Studies on the cloning of few-coiled type of spore in entomopathogenic microsporidia

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The purpose of this study was to clone entomogenous microsporidia by infecting an insect cultured cell with a single spore. In an attempt to isolate the octospore of Vairimorha necatrix that had been propagated in silkworm larvea reared at 16゜C. In the genus Nusema, one of the two types of the spore, the few-coiled type was recovered from cultured Helicoverpa zea cells infected with Nosema furnacalis and cultured Antheraea eucalypti cells infected with N. bombycis.A single spore was recovered from each suspension of few-coiled type spores and transferred to a cell culture using a micromanupulator.Although the developmental stages of N. furnacalis were detected in cultured H. zea cells after 30-day incubation, those of N. bombycis were not. When a single spore of many-coiled type of N. bombycis spore was transferred to cell cultures after alkaline activation, various developmental stages were detected in cultured A. eucalypti cells after 30-day incubation. The cloned microsporidia from a single spore could be subcultured by supplementing the culture with the healthy cells.