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.

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.

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.

Cardioinhibitory properties of potato glycoalkaloids in beetles.

The semi-isolated heart bioassay was used to evaluate the effect of glycoalkaloids extracted from potato leaves on the heart contractile activity of three beetle species Zophobas atratus, Tenebrio molitor and Leptinotarsa decemlineata. The dose-response curves indicated species specific action of tested substances. Application of glycoalkaloids on the continuously perfused Z. atratus heart inhibited progressively frequency contractions; higher concentrations exerted short and reversible cardiac arrests. In the rest two beetle species tested glycoalkaloids caused no cardiotropic effect. In vivo bioassay with 1 day old Z. atratus pupae showed that the extract induces a negative inotropic effect on the heart.

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.

Non-omnia moriantur-toxicity of mancozeb on dead wood microarthropod fauna.

The effect of Dithane M-45 (dithiocarbamate fungicide; active substance: mancozeb) was studied on microarthropod fauna inhabiting dead wood. Although the exposure was almost never 100% lethal for the majority of observed taxa, almost all (Mesostigmata, Oribatida, some Uropodina, Actinedida, Collembola and Diplopoda) showed very high correlation between concentration of the fungicide and mortality (r > 0.86). Only Stigmaeidae showed low correlation (r = 0.293). For the majority of taxa LC(50 )values were close to the concentrations used during agrochemical activities in woods. Only Trachytes aegrota showed full susceptibility to the fungicide within the range of recommended field concentrations used in forestry (characterised by the low LC(95 )value). Tolerance of mesostigmatid and oribatid mites was found to differ between juveniles and adults, but not consistently. Related Uropodina species varied in susceptibility to the fungicide.

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.

Molecular and phenotypic characterisation of Bacillus thuringiensis isolated during epizootics in Cydia pomonella L.

Twelve Bacillus thuringiensis strains were isolated from intestinal tracts of Cydia pomonella larvae during epizootics in different laboratory insect culture lines. Phenotypic and genetic similarity of these isolates, together with two cultured from Leucoma salicis larvae and 14 reference B. thuringiensis strains were determined. The epizootic bacteria did not form a single group based on numerical analysis of biochemical properties. Simple RAPD method with only one primer does not allow estimating the genetic similarity of B. thuringiensis strains. We propose a novel strategy based on combining several DNA patterns obtained by RAPD technique with different primers for B. thuringiensis typing. Majority of infections in the C. pomonella culture lines were caused by bacteria with different genotypes. However, two isolates cultured from infected insects at different time (one strain was isolated in 1990 and the other in 1992) had identical DNA fingerprint that suggested a possibility of these bacteria to survive in the laboratory and to cause infections in different years. The results of SDS-PAGE of whole-cell proteins revealed a possibility to apply protein profile analysis in epidemiological investigations of infections caused by B. thuringiensis. Strains with identical DNA patterns had very similar whole-cell protein profiles.