Acanthamoeba sp. from the Philippines: electron microscopy studies on naturally occurring bacterial symbionts

The isolation of two plasmid-like circular DNAs, measuring 52 and 42 kbp, from an Acanthamoeba sp. from the Philippines has led to the demonstration of a bacterial endosymbiont occurring in this free-living amoeba. The 52-kbp band hybridized with a short sequence of cytochrome b gene and was identified as the mitochondrial DNA, whereas the 42-kbp band was identified as plasmid DNA of the bacterial symbionts on the basis of electron microscopy. The endosymbionts are gram-negative, rod-shaped bacteria measuring approximately 1.3 x 0.43 microns and numbering about eight to ten cells per section. They are randomly distributed in both cysts and trophozoites and are surrounded neither by a phagolysosomal membrane nor by a clear or electron-translucent region. The endosymbiont membrane appears to have a close association with ribosomes, which are seen to be more concentrated within the vicinity of the symbionts than elsewhere within the cytoplasm. Attempts to grow the symbionts and the amoebae separately have failed.