Comparison of Three Different Techniques for Follow-up of Free-living Amebae after Drug Therapy

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Chutatip Siripanth
Benjanee Punpoowong



The experiment compared fresh observation, staining, and tissue culture methods for detectionof pathogenic free-living amebae after drug therapy. Fresh observations, using a compoundmicroscope and phase-contrast optics were performed. Giemsa, Gram-chromotrope, and Thomasstains were used. The ameba trophozoites were inoculated into Hep-2 and primary chicken kidneycells and their survival was determined within 24 hours. Among the three methods, tissue culturecould detect the ameba trophozoites from 24 hours’ post-inoculation. Although atypical characterswere demonstrated by fresh observation, and the survival of ameba trophozoites could not bedetermined by staining methods, tissue culture demonstrated the viability of ameba trophozoites.Therefore, tissue culture was the best technique for detecting the survival and viability of pathogenicfree-living amebae after drug therapy.

Keywords: Naegleria, Acanthamoeba, phase-contrast, Giemsa stain, Gram-chromotrope stain, Thomasstain, tissue culture

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