Evaluation of Two Modified Culture Media forLeishmania infantumCultivation Versus Different Culture Media

Abstract

The aim of this study is to improve the cultivation of Leishmania promastigotes without the use of common, semisolid culture media such as Evans' modified Tobie's medium (EMTM), liquid RPMI 1640, and Peptone-yeast extract medium (P-Y). Although EMTM medium permits the growth of a high number of parasites, it is technically difficult to prepare as it requires the use of fresh rabbit blood from animals bred on farms, while RPMI 1640 and P-Y show lower growth rates than the EMTM. There is, therefore, a need to develop new blood-free and time-saving culture systems. The aim of this paper is to propose new modified microbiological media, named RPMI-PY and Tobie-PY, to isolate Leishmania and cultivate parasites for research and diagnostic purposes. This study compares classic culture media to the new media, RPMI-PY and Tobie-PY, and demonstrates that the new media have superior performance in terms of time and parasitic load. The growth rate of the parasite was significantly higher at 24, 48, and 72 hr cultivation, based on counts using Bürker's chambers, when compared to classic media. This study was carried out at the National References Centre for Leishmaniasis (C.Re.Na.L.) where the isolation procedures are conducted daily from a number of different biological matrices.

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

“…Leishmaniasis is a typical example of zoonosis found on all the continents except Australia and Antarctica and is reported by the World Health Organization as being 1 of the major tropical diseases, and 20 million people in the world are infected each year with 400,000 additional phenomena (Castelli et al 2014).…”

In vitro antileishmanial activity of trans-stilbene and terphenyl compounds

“…A strain of Leishmania infantum promastigotes zimodeme 1 (MHON/TN/80/IPT1 MON1) received from the Higher Institute of Health of Rome (Italy) was cultured at 25 C and pH 7.18 in RPMI-PY medium, which consisted of RPMI 1640 (Sigma R0883) supplemented with equal volume of Pepton-yeast medium, 10% fetal bovine serum (FBS), 1% glutamine, 250 mg/mL gentamicin and 500 mg/mL of 5-fluorocytosine (Castelli et al, 2014). To obtain amastigotes cells, promastigotes were cultivated in RPMI-PY, in which the pH was acidified with 1 N HCl to 5.4, and incubated at 37 C.…”

“…In this context, in the last years several natural compounds and their synthetic analogues have been tested against Leishmania, and many of them have demonstrated potential as leishmanicidal agents (Changtam et al, 2010;Tipparaju et al, 2008;Marin et al, 2009;Mishra et al 2009a,b;Hussain et al, 2014;Singh et al, 2014). Stilbenes-based compounds are widely represented in nature (Shen et al, 2009;Riviere et al, 2012) and have become of particular interest to chemists and biologists because of their wide range of biological effects including chemopreventive, antitumor, antioxidant, antimicrobial, anti-inflammatory and antihistaminic activities (Hart, 1981;Fremont, 2000;Young and Chaplin, 2004).…”

Bioactivity of Phycocolloids against the Mediterranean Protozoan Leishmania infantum: An Inceptive Study

“…L. infantum promastigotes (IPT1/MON1, Higher Institute of Health-Rome, Italy) were treated in phosphate buffered saline (PBS) and cultured at 25 • C and pH 7.18 in RPMI-PY medium [19], which consisted of RPMI 1640 (Sigma R0883) supplemented with equal volume of Pepton-yeast medium [20], 10% FBS, 1% glutamine, 250 µg/mL gentamicin, and 500 µg/mL of 5-fluorocytosine. Temperature, differentiation time, and acidification of the medium were used as variables for the preconditioning of the promastigote cultures of L. infatum.…”