It has been confirmed that two species of free-living amoebae of the genus Naegleria, i.e., N. fowleri and N. australiensis are intective to man and cause fatal meningoencephalitis known as primary amoebic meningoencephalitis. About 130 cases have been reported worldwide with two cases in China: one in Beijing caused by Naegleria fowleri, and the other in Shanghai with unknown causative agent. From a survey of pathogenic free-living amoebae in Shanghai, we found a strain WO42302 isolated from the cooling water of a power plant in Shanghai in April 1986. It could be fatal to the mouse. Culture of the brain tissue of the dead mice was positive for the amoeba. According to the morphology of trophozoites and their ability to transform into a temporary flagellate stage, the parasite belongs to the genus Naegleria. But the strain WO42302 was different from N. fowleri, by several ultrastructural characteristics, the protein and isoenzyme patterns of acid phosphatase, alkaline phosphatase and lactate dehydrogenase and its lower virulence. The results obtained in the present study infer that the pathogenic strain WO42302 may belong to the species N. australiensis, which is new to China.