Flow microfluorometric and enzymatic characterization of free-living amoebae

Cytofluorographic analysis of environmental samples has several advantages over the current methods of isolation and classification of free-living amoebae. Using this system, it is possible to rapidly speciate and quantitate mixtures of pathogenic amoebae found in environmental samples. Cytofluorographic analysis of amoebic isolates reduces the time element presently involved in screening environmental sites for the pathogenic amoebae. The cytofluorograf permits speciation of nonthermophilic Naegleria and Acanthamoeba that could not be easily isolated for speciation by conventional methods. Added features of FMF relative to fluorescent microscopy are a high degree of statistical precision, high immunofluorescent titers, elimination of subjectivity, and fading of fluorescence.

Polyacrylamide gel esterase profiles were used in conjunction with FMF to characterize free-living amoebae. Results from the above two systems indicated that two unidentified species of environmentally isolated pathogenic Acanthamoeba were not identical to other known Acanthamoeba characterized in this study. Interspecific-esterase patterns of pathogenic N. Fowleri and nonpathogenic N. Lovaniensis differs. Interspecific esterase profiles from 10 different environmental isolates of each of the Naegleria spp. were identical. These identical esterase patterns seen within the species isolated from different geographical locations suggest that climatic differences do not select for varients of thermophilic Naegleria.