FOLIC-ACID THE ESSENTIAL SUPPLEMENT TO BRAIN HEART INFUSION BROTH FORCULTIVATION AND CLONING OF LEISHMANIA-DONOVANI PROMASTIGOTES

Multiplication of virulent and avirulent Leishmania donovani promastig otes was inhibited by brain heart infusion broth (BHI) (3.7%) suppleme nted with haemolysed or whole blood (2.5-10%) or heat-inactivated foet al bovine serum (HIFBS) (10-20%). Peptone, glutamine, tryptophan, puri ne bases, ribonucleosides, deoxyribonucleosides, B-vitamins and vitami n C were evaluated for the reversal of this growth inhibition. Supplem entation of BHI with folic acid further supplemented with haemolysed r abbit blood or HIFBS and haemin supported propagation in serial subpas sages of 2 primary isolates and 3 other virulent strains of L. donovan i and 1 strain of L. major. Folic acid supplemented BHI with 3% rabbit blood in solid phase with 0.7% agar also supported the colony formati on of L. donovani. This manuscript documents the development of high e fficiency media for the cultivation of L. donovani promastigotes in li quid phase and as colonies on agar plates using BHI as a basal medium.