Kazimierz Ziemnicki

Analysis of cry Gene Profiles in Bacillus Thuringiensis Strains Isolated During Epizootics in Cydia pomonella L.

The crystal morphology and the profiles of genes encoding protein toxins (Cry and Cyt) were analyzed in 12 Bacillus thuringiensis strains isolated during epizootics in laboratory culture lines of Cydia pomonella, 2 isolates cultured from Leucoma salicis larvae, and 9 reference strains. Epizootic isolates produced crystals of the same bipyramidal shape; however, they revealed a variety of number and type of cry genes. Genes cry1I, cry2Ab, and cry9B were the most frequently observed in epizootic strains. Gene cry1I was noted in of 50% epizootic isolates. Eighty-three percent of them harbored gene cry2Ab. Gene cry9B was found for 42% of strains isolated during epizootics. Three isolates showed the largest number of cry genes and their variety; hence, they were chosen for the toxicity assay of their crystals and spores on C. pomonella larvae. One of them had approximately sixfold higher insecticidal activity than the reference strain B. thuringiensis subsp. kurstaki BTK STANDARD.

Effect of boric acid on antioxidant enzyme activity, lipid peroxidation, and ultrastructure of midgut and fat body of Galleria mellonella

Boric acid is widely used as an insecticide, acaricide, herbicide, and fungicide and also during various industrial processings. Hence, numerous populations are subjects to this toxic compound. Its action on animals is still not fully known and understood. We examined the effect of boric acid on larvae of greater wax moth (Galleria mellonella). The chemical appeared to be toxic for larvae, usually in a concentration-dependent manner. Exposed groups revealed increased lipid peroxidation and altered activity of catalase, superoxide dismutase, glutathione S-transferase, and glutathione peroxidase. We also observed changes of ultrastructure, which were in tune with biochemical assays. We suggest that boric acid has a broad mode of action, which may affect exposed larvae, and even if sublethal, they may lead to disturbances within exposed populations.

Inheritance of chorionic malformations and insecticide resistance by Spodoptera exigua

Abstract:  Effect of exposure to the organophosphorus insecticide – fenitrothion [O,O‐dimethyl‐O‐(4‐nitro‐3‐methyl)phenyl] – on four generations of Spodoptera exigua Hübner (Lep., Noctuidae) larvae, with regard to the mortality and structure of the chorion of eggs laid was tested. Generations varied in the total mortality rate, although all of them showed some survival rate. The third and the fourth generations showed the lowest mortality among tested ones. Exposure to fenitrothion caused malformations in eggs. The chorion revealed cracks and diminutions of the outer layer. The quality and quantity of malformations increased proportional to the duration of exposure, e.g. the second exposed generation reveald more prominent changes than the first one. Malformations were observed in the next generations, which were not exposed to pesticide. The follicular cells, which are responsible for the structure and sculpture of the eggshell, must have inherited the malforming mechanism. Most probably, malformation takes place during the late development of eggs in the ovarioles.

The influence of dietary α-solanine on the waxmoth Galleria mellonella L.

Plant allelochemicals are nonnutritional chemicals that interfere with the biology of herbivores. We posed the hypothesis that ingestion of a glycoalkaloid allelochemical, α-solanine, impairs biological parameters of greater wax moths Galleria mellonella. To test this idea, we reared wax moths on artificial diets with 0.015, 0.15, or 1.5 mg/100 g diet of α-solanine. Addition of α-solanine to the diet affected survival of seventh-instar larvae, pupae, and adults; and female fecundity and fertility. The diet containing the highest α-solanine concentration led to decreased survivorship, fecundity, and fertility. The diets supplemented with α-solanine led to increased malondialdehyde and protein carbonyl contents in midgut and fat body and the effect was dose-dependent. Dietary α-solanine led to increased midgut glutathione S-transferase activity and to decreased fat body glutathione S-transferase activitiy. We infer from these findings that α-solanine influences life history parameters and antioxidative enzyme activities in the midgut and fat body of G. mellonella.

Effect of Various Xenobiotics on Hatching Success of Spodoptera exigua Eggs as Compared to a Natural Plant Extract

The effects of fenitrothion, carbaryl, and mancozeb, present in polluted water and plant extracted glycoalkaloids, were examined on hatching success of Spodoptera exigua eggs. All chemicals produced a significant decrease in hatching success, which was correlated with chemical concentration. One of the most interesting aspects of this study relates to the biological activity of glycoalkaloids.

POTATO LEAF EXTRACT AND ITS COMPONENT, α‐SOLANINE, EXERT SIMILAR IMPACTS ON DEVELOPMENT AND OXIDATIVE STRESS IN Galleria mellonella L.

Plants synthesize a broad range of secondary metabolites that act as natural defenses against plant pathogens and herbivores. Among these, potato plants produce glycoalkaloids (GAs). In this study, we analyzed the effects of the dried extract of fresh potato leaves (EPL) on the biological parameters of the lepidopteran, Galleria mellonella (L.) and compared its activity to one of the main EPL components, the GA α-solanine. Wax moth larvae were reared from first instar on a diet supplemented with three concentrations of EPL or α-solanine. Both EPL and α-solanine affected survivorship, fecundity, and fertility of G. mellonella to approximately the same extent. We evaluated the effect of EPL and α-solanine on oxidative stress in midgut and fat body by measuring malondialdehyde (MDA) and protein carbonyl (PCO) contents, both biomarkers of oxidative damage. We evaluated glutathione S-transferase (GST) activity, a detoxifying enzyme acting in prevention of oxidative damage. EPL and α-solanine altered MDA and PCO concentrations and GST activity in fat body and midgut. We infer that the influence of EPL on G. mellonella is not enhanced by synergistic effects of the totality of potato leaf components compared to α-solanine alone.

Interaction between crystalline proteins of two Bacillus thuringiensis strains against Spodoptera exigua

The aim of the present article was to evaluate potential synergism between crystalline proteins produced by two Bacillus thuringiensis Berliner strains, MPU B6 and MPU B9, against beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Protein inclusions of bacterial strains were isolated from a spore‐crystal mixture. We estimated the 50% lethal concentration (LC50) of crystals for S. exigua larvae. Insecticidal activity of MPU B6 and MPU B9 individual crystal preparations against caterpillars were determined and compared with the commercial pesticide Foray. Protein crystals of MPU B9 had the highest toxicity against S. exigua. The proteins were approximately 25× more toxic than Foray. Insecticidal activity of protein crystals of MPU B6 isolate was approximately 2.5× higher than that of Foray. A mixture of crystals suspensions of both isolates MPU B6/MPU B9 had an additive effect on S. exigua caterpillars. The high insecticidal potency of B. thuringiensis MPU B9 crystals against S. exigua predisposes the strain for additional studies on production of a new effective preparation against pest insects.

Diflubenzuron inhibits reproduction of different strains of Drosophila melanogaster.

Abstract  The effect of (1‐(4‐chlorophenyl)‐3‐(2,6‐difluorobenzoyl)urea) insecticide – diflubenzuron – on wild type and white type fruit flies Drosophila melanogaster (Meigen, 1830) was studied. Adult insects were placed in vials with different concentrations of the insecticide in the nutrient (from 0.048 to 48 mg of diflubenzuron per 1 cm3 of the nutrient). In each case, the insects showed some mortality, which was concentration‐dependant. When comparing both strains, we could observe different acute toxicities, with wild type being more resistant to diflubenzuron. However, subacute toxicity was similar within both strains. In both strains the prolonged exposure led to the extinction of the majority of the populations (>70%). Although imagoes could freely copulate, we did not observe copulating fruit flies, and we did not find either eggs or larvae in any of the exposed groups. These results indicate that genetic mutation which leads to different eye colour can also affect resistance and survival of insects in pesticide‐exposed areas.

Effect of fenitrothion on Spodoptera exigua larval development and ultrastructure of follicle cells.

The effect of four sublethal concentrations (from 0.01165 to 0.3025 µg per insect, responding to LC10/10, LC10, LC30 and LC50) of the organophosphorus insecticide, fenitrothion, on Spodoptera exigua (Lepidoptera, Noctuidae) larval development and ultrastructure of follicular cells of adult female moths was tested. The most prominent malformations were: breaks in the cuticle, increased mortality during larval/pupal molting, alterations of nuclear envelope, changed heterochromatin pattern and abnormalities in mitochondrial ultrastructure. Some of the alterations are dose-dependent, but the other ones, like organization of rough endoplasmic reticulum (RER), show non-linear response: cells exposed to lower concentrations (LC10/10 and LC10) of the insecticide possessed clusters of accumulated RER, which were absent within the cells exposed to higher ones (LC30 and LC50). Formation of the eggshell was also markedly postponed within the groups exposed to lower doses. These findings prove that even minute amounts of xenobiotics may cause significant changes within exposed populations.