Auditory characteristics and sexual dimorphism in the gypsy moth

ABSTRACT. The auditory characteristics of two populations (laboratory reared and wild) of North American gypsy moths (Lymantriidae: Lymantria dispar L.) were sampled and the neurally derived thresholds of wild males and females to frequencies from 5 to 150 kHz compared. The noctuoid auditory receptors, Al and A2-cell, and putative proprioceptor, B-cell, were identified. Both sexes possess neurally responsive ears but females exhibit median best frequencies significantly lower than those of males. Audiogram comparisons reveal significantly different thresholds at 5–15 kHz, 30–120 kHz and 130–140 kHz, with females less sensitive to all but the lowest frequencies. Wild male populations reveal less audiogram variability than laboratory-reared individuals, while females' tuning curves appear more similar. The high variability present in colony moths warrants caution in the use of laboratory-reared insects for studies that assume natural levels of selection pressure. We suggest that male L. dispar possess adaptively functional ears tuned to the frequencies in the echo-location signals of bats but that the flightless females of this species are not exposed to bat predation and therefore possess ears in a state of evolutionary degeneration.     

Effects of microhabitat,time of day,and weather on predation of gypsy moth larvae

Summary Wire cages with different-sized meshes were placed on trunks and around leaves at different heights in oak trees and in forest litter. Gypsy moth, Lymantria dispar, instars II–V tethered with threads were placed in each cage (instars II–III only in leaf cages) as well as outside the cages. Predation of larvae decreased from near ground to mid-crown in trees and was highest in litter and very low on leaves. Predation in litter was not strongly related to cage type, suggesting that small, invertebrate predators were active there, but IV–V instars on trunks were mainly killed by relatively large predators, probably forest mice and shrews. Influences of the time of day and weather on predation were evaluated by observing tethered V–VI instars in litter and on tree trunks hourly. Ants, particularly carpenter ants (Camponotus ferrugineus) and Formica sp., and probably vertebrates, were conspicuous predators in the litter. Ants were most active at lower relative humidities, while other predators were apparently not influenced by humidity. No daily activity rhythms of predation were noted. Invertebrates appear to be important predators of larvae only in the litter whereas vertebrates, such as forest mice and shrews, also readily attack larvae on tree trunks.     

Carbon and nitrogen mineralization from decomposing gypsy moth frass

Defoliation of forests by insects is often assumed to produce a pulse of available nitrogen (N) from the decomposition of frass pellets. In this study we measured rates of carbon (C) and N mineralization from gypsy moth frass incubated with and without soil, and for soil alone. Incubations were at constant temperature and soil moisture conditions and lasted for 120 days. We found that gypsy moth frass contains much labile C as well as extractable N, and that the stimulation of microbial growth by the labile C results in immobilization of essentially all of the extractable N in the frass. The response of the microbes is fast, beginning within 1 day and lasting at least 90 days. This immobilization response represents an efficient mechanism for conserving N within a forest ecosystem after a defoliation event.     

Caterpillar guts and ammonia volatilization: retention of nitrogen by gypsy moth larvae consuming oak foliage

The gypsy moth (Lymantria dispar L.), a major defoliator of hardwood forests in the eastern U.S., has a highly alkaline midgut pH. We hypothesized that the high pH would cause high rates of ammonia (NH₃) volatilization as larvae consumed foliage, leading to potentially large losses of N from the ecosystem to the atmosphere during gypsy moth outbreaks. We measured NH₃ emission during the consumption of oak foliage by larvae in the laboratory. Surprisingly, we found very low amounts of NH₃ release of about 0.1% of the N consumed in foliage. We speculate that digestive mechanisms may limit NH₃ production in the midgut, and that the acidic environment of the hindgut traps most of the small amount of NH₃ that is produced, effectively preventing a potentially very large N loss from both larvae and ecosystem. The estimated rate of NH₃ emission from a defoliated forest is small compared to other inputs and outputs of N from the ecosystem, but could potentially enhance the neutralization of atmospheric acidity during the defoliation period. Received: 12 May 1998 / Accepted: 28 July 1998     

Stabilization of altitude and speed in tethered flying gypsy moth males: influence of (+) and (-)-disparlure

ABSTRACT. Changes in lift and thrust were elicited in tethered male gypsy moths, Lymantria dispar L. (Lepidoptera, Lymantriidae), by visual pattern elements moving radially either towards or from the point directly beneath their body, if the sex-pheromone, (+)-disparlure, was present. The sign of these changes was such as to counteract the pattern movements, which were generated by a rotating spiral beneath the moth. By restricting the area of spiral visible to the moth to either transverse or longitudinal sectors, flight altitude was affected by the centrifugal/centripetal movements in the lateral sectors, whereas flight speed was affected by those in the frontal sector. It is deduced that in free flight these compensatory reactions are responsible for the stabilization of flight altitude and speed, respectively. Surprisingly, without pheromone present these responses were usually not detectable: a wide range of flight altitude and speed was then observed. In the presence of (+)-disparlure, however, these responses were always strongly pronounced, the animal keeping within a narrow range of speed and altitude. These compensatory reactions were blocked by the attraction-inhibiting (-)-disparlure if presented in racemic mixture with the (+) form: the range of speed and altitude shown by the moth was then the same as without any pheromone. Under closed-loop conditions, the mean flight speed was reduced by the racemic mixture as well as by (+)-disparlure alone, however.     

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.     

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.      

Intraspecific variation in aspen phytochemistry: effects on performance of gypsy moths and forest tent caterpillars

Individual quaking aspen trees vary greatly in foliar chemistry and susceptibility to defoliation by gypsy moths and forest tent caterpillars. To relate performance of these insects to differences in foliar chemistry, we reared larvac from egg hatch to pupation on leaves from different aspen trees and analyzed leaf samples for water, nitrogen, total nonstructural carbohydrates, phenolic glycosides, and condensed tannins. Larval performance varied markedly among trees. Pupal weights of both species were strongly and inversely related to phenolic glycoside concentrations. In addition, gypsy moth performance was positively related to condensed tannin concentrations, whereas forest tent caterpillar pupal weights were positively associated with leaf nitrogen concentrations. A subsequent study with larvae fed aspen leaves supplemented with the phenolic glycoside tremulacin confirmed that the compound reduces larval performance. Larvae exhibited increased stadium durations and decreased relative growth rates and food conversion efficiencies as dietary levels of tremulacin increased. Differences in performance were more pronounced for gypsy moths than for forest tent caterpillars. These results suggest that intraspecific variation in defensive chemistry may strongly mediate interactions between aspen, gypsy moths and forest tent caterpillars in the Great Lakes region, and may account for differential defoliation of aspen by these two insect species.     

Anemotactic orientation of gypsy moth males and its modification by the attractant pheromone (+)-disparlure during walking

ABSTRACT. Tracks of dewinged gypsy moth males, Lymantria dispar L. (Lymantriidae), walking upwind in an airstream without pheromone consist of marked alternations between more or less straight upwind segments, partly with an arcadic structure, and twisted segments. This apparently complicated behaviour can, however, simply be explained by a superposition of noise and two turning commands: an upwind turning tendency, derived from the anemoreceptive system, which represents an average of the moth's angular positions over a period of time; and an internal turning tendency which consists of strong but brief bursts. These bursts are produced intermittently; successive bursts do not necessarily alternate polarity. Amputation of one antenna increases the probability of bursts towards the amputated side; therefore a separate burst source is postulated for each antenna. In the presence of the attractant pheromone (+)-disparlure, the anemotactic signal is weighted higher; twisted segments are, therefore, less pronounced.
There is a chemotropotactical component involved in the male's orientation. The tropotactical signal, dependent on the difference of odour concentration perceived by the left and right antenna, competes with the upwind turning tendency.     

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.     

Discrimination and production of disparlure enantiomers by the gypsy moth and the nun moth

ABSTRACT. Two odour receptor cells were physiologically identified within male antennal hair sensillae of the gypsy moth, Lymantria dispar L, and the nun moth, L. monacha L. In the gypsy moth, one cell responded to (+)-disparlure, while a neighbouring cell responded to (-)-disparlure. In the nun moth both cells responded to (+)-disparlure. The lack of sensitivity to (-)-disparlure in the nun moth was corroborated by electroantennogram (EAG) recordings, which indicated no affinity to this enantiomer. Single cell responses of male gypsy moth to different concentrations of the synthetic enantiomers of disparlure were then compared to responses elicited by hexane extracts of female glands of both species. The gypsy moth's extracts stimulated almost exclusively the receptor cell specialized for (+)-disparlure, while both cells were simultaneously stimulated by the extracts of the nun moths. From the response characteristic of the cells it is estimated that pheromone production of the nun moth is about 10% (+) and 90% (-)-disparlure, and that of the gypsy moth is almost 100% (+)-disparlure. Stimulation of the antenna of each species by female gland extracts of both species did not indicate the presence of receptors for other hexane elutable pheromone components in either species.     

Temporal events of gypsy moth vitellogenesis and ovarian development

Abstract The vitellogenic period of gypsy moth ovarian development starts on day 3 of the pupal stage and continues through adulthood. During this period, rapid increases occur in follicle size, protein content, and wet weight of the ovary. Patency is observed on day 3 of the pupal stage.
Pre-vitellogenic follicles are formed in the last larval stadium. Newly formed follicles detach from the germarium on day 4, and increase rapidly to 140 per ovariole at the end of the last larval stadium. The pre-vitellogenic follicles are uniformly around 50 um in diameter. No vitellogenin is incorporated into the oocytes until the pupal stage.
Polyacrylamide gel electrophosesis (PAGE) in the presence of sodium dodecylsulphate (SDS) analysis of male and female haemolymph samples and vitellogenic ovaries demonstrates the presence of two female-specific subunits of vitellogenin of 180 kD and 160 kD. These proteins are detected only in haemolymph and ovarian extracts of vitellogenic females. The molecular weight of the native protein determined by size exclusion chromatography is approximately 400–420 kD.
A highly sensitive double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed to monitor the temporal changes in vitellogenin titre in haemolymph. Vitellogenin production starts on day 2 of the last larval stadium, reaching a maximum level by day 6 of the last larval stadium, and decreasing in the late pupal stage as vitellogenin was internalized into the oocytes. This is the first report of vitellogenin production occurring in the larval stage of a holometabolous insect. The fact that vitellogenin production and uptake occur during different stages of development in the gypsy moth, opens up some interesting questions concerning the underlying regulatory mechanisms controlling each process.     

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.     

Risk assessment of the gypsy moth, <Emphasis Type="Italic">Lymantria dispar</Emphasis> (L), in New Zealand based on phenology modelling

The gypsy moth is a global pest that has not yet established in New Zealand despite individual moths having been discovered near ports. A climate-driven phenology model previously used in North America was applied to New Zealand. Weather and elevation data were used as inputs to predict where sustainable populations could potentially exist and predict the timing of hatch and oviposition in different regions. Results for New Zealand were compared with those in the Canadian Maritimes (New Brunswick, Nova Scotia, and Prince Edward Island) where the gypsy moth has long been established. Model results agree with the current distribution of the gypsy moth in the Canadian Maritimes and predict that the majority of New Zealand’s North Island and the northern coastal regions of the South Island have a suitable climate to allow stable seasonality of the gypsy moth. New Zealand’s climate appears more forgiving than that of the Canadian Maritimes, as the model predicts a wider range of oviposition dates leading to stable seasonality. Furthermore, we investigated the effect of climate change on the predicted potential distribution for New Zealand. Climate change scenarios show an increase in probability of establishment throughout New Zealand, most noticeably in the South Island.     

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.     

Protein intake by gypsy moth larvae on homogeneous and heterogeneous diets

Abstract. Food selection behaviour, food utilization efficiency and growth performance of a generalist insect, the gypsy moth ( Lymantria dispar (L.), Lepidoptera: Lymantriidae), were examined with respect to variation in food nitrogen concentration. The results suggest that gypsy moth do not suffer physiologically and in fact may benefit from intraplant variation by selective feeding. When provided with diet cubes containing identical nitrogen concentrations, control larvae tended to consume food from a single cube. This behaviour contrasted with that of larvae provided cubes differing in nitrogen concentration. These larvae tended to consume more from the high nitrogen cube, but allocated feeding more evenly among diet cubes than did control larvae. Overall, larvae mixed foods so as to obtain a mean concentration of 2.9-3.2% nitrogen, a concentration assumed to approximate the 'intake target'. Larvae confined to single nitrogen concentrations mitigated the impact of imbalanced diets on body composition via both pre-ingestive and post-ingestive compensation. When confined to a specific nitrogen concentration, larvae adjusted their intake to the point of best compromise. In this case, this was the geometrically closest point to the estimated intake target. Larvae with a choice of foods that deviated more than ±1% from each other in nitrogen concentration grew as well as or better than larvae without a choice but given identical mean nitrogen concentrations. These results demonstrate that selectivity and nitrogen consumption by gypsy moth larvae are altered according to the particular choices available. Insects may benefit from intraplant variation in food quality because such variation provides the opportunity to choose foods and mix them in ways that permit close matching with the intake target. Variation may be particularly important to insects which must offset changing nutritional demands.     

The effects of host plant defoliation and fertilizer application on larval growth and oviposition behaviour in cinnabar moth

Summary Defoliated ragwort plants produced regrowth foliage that was higher in alkaloid, but lower in amino acid concentrations than primary foliage. Total N was not affected. 2) Plants fertilized with nitrogen (as ammonium sulphate) had lower amino acid concentrations than unfertilized control plants, slightly increased alkaloid levels but similar total N concentrations. 3) Ovipositing females laid eggs upon plants with equal probability for controls, regrowth and fertilized foliage (one rosette in 5 received an egg batch). However, the probability of receiving eggs was significantly lower on the primary leaves of cut-back plants that had had their lower leaves removed a few days before egg laying (only one rosette in 13 was selected). 4) Egg batch size was higher on fertilized control foliage than on other treatments. 5) Larvae attained greater final weights when fed a diet of regrowth foliage, despite the higher levels of alkaloid they contained. Larval development rate was not affected by experimental treatment of the foliage. 6) Larval growth was lowest on the leaves of fertilized plants. This was associated with significant reductions in the concentrations of three amino acids (methionine down 29%, tyrosine 33% and lysine 25%).     

Improved insect performance from host-plant defoliation: winter moth on oak and apple

ABSTRACT. 1. We test the hypothesis that defoliation has short-term and long-term negative effects on performance of winter moth Operophtera brumata L. on two species of food plants: Garry oak ( Quercus garryana Dougl.) and apple ( Malus domesticus L.).
2. Pupal weight (and hence potential fecundity of females) decreased with increased defoliation in the current year on both tree species.
3. Pupal weight increased, however, with increased level of defoliation in the previous year.
4. Increased weight of pupae from larvae which had fed on previously defoliated trees may buffer population decline by increasing fecundity of moths in the next generation.     

Brush border membrane aminopeptidase-n in the midgut of the gypsy moth serves as the receptor for the CryIA(c) δ-endotoxin of Bacillus thuringiensis

Aminopeptidase-N (AP-N) was purified from gypsy moth (Lymantria dispar, L.) brush border membrane vesicles (BBMV) proteins by mono-Q chromatography and Superdex-75 gel filtration in the presence of the zwitterionic detergent, CHAPS, using FPLC. The purified AP-N, identified by its enzymatic activity, had an apparent size of 100 kDa, and was identified as the unique Bacillus thuringiensis insecticidal toxin, CryIA(c), binding protein. AP-N clearly displayed strong binding to CryIA(c), exhibiting little or no binding to CryIA(a) or CryIA(b), and showing no binding for the coleopteran-specific toxin, CryIIIA. Protein blots of the BBMV proteins probed with biotin-labeled and ¹²⁵I-labeled insecticidal proteins revealed that CryIAc binds only to 120 kDa protein which is a slightly larger size in comparison to purified AP-N. Antibodies raised against the gypsy moth AP-N demonstrated that the purified AP-N and the 120 kDa CryIA(c) binding protein of total BBMV proteins are antigenically identical.     

Physiological basis for sterilizing effects of constant light in Lymantria dispar

ABSTRACT Females of gypsy moth, Lymantria dispar L. (Lepidoptera: Lymantriidae), mated to males kept in constant light (LL) as pharate adults fail to oviposit. In males, a rhythm of sperm release from the testis that occurs in light-dark (LD) cycles is abolished in LL, and the total amount of sperm released from the testis is approximately half of that of LD males. Moreover, any sperm that may be released from the testis of LL males tend to remain in the vasa deferentia instead of moving into the duplex as in LD males. Consequently, in LL very few sperm bundles are transferred to the bursa copulatrix during mating; furthermore, these bundles fail to disperse into spermatozoa and sperm do not reach the spermatheca. The presence of a spermatheca filled with sperm must play an important role in controlling oviposition because their removal from mated females prevents egg-laying. Our results indicate that the rhythm of sperm release from the testis is essential for the ability of sperm to migrate in male and female reproductive tracts. The rhythms may help to synchronize final stages of sperm development with the activity of phagocytic and secretory cells lining the reproductive tract.