Intrastadial developmental resistance of third instar gypsy moths (Lymantria dispar L.) to L. dispar nucleopolyhedrovirus

Gypsy moth larvae become increasingly resistant to lethal infection by the Lymantria dispar M nucleopolyhedrovirus (LdMNPV) as they age within the fourth instar. Newly molted larvae are most sensitive to infection, mid-instars are least sensitive, and late-instars display intermediate sensitivity. This resistance occurs whether the virus is delivered orally or intrahemocoelically. The present study reveals a nearly identical pattern of resistance in third instar larvae. An LD₄₈ dose of polyhedra for newly molted third instars produced 18%, 10%, 8%, 25%, and 24% mortalities in larvae to which virus was orally administered at 12, 24, 48, 72, and 96 hours post-molt (hpm), respectively, which is a 6-fold reduction in mortality between newly molted larvae and mid-instars. An LD₄₄ dose of budded virus for newly molted third instars produced 33%, 23%, 17%, 31%, and 31% mortalities when injected into larvae that were 12, 24, 48, 72, and 96 hpm, respectively, which is a 2.6-fold reduction in mortality between newly molted larvae and mid-instars, indicating that approximately half of this resistance is midgut-based and half is systemically based. Doubling the viral dose did not overcome developmental resistance whether the virus was delivered orally or intrahemocoelically. In addition, time to death was significantly affected by the time post-molt at which the insect was inoculated with the virus. We suggest that intrastadial developmental resistance may affect both the ecology and management of the gypsy moth.     

Factors affecting immigration of adults: experimental and theoretical observations with rodents

We examined immigration in populations of Peromyscus leucopus (white-footed mice) by 1) monitoring natural immigration at 10 sites, 2) introducing experimental immigrants into eight populations, and 3) constructing qualitative models of immigration. Density of natural immigrants covaried positively with resident density, and successful assimilation was lower at low resident and immigrant densities. Females and males did not differ in their chance of achieving residency. Year, sex, and number introduced were significant predictors of assimilation by individuals. Experimentally introduced individuals had no effect on densities of those mice resident before the introductions. Results obtained from studying natural immigration differed from those obtained from studying experimental immigration. Signed digraphs and time averaging were used to model the consequences of different resident-immigrant relationships and to suggest how different sources of variation may have affected assimilation and led to differences in natural and experimental immigration. Resident and immigrant densities could have been positively correlated even if residents actively inhibited immigration. Variation in immigrant density and survival rather than resident territorial activity apparently determined patterns of assimilation.     

Estrous phase alters social behavior in a polygynous but not a monogamous Peromyscus species

The social organization of rodent species determines behavioral patterns for both affiliative and agonistic encounters. The neuropeptide oxytocin has been implicated in the mediation of social behavior; however, variability in both neuropeptide expression and social behavior within a single species indicates an additional mediating factor. The purpose of the present comparative study was to investigate social behaviors in naïve mixed-sex pairs of monogamous Peromyscus californicus and polygynous Peromyscus leucopus. We identified substantial inter- and intra-specific variability in the expression of affiliative and agonistic behaviors. Although all P. californicus tested engaged in frequent and prolonged intervals of social contact and rarely engaged in aggressive behaviors, P. leucopus exhibited significant variability in both measures of social behaviors. The naturally occurring differences in social behavior displayed by P. leucopus vary across the estrous cycle, and correspond to hypothalamic oxytocin, as well as circulating oxytocin and glucocorticoid concentrations. These results provide evidence for a rhythm in social behavior across the estrous cycle in polygynous, but not monogamous, Peromyscus species.     

Induced plant defenses breached? Phytochemical induction protects an herbivore from disease

Although wound-induced responses in plants are widespread, neither the ecological nor the evolutionary significance of phytochemical induction is clear. Several studies have shown, for example, that induced responses can act against both plant pathogens and herbivores simultaneously. We present the first evidence that phytochemical induction can inhibit a pathogen of the herbivore responsible for the defoliation. In 1990, we generated leaf damage by enclosing gypsy moth larvae on branches of red oak trees. We then inoculated a second cohort of larvae with a nuclear polyhedrosis virus (LdNPV) on foliage from the damaged branches. Larvae were less susceptible to virus consumed on foliage from branches with increasing levels of defoliation, and with higher concentrations of gallotannin. Defoliation itself was not related to any of our chemistry measures. Field sampling supported the results of our experiments: death from virus among feral larvae collected from unmanipulated trees was also negatively correlated with defoliation. In 1991, defoliation and gallotannin were again found to inhibit the virus. In addition, gallotannin concentrations were found to be positively correlated with defoliation the previous year. Compared with previous results that demonstrated a delecterious effect of induction on gypsy moth pupal weight and fecundity, the inhibition of the virus should confer an advantage to the gypsy moth. Since leaf damage levels increase as gypsy moth density increases, and since leaf damage inhibits the gypsy moth virus, there is the potential for positive feedback in the system. If phytochemical induction in red oak can inhibit an animal pathogen such as LdNPV, it suggests to us that induction in red oak is a generalized response to tissue damage rather than an adaptive defense against herbivores.     

Larvicidal and antifeedant activity of some plant-derived compounds to Lymantria dispar L. (Lepidoptera: Limantriidae)

Ethanol solutions of essential oil of Ocimum basilicum and its main component, linalool (both isomer forms), all in three concentrations, as well as botanical standard Bioneem (0.5%), were tested for their toxicity and antifeedant activity against the second instar gypsy moth larvae in the laboratory bioassay. The essential oil of O. basilicum was subjected to gas chromatography analysis, and totally 37 compounds were detected, of which linalool was predominantly present. All tested solutions showed low to moderate larvicidal effect in both residual toxicity test and in chronic larval mortality bioassay. Chronic mortality tests showed that obtained mortality was a consequence of starving rather than ingestion of treated leaves. However, antifeedant index achieved by application of tested solutions in feeding choice assay was remarkable. Foliar application of all tested compounds deterred feeding by L2 in the same percent as Bioneem. Antifeedant index was relatively high at all tested treatments (85-94%); moreover, the larval desensitization to repelling volatiles has not occurred after five days of observation. Low toxic and high antifeedant properties make these plant-derived compounds suitable for incorporation in integrated pest management programs, especially in urban environments.     

Interspecific competition between the braconid endoparasitoids Glyptapanteles porthetriae and Glyptapanteles liparidis in Lymantria dispar larvae

The effect of interspecific competition between the solitary endoparasitoid Glyptapanteles porthetriae Muesebeck (Hymenoptera: Braconidae) and the gregarious Glyptapanteles liparidis Bouché (Hymenoptera: Braconidae), was investigated in larvae of Lymantria dispar L. (Lepidoptera: Lymantriidae). Host larvae were parasitized by both wasp species simultaneously in premolt to the 2^nd or the 3^rd host instar or in an additional approach with a 4-day delay in parasitization by the second wasp species. Host acceptance experiments revealed that both wasp species do not discriminate between unparasitized host larvae and larvae parasitized previously by the same or the other species. In more than 90% female wasps parasitized the larva they encountered first. During the period of endoparasitic development, larvae of the competing parasitoid species never attacked the egg stage of the other species. When host larvae were parasitized simultaneously by both wasp species, the rate of successful development of both species depended on the age of the host larva at the time of its parasitization; G. liparidis emerged successfully from 44% of host larvae parasitized during the premolt to 2^nd instar, G. porthetriae from 28%, and in 20% of the hosts both parasitoid species were able to develop in one gypsy moth larva. However, when host larvae were parasitized simultaneously during premolt to the 3^rd instar, G. liparidis was successful in 90% of the hosts, compared to 8% from which only G. porthetriae emerged. In the experiments with delayed oviposition, generally the species that oviposited first succeeded in completing its larval development. Larvae of the species ovipositing with four days delay were frequently attacked and killed by larvae of the first parasitizing species or suffered reduced growth. As the secondary parasitoid species, G. porthetriae-larvae were never able to complete their development, whereas G. liparidis developed successfully in at least 12,5% of the multiparasitized host larvae. Thus, multiparasitism of gypsy moth larvae by both Glyptapanteles species corresponds to the contest type; however, G. porthetriae is only able to develop successfully as the primary parasitoid of young host larvae.     

Localization of Bacillus thuringiensis Cry1A toxin-binding molecules in gypsy moth larval gut sections using fluorescence microscopy

The microbial insecticide Bacillus thuringiensis (Bt) produces Cry toxins, proteins that bind to the brush border membranes of gut epithelial cells of insects that ingest it, disrupting the integrity of the membranes, and leading to cell lysis and insect death. In gypsy moth, Lymantria dispar, two toxin-binding molecules for the Cry1A class of Bt toxins have been identified: an aminopeptidase N (APN-1) and a 270 kDa anionic glycoconjugate (BTR-270). Studies have shown that APN-1 has a relatively weak affinity and a very narrow specificity to Cry1Ac, the only Cry1A toxin that it binds. In contrast, BTR-270 binds all toxins that are active against L. dispar larvae, and the affinities for these toxins to BTR-270 correlate positively with their respective toxicities. In this study, an immunohistochemical approach was coupled with fluorescence microscopy to localize APN-1 and BTR-270 in paraffin embedded midgut sections of L. dispar larvae. The distribution of cadherin and alkaline phosphatase in the gut tissue was also examined. A strong reaction indicative of polyanionic material was detected with alcian blue staining over the entire epithelial brush border, suggesting the presence of acidic glycoconjugates in the microvillar matrix. The Cry1A toxin-binding sites were confined to the apical surface of the gut epithelial cells with intense labeling of the apical tips of the microvilli. APN-1, BTR-270, and alkaline phosphatase were found to be present exclusively along the brush border microvilli along the entire gut epithelium. In contrast, cadherin, detected only in older gypsy moth larvae, was present both in the apical brush border and in the basement membrane anchoring the midgut epithelial cells. The topographical relationship between the Bt Cry toxin-binding molecules BTR-270 and APN-1 and the Cry1A toxin-binding sites that were confined to the apical brush border of the midgut cells is consistent with findings implicating their involvement in the mechanism of the action of Bt Cry toxins.     

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.     

Positive effects of testosterone and immunochallenge on energy allocation to reproductive organs

A number of studies have suggested the incompatibility of simultaneous increases in immune and reproductive functions. Other research has indicated that immune responses may be modulated depending on the relative benefits of increased survival and prospects for current and future reproduction. We tested the hypothesis that energy allocation to reproductive and other organ systems is not affected by testosterone level and energy expenditure on immune functions. Adult male white-footed mice (Peromyscus leucopus) with or without elevated testosterone levels and with or without immunochallenges were tested. Testosterone treatment was associated with reduced humoral immune response indicating immunosuppressive effects, reduced masses of gastrointestinal organs, reduced corticosterone level, increased kidney and seminal vesicle masses, and increased hematocrit. Immunochallenge was associated with increased resting metabolic rate and testes and seminal vesicle masses. Reproductive organ masses were greatest in immunochallenged mice with exogenous testosterone. Simultaneous increases in energy allocation to immune and reproductive structures may be an adaptive response that would enhance survival and current prospects for reproduction.     

Survey ofLymantria obfuscata and its natural enemies in India

The status ofLymantria obfuscata Walker in India is discussed. Notes on its taxonomy, distribution, life-history and economic importance are given for comparison with those of the gypsy moth,Lymantria dispar (L.). Parasites ofL. obfuscata in India are reviewed. A survey in 1961–66 revealed an additional 50 species of natural enemies ofL. obfuscata in India. The incidence and biology of some of the important ones are presented. A comparison is made between these parasites and those ofL. dispar. Some of the natural enemies ofL. obfuscata already attackL. dispar in America. Others are closely related to the parasites of the gypsy moth. The scope for utilisation of parasites ofL. obfuscata in other countries is discussed. Possible use of parasites ofL. dispar from the U.S.A. or elsewhere againstL. obfuscata in India is suggested. Research financed in part by the U.S.D.A. under grant PL-480.     

Development of gypsy moth larvae feeding on red maple saplings at elevated CO<Subscript>2</Subscript> and temperature

Predicted increases in atmospheric CO₂ and global mean temperature may alter important plant-insect associations due to the direct effects of temperature on insect development and the indirect effects of elevated temperature and CO₂ enrichment on phytochemicals important for insect success. We investigated the effects of CO₂ and temperature on the interaction between gypsy moth (Lymantria dispar L.) larvae and red maple (Acer rubrum L.) saplings by bagging first instar larvae within open-top chambers at four CO₂/temperature treatments: (1) ambient temperature, ambient CO₂, (2) ambient temperature, elevated CO₂ (+300 l l^-1 CO₂), (3) elevated temperature (+3.5°C), ambient CO₂, and (4) elevated temperature, elevated CO₂. Larvae were reared to pupation and leaf samples taken biweekly to determine levels of total N, water, non-structural carbohydrates, and an estimate of defensive phenolic compounds in three age classes of foliage: (1) immature, (2) mid-mature and (3) mature. Elevated growth temperature marginally reduced (P <0.1) leaf N and significantly reduced (P <0.05) leaf water across CO₂ treatments in mature leaves, whereas leaves grown at elevated CO₂ concentration had a significant decrease in leaf N and a significant increase in the ratio of starch:N and total non-structural carbohydrates:N. Leaf N and water decreased and starch:N and total non-structural carbohydrates:N ratios increased as leaves aged. Phenolics were unaffected by CO₂ or temperature treatment. There were no interactive effects of CO₂ and temperature on any phytochemical measure. Gypsy moth larvae reached pupation earlier at the elevated temperature (female =8 days, P <0.07; male =7.5 days, P <0.03), whereas mortality and pupal fresh weight of insects were unrelated to either CO₂, temperature or their interaction. Our data show that CO₂ or temperature-induced alterations in leaf constituents had no effect on insect performance; instead, the long-term exposure to a 3.5°C increase in temperature shortened insect development but had no effect on pupal weight. It appears that in some tree-herbivorous insect systems the direct effects of an increased global mean temperature may have greater consequences for altering plant-insect interactions than the indirect effects of an increased temperature or CO₂ concentration on leaf constituents.     

Use of learned visual cues during habitat location by Brachymeria intermedia

Like many other parasitoids, Brachymeria intermedia (Hymenoptera: Chalcididae) uses olfactory cues to find its host, but in addition it appears to use learned visual cues to focus its searching efforts at the macrohabitat level. In laboratory experiments, females were held in cages where Lymantria dispar pupae were hidden either in a vertical, tree-like structure or on the floor. After four days females had learned to search for hosts in the structure in which they previously had found pupae. Such training was reversible. During similar tests in a semi-natural situation, in which pupae were hung from a tree trunk or were hidden under leaf litter, females also tailored their subsequent searching to favour the macrohabitat where hosts previously had been found. This parasitoid most likely uses a combination of visual and olfactory cues during host searching.     

Daily rhythm in myogenic contractions of vas deferens associated with sperm release cycle in a moth

In the gypsy moth, Lymantria dispar, release of sperm bundles from the testis into the upper vas deferens (UVD) and subsequent transfer of sperm bundles into the seminal vesicles (SV) occurs in a daily rhythm. The UVD undergoes different types of contractions despite the fact that its musculature appears to receive no innervation. Patterns of the UVD movements were recorded throughout the daily sperm release and transfer cycle. In males kept in light-dark cycles, transfer of sperm from the UVD to the SV was accompanied by a characteristic pattern of UVD contractions of high frequency and amplitude. In males kept in constant light, which fail to transfer sperm, this contraction pattern was absent. It is concluded that the vas deferens muscles undergo daily changes in contraction pattern in phase with the light-dark cycle. The increased muscular contractions appear to be a causal factor in the gated sperm transfer from the UVD to the SV.Abbreviations LD light-dark - LL constant light - SV seminal vesicle - UVD upper vas deferens     

Temporal consistency in the spatial pattern of seed predation across a forest – old field edge

Seed predation is an important factor in determining the rate of tree establishment in abandoned agricultural land. Edges, through altered habitat use by small mammals, may influence the spatial pattern of forest regeneration in these successional sites. To determine the spatial pattern of seed predation across a forest-old field edge, we used a grid that began 30 m inside the forest and extended 60 m into the old field. Seed stations were placed at regular 10-m intervals and were monitored for removal of Acer rubrum seed for 50 d. This design was repeated over four years (1995–1998). Small mammal trapping was conducted in the final year of the study to determine the spatial pattern of seed predators within the site. Removal rates varied among the four years of the study with years of high and low removal rates. However, the spatial pattern of seed removal rate was similar in all years. Final survival ranged from 0.7–15.5% of seeds, with lower final survival in years with faster rates of seed removal (1996 and 1998). Seed removal rates and rates of discovery were greatest at the forest edge and decreased with distance into the old field. The number of seeds surviving to the end of the experiment also varied across the edge gradient, with highest survival at greater distances into the old field in low predation years. Seed removal rate covaried with spatial pattern of Peromyscus leucopus captures within the site. Seed removal and discovery was also higher under the exotic shrub Rosa multiflora, which may have provided cover for foraging seed predators. These indirect effects of edges on plant communities can potentially alter the rate and spatial pattern of tree invasion into disturbed lands and illustrate the importance of understanding plant-animal interactions in the context of habitat fragmentation.     

Retrotransposon Mys was active during evolution of the Peromyscus leucopus-maniculatus complex

Mys is a retrovirus-like transposable element found throughout the genus Peromyscus. Several mys subfamilies identified on the basis of restriction site variation occur in more than one species. The distribution of these subfamilies is consistent with the accepted species phylogeny, suggesting that mys was present in the ancestor of Peromyscus and has been active through much of the evolution of this genus. Quantitative Southern blot analysis was used to examine the variability of subfamilies in P. leucopus and maniculatus. We found that subfamilies with phylogenetically narrow distributions were more variable in copy number both within and between species than subfamilies with a broader distribution. Taken together, our data suggest that mys has undergone multiple rounds of transposition since the peromyscine radiation, and that five subfamilies have been amplified during the evolution of the leucopus-maniculatus species complex. Correspondence to: H.A. Wichman