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A bioassay technique was used to estimate the concentrations of infectious gypsy moth nucleopolyhedrosis virus (NPV) that occur naturaIly in leaf, bark, litter, and soil samples taken from woodland plots in Connecticut and Pennsylvania. These concentrations were then compared to those in samples taken sequentially after treatment of these plots with NPV. Results indicated that NPV is a natural component of the host's habitat, persisting in high concentrations in soil, litter, and on bark for at least one year after natural epizootics. Activity of NPV in spray deposits on foliage and bark was measurable for only 3-15 days after NPV treatment, whereas activity of NPV liberated from larval cadavers onto bark was measurable in high concentrations through May of the year foIlowing treatment. The application of NPV at the rate of 2.5 x 10 12 polyhedral inclusion bodies (PIB)/ha to an area already containing high concentrations of naturally occurring NPV did not cause an increase in the environmental NPV load. Although the application of NPV at the rate of 5 x 10 12 PIB/ha to plots containing low natural concentrations of NPV resulted in measurable increases in the NPV load in these plots, these increases were not, overaIl' significantly higher than those occurring in a control plot that was not treated with NPV.

The adaptation of pathogens to either their hosts or to environmental conditions is the focus of many current ecological studies. In this work we compared the ability of six spatially-distant Lymantria dispar (gypsy moth) multiple nucleopolyhedrovirus (LdMNPV) strains (three from eastern North America and three from central Asia) to induce acute infection in gypsy moth larvae. We also sequenced the complete genome of one Asian (LdMNPV-27/0) and one North American (LdMNPV-45/0) strain which were used for bioassay. We found that all of the North American virus strains, with the exception of one, demonstrated higher potency than the Asian virus strains, either in North American (Lymantria dispar) larvae or, in Asian (Lymantria dispar asiatica) larvae. Complete genome sequencing revealed two gene deletions in the LdMNPV-27/0 strain: the virus enhancin factor gene (vef-1) and the baculovirus repeated orf gene (bro-p). These deletions were not seen in the LdMNPV-45/0 strain nor in other American strains available in archiving systems. We also found deletions of the bro-e and bro-o genes in LdMNPV-45/0 strain but not in the LdMNPV-27/0 strain. The phylogenetic inference with an alignment of the 37 core gene nucleotide sequences revealed the close relationship of the LdMNPV-45/0 strain with other American strains accessed in GenBank (Ab-a624 and 5-6) while the LdMNPV-27/0 strain was clustered together with the LdMNPV-3054 strain (isolated in Spain) instead of predicted clustering with LdMNPV- 3029 (isolated in Asia). Our study demonstrated that first, different LdMNPV isolates from the same metapopulations of L. dispar exhibit little or no difference in the degree of virulence towards host larvae and second, that locality of host population is not an important driver of LdMNPV virulence. Virulence of LdMNPV is determined only by viral genetics. The genetic differences between North American and Central Asian virus strains are discussed.