Działanie ekstraktu z Solanum nigrum L. i jego głównych glikoalkaloidów, jako czynników zwiększających toksyczność insektycydu fenitrotionu w wybranych tkankach Tenebrio molitor L.

Wydział BiologiiDuring the research, the influence of the black nightshade (Solanum nigrum) unripe fruit extract and its main glycoalkaloids on selected physiological aspects of a yellow mealworm (Tenebrio molitor) was checked for potential usage against this pest. As a next step, the toxic concentrations of fenitrotion (LC50 and LC40) were calculated for the description of the influence of the S. nigrum extract on the toxicity of fenitrothion in two strategies of application. In the first strategy, the extract was mixed with fenitrothion in the ratio 1:1 and applied to the larvae, and in the second strategy, the extract was given 24 h before fenitrothion application. The results showed, that the S. nigrum extract causes sublethal effects such as changes in the activity of visceral muscles, changes in the concentration of stored biomolecules, and disturbances in the midgut and fat body cells in T. molitor. Glycoalkaloids present in the S. nigrum extract cause sublethal toxic effects in yellow mealworm. The differences in the effects caused by the extract and single glycoalkaloids indicate the cooperative action of both glycoalkaloids. The S. nigrum extract applied at the same time with fenitrothion did not increase the lethality of T. molitor larvae comparing to the effects caused by fenitrothion, however, it caused sublethal effects. Appropriate correlation of the S. nigrum extract application before fenitrothion caused significant increase of T. molitor mortality and strengthen the sublethal effects.W niniejszej rozprawie doktorskiej zbadany został wpływ ekstraktu z niedojrzałych owoców psianki czarnej (Solanum nigrum) oraz jego dwóch głównych glikoalkaloidów na wybrane aspekty fizjologii szkodnika magazynów zbożowych, mącznika młynarka (Tenebrio molitor) pod kątem możliwości wykorzystania go w zwalczaniu tego owada. Następnie określono wpływ ekstraktu na toksyczność fenitrotionu (LC50 i LC40) w dwóch strategiach aplikacji. W pierwszej ekstrakt został podany w mieszaninie z fenitrotionem w proporcji 1:1, natomiast w drugiej, ekstrakt został podany na 24 h przed aplikacją fenitrotionu. Otrzymane wyniki wskazały, że ekstrakt z S. nigrum powoduje u mącznika młynarka efekty subletalne, takie jak zmiany w kurczliwości narządów wewnętrznych, zmiany zawartości zmagazynowanych biocząsteczek i zaburzenia w ultrastrukturze komórek jelita środkowego i ciała tłuszczowego. Glikoalkaloidy obecne w ekstrakcie powodują subletalne efekty toksyczne, a różnice w ich działaniu z ekstraktem, wskazują na synergistyczne działanie glikoalkaloidów. Ekstrakt z S. nigrum podawany równocześnie z fenitrotionem nie zwiększa śmiertelności larw, w porównaniu do efektów powodowanych przez sam fenitrotion, jednak widoczne są efekty subletalne. Odpowiednio skorelowane podanie ekstraktu z psianki czarnej przed aplikacją fenitrotionu, spowodowało istotny wzrost śmiertelności T. molitor i nasiliło efekty subletalne

Solanum nigrum Fruit Extract Modulates Immune System Activity of Mealworm Beetle, Tenebrio molitor L

: Here, we report the first evidence concerning the modulation of insect immune system activity after applying Solanum nigrum fruit extract (EXT). We focused on two main issues: (1) is EXT cytotoxic for Tenebrio molitor haemocytes? and (2) how EXT affects the basic immune mechanisms of T. molitor. The results indicate cytotoxic action of 0.01 and 0.1% EXT on beetle haemocytes. Both the injection of EXT and incubating haemocytes with the EXT solution on microscopic slides significantly increased the number of apoptotic cells. However, 24 h after injection of 0.1% EXT cytotoxic effect of the tested extract probably was masked by the increased number of circulating haemocytes. Application of 0.01 and 0.1% EXT led to impairment of the activity of basic immune mechanisms such as phenoloxidase activity and the lysozyme-like antimicrobial activity of T. molitor haemolymph. Moreover, the EXT elicited significant changes in the expression level of selected immune genes. However, some of the immunomodulatory effects of EXT were different in beetles with and without an activated immune system. The obtained results are an essential step toward a complete understanding of the EXT mode of action on the T. molitor physiology and its potential usage in pest control

Solanum nigrum extract and solasonine affected hemolymph metabolites and ultrastructure of the fat body and the midgut in Galleria mellonella

Glycoalkaloids, secondary metabolites abundant in plants belonging to the Solanaceae family, may affect the physiology of insect pests. This paper presents original results dealing with the influence of a crude extract obtained from Solanum nigrum unripe berries and its main constituent solasonine on the physiology of Galleria mellonella (Lepidoptera). Both extract and solasonine applied in the nourishment changed the level of hemolymph metabolites such as sugars, polyols and proline, and ultrastructure of the fat body and the midgut cells. The extract and solasonine, tested in the same concentration found in the extract, decreased the proline level, while mannitol was under the limit of detection in the hemolymph. The extract increased the erythritol level in the hemolymph compared to control, enlarged the intracellular space in fat body cells, decreased cytoplasm and lipid droplets electron density. In the midgut, vacuolized areas of cytoplasm ap-peared with vast reticulum systems. Solasonine, tested with three concentrations, caused the decrease of cytoplasm electron density in both fat body and midgut cells. Obtained results high-lighted the disturbance of the midgut and the fat body due to glycoalkaloids and the potential role of hemolymph ingredients in its detoxification

Solanum nigrum Extract and Solasonine Affected Hemolymph Metabolites and Ultrastructure of the Fat Body and the Midgut in Galleria mellonella

Glycoalkaloids, secondary metabolites abundant in plants belonging to the Solanaceae family, may affect the physiology of insect pests. This paper presents original results dealing with the influence of a crude extract obtained from Solanum nigrum unripe berries and its main constituent, solasonine, on the physiology of Galleria mellonella (Lepidoptera) that can be used as an alternative bioinsecticide. G. mellonella IV instar larvae were treated with S. nigrum extract and solasonine at different concentrations. The effects of extract and solasonine were evaluated analyzing changes in carbohydrate and amino acid composition in hemolymph by RP-HPLC and in the ultrastructure of the fat body cells by TEM. Both extract and solasonine changed the level of hemolymph metabolites and the ultrastructure of the fat body and the midgut cells. In particular, the extract increased the erythritol level in the hemolymph compared to control, enlarged the intracellular space in fat body cells, and decreased cytoplasm and lipid droplets electron density. The solasonine, tested with three concentrations, caused the decrease of cytoplasm electron density in both fat body and midgut cells. Obtained results highlighted the disturbance of the midgut and the fat body due to glycoalkaloids and the potential role of hemolymph ingredients in its detoxification. These findings suggest a possible application of glycoalkaloids as a natural insecticide in the pest control of G. mellonella larvae

Działanie ekstraktu z Solanum nigrum L. i jego głównych glikoalkaloidów, jako czynników zwiększających toksyczność insektycydu fenitrotionu w wybranych tkankach Tenebrio molitor L.

Wydział BiologiiDuring the research, the influence of the black nightshade (Solanum nigrum) unripe fruit extract and its main glycoalkaloids on selected physiological aspects of a yellow mealworm (Tenebrio molitor) was checked for potential usage against this pest. As a next step, the toxic concentrations of fenitrotion (LC50 and LC40) were calculated for the description of the influence of the S. nigrum extract on the toxicity of fenitrothion in two strategies of application. In the first strategy, the extract was mixed with fenitrothion in the ratio 1:1 and applied to the larvae, and in the second strategy, the extract was given 24 h before fenitrothion application. The results showed, that the S. nigrum extract causes sublethal effects such as changes in the activity of visceral muscles, changes in the concentration of stored biomolecules, and disturbances in the midgut and fat body cells in T. molitor. Glycoalkaloids present in the S. nigrum extract cause sublethal toxic effects in yellow mealworm. The differences in the effects caused by the extract and single glycoalkaloids indicate the cooperative action of both glycoalkaloids. The S. nigrum extract applied at the same time with fenitrothion did not increase the lethality of T. molitor larvae comparing to the effects caused by fenitrothion, however, it caused sublethal effects. Appropriate correlation of the S. nigrum extract application before fenitrothion caused significant increase of T. molitor mortality and strengthen the sublethal effects.W niniejszej rozprawie doktorskiej zbadany został wpływ ekstraktu z niedojrzałych owoców psianki czarnej (Solanum nigrum) oraz jego dwóch głównych glikoalkaloidów na wybrane aspekty fizjologii szkodnika magazynów zbożowych, mącznika młynarka (Tenebrio molitor) pod kątem możliwości wykorzystania go w zwalczaniu tego owada. Następnie określono wpływ ekstraktu na toksyczność fenitrotionu (LC50 i LC40) w dwóch strategiach aplikacji. W pierwszej ekstrakt został podany w mieszaninie z fenitrotionem w proporcji 1:1, natomiast w drugiej, ekstrakt został podany na 24 h przed aplikacją fenitrotionu. Otrzymane wyniki wskazały, że ekstrakt z S. nigrum powoduje u mącznika młynarka efekty subletalne, takie jak zmiany w kurczliwości narządów wewnętrznych, zmiany zawartości zmagazynowanych biocząsteczek i zaburzenia w ultrastrukturze komórek jelita środkowego i ciała tłuszczowego. Glikoalkaloidy obecne w ekstrakcie powodują subletalne efekty toksyczne, a różnice w ich działaniu z ekstraktem, wskazują na synergistyczne działanie glikoalkaloidów. Ekstrakt z S. nigrum podawany równocześnie z fenitrotionem nie zwiększa śmiertelności larw, w porównaniu do efektów powodowanych przez sam fenitrotion, jednak widoczne są efekty subletalne. Odpowiednio skorelowane podanie ekstraktu z psianki czarnej przed aplikacją fenitrotionu, spowodowało istotny wzrost śmiertelności T. molitor i nasiliło efekty subletalne

Solanum nigrum Fruit Extract Increases Toxicity of Fenitrothion—A Synthetic Insecticide, in the Mealworm Beetle Tenebrio molitor Larvae

Synthetic insecticides are widely used for crop protection both in the fields and in the food stored facilities. Due to their toxicity, and assumptions of Integrated Pest Management, we conducted two independent experiments, where we studied the influence of Solanum nigrum unripe fruit extract on the toxicity of an organophosphorus insecticide fenitrothion. In the first variant of the experiment, Tenebrio molitor larvae were fed with blended fenitrothion (LC50) and the extract in four concentrations (0.01, 0.1, 1 and 10%) in ratio 1:1 for 3 days. In the second variant, a two-day application of fenitrothion (LC40) was preceded by a one-day extract treatment. The first variant did not show any increase in lethality compared to fenitrothion; however, ultrastructure observations exhibited swollen endoplasmic reticulum (ER) membranes in the midgut and nuclear and cellular membranes in the fat body, after application of blended fenitrothion and extract. An increased amount of heterochromatin in the fat body was observed, too. In the second variant, pre-treatment of the extract increased the lethality of larvae, decreased the level of glycogen and lipids in the fat body and disrupted integrity of midgut cellular membranes. S. nigrum extract, applied prior to fenitrothion treatment can be a factor increasing fenitrothion toxicity in T. molitor larvae. Thus, this strategy may lead to decreased emission of synthetic insecticides to the environment

The role of botanical treatments used in apiculture to control arthropod pests

Among pests of bees and beehives, arthropods make up a large and important group. Mites like Varroa destructor, Acarapis woodi, or Tropilaelaps spp., beetles (Aethina tumida, Oplostomus spp.), and lepidopterans (Galleria mellonella, Achroia grisella) decrease honey bee population and vitality, with subsequent significant colony production losses. Synthetic chemicals have been traditionally used to protect honey bee colonies from pests’ infestations but they have often been of poor selectivity, consequent high toxicity to bees and humans, and resistance development by the targeted apiary pests. The current European policy encourages the usage of eco-friendly methods to combat bee pests and the international research highlights plant secondary metabolites as candidate alternatives of significance. In this review, we argue the potential of plant-derived substances in the protection of the bee colonies against their arthropod pests. The before mentioned major apiary arthropods are briefly described followed by the recent reports on the botanical extracts and notable constituent compounds exhibiting activity against them. We discuss the different ways the essential oils are reported to be applied to the bee or the apiary, along with the importance of the application method to the exhibited efficacy. We designate synergism issues of blends, attractants, and repellency cases, as well as selectivity and mode of action as reported for bees or insect pests. © 2022, INRAE, DIB and Springer-Verlag France SAS, part of Springer Nature

Sublethal Effects of <i>Solanum nigrum</i> Fruit Extract and Its Pure Glycoalkaloids on the Physiology of <i>Tenebrio molitor</i> (Mealworm)

Background: Solanaceae plants produce glycoalkaloids (GAs) that affect various physiological processes of herbivorous insects and they are being tested as potential alternatives for synthetic pesticides. They cause lethal and sublethal effects. Nevertheless, their mode of action remains unclear. Therefore, we examined the effects of Solanum nigrum fruit extracts and pure glycoalkaloids on a model beetle, Tenebrio molitor. Methods: Plant extracts or pure alkaloids were added to the food of the larvae for three days. The lipid, glycogen, and protein content in the fat body and the midgut were determined, and the contractility of the heart, hindgut, and oviduct muscles was tested using the video-microscopy technique. Finally, the ultrastructure of the fat body and the midgut was observed using electron microscopy. Results: No lethal effects were noted. Sublethal changes were observed in the content of biomolecules, malformations of organelles, chromatin condensation, and heart and oviduct contractility. The observed effects differed between the tested glycoalkaloids and the extract. Conclusions: Both the extract and pure GAs have a wide range of effects that may result in impaired development, food intake, and reproduction. Some early effects may be used as bioindicators of stress. The effects of the extract and pure alkaloids suggest that the substances produced by the plant may act additively or synergistically

<i>Solanum nigrum</i> Fruit Extract Modulates Immune System Activity of Mealworm Beetle, <i>Tenebrio molitor</i> L.

Here, we report the first evidence concerning the modulation of insect immune system activity after applying Solanum nigrum fruit extract (EXT). We focused on two main issues: (1) is EXT cytotoxic for Tenebrio molitor haemocytes? and (2) how EXT affects the basic immune mechanisms of T. molitor. The results indicate cytotoxic action of 0.01 and 0.1% EXT on beetle haemocytes. Both the injection of EXT and incubating haemocytes with the EXT solution on microscopic slides significantly increased the number of apoptotic cells. However, 24 h after injection of 0.1% EXT cytotoxic effect of the tested extract probably was masked by the increased number of circulating haemocytes. Application of 0.01 and 0.1% EXT led to impairment of the activity of basic immune mechanisms such as phenoloxidase activity and the lysozyme-like antimicrobial activity of T. molitor haemolymph. Moreover, the EXT elicited significant changes in the expression level of selected immune genes. However, some of the immunomodulatory effects of EXT were different in beetles with and without an activated immune system. The obtained results are an essential step toward a complete understanding of the EXT mode of action on the T. molitor physiology and its potential usage in pest control

Differentiated Effects of Secondary Metabolites from <i>Solanaceae</i> and <i>Brassicaceae</i> Plant Families on the Heartbeat of <i>Tenebrio molitor</i> Pupae

The usage of insects as model organisms is becoming more and more common in toxicological, pharmacological, genetic and biomedical research. Insects, such as fruit flies (Drosophila melanogaster), locusts (Locusta migratoria), stick insects (Baculum extradentatum) or beetles (Tenebrio molitor) are used to assess the effect of different active compounds, as well as to analyse the background and course of certain diseases, including heart disorders. The goal of this study was to assess the influence of secondary metabolites extracted from Solanaceae and Brassicaceae plants: Potato (Solanum tuberosum), tomato (Solanum lycopersicum), black nightshade (Solanum nigrum) and horseradish (Armoracia rusticana), on T. molitor beetle heart contractility in comparison with pure alkaloids. During the in vivo bioassays, the plants glycoalkaloid extracts and pure substances were injected at the concentration 10&#8722;5 M into T. molitor pupa and evoked changes in heart activity. Pure glycoalkaloids caused mainly positive chronotropic effects, dependant on heart activity phase during a 24-h period of recording. Moreover, the substances affected the duration of the heart activity phases. Similarly, to the pure glycoalkaloids, the tested extracts also mainly accelerated the heart rhythm, however S. tuberosum and S. lycopersicum extracts slightly decreased the heart contractions frequency in the last 6 h of the recording. Cardioacceleratory activity of only S. lycopersicum extract was higher than single alkaloids whereas S. tubersoum and S. nigrum extracts were less active when compared to pure alkaloids. The most cardioactive substance was chaconine which strongly stimulated heart action during the whole recording after injection. A. rusticana extract which is composed mainly of glucosinolates did not significantly affect the heart contractions. Obtained results showed that glycoalkaloids were much more active than glucosinolates. However, the extracts depending on the plant species might be more or less active than pure substances