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Gypsy moth, Lymantria dispar L. (Lepidoptera: Erebidae) feeds on a large number of tree species, while ash, Fraxinus spp. (Lamiales: Oleaceae) species are considered resistant and are only sporadically eaten. To assess the conditions under which late instar gypsy moth larvae (GML) can temporarily use non-host common ash (CA) (F. excelsior L.), and to evaluate their ability to recover from ingestion of this toxic food, we determined the relative growth rate, the relative consumption rate and the amount of produced feces in different laboratory feeding trials. Our report is the first to show that under specific circumstances, the resources acquired after short-term consumption of CA leaves can be utilized for larval growth. We varied the intensity of density and starvation stress prior to feeding on CA leaves. We observed that after moderate stress a group of GML was temporarily capable of coping with CA leaves. Although observed growth and consumption were much lower on CA than on the optimal host oak, Quercus cerris L. (Fagales: Fagaceae), CA-oak-switched larvae showed the ability to recover from short-term use of a toxic non-host foliage. This suggests that feeding on CA might enable GML to survive under conditions of food shortage. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 43007: Studying climate change and its influence on the environment: impacts, adaptation and mitigation]

Cadmium pollution is becoming an increasing problem, especially in parts of the world that have developed industries. To consider the potentially harmful effects of cadmium, we need to examine changes at all different levels of biological organization. The main goal of this study was to detect a possible change in the percentage of midgut mass relative to larval mass (PMM) and determine the plasticity of this trait and the correlations between midgut enzymes and PMM under stress conditions. Fourth-instar larvae were exposed to acute and chronic effects of two cadmium concentrations, 10 and 30 μg Cd/g dry food, as well as a threeday recovery from chronic treatments. PMM is also an indirect indicator of food consumption and was found to be significantly reduced compared to control in both acute effects and chronic treatment at 30 μg and its three-day recovery. The PMM reduction during acute treatments is a consequence of cadmium action, while in chronic treatment, the genetic factor (egg mass) plays a crucial role in the change of PMM. According to the index of plasticity, distinct phenotypes were not produced. Significant correlations were shown between PMM and trypsin (Tryp) and leucine aminopeptidases (LAP) at acute and chronic treatment with higher cadmium concentrations, while significant correlations between proteases and PMM were detected at lower metal concentrations (Acute10 and Chronic10 and 30 μg Cd/g dry food). In contrast to chronic treatment, egg masses respond more uniformly by reducing PMM during the short-term effect of cadmium. Finally, we can conclude that, as an addition to biochemical and molecular research, PMM can be used for studying the cadmium effects to gain a better insight into the state of the organism under stress conditions. [ABSTRACT FROM AUTHOR]

The bean weevil (Acanthoscelides obtectus, Say) is a serious pest of stored bean seeds. Bean weevil control relies heavily on the use of synthetic insecticides. In the search for a sustainable alternative, the residual contact toxicity and anti-oviposition activity of thyme (Thymus vulgaris L.), rosemary (Rosmarinus officinalis L.) and basil (Ocimum basilicum L.) essential oils as well as their dominant components (thymol, α-pinene, 1,8-cineole and linalool) were tested against A. obtectus adults. Out of the seven tested botanicals, T. vulgaris oil, thymol and linalool exhibited the highest toxic potential (>90% mortality). Females were less susceptible than males. The insecticidal activity of these botanicals was much greater when they were applied on glass compared to direct application to the bean. All tested botanicals reduced oviposition by bean weevil females. T. vulgaris oil, thymol and α-pinene also deterred bean weevil oviposition, as revealed by a two-choice test. Our research shows that T. vulgaris oil and thymol are promising and sustainable alternatives to synthetic pesticides for protecting stored beans against the bean weevil. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. III43010]

Gypsy moth caterpillars were exposed to an increased rearing temperature of 35ºC and diet, supplemented with Cd, a heavy metal pollutant, and tannic acid, a plant secondary metabolite. After 3 days’ exposure to stressors, changes in the number, morphometric parameters of L1 neurosecretory neurons (nsn) (sizes of the nsn and their nuclei), and the quantity of neurosecretory material in the cytoplasm of the neurons were estimated. Acute exposure to the high temperature of 35˚C induced increases in the number of L1 nsn, their size and the size of their nuclei with prolonged exposure time. After acute exposure to different Cd concentrations, the number of L1 nsn was reduced, their size increased and the size of their nuclei decreased. Together with the enhanced relative density of the cytoplasm, our results point to an intensive synthesis and retention of neurosecretory material in the neurons. The relative density of the neurosecretory material in the cytoplasm increased at the thermal treatment, suggesting intensive synthesis and secretory activity in L1 nsn. Caterpillars reared on an artificial substrate with the addition of high concentrations of tannic acid (TA) showed a decreased number of nsn, increased cell size and decreased size of their nuclei. The reduction in the relative density of the cytoplasm led us to conclude that this treatment induced a high synthetic activity of L1 nsn. [Projekat Ministarstva nauke Republike Srbije, br. 173027]

Gypsy moths and powdery mildew play a significant role in oak decline processes. However, information is lacking on the effects on the gypsy moth of impaired tree vitality caused by defoliation or parasite infection. We assessed how pedunculate oak leaves collected from vigorous, declining, and infected trees influenced gypsy moth preference and performance (growth and nutritional indices). We found a negative effect of powdery mildew-infected leaves on gypsy moth performance, while declining trees had positive effects on gypsy moth performance and preference. All examined parameters of larvae fed declining oak leaves were higher than those of larvae fed vigorous oak leaves. Increased growth on declining oak leaves was caused by both higher consumption and more efficient food utilization. The results of this research could help us to better understand multitrophic interactions in complex communities such as oak forests. [Projekat Ministarstva nauke Republike Srbije, br. 43007: Studying climate change and its influence on the environment: impacts, adaptation and mitigation]

Searching for novel molecular biomarkers, we investigated cadmium effects on the ratio of specific activities of lysosomal and total acid phosphatases (ACPLys/ACPTot) in 4th instar gypsy moth larvae. After acute and chronic exposure to 10 and 30 μg Cd/g dry food, as well as after recovery from both concentrations, the trait values, plasticity, variability and genetic correlations were evaluated. The ACPLys/ACPTot ratio decreased during acute and chronic effects of both concentrations. Inhibition during long-term cadmium exposure was irreversible. Indices of phenotypic plasticity for ACPLys/ACPTot ratio were positive for all cadmium treatments. The variability of plasticity was higher after recovery from 10 μg Cd/g dry food, compared to recovery from 30 μg Cd/g dry food. A significant correlation coefficient was calculated between shortterm cadmium treatments. Significant changes in the ACPLys activity fraction during all treatments indicate the examined trait (ACPLys/ACPTot) could be used as a pollution exposure biomarker.

The effects of allelochemical stress on genetic variations in the specific activities of gypsy moth digestive enzymes (trypsin and leucine aminopeptidase) and relative midgut mass (indirect measure of food consumption), as well as variability in their plasticity, were investigated in fifth instar gypsy moths originating from two populations with different trophic adaptations (oak and locust-tree forests). Thirty-two full-sib families from the Quercus population and twenty-six full-sib families from the Robinia population were reared on an artificial diet with or without supplementation with tannic acid. Between population differences were observed as higher average specific activity of trypsin and relative midgut mass in larvae from the Robinia population. Significant broad-sense heritabilities were observed for the specific activity of trypsin in the control state, and for specific activity of leucine aminopeptidase in a stressful environment. Significantly lower heritability for relative midgut mass was recorded in larvae from the Robinia population reared under stressful conditions. Significant variability of trypsin plasticity in larvae from both populations and significant variability of leucine aminopeptidase plasticity in larvae from the Robinia population point to the potential for the evolution of enzyme adaptive plastic responses to the presence of stressor. Non-significant across-environment genetic correlations do not represent a constraint for the evolution of enzyme plasticity. [Projekat Ministarstva nauke Republike Srbije, br. 173027]

The influence of allelochemical stress and population origin on the patterns of phenotypic and genetic correlations among life history traits and digestive enzyme activities were investigated in larvae of the gypsy moth (Lymantria dispar L.; Lepidoptera: Lymantriidae). Thirty-two full-sib families from oak (suitable host plant, Quercus population), and twenty-six full-sib families from locust-tree (unsuitable host plant, Robinia population) forests were reared on an artificial diet, with or without a 5% tannic acid supplement. Comparison of correlation matrices revealed significant similarity between the two populations in the structure of phenotypic and genetic correlations of life history traits and of digestive enzyme activities. The patterns of correlations of the examined traits, within each of the two locally adapted populations and in the presence of allelochemical stress, remained stabile despite the different selection pressures that mold these traits. [Projekat Ministarstva nauke Republike Srbije, br. 173027]

Lymantria dispar caterpillars were reared under two different rearing densities for the first three days of the 4th larval instar: 5 larvae that were kept in a Petri dish (V = 80 ml) belonged to the intense stress (D1 group); 5 larvae that were kept in a plastic cup (V = 300ml) belonged to the group exposed to less intense stress (D2 group). In the control group, single larvae were reared in a Petri dish. Morphometric changes in L1, L2 and L2’ dorsolateral neurosecretory neurons (nsn) were analyzed. After keeping 5 larvae in a Petri dish, the size of L2 neurosecretory neurons (nsn) significantly increased. Rearing 5 larvae in a plastic cup significantly increased the size of L1 nsn nuclei and the number of L2’nsn. A decrease in relative band densities in the region of molecular masses (11-15 kD) that correspond to prothoracicotropic hormones in the gypsy moth was observed in the electrophoretic profiles that were obtained after both treatments in comparison to the control group. [Acknowledgments. This study was supported by the Serbian Ministry of Education and Science (Grant No. 173027).]

To assess the plastic and genetic components of variation in responses of gypsy moth (Lymantria dispar) 4th instar larvae to temperature and food quality, we applied a split-family four-environment experimental design where full-sibs were reared on two constant temperatures (23°C and 28°C) and two concentrations of dietary nitrogen (1.5 and 3.7% dry weight). A temperature of 28°C and low dietary nitrogen decreased larval weight and prolonged larval developmental time, while viability was not affected. Only a marginally significant interaction between the two environmental factors was found for larval weight. The broad-sense heritability for larval developmental time did not change across environments, and across-environment genetic correlations were close to one. Heritability for larval weight depended on environmental and across-environmental genetic correlations that were not significant. There was no evidence of a trade-off between developmental time and larval weight. The implications of the obtained results for the evolution of phenotypic plasticity in complex environments are discussed. [Acknowledgments. This work was supported by Ministry of Education and Science of Serbia, grant No. 173027.]

Gypsy moth 4th instar caterpillars were fed for 3 days with an artificial diet supplemented with increasing cadmium (Cd) concentrations (0, 10, 30, 100 and 250 μg⁄g of dry food weight). Changes in the morphometric characteristics of A1’ dorso-medial and L2 dorso-lateral neurosecretory neurons (nsn) were analyzed. In the A1’ nsn, Cd supplements led to an enhanced nuclear size, except in the group treated with 250 μg Cd⁄g in the form of dry food. The size of L2 type nsn was increased in the groups provided with 30 and 100 μg Cd⁄g, while no differences in the size of nuclei was detected in L2 neurons among the experimental groups.

In this study we investigated the effects of tannic acid on fitness-related traits in gypsy moth larvae from two differently adapted populations. Thirty two full-sib families from oak (suitable host plant, Quercus population), and twenty six full-sib families from locust-tree (unsuitable host plant, Robinia population) forests were reared on artificial diets with or without a 5% tannic acid supplement. Tannic acid shortened the duration of larval development till the 4th instar in the Robinia population, and decreased the mass of larvae from both Quercus and Robinia populations. Local adaptation was not recorded for any of the examined traits. In general, regardless of population origin, genetic variation (broad-sense heritability) was higher in larvae fed on the diet containing tannic acid than in larvae fed on the control diet. Variability of phenotypic plasticity was significant for the examined traits, except for larval duration traits in Quercus larvae, pointing to the potential of gypsy moth larvae from both populations for the evolution of adaptive plastic responses to new environmental conditions and the presence of stressors. Genetic correlations between the environments were positive and significantly different from “one” and accordingly, do not represent constraint for the evolution of plasticity.

The morphometric changes (size of neurons and their nuclei) of protocerebral dorsomedial A1’ and dorsolateral L2’ neurosecretory neurons were analyzed in Lymantria dispar larvae after exposure to strong static (SMF, 235 mT) and extremely low frequency magnetic fields (ELF MF, 2 mT). Increase in the size of A1’ neurons and their nuclei were observed after acute exposure to SMF. Decrease in the size of these neurons and their nuclei was observed after exposure to ELF MF. The size of L2’ neurons and their nuclei tend to decrease after exposure to SMF and ELF MF. The quantification of protein bands within the Mr range corresponding to the large form of the prothoracicotropic neurohormone indicates that the amount of protein decreased after exposure to both types of magnetic fields.

Morphometric changes of A1, A1' and A2 protocerebral dorsomedial neurosecretory neurons, total brain protein content and brain protein profiles were analyzed in 4th instar Lymantria dispar larvae under elevated rearing density, i.e. under intense stress when 5 larvae were kept in a petri dish (V = 80 ml), less intense stress when 5 larvae were kept in a plastic cup (V = 300 ml). In the control samples the larvae were reared in isolated conditions. Protein pattern changes in the brain were observed. Proteins with the following molecular masses: 30, 14, 10 and 3.4-2.5 kD were detected in the experimental groups. The size and cytological characteristics of protocerebral dorsomedial neurosecretory neurons were changed under elevated rearing density.

Periodic fluctuations in density impose different selection pressures on populations of outbreaking Lepidoptera due to changes in their nutritional environment. The maternal effects hypothesis of insect outbreak predicts the transmission of this nutritional "information" to subsequent generations and alterations in offspring life-history traits. To test for these time-delayed effects of the parental generation, we compared life-history traits and their variation and covariation among laboratory-reared gypsy moths hatched from egg masses collected from low- and medium-density populations. Decreased individual performance was recorded in offspring from the medium-density population, indicating reduced egg provisioning under crowding conditions. Genetic variance and covariance were also shown to be sensitive to density of the parental generation. In gypsy moths from the medium-density population, quantitative genetic analysis revealed significantly higher broad-sense heritabilities for development duration traits and demonstrated a trade-off between development duration and body size.

Insect growth and development depend on nutritive value and secondary metabolite content of their host plants. This influence may be exerted through changing the activity of digestive enzymes which further affects efficiency of conversion of ingested food. This paper represents the results of investigation of feeding effects on the leaves of three oak species (Quercus cerris, Q. robur and Q. petraea) on growth, development and digestive enzyme activities of the 4th instar gypsy moth larvae (Lymantria dispar L.). Compared to larvae fed on Q. cerris and Q. robur leaves, larvae fed on Q. petrea exhibited the lowest body mass, midgut protein content and specific protease activity. Specific amylase activity was significantly reduced in the midguts of larvae fed on Q. robur while host plant did not affect larval duration upon molting into the 4th instar. Concomitant chemical analyses of leaves given to the 2nd and 4th instar larvae was carried out in order to explain the obtained results.

The effects of constant temperature and an extremely low-frequency magnetic field (ELFMF, 50 Hz and average induction of 20 mT) on the activity of medial protocerebral neurosecretory neurons (A1 and A2) and corpora allata were investigated in Cerambyx cerdo L. larvae after 30 days of exposure. Both constant temperature of 23°C and the ELFMF led to decrease in activity of A1 and A2 neurosecretory neurons and increase in activity of corpora allata compared to the control group (larvae from natural conditions). The changes are more pronounced in A2 than A1 neurons.

The effects of different temperatures (23°C and 8°C) on activity of corpora allata (CA) and dorsolateral (L1, L2) protocerebral neurosecretory neurons were investigated in Morimus funereus Mulsant (1863) larvae collected from a natural population during March. Activity of CA was revealed by monitoring of CA volume and cell number. Increase of CA volume after two day exposure to both temperatures was shown to be the result of increase in cell number. Activity of CA was higher at 23°C than 8°C. Activity of L1 and L2 neurosecretory neurons was inhibited at both temperatures. Neurosecretory neurons were more sensitive to temperature of 23°C than 8°C. It can be supposed that dorsolateral neurosecretory neurons synthesize neurohormones that affect CA activity, depending on environmental temperature.

The response of starved Morimus funereus larvae refed with an artificial diet (Galford, 1967) was examined in the present paper. Larvae were offered diets varying in protein quality (soya protein, casein, and gelatin). Refeeding with the Galford diet (G I) and two modifications of it in which soya protein was supplemented with casein (G II) or gelatin (G III) led to an increase of body mass. Different protein quality in the nutritive substrate influenced both the quantity and quality of midgut and brain proteins.