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Abstract
Three isolates of Entomophaga maimaiga, a fungal pathogen of gypsy moth (Lymantria dispar), were stored immersed in liquid nitrogen (-196 C) as naturally occurring protoplasts for approximately 7 years. All isolates survived although with diminished ability to produce conidia and azygospores. Days from infection to host death increased for all isolates after the storage period. Soon after isolation, two of the isolates differed in levels of azygospores produced at 25 C, while after storage, differences among isolates persisted but had changed; at 25 C, all isolates differed from each other in levels of conidial production, and one isolate no longer produced azygospores. Days from infection until host death for one isolate were significantly longer in comparison with other isolates both before and after storage, and this latter isolate also grew slowly in vitro as well as in vivo. Our results suggest that further investigations are warranted to study potential changes in characteristics of pathogenicity during maintenance of fastidious fungal entomopathogens under liquid nitrogen in a culture collection.