Kalotermes Flavicollis Research Articles

Attacks of Kalotermes flavicollis Fabricius (1793) and Associated Aspergillus Micheli ex Haller (1768) Species.

The yellow-necked dry-wood termite Kalotermes flavicollis (Fabricius, 1793) (Blattodea: Kalotermitidae) is an important pest that infests wood in Europe. An increase in attacks by K. flavicollis has been seen in buildings and in churches in Palermo (Italy), with attacks on both structural elements and artefacts. Future climate changes are expected to lead to increasing temperatures, which will probably affect the pest status of this species, which is difficult to control. In this context, it is important to identify potential natural antagonists of K. flavicollis. During a survey of the K. flavicollis population, several dead individuals with evident fungal efflorescence were found. Therefore, a study aimed at the isolation and identification of these microorganisms was conducted. Fungal colonies isolated from mycelial structures grown on insects were identified based on morphological characteristics and DNA profiling. Three different species were identified: Aspergillus nomius, A. subramanianii, and A. tamarii. This is the first time that fungi have been recorded in association with the yellow-necked dry-wood termite and, in addition, this study reports the first association of A. subramanianii with Isoptera.

Oxidation Products of Tryptophan and Proline in Adipokinetic Hormones-Artifacts or Post-Translational Modifications?

Adipokinetic hormones (AKHs) regulate important physiological processes in insects. AKHs are short peptides with blocked termini and Trp in position 8. Often, proline occupies position 6. Few post-translational modifications have been found, including hydroxyproline ([Hyp6]) and kynurenine. Our recent data suggest that the Hyp- and Kyn-containing AKHs occur more often than originally thought and we here investigate if they are natural or artifactual. From crude extracts of the corpora cardiaca (CC) of various insect species, AKHs were analyzed using liquid chromatography coupled to high-resolution mass spectrometry (LC-MS). Synthetic [Hyp6]-AKHs were tested in an in vivo metabolic assay. Freshly dissected Periplaneta americana and Blaberus atropos CCs (with precautions taken against oxidation) were analyzed. B. atropos CC were placed into a depolarizing saline and the released AKHs were measured. Hyp was detected in several decapeptides from cockroaches. The modified form accompanied the AKH at concentrations below 7%. The [Hyp6]-AKHs of B. atropos were present in fresh CC preparations and were shown to be releasable from the CC ex vivo. Synthetic [Hyp6]-containing peptides tested positively in a hypertrehalosemic bioassay. Hydroxyprolination was also detected for Manto-CC from the termite Kalotermes flavicollis and for Tetsu-AKH of the grasshopper, Tetrix subulata. Oxidized Trp-containing forms of Nicve-AKH were found in species of the burying beetle genus Nicrophorus. Trp oxidation is known to occur easily during sample handling and is likely the reason for the present findings. For hydroxyprolination, however, the experimental evidence suggests endogenous processes.

Use of the proteomic tool MALDI-TOF MS in termite identification

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) has proved effective for the identification of many arthropods. A total of 432 termite specimens were collected in Mali, Cote d’Ivoire, Togo, Senegal, Switzerland and France. Morphologically, 22 species were identified, including Ancistrotermes cavithorax, Amitermes evuncifer, Cryptotermes brevis, Cubitermes orthognathus, Kalotermes flavicollis, Macrotermes bellicosus, Macrotermes herus, Macrotermes ivorensis, Macrotermes subhyalinus, Microcerotermes parvus, Microtermes sp., Odontotermes latericius, Procubitermes sjostedti, Promirotermes holmgreni, Reticulitermes grassei, Reticulitermes lucifugus, Reticulitermes santonensis, Trinervitermes geminatus, Trinervitermes occidentalis, Trinervitermes togoensis, Trinervitermes sp., Trinervitermes trinervoides and Trinervitermes trinervius. Analysis of MALDI-TOF MS spectra profiles from termites revealed that all were of high quality, with intra-species reproducibility and inter-species specificity. Blind testing of the spectra of 389 termites against our updated database with the spectra of 43 specimens of different termite species revealed that all were correctly identified with log score values (LSVs) ranging from 1.65 to 2.851, mean 2.290 ± 0.225, median 2.299, and 98.4% (383) had LSVs > 1.8. This study is the first on the use of MALDI-TOF for termite identification and shows its importance as a tool for arthropod taxonomy and reinforces the idea that MALDI-TOF MS is a promising tool in the field of entomology.

Identification of Cryptotermes brevis (Walker, 1853) and Kalotermes flavicollis (Fabricius, 1793) Termite Species by Detritus Analysis

We carried out morphological and dimensional analysis of the detritic elements deposited in the galleries of two termite species of the Kalotermitidae family present in Spain known as drywood termites (Cryptotermes brevis (Walker, 1853) and Kalotermes flavicollis (Fabricius, 1793)). This was in order to gauge the possibility of differentiating the species only on the basis of debris observation and analysis. Ten samples from six different geographical sources were analyzed and measured. Significant statistical differences were found between these two termite species in all measured parameters, and multivariate statistical models, able to predict species on the basis of dimensional measurements, were developed, with a degree of success higher than 75%. The most important dimensional differences were length and width, as well as the variable hexagonal shape of the cross-section of the detritic elements. The detritic elements of both species had a variable form of a hexagonal prism with slightly concave faces, and with pointed or rounded ends. Those of the Cryptotermes brevis species were significantly larger, less elongated, and had a smaller concavity on the faces of the prism. Color was found to be particularly variable in both species, and was not useful. Nevertheless, the debris of Kalotermes flavicollis was markedly “dirty” (mixed with other wood remains).

Molecular systematics, biogeography, and colony fusion in the European dry-wood termites Kalotermes spp. (Blattodea, Termitoidae, Kalotermitidae).

European dry-wood termites belong to the genus Kalotermes (Kalotermitidae), one of the two termite genera in Europe. Until the recent description of two new species, Kalotermes italicus in Italy and Kalotermes phoenicae in the eastern Mediterranean area, Kalotermes flavicollis was the only taxon known in this region. The presence of additional entities, suggested by morphological and physiological variation observed in K. flavicollis, was supported by molecular studies revealing four distinct genetic lineages: lineage A, K. flavicollis sensu strictu, from the Aegean area to Italy; lineage B, in Tuscany; lineage SC, in Sardinia and Corsica; lineage SF, in southern France. Lineages A and B may form mixed colonies, suggesting hybridization. To draw a more detailed picture of Kalotermes evolution and biogeography in Europe, we analyzed samples from previously unsampled areas, such as Spain and southern Italy, by means of the highly informative cox1/trnL/cox2 mitochondrial DNA marker. Overall, phylogenetic analyses confirmed previously identified lineages and taxa, but widened the distribution of the lineage SC to the mainland and of the lineage SF to Spain and Portugal. Results further provided evidence for the synonymy between lineage B and K. italicus. Species delimitation analysis suggested that the three K. flavicollis lineages, as well as K. italicus, can be separate taxa. Data also suggest a possible interspecific hybridization between K. italicus and both K. flavicollis lineages A and SC.

List of trees and shrubs infested by Kalotermes flavicollis (Kalotermitidae) in Greece

In Greece, the damage caused to trees and shrubs by termites has been increasing over the last 10 years. Local government and citizens have become interested in this pest due to frequent accidents caused by trees falling onto pavements, cars and people. This led the authors to carry out a survey to list the termite‐infested plant species, mainly on urban streets and in fields all over the country. The only termite species found was Kalotermes flavicollis (family Kalotermitidae). The present study highlights the current situation in Greece and provides a list of the fruit and ornamental trees and shrubs found to be infested by K. flavicollis, during the survey.

Polyamidoamines (PAAs) functionalized with siloxanes as wood preservatives against fungi and insects

Abstract A novel treatment based on polyamidoamines (PAAs) for the preservation of wood against fungi and insects with a broad protection functionality, low effective concentration, and low environmental impact has been developed. PAAs were synthesized by nucleophile addition of ethanolamine (EtA) and/or 3-aminopropyltriethoxysilane (APTES) to N,N′-methylene-bisacrylamide (MBA). The molar ratios in the tested formulation were: I) MBA:EtA=1:1; II) MBA:APTES=1:1; III) MBA:EtA:APTES=1:0.5:0.5. These formulations, characterized by ESI-MS, NMR, FT-IR, were tested against: (a) the wood decay fungi (Coniophora puteana, Coriolus versicolor, and Poria placenta); (b) the subterranean termite Reticulitermes lucifugus, and the drywood termite Kalotermes flavicollis; (c) the woodborer Stegobium paniceum. Tests in combination with leaching showed that formulation II and III can be utilized as fungal wood preservatives for use classes 2–3 (EN 335:2013). In addition, all PAAs formulations were equally effective in preserving wood against the subterranean termite, and formulation II was most effective against drywood termite. On the other hand, the formulation I showed good efficacy against S. paniceum.

Molecular characterization and phylogeny of Kalotermes populations from the Levant, and description of Kalotermes phoeniciae sp. nov.

The presence of the yellow-necked drywood termite, Kalotermes flavicollis Fabr., has been reported along most of the Mediterranean coasts of Europe, Africa and Asia. While morphological and genetic data exist on European K. flavicollis populations from western and central Mediterranean regions, data on eastern Mediterranean Kalotermes are scarce, and no genetic data exist to date. In this study, we analyzed 17 Kalotermes sp. colonies from 11 localities in the Levant (Cyprus, Lebanon, and Israel), in order to characterize genetically (mitochondrial DNA: COII, 16S, and control region) these populations. We found that samples from the Levant are genetically different from K. flavicollis, with distance values falling in the range of interspecific distances. In the phylogeny of European Kalotermes populations, samples from the Levant form a clade of their own, sister to a clade including K. flavicollis and Kalotermes italicus. Inside the eastern Mediterranean clade, all the samples from Cyprus are included in a well-supported subclade, suggesting that the colonization of the island might have occurred in a single event. These findings show that the populations we examined do not belong to the species K. flavicollis, but to a new species peculiar to the Levant, that we describe as Kalotermes phoeniciae sp. nov. It is possible that previous reports of K. flavicollis in this region can be attributed to K. phoeniciae.

Phylogeny and Ultrastructure of Oxymonas jouteli, a Rostellum-free Species, and Opisthomitus longiflagellatus sp. nov., Oxymonadid Flagellates from the Gut of Neotermes jouteli

The biodiversity of oxymonadid flagellates in termite hindguts is not fully explored. Many species have been differentiated only by morphological features, and small-celled species have been overlooked or ignored. Our analysis of the dry wood termite Neotermes jouteli by light and electron microscopy revealed the presence of two distinct morphotypes of oxymonads. The larger one matched the morphology of Oxymonas jouteli, the only oxymonad species described from this termite. Although it generally lacks the typical anterior rostellum of the genus Oxymonas, its SSU rRNA gene sequence clusters among other members of this genus, including novel phylotypes that we obtained from Incisitermes tabogae. The second morphotype was a tiny oxymonad that showed the typical traits of the genus Opisthomitus, including a pointed anterior prolongation (lappet). However, the four equal flagella were much longer than those of Opisthomitus avicularis from Kalotermes flavicollis, the only species of the genus and so far described only by light microscopy. We provide a detailed description of Opisthomitus longiflagellatus sp. nov. and demonstrate that despite ultrastructural similarities to members of the Polymastigidae, its SSU rRNA gene sequences form a separate family-level lineage with a slight affinity to the Pyrsonymphidae.

Microwaves as a Remedial Treatment of Wood

A pilot experiment of microwave treatment was carried out on an Art Nouveau residential building erected at the beginning of the 20th century in Valencia, Spain. The timber structure was affected by a combined damage caused by Anobium punctatum (common furniture beetle or common house borer) and Kalotermes flavicollis (yellownecked dry-wood termite). After performing an exhaustive preliminary study on the building and analyzing the different alternatives of wood treatment, it was decided to use a microwave machine especially designed to eliminate this type of wood boring insects.This machine generates microwaves, a type of electromagnetic radiation, with a frequency of 2.45 GHz. This circumstance makes it possible to produce a high-frequency alternating electric field that causes the rotation of the molecular dipoles of water. Because of this continuous rotation the temperature of the water rises considerably.By heating up the humidity inside a living being, as in the case of xylophagous insects, an artificial fever in the body is generated, and since the animal is not capable of tolerating this temperature, it dies.In order to eliminate xylophagous insects, the wood moisture content (usually between 10% and 12%) and the presence of water in wood boring insects (around 90%) must be taken into account. Due to the greater content of humidity the increase of the temperature is higher and quicker in insects than wood.After having studied and experimented with different combination of power and time of exposition to the microwaves, this machine was found capable of eliminating any type of insect that may live inside the timber without harming the wood itself.Prior to this pilot intervention, several research works were carried out with the same microwave machine, both on structural timber and wooden works of art. In all these studies it was observed that no damage was caused either to the wood or to the finish of the works of art including pigments, polishes, stains etc. On the other hand, 100% efficacy was achieved in the elimination of xylophagous insects.This article describes the design, experimentation and perfecting process of this microwave machine and its pilot application on structural timber in an Art Nouveau residential building. As an example of method, the intervention followed the main steps of diagnosis and treatment testing including the analysis of several options of intensity and time that led to the positive final results obtained.

THE COMPETENCE OF KALOTERMES FLAVICOLLIS FABR. (ISOPTERA) PSEUDERGATES TO DIFFERENTIATE INTO SOLDIERS

SUMMARY Previous investigations carried out in our laboratory on Kalotermes flavicollis Fabr. had shown that the royal pair exerts a stimulatory action on pseudergate differentiation into soldiers, while the latter have an inhibitory influence on it (SPRINGHETTI, 1969). The present experiments have shown that the pseudergate moulting interval is of fairly variable duration, from 18 to 140 days, with a mean of about 70 days. Its average duration, however, is the same both in specimens differentiating into soldiers and in those undergoing a larval moult. Pseudergates display a rather short competence period for differentiation into soldiers during the second half of the moulting interval: possibly between the 45th and 60th day. Pseudergates are responsive only during their competence period, to the antagonistic stimuli exerted by the royals and soldiers respectively. The absence of soldiers prior to the 40th day does not elicit the differentiation of new soldiers (even in the presence of the reproductives);...

Extreme genetic mixing within colonies of the wood‐dwelling termiteKalotermes flavicollis(Isoptera,Kalotermitidae)

The existence of altruism in social insects is commonly attributed to altruistic individuals gaining indirect fitness through kin selection. However, recent studies suggest that such individuals might also gain direct fitness through reproduction. Experimental studies on primitive wood-dwelling termites revealed that colony fusion often causes the death of primary reproductives (queen and king), allowing opportunities for workers to inherit the nest by developing into replacement reproductives (neotenics). Therefore, colony fusion has been proposed as an important factor that may have favoured sociality in termites. However, whether colony fusion occurs frequently in natural populations of wood-dwelling termites remains an open question. We analysed eleven colonies of the wood-dwelling termite Kalotermes flavicollis (Kalotermitidae), using two mitochondrial and five nuclear microsatellite markers. Nine of eleven colonies (82%) were mixed families, with offspring of three or more primary reproductives. To our knowledge, this result represents the highest frequency of mixed-family colonies ever reported in termites. Moreover, genetic mixing of colonies appeared extreme in two ways. First, the number of haplotypes per colony was exceptionally high (up to nine), indicating that colonies were composed of multiple queens' offspring. Second, some mixed-family colonies included individuals belonging to two highly divergent genetic lineages. F-statistics and relatedness values suggest that mixed-family colonies most likely result from colony fusion, giving support to the accelerated nest inheritance theory. These findings raise important questions about the mode of foundation of mixed-family colonies and the evolutionary forces that maintain them within populations.

A Dark-Necked Drywood Termite (Isoptera: Kalotermitidae) in Italy: Description ofKalotermes italicussp. nov.

The yellow-necked drywood termite, Kalotermes flavicollis (F.), so called due to the yellow pronotum of its alates, is the only species in the genus Kalotermes known for Europe. In some Italian localities, K. flavicollis swarms can contain a small proportion of alates with a dark pronotum, but otherwise not different from the normally colored K. flavicollis. This color variation was described by G. Becker in 1955 as Kalotermes flavicollis var. fuscicollis. During collecting trips in central Italy, we found in Grosseto Marina (Tuscany) a Kalotermes population whose alates all have a very dark pronotum. Compared with K. flavicollis alates, Grosseto alates also have paler wings and smaller arolia. Grosseto soldiers have eyes smaller than those of K. flavicollis soldiers. Kalotermes sp. form Grosseto also differs morphologically from the other Kalotermes species known for the circum-Mediterranean lands. The population from Grosseto has mitochondrial DNA sequences (a partial sequence of the control region and a fragment including a portion of COI, tRNA-Leu and a portion of COII) quite different from K flavicollis (p-distance: 5.6–7.3%). Some Kalotermes populations from Tuscany and Marche, morphologically classifiable as K flavicollis, have sequences similar to those of Grosseto Kalotermes (p-distance: 0.1–1.0%). These populations are possibly hybrids between K. flavicollis and Kalotermes sp. from Grosseto. Because of its morphologic and genetic distinctive features, we describe Kalotermes sp. from Grosseto as Kalotermes italicus sp. nov.

Evaluation of polyphenols-rich natural compounds as treatments to prevent attacks by subterranean and drywood termites: preliminary results

In the view to find sustainable methods to prevent termite attacks to wooden objects and structural timbers, this study represents a preliminary step in the evaluation of some natural substances considered as effective by some African popular traditions. Dark shea cake, obtained from the kernels of <em>Vitellaria paradoxa </em>(Sapotaceae), is the phase just before the extraction of shea (= karitè) butter. In some West African regions, by-products from this extraction are traditionally believed to protect houses from termites. To verify if this practice has scientific basis, shea cake was used in experiments with both subterranean and drywood termites, respectively <em>Reticulitermes lucifugus </em>(Rossi) (Rhinotermitidae) and <em>Kalotermes flavicollis </em>(Fabricius) (Kalotermitidae). As shea nuts are extremely rich in polyphenols, the trials included a comparison with tannins from chestnut (<em>Castanea sativa</em>). Short-term experiments to evaluate repellency and<em> </em>feeding deterrence of the two compounds were performed. Results showed differences in the behavior of the two termites species and that shea cake is significantly more repellent than chestnut tannins, especially for <em>K. flavicollis</em>. No feeding deterrence activity was detected for either substance for either termite species. Further investigation is currently being performed to better clarify how these compounds affect termite biology.

Wood impregnated with metal chelates dissolved in organic media tested for termite resistance

Wood manufactured products are subjected to biological decay due to fungi and insects. The use of copper chelates as biocides was proposed, due to their high stability which minimizes copper leaching into the environment. Considering the remarkable effectiveness showed by copper chelates on brown rot fungi, zinc and copper salicylate complexes were prepared in order to have metal chelates soluble in organic media available. The present study aimed at evaluating these metal chelates complexes as preservative agents for wood treatment against termites. Trials were performed on <em>Reticulitermes lucifugus</em> (Rossi) and <em>Kalotermes flavicollis</em> (Fabricius). Results showed that in both termite species wood consumption was significantly lower on Cu-chelates treated samples compared to untreated wood, whereas the wood slices impregnated with Zn-chelates and the organic media alone gave an intermediate response. Interestingly, in one case solvent-impregnated wood was significantly more attractive than untreated wood for both species and further investigations are being carried out to clarify this behaviour.

The Non-LTR Retrotransposon R2 in Termites (Insecta, Isoptera): Characterization and Dynamics

The full-length element of the non-LTR retrotransposon R2 is here characterized in three European isopteran species: the more primitive Kalotermes flavicollis (Kalotermitidae), including two highly divergent mitochondrial lineages, and the more derived Reticulitermes lucifugus and R. urbis (Rhinotermitidae). Partial 3' sequences for R. grassei and R. balkanensis were also analyzed. The essential structural features of R2 elements are conserved in termites. Phylogenetic analysis revealed that termite elements belong to the same clade and that their phylogeny is fully compatible with the phylogeny of their host species. The study of the number and the frequency of R2 insertion variants in four R. urbis colonies suggests a greatly reduced, or completely absent, recent element activity.

Mitochondrial and nuclear markers highlight the biodiversity of Kalotermes flavicollis (Fabricius, 1793) (Insecta, Isoptera, Kalotermitidae) in the Mediterranean area

The biodiversity of the European termite Kalotermes flavicollis is here studied through the analysis of mitochondrial (303 bp of control region and 912 bp of COI/tRNA(Leu)/COII) and nuclear (five microsatellite and 20 Inter-SINE loci) markers on 18 colonies collected in Southern France, Corsica, Sardinia, peninsular Italy, the Balkans and Greece. Different statistical analyses (Bayesian phylogenetic analysis,parsimony network, F-statistics, PCA) were performed. Mitochondrial sequences produced an unresolved polytomy including samples from peninsular Italy, Balkans and Greece, and three main clades: southern France, Corsica-Sardinia and Portoscuso(SW Sardinia). Nuclear markers confirm these data, further highlighting a more significant divergence at the regional scale. The results obtained for the peri-Tyrrhenian area agree with major paleogeographic and paleoclimatic events that shaped the biodiversity of the local fauna. K. flavicollis biodiversity and its phylogeographic pattern are also evaluated in the light of the data available for the other native European termite taxon (genus Reticulitermes), in order to produce a more complete scenario of the Mediterranean. In the area comprised between southern France and Italy, the degree of diversity is similar; however, in the eastern area, while K. flavicollis is differentiated only at the population level, the genus Reticulitermes comprises at least six entities of specific and/or subspecific level. This discrepancy may be explained by taking into account the different evolutionary histories of the two taxa.

Identification and localization of the multiple bacterial symbionts of the termite gut flagellate Joenia annectens

The hindgut of wood-feeding lower termites is densely colonized by a multitude of symbiotic micro-organisms. While it is well established that the eukaryotic flagellates play a major role in the degradation of lignocellulose, much less is known about the identity and function of the prokaryotic symbionts associated with the flagellates. Our ultrastructural investigations of the gut flagellate Joenia annectens (from the termite Kalotermes flavicollis) revealed a dense colonization of this flagellate by diverse ecto- and endosymbiotic bacteria. Phylogenetic analysis of the small-subunit rRNA gene sequences combined with fluorescence in situ hybridization allowed us to identify and localize the different morphotypes. Furthermore, we could show that K. flavicollis harbours two phylotypes of J. annectens that could be distinguished not only by their small-subunit rRNA gene sequences, but also by differences in their assemblages of bacterial symbionts. Each of the flagellate populations hosted phylogenetically distinct ectosymbionts from the phylum Bacteroidetes, one of them closely related to the ectosymbionts of other termite gut flagellates. A single phylotype of 'Endomicrobia' was consistently associated with only one of the host phylotypes, although not all individuals were colonized, corroborating that 'Endomicrobia' symbionts do not always cospeciate with their host lineages. Flagellates from both populations were loosely associated with a single phylotype of Spirochaetales attached to their cell surface in varying abundance. Current evidence for the involvement of Bacteroidales and 'Endomicrobia' symbionts in the nitrogen metabolism of the host flagellate is discussed.

Seasonal Trends in the Social Composition and Inside-Trunk Distribution ofKalotermes flavicollis(Isoptera: Kalotermitidae) Colonizing Grapevines

The polyphagous drywood termite Kalotermes flavicollis (Fabricius) can establish its colonies inside living trees, causing a progressive debilitation that ends with the death of the plant. For this reason, it is considered one of the major pests of Sherry vineyards in Andalusia, SW Spain. To analyze the trends of colonization of this termite, a detailed investigation was performed during 16 mo in seven districts of the Sherry area, cutting randomly selected grapevine trunks into six sections, from which all individuals were collected and counted according to their developmental instar or caste. The results detected variations in the social composition of the groups colonizing grapevines according to the spatial distribution inside the vine trunk and in the numerical trends over the seasons. A pattern of seasonal migrations of the termites inside the trunk was inferred, with upward movements from spring to autumn and downward movements from autumn to winter. During winter, when termites were mainly located in the medium-basal sections of the trunk, a higher proportion of eggs and larvae was detected. Going from spring to summer, when the majority of the termites was located in the medium-upper sections of the trunk, there was an increase of nymphal instars followed, during autumn, by the appearance of alates, mostly located in the upper sections of the grapevine. Pseudergates and reproductives maintained their proportions and locations almost unchanged during the year.

Iridoid Glucosides with Insecticidal Activity from Galium melanantherum

The insecticidal activity of the endemic species Galium melanantherum was evaluated against Crematogaster scutellaris ants and Kalotermes flavicollis termites. Iridoid glucosides 1-7 were isolated for the first time as metabolites of the investigated plant, along with the coumarin scopolin. The main components of the extract were found to be the non-acetylated iridoids: geniposidic acid (1), 10-hydroxyloganin (2), deacetyldaphylloside (3), monotropein (4), deacetylasperulosidic acid (5) and scandoside (6), while asperulosidic acid (7) was present only in minute quantities. All isolated metabolites were identified on the basis of their spectral data. Laboratory bioassays revealed significant levels of toxicity for 1-4 against Kalotermes flavicollis termites and Crematogaster scutellaris ants.