Ref ID : 4081
Thomas A. Nerad, Govinda Visvesvara, and Pierre-Marc Daggett; Chemically Defined Media for the Cultivation of Naegleria: Pathogenic and High Temperature Tolerant Species. J.Protozool. 30(2):383-387, 1983
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Chemically defined minimal media for the cultivation of high temperature tolerant and pathogenic Naegleria spp. have been developed. A defined minimal medium, identical for N. fowleri and N. lovaniensis, consists of eleven amino acids (arginine, glycine, histidine, isoleucine, leucine, methionine, phenylalanine, proline, threonine, tryptophan and valine), six vitamins (biotin, folic acid, hemin, pyridoxal, riboflavin, and thiamine), guanosine, glucose, salts and metals. Three of the four strains of Naegleria fowleri tested (ATCC30100, ATCC30863, and ATCC30896) and two strains of N. lovaniensis (ATCC30467 and ATCC30569) could be cultured beyond ten subcultures on this medium. For N. fowleri ATCC30894 diaminopimelic acid, or lysine or glutamic acid was also required. Mean generation time was reduced and population density increased for all strains with the introduction of glutamic acid. Glucose could be eliminated from the minimal medium only if glutamic acid was present. Without glucose, mean generation time increased and population density decreased. Diaminopimelic acid could substitute for lysine for ATCC30894, indicating that Naegleria species may synthesize their lysine via the DAP pathway. Naegleria fowleri ATCC30100 could be adapted to grow without serine or glycine in the minimal medium with glutamic acid added, but with mean generation time increased and population density decreased. The strain could be grown in the minimal medium in the absence of metals. For growth of N. australiensis ATCC30958, modification of the medium by increasing metals ten-fold, substituting guanine for guanosine and adding lysine, glutamic acid, and six vitamins (p-aminobenzoic acid, choline chloride, inositol, vitamin B12, nicotinamide, and Ca pantothenate) was required.