Gypsy Moth Caterpillars Research Articles

Metabarcoding of canopy arthropods reveals negative impacts of forestry insecticides on community structure across multiple taxa

Abstract Insecticides used to combat outbreaks of forest defoliators can adversely affect non‐target arthropods. Forestry insecticides typically suppress Lepidoptera larvae which are the cornerstone of the canopy community of deciduous oak forests. The abrupt removal of this dominant component of the food web could have far‐reaching implications for forest ecosystems, yet it is rarely investigated in practice owing to several methodological shortcomings. The taxonomic impediment and the biased nature of arthropod sampling techniques particularly impede the assessment of insecticide impacts on diverse communities. To tackle this issue, we propose an experimental approach combining sampling by pyrethrum knockdown and species determination via DNA metabarcoding, using community subsampling to derive estimates of species abundances. We applied this protocol to investigate the short‐term effects of the insecticides diflubenzuron (DFB) and Bacillus thuringiensis var. kurstaki (BTK) on canopy‐dwelling arthropod communities in German oak woodlands. Our approach allowed us to detect most of the diversity and integrate species abundances in our analyses. By classifying arthropod species into assemblages based on their expected sensitivity rather than coarse taxonomic groupings, we could unveil substantial effects of DFB across multiple taxa 5 weeks after application. Although strong effects on single species appear related to direct toxicity, substantial impacts of DFB on parasitoids and xylophagous beetles suggest that anti‐defoliator treatments can have previously unsuspected indirect effects on some components of forest arthropod communities. The impacts of BTK on community structure were consistent with that of DFB though much weaker. Synthesis and applications. Comparing diversity patterns in the arthropod communities of sprayed and unsprayed oak canopies, our results show that selective insecticides can alter species diversity in presumably non‐sensitive taxa. Even though the ecological significance of these impacts has yet to be assessed in an operational setting, their existence calls for increased regulatory scrutiny on indirect effects. As community approaches become more attainable with the rapid development of DNA metabarcoding, we suggest the inclusion of community‐level end points as regulatory requirements for the approval of forest‐use insecticides.

Чувствительность гусениц непарного шелкопряда (Lepidoptera: Erebidae: Lymantria dispar) с юга Хабаровского края к различным штаммам вируса ядерного полиэдроза

The article discusses data on the effect of the nuclear polyhedrosis virus (NPV) on gypsy moth caterpillars of the Far Eastern population. In particular, the article provides data on the dynamics of caterpillar mortality under laboratory conditions under different viral load of four NPV strains: Kyrgyz, Japanese, Krasnodar, and Siberian. The difference in the duration of the incubation period of a strain and infectious load has not been established. The maximum virulence was observed for the strain Kyrgyz (about 96% at the maximum titer). The duration of the mortal period differed for all tested strains. The indicator L50 was achieved for all infectious loads of the Kyrgyz and Krasnodar strains. The maximum daily mortality was detected at the maximum infectious loads of the Kyrgyz and Japanese strains (29.7% and 39.9%, respectively) on the second day of the mortal period.

Biological control of gypsy moth (Lymantria dispar) by the entomopathogenic fungus Entomophaga maimaiga in Bulgaria in 2021

In 2021, biological control programme against gipsy moth (Lymantria dispar) populations was carried out by introduction of the entomopathogenic fungus Entomophaga maimaiga on the territory of four State Forest Enterprises: Municipal Enterprise (ME) ‘Management of Municipal Forests, Agriculture and Forestry’, Nessebar; State Game Enterprises (SGE) Nessebar and Balchik; State Forestry (SF) Vidin. The pathogen was introduced during the period 15-26.03.2021 in 34 localities - five in ME Nessebar, eight in SGE Nessebar, ten in SGE Balchik and eleven in SF Vidin. The average number of gypsy moth population density in the locations of introduction was relatively high, ranging between 0.4-15.9 egg mass/tree in the area of ​​SGE Balchik and 11.9-65.0 egg mass/tree in the area of ​​ME Nessebar. The average mortality of young gypsy moth caterpillars (first-third instar) due to E. maimaiga varied between 2.6% (SGE Balchik) and 13.0% (SF Vidin), and of caterpillars in later fourth-sixth instar - between 20.7% (SF Vidin) and 52.4% (ME Nessebar). The overall mortality of the gipsy moth caterpillars due to E. maimaiga was lowest in the region of SGE Balchik (26.1%), followed by SF Vidin (33.7%), SGE Nessebar (48.5%) and ME Nessebar (55.9%). As a result of the introduction, gipsy moth severe outbreaks in the region of Nessebar was significantly suppressed. The high number of E. maimaiga resting spores persists in the surface layers of the soil in the other two areas (Vidin and Balchik) has the potential to suppress L. dispar attacks in next years.

Histologic lesions of experimental infection with Lymantria dispar multicapsid nucleopolyhedrovirus and Lymantria dispar cytoplasmic polyhedrosis virus in European gypsy moth caterpillars (Lymantria dispar dispar).

European gypsy moths (Lymantria dispar dispar) are an invasive species in North America, and are listed by the International Union for the Conservation of Nature as one of the 100 most destructive invasive species worldwide. They have several known viruses, some of which are used as biological control agents. However, there are no detailed descriptions of many entomopathogenic viral infections, including in European gypsy moths, using bright-field microscopy. In this study, 11 European gypsy moth caterpillars were evaluated histologically: 4 were experimentally infected with Lymantria dispar multicapsid nucleopolyhedrovirus (LdMNPV; Baculoviridae); 4 were experimentally infected with Lymantria dispar cytoplasmic polyhedrosis virus (LdCPV; Reoviridae); 3 control animals were uninfected. A complete tissue set was evaluated in all animals from all groups using bright-field microscopy, including epidermis, cuticle, striated muscle, tracheae, foregut, midgut, hindgut, Malpighian tubules, hemocytes, fat body, and nervous system. LdMNPV-infected caterpillars had marked karyomegaly and intranuclear viral inclusions in cells of the epidermis, tracheae, fat body, and hemocytes. LdMNPV-infected caterpillars also had hyperplasia and hypertrophy of epidermal and tracheal epithelial cells. LdCPV-infected caterpillars had numerous granular eosinophilic intracytoplasmic viral inclusions in midgut epithelial cells. Both LdMNPV-infected and LdCPV-infected caterpillars had atrophy of fat body adipocytes; this change was more pronounced in LdCPV-infected caterpillars. This work provides the first detailed descriptions of these viral infections in European gypsy moth caterpillars using bright-field light microscopy and provides images of normal histology from control caterpillars.

Defoliation severity is positively related to soil solution nitrogen availability and negatively related to soil nitrogen concentrations following a multi-year invasive insect irruption.

Understanding connections between ecosystem nitrogen (N) cycling and invasive insect defoliation could facilitate the prediction of disturbance impacts across a range of spatial scales. In this study we investigated relationships between ecosystem N cycling and tree defoliation during a recent 2015–18 irruption of invasive gypsy moth caterpillars (Lymantria dispar), which can cause tree stress and sometimes mortality following multiple years of defoliation. Nitrogen is a critical nutrient that limits the growth of caterpillars and plants in temperate forests. In this study, we assessed the associations among N concentrations, soil solution N availability and defoliation intensity by L. dispar at the scale of individual trees and forest plots. We measured leaf and soil N concentrations and soil solution inorganic N availability among individual red oak trees (Quercus rubra) in Amherst, MA and across a network of forest plots in Central Massachusetts. We combined these field data with estimated defoliation severity derived from Landsat imagery to assess relationships between plot-scale defoliation and ecosystem N cycling. We found that trees in soil with lower N concentrations experienced more herbivory than trees in soil with higher N concentrations. Additionally, forest plots with lower N soil were correlated with more severe L. dispar defoliation, which matched the tree-level relationship. The amount of inorganic N in soil solution was strongly positively correlated with defoliation intensity and the number of sequential years of defoliation. These results suggested that higher ecosystem N pools might promote the resistance of oak trees to L. dispar defoliation and that defoliation severity across multiple years is associated with a linear increase in soil solution inorganic N.

Short term effects of Bacillus thuringiensis and diflubenzuron aerial applications on non-target arthropods in a Mediterranean Forest

Several products are widely used to control gypsy moth (Lymantria dispar) populations. The main purpose of the present study is to evaluate the aerial application effects of B. thuringiensis and diflubenzuron on non-target arthropods in a Mediterranean holm-oak forest. The density of gypsy moth population at the time of application of treatments was the same in the three forests and the defoliation levels in the three holm-oak forests were between level 0 (no apparent defoliation) and level 1 (small defoliations of new leaf). Sampling litter, soil, ground-dwelling, and epiphytic arthropods showed that the holm-oak forests treated with diflubenzuron had a better quantitative faunal outcome than the Control, whereas the highest taxonomic diversity was found in the site treated with B. thuringiensis. These results could probably be explained by a temporary competitive decrease due to the huge number of gypsy moth caterpillars on plants, as well as the enormous accumulation of their excrements in the soil during a prolonged period of time.

Aboveground phytochemical responses to belowground herbivory in poplar trees and the consequence for leaf herbivore preference

Belowground (BG) herbivory can influence aboveground (AG) herbivore performance and food preference via changes in plant chemistry. Most evidence for this phenomenon derives from studies in herbaceous plants but studies in woody plants are scarce. Here we investigated whether and how BG herbivory on black poplar (Populus nigra) trees by Melolontha melolontha larvae influences the feeding preference of Lymantria dispar (gypsy moth) caterpillars. In a food choice assay, caterpillars preferred to feed on leaves from trees that had experienced attack by BG herbivores. Therefore, we investigated the effect of BG herbivory on the phytochemical composition of P. nigra trees alone and in combination with AG feeding by L. dispar caterpillars. BG herbivory did not increase systemic AG tree defences like volatile organic compounds, protease inhibitors and salicinoids. Jasmonates and salicylic acid were also not induced by BG herbivory in leaves but abscisic acid concentrations drastically increased together with proline and few other amino acids. Leaf coating experiments with amino acids suggest that proline might be responsible for the caterpillar feeding preference via presumptive phagostimulatory properties. This study shows that BG herbivory in poplar can modify the feeding preference of AG herbivores via phytochemical changes as a consequence of root-to-shoot signaling.

Comparative efficacy of gypsy moth (Lepidoptera: Erebidae) entomopathogens on transgenic blight-tolerant and wild-type American, Chinese, and hybrid chestnuts (Fagales: Fagaceae).

American chestnut (Castanea dentata [Marsh.] Borkh.) was once the dominant hardwood species in Eastern North America before an exotic fungal pathogen, Cryphonectria parasitica (Murrill) Barr, functionally eliminated it across its range. One promising approach toward restoring American chestnut to natural forests is development of blight-tolerant trees using genetic transformation. However, transformation and related processes can result in unexpected and unintended phenotypic changes, potentially altering ecological interactions. To assess unintended tritrophic impacts of transgenic American chestnut on plant-herbivore interactions, gypsy moth (Lymantria dispar L.) caterpillars were fed leaf disks excised from two transgenic events, Darling 54 and Darling 58, and four control American chestnut lines. Leaf disks were previously treated with an LD50 dose of either the species-specific Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) or the generalist pathogen Bacillus thuringiensis subsp. kurstaki (Btk). Mortality was quantified and compared to water blank controls. Tree genotype had a strong effect on the efficacies of both pathogens. Larval mortality from Btk-treated foliage from only one transgenic event, Darling 54, differed from its isogenic progenitor, Ellis 1, but was similar to an unrelated wild-type American chestnut control. LdMNPV efficacy was unaffected by genetic transformation. Results suggest that although genetic modification of trees may affect interactions with other nontarget organisms, this may be due to insertion effects, and variation among different genotypes (whether transgenic or wild-type) imparts a greater change in response than transgene presence.

Ambient temperature-mediated enzymic activities and intestinal microflora in Lymantria dispar larvae.

To understand how ambient temperature affect the gypsy moth larvae, and provide a theoretical basis for pest control in different environments. Fourth instar gypsy moth larvae were incubating for 3 hr at 15℃, 20℃, 25℃, 30℃, 35℃, and 40℃, respectively. Afterward, digestive and antioxidant enzyme activities, total antioxidant capacity, and intestinal microflora community were analyzed to reveal how the caterpillars respond to ambient temperature stress. Results showed that both digestive and antioxidant enzymes were regulated by the ambient temperature. The optimum incubation temperatures of protease, amylase, trehalase, and lipase in gypsy moth larvae were 30℃, 25℃, and 20℃, respectively. When the incubation temperature was deviated optimum temperatures, digestive enzyme activities would be downregulated depending on the extent of temperature stress. In addition, glutathione S-transferase, peroxidase, catalase, and polyphenol oxidase would be activated under a sufferable temperature stress, but superoxide dismutase and carboxylesterase (CarE) would be inhibited. In addition, results showed that the top two abundant phyla were Proteobacteria and Firmicutes. The phylum Firmicutes abundance was decreased and phylum Proteobacteria abundance was increased by ambient temperature stress. Moreover, it suggested that gypsy moth caterpillars at different ambient temperature mainly differed from each other by Escherichia-Shigella and Bifidobacterium in control, Acinetobacter in T15, and Lactobacillus in T40, respectively.

Divergent behavioural responses of gypsy moth (Lymantria dispar) caterpillars from three different subspecies to potential host trees

Almost all previous work on host-plant selection by insect herbivores has focused on adult behaviour; however, immature life stages can also play an active role in host discrimination. The important forest pest Lymantria dispar (gypsy moth) has three recognised subspecies: the European, Asian, and Japanese gypsy moth. Unlike the other two subspecies, the European subspecies is characterised by a loss of female flight ability, which might impose a selective pressure on larvae to actively engage in host-plant selection. We therefore explored the interactions of early-instar larvae from laboratory colonies of each subspecies with four potential hosts of differing quality: oak, beech, maple, and pine—measuring larval survival and performance, feeding preferences, responses to host-derived odour cues, and the propensity to disperse from hosts via ballooning. Compared to larvae from the Asian and Japanese subspecies, larvae from the (American-originated) European gypsy moth colony exhibited (i) significantly lower survival on the poorest quality host (pine), (ii) an ability to discriminate among hosts via olfactory cues; and (iii) higher propensity to disperse from sub-optimal hosts. These results are consistent with the hypothesis that larvae from flightless female European Gypsy moth subspecies play a more active role in host-plant selection.

Herbivore-induced volatile emission from old-growth black poplar trees under field conditions

Herbivory is well known to trigger increased emission of volatile organic compounds (VOCs) from plants, but we know little about the responses of mature trees. We measured the volatiles emitted by leaves of old-growth black poplar (Populus nigra) trees after experimental damage by gypsy moth (Lymantria dispar) caterpillars in a floodplain forest, and studied the effect of herbivory on the transcript abundance of two genes involved in the biosynthesis of VOCs, and the accumulation of defence phytohormones. Herbivory significantly increased volatile emission from the experimentally damaged foliage, but not from adjacent undamaged leaves in the damaged branches (i.e., no systemic response). Methylbutyraldoximes, 4,8-dimethyl-1,3,7-nonatriene (DMNT), (Z)-3-hexenol and (E)-β-ocimene, amongst other compounds, were found to be important in distinguishing the blend of herbivore-damaged vs. undamaged leaves. Herbivory also increased expression of PnTPS3 (described here for the first time) and PnCYP79D6-v4 genes at the damaged sites, these genes encode for an (E)-β-ocimene synthase and a P450 enzyme involved in aldoxime formation, respectively, demonstrating de novo biosynthesis of the volatiles produced. Herbivore-damaged leaves had significantly higher levels of jasmonic acid and its conjugate (−)-jasmonic acid-isoleucine. This study shows that mature trees in the field have a robust response to herbivory, producing induced volatiles at the damaged sites even after previous natural herbivory and under changing environmental conditions, however, further studies are needed to establish whether the observed absence of systemic responses is typical of mature poplar trees or if specific conditions are required for their induction.

Stream response to an extreme drought-induced defoliation event

We assessed stream ecosystem-level response to a drought-induced defoliation event by gypsy moth caterpillars (Lymantria dispar) with high-frequency water quality sensors. The defoliation event was compared to the prior year of data. Based on long-term records of precipitation and drought indices, the drought of 2015–2016 in Rhode Island, USA was an extreme climatic event that preceded and likely precipitated the defoliation from insect infestation. Canopy cover in the riparian area was reduced by over 50% increasing light availability which warmed the stream and stimulated autotrophic activity. Frass and leaf detritus contributed particulate carbon and organic nutrients to the stream. Based on locally calibrated s::can spectro::lyser data, nitrate concentration and flux did not significantly increase during defoliation while orthophosphate concentration and flux did significantly increase during part of the defoliation period. Lower mean daily dissolved oxygen (DO) levels and wider diel cycles of DO indicated higher biological activity during the defoliation event. Stream metabolism metrics were also significantly higher during defoliation and pointed to heterotrophic activity dominating in the stream. The increases in stream metabolism were low compared to other studies; in streams with higher nutrient levels (e.g., in agricultural or urban watersheds) the increase in light and temperature could have a stronger influence on stream metabolism. The in-stream metabolic processes and nutrient fluxes observed in response to the drought-driven defoliation event resulted from the long-term deployment of high-frequency water sensors. The proliferation of these water sensors now enable studies that assess ecosystem responses to stochastic, unusual disturbances.

Leaf rust infection reduces herbivore-induced volatile emission in black poplar and attracts a generalist herbivore.

Plants release complex volatile blends after separate attack by herbivores and pathogens, which play many roles in interactions with other organisms. Large perennials are often attacked by multiple enemies, but the effect of combined attacks on volatile emission is rarely studied, particularly in trees. We infested Populus nigra trees with a pathogen, the rust fungus Melampsora larici-populina, and Lymantria dispar caterpillars alone and in combination. We investigated poplar volatile emission and its regulation, as well as the behavior of the caterpillars towards volatiles from rust-infected and uninfected trees. Both the rust fungus and the caterpillars alone induced volatile emission from poplar trees. However, the herbivore-induced volatile emission was significantly reduced when trees were under combined attack by the herbivore and the fungus. Herbivory induced terpene synthase transcripts as well as jasmonate concentrations, but these increases were suppressed when the tree was additionally infected with rust. Caterpillars preferred volatiles from rust-infected over uninfected trees. Our results suggest a defense hormone crosstalk upon combined herbivore-pathogen attack in poplar trees which results in lowered emission of herbivore-induced volatiles. This influences the preference of herbivores, and might have other far-reaching consequences for the insect and pathogen communities in natural poplar forests.

Complete Genome Sequence of Lymantria dispar multiple nucleopolyhedrovirus Isolated in Southwestern Poland.

We report here the complete genomic sequence of Polish alphabaculovirus isolated from dead gypsy moth caterpillars. Its genome structure and G+C content indicate that it is a Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) strain which possesses the shortest sequence among eight other (similar to reference sequence AF081810) LdMNPV sequences available in GenBank; it is 159,729 bp long.

Metabolism of poplar salicinoids by the generalist herbivore Lymantria dispar (Lepidoptera)

The survival of insect herbivores on chemically defended plants may often depend on their ability to metabolize these defense compounds. However, only little knowledge is available on how insects actually process most plant defense compounds. We investigated the metabolism of salicinoids, a major group of phenolic glycosides in poplar and willow species, by a generalist herbivore, the gypsy moth (Lymantria dispar). Seven salicinoid metabolites identified in gypsy moth caterpillar feces were mostly conjugates with glucose, cysteine or glycine. Two of the glucosides were phosphorylated, a feature not previously reported for insect metabolites of plant defense compounds. The origins of these metabolites were traced to specific moieties of three major poplar salicinoids ingested, salicin, salicortin and tremulacin. Based on the observed metabolite patterns we were able to deduce the initial steps of salicinoid breakdown in L. dispar guts, which involves cleavage of ester bonds. The conjugated molecules were effectively eliminated within 24 h after ingestion. Some of the initial breakdown products (salicin and catechol) demonstrated negative effects on insect growth and survival in bioassays on artificial diets. Gypsy moth caterpillars with prior feeding experience on salicinoid-containing poplar foliage converted salicinoids to the identified metabolites more efficiently than caterpillars pre-fed an artificial diet. The majority of the metabolites we identified were also produced by other common poplar-feeding insects. The conversion of plant defenses like salicinoids to a variety of water-soluble sugar, phosphate and amino acid conjugates and their subsequent excretion fits the general detoxification strategy found in insect herbivores and other animals.

Original Article. Topical treatment of LdMNPV-infected gypsy moth caterpillars with 18 nucleotides long antisense fragment from LdMNPV IAP3 gene triggers higher levels of apoptosis in infected cells and mortality of the pest

Abstract The high efficiency of baculovirus infection is partially explained by the ability of the virus to suppress host defense machinery connected with the apoptosis pathway. Members of the baculovirus gene family, inhibitors of apoptosis (vIAPs), have been shown to inhibit apoptosis in baculovirus-infected cells. Here we showed that treatment of the LdMNPV-infected 1st instar gypsy moth (Lymantria dispar) caterpillars with sense (oligoBIR) and antisense (oligoRING) DNA oligonucleotides from the LdMNPV IAP3 gene induced elevated mortality of the insects. Apoptotic DNA ladder assay showed that the leading role in this phenomenon is played by the antisense oligoRING fragment of the vIAP3 gene. These results imply that the application of both antisense DNA oligonucleotides from vIAP genes and baculovirus preparations (one following the other) may be a potential method for plant protection against insect pests.

An alphabaculovirus isolated from dead Lymantria dispar larvae shows high genetic similarity to baculovirus previously isolated from Lymantria monacha – An example of adaptation to a new host

A new isolate of baculovirus, Lymantria dispar multiple nucleopolyhedrovirus-BNP (LdMNPV-BNP), was found in dead gypsy moth (L. dispar) caterpillars collected in the Biebrzanski National Park in Poland. Here, we examined its biological activity, structure, genetic content and phylogeny. Multiple nucleocapsids of LdMNPV-BNP are enveloped together in 2–26 virions embedded in occluded bodies (OBs) very similar to the OBs previously described in viruses infecting Lymantriinae. This isolate kills pest larvae in a relatively short time (LT50 of approximately 9days for a dose of 2×107OBs/ml), highlighting the possibility for its use as a biopesticide. Next-generation sequencing of LdMNPV-BNP revealed gene content (e.g. DNA photolyase) that is not present in any LdMNPV isolate sequenced to date. The genome is 157,270 base pairs long and has a notably lower G+C content in comparison to other LdMNPVs (50.3% G+C content compared to an average of 57.4% among other LdMNPVs). According to our phylogenetic analysis based on 37 core genes, LdMNPV-BNP is a member of group II alphabaculoviruses, which are closely related to LdMNPV and LyxyMNPV (Lymantria xylina multiple nucleopolyhedrovirus). Molecular evolution inference based on the partial sequence of lef-8, lef-9 and polh genes shows that LdMNPV-BNP and isolates of Lymantria monacha nucleopolyhedrovirus (LymoNPV) may share a very recent common ancestor or be isolates of the same virus species. LdMNPV-BNP, like other baculoviruses, could be beneficial as an active component of biopesticides that can be used during forest integrated pest management.

Feeding Experience Affects the Behavioral Response of Polyphagous Gypsy Moth Caterpillars to Herbivore-induced Poplar Volatiles.

Plant volatiles influence host selection of herbivorous insects. Since volatiles often vary in space and time, herbivores (especially polyphagous ones) may be able to use these compounds as cues to track variation in host plant quality based on their innate abilities and previous experience. We investigated the behavioral response of naïve (fed on artificial diet) and experienced (fed on poplar) gypsy moth (Lymantria dispar) caterpillars, a polyphagous species, towards constitutive and herbivore-induced black poplar (Populus nigra) volatiles at different stages of herbivore attack. In Y-tube olfactometer assays, both naïve and experienced caterpillars were attracted to constitutive volatiles and volatiles released after short-term herbivory (up to 6 hr). Naïve caterpillars also were attracted to volatiles released after longer-term herbivory (24–30 hr), but experienced caterpillars preferred the odor of undamaged foliage. A multivariate statistical analysis comparing the volatile emission of undamaged plants vs. plants after short and longer-term herbivory, suggested various compounds as being responsible for distinguishing between the odors of these plants. Ten compounds were selected for individual testing of caterpillar behavioral responses in a four-arm olfactometer. Naïve caterpillars spent more time in arms containing (Z)-3-hexenol and (Z)-3-hexenyl acetate than in solvent permeated arms, while avoiding benzyl cyanide and salicyl aldehyde. Experienced caterpillars avoided benzyl cyanide and preferred (Z)-3-hexenyl acetate and the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) over solvent. Only responses to DMNT were significantly different when comparing experienced and naïve caterpillars. The results show that gypsy moth caterpillars display an innate behavioral response towards constitutive and herbivore-induced plant volatiles, but also that larval behavior is plastic and can be modulated by previous feeding experience.Electronic supplementary materialThe online version of this article (doi:10.1007/s10886-016-0698-7) contains supplementary material, which is available to authorized users.

Glutathione S-transferase in the midgut tissue of gypsy moth (Lymantria dispar) caterpillars exposed to dietary cadmium.

Activity of glutathione S-transferase (GST) in midgut of gypsy moth caterpillars exposed to 10 and 30μg Cd/g dry food was examined. Based on the enzyme reaction through conjugation with glutathione, overall activity remained unaltered after acute and chronic treatment. No-observed-effect-concentration (10μg Cd/g dry food) significantly increased activity only after 3-day recovery following cadmium administration. Almost all comparisons of the indices of phenotypic plasticity revealed statistically significant differences. Despite the facts that GST has important role in xenobiotic biotransformation, our results indicate that this enzyme in insect midgut does not represent the key factor in cadmium detoxification.

Tannic acid modulates the activity of bombyxin- and PTTH-producing neurons in Lymantria dispar L. (Insecta: Lepidoptera) caterpillars

Lymantria dispar caterpillars were exposed for 3 days to tannic acid, added to the artificial diet in the following concentrations: 1, 2.5 and 5%. Changes in the morphometric parameters of medial A2 bombyxin immunopositive and dorsolateral L2` prothoracicotropic (PTTH) immunopositive neurosecretory neurons (nsn; sizes of six nsn, the size of their nuclei, and quantification of cytoplasm density) were analysed upon caterpillars’ exposure to this allelochemical. Protein band density, obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) electrophoresis, from regions of bombyxin and PTTH molecular mass (Mr) were estimated. A2 nsn decreased in size, while the size of their nuclei did not change significantly. Relative cytoplasm density and relative band density from the bombyxin Mr region (Mr 4–5 kD) were decreased upon exposure to 1% and 2.5% tannic acid, while the addition of 5% tannic acid to artificial diet increased all of these parameters. Results suggested that lower concentrations of tannic acid stimulate the release of neurosecretory material from A2 nsn, most probably bombyxin-like protein. High tannic acid concentration (5%) probably allocates resources towards induction of other stress protective components instead of bombyxin-mediated metabolic stress response mechanisms. In L2` nsn, the size of nsn, nuclei and the relative cytoplasm density were increased, and relative band density from the region of big PTTH form (Mr 11–15 kD) was higher in all treatments in comparison to control. Results indicate that PTTH is synthesised and stored in the cytoplasm of L2` nsn.