T. (1999). Filamentous proteophosphoglycan secreted by Leishmania promastigotes forms gel-like three-dimensional networks that obstruct the digestive tract of infected sandfly vectors. European Journal of Cell Biology 78, 675689.
Sadlova, J. and Volf, P. (2009). Peritrophic matrix of Phlebotomus duboscqi and its kinetics during Leishmania major development. Cell and Tissue Research 337, 313325.
Sadlova, J., Price, H. P., Smith, B. A., Votypka, J., Volf, P. and Smith, D. F. (2010). The stage-regulated HASPB and SHERP proteins are essential for differentiation of the protozoan parasite Leishmania major in its sand fly vector, Phlebotomus papatasi . Cellular Microbiology 12, 17651779.
Sadlova, J., Yeo, M., Seblova, V., Lewis, M. D., Mauricio, I., Volf, P. and Miles, M. A. (2011). Visualisation of Leishmania donovani fluorescent hybrids during early stage development in the sand fly vector. PLoS ONE 6, e19851.
Volf, P. and Volfova, V. (2011). Establishment and maintenance of sand fly colonies. Journal of Vector Ecology 36, S1S9.
Walters, L. L. (1993). Leishmania differentiation in natural and unnatural sand fly hosts. Journal of Eukaryotic Microbiology 40, 196206.
Warburg, A. and Schlein, Y. (1986). The effect of post-bloodmeal nutrition of Phlebotomus papatasi on the transmission of Leishmania major . The American Journal of Tropical Medicine and Hygiene 35, 926930.



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