The alimentary canal ofNeodiprion abietislarvae 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.
The Neodiprion abietis Gammabaculovirus (Baculoviridae: NeabNPV) is virulent, highly contagious, and infects only midgut epithelial cells of balsam fir sawfly larvae, but infections can carry through to adult sawflies in the midgut. Larval infections are characterized by hypertrophy of midgut epithelial cell nuclei, where virogenic stromata develop to produce nucleocapsids that are singly enveloped before occlusion into occlusion bodies. Infected, occlusion body-laden cells slough from the midgut epithelium as a result of a dissolution of the basal lamina. Infected cells undergo lysis, and viral occlusion bodies exit affected larvae in a watery diarrhea to infect other balsam fir sawfly larvae. A budded virus stage was not observed, but nucleocapsid and occlusion body formation resembled the development of occlusion-derived virions and occlusion bodies in lepidopteran alphabaculoviruses.
Infections of two heterologous insect cell lines derived from Malacosoma disstria (Md108) and Choristoneura fumiferana (Cf70) by the Lambdina fiscellaria lugubrosa nucleopolyhedrovirus (LafiNPV-W) were characterized. Cytopathic effects characteristic of LafiNPV-W infection, including rounding of cells, nuclear hypertrophy, and occlusion body (OB) production, were observed in both cell lines. Budded virus titers were slightly higher in Md108 cells than Cf70 cells (5.8 x 10(7) versus 3.1 x 10(7) TCID(50) units mL(-1)). Viral replication kinetics and cytopathic effects induced by LafiNPV-W infection were very similar in both cell lines. Actin rearrangements and redistribution of heterochromatin and euchromatin were observed within 24 h post-inoculation (hpi), and large quantities of nucleocapsids and virions were observed by electron microscopy at 48 hpi in both cell lines. Cf70 cultures produced OBs with numerous embedded virions, while OBs in Md108 cultures contained few virions or were empty with nucleocapsids packed in the nucleoplasm between OBs. In bioassays against second instar L. fiscellaria lugubrosa, OBs derived from LafiNPV-W-infected Md108 cells induced significantly lower levels of mortality than OBs derived from LafiNPV-W-infected Cf70 cells or from infected L. fiscellaria fiscellaria larvae.
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