Established laboratory colonies of Lu. longipalpis (from Jacobina, Brazil) and Lu. migonei (from Baturité, Brazil) were used and maintained under standard conditions, as previously described19 (link). A fluorescent strains of L. (V.) braziliensis (XB29 marked with GFP) and L. (L.) infantum (OGVL marked with mCherry) by stable integration of mCherry into the 18 S rRNA locus as previously described20 (link). We have used parasites within less than 10 passages in vitro and were maintained at 23 °C on Medium 199 (Sigma-Aldrich, USA) supplemented with 10% fetal calf serum (Thermo Fisher Scientific, USA), 1% BME vitamins (Sigma-Aldrich), 2% human urine and 250 μg/ml amikin (Bristol-Myers Squibb, USA).
For study of growth curves in-vitro, two initial doses were used: 104 promastigotes/ml and 105 promastigotes/ml. The concentration of parasites was analyzed daily for 7 days, using counting in Burker chamber. The experiments were repeated twice.
For study of growth curves in-vitro, two initial doses were used: 104 promastigotes/ml and 105 promastigotes/ml. The concentration of parasites was analyzed daily for 7 days, using counting in Burker chamber. The experiments were repeated twice.
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