ABSTRACT

Objective:

Human African trypanosomiasis, also known as sleeping sickness, is a parasitic disease in which Glossina is transmitted by human intervention and Trypanosoma b. rhodosiense and Trypanosoma b. gambiense are the causative agents Production of parasites in axenic cultures provides great advantage in parasite biochemistry, immunological, physiological and molecular studies. In this study, it is aimed to determine the medium which will produce in vigorous amount of Trypanosoma b. rhodosiense and Trypanasoma cruzi and to establish a new medium.

Methods:

In this study, Trypanosoma b. rhodosiense and Trypanasoma cruzi strains stored in Manisa Celal Bayar University Parasite Bank will be removed from liquid nitrogen tank under suitable conditions, planted in Medium I, Medium II, Medium III and newly developed medium. Reproductive densities of the media will be statistically analyzed on Thoma lamina depending on the time, using the Sidak’s multiplequality test.

Results:

As a result of this study, it has been concluded that the best medium, to produce abundantly Trypanosoma b. rhodosiense and Trypanasoma cruzi strains, to be used in diagnosis and active substance screenings, molecular studies, metabolic analyzes and drug studies is the medium IV.

Conclusion:

This study is one of the first studies related to the production of Trypanosoma species in Turkey and planned to provide a basis for the studies of African sleeping disease, Chagas disease and their agents.

Keywords: Trypanosoma b. rhodosiense, Trypanosoma cruzi, medium, trypanosomiasis, Turkey

References

Trypanosomiasis-human-african-(sleeping-sickness). www.who.int [Internet]. (cited 2019 February 5). Available from: URL: https://www.who.int/news-room/fact-sheets/detail/trypanosomiasis-human-african-(sleeping-sickness)

Franco JR, Simarro PP, Diarra A, Jannin JG. Epidemiology of human African trypanosomiasis. Clin Epidemiol 2014;6:257-75.

Rassi A, Rassi A, Marin-Neto JA. Chagas disease. Lancet 2010;375(9723):1388-402.

Novy FG, McNeal WJ. On the Cultivation of Trypanosoma brucei. J Infect Dis 1904;1:1-30.

Weinman D. Cultivation of African sleeping sickness trypanosomes on improved, simple, cell-free medium. Proc Soc Exp Biol Med 1946;63:456-8.

Razgha A. Über die Züchtung der menschenpathogenen Trypanosomen. Zeitschrift fiir Parasitenkd 1929;2:55-66. Available from: URL: https://link.springer.com/content/pdf/10.1007/BF02120353.pdf

Reichenow E. Die Züchtung der patogenen Trypanosomen. Arch Schiffsu Tropenhyg 1934;38:292-302.

Brutsaert P, Henrard C. L’hemoculture comme moyen auxiliaire de diagnostic de la maladie du sommeil. CR Soc Biol 1938;127:1469-72.

Chang SL. Studies on Hemoflagellates. IV. Observations Concerning some Biochemical Activities in Culture and espiration of Three Species of Leishmania and Trypanosoma cruzi. J Infect Dis 1948;82:109-18.

Hendricks LD, Wood DE, Hajduk ME. Haemoflagellates: commercially available liquid media for rapid cultivation. Parasitology 1978;76:309-16.

Childs GE, McRoberts MJ, Moussa MA. Systems for the in vitro large-scale propagation of New World Leishmania. Ann Trop Med Parasitol 1979;73:395-6.

Chaudhuri G, Ghoshal K, Pal S, Sen S, Banerjee AB. A new medium for large scale production of Leishmania donovani promastigotes for biochemical studies. Indian J Med Res 1986;84:457-60.

O’Daly JA, Rodriguez MB. Differential growth requirements of several Leishmania spp. in chemically defined culture media. Acta Trop 1988;45:109-26.

Kar K, Mukerji K, Naskar K, Bhattacharya A, Ghosh DK. Leishmania donovani: a chemically defined medium suitable for cultivation and cloning of promastigotes and transformation of amastigotes to to promastigotes. J Protozool 1990;37:277-9.

Gibson WC. Will the real Trypanosoma b. gambiense please stand up. Parasitol Today 1986;2:255-7.

Hirumi H, Doyle JJ, Hirumi K. African trypanosomes: cultivation of animal-infective Trypanosoma brucei in vitro. Science 1977;196(4293):992-4.

Duszenko M, Ferguson MA, Lamont GS, Rifkin MR, Cross GA. Cysteine eliminates the feeder cell requirement for cultivation of Trypanosoma brucei bloodstream forms in vitro. J Exp Med 1985;162:1256-63.

Hirumi H, Hirumi K. Continuous cultivation of Trypanosoma brucei blood stream forms in a medium containing a low concentration of serum protein without feeder cell layers. J Parasitol 1989;75:985-9.

Vassella E, Boshart M. High molecular mass agarose matrix supports growth of bloodstream forms of pleomorphic Trypanosoma brucei strains in axenic culture. Mol Biochem Parasitol 1996;82:91-105.

Vassella E, Krämer R, Turner CM, Wankell M, Modes C, van den Bogaard M, et al. Deletion of a novel protein kinase with PX and FYVE-related domains increases the rate of differentiation of Trypanosoma brucei. Mol Microbiol 2001;41:33-46.

McCulloch R, Vassella E, Burton P, Boshart M, Barry JD. Transformation of Monomorphic and Pleomorphic Trypanosoma brucei. Methods Mol Biol 2004;262:53-86.

Van Reet N, Pyana PP, Deborggraeve S, Büscher P, Claes F. Trypanosoma brucei gambiense: HMI-9 medium containing methylcellulose and human serum supports the continuous axenic in vitro propagation of the bloodstream form. Exp Parasitol 2011;128:285-90.

Luz ZM, Coutinho MG, Cançado JR, Krettli AU. [Hemoculture: sensitive technique in the detection of Trypanosoma cruzi in chagasic patients in the chronic phase of Chagas disease]. Rev Soc Bras Med Trop 1994;27:143-8.

D’Alessandro-Bacigalupo A, Saravia NG. Trypanosoma rangeli. Parasit Protozoa 1992;2;1-54.

Saldaña A, Sousa OE. Trypanosoma rangeli: epimastigote immunogenicity and cross-reaction with Trypanosoma cruzi. J Parasitol 1996;82:363-6.

Howard MK, Sayers G, Miles MA. Leishmania donovani metacyclic promastigotes: transformation in vitro, lectin agglutination, complement resistance, and infectivity. Exp Parasitol 1987;64:147-56.

Ferreira KA, Lemos-Júnior PE, Lages-Silva E, Ramírez LE, Pedrosa AL. Human urine stimulates in vitro growth of Trypanosoma cruzi and Trypanosoma rangeli. Parasitol Res 2007;101:1383-8.

The Production of Trypanosoma Brucei Rhodesiense, Cause of African Sleeping Sickness, and Trypanosoma Cruzi, Cause of American Chagas Disease, on Different Medias and Testing a New Media

ABSTRACT

References