RESUMO
Ground beetles possess a pair of pygidial glands that produce and release secretions that play an important role in defense against predators. The morphology of these glands and the chemical composition of their products were studied in four species of the tribe Sphodrini: Calathus (Calathus) fuscipes (Goeze, 1777), C. (Neocalathus) cinctus Motschulsky, 1850, C. (N.) melanocephalus (Linnaeus, 1758) and Laemostenus (Antisphodrus) elongatus (Dejean, 1828). The morphological analyzes of the glands of the four taxa mentioned were carried out for the first time using bright-field and nonlinear microscopy. All morphological structures were precisely measured and photographed. The pygidial gland secretions of C. (C.) fuscipes and L. (A.) elongatus were analyzed for the first time using gas chromatography-mass spectrometry. A total of 30 compounds were detected from the extracts of pygidial gland secretions of the four Sphodrini species studied. The simplest chemical mixture was found in L. (A.) elongatus, while the most complex secretion was that of C. (C.) fuscipes. 1-Undecanol, which we were able to detect in all taxa examined here, and dodecyl butyrate, which was detected in the three Calathus species, have never before been detected in the secretions of ground beetles.
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In this paper, the genus Vitrea Fitzinger, 1833 in Serbia is reviewed. All previous literature data on this genus from Serbia are summarised and used to discuss its distribution in the country and create distribution maps, supplemented by new material collected by the authors. All Serbian species are figured. For each species, a brief description of the examined specimens, data on previous findings in Serbia, the material (including types) that were analysed, the distribution and habitats in Serbia they inhabit, as well as remarks on specific species are given. A new species, Vitreavirgo Gojsina & Dedov, sp. nov., is described from a pit on Mt. Devica in eastern Serbia. Vitreapygmaea (O. Boettger, 1880) is reported for the first time for the territory of Serbia. As some Vitrea species have a narrow geographical range and prefer certain habitats, they are particularly vulnerable to habitat changes, which is also discussed in the paper. An identification key for all hitherto known Serbian species is given.
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Pygidial glands are a common feature of all adephagans and their products play an important role in defense against predators. The morphology of the pygidial glands and the chemistry of their secretion were studied for the first time in two species of tiger beetles - Cicindela (Cicindela) sylvicola Dejean, 1822 and Cylindera (Cylindera) germanica (Linnaeus, 1758). The glands were examined by both bright-field microscopy and nonlinear microscopy. All morphological structures of the glands were measured and described in detail. The structures mentioned were compared with those of related taxa. The secretion extracts from the pygidial glands of the investigated taxa contained a total of 24 compounds, which were detected by gas chromatography-mass spectrometry (GC-MS). The secretion mixture of C. (C.) germanica was more complex (21 chemicals) than that of C. (C.) sylvicola (11 ones). Benzaldehyde was present in both secretion samples. Hydrocarbons were the most abundant group of secretory compounds. The purpose of the compounds, their distribution within the subfamily Cicindelinae and their effects on the ecology of the group were discussed.
Assuntos
Besouros , Animais , Besouros/anatomia & histologia , MicroscopiaRESUMO
Pygidial gland secretions are used as repellent defensive allomones in ground beetles. We provide the first precise data on the chemical composition and antimicrobial potency of the secretion of the blue ground beetle, as well as on the morphology of its pygidial glands. The latter structures were not previously studied chemoecologically and morphologically, and we hypothesized that their secretion may have some antimicrobial action, as is the case with certain Carabus species. Gas chromatography-mass spectrometry (GC-MS) was used to identify methacrylic and angelic acids as dominant chemicals in the secretion from individuals of three populations of the blue ground beetle in Serbia. We tested its secretion against selected strains of medically important microorganisms. The secretion exibits antimicrobial action against certain bacterial species and all tested micromycetes. The most significant antifungal effect of the secretion was against Penicillium ochrochloron, which is more sensitive to the secretion than to commercial antifungal drugs ketoconazole and bifonazole. Bifonazole achieved minimum inhibitory concentrations against Trichoderma viride at more than three times higher value than did the secretion, indicating a significant antifungal effect of the secretion against this micromycete as well. Additionally, we tested commercially available standards of two dominant chemicals in the secretion to investigate their interaction and antimicrobial role in the secretion. Finally, we describe all glandular morpho-functional units of the blue ground beetle. Our results suggest that the secretion of the blue ground beetle may serve not only defensive but also antimicrobial functions, which likely aid the survival of this beetle in the microbial-rich forest litter habitat.
Assuntos
Anti-Infecciosos , Besouros , Animais , Anti-Infecciosos/farmacologia , Bactérias , Secreções Corporais/química , Besouros/química , Testes de Sensibilidade MicrobianaRESUMO
Morphology of the pygidial glands and chemical compositions of their secretion were analysed in the adults of three selected ground beetle taxa. Secretions of pygidial glands of Cychrus (Cychrus) semigranosus, Patrobus atrorufus and Pterostichus (Platysma) niger were chemically tested. Additionally, pygidial glands of the latter two species were investigated using bright-field microscopy and nonlinear microscopy and morphological features of the glands were described in detail. Both C. (C.) semigranosus and P. atrorufus were studied for the first time in terms of chemical ecology, while the latter species was analysed for the first time in terms of pygidial gland morphology. Altogether, eight compounds were detected in the dichloromethane extracts of the pygidial gland secretions of the three ground beetle taxa analysed. The simplest secretion mixtures were present in C. (C.) semigranosus and P. atrorufus (with two compounds each), while the extract of P. (P.) niger contained five compounds. The presence of 1-tetradecanol in the secretion of P. (P.) niger represents the first finding of this compound from the pygidial gland secretion extracts of ground beetles.
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Estruturas Animais/fisiologia , Secreções Corporais/química , Besouros/fisiologia , Animais , Besouros/classificação , Feminino , Masculino , Especificidade da EspécieRESUMO
Morphology of the pygidial glands and chemical composition of their secretions in adults of four ground beetle representatives of the Pterostichini tribe (Coleoptera: Carabidae) were analysed. Molops (Stenochoromus) montenegrinus, Pterostichus (Cophosus) cylindricus, P. (Feronidius) melas and P. (Pseudomaseus) nigrita were chemically tested, while the latter three species were morphologically investigated. Pterostichus (C.) cylindricus, P. (P.) nigrita and M. (S.) montenegrinus were chemically studied for the first time. Altogether, 23 chemical compounds were isolated using gas chromatography-mass spectrometry (GC-MS), of which some are new for Pterostichini or even Carabidae. Methacrylic acid was present in all species analysed. It was predominant in the secretion extract of P. (C.) cylindricus and P. (F.) melas. Isobutyric and 2-methylbutyric acids were the major components in the secretion of M. (S.) montenegrinus. Undecane, methacrylic and tiglic acids were the main components in the secretion of P. (P.) nigrita. The simplest chemical mixture was found in P. (C.) cylindricus (two compounds), while the most complex one was detected in P. (P.) nigrita (15 compounds). No significant differences in the chemical composition of the pygidial gland secretions were evidenced in P. (C.) cylindricus sampled from the same area and in the same season in two different years. Morphology of the pygidial glands of the studied species was analysed for the first time. Morphological features of the pygidial glands were observed using bright-field microscopy and nonlinear microscopy and described in details.
Assuntos
Secreções Corporais/química , Besouros/anatomia & histologia , Glândulas Exócrinas/anatomia & histologia , Animais , Besouros/fisiologia , Glândulas Exócrinas/fisiologia , Especificidade da EspécieRESUMO
Bat guano is an important source of microbial diversity in caves and can be a source of potential pathogens. Laemostenus (Pristonychus) punctatus is a guanophilic ground beetle species, which pygidial gland secretion exhibits action against pathogenic and other microbes. The distribution and diversity of microbes in bat guano from a karstic cave were determined in this study. Additionally, antimicrobial activity of the pygidial gland secretion of L. (P.) punctatus against guano-dwelling microbes was tested; minimal inhibitory concentration (MIC) and chemical composition of the secretion were analyzed. In total, 63 different bacterial species and 16 fungal morphotypes were isolated from guano samples by the cultivation method and confirmed using and phenotypic characterization and molecular identification. There was a difference in the composition of certain microorganisms between the sampling points (cave locations) and between the guano layers. The largest number of bacterial isolates belongs to the genera Lysinibacillus and Paenibacillus, while Pseudomonas species were highly abundant at the innermost sampling point. For the guanophilic fungi, the majority are ascomycetes, with Penicillium and Aspergillus as the most dominant genera. Meyerozyma guilliermondii was the only yeast species found in the guano samples. The most sensitive isolates were Enterococcus eurekensis (MIC 0.007 mg/mL) and Escherichia fergusonii (MIC 0.028 mg/mL). The most sensitive fungal isolates were M. guilliermondii, Penicillium expansum, and Trichoderma harzianum (MIC 0.15 mg/mL). This study opens a new possibility for better understanding of ecological relations between microorganisms and troglophilic ground beetles and for detailed investigations of morpho-anatomical aspects of pygidial glands.
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Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Quirópteros/microbiologia , Besouros/química , Glândulas Exócrinas/química , Fezes/microbiologia , Fungos/efeitos dos fármacos , Animais , Bactérias/classificação , Secreções Corporais/química , Cavernas/microbiologia , Besouros/anatomia & histologia , Feminino , Fungos/classificação , Masculino , SérviaRESUMO
Macroecologists seek to identify drivers of community turnover (ß-diversity) through broad spatial scales. However, the influence of local habitat features in driving broad-scale ß-diversity patterns remains largely untested, owing to the objective challenges of associating local-scale variables to continental-framed datasets. We examined the relative contribution of local- versus broad-scale drivers of continental ß-diversity patterns, using a uniquely suited dataset of cave-dwelling spider communities across Europe (35-70° latitude). Generalized dissimilarity modelling showed that geographical distance, mean annual temperature and size of the karst area in which caves occurred drove most of ß-diversity, with differential contributions of each factor according to the level of subterranean specialization. Highly specialized communities were mostly influenced by geographical distance, while less specialized communities were mostly driven by mean annual temperature. Conversely, local-scale habitat features turned out to be meaningless predictors of community change, which emphasizes the idea of caves as the human accessible fraction of the extended network of fissures that more properly represents the elective habitat of the subterranean fauna. To the extent that the effect of local features turned to be inconspicuous, caves emerge as experimental model systems in which to study broad biological patterns without the confounding effect of local habitat features.
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Meio Ambiente , Aranhas/fisiologia , Animais , Biodiversidade , Ecossistema , Europa (Continente) , Geografia , Especificidade da Espécie , TemperaturaRESUMO
BACKGROUND: Spiders (Arachnida: Araneae) are widespread in subterranean ecosystems worldwide and represent an important component of subterranean trophic webs. Yet, global-scale diversity patterns of subterranean spiders are still mostly unknown. In the frame of the CAWEB project, a European joint network of cave arachnologists, we collected data on cave-dwelling spider communities across Europe in order to explore their continental diversity patterns. Two main datasets were compiled: one listing all subterranean spider species recorded in numerous subterranean localities across Europe and another with high resolution data about the subterranean habitat in which they were collected. From these two datasets, we further generated a third dataset with individual geo-referenced occurrence records for all these species. NEW INFORMATION: Data from 475 geo-referenced subterranean localities (caves, mines and other artificial subterranean sites, interstitial habitats) are herein made available. For each subterranean locality, information about the composition of the spider community is provided, along with local geomorphological and habitat features. Altogether, these communities account for > 300 unique taxonomic entities and 2,091 unique geo-referenced occurrence records, that are made available via the Global Biodiversity Information Facility (GBIF) (Mammola and Cardoso 2019). This dataset is unique in that it covers both a large geographic extent (from 35° south to 67° north) and contains high-resolution local data on geomorphological and habitat features. Given that this kind of high-resolution data are rarely associated with broad-scale datasets used in macroecology, this dataset has high potential for helping researchers in tackling a range of biogeographical and macroecological questions, not necessarily uniquely related to arachnology or subterranean biology.
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Modern document protection relies on the simultaneous combination of many optical features with micron and submicron structures, whose complexity is the main obstacle for unauthorized copying. In that sense, documents are best protected by the diffractive optical elements generated lithographically and mass-produced by embossing. The problem is that the resulting security elements are identical, facilitating mass-production of both original and counterfeited documents. Here, we prove that each butterfly wing-scale is structurally and optically unique and can be used as an inimitable optical memory tag and applied for document security. Wing-scales, exhibiting angular variability of their color, were laser-cut and bleached to imprint cryptographic information of an authorized issuer. The resulting optical memory tag is extremely durable, as verified by several century-old insect specimens still retaining their coloration. The described technique is simple, amenable to mass-production, low cost and easy to integrate within the existing security infrastructure.
Assuntos
Biomimética , Borboletas/anatomia & histologia , Documentação , Fenômenos Ópticos , Segurança , Asas de Animais , AnimaisRESUMO
We investigated morphometric variation in size and shape of the head, pronotum, and abdomen between the taxa and sexes of two ground beetles, Carabus coriaceus cerisyi Dejean, 1826 and C. kollari praecellens Palliardi, 1825. These two taxa differ in overall size, and both of them are characterized by significant sexual size dimorphism. In many taxa, allometry, the relationship between changes in shape and changes in size, can be a major component of intra- and interspecific variation in body shape. In the present study, we applied landmark-based geometric morphometrics to explore allometric and non-allometric components of shape variation between the taxa and more importantly between sexes within the taxa. We were able to show that the differences of shape between the taxa cannot be explained by allometric changes, as allometry explains only a small amount of total shape variation between the taxa, which was expected due to the fact that the taxa belong to separate subgenera. Between the sexes, on the other hand, allometry contributes largely to the variation, particularly in abdomen shape. However, the differences of head and pronotum shape between the sexes cannot be entirely explained in terms of allometric scaling. Our results indicate that allometry contributes largely to differences of body shape between the sexes, while differences between the taxa are influenced by other factors and processes.
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Besouros/anatomia & histologia , Animais , Tamanho Corporal , Feminino , Masculino , Caracteres SexuaisRESUMO
Leptomeson Jeannel, 1924, originally treated as a subgenus of Anthroherpon Reitter, 1889 (Jeannel, 1924), was erected to a distinct genus by Guéorguiev (1990). It currently includes 13 endemic taxa (nine species and four subspecies) (Perreau, 2015), of which five species are recently described (Giachino et al., 2011). All Leptomeson taxa are troglobitic, montane or insular, and are distributed in a narrow Dinaric area in the proximity to the Adriatic Sea coast belonging to Croatia and Bosnia and Herzegovina (Perreau, 2000; Giachino et al., 2011) (Fig. 1).
Assuntos
Besouros , Animais , CroáciaRESUMO
Chaetomargoreicheia Magrini Bulirsch, 2005 is a recently established genus of scaritine ground beetles (Bulirsch Guéorguiev, 2008) (treated by certain authors as a subgenus of the genus Reicheadella Reitter, 1913) (Magrini Bulirsch, 2005; Balkenohl, 2017) which currently contains two endogean species inhabiting the Balkan Peninsula: Chaetomargoreicheia zoufali (Reitter, 1913) and C. lakotai (Magrini Bulirsch, 2005) (Balkenohl, 2003, 2017; Magrini Bulirsch, 2005; Bulirsch Guéorguiev, 2008). The aforementioned species are montane and inhabit confined geographic areas (Jeannel, 1957; Magrini Bulirsch, 2005). C. zoufali was found only in the surroundings of the village of Ravno (collected beneath a deep layer of leaf-litter), Mt. Bjelasnica, near Trebinje, E Herzegovina (Bosnia and Herzegovina), while C. lakotai was found at the entrance of a cave nearby a road (collected under a huge stone), in Mt. Lovcen, near Kotor, S Montenegro (Reitter, 1913; Holdhaus, 1924; Jeannel, 1957; Magrini Bulirsch, 2005). Chaetomargoreicheia species represent quite rare, stenoendemic taxa, for each of the up-to-now known species only one specimen has been collected by hand so far (Reitter, 1913; Jeannel, 1957; Magrini Bulirsch, 2005). The fourth author of the current study investigated numerous underground and endogean high-altitude habitats in Dalmatia (Croatia) in the last few years. As a result of the exploration, he has recently collected a small sample of scaritine ground beetles from a cave in S Croatia. After thorough analysis of the sample, we have identified a new Chaetomargoreicheia species.
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Besouros , Animais , Península Balcânica , Bósnia e Herzegóvina , Croácia , Masculino , MontenegroRESUMO
The trechine ground beetle taxa, Velesaphaenops gen. n., Velesaphaenops tarensis sp. n., and Acheroniotes lethensis sp. n., are described and diagnosed. The new taxa differ clearly from their closest relatives in a number of external characters and the shape of the genitalia. They probably belong to phyletic lineages of Pliocene age (the age of the palaeokarst of Kamena Gora and Mt. Tara). The new taxa are endemic to western and southwestern Serbia. Keys to the aphaenopsoid trechine genera in Serbia and to species of the genus Acheroniotes Lohaj Lakota, 2010 are appended.
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Besouros , Filogenia , Animais , Genitália , SérviaRESUMO
Seven new troglobitic leiodid beetle taxa (six new species and one new subspecies) from several caves and pits in the Balkan Peninsula (Bosnia and Herzegovina and Serbia) are described and diagnosed. Three new species belong to the genus Adelopidius Apfelbeck, 1907, two-to the genus Pholeuonopsis Apfelbeck, 1901, one-to the genus Leonhardia Reitter, 1901, while a new subspecies belongs to the genus Apholeuonus Reitter, 1889. All important morphological characteristics of the new taxa have been mentioned and imaged, including the aspects of the male and female genitalia. The new leiodid taxa are properly distinguished from their relatives and are endemics of the Dinaric mountain chain. A key to Adelopidius taxa is added.
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Distribuição Animal , Besouros , Animais , Península Balcânica , Bósnia e Herzegóvina , Feminino , Masculino , SérviaRESUMO
The insect cuticle serves a multitude of purposes, including: mechanical and thermal protection, water-repelling, acoustic signal absorption and coloration. The influence of cuticular structures on infrared radiation exchange and thermal balance is still largely unexplored. Here we report on the micro- and nanostructured setae covering the elytra of the longicorn beetle Rosalia alpina (Linnaeus, 1758) (Coleoptera: Cerambycidae) that help the insect to survive in hot, summer environments. In the visible part of the spectrum, scale-like setae, covering the black patches of the elytra, efficiently absorb light due to the radiation trap effect. In the infrared part of the spectrum, setae of the whole elytra significantly contribute to the radiative heat exchange. From the biological point of view, insect elytra facilitate camouflage, enable rapid heating to the optimum body temperature and prevent overheating by emitting excess thermal energy.
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Exoesqueleto/química , Regulação da Temperatura Corporal , Besouros/anatomia & histologia , Animais , Temperatura Alta , Luz , Modelos Biológicos , TermografiaRESUMO
The antifungal potential of the pygidial gland secretion of the troglophilic ground beetle Laemostenus punctatus from a cave in Southeastern Serbia against cave-dwelling micromycetes, isolated from the same habitat, has been investigated. Eleven collected samples were analyzed and 32 isolates of cave-dwelling fungi were documented. A total of 14 fungal species were identified as members of the genera Aspergillus, Penicillium, Alternaria, Cladosporium, Rhizopus, Trichoderma, Arthrinium, Aureobasidium, Epicoccum, Talaromyces, and Fusarium. Five isolates were selected for testing the antifungal activity of the pygidial gland secretion: Talaromyces duclauxi, Aspergillus brunneouniseriatus, Penicillium sp., Rhizopus stolonifer, and Trichoderma viride. The microdilution method has been applied to detect minimal inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs). The most sensitive isolate was Penicillium sp., while the other isolates demonstrated a high level of resistance to the tested agent. L. punctatus has developed a special mechanism of producing specific compounds that act synergistically within the secretion mixture, which are responsible for the antifungal action against pathogens from the cave. The results open opportunities for further research in the field of ground beetle defense against pathogens, which could have an important application in human medicine, in addition to the environmental impact, primarily.
Assuntos
Antifúngicos/farmacologia , Besouros/química , Fungos/efeitos dos fármacos , Animais , Antifúngicos/isolamento & purificação , Secreções Corporais/química , Secreções Corporais/microbiologia , Cavernas , Besouros/microbiologia , Testes de Sensibilidade Microbiana , SérviaRESUMO
Here we report how interference and scattering-enhanced absorption act together to produce the golden wing patches of the burnished brass moth. The key mechanism is scattering on rough internal surfaces of the wing scales, accompanied by a large increase of absorption in the UV-blue spectral range. Unscattered light interferes and efficiently reflects from the multilayer composed of the scales and the wing membranes. The resulting spectrum is remarkably similar to the spectrum of metallic gold. Subwavelength morphology and spectral and absorptive properties of the wings are described. Theories of subwavelength surface scattering and local intensity enhancement are used to quantitatively explain the observed reflectance spectrum.
Assuntos
Cor , Mariposas , Asas de Animais , Animais , Microscopia , Microscopia Eletrônica de Varredura , Modelos Biológicos , Mariposas/anatomia & histologia , Mariposas/efeitos da radiação , Espalhamento de Radiação , Asas de Animais/anatomia & histologia , Asas de Animais/efeitos da radiaçãoRESUMO
Recently, various insect secretions have been tested as possible antimicrobial agents. In beetles, these secretions are essentially products of various exocrine glands, of which particular emphasis is on pygidial glands that are common for the suborder Adephaga. The antimicrobial potential of the pygidial gland secretion isolated from the adults of Calosoma sycophanta against human pathogens has been tested and compared with the potential of other carabid species, particularly within the tribe Carabini. The antimicrobial assay includes a microdilution method which was applied in order to determine the minimal inhibitory, minimal fungicidal and minimal bactericidal concentrations. It has been tested the effect of the secretion against eight strains of fungal and eight strains of bacterial species. The secretion sample of the tested carabid species has shown statistically significant antifungal effect against all strains of treated micromycetes, the highest in comparison with previously tested carabids (Carabus spp., Laemostenus punctatus and Abax parallelepipedus). Aspergillus versicolor proved to be the most sensitive micromycete, while the remaining seven fungal strains have shown the same level of sensibility. In comparison with commercial mycotics ketoconazole and bifonazole, applied as positive controls, the tested secretion showed much higher antifungal activity for all fungal strains. Antibacterial effect has been manifested only against one bacterial strain (Escherichia coli), contrary to other previously studied carabid species. These observations might have a significant impact on the ecological domain and possible purpose in biomedical studies and applications in the future. Additionally, morpho-histology of the pygidial glands of C. sycophanta is investigated and discussed.
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Bactérias/efeitos dos fármacos , Besouros/metabolismo , Fungos/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/metabolismo , Antifúngicos/isolamento & purificação , Antifúngicos/metabolismo , Aspergillus/efeitos dos fármacos , Besouros/anatomia & histologia , Escherichia coli/efeitos dos fármacos , Glândulas Exócrinas/metabolismo , Glândulas Exócrinas/ultraestrutura , Humanos , Testes de Sensibilidade MicrobianaRESUMO
The antimicrobial properties of the pygidial gland secretions released by the adults of the three ground beetle species, Carabus ullrichii, C. coriaceus, and Abax parallelepipedus, have been tested. Microdilution method was applied for detection of minimal inhibitory concentrations (MICs), minimal bactericidal concentrations (MBCs), and minimal fungicidal concentrations (MFCs). Additionally, morpho-histology of the pygidial glands is investigated. We have tested 16 laboratory and clinical strains of human pathogens--eight bacterial both gram-positive and gram-negative species and eight fungal species. The pygidial secretion samples of C. ullrichii have showed the strongest antimicrobial effect against all strains of treated bacteria and fungi. Staphylococcus aureus, Lysteria monocytogenes, and Salmonella typhimurium proved to be the most sensitive bacterial strains. Penicillium funiculosum proved to be the most sensitive micromycete, while P. ochrochloron and P. verrucosum var. cyclopium the most resistant micromycetes. The pygidial secretion of C. coriaceus has showed antibacterial potential solely against Pseudomonas aeruginosa and antifungal activity against Aspergillus fumigatus, A. versicolor, A. ochraceus, and P. ochrochloron. Antibacterial properties of pygidial gland secretion of A. parallelepipedus were achieved against P. aeruginosa, while antifungal activity was detected against five of the eight tested micromycetes (A. fumigatus, A. versicolor, A. ochraceus, Trichoderma viride, and P. verrucosum var. cyclopium). Commercial antibiotics Streptomycin and Ampicillin and mycotics Ketoconazole and Bifonazole, applied as the positive controls, showed higher antibacterial/antifungal properties for all bacterial and fungal strains. The results of this observation might have a significant impact on the environmental aspects and possible medical purpose in the future.