Cezary Tkaczuk

An infection event of the spittlebug Philaenus spumarius (Hemiptera, Aphrophoridae) has been described, for the first time, in northwest Italy. The causative agents were two entomopathogenic fungi belonging to Entomophthorales, specifically Zoophthora radicans and, more rarely, Batkoa major . The morphological description and molecular identification of fungi have been reported, in addition to recording meteorological data that may have affected the outbreak of the infection. When massive events are ongoing, entomopathogenic fungi really behave as determinant regulators of natural populations of arthropod pests and the possibility to stress their action in this direction should be deeply investigated.

The aim of the study was to identify the types of entomopathogenic fungi (EPF) and to determine the intensity of their occurrence in intensively cultivated agricultural soils in the Danubian Lowland, Slovakia. Soil samples were collected on two dates (spring, autumn) from seven sites where the following crops were grown: soybean, barley, alfalfa, sugar beet, maize, maize with the addition of a biostimulator and from wasteland. The entomopathogenic fungi from individual soil samples were isolated using the isolation method on a selective medium. Entomopathogenic fungi were identified microscopically based on the morphology of their microstructures and the morphology of colonies, using standard keys. The number of EPF was presented in CFU g–1 of dry matter of soil. Both in spring and autumn, EPF belonging to three genera were identified: Beauveria, Metarhizium and Cordyceps. Analyzing the average density of infectious units of the identified genera of EPF in both study dates on arable land, it was found that fungi of the genus Metarhizium spp. formed more infectious units in the spring study date, whereas fungi of the genera Beauveria spp. and Cordyceps spp. formed more infectious units in the autumn study date, and these differences were statistically significant.

The changing structure of modern cities intensifies anthropopressure, resulting in the need to create plans for the protection of biodiversity in cities. This can be achieved by establishing lawns and flower strips along the streets and maintaining parks and squares in cities, creating green infrastructure and contributing to sustainable urban development. However, this vegetation also requires protection that is safe for the environment and city residents. Entomopathogenic fungi (EPF) are among the most well-known and effective microorganisms that infect plant pests and conduct the disease process leading to their death. The aim of the study was to conduct a comparative analysis of the generic composition of EPF and determine the density of their colony-forming units (CFUs) in soils from flower strips and lawns located along the main communication routes of the city of Siedlce (Poland). Soil samples collected from two sites and two habitats (a flower strip and a lawn directly adjacent to it)—Site No. 1, Wyszyńskiego Street; Site No. 2, Jagiełły Street—in the spring and autumn of 2021/2022 and 2024. At each site within the habitat, three zones (repeats) were designated, spaced approximately 10–15 m apart. Approximately six samples were collected from each replication, and then a mixed sample was prepared. Four genera of EPF were found in the soil samples: Beauveria, Metarhizium, Cordyceps, and Akanthomyces. The location, habitat type, and season had a significant effect on the diversity of individual genera of fungi and the density of colony-forming units (CFUs) in the studied soils. The dominant types of EPF, forming the most CFUs in the soils from the studied flower strips and the adjacent lawns, were Metarhizium spp. and Beauveria spp. It was found that EPF occurred in higher densities in the soil from the studied habitats (flower strips and lawns) in autumn than in spring. Both of these semi-natural habitats constitute forms of urban greenery that increase biodiversity and provide valuable ecosystem services that support sustainable urban development.

Pyrethroids are chemical insecticides used on a large scale in agriculture, horticulture, and forest protection. In order to reduce their use in IPM, alternative methods of controlling insect pests are introduced, such as the use of biopesticides based on entomopathogenic fungi (EPF). Species of the Beauveria genus are characterized by a very broad spectrum of action, which is why they are often used to produce preparations based on EPF. The aim of the study was to determine the effect of different doses of tested pyrethroids on the colony growth and metabolic activity of EPF from the Beauveria genus. In vitro, the effect of three pyrethroids (deltamethrin, λ-cyhalothrin, and α-cypermethrin) added to SDA medium at a dose 10 times lower than the recommended field dose (A), the recommended field dose (B), and 10 times higher than the recommended field dose (C) on colony growth and metabolic activity of B. bassiana and B. brongniartii was tested. The research carried out showed that pyrethroid insecticides used in the experiment showed various toxic effects towards the tested EPF of the genus Beauveria. The studies conducted showed that on the 20th day of the observation, λ-cyhalothrin used in the recommended field dose limited the growth of B. bassiana to the least extent in relation to the other tested pyrethroids. However, with respect to the fungus B. brongniartii, no toxic effect of this pyrethroid was found. Based on the results obtained, it was found that λ-cyhalothrin used in the recommended field dose and 10 times lower than recommended significantly increased the metabolic activity of B. bassiana. In relation to the B. brongniartii strain, detlamethrin used in each of the tested concentrations significantly affected its viability.

The subphylum Entomophthoromycotina (Phylum Zoopagomycota) includes many arthropod pathogens, some of which are renowned for their abilities to alter host behavior prior to death and cause epizootics that impact host populations. The last checklist of arthropod-pathogenic species in this group was published in 1963 and consisted of 39 species in a single genus. Since then, more species have been named, and their taxonomy has changed extensively. We have constructed an updated checklist for species of Entomophthoromycotina in North America; this checklist includes species in the continental United States, Canada, and Mexico. Data were compiled based on available published literature and metadata available from the ARSEF culture collection, adjusting names based on current taxonomy. In North America, the arthropod-pathogenic Entomophthoromycotina now include 80 species belonging to 14 genera, within two classes, plus one species in a form genus. This checklist provides a current framework for future studies of the biodiversity of this group of fungi.

The experiment examined the effect of commercially available preparations containing humic substances in comparison with pure humic acids on the growth and germination of spores of entomopathogenic fungi (EPF) of the genus Beauveria in vitro. AmiAGRA, HumiAGRA, AlgoHUM (recommended field dose) and pure humic acids extracted from peat, brown coal and mushroom substrate were added to Sabouraud's culture substrate. Observation of the growth of colonies of the tested species of EPF was carried out every 5 days until day 20, measuring their diameter (mm). In the second stage of the experiment, the culture medium with the addition of preparations and pure humic acids was applied in a thin layer to the surface of glass slides and 0.1 ml of an aqueous solution with spores was introduced. Observation was carried out after 24 and 48 hours, and the results obtained were expressed as percentages of the control. The conducted research showed that on the 20th day of culture (on average), preparations containing humic substances had a stimulating effect, while pure acids slightly limited the growth of colonies of the tested isolates. The growth of the colony of the fungus B. basianna was most strongly stimulated by AmiAGRA, and B. brongniartii by HumiAGRA. The share of germinated spores of the tested isolates after 48 hours of contact with the substrate was higher than after 24 hours. It was found that more spores germinated on substrates with the addition of preparations containing humic substances than on pure humic acids.

The development of sustainable agricultural production involves the use of new plant protection products, with low toxicity to non-target organisms and high biodegradability. The aim of this study was to investigate the effect of commercially available preparations containing humic substances, in comparison with pure humic acids, on the growth and germination of spores of entomopathogenic fungi (EPFs) from the genus Metarhizium in vitro. AmiAGRA, HumiAGRA, AlgoHUM (recommended field dose) and pure humic acids extracted from peat, brown coal and spent mushroom substrate were added to Sabouraud’s culture substrate. Observation of the growth of the colonies of the tested species of EPFs was carried out every 5 days until day 20, measuring their diameter (mm). In the second stage of the experiment, the culture medium with the addition of preparations and pure humic acids was applied in a thin layer to the surface of glass slides and an aqueous solution with spores was introduced. Observation was carried out after 24 and 48 h, and the results obtained were expressed as percentages in relation to the control. The conducted research showed that on the 20th day of the culture (on average), preparations containing humic substances had a stimulating effect, while pure acids limited the growth of the colonies of the tested isolates slightly. The growth of M. anisopliae fungal colonies was most strongly stimulated by AlgoHUM, and M. flavoviride by HumiAGRA. The share of germinated spores after 48 h of contact with the substrate was higher than after 24 h, and more spores germinated on substrates with the addition of preparations containing humic substances than on pure humic acids.

The aim of the study was to determine the effect of zinc application in different doses with organic fertilization on the genera composition and the number of CFU (colony-forming units) of entomopathogenic fungi (EPF) in the soil. The experiment was carried out in greenhouse condi-tions. Soil samples for testing were collected in the third year of the experiment (spring, autumn), where the research objects were I – dose of zinc: control – lack (0) and 200, 400 and 600 mg Zn kg–1 of soil; II – no fertilization – 0 (CO), spent mushroom substrate (SMS), chicken (ChM) and cattle manure (CM). Entomopathogenic fungi were isolated using the method of isolation on a selective medium. In the course of the research, EPF of the genus: Beauveria, Metarhizium, Cordyceps and Lecanicillium were determined. The conducted research showed that entomopatho-genic fungi of the Metarhizium genera formed the most CFU in spring. In the autumn there were three times less of them. Statistical analysis showed that the number of CFUs of the identified genus of fungi (on average) in soil samples significantly depended on the dose of zinc applied, organic fertilization and the genus of fungus, but only for Metarhizium spp. The aim of the study was to determine the effect of zinc application in different doses with organic fertilization on the genera composition and the number of CFU (colony-forming units) of entomopathogenic fungi (EPF) in the soil. The experiment was carried out in greenhouse condi-tions. Soil samples for testing were collected in the third year of the experiment (spring, autumn), where the research objects were I – dose of zinc: control – lack (0) and 200, 400 and 600 mg Zn kg–1 of soil; II – no fertilization – 0 (CO), spent mushroom substrate (SMS), chicken (ChM) and cattle manure (CM). Entomopathogenic fungi were isolated using the method of isolation on a selective medium. In the course of the research, EPF of the genus: Beauveria, Metarhizium, Cordyceps and Lecanicillium were determined. The conducted research showed that entomopatho-genic fungi of the Metarhizium genera formed the most CFU in spring. In the autumn there were three times less of them. Statistical analysis showed that the number of CFUs of the identified genus of fungi (on average) in soil samples significantly depended on the dose of zinc applied, organic fertilization and the genus of fungus, but only for Metarhizium spp.

Cuticular lipids are essential for the functioning of all arthropods. They maintain water balance, are involved in intraspecific and interspecific chemical communication and prevent fungal and bacterial infections. Although, the insect's lipid layer is relatively well known, that of spiders is considerably less well known. Only a few species are studied in terms of the identification and composition of cuticular lipids. The results obtained so far are discussed in this article. Spiders are subjected to various forms of extraction and in the majority of cases the subsequent analysis using GC-MS identified hydrocarbons, carboxylic acids, esters, alcohols, aldehydes, ketones, methyl ethers and cholesterol. Among these lipids there are semiochemicals, mainly pheromones and kin recognition cues. Despite the few publications on spider cuticular lipids, this subject is significant in terms of searching for antimicrobial substances such as potential bio fungicides and medicines.

The aim of the study was to evaluate the potential of using five selected species of entomopathogenic fungi (Beauveria bassiana, B. brongniartii, Conidiobolus coronatus, Isaria fumosorosea, and Metarhizium robertsii) in the bioregulation of the dispersive stages of the parasitic nematode—Ascaris suum. Experimental cultures of each of the selected entomopathogenic fungi, as well as a control culture without fungi, were incubated with A. suum eggs at 26 °C for 28 days. Development of the A. suum eggs was observed using a light microscope on the 7th, 14th, 21st, and 28th days of incubation. The API-ZYM^® test was used to determine, semiquantitatively, the activity of 19 hydrolytic enzymes from the entomopathogenic fungi. The cytotoxicity of the fungi was determined using tetrazole salt MTT. It was found that none of the five tested strains of entomopathogenic fungi showed an ovicidal effect, and none of them colonized the A. suum egg shells. However, ovistatic activity was observed mainly until the 14th day of incubation by I. fumosorosea, M. robertsii, and B. bassiana. In the MTT test, M. robertsii showed moderate cytotoxicity, while the other species showed low cytotoxicity. Among the strains tested, I. fumosorosea showed the highest spectrum of hydrolase production (13 out of 19 enzymes gave a positive reaction from 3 to 5; 20–40 nM or more). The absence of morphological changes in the A. suum egg shells suggests that the antagonistic effect of the studied entomopathogenic fungi may be due to their cytotoxicity, associated with the production of secondary metabolites—toxins (M. robertsii) and enzymatic activity (I. fumosorosea).