Improved Replication of Autographa calif or nica Nuclear Polyhedrosis Virus in Roller Bottles: Characterization of the Progeny Virus

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Abstract
A reproducible growth curve was established for the propagation of A. californica (Speyer) nuclear polyhedrosis virus (ACMNPV) in a continuous insect line from Spodoptera frugiperda during large-volume production. A newly developed method for quantitation of polyhedra inclusion bodies (PIB) by EM during the growth cycle was compared to hemocytometer counts. The virus particles (VP) to plaque forming units ratio and VP/PIB were established by EM methods. Optimal yields of PIB and cell-free virus with biological activities were obtained in 6 consecutive production lots when the growth curve conditions were followed. The DNA of viruses produced in the 1st and 8th passages in the S. frugiperda cell line was treated with restriction endonucleases and compared with that from the E-2 cloned variant of this virus. The virus was ACMNPV and there were no detectable changes after 8 passages in insect cell culture.

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