Species of the family Mimiviridae habor a giant virus called Mimivirus. The repeated isolation of the giant virus “KSL5” from bark of a plane tree as its original site is reported. For this purpose, the original parasite-free amoebal host Saccamoeba sp. was co-cultivated with crushed and filtered samples from bark of various plane trees, from ground or litoral sediment of a creek, respectively. Only after association with the crushed bark of that plane tree from which it was isolated previously, Mimivirus-like organisms could be detected within few trophozoites of this free-living amoeba. Electron microscopy revealed that the virus factories developing within the cytoplasm of the Saccamoeba host produced solely a great number of polygonal capsids, whereas no production of tiny virophages could be observed as seen and described during the first isolation.