Sex specificity in innate immunity of insect larvae

The innate immunity of insects has been widely studied. Although the effect of sex on insect immunity has been extensively discussed, differences in immunity between the sexes of larvae insects remain largely unstudied. Studying larval sex differences in immunity may provide valuable information about the mechanisms underlying the insect immune system, which, in turn, can be valuable for the development and improvement of pest management. Here we compared the antibacterial activity in both the midgut tissue and cell-free hemolymph of Lymantria dispar L. (Lepidoptera: Erebidae) females and males at the larval stage without and after a challenge by entomopathogenic bacterium Bacillus thuringiensis Berliner. We also evaluated the sex-specific mortality of L. dispar induced by B. thuringiensis infection. We find that antibacterial activity in the midgut is activated by infection, but only in females. Thus, sex differences in immunity can have important effects even before sexual differentiation at adulthood

Molecular sexing of Lepidoptera

We developed a universal method of Lepidoptera molecular sexing. The method is based on comparing the number of copies of the same gene in different sexes. Males of the majority of lepidopteran species have two Z chromosomes, whereas females have only one Z chromosome. Correspondingly, the number of copies of each gene located on this chromosome differs by two times between males and females. For quantitative estimation, we used qPCR. Via multiple alignment of the kettin (a Z chromosome gene) nucleotide sequences, we detected the most conserved fragment and designed primers with broad interspecies specificity for Lepidoptera. Using these primers, we successfully determined the sex of three lepidopteran species belonging to different superfamilies. The developed method is a simple, cost-effective and high-throughput technique for routine sexing. The sex of lepidopteran individuals can be examined at any developmental stage

A Comparison of the Vertical Transmission of High- and Low-Virulence Nucleopolyhedrovirus Strains in Lymantria Dispar L.

Baculoviruses can persist in insect host organisms after infection and may be vertically transmitted to the next generation, in which they may be reactivated. The goal of the present study was to compare the efficiency of the vertical transmission of high- and low-virulence strains and the subsequent reactivation of Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) in the offspring of Lymantria dispar L. adults who survived after viral infection. As a result of parental infection, the fecundity of survived females, pupae weight, and fertility were significantly different compared to the untreated insects. However, differences in these parameters between high- and low-virulence strains were not observed. The prevalence of virus strains in the offspring measured by quantitative polymerase chain reaction also did not differ. When the larvae reached the fourth instar, they were starved to activate the vertically transmitted virus. The frequency of virus activation in the experiment was not dependent on the virulence of the virus strains. These results are helpful for understanding the strategy of virus survival in nature and for the selection of the most effective strains with transgenerational effects in the years following pest treatment

Asynchrony between host plant and insects-defoliator within a tritrophic system: The role of herbivore innate immunity

The effects of asynchrony in the phenology of spring-feeding insect-defoliators and their host plants on insects’ fitness, as well as the importance of this effect for the population dynamics of outbreaking species of insects, is a widespread and well-documented phenomenon. However, the spreading of this phenomenon through the food chain, and especially those mechanisms operating this spreading, are still unclear. In this paper, we study the effect of seasonally declined leafquality (estimated in terms of phenolics and nitrogen content) on herbivore fitness, immune parameters and resistance against pathogen by using the silver birch Betula pendula—gypsy moth Lymantria dispar—nucleopolyhedrovirus as the tritrophic system. We show that a phenological mismatch induced by the delay in the emergence of gypsy moth larvae and following feeding on mature leaves has negative effects on the female pupal weight, on the rate of larval development and on the activity of phenoloxidase in the plasma of haemolymph. In addition, the larval susceptibility to exogenous nucleopolyhydrovirus infection as well as covert virus activation were both enhanced due to the phenological mismatch. The observed effects of phenological mismatch on insect-baculovirus interaction may partially explain the strong and fast fluctuations in the population dynamics of the gypsy moth that is often observed in the studied part of the defoliator area. This study also reveals some indirect mechanisms of effect related to host plant quality, which operate through the insect innate immune status and affect resistance to both exogenous and endogenous virus

The effect of population density of Lymantria dispar (Lepidoptera: Erebidae) on its fitness, physiology and activation of the covert nucleopolyhedrovirus

After high population densities of insect defoliators there is often a dramatic decrease in their abundance due to various limiting factors. Here, we compared gypsy moth larvae (Lymantria dispar L.) reared singly and in crowded conditions. We compared a number of physiological parameters of these insects and the effect of L. dispar population density on the activation of covert baculovirus infections in the larvae. It was found that the population density of gypsy moth larvae did not affect the mortality due to the activation of the covert virus infection or the total mortality. On the other hand, solitary-reared larvae were heavier, took longer to develop, and showed a four-fold higher concentration of dopamine in their haemolymph than larvae reared in groups. Thus, we demonstrated that an increase in the population density of larvae per se facilitates some changes in fi tness and innate immunity traits but is not related to the activation of covert baculovirus infection. We suggest that an increase in population density does not increase the risk of epizootics triggered by the activation of covert baculovirus infection and that researchers should pay more attention to studying density-associated factors, such as starvation

The effect of mixtures of Bacillus thuringiensis-based insecticide and multiple nucleopolyhedrovirus of Lymantria dispar L. in combination with an optical brightener on L. dispar larvae

This study evaluated the efficacy of the commercially available insecticide Lepidocide based on Bacillus thuringiensis var. kurstaki and Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) and their combination with an optical brightener to control L. dispar L. Efficacy against both second and fourth instar L. dispar larvae was evaluated, and the type of interaction between the tested components was determined using second instar L. dispar larvae. Most combinations of Lepidocide and LdMNPV containing a 5 mg ml(-1) optical brightener had synergistic effects, and their mixtures were most effective in reducing the number of second instar larvae. In contrast, mixtures containing Lepidocide and LdMNPV with an optical brightener caused significantly lower mortality of fourth instar L. dispar larvae than mixtures without Lepidocide. This result suggests that an increased concentration of Lepidocide in a mixture containing LdMNPV and an optical brightener leads to an antagonistic effect on insect mortality. The possible reasons for the differences in the observed effects of the components on the second and fourth instar L. dispar larvae may be associated with the resistance of fourth-instar larvae to the antifeedant effect of B. thuringiensis

Leaf Surface Lipophilic Compounds as One of the Factors of Silver Birch Chemical Defense against Larvae of Gypsy Moth

<div><p>Plant chemical defense against herbivores is a complex process which involves a number of secondary compounds. It is known that the concentration of leaf surface lipophilic compounds (SLCs), particularly those of flavonoid aglycones are increased with the defoliation treatment of silver birch <i>Betula pendula</i>. In this study we investigated how the alteration of SLCs concentration in the food affects the fitness and innate immunity of the gypsy moth <i>Lymantria dispar</i>. We found that a low SLCs concentrations in consumed leaves led to a rapid larval development and increased females’ pupae weight (= fecundity) compared to larvae fed with leaves with high SLCs content. Inversely, increasing the compounds concentration in an artificial diet produced the reverse effects: decreases in both larval weight and larval survival. Low SLCs concentrations in tree leaves differently affected larval innate immunity parameters. For both sexes, total hemocytes count in the hemolymph increased, while the activity of plasma phenoloxidase decreased when larvae consume leaves with reduced content of SLCs. Our results clearly demonstrate that the concentration of SLCs in silver birch leaves affects not only gypsy moth fitness but also their innate immune status which might alter the potential resistance of insects against infections and/or parasitoids.</p></div

Effect of <i>Betula pendula</i> leaf surface lipophilic compounds on <i>Lymantria dispar</i> larval weight.

<p>The weight (mean ±SE) of young (a) and middle (b) instar larvae reared on leaves with low and high concentrations of surface lipophilic compounds is presented. The data were compared using a one-way ANOVA. Asterisk means the significant differences between bars (at <i>P</i><0.05).</p

Effect of <i>Betula pendula</i> leaf surface ethanol extract on <i>Lymantria dispar</i> fitness.

<p>Weight (mean ±SE) (a) and survival rate (mean ±SE) (b) of <i>Lymantria dispar</i> larvae reared on artificial diet with different concentrations of leaf surface extract is presented. The data were pair-wise compared by post hoc Fisher LSD procedure. The letters above the bar mean the significant differences (at <i>P</i><0.05) to be compared with the bars abbreviated by the same letters within bar.</p

Factorial ANOVA results of comparison of pupae weight between females and males reared on <i>Betula pendula</i> leaves with low and high concentrations of surface lipophilic compounds.

<p>Factorial ANOVA results of comparison of pupae weight between females and males reared on <i>Betula pendula</i> leaves with low and high concentrations of surface lipophilic compounds.</p