Long-term laboratory evaluation ofRogas lymantriae: A braconid endoparasite of the gypsy moth,Lymantria dispar

Rogas lymantriae Watanabe was reared onLymantria dispar (L.) for 125 generations in the laboratory following importation from Japan. No deleterious effects of colonization were observed and one measured parameter, successful parasite emergence, significantly improved over time. Decreased density of the host or conspecific female parasites resulted in higher levels of parasitism and numbers of females in the next generation. During 3 different years, colony production was significantly increased with little difficulty to provide parasites for inoculative releases in the field.      

Specificity of Developmental Resistance in Gypsy Moth （Lymantria dispar） to two DNA-Insect Viruses

Gypsy moth (Lymantria dispar) larvae displayed marked developmental resistance within an instar to L. dispar M nucleopolyhedrovirus (LdMNPV) regardless of the route of infection (oral or intrahemocoelic) in a previous study, indicating that in gypsy moth, this resistance has a systemic component. In this study, gypsy moth larvae challenged with the Amsacta moorei entomopoxvirus (AMEV) showed developmental resistance within the fourth instar to oral, but not intrahemocoelic, inoculation. In general, gypsy moth is considered refractory to oral challenge with AMEV, but in this study, 43% mortality occurred in newly molted fourth instars fed a dose of 5×10⁶ large spheroids of AMEV; large spheroids were found to be more infectious than small spheroids when separated by a sucrose gradient. Developmental resistance within the fourth instar was reflected by a 2-fold reduction in mortality (18%–21%) with 5×10⁶ large spheroids in larvae orally challenged at 24, 48 or 72 h post-molt. Fourth instars were highly sensitive to intrahemocoelic challenge with AMEV; 1PFU produced approximately 80% mortality regardless of age within the instar. These results indicate that in gypsy moth, systemic developmental resistance may be specific to LdMNPV, reflecting a co-evolutionary relationship between the baculovirus and its host.     

N-glycan structures of human transferrin produced by Lymantria dispar (gypsy moth) cells using the LdMNPV expression system

N-glycan structures of recombinant human serum transferrin (hTf) expressed by Lymantria dispar (gypsy moth) 652Y cells were determined. The gene encoding hTf was incorporated into a Lymantria dispar nucleopolyhedrovirus (LdMNPV) under the control of the polyhedrin promoter. This virus was then used to infect Ld652Y cells, and the recombinant protein was harvested at 120 h postinfection. N-glycans were released from the purified recombinant human serum transferrin and derivatized with 2-aminopyridine; the glycan structures were analyzed by a two-dimensional HPLC and MALDI-TOF MS. Structures of 11 glycans (88.8% of total N-glycans) were elucidated. The glycan analysis revealed that the most abundant glycans were Man1-3(+/-Fucalpha6)GlcNAc2 (75.5%) and GlcNAcMan3(+/-Fucalpha6)GlcNAc2 (7.4%). There was only approximately 6% of high-mannose type glycans identified. Nearly half (49.8%) of the total N-glycans contained alpha(1,6)-fucosylation on the Asn-linked GlcNAc residue. However alpha(1,3)-fucosylation on the same GlcNAc, often found in N-glycans produced by other insects and insect cells, was not detected. Inclusion of fetal bovine serum in culture media had little effect on the N-glycan structures of the recombinant human serum transferrin obtained.     

Phenology of Lymantria dispar (Lepidoptera: Lymantriidae), male flight and the effect of moth dispersal in heterogeneous landscapes

 A model of Lymantria dispar development was assembled from the published literature and used to predict the period of male moth flight in the United States. Model predictions were compared with observations made with pheromone traps in several locations throughout the United States but especially in Virginia, West Virginia and North Carolina between 1995 and 1996. The model was found to provide accurate and unbiased forecasts of the dates of 5%, 50% and 95% cumulative trap catch, particularly at lower elevations. In areas of high topographic diversity (such as West Virginia), deviations between model output and observations were minimized by basing predictions of 5% and 50% cumulative catch on minimum elevation within neighborhoods of 25–81 km². This model of L. dispar male flight phenology can be used to time the deployment and retrieval of pheromone traps in intensive or extensive monitoring programs. However, a better understanding of moth movement is needed to fully explain the patterns of local trap catch. Received: 9 October 1997/Accepted: 8 December 1997     

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.     

Mating-induced loss of sex pheromone and sexual receptivity in insects with emphasis on Helicoverpa zea and Lymantria dispar

Mating in most species of insects leads to a transient or permanent loss in sexual receptivity of the females. Among moths, this loss of receptivity is often accompanied with a loss of the sex pheromone in the absence of calling, which also could be temporary or permanent. Most of the earlier work on changes in reproductive behavior after mating was done with Diptera in which sperm and/or male accessory gland secretions were shown to be responsible for termination of receptivity. In the corn earworm moth, Helicoverpa zea, mated females become depleted of pheromone and become nonreceptive to further mating attempts, but only for the remainder of the night of mating. A pheromonostatic peptide isolated from the accessory glands of males may be responsible for the depletion of pheromone, while the termination of receptivity is independently controlled. In the gypsy moth, Lymantria dispar, the changes in behavior following mating are permanent. In this species, the switch from virgin to mated behavior involves three steps: a physical stimulation associated with mating, transfer of viable sperm to the spermatheca, and commencement of oviposition. Signals generated by these factors operate through neural pathways and, unlike in H. zea, accessory gland factors seem not to be involved. © 1994 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

Pheromone-mediated upwind flight of male gypsy moths, Lymantria dispar, in a forest

Abstract Lymantria dispar L. males flying upwind in a pheromone plume in a forest were video-recorded at 2.5, 10 and 20 m from the source of pheromone. Males flew slower and steered more across the wind as they approached the source. In concert, their ground speed decreased and track angles increased. In contrast to these changes, their drift angles were fairly constant and the transverse component of image flow, above and/or below the moths eyes, showed almost no change. The inter-turn duration (time between sequential turns), a temporal aspect of the male flight manoeuvres, showed a consistent but relatively small increase as the distance from the source increased. The flight tracks narrowed as the males approached close (2.5 m) to the source. Because of unpredicted correlations between physical variables (i.e. temperature, wind velocity) and the distance from the source, we used principal components analysis to generate a set of completely independent variables. Greater than 90% of the variability in the data could be explained by four principal factors which corresponded well with known relationships in the flight manoeuvres. All four of these factors showed a significant regression against distance to the source. Although uncontrolled factors such as temperature and wind velocity may have contributed to changes in flight behaviour, recent data indicate that, in addition to concentration, certain temporal and spatial characteristics (i.e. burst period, burst return period) of plumes in wind vary systematically with distance from the source. We propose that L.dispar males might adjust their flight manoeuvres in response to these changes.     

The effect of varying alkaloid concentrations on the feeding behavior of gypsy moth larvae, Lymantria dispar (L.) (Lepidoptera: Lymantriidae)

Nine alkaloids (acridine, aristolochic acid, atropine, berberine, caffeine, nicotine, scopolamine, sparteine, and strychnine) were evaluated as feeding deterrents for gypsy moth larvae (Lymantria dispar (L.); Lepidoptera: Lymantriidae). Our aim was to determine and compare the taste threshold concentrations, as well as the ED₅₀ values, of the nine alkaloids to determine their potency as feeding deterrents. The alkaloids were applied to disks cut from red oak leaves (Quercus rubra) (L.), a plant species highly favored by larvae of this polyphagous insect species. We used two-choice feeding bioassays to test a broad range of biologically relevant alkaloid concentrations spanning five logarithmic steps. We observed increasing feeding deterrent responses for all the alkaloids tested and found that the alkaloids tested exhibited different deterrency threshold concentrations ranging from 0.1 to 10 mM. In conclusion, it appears that this generalist insect species bears a relatively high sensitivity to these alkaloids, which confirms behavioral observations that it avoids foliage containing alkaloids. Berberine and aristolochic acid were found to have the lowest ED₅₀ values and were the most potent antifeedants. Handling Editor: Joseph Dickens.     

Influence of ambient and thoracic temperatures upon sexual behaviour of the gypsy moth, Lymantria dispar

ABSTRACT. In an ambient temperature ( T _a) range of 18–28°C, thoracic temperatures ( T _th) of individual male Lymantria dispar (L.), caught at flight in the field, ranged from 21 to 36.5°C, with a correlation coefficient of 0.63 between T _th and ambient temperature ( T _a). Ambient temperature (and insolation) altered the insect's body temperature and the probabilities, latencies, and durations of preflight responses to pheromone. In a wind tunnel at 16 and 20°C, quiescent males exposed to pheromone raised their T _th by sustained wing fanning from 17 and 21°C, respectively, to c. 24°C before takeoff. At 24 and 28°C ambient, T _th rose by takeoff to 28 and 31°C, respectively. The latencies of male wing fanning in response to pheromone decreased from 1.44 min at 16°C ambient, to 0.58 min at 20°C, to 0.26 min at 24°C, and to 0.16min at 28°C. The components of behaviour (antennal twitch, body jerk, step and wing tremor) that occurred between quiescence and wing fanning were more frequent at ambients of 16 and 20°C than at 24 and 28°C.     

Response of Lymantria dispar L. (Lepidoptera: Lymantriidae) to Bacillus thuringiensis subsp. kurstaki at different ingested doses and temperatures

We examined mortality and feeding inhibition response of Lymantria dispar L. (Lepidoptera: Lymantriidae) larvae to ingested doses of Bacillus thuringiensis subsp. kurstaki as a function of dose, instar and temperature. We developed generalized (logistic) linear mixed models and a mixture survival model, commonly used in medical statistics, to analyze the complex data set. We conducted bioassays of Foray 48B with larvae from the NJSS laboratory stock, using droplet imbibing or force-feeding to ensure dose ingestion. The dose causing mortality in 50% of the test population (LD₅₀) under standard test conditions (22 °C) ranged from 0.019 International Units (IU)/larva for first instar larvae (L₁) to 1.6 IU/larva for L₄. Temperature affected larval mortality in two ways. Mortality occurred sooner and progressed more rapidly with increasing temperature (13-25 °C) at each dose level and instar, while the maximum level of mortality attained by each instar decreased with increasing rearing temperature. The mechanisms underlying this effect are being investigated. Larvae that survived exposure to B. thuringiensis resumed feeding after a period that was dependent on instar, dose, and temperature. The equations describing observed mortality and feeding recovery responses were used to construct a simulation model, which was able to predict both processes, and which forms the basis for a process-oriented model that can be used as a decision support tool in aerial sprays.     

The effects of thinning and gypsy moth defoliation on wood volume growth in oaks

Stem dissection and dendroecological methods were used to examine the effects of thinning and defoliation by gypsy moth (Lymantria dispar L.) on wood volume increment in oaks (Quercus rubra L., Q. alba L., Q. prinus L.). A model was developed to evaluate radial volume increment growth at three time periods: before defoliation, during defoliation and after defoliation, as a function of species, defoliation intensity and crown position. Volume increment during these same time periods was also compared at different stem locations. Trees were defoliated for two consecutive years and results indicated that volume loss was greater during the second year of defoliation with complete recovery taking 2–3 years after defoliation. Oaks in thinned stands had similar reductions in annual volume increment during defoliation as those in the unthinned stand. Annual volume increment demonstrated a decreasing trend from stump to base of the live crown and volume increment of the lowest log (from stump height to 1.37 m), was always higher than upper log sections, even during defoliation. Both earlywood and latewood increments were reduced during defoliation; however, latewood reductions were distributed along entire stems while earlywood reductions were greater on upper stem sections within the crown.     

Development, fecundity and survival of the herbivore Lymantria dispar and the number of plant species in its diet

ABSTRACT.

• 1 Larval gypsy moth Lymantria dispar (L.) mortality depends upon the number and type of plant species consumed as well as the sequence in which they are consumed.
• 2 When a two-species diet is composed of a favourable and an unfavourable species, larval mortality is higher on the two-species diet than on the diet of the favourable species alone.
• 3 When a two species diet is composed of a favourable and unfavourable coniferous species, larval mortality is higher on the single conifer species diet than on the two-species diet.
• 4 Two-species diets, in which one species is a conifer, produce individuals with a greater fecundity than diets of either species alone.
• 5 The influence of diets composed of two favourable species, on development, size and fecundity, depends both on the relative quality of each of the two plant species and which of the two is consumed by older instars.

     

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.     

Identification and characterization of an ecdysiotropic peptide from brain extracts of the gypsy moth,Lymantria dispar

A peptide (Lymantria TE) was isolated from brains of the gypsy moth, Lymantria dispar, which stimulates synthesis of ecdysteroid in the testes of larval and pupal insects. This ecdysiotropic peptide was purified and its structure determined to be NH₂-IIe-Ser-Asp-Phe-Asp-Glu-Tyr-Glu-Pro-Leu-Asn-Asp-Ala-Asp-Asn-Asn-Glu-Val-Leu-Asp-Phe-OH using protein sequence analysis and electrospray mass spectrometry. The peptide was biphasic in activity, with maximal activity in the pupal testes at 10⁻¹³ M and 10⁻⁹ M, with a minimum at 10⁻¹⁰ M, and with maxima at 10⁻¹⁵ M and 10⁻¹⁰ M and minimum at 10⁻¹³ M for larval testes. Arch. Insect Biochem. Physiol. 34:175–189, 1997. © 1997 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

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.

     

Exploration into a mechanism of transgenerational transmission of nucleopolyhedrovirus in Lymantria dispar L. in Western Siberia

The mechanisms of nucleopolyhedrovirus (NPV)transmission through gypsy moth (Lymantriadispar L., Lepidoptera: Lymantriidae)populations in Western Siberia wereinvestigated. Field and laboratory studies werecarried out during 1992–1998 in a gypsy mothoutbreak in the Novosibirsk Area (WesternSiberia), Russia. The damaged trees were primarilytwo species of birch, Betula pendula Roth. and B. pubescensEhrh. Investigations were performed at threestages of the outbreak: population increase(1992–1993), population peak (1994–1995) andpopulation decline (1996–1998). Experiments onactivation of occult virus by different stressfactors were carried out in order to clarifythe mechanism of transgenerational transmissionof NPV. In both laboratory and fieldexperiments NPV-caused mortality levels werelow; most insects did not acquire a lethal doseof virus. Occult virus can provide an importantroute of transgenerational NPV transmission,particularly in Western Siberia where gypsymoths migrate by female flight and can moveaway from trees contaminated by virus fromprevious larval infections.     

Variation in temperature and dietary nitrogen affect performance of the gypsy moth (Lymantria dispar L.)

Abstract. . The independent and interactive effects of temperature and dietary nitrogen content on performance of the gypsy moth (Lymantria dispar L.) were examined. In long-term feeding trials, larvae were reared from egg hatch to pupation on low (1.5%) and high (3.7% dry weight) nitrogen diets, under three temperature regimes. Short-term feeding trials with fourth instars and the same treatments were conducted in order to calculate nutritional indices.
Higher temperatures did not influence larval survival and marginally increased final pupal weights, but strongly decreased long-term development rates. They also accelerated short-term growth and consumption rates, and tended to improve food processing efficiencies. High concentrations of dietary nitrogen increased survival rates and final pupal weights markedly, but decreased long-term development rates only marginally. A high content of dietary nitrogen also accelerated short-term development and growth rates, reduced consumption rates, and improved food digestibility. Insects responded to low nitrogen-content diets primarily by eating faster, rather than by altering efficiency of nitrogen use. In the short-term feeding trials, thermal regime and dietary nitrogen interacted to influence growth rates, overall food processing efficiencies and nitrogen consumption rates. No interactive effects were observed in long-term studies.
This research demonstrates that small changes in thermal regime and ecologically relevant variation in dietary nitrogen content can strongly affect gypsy moth performance. Moreover, various performance parameters are differentially sensitive to the direct and interactive effects of temperature and diet.     

Ion Channels Induced in Planar Lipid Bilayers by the Bacillus thuringiensis Toxin Cry1Aa in the Presence of Gypsy Moth (Lymantria dispar) Brush Border Membrane

The apical brush border membrane, the main target site of Bacillus thuringiensis toxins, was isolated from gypsy moth (Lymantria dispar) larval midguts and fused to artificial planar lipid bilayer membranes. Under asymmetrical N-methyl-d-glucamine-HCl conditions (450 mm cis/150 mm trans, pH 9.0), which significantly reduce endogenous channel activity, trypsin-activated Cry1Aa, a B. thuringiensis insecticidal protein active against the gypsy moth in vivo, induced a large increase in bilayer membrane conductance at much lower concentrations (1.1–2.15 nm) than in receptor-free bilayer membranes. At least 5 main single-channel transitions with conductances ranging from 85 to 420 pS were resolved. These Cry1Aa channels share similar ionic selectivity with P _Cl/P _NMDG permeability ratios ranging from 4 to 8. They show no evidence of current rectification. Analysis of the macroscopic current flowing through the composite bilayer suggested voltage-dependence of several channels. In comparison, the conductance of the pores formed by 100–500 nm Cry1Aa in receptor-free bilayer membranes was significantly smaller (about 8-fold) and their P _Cl/P _NMDG permeability ratios were also reduced (2- to 4-fold). This study provides a detailed demonstration that the target insect midgut brush border membrane material promotes considerably pore formation by a B. thuringiensis Cry toxin and that this interaction results in altered channel properties. Received: 23 February 2001/Revised: 15 June 2001     

Effects of transferring gypsy moth, Lymantria dispar, larvae between artificial diet and Quercus rubra foliage

Fourth instar gypsy moth larvae, Lymantria dispar, from a laboratory colony were transferred between artificial diet and three ages of leaves from a preferred tree, Quercus rubra. Mortality after transfers was generally minimal (<13.3%) and did not differ from mortality of larvae remaining on the same food. Larvae that were transferred to a new food accepted artificial diet and the early season foliage that is normally eaten by early instars as readily as larvae that were not transferred. Pupal weights from larvae continually fed artificial diet or transferred from artificial diet to early season foliage were not significantly different. For trials beginning during June and later, 12-day weight gain and pupal weight were greater for individuals transferred to or reared continually on artificial diet than for individuals transferred to or reared continually on mature foliage.First instars of wild larvae fed artificial diet gained less weight after 3 weeks than foliage-fed first instars. Pupal weights of wild larvae transferred from June foliage to artificial diet as fourth instars were not significantly different from pupal weights of wild larvae remaining on foliage. Thus, the response of laboratory colony larvae to artificial diet was superior to that of wild larvae.

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Résumé Des chenilles du quatrième stade de L. dispar, élevées au laboratoire sur régime artificiel et sur feuilles de différents stades de leur chêne préféré, Q. rubra, ont été transférées de l'un à l'autre de ces régimes alimentaires. La mortalité après transfert est généralement faible (<13,3%) et de même importance que pour les chenilles maintenues sur le régime artificiel. Les chenilles transférées sur un nouvel aliment ont accepté le régime artificiel ou le feuillage jeune,- normalement consommé par les premiers stades larvaires-, aussi facilement que les chenilles qui n'avaient pas été transférées. Les poids des chrysalides des chenilles élevées continuellement sur régime artificiel ou transférées sur feuillage jeune n'ont pas présenté de différences significatives. Pour des expériences commencées en juin ou ultérieurement, les gains de poids à 12 jours et pour les chrysalides sont plus élevés pour les insectes maintenus ou transférés sur régime artificiel que pour ceux transférés ou maintenus sur feuillage âgé.Le poids des chrysalides, provenant de chenilles de la nature tranférées au quatrième stade, d'un feuillage de juin à un régime artificiel, n'a pas différé significativement du poids des chrysalides provenant de chenilles maintenues sur feuillage. Les premiers stades de chenilles de la nature élevées sur régime artificiel ont pris moins de poids en trois semaines que les premiers stades ayant consommé du feuillage.

     

Release mechanism of sex pheromone in the female gypsy moth <Emphasis Type="Italic">Lymantria dispar</Emphasis>: a morpho-functional approach

A morpho-functional investigation of the sex pheromone-producing area was correlated with the pheromone release mechanism in the female gypsy moth Lymantria dispar. As assessed by male electroantennograms (EAG) and morphological observations, the pheromone gland consists of a single-layered epithelium both in the dorsal and ventral halves of the intersegmental membrane between the 8th and 9th abdominal segments. By using the male EAG as a biosensor of real-time release of sex pheromone from whole calling females, we found this process time coupled with extension movements of the ovipositor. Nevertheless, in females in which normal calling behavior was prevented, pheromone release was detected neither in absence nor in presence of electrical stimulation of the ventral nerve cord/terminal abdominal ganglion (TAG) complex. Tetramethylrhodamine-conjugated dextran amine stainings also confirm the lack of any innervation of the gland from nerves IV to VI emerging from the TAG. These findings indicate that the release of sex pheromone from the glands in female gypsy moths is independent of any neural control exerted by the TAG on the glands, at least by way of its three most caudally located pairs of nerves, and appears as a consequence of a squeezing mechanism in the pheromone-producing area.