Bioassay of a purified nuclear polyhedrosis virus against Heliothis armigera

A precise bioassay method, which is not limited by lack of field applicability, as are peroral administration techniques, is described. Purified nuclear polyhedrosis virus (NPV) suspensions were assayed against third and fourth instar Heliothis armigera larvae to provide standards for additive and field testing. Third instar larvae proved to be approximately one hundred times more susceptible to the NPV disease than fourth instar larvae. The minimum time to mortality was 4 days.     

Interaction of Heliothis armigera nuclear polyhedrosis viral capsid protein with its host actin

In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement.     

Effects of nuclear polyhedrosis virus on cannibalism in Helicoverpa armigera

在室内研究核型多角体病毒(HaSNPV)感染对棉铃虫Helicoverpa armigera(Hübner)幼虫同类相残行为的影响.结果显示:感病棉铃虫随感病程度的加重,越容易被健康棉铃虫残食,而自然死亡的感病棉铃虫、冻死的感病棉铃虫和冻死的健康棉铃虫三者被健康棉铃虫残食的百分率无显著差异.表明感病棉铃虫和病虫尸体更易于被健康棉铃虫残食,是由于棉铃虫体力减弱而失去反击能力,不是由于病毒本身的影响.以健康棉铃虫、感病棉铃虫为残食者,冻死的病虫为被残食者,相残率无显著差异,表明病毒并未改变棉铃虫残食同类的天性.残食病虫的健康棉铃虫的化蛹率和羽化率均低于正常的健康棉铃虫,残食者为相残行为付出了很高的代价.     

Effect of dew from cotton and soybean foliage on activity of Heliothis nuclear polyhedrosis virus

The ionic composition of dew collected from foliage of cotton and soybean plants as well as its potential for inactivation of Heliothis NPV were compared. Cotton dew had a mean pH of 8.8 and was more alkaline than soybean dew (pH 7.8). Cotton dew had a higher concentration of ions that did soybean dew but there was no qualitative difference in the cation content of dew from the two hosts. In bioassay tests, no loss of activity occurred when polyhedra were held in dew of either plant species. If the dew in which polyhedra were suspended was air-dried and resuspended daily in deionized water, polyhedra in soybean dew remained active but in cotton dew retained little activity after 7 days. Also, electron microscopical examination of polyhedra pelleted from these cotton dew preparations showed much dissolution after 7 days. Although there was dissolution of polyhedra in cotton dew when dried, an examination of polyhedra on the upper surface of either plant species in the field showed little degradation after 7 days.     

核型多角体病毒感染对棉铃虫幼虫同类相残行为的影响

在室内研究核型多角体病毒（HaSNPV）感染对棉铃虫Helicoverpa armigera（Htibner）幼虫同类相残行为的影响。结果显示：感病棉铃虫随感病程度的加重,越容易被健康棉铃虫残食,而自然死亡的感病棉铃虫、冻死的感病棉铃虫和冻死的健康棉铃虫三者被健康棉铃虫残食的百分率无显著差异。表明感病棉铃虫和病虫尸体更易于被健康棉铃虫残食,是由于棉铃虫体力减弱而失去反击能力,不是由于病毒本身的影响。以健康棉铃虫、感病棉铃虫为残食者,冻死的病虫为被残食者,相残率无显著差异,表明病毒并未改变棉铃虫残食同类的天性。残食病虫的健康棉铃虫的化蛹率和羽化率均低于正常的健康棉铃虫,残食者为相残行为付出了很高的代价。     

Susceptibility of Heliothis armiger to a commercial nuclear polyhedrosis virus

The susceptibility of Heliothis armiger larvae of different ages to a commercial nuclear polyhedrosis virus (NPV), Elcar, was determined by bioassay. The median lethal dosage (LD₅₀) increased 150-fold during the first week of larval life at 25°C, i.e., during development to early fourth instar, but daily feeding rate and thus potential virus acquisition also increased. A linear relationship was determined between log LD₅₀ and larval length, indicating that larval length constitutes a useful index for estimating the susceptibility of larval populations. Median lethal times (LT₅₀s) were similar for larvae tested at ages of 0 to 7 days and ranged from 3.6 to 8.0 days at 30°C. The amount of virus produced in a single, infected neonate was equivalent to 1.4 × 10⁶ LD₅₀s for neonates, a 900,000-fold increase on the dose supplied. The data support the practice of directing the NPV against neonates, but, on the basis of larval susceptibility alone, the age of larvae at treatment may not always be critical.     

Replication of Heliothis zea nuclear polyhedrosis virus in cloned cell lines

Clones from two Heliothis zea (Lepidoptera:Noctuidae) ovarian cell lines, BCIRL-Hz-AM1 (AM1) and BCIRL-Hz-AM3 (AM3), were generated and their ability to produce H. zea nuclear polyhedrosis virus (HzSNPV) was compared. Titers of extracellular virus (ECV) ranged from 5.5 (AM3-F9) to 44.9 x 10(5) PFU/ml (AM1-A4), with the parental cell lines AM3 and AM1 producing 14.8 and 26.4 x 10(5) PFU/ml, respectively. Concentrations of polyhedral inclusion bodies (PIB) produced by the cloned lines ranged from 0.7 (AM3-F9) to 59.6 x 10(6) PIB/ml (AM1-B3), while the parental cell lines generated 6.5 (AM3) and 12.9 x 10(6) PIB/ml (AM1). The percentage of cells from the cloned lines that produced PIB ranged from 39 to 86.4% for AM3-F9 and AM1-A7, respectively, with the parental lines exhibiting 49.1% (AM3) and 75.3% (AM1) cells with PIB. The number of PIB per cell also differed markedly between cell lines, varying from 18.3 (AM3-F9) to 184.4 (AM1-B3) PIB/cell. The parental lines produced 57.3 (AM3) and 75.9 (AM1) PIB/cell. Thus, significant differences were seen in virus production (ECV, PIB) between parental cell lines, as well as between parental cell lines and their clones. In addition, cell lines were characterized with regard to their growth rates and isoenzyme patterns.     

Detection of nuclear polyhedrosis virus infection in Heliothis spp. by agar gel double diffusion

Nuclear Polyhedrosis virus infection was detected in single Heliothis zea larvae by the agar gel double diffusion technique using antiserum to alkali-solubilized polyhedra. Virus bands were observed in 2nd-5th stadium larvae following homogenization in 0.1 M Na₂CO₃-0.05 M NaCl, pH 11.0, in approximately one-half the time necessary for mortality to occur. In a field test, virus infection was detected by this method as early as 3 days after virus treatment.     

Effect of two granulosis viruses on the activity of the gypsy moth (Lepidoptera: Lymantriidae) nuclear polyhedrosis virus

Two granulosis viruses (GV) were tested as enhancers for the gypsy moth nuclear polyhedrosis virus (LdMNPV). Helicoverpa armigera (Hübner) CV (HaGV) had no detrimental effect upon larval growth and development, but in combination with LdMNPV it reduced both the LC50 and the LT50 for the NPV. In addition, the combination also adversely affected the growth and development of gypsy moth larvae. The LC50 of LdMNPV was reduced by as much as 300-fold (HaGV at 10(-2) dilution) and the LT50 was reduced by as much as 18% (HaGV at 10(-2) dilution). Spodoptera frugiperda (J. E. Smith) GV reduced the LC50 of LdMNPV by as much as 13-fold but had no effect upon the LT50.     

Relative susceptibility of several noctuid species to a nuclear polyhedrosis virus from Heliothis armiger

Several alternate hosts were tested for their relative susceptibility to an isolate of Galleria mellonella nuclear polyhedrosis virus. Neonate Trichoplusia ni, Heliothis zea, and Manduca sexta were all susceptible to per oral administration of purified polyhedra. Of the three alternate species tested, T. ni was the most susceptible, and exhibited the most variable mortality response over the dose range tested, while M. sexta was the least susceptible. We believe this represents the first report of a lethal virus infection in a sphingid species, and useful parameters for the successful inoculation of alternate hosts are discussed.     

Effect of larval age and dosage of nuclear polyhedrosis virus on the susceptibility of diamondback moth, Plutella xylostella

A laboratory experiment was conducted to study the efficacy of nuclear polyhedrosis virus (NPV) with nine concentrations against all stadia of Plutella xylostella (L). The susceptibility of the larvae was correlated negatively with the period of development of the larvae and positively with the virus concentrations. The highest mortality of 84% was recorded in first stadium larvae compared to lowest mortality of 38% in fourth stadium larvae. The LC₅₀ was 5.5×10¹ and 4.0×10⁴ polyhedral inclusion bodies (PIB)/ml for first and fourth stadium larvae, respectively.