In vitro and in vivo host range of Anticarsia Gemmatalis multiple nuclear polyhedrosis virus

Abstract

A clone of the wild type (wt) Anticarsia gemmatalis multiple nuclear polyhedrosis virus AgMNPV, derived from a geographical isolate (Hondrina, Brazil) and designated AgMNPV-CL4-3A1, was used to determine the host range of this virus in six established lepidopteran cell lines: Anticarsia gemmatalis (BCIRL-AG-AM1), Helicoverpa zea (BCIRL-HZ-AM1), Heliothis virescens (BCIRL-HV-AM1), Helicoverpa armigera (BCIRL-HA-AM1), Trichoplusia ni (TN-CL1), Bombyx mori (BMN), and a coleopteran cell line Anthonomus grandis (BRL-AG-1). In addition, the in vivo host range of this clone was also assayed in larvae of Helicoverpa zea, Heliothis virescens, Trichoplusia ni, and the homologous species Anticarsia gemmatalis by probit analysis. On the basis of temporal studies of TCID50 values, BCIRL-HV-AM1 cells gave the highest extracellular virus (ECV) titer (9.7 x 10(6) TCID50/ml) followed by BCIRL-HA-AM1 cells (8.3 x 10(5) TCID50/ml) and BCIRL-AG-AM1 cells (3.2 x 10(5) TCID50/ml). In addition, a low ECV titer of 1.37 x 10(3) TCID50/ml was detected from TN-CL1 cells 96 h postinoculation, while BRL-AG-1, BMN, and BCIRL-HZ-AM1 cells were nonpermissive to AgMNPV-CL4-3A1 on the basis of TCID50 results. AgMNPV-CL4-3A1 and the wild type AgMNPV had similar restriction profiles that were different from wild type AcMNPV. The LC50 values were 96.9, 564.6, 733.3, and 1.1 x 10(4) occlusion bodies/cm2 of diet for A. gemmatalis, Helicoverpa zea, Heliothis virescens, and T. ni, respectively.

Growth, metabolism and baculovirus production in suspension cultures of an Anticarsia gemmatalis cell line

“…Rodas et al (2005) reported to achieve a specific yield of 27.60 AgMNPV PIBs per cell in suspension cultures of the Sf9 cell line in Sf900 II serum-free medium, while Batista et al (2005) obtained a volumetric yield of 1.60 · 10 7 AgMNPV PIBs ml -1 in suspension cultures of the same cell line infected in a serum-supplemented medium at a cell density of 1 · 10 6 cells ml -1 (maximum specific yield of 16 AgMNPV PIBs per cell). Grasela and McIntosh (1998) assayed the production of AgMNPV PIBs in static cultures of the homologous cell line BCIRL-AG-AM1 in serum-supplemented TC-100 medium, obtaining a maximum specific yield of 14.20 AgMNPV PIBs per cell. Zhang et al (2005), infecting suspension cultures of the IPLBSf-21 cell line in TC-100 medium supplemented with 10% fetal calf serum, obtained a maximum volumetric yield of 1.10 · 10 6 PIBs ml -1 in cultures infected at an initial cell density of 4 · 10 5 cells ml -1 (maximum specific yield of 2.75 AgMNPV PIBs per cell).…”

“…Several cell lines have been evaluated for their ability to support AgMNPV replication (Castro et al 1997;Claus et al 1993;Grasela and McIntosh 1998;Lynn 2003;Rodas et al 2005;Sieburth and Maruniak 1988b). These reports show that the UFL-AG-286 homologous cell line (Sieburth and Maruniak 1988a) exhibits a higher susceptibility to AgMNPV infection, as well as a larger capability to replicate the virus, than other insect cell lines.…”

Morphological characterization of Anticarsia gemmatalis M nucleopolyhedrovirus infection in haemocytes from its natural larval host, the velvet bean caterpillar Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae)

“…AgMNPV is a rod-shaped, enveloped virus with circular, double-stranded, 133-kbp DNA genome, pathogenic to lepidopterans ( Johnson andMaruniak, 1989 andGrasela andMcIntosh, 1998). Like other baculoviruses from the Nucleopolyhedrovirus genus, it presents two phenotypes: budded viruses (BVs), which establish systemic infection; and occlusionderived viruses (ODVs), immersed in a proteinaceous matrix, constituting occlusion bodies (OBs) or polyhedra.…”

Establishment of a monarch butterfly (Danaus plexippus, Lepidoptera: Danaidae) cell line and its susceptibility to insect viruses

“…BCIRL-HsAM1 cell line was selected for the study of the replication of AcMNPV-HPP, PxMNPV and AfMNPV since these baculoviruses grow well in this cell line (McIntosh and Ignoffo, 1989;Kariuki et al, 2000). Similarly BCIRL-HvAM1 and BCIRL-HzAM1 were selected to study the replication of AgMNPV and HzSNPV respectively Grasela and McIntosh, 1998).…”