Bioassay of a nucleopolyhedrosis virus of the gypsy moth, Porthetria dispar

The pathogenicity of an American isolate of the nucleopolyhedrosis virus of Porthetria dispar was studied. Laboratory data on third-instar larvae showed that mortality was directly related to virus concentration. The computed LD₅₀ was 1,729 PIBs/larva or 72 PIBs/mg larval body weight. The LT₅₀'s for 2.5 × 10⁶, 2.5 × 10⁵, 2.5 × 10⁴, 5 × 10³, and 2.5 × 10³ PIBs/larva were 8.1, 9.9, 11.3, 12.2, and 13.1 days, respectively. Approximately 37 and 60% of the total larval mortality occurred during the third- and fourth-instar, respectively. The periods to pupation and the pupal weights of survivors apparently were not affected by virus concentration. Moth emergence from surviving pupae was not reduced.     

Experimental mycosis of the gypsy moth,Porthetria dispar [Lep.: Lymantriidae] byEntomophthora virulenta,with notes on its mammalian safety

Larvae of the gypsy mothPorthetria dispar (L.) were infected with sporulating cultures of the fungus,Entomophthora virulenta Hall & Dunn using the spore shower technique. The infectivity of the parasite in the host and its histopathology was observed. Penetration of the integument was recorded at 24 hours post-inoculation and conidiophore production occurred at 72 hours after infection. Experiments on mammalian toxicity against laboratory mice showed no significant differences between control and treated mice in body weight, blood cell counts, food and liquid consumption and in the post-mortem examination.

---

Résumé Des larves dePorthetria dispar (L.) ont été infectées par des cultures en sporulation du champignon,Entomophthora virulenta Hall & Dunn par projection des spores à partir de la culture. L'infection par le pathogène et les processus histopathologiques ont été observés. La pénétration du tégument a été notée 24 heures après la contamination et les conidiophores apparaissent 72 heures après l'infection. Des essais sur souris ont montré qu'il n'y a pas de différences entre les animaux traités et les témoins pour le poids, les formules sanguines, la consommation d'eau et d'aliment et les examens post-mortem.

---

---

This study was supported by a grant from the Faculty Research Committee of Rhode Island College and was awarded jointly to the authors.     

Structure-activity studies with pheromone-binding proteins of the gypsy moth,Lymantria dispar

Pheromone olfaction in the gypsy moth, Lymantria dispar, involves accurate distinction of compounds with similar structure and polarity. The identified sex pheromone is (7R,8S)-2-methyl-7,8-epoxyoctadecane, 1a, and a known antagonist is (7Z)-2-methyloctadec-7-ene, 4a. The first step in pheromone olfaction is binding of odorants by small, soluble pheromone-binding proteins (PBPs), found in the pheromone-sensing hairs. We have studied the molecular determinants recognized by the two PBPs found in the gypsy moth, using three pheromone/PBP binding assays. Results indicate that (i) PBPs bind analogs of the pheromone with some discrimination; (ii) PBPs experience enhancement of binding when presented with 1a or its enantiomer and 4a simultaneously; and (iii) the binding enhancement is also seen at high ligand:PBP ratios. We found no evidence of allostery, so the synergistic binding effects and the concentration effect may only be explained by multimerization of PBPs with each other, which leads to more than one population of binding sites. We suggest that the enhanced ligand binding at high ligand:PBP ratios may serve to sequester excess ligand and thereby attenuate very strong signals.     

Development of male-incubated ovaries in the gypsy moth,Lymantria dispar

Ovaries from Lymantria dispar females were transplanted into an environment lacking vitellogenin, the male milieu, in order to determine how the presence of vitellogenin in the hemolymph affects the process of protein uptake by gypsy moth oocytes. When undeveloped ovaries from newly ecdysed last instar females were transplanted into males of the same stage, follicles detached from the germarium and increased in size, but the growth of oocytes proceeded more slowly than those from female controls. Although chorion fromation was delayed in male-grown ovaries, scanning electron microscopy of chorionated eggs recovered from adult males showed that a chorion with normal surface architecture was formed by the adult stage. SDS-PAGE analysis of the male-grown ovaries and hemolymph from males receiving ovaries showed that vitellogenin production was not stimulated by the organ transplant and only male hemolymph proteins were internalized by the male-incubated ovaries. Thus, in the absence of vitellogenin, endocytosis of male hemolymph proteins occurred, but the rate of oocyte growth was slowed.     

Soluble cuticular phenoloxidase in the gypsy moth Lymantria dispar

The soluble enzyme phenoloxidase (tyrosinase) from the larval cuticle of Lymantria dispar has been partially purified using Ultrogel ACA 34, and the activity has been determined using phenolic substrates. The enzyme exhibited more activity toward O-diphenolic substrates and monophenolic substrates. The enzyme is inhibited by diethyl dithiocarbamate, phenylthiourea, and thiourea. The enzyme has been localized in the 7% slab and disc PAGE as an intense band. The enzyme is suggested to be involved in wound healing. © 1992 Wiley-Liss, Inc.     

Ultrastructural studies of male-incubated ovaries of the gypsy moth,Lymantria dispar

Ovaries from Lymantria dispar females were transplanted into an environment lacking the vitellogenin ligand; i.e., the male milieu. Transmission electron micrographs comparing the terminal oocytes of male-grown ovaries and normal ovaries showed that yolk sphere diameters were reduced in the male-grown oocytes. However, there were larger numbers of these small yolk spheres per unit area of cytoplasm, indicating that the coalescence of endosomes into yolk spheres is reduced in the absence of vitellogenin. Although there are larger numbers of yolk spheres in male-grown oocytes, the smaller diameter of yolk spheres resulted in less area being taken up by yolk spheres per unit area of cytoplasm in male-grown oocytes, yielding lowered yolk production. This lowered yolk production is a result at least in part of the lowered number of coated vesicles per unit area of submembrane space and in part of the reduced interfollicular spaces seen in male-grown ovaries.     

Vertical transmission and overwintering of microsporidia in the gypsy moth, Lymantria dispar

Vertical transmission and the overwintering success of three different microsporidia infecting Lymantria dispar (Lepidoptera: Lymantriidae) larvae were investigated. Endoreticulatus schubergi, a midgut pathogen, was transmitted to offspring via female and male via the egg chorion (transovum transmission). Between 8% and 29% of the emerging larvae became infected. No spores of E. schubergi were found in surface-washed eggs. Nosema lymantriae, a microsporidium that causes systemic infections, was transovarially transmitted. Between 35% and 72% of the progeny were infected. Vairimorpha disparis, a fat body pathogen, was not vertically transmitted. The infectivity of spores that overwintered in cadavers of infected L. dispar varied by species, placement in the environment, and weather conditions. Spores of E. schubergi were still infective after an eight month exposure period of cadavers on the ground. Spores of N. lymantriae and V. disparis remained highly infective only when cadavers overwintered under a more or less continuous snow cover for four months.     

Physico-chemical characteristics of the DNA of the nuclear polyhedrosis virus of the moth Porthetria dispar L

DNA preparations were obtained after dissolving the inclusion bodies, polyhedra virus particles, from the purified bundle virus of Porthetria dispar L. nuclear polyhedrosis. The DNA molecules in the preparations obtained are of different conformation and separate within the CsCl density gradient in the presence of ethidium bromide into supercoiled catenated and relaxed circular molecules (with the admixture of linear molecules). The circular DNA was studied by electron microscopy. The size of virus genome according to the data of reassociation kinetics of DNA is about 100 MD. Estimated on the basis of the values of buoyant density (p) and the melting temperature (Tmelt.) the content of guanine-cytosine pairs (GC pairs) in the viral DNA varies from 61 up to 65 mol%, and in the insect cell DNA--from 38 up to 40 mol%. The viral and cellular DNA are distinctly separated by centrifugation within the CsCl density gradient.     

Underestimated mitochondrial diversity in gypsy moth Lymantria dispar from Asia

相似文献   

Male death resulting from hybridization between subspecies of the gypsy moth,Lymantria dispar

We explored the origin of all-female broods resulting from male death in a Hokkaido population of Lymantria dispar through genetic crosses based on the earlier experiments done by Goldschmidt and by testing for the presence of endosymbionts that are known to cause male killing in some insect species. The mitochondrial DNA haplotypes of the all-female broods in Hokkaido were different from those of normal Hokkaido females and were the same as those widely distributed in Asia, including Tokyo (TK). Goldschmidt obtained all-female broods through backcrossing, that is, F1 females obtained by a cross between TK females (L. dispar japonica) and Hokkaido males (L. dispar praeterea) mated with Hokkaido males. He also obtained all-male broods by mating Hokkaido females with TK males. Goldschmidt inferred that female- and male-determining factors were weakest in the Hokkaido subspecies and stronger in the Honshu (TK) subspecies. According to his theory, the females of all-female broods mated with Honshu males should produce normal sex-ratio broods, whereas weaker Hokkaido sexes would be expected to disappear in F1 or F2 generations after crossing with the Honshu subspecies. We confirmed both of Goldschmidt''s results: in the case of all-female broods mated with Honshu males, normal sex-ratio broods were produced, but we obtained only all-female broods in the Goldschmidt backcross and obtained an all-male brood in the F1 generation of a Hokkaido female crossed with a TK male. We found no endosymbionts in all-female broods by 4,′6-diamidino-2-phenylindole (DAPI) staining. Therefore, the all-female broods observed in L. dispar are caused by some incompatibilities between Honshu and Hokkaido subspecies.     

亚洲型舞毒蛾在北美的适生性

采用DYMEX V2.0软件和ArcGIS分析工具相结合的方法,提出亚洲型舞毒蛾Lymantria dispar(L.)的DYMEX参数指标体系和适生性评判标准,分析亚洲型舞毒蛾在北美的适生范围与适生程度。研究表明,加拿大的南部、美国的大部分地区以及墨西哥中南部极少部分区域为该虫的适生区。研究结果将为国家植物保护部门提供有关亚洲型舞毒蛾的植物检疫决策支持。     

Estimating the susceptibility of tree species to attack by the gypsy moth, Lymantria dispar

ABSTRACT.

• 1 Numbers of gypsy moth larvae feeding on each of 922 randomly sampled trees in a Quercus—Acer—Fraxinus forest in southwestern Quebec, Canada were counted in 1979 and in 1980 to quantify the larval feeding preferences as observed in the field for eighteen deciduous and one coniferous tree species at the northern range limit of the gypsy moth.
• 2 Both the diameter at breast height (dbh) and the estimated foliage biomass of the sampled trees were used to calculate the relative proportions of foliage represented by each of the nineteen tree species in the forest canopy. With these data on availability and utilization of the tree species by the gypsy moth larvae an Ivlev-type electivity index was used to quantify the larval feeding preferences. These preferences observed in the field define the susceptibility of a tree species to attack by the gypsy moth.
• 3 The feeding preferences calculated using estimated foliage biomass were comparable to the simpler calculation based on dbh (Spearman's rho = 0.79; P= 0.0001). The dbh-based feeding preferences remained almost unchanged in 1979 and 1980 (Spearman's rho = 0.83; P= 0.0001).
• 4 The composite 1979—80, dbh-based feeding preferences show Quercus rubra, Populus grandidentata, Ostrya virginiana, Amelanchier spp. and Acer saccharum were preferentially attacked by gypsy moth. Prunus serotina, Betula lutea, Acer rubrum, A. pensylvanicum, Fraxinus americana, Ulmus rubra, P. pensylvanicum and B. papyrifera were avoided. All nineteen tree species were, however, utilized to at least some degree by gypsy moth larvae.
• 5 These results quantitatively affirm and clarify earlier reports of gypsy moth feeding preferences in North America and Eurasia. The advantages and limitations of using an electivity index to estimate the susceptibility of different tree species to attack by folivores like the gypsy moth are discussed.

     

Characterisation of three Alphabaculovirus isolates from the gypsy moth,Lymantria dispar dispar (Lepidoptera: Erebidae), in Turkey

In this study, Lymantria dispar dispar larvae, collected from three different localities in Turkey, were examined for the presence of inclusion bodies under phase contrast and electron microscopes. Inclusion bodies from infected larvae were subjected to polymerase chain reaction using the conserved primers for polyhedrin (polh), late expression factor 8 (lef-8) and late expression factor 9 (lef-9) genes. Sequence analysis confirmed that larvae collected from the three different localities contained multiple nucleopolyhedrosis viruses (MNPVs). These isolates were designated LdMNPV-T1, LdMNPV-T2 and LdMNPV-T3. Phylogenetic analyses of these isolates were performed using target genes polh, lef-8 and lef-9. Restriction endonuclease analysis of the three geographic isolates with EcoRI and PstI enzymes demonstrated some differences existed among the isolates. According to the EcoRI profile, the mean estimated size for the complete genome of each isolate (LdMNPV-T1, LdMNPV-T2 and LdMNPV-T3) was calculated to be approximately 170, 153 and 170?kb, respectively. Insecticidal activities of each isolate were tested on L. d. dispar larvae using four different viral concentrations between 10³ and 10⁶?OBs/ml. Results showed that the mortalities for LdMNPV-T1, -T2 and -T3 ranged between 13–53%, 47–100% and 46–93%, respectively. The LC₅₀ and LC₉₅ values of LdMNPV-T2 were not significantly different from the respective corresponding values of the other two isolates. However, isolate LdMNPV-T2 killed larvae with a LC₅₀ value that was lower than the other two isolates. Our results suggested there are promising LdMNPV isolates in Turkey that can be used for microbial control of L. d. dispar larvae.     

Identification of 20-hydroxyecdysone and ecdysone from the pupa of the gypsy moth,Lymantria dispar

Normal and reverse-phase high-performance liquid chromatography in conjunction with radioimmunoassay and mass spectrometry were used to identify the free and conjugated ecdysteroids (after enzymatic hydrolysis) from day-4 pupae of the gypsy moth, Lymantria dispar L. Seven ecdysteroids were searched for, but only 20-hydroxyecdysone (964 ng/g fresh weight) and ecdysone (367 ng/g fresh weight) were detected. Analysis of conjugated ecdysteriods after liberation by hydrolysis also indicated the presence of 20-hydroxyecdysone (21.6 ng/g fresh weight) and ecdysone (2.4 ng/g fresh weight). Neither 26-hydroxyecdysone nor the 3α-epimers of 20-hydroxyecdysone or ecdysone were detected.     

Cuticle hydrolysis in four medically important fly species by enzymes of the entomopathogenic fungus Conidiobolus coronatus

Entomopathogenic fungi infect insects via penetration through the cuticle, which varies remarkably in chemical composition across species and life stages. Fungal infection involves the production of enzymes that hydrolyse cuticular proteins, chitin and lipids. Host specificity is associated with fungus–cuticle interactions related to substrate utilization and resistance to host‐specific inhibitors. The soil fungus Conidiobolus coronatus (Constantin) (Entomophthorales: Ancylistaceae) shows virulence against susceptible species. The larvae and pupae of Calliphora vicina (Robineau‐Desvoidy) (Diptera: Calliphoridae), Calliphora vomitoria (Linnaeus), Lucilia sericata (Meigen) (Diptera: Calliphoridae) and Musca domestica (Linnaeus) (Diptera: Muscidae) are resistant, but adults exposed to C. coronatus quickly perish. Fungus was cultivated for 3 weeks in a minimal medium. Cell‐free filtrate, for which activity of elastase, N‐acetylglucosaminidase, chitobiosidase and lipase was determined, was used for in vitro hydrolysis of the cuticle from larvae, puparia and adults. Amounts of amino acids, N‐glucosamine and fatty acids released were measured after 8 h of incubation. The effectiveness of fungal enzymes was correlated with concentrations of compounds detected in the cuticles of tested insects. Positive correlations suggest compounds used by the fungus as nutrients, whereas negative correlations may indicate compounds responsible for insect resistance. Adult deaths result from the ingestion of conidia or fungal excretions.     

Neurological influences on pheromone release and calling behaviour in the gypsy moth, Lymantria dispar

ABSTRACT. Surgical removal of the brain or disconnection of the last abdominal ganglion from the ventral nerve cord prevented sex pheromone release in female Lymantria dispar (L.) (Lymantriidae), as assayed by the male wing-fanning response. The calling behaviour continued to occur in individuals whose terminal abdominal ganglion had been thus isolated, however, indicating that the neural mechanisms controlling calling function independently in the last abdominal ganglion.