Changes in the Activity and Pattern of Hemolymph Esterases in the Larvae of Greater Wax Moth Galleria mellonellaL. (Lepidoptera, Pyralidae) during Mycosis

The extended pattern of multiple esterase forms has been revealed in the hemolymph of wax moth Galleria mellonellalarvae infected by the fungi Metarhizium anisopliae, Beauveria bassianaor Paecilomyces fumoso-roseus. The total esterase activity of the hemolymph also increases during mycosis. Mechanical damage of the cuticle, treatment with deltamethrin, and chilling of the caterpillars induced similar changes in the hemolymph pattern of esterase activity. Presumably, the changed spectrum and activity of the hemolymph esterases during mycosis is due to the damaged cuticle and epidermis cells.     

Model of dynamics of the insect cellular immune system

A mathematical model for the dynamics of functioning of an insect immune subsystem is considered. It is assumed that the interaction between hemocytes and bacteria corresponds to the Volterra principle of "pair interactions": the rate at which bacteria enter the system decreases in proportion to the rate of interaction of hemocytes with bacteria. In the absence of interaction, the intrinsic dynamics of bacteria is described by the Malthus law. Dynamic regimes of the model for different values of the parameters were analyzed. In particular, it was shown that, depending on the initial values of variables and the values of the parameters, there exist two regimes, the regime of death of the organism and the regime of elimination of bacteria.     

VENOM FROM THE ECTOPARASITIC WASP Habrobracon hebetor ACTIVATES CALCIUM‐DEPENDENT DEGRADATION OF Galleria mellonella LARVAL HEMOCYTES

Ectoparasitoids inject venom into hemolymph during oviposition. We determined the influence of envenomation by the parasitoid, Habrobracon hebetor, on the hemocytes of its larval host, Galleria mellonella. An increase in both intracellular Са²⁺ content and phospholipase C activity of the host hemocytes was recorded during 2 days following envenomation by the parasitoid. The decreased hemocyte viability was detected 1, 2, and 24 h after the envenomation. Injecting of the crude venom (final protein concentration 3 μg/ml) into the G. mellonella larvae led to the reduced hemocyte adhesion. The larval envenomation caused a decrease in transmembrane potential of the hemocytes. These findings document the suppression of hemocytic immune effectors in the parasitized host larvae.     

Generation of reactive oxygen species and activity of antioxidants in hemolymph of the moth larvae <Emphasis Type="Italic">Galleria mellonella</Emphasis> (L.) (Lepidoptera: Piralidae) at development of the process of encapsulation

Activities of enzymatic antioxidants—superoxide dismutase, glutathione-S-transferase, and catalase—as well as generation of reactive oxygen species (ROS) in lymph of the honeycomb moth Galleria mellonella L. were studied at development of the process of encapsulation of nylon implants. It has been established that as soon as 15 min after piercing of cuticle with the implant the capsule is formed on its surface. Active melanization of the capsule has been shown to last for 4 h. A statistically significant increase of the ROS generation in lymph and a decrease of the enzymatic antioxidant activities in the insect hemocytes have been revealed after the implant incorporation. The authors suggest that the key role in maintenance of the oxidation-reduction balance in hemolymph at development of the encapsulation process is played by non-oxidative antioxidants.     

Diversity of microbial communities of the intestinal mucosa and intestinal contents of fish from Lake Chany (Western Siberia)

The diversity of microbial communities associated with the intestinal mucosa and the intestinal contents of fish with different nutritional profiles have been investigated. Differences in the intestinal microbiota of fish with different dietary preferences have been revealed using group-specific primers for the main large taxa of bacteria. Representatives of the phyla Planctomycetes, Vernicomicrobia, and Cyanobacteria have been detected in the intestinal contents of all investigated nonpredatory (crucian carp, roach, and dace) and predatory (pikeperch, pike, and perch) fish. Representatives of the phylum Firmicutes were detected in the intestinal mucosa and intestinal contents of predatory fish species, but not in those of the nonpredatory fish.     

Effect of entomopathogenic fungus <Emphasis Type="Italic">Metarhizium robertsii</Emphasis> on non-target organisms,water bugs (Heteroptera: Corixidae,Naucoridae, Notonectidae)

The influence of the fungus Metarhizium robertsii Bischoff, Rehner and Humber on the mortality of four water bug species, Cymatia coleoptrata (Fabricius), Sigara assimilis (Fieber), Ilyocoris cimicoides cimicoides (Linnaeus), and Notonecta reuteri Hungerford, and bloodsucking mosquito Anopheles messeae Falleroni, was investigated under various concentrations of conidia and different treatment types. We found that the mortality of adults of the water bug species was similar or higher than that of A. messeae, with C. coleoptrata and S. assimilis being more susceptible to M. robertsii than N. reuteri, I. c. cimicoides, and the mosquito A. messeae. Treatment with dry conidia at concentrations of 5 × 10⁴ and 5 × 10⁵ conidia/ml caused higher mortality of the water bug species than did treatment at the same concentrations with conidia in an aqueous suspension. In contrast, higher concentrations (5 × 10⁶ conidia/ml) led to higher mortality after treatment with the aqueous suspension, relative to treatment with dry conidia. Our studies showed that water bugs exhibited the classical development of a mycosis with hemocoel colonization, mummification, and conidia formation on cadavers directly on the surface of the water. Possible changes in invertebrate communities in aquatic ecosystems after treatment with Metarhizium are discussed.     

Insecticidal and immunosuppressive effect of ascomycete Cordyceps militaris on the larvae of the Colorado potato beetle Leptinotarsa decemlineata

The immunosuppressive and insecticidal activity of culture of the entomopathogenic fungus Cordyceps militaris on the larvae of the Colorado potato beetle Leptinotarsa decemlineata has been established for the first time. It was found that the peroral effect of the fungal culture resulted in dose-dependent decrease in survival, delayed in development time and molting, decreases in the total hemocyt counts, increased activity of phenoloxidases in the hemolymph, and reduced activity of the enzyme in the cuticle, as well as increased sensitivity of larvae to the fungus Beauveria bassiana at the level of the synergistic effect.     

Multiplex PCR screening to detect cry9 genes in Bacillus thuringiensis strains.

An extended PCR method was established to rapidly identify and classify Bacillus thuringiensis strains containing cry (crystal protein) genes toxic to lepidopteran, coleopteran, and dipteran pests (Ben-Dov et al., Appl. Environ. Microbiol. 63:4883-4890, 1997). To optimize identification of all reported cry genes, this methodology needs a complete PCR set of primers. In the study reported here, a set of universal (Un9) and specific primers for multiplex rapid screening for all four known genes from the cry9 group was designed. PCR analyses were performed for cry9 genes on 16 standard strains and 215 field isolates of B. thuringiensis. Among the standard strains, only B. thuringiensis subsp. aizawai HD-133, which harbors cry1 and cry2 genes, was positive with Un9 but negative to all four specific primers for cry9 genes. DNA of 22 field-collected isolates was also found to be positive with Un9. These isolates were classified into three cry9 profiles using specific primers; all of them harbor cry1 and cry2. This newly designed set of primers complements the existing PCR methodology for most currently known cry genes.     

Physiological and biochemical aspects of interactions between insect parasitoids and their hosts

In the present review, available literary data on physiological and biochemical interactions between parasitoids and their hosts are analyzed. In order to achieve successful development inside or on their hosts, parasitoids widely use various strategies aimed at suppressing host immunity. Suppression agents used by parasitoids include venom and ovarian fluid components as well as symbiotic microorganisms. The influence of parasitoids on the host organism is complicated, covering many physiological functions and inducing changes of the host metabolism and behavior. The influence of ecto- and endoparasitoids on the host organism is analyzed separately.