Insecticidal activity of Bacillus thuringiensis towards Agrotis exclamationis larvae-A widespread and underestimated pest of the Palearctic zone.

acillus thuringiensis is an entomopathogenic bacterium commonly used as a bioinsecticide against numerous invertebrate pests. However, the efficacy of this microbe has not yet been determined towards Agrotis exclamationis-a lepidopteran, polyphagous pest, widespread throughout the Palearctic zone. In this work we have detected very low susceptibility of A. exclamationis to B. thuringiensis commercial strains, used as microbial formulations in pest control. To investigate this matter, the biological activity of six selected (Cry1Aa, Cry1Ca, Cry1Ia, Cry2Ab, Cry9Ea and Vip3Aa), heterogously-expressed Bacillus thuringiensis insecticidal proteins has been assessed towards A. exclamationis. Only Cry9Ea and Vip3Aa caused significant mortality in the tested pest species, with LC50 values of 950 and 140 ng/cm2, respectively. The histopathological effects of Cry9Ea and Vip3Aa on A. exclamationis were determined. On the other hand, Cry1- and Cry2-type toxins, which are the main active molecules of the majority of currently-used B. thuringiensis-based biocontrol agents (including the commercial strains tested in this work), did not cause mortality in target insect, but only different levels of growth inhibition. Moreover, in the case of Cry1Ca and Cry1Ia hormesis has been observed-a phenomenon that may be disadvantageous in implementation of these proteins in pest management. The obtained results broaden the existing knowledge regarding B. thuringiensis insecticidal protein target range and depict variable susceptibility of A. exclamationis to different groups of Cry/Vip toxins. This work indicates Cry9Ea and Vip3Aa as good candidates for efficient biological control of A. exclamationis and possibly other Agrotinae and discusses the potential use of Vip3-type and Cry9-type insecticidal proteins as successful bioinsecticides.

Different Dynamics of Reproductive Cell Development in Oviparous Clausilia bidentata and Ovoviviparous Ruthenica filograna Snails.

Most terrestrial snails are oviparous and only some are ovoviviparous. The physiology of the reproductive system and its activity in these two groups have received little attention. We compared the month-to-month dynamics of reproductive cell formation between two species of Clausiliidae snails: Ruthenica filograna (an ovoviviparous species) and Clausilia bidentata (an oviparous species). Our aim was to test the hypothesis that the seasonal activity of gametogenesis is determined by reproductive strategy (oviparity /ovoviviparity). The results showed that the frequency and number of reproductive cells in different stages varied month-to-month, within each species, and between the two species. Ruthenica filograna maintained its reproductive activity throughout the entire observation period, whereas the gonadal activity of C. bidentata changed with the seasons (it was temperature dependent). Our results are in tune with the hypothesis of Baur, that ovovivipary has some advantage over oviparity under harsher environmental conditions.

Reflectance spectroscopy and machine learning as a tool for the categorization of twin species based on the example of the Diachrysia genus.

In our work we used noninvasive point reflectance spectroscopy in the range from 400 to 2100 nm coupled with machine learning to study scales on the brown and golden iridescent areas on the dorsal side of the forewing of Diachrysia chrysitis and D. stenochrysis. We used our approach to distinguish between these species of moths. The basis for the study was a statistically significant collection of 95 specimens identified based on morphological feature and gathered during 23 years in Poland. The numerical part of an experiment included two independent discriminant analyses: stochastic and deterministic. The more sensitive stochastic approach achieved average compliance with the species identification made by entomologists at the level of 99-100%. It demonstrated high stability against the different configurations of training and validation sets, hence strong predictors of Diachrysia siblings distinctiveness. Both methods resulted in the same small set of relevant features, where minimal fully discriminating subsets of wavelengths were three for glass scales on the golden area and four for the brown. The differences between species in scales primarily concern their major components and ultrastructure. In melanin-absent glass scales, this is mainly chitin configuration, while in melanin-present brown scales, melanin reveals as an additional factor.

The TiO2-ZnO Systems with Multifunctional Applications in Photoactive Processes-Efficient Photocatalyst under UV-LED Light and Electrode Materials in DSSCs.

The main goal of the study was the hydrothermal-assisted synthesis of TiO2-ZnO systems and their subsequent use in photoactive processes. Additionally, an important objective was to propose a method for synthesizing TiO2-ZnO systems enabling the control of crystallinity and morphology through epitaxial growth of ZnO nanowires. Based on the results of X-ray diffraction analysis, in the case of materials containing a small addition of ZnO (≥5 wt.%), no crystalline phase of wurtzite was observed, proving that a high amount of modified titanium dioxide can inhibit the crystallization of ZnO. The transmission electron microscopy (TEM) results confirmed the formation of ZnO nanowires for systems containing ≥ 5% ZnO. Moreover, for the synthesized systems, there were no significant changes in the band gap energy. One of the primary purposes of this study was to test the TiO2-ZnO system in the photodegradation process of 4-chlorophenol using low-power UV-LED lamps. The results of photo-oxidation studies showed that the obtained binary systems exhibit good photodegradation and mineralization efficiency. Additionally, it was also pointed out that the dye-sensitized solar cells can be a second application for the synthesized TiO2-ZnO binary systems.

Hydrothermally Assisted Fabrication of TiO2-Fe3O4 Composite Materials and Their Antibacterial Activity.

The TiO2-Fe3O4 composite materials were fabricated via the hydrothermal-assisted technique. It was determined how the molar ratio of TiO2 to Fe3O4 influences the crystalline structure and morphology of the synthesized composite materials. The effect of the molar ratio of components on the antibacterial activity was also analyzed. On the basis of XRD patterns for the obtained titanium(IV) oxide-iron(II, III) oxide composites, the two separate crystalline forms-anatase and magnetite -were observed. Transmission electron microscopy revealed particles of cubic and tetragonal shape for TiO2 and spherical for Fe3O4. The results of low-temperature nitrogen sorption analysis indicated that an increase in the iron(II, III) oxide content leads to a decrease in the BET surface area. Moreover, the superparamagnetic properties of titanium(IV) oxide-iron(II, III) oxide composites should be noted. An important aim of the work was to determine the antibacterial activity of selected TiO2-Fe3O4 materials. For this purpose, two representative strains of bacteria, the Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus, were used. The titanium(IV) oxide-iron(II, III) oxide composites demonstrated a large zone of growth inhibition for both Gram-positive and Gram-negative bacteria. Moreover, it was found that the analyzed materials can be reused as antibacterial agents in three consecutive cycles with good results.

Hydrothermal-assisted synthesis of highly crystalline titania-copper oxide binary systems with enhanced antibacterial properties.

Binary oxide systems containing TiO2 and CuO were synthesized using hydrothermal treatment and shown to have enhanced antibacterial properties. A detailed investigation was made of the effect of the molar ratio of components (TiO2:CuO = 7:3, 5:5, 3:7, 1:9) on the physicochemical parameters and antibacterial activity. Analysis of morphology (SEM, TEM and HRTEM) confirmed the presence of spherical and sheet-shaped particles. On the XRD patterns for the binary oxide materials, two crystalline forms (anatase and monoclinic CuO) were observed. It was found that an increase in CuO content led to a decrease in the BET surface area of the TiO2-CuO binary oxide systems. The synthesized TiO2-CuO materials exhibited very good antibacterial activity against both Gram-positive (methicillin-resistant Staphylococcus aureus and Bacillus cereus) and Gram-negative (Salmonella Enteritidis and Pseudomonas aeruginosa) bacteria. The obtained results show that TiO2-CuO oxide materials may have applications in the biomedical and food industries.

The effects of structurally different siderophores on the organelles of Pinus sylvestris root cells.

MAIN CONCLUSION: Siderophores are a driver of Pinus sylvestris root responses to metabolites secreted by pathogenic and mycorrhizal fungi. Structurally different siderophores regulate the uptake of Fe by microorganisms and may play a key role in the colonization of plants by beneficial or pathogenic fungi. Siderophore action, however, may be dependent on the distribution of Fe within cells. Here, the involvement of siderophores in determining the changes of organelle morphology and element composition of some cellular fractions of root cells in Pinus sylvestris to trophically diverse fungi was investigated. Changes in the morphology and concentrations of different elements within organelles of root cells in response to three structurally different siderophores were examined by transmission electron microscopy combined with energy-dispersive X-ray spectroscopy. Weak development of mitochondrial cristae and the deposition of backup materials in plastids occurred in the absence of Fe in the structures of triacetylfusarinine C and ferricrocin. In response to metabolites of both pathogenic and mycorrhizal fungi, Fe accumulated mainly in the cell walls and cytoplasm. Fe counts increased in all of the analyzed organelles in response to applications of ferricrocin and triacetylfusarinine C. Chelation of Fe within the structure of siderophores prevents the binding of exogenous Fe, decreasing the abundance of Fe in the cell wall and cytoplasm. The concentrations of N, P, K, Ca, Mn, Cu, Mg, and Zn also increased in cells after applications of ferricrocin and triacetylfusarinine C, while the levels of these elements decreased in the cell wall and cytoplasm when Fe was present within the structure of the siderophores. These results provide insight into the siderophore-driven response of plants to various symbionts.

The natural insect peptide Neb-colloostatin induces ovarian atresia and apoptosis in the mealworm Tenebrio molitor.

BACKGROUND: The injection of Neb-colloostatin into T. molitor females causes gonadoinhibitory effects on ovarian development. This peptide inhibits intercellular space formation (patency) in follicular epithelium and results in slowed vitellogenesis, delayed ovulation, reduced number of eggs laid and presumably cell death in the terminal follicles. However, as does the form of cell death in the terminal follicle, the mode of action of Neb-colloostatin remains unknown. RESULTS: We tested Neb-colloostatin for a sterilizing effect on females of Tenebrio molitor. We report that injection of nanomolar doses of Neb-colloostatin induce ovarian follicle atresia in 4-day old females during their first gonadotropic cycle. Light microscope observations revealed morphological changes in the ovary: after Neb-colloostatin injection the terminal oocytes are significantly smaller and elicit massive follicle resorption, but the control terminal follicles possess translucent ooplasm in oocytes at different stages of vitellogenesis. A patency is visible in follicular epithelium of the control vitellogenic oocytes, whereas peptide injection inhibits intercellular space formation and, in consequence, inhibits vitellogenesis. Confocal and electron microscope examination showed that peptide injection causes changes in the morphology indicating death of follicular cells. We observed F-actin cytoskeleton disorganization, induction of caspase activity, changes in chromatin organization and autophagic vacuole formation. Moreover, the apical cytoplasm of follicular cells is filled with numerous free ribosomes, probably indicating a higher demand for protein biosynthesis, especially in preparation for autophagic vacuole formation. On the other hand, the process of polyribosomes formation is inhibited, indicating the contributing effect of this hormone. CONCLUSION: Neb-colloostatin induces atresia in the mealworm ovary. Degeneration of T. molitor follicles includes changes in morphology and viability of follicular cells, and oosorption as a consequence of these changes.