Application of Balsam Fir Sawfly Nucleopolyhedrovirus against its Natural Host Neodiprion abietis (Hymenoptera : Diprionidae)

University of New Brunswick (Canada)Natural Resources Canada, Canadian Forest Service -Atlantic Forestry CentreProceedings : IUFRO Kanazawa 2003 "Forest Insect Population Dynamics and Host Influences"., Scedule:14－19 September 2003, Vemue: Kanazawa Citymonde Hotel, Kanazawa, Japan, Joint metting of IUFRO working groups : 7.01.02 Tree resistance to Insects | 7.03.06 Integrated management of forset defoloating insects | 7.03.07 Population dynamics of forest insects, Sponsored by: IUFRO-J | Ishikawa Prefecture | Kanazawa City | 21st-COE Program of Kanazawa University, Editors: Kamata, Naoto | Liebhold, Nadrew M. | Quiring, Dan T. | Clancy, Karen M

Complete Sequence, Analysis and Organization of the Orgyia leucostigma Nucleopolyhedrovirus Genome

The complete genome of the Orgyia leucostigma nucleopolyhedrovirus (OrleNPV) isolated from the whitemarked tussock moth (Orgyia leucostigma, Lymantridae: Lepidoptera) was sequenced, analyzed, and compared to other baculovirus genomes. The size of the OrleNPV genome was 156,179 base pairs (bp) and had a G+C content of 39%. The genome encoded 135 putative open reading frames (ORFs), which occupied 79% of the entire genome sequence. Three inhibitor of apoptosis (ORFs 16, 43 and 63), and five baculovirus repeated ORFs (bro-a through bro-e) were interspersed in the OrleNPV genome. In addition to six direct repeat (drs), a common feature shared among most baculoviruses, OrleNPV genome contained three homologous regions (hrs) that are located in the latter half of the genome. The presence of an F-protein homologue and the results from phylogenetic analyses placed OrleNPV in the genus Alphabaculovirus, group II. Overall, OrleNPV appears to be most closely related to group II alphabaculoviruses Ectropis obliqua (EcobNPV), Apocheima cinerarium (ApciNPV), Euproctis pseudoconspersa (EupsNPV), and Clanis bilineata (ClbiNPV)

Transmission of a Gammabaculovirus within Cohorts of Balsam Fir Sawfly (Neodiprion abietis) Larvae

Nucleopolyhedroviruses (NPV: Gammabaculovirus: Baculoviridae) of diprionid sawflies (Diprionidae: Hymenoptera) are highly host specific and only infect the midgut epithelium. While still alive, infected sawfly larvae excrete NPV-laden diarrhea that contaminates food sources. The diarrhea can then be consumed by conspecific larvae, resulting in rapid horizontal transmission of the virus. To better understand the efficacy of Gammabaculovirus-based biological control products, the horizontal spread of such a virus (NeabNPV) within cohorts of balsam fir sawfly (Neodiprion abietis) larvae was studied by introducing NeabNPV-treated larvae into single-cohort groups at densities similar to those observed during the increasing (field study) and peak (laboratory study) phases of an outbreak. In field studies (~200 N. abietis larvae/m2 of balsam fir (Abies balsamea) foliage), NeabNPV-induced mortality increased positively in a density-dependent manner, from 23% (in control groups) to 51% with the addition of one first-instar NeabNPV-treated larva, to 84% with 10 first–instar-treated larvae. Mortality was 60% and 63% when one or 10 NeabNPV-treated third-instar larva(e), respectively, were introduced into groups. Slightly higher levels of NeabNPV-induced mortality occurring when NeabNPV-treated larvae were introduced into first- rather than third-instar cohorts suggests that early instars are more susceptible to the virus. In the laboratory (~1330 N. abietis larvae/ m2 of foliage), NeabNPV-caused mortality increased from 20% in control groups to over 80% with the introduction of one, five or 10 NeabNPV-treated larvae into treatment groups of first-instar larvae

Histology of the Larval Neodiprion abietis (Hymenoptera: Diprionidae) Digestive Tract

The alimentary canal of Neodiprion abietis larvae is a straight tube divided into foregut, midgut, and hindgut. Posterior to the mouth, the foregut is further divided into the pharynx, esophagus (crop), and proventriculus, all of which are lined with cuticle. A pair of muscular, chitin-lined pouches branch off the anterior foregut and lie lateral to the alimentary canal. Gastric caeca are located at the anterior end of the midgut, where the peritrophic membrane is formed and was observed throughout the midgut. A single layer of midgut columnar epithelial cells abuts on the basal lamina at one end with microvilli extending into the gut lumen at the other. Nidi of regenerative cells were observed between columnar epithelial cells at the basal lamina. Malpighian tubules are attached to the posterior end of the midgut. The hindgut consists of the pylorus, a muscular ileum connecting to a bulbous rectum, which then opens to the anus

Application of balsam fir sawfly nucleopolyhedrovirus against its natural host Neodiprion abietis (Hymenoptera Diprionidae

Abstract -Fifty-hectare blocks of balsam fir forest, in western Newfoundland Canada, were treated with 1 -3 x 10 9 occlusion bodies/hectare of Neodiprion abietis nucleopolyhedrovirus (NeabNPV) in 2.5 L 20% aqueous molasses using Cessna 188 'Ag Truck' airplanes equipped with Micronaire AU 4000 rotary atomizers. In the weeks following application, there was higher balsam fir sawfly larval mortality in the spray blocks than in the control. In the subsequent year, there was lower percentage egg hatch and higher larval mortality in samples collected from the spray blocks compared to those from the control block. Balsam fir sawfly pupae with white as opposed to brown pupal cases were significantly more like to harbour NeabNPV infection. These results suggest that aerial applications of NeabNPV can suppress outbreaking balsam fir sawfly populations

Comparative genome sequence analysis of Choristoneura occidentalis Freeman and C. rosaceana Harris (Lepidoptera: Tortricidae) alphabaculoviruses.

The complete genome sequences of Choristoneura occidentalis and C. rosaceana nucleopolyhedroviruses (ChocNPV and ChroNPV, respectively) (Baculoviridae: Alphabaculovirus) were determined and compared with each other and with those of other baculoviruses, including the genome of the closely related C. fumiferana NPV (CfMNPV). The ChocNPV genome was 128,446 bp in length (1147 bp smaller than that of CfMNPV), had a G+C content of 50.1%, and contained 148 open reading frames (ORFs). In comparison, the ChroNPV genome was 129,052 bp in length, had a G+C content of 48.6% and contained 149 ORFs. ChocNPV and ChroNPV shared 144 ORFs in common, and had a 77% sequence identity with each other and 96.5% and 77.8% sequence identity, respectively, with CfMNPV. Five homologous regions (hrs), with sequence similarities to those of CfMNPV, were identified in ChocNPV, whereas the ChroNPV genome contained three hrs featuring up to 14 repeats. Both genomes encoded three inhibitors of apoptosis (IAP-1, IAP-2, and IAP-3), as reported for CfMNPV, and the ChocNPV IAP-3 gene represented the most divergent functional region of this genome relative to CfMNPV. Two ORFs were unique to ChocNPV, and four were unique to ChroNPV. ChroNPV ORF chronpv38 is a eukaryotic initiation factor 5 (eIF-5) homolog that has also been identified in the C. occidentalis granulovirus (ChocGV) and is believed to be the product of horizontal gene transfer from the host. Based on levels of sequence identity and phylogenetic analysis, both ChocNPV and ChroNPV fall within group I alphabaculoviruses, where ChocNPV appears to be more closely related to CfMNPV than does ChroNPV. Our analyses suggest that it may be appropriate to consider ChocNPV and CfMNPV as variants of the same virus species