Pygidial Gland Secretion Research Articles

The morphology of the pygidial glands and the chemical composition of their secretions of four sphodrine ground beetle species (Carabidae: Platyninae)

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.

The pygidial gland secretion of Laemostenus punctatus (Coleoptera, Carabidae): a source of natural agents with antimicrobial, anti-adhesive, and anti-invasive activities

In the present study, we investigated in vitro the antimicrobial activity of the pygidial gland secretion of the guanophilic ground beetle Laemostenus (Pristonychus) punctatus (Dejean, 1828) and some of its chemicals against resistant and non-resistant bacteria and Candida species, the synergistic and additive potential of combinations of selected chemicals and antimicrobial drugs against resistant bacterial and fungal strains, anti-adhesive and anti-invasive potential of the secretion and formic acid alone and in selected combinations with antimicrobial drugs against methicillin-resistant Staphylococcus aureus (MRSA) toward spontaneously immortalized human keratinocyte cell line (HaCaT cells). In addition, we examined the antiproliferative activity of the secretion and formic acid in vitro. The tested secretion and the standards of formic and oleic acids possessed a significant level of antimicrobial potential against all tested strains (P < 0.05). The isolate from guano Pseudomonas monteilii showed the highest resistance to the secretion and formic acid, while MRSA achieved a significantly high level of susceptibility to all agents tested, particularly to the combinations of formic acid and antibiotics, but at the same time showed a certain level of resistance to the antibiotics tested individually. Candida albicans and C. tropicalis were found to be the most sensitive fungal strains to the secretion. Formic acid (MIC 0.0005 mg/mL) and gentamicin (MIC 0.0010 mg/mL) in the mixture achieved synergistic antibacterial activity against MRSA (FICI = 0.5, P < 0.05). The combination of formic acid, gentamicin and ampicillin accomplished an additive effect against this resistant bacterial strain (FICI = 1.5, P < 0.05). The secretion achieved a better inhibitory effect on the adhesion ability of MRSA toward HaCaT cells compared to formic acid alone, while formic acid showed better results regarding the invasion (P < 0.001). The combinations of gentamicin and ampicillin, as well as of formic acid and gentamicin and ampicillin achieved similar anti-adhesive and anti-invasive effects, with a slight advantage of formic acid and antibiotics in combination (P < 0.001). The secretion and formic acid were found to be non-toxic to HaCaT cells in vitro (IC50 ≥ 401 μg/mL).

The chemical composition of the secretions, their antibacterial activity, and the pygidial gland morphology of selected European Carabini ground beetles (Coleoptera: Carabidae)

The pygidial glands of carabids produce strong-smelling vapours. In this study, we examined the chemical composition of the gland secretions and the structure of the glands in five species of Carabini ground beetles (one species from the subtribe Calosomatina and four species from the subtribe Carabina): Calosoma (Calosoma) maderae (Fabricius, 1775), Carabus (Carabus) granulatus Linnaeus, 1758, C. (Limnocarabus) clathratus Linnaeus, 1760, C. (Carabus) ulrichii Germar, 1823, and C. (Procerus) gigas Creutzer, 1799. Additionally, we tested the antibacterial potential of the pygidial gland secretions of the two latter species against 11 bacterial strains. In order to detect the chemical content of the secretions, we used gas chromatography–mass spectrometry (GC–MS). The secretion extracts were applied against selected strains of medically important bacteria. We used bright-field microscopy to examine the morphology of the glands. We discovered a total of 11 chemical compounds in the pygidial gland extracts of the ground beetles we analysed. Ten of these compounds were identified as seven carboxylic acids, two hydrocarbons, and one aromatic aldehyde, while one chemical remained unidentified. Most of the components were isolated from the secretion of C. (L.) clathratus (nine), while the lowest number of compounds was found in C. (P.) gigas (two). Methacrylic acid was the most dominant compound by percentage in all five species, while angelic acid was also detected in all samples. As expected, salicylaldehyde was exclusively found in the species of the genus Calosoma Weber, 1801. The secretion of C. (P.) gigas was shown to achieve the highest level of antibacterial activity against Pseudomonas aeruginosa, Salmonella enterica, and S. typhimurium (even the same level as the positive control streptomycin), while the secretion of C. (C.) ulrichii achieved the highest antimicrobial potential against Staphylococcus epidermidis, S. aureus, Listeria monocytogenes, and Bacillus cereus. The most noticeable difference in the structure of the glands between the two genera is that the reservoir in Calosoma is more significantly narrowed as it leads into the efferent duct, compared to that of Carabus.

On the Diversity of Semiochemicals of the Pygidial Gland Secretions of Subterranean Ground Beetles (Coleoptera: Carabidae)

Pygidial glands are of great importance to ground beetles for defense against predators, especially for the species that live in subterranean habitats. The purpose of our study is to better understand the chemistry of the pygidial gland secretions of subterranean ground beetles, as well as the function and structure of the glands. We studied both the chemical composition of the pygidial gland secretion and morphology of the glands in adults of the troglophilic ground beetle species Laemostenus (Antisphodrus) cavicola (Schaum, 1858). The chemical composition of its defensive secretion was revealed using gas chromatography-mass spectrometry (GC-MS), while pygidial gland morphology of the beetle was investigated using bright-field microcopy. In total, seven chemical compounds were detected in the secretion mixture. Formic acid was the most dominant compound, followed by dodecyl acetate and undecane. Other chemicals were present in minor amounts. The morphological structure of the pygidial glands of L. (A.) cavicola was compared with the structure of the glands of the related congeneric troglophilic species Laemostenus (Pristonychus) punctatus (Dejean, 1828). Summary data on the semiochemicals that have been recorded so far in subterranean ground beetle species are presented, and the differences in the chemical composition of the secretions between and among troglobitic and troglophilic species are discussed. So far, forty-four compounds have been detected in four subterranean ground beetle species (two troglobites belonging to the tribe Trechini and two troglophiles belonging to the tribe Sphodrini). The results of this study indicate the great diversity of chemicals in the pygidial gland secretions of subterranean ground beetles.

Vinegaroon exposures reported to a Poison center

BackgroundVinegaroons (Mastigoproctus giganteus), also known as whip scorpions, are arachnids commonly found in the southwestern United States, parts of Mexico, and southern Florida. They do not bite, but have special teeth on the inside of the trochanters of the front appendages, used to crush prey. They are best known for having pygidial gland secretions containing 83% acetic acid, which are sprayed upon potential predators. There are no published descriptions of injury to human related to vinegaroon exposures. Our primary aim was to characterize types of exposures and clinical effects reported to a poison center that serves an area indigenous to this animal. MethodsThe database from a regional poison center was searched for all cases from 1998 to 2022 regarding human exposures to the vinegaroon. Data captured included age, sex, exposure route, type and duration of symptoms, and part of the body affected. ResultsThere were 50 exposures reported, with age range from 5 months to 54 years. Females represented 32 cases, males 17, and one unknown. Bites were more commonly described (36 cases), with 13 cases reported skin exposure to secretions, 3 ocular exposures, and 2 ingestions. Location of injury was upper extremities in 16 cases, lower in 11 cases, and torso in 3 cases. Symptoms were present in 88% and included pain, skin erythema, numbness or tingling, itching, and swelling. Ocular exposure were associated with pain in all 3 cases, with blurred vision in one case; effects lasted 1 h, 17 h, and more than 5 days. Four non-ocular exposures were followed to outcome, with duration of effects ranging from less than 1 h to more than 2 days. Discussion/conclusionIn a large series of vinegaroon exposures, females predominated, with most exposures occurring from skin contact with secretions. The most common symptoms were pain, erythema, numbness, itching, and swelling, which resolved in less than two days. Ocular exposures were associated with more symptoms and longer duration of effects.

Secretions of Pygidial Defensive Glands in Three Species of the Genus Bembidion (Carabidae), and Morphology of Pygidial Glands in B. (Peryphanes) dalmatinum

We studied the defensive pygidial gland secretions in the adults of three ground beetle species of the genus Bembidion. We detected 46 chemical compounds, of which we successfully identified 37, while nine remained unidentified. There were 4, 45 and 42 compounds in the pygidial gland secretions of Bembidion (Peryphanes) dalmatinum, B. (P.) deletum, and B. (Peryphus) subcostatum, respectively. n-Undecane, methacrylic acid and tiglic acid were common in all the studied species. Methacrylic acid was most abundant in the secretions of B. (P.) dalmatinum and B. (P.) deletum, while isovaleric acid in the secretion of B. (P.) subcostatum. We identified 30 compounds new to the tribe Bembidiini, 12 of which were recorded for the first time in the family Carabidae. In addition, we also analyzed the morphology of the pygidial glands in B. (P.) dalmatinum using both bright-field and nonlinear microscopy.

Pygidial glands of three ground beetle taxa (Insecta, Coleoptera, Carabidae): a study on their morphology and chemical composition of their secretions

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.

Chemistry and morphology of the pygidial glands in four Pterostichini ground beetle taxa (Coleoptera: Carabidae: Pterostichinae)

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.

Bat guano-dwelling microbes and antimicrobial properties of the pygidial gland secretion of a troglophilic ground beetle against them.

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 0.007mg/mL) and Escherichia fergusonii (MIC 0.028mg/mL). The most sensitive fungal isolates were M. guilliermondii, Penicillium expansum, and Trichoderma harzianum (MIC 0.15mg/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.

Pygidial gland secretions of Carabus Linnaeus, 1758 (Coleoptera: Carabidae): chemicals released by three species

It is a commonly known fact that all ground beetles possess abdominal pygidial glands with relatively similar gross structure and function among species. Still, morphology of the glands and composition of their secretions have not been studied in most ground beetle species. These exocrine glands and their products are mainly associated with defence in natural environments. In this paper, we studied three predatory ground beetle species of the genus Carabus Linnaeus, 1758, namely C. (Archicarabus) montivagus Palliardi, 1825, C. (Megodontus) caelatus Fabricius, 1801, and C. (M.) violaceus Linnaeus, 1758, to identify chemical components of their pygidial gland secretions. Altogether, 10 carboxylic acids were isolated from the analysed secretions [two from the secretion of C. (A.) montivagus, 10 from that of C. (M.) caelatus, and nine from that of C. (M.) violaceus]. The finding of 2-hexenoic acid in the secretion of C. (M.) caelatus is the first finding of it within the entire subfamily Carabinae. In addition, we also analysed the morphology of glands of the species C. (M.) violaceus.

Ant–hemipteran association decreases parasitism of Phenacoccus solenopsis by endoparasitoid Aenasius bambawalei

1. Mutualism between ants and honeydew‐excreting hemipterans is ubiquitous in the ecosystem. It is widely accepted that ant tending facilitates the colony growth of hemipterans by protecting them from predators and parasitoids. However, few studies have explored how ant tending helps defend against natural enemies.2. Ghost ant Tapinoma melanocephalum and the invasive mealybug Phenacoccus solenopsis have a close mutual relationship. Previous studies have shown that ghost ant tending can definitely reduce parasitism and visit frequency of Aenasius bambawalei, the dominant endoparasitoid of P. solenopsis. However, the ghost ant workers seldom attack the parasitoids. It is still unclear how the ghost ant adversely affects parasitoids. This study explored the mechanism underlying the impacts of ants on natural enemies of the mealybugs.3. Honeydew produced by P. solenopsis was an attractant to A. bambawalei. Parasitoids exhibited less searching activity, shorter longevity and lower parasitism when supplied with less honeydew. Aenasius bambawalei showed significant avoidance of pygidial gland secretions and visual cues of ghost ants. Parasitism in plants treated with 6‐methyl‐5‐hepten‐2‐one, actinidine, and gland extracts was significantly lower than that in plants treated only with solvents (paraffin oil or double‐distilled water).4. It is concluded that honeydew consumption by ghost ants could negatively influence the performance of parasitoids. The pygidial gland secretions and visual cues of ghost ants also significantly inhibit the parasitism. These results may contribute to a better understanding of the regulation mechanism in ant–hemipteran–enemy interactions.

Behavioral and chemical antipredator mechanisms in ground beetles: an attempt to reconcile within a phylogenetic frame

Like other coleopterans, Carabids are recognized for the use of passive and active strategies against their enemies. They are known to produce dangerous and distasteful chemicals and to use anti-predator behaviors such crypsis/mimicry, gregariousness, intra- and interspecific helpful interactions, sound production and aposematism. While defensive chemicals have been an important element in the successful diversification of ground beetles, in many tribes of this wide adephagan family many other systems have evolved and discovered. Here we present a review on the current knowledge of the avoidance mechanisms in carabids, with a discussion of data on chemical products, morphological adaptations and behaviors used against vertebrate and invertebrate predators. The most widespread strategy is discouragement of the predators using pygidial gland secretions, but passive anti-predator strategies are also displayed as group-protecting behavior through intraspecific aggregations or communal roosting. Most adult carabids tend to be dark in color and thus less detectable, but chemically well-protected taxa use the opposite strategy, displaying bright colors related to mimicry or aposematism. In aposematic species, group-protecting behavior has probably evolved from intraspecific gregariousness into multispecies aggregations. Larval behavioral strategies are related more closely to the epigean or underground lifestyle. In surface-living larvae, cannibalism avoidance and the usage of defensive substances have been reported for Chlaeniini. In this tribe we found an interesting coincidence between a high level of toxicity/distastefulness and a complex larval behavior. Based mainly on the genus-level supertree of Beutel et al. (2008) and the phylogenetic analyses of Ober & Maddison (2008) we reconstructed the evolutionary relationships among the chemical classes isolated inside the tribes. The scenario shows that several semiochemical classes (quinone/benzoquinones, formic acid, m-cresol, methacrylic acid, isovaleric acid, ß-necrodol, benzaldehyde, etc.) found in carabids are at least in part related to their life style and habitat selection. The proposed relationship still lacks other chemical data of the missing genera/species and even if the knowledge of carabid anti-predator strategies still seems to be in the pioneer stage, future studies on carabid anti-predator behavior look very promising.

Corpse management of the invasive Argentine ant inhibits growth of pathogenic fungi

A dead conspecific poses a potential pathogen risk for social animals. We have discovered that Argentine ants (Linepithema humile) prevent spread of pathogenic fungi from corpses by depositing the dead to combined toilet and refuse areas and applying pygidial gland secretion on them. The presence of a corpse in a nest increases this secretion behaviour. We identified three fungi growing on Argentine ant corpses. Growth of the Argentine ant pathogen Aspergillus nomius and the plant pathogen Fusarium solani on corpses was inhibited as long as the ants were constantly attending them as the ant anal secretion only delayed germination of their spores. In contrast, the effect of the ant anal secretion on the human pathogen Aspergillus fumigatus was much stronger: it prevented spore germination and, accordingly, the fungus no longer grew on the treated corpses. The Argentine ants are one of the world’s worst invasive alien species as they cause ecological and economical damage in their new habitats. Our discovery points at a novel method to limit Argentine ant colonies through their natural fungal pathogens.

Inhibition of tumour and non-tumour cell proliferation by pygidial gland secretions of four ground beetle species (Coleoptera: Carabidae)

Inhibition of the proliferation of human tumour cells and porcine non-tumour cells by the pygidial gland secretion released by adults of four ground beetle species was observed in this study. The sulphorhodamine B (SRB) assay was applied to establish the percentages of inhibition of the net growth of four human tumour cell lines and porcine liver primary non-tumour cells. The secretions of all tested ground beetle species were shown to have an antiproliferative effect on the tested cell lines. Special emphasis is put on the secretion of Abax parallelepipedus, which showed the highest antitumour potential and weakest inhibition of non-tumour cell proliferation. The antitumour and antiproliferative potential of the pygidial gland secretions of ground beetles is here demonstrated for the first time. It is suggested that certain organic acids are responsible for the action. Further investigation needs to be conducted in order to better understand the mechanisms governing the observed cytotoxic and antitumour activity.

Antifungal activity of the pygidial gland secretion of Laemostenus punctatus (Coleoptera: Carabidae) against cave-dwelling micromycetes.

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.

The pygidial gland secretion of the forest caterpillar hunter, Calosoma (Calosoma) sycophanta: the antimicrobial properties against human pathogens.

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.

Antimicrobial activity of the pygidial gland secretion of the troglophilic ground beetle Laemostenus (Pristonychus) punctatus (Dejean, 1828) (Insecta: Coleoptera: Carabidae).

The antimicrobial activity of the pygidial gland secretion released by adult individuals of the troglophilic ground beetle Laemostenus (Pristonychus) punctatus (Dejean, 1828), applying microdilution method with the aim to detect minimal inhibitory concentration, minimal bactericidal concentration and minimal fungicidal concentration, has been investigated. In addition, morphology of the pygidial glands is observed. 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 have showed antimicrobial properties against all strains of treated bacteria and fungi. Micrococcus flavus proved to be more resistant compared with other bacterial strains. More significant antimicrobial properties of the secretion are observed against Escherichia coli, which proved to be the most sensitive bacteria. Aspergillus fumigatus proved to be the most resistant, while Penicillium ochrochloron and Penicillium verrucosum var. cyclopium the most sensitive micromycetes. Commercial antibiotics Streptomycin and Ampicillin and antimycotics Ketoconazole and Bifonazole, applied as positive controls, showed higher antibacterial properties for all bacterial and fungal strains, except for P. ochrochloron, which proved to be more resistant on Ketoconazole compared with the pygidial gland secretion of L. (P.) punctatus. Apart from the role in ecological aspects, the antimicrobial properties of the tested secretion possibly might have medical significance in the future.

Antimicrobial activity of the pygidial gland secretion of three ground beetle species (Insecta: Coleoptera: Carabidae).

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.

Detection of a new piperideine alkaloid in the pygidial glands of some Stenus beetles.

Rove beetles of the genus Stenus produce and store bioactive alkaloids like stenusine (3), 3-(2-methylbut-1-enyl)pyridine (4), and cicindeloine (5) in their pygidial glands to protect themselves from predation and microorganismic infestation. The biosynthesis of stenusine (3), 3-(2-methylbut-1-enyl)pyridine (4), and cicindeloine (5) was previously investigated in Stenus bimaculatus, Stenus similis, and Stenus solutus, respectively. The piperideine alkaloid cicindeloine (5) occurs also as a major compound in the pygidial gland secretion of Stenus cicindeloides. The three metabolites follow the same biosynthetic pathway, where the N-heterocyclic ring is derived from L-lysine and the side chain from L-isoleucine. The different alkaloids are finally obtained by few modifications of shared precursor molecules, such as 2,3,4,5-tetrahydro-5-(2-methylbutylidene)pyridine (1). This piperideine alkaloid was synthesized and detected by GC/MS and GC at a chiral phase in the pygidial glands of Stenus similis, Stenus tarsalis, and Stenus cicindeloides.

Molecular Diversity of Compounds from Pygidial Gland Secretions of Cave-Dwelling Ground Beetles: The First Evidence.

Three adult cave-dwelling ground beetle species were induced to discharge secretions of their pygidial glands into vials. Dichloromethane extraction was used to obtain the secretions. In total, 42 compounds were identified by GC/MS analysis. Pheggomisetes ninae contained 32 glandular compounds, Laemostenus (Pristonychus) punctatus 13, whereas Duvalius (Paraduvalius) milutini had nine compounds. Caproic, oleic, palmitic, and stearic acids were present in the samples of all analyzed species. Undecane was predominant in the extract of L. punctatus. Palmitic acid was the major component in the secretion of D. milutini. Finally, the most abundant compounds in P. ninae secretion were heptacosene and nonacosadienes. Herein, we present the first data on the identification of pygidial gland secretion components in both troglophilous and troglobite cave-dwelling ground beetles. Some compounds are reported for the first time in the secretions of ground beetles and other higher or lower taxa. The adaptation to underground life has not led to a reduction or changes in the chemical defense mechanism in the analyzed troglophilous and troglobitic Platyninae and Trechinae taxa.