Chemical Communication Systems Research Articles

Simple and rapid CE‐UV method for the assessment of trail pheromone compounds of leaf‐cutting ants' venom glands

Leaf-cutting ants cause large losses in several crops around the world. In the ant species Atta sexdens, each colony comprises up to 5 million individuals. In order to keep a close connection among such a large number of individuals, an efficient chemical communication system is necessary. Among other different substances, these animals use alkylpyrazines to mark their trails and to guide ant workers from the nest to their sources of food. In this study, CE-UV was used to apply a method for qualitative analysis of venom gland components of leaf-cutting ants. Mobility of these compounds proved to be a function of the ionization capability of these bases as well as their volumes. Migration order was thoroughly explained in terms of such parameters. The best analysis conditions were achieved with a BGE composed by 0.8% formic acid plus 20% methanol in water, hydrodynamic injection, and application of external pressure. Such analysis conditions may be easily applied in CE-MS analyses as well. CE-UV analyses proved to be as adequate as GC to analyze such compounds due to system detectability (LD ≅ 0.005 mmol/L), separation efficiency (from 5.07×10(4) to 1.23×10(5) theoretical plates), and resolution (minimum of 2.35). In addition, analysis time was ca. 15 min, which shows another advantage of CE analysis when compared with GC. Although the analytes are found in concentrations as low as 50 ng/venom glands, four putative pyrazine ring moiety substances could be detected in real samples, due to sample stacking and use of a capillary with extended detection cell.

Venom volatiles of the paper wasp social parasite Polistes sulcifer elicit intra-colonial aggression on the nest of the host species Polistes dominulus

Insect social parasites rely on host workers to rear and protect their own brood. To conquer a host colony, a parasite must overcome the defensive mechanisms of the host, often by exploiting its chemical communication system. A widespread strategy involves the production of specific allomones (the so-called “propaganda pheromones”) to facilitate the usurpation process by manipulating the defensive behavior of the host. Polistes sulcifer is the obligate and permanent social parasite of the congeneric paper wasp Polistes dominulus. In this study, we investigated if the venom volatiles, well known to be alarm pheromones in the host species, could be used by the parasite to manipulate the host defense. We thus performed laboratory bioassays, to evaluate the possible effect of the venom volatile compounds of the parasite on the host. Our results show that host colony members reacted to the venom volatiles extract of the parasite with an increase in intra-colonial aggression compared to the reaction induced by the venom volatiles extract of the host foundress. Besides, a re-analysis of previously published chemical data showed that the parasite venom volatiles profile differs from that of the host: the spiroacetals are absent, whilst the amides are very abundant in the parasite venom when compared with that of the host. Similar to other insect social parasites, Polistes wasp parasites might be able to increase their invasion success by using venom volatile pheromones to distract the host defenders.

Task Group Differences in Cuticular Lipids in the Honey Bee Apis mellifera

Social insects are defined by their ability to divide labor among their numerous nestmates. This is achieved via a complex system of chemical communication that allows colonies to organize task activity so as to maximize the productivity of the colony. However, the mechanism by which social insects distinguish task groups among morphologically identical individuals remains unknown. Using the honey bee, Apis mellifera, as our model species, we investigated the presence of task-specific patterns in the cuticular lipids (n-alkanes, fatty acids, and alkenes) of bees. Cuticular lipids are known to play an essential role in the recognition processes of insects. We found task-specific features in the n-alkane and alkene profiles of bees, but no task-specific patterns in the fatty acid profile. Foragers, in particular, had elevated levels of n-alkanes relative to nurse and newly emerged bees, suggesting increased waterproofing. Newly emerged bees had low levels of cuticular lipids, supporting the Blank Slate theory and potentially explaining their acceptance into foreign colonies.

Chemical Profiles of Body Surfaces and Nests from Six Bornean Stingless Bee Species

Stingless bees (Apidae: Meliponini) are the most diverse group of Apid bees and represent common pollinators in tropical ecosystems. Like honeybees they live in large eusocial colonies and rely on complex chemical recognition and communication systems. In contrast to honeybees, their ecology and especially their chemical ecology have received only little attention, particularly in the Old World. We previously have analyzed the chemical profiles of six paleotropical stingless bee species from Borneo and revealed the presence of species-specific cuticular terpenes- an environmentally derived compound class so far unique among social insects. Here, we compared the bees' surface profiles to the chemistry of their nest material. Terpenes, alkanes, and alkenes were the dominant compound groups on both body surfaces and nest material. However, bee profiles and nests strongly differed in their chemical composition. Body surfaces thus did not merely mirror nests, rendering a passive compound transfer from nests to bees unlikely. The difference between nests and bees was particularly pronounced when all resin-derived compounds (terpenes) were excluded and only genetically determined compounds were considered. When terpenes were included, bee profiles and nest material still differed, because whole groups of terpenes (e.g., sesquiterpenes) were found in nest material of some species, but missing in their chemical profile, indicating that bees are able to influence the terpene composition both in their nests and on their surfaces.

Queen pheromones

Group-living species produce signals that alter the behavior and even the physiology of their social partners. Social insects possess especially sophisticated chemical communication systems that govern every aspect of colony life, including the defining feature of eusociality: reproductive division of labor. Current evidence hints at the central importance of queen pheromones, but progress has been hindered by the fact that such pheromones have only been isolated in honeybees. In a pair of papers on the ant Lasius niger, we identified and investigated a queen pheromone regulating worker sterility. The cuticular hydrocarbon 3-methylhentriacontane (3-MeC31) is correlated with queen maturity and fecundity, and workers are also more likely to execute surplus queens that have low amounts of this chemical. Experiments with synthetic 3-MeC31 found that it inhibits ovarian development in queenless workers and lowers worker aggression towards objects coated with it. Production of 3-MeC31 by queens was depressed by an experimental immune challenge, and the same chemical was abundant on queen-laid eggs, suggesting that the workers’ responses to the queen are conditional on her health and fecundity. Together with other studies, these results indicate that queen pheromones are honest signals of quality that simultaneously regulate multiple social behaviors.

Ontogeny of Drosophila melanogaster female sex‐appeal and cuticular hydrocarbons

The chemical communication system in Drosophila melanogaster Meigen, 1830 plays a major role in courtship and consists of the male-specific cis-Vaccenyl acetate and sex-specific contact pheromones, cuticular hydrocarbons (CHC), which build up during ontogeny (first 4 days). They replace longer CHCs, common to both sexes and present only after the imaginal eclosion. A detailed quantitative description of the evolution of cuticular unsaturated hydrocarbons with age is presented here for males and females of different D. melanogaster strains, which have been bred in well controlled environments. Monoenes appear in both sexes at around 12 h, before female dienes. The present paper argues that this is likely linked to the switching on of a new set of genes. Ecdysone, which is more abundant in females than in males during this critical period, might control this switch. Parallel behavioral studies show that whereas female of all ages trigger early mature male courtship steps like wing vibration, only females older than 1 day trigger late courtship steps like attempted copulation. This supports the hypothesis that late male courtship steps might be triggered by the CHCs, which build up after this age, especially female-specific (Z,Z)-7,11 - and (Z,Z)-5,9-dienes.

Plant volatiles

Plant volatiles are the metabolites that plants release into the air. The quantities released are not trivial. Almost one-fifth of the atmospheric CO2 fixed by land plants is released back into the air each day as volatiles. Plants are champion synthetic chemists; they take advantage of their anabolic prowess to produce volatiles, which they use to protect themselves against biotic and abiotic stresses and to provide information - and potentially disinformation - to mutualists and competitors alike. As transferors of information, volatiles have provided plants with solutions to the challenges associated with being rooted in the ground and immobile.

Behavioral responses of Steirastoma breve (Sulzer) (Coleoptera: Cerambycidae) to host plant Theobroma cacao L., brushwood piles, under field conditions

Steirastoma breve is one of the most important pests in cocoa plantations in the Neotropics. Preliminary studies suggest that the chemical communication system in S. breve may be modulated by kairomones produced by cocoa plants and pheromones released by males. We examined the role of Theobroma cacao brushwood in the communication system of S. breve under field conditions. Sixty kilograms of T. cacao and Bauhinia sp. branches were cut, grouped into separate piles, and placed in an experimental cocoa plantation as an odor source. The sex and number of S. breve adults landing on each pile were registered during 23 days from 9:00 to 15:00 h. Each individual arriving at a pile was marked with a coded color and then released onto the same pile. All behavioral activities undertaken by S. breve were evaluated and flight behavior towards the T. cacao piles was described. S. breve adults were attracted to T. cacao piles placed under both sunny and shaded conditions, but were not attracted to Bauhinia sp. S. breve males dispersed more in shaded piles than in those placed in the sun, and the proportion of encounters between sexes was higher in the piles at which they had first arrived. Analysis of the data confirmed the presence and importance of T. cacao kairomones in the chemical communication system of this important cocoa pest.

Characterization of an Enantioselective Odorant Receptor in the Yellow Fever Mosquito Aedes aegypti

Enantiomers differ only in the left or right handedness (chirality) of their orientations and exhibit identical chemical and physical properties. In chemical communication systems, enantiomers can be differentially active at the physiological and behavioral levels. Only recently were enantioselective odorant receptors demonstrated in mammals while their existence in insects has remained hypothetical. Using the two-microelectrode voltage clamp of Xenopus oocytes, we show that the yellow fever mosquito, Aedes aegypti, odorant receptor 8 (AaOR8) acts as a chiral selective receptor for the (R)-(—)-enantiomer of 1-octen-3-ol, which in the presence of other kairomones is an attractant used by blood-sucking insects to locate their hosts. In addition to steric constraints, chain length and degree of unsaturation play important roles in this recognition process. This is the first characterization of an enantioselective odorant receptor in insects and the results demonstrate that an OR alone, without helper proteins, can account for chiral specificity exhibited by olfactory sensory neurons (OSNs).

Sex-specific antennal sensory system in the ant Camponotus japonicus: structure and distribution of sensilla on the flagellum

The antennae are a critically important component of the ant's highly elaborated chemical communication systems. However, our understanding of the organization of the sensory systems on the antennae of ants, from peripheral receptors to central and output systems, is poorly understood. Consequently, we have used scanning electron and confocal laser microscopy to create virtually complete maps of the structure, numbers of sensory neurons, and distribution patterns of all types of external sensilla on the antennal flagellum of all types of colony members of the carpenter ant Camponotus japonicus. Based on the outer cuticular structures, the sensilla have been classified into seven types: coelocapitular, coeloconic, ampullaceal, basiconic, trichoid-I, trichoid-II, and chaetic sensilla. Retrograde staining of antennal nerves has enabled us to count the number of sensory neurons housed in the different types of sensilla: three in a coelocapitular sensillum, three in a coeloconic sensillum, one in an ampullaceal sensillum, over 130 in a basiconic sensillum, 50-60 in a trichoid-I sensillum, and 8-9 in a trichoid-II sensillum. The basiconic sensilla, which are cuticular hydrocarbon-receptive in the ant, are present in workers and unmated queens but absent in males. Coelocapitular sensilla (putatively hygro- and thermoreceptive) have been newly identified in this study. Coelocapitular, coeloconic, and ampullaceal sensilla form clusters and show biased distributions on flagellar segments of antennae in all colony members. The total numbers of sensilla per flagellum are about 9000 in unmated queens, 7500 in workers, and 6000 in males. This is the first report presenting comprehensive sensillar maps of antennae in ants.

Microbial Communication, Cooperation and Cheating: Quorum Sensing Drives the Evolution of Cooperation in Bacteria

An increasing body of empirical evidence suggests that cooperation among clone-mates is common in bacteria. Bacterial cooperation may take the form of the excretion of “public goods”: exoproducts such as virulence factors, exoenzymes or components of the matrix in biofilms, to yield significant benefit for individuals joining in the common effort of producing them. Supposedly in order to spare unnecessary costs when the population is too sparse to supply the sufficient exoproduct level, many bacteria have evolved a simple chemical communication system called quorum sensing (QS), to “measure” the population density of clone-mates in their close neighborhood. Cooperation genes are expressed only above a threshold rate of QS signal molecule re-capture, i.e., above the local quorum of cooperators. The cooperative population is exposed to exploitation by cheaters, i.e., mutants who contribute less or nil to the effort but fully enjoy the benefits of cooperation. The communication system is also vulnerable to a different type of cheaters (“Liars”) who may produce the QS signal but not the exoproduct, thus ruining the reliability of the signal. Since there is no reason to assume that such cheaters cannot evolve and invade the populations of honestly signaling cooperators, the empirical fact of the existence of both bacterial cooperation and the associated QS communication system seems puzzling. Using a stochastic cellular automaton approach and allowing mutations in an initially non-cooperating, non-communicating strain we show that both cooperation and the associated communication system can evolve, spread and remain persistent. The QS genes help cooperative behavior to invade the population, and vice versa; cooperation and communication might have evolved synergistically in bacteria. Moreover, in good agreement with the empirical data recently available, this synergism opens up a remarkably rich repertoire of social interactions in which cheating and exploitation are commonplace.

The Antennal Lobes of Fungus-Growing Ants (Attini): Neuroanatomical Traits and Evolutionary Trends

Ants of the tribe Attini are characterized by their obligate cultivation of symbiotic fungi. In addition to the complex chemical communication system of ants in general, substrate selection and fungus cultivation pose high demands on the olfactory system of the Attini. Indeed, behavioral studies have shown a rich diversity of olfactory-guided behaviors and tremendous odor sensitivity has been demonstrated. To allow fine-tuned behavioral responses, adaptations within the olfactory system of the Attini are expected. We compared the number, volumes and position of the glomeruli (functional units) of the antennal lobe of 25 different species from all three major Attini groups (lower, higher and leaf-cutting Attini). The antennal lobes of all investigated Attini comprise a high number of glomeruli (>257). The highest number (630) was found in Apterostigma cf. mayri. This species is at a basal position within the Attini phylogeny, and we suggest that a high number of glomeruli might have been advantageous in the evolution of the advanced olfactory systems of the Attini. In the leaf-cutting Attini, an extremely large glomerulus (macroglomerulus) near the antennal nerve entrance was recently described in two species. Preliminary results show that this macroglomerulus is involved in processing of trail-pheromone information. In our comparative study, we find this macroglomerulus in all investigated leaf-cutting Attini, but in none of the lower and higher Attini species. It is found only in large workers, and for all investigated species it is located close to the entrance of the antennal nerve. Our results indicate that the presence of a macroglomerulus in large workers of leaf-cutting Attini is a derived over-expression of a trait in the polymorphic leaf-cutting species. It presumably represents an olfactory adaptation to elaborate foraging and mass recruitment systems, and adds to the complexity of division of labor and social organization known for this group.

Chemical communication by infochemicals

Homo sapiens depends very much on visual and acoustic stimuli to get along in every day life whereas the role of odours is considered as of secondary importance. This is very different for most environmental organisms who live in an “odour world” and depend strongly on chemical stimuli to learn about their biotic and abiotic environments. These chemical cues are called infochemicals. The chemical environment plays a prevalent role for the fine tuning of articles that increase the survival of a species, such as searching food or shelter, reproducing, optimizing the survival conditions for the offspring or avoiding potential predators. Especially in aquatic systems, the dominance of the chemical sense has clear advantages as it allows to perceive stimuli at night as well as in turbid water, and visual obstacles are no hindrance for perception. The complexity and sophistication of the chemical communication systems are difficult to imagine from the human perspective (Burks and Lodge 2002; Bronmark and Hansson 2000; Dodson et al. 1994; Klaschka 2008a, b). Natural infochemicals are usually odorants. Therefore, the knowledge of the basic steps of the reaction cascade of the olfactory perception is needed to understand the chemical communication by infochemicals. A generalized scheme is depicted in Fig. 1. Infochemicals are released by a biotic or abiotic sender. They may find the olfactory epithelium of a receiver where they can bind to specific odorant-binding proteins leading to the formation of an electrical signal which is transduced to the central nervous system evoking the perception. The perception of an infochemical can lead to morphological, physiological or behavioral reactions of the receiver or even to changes on population levels. Internal factors (such as age, sex, activity state) and external factors (such as time of the day or the year, temperature) can influence the emission of infochemicals by the sender as well as the various steps of the reaction cascade in the receiver.

GEOGRAPHIC AND TEMPORAL VARIATION IN MOTH CHEMICAL COMMUNICATION

In moth pheromone communication signals, both quantitative and qualitative intraspecific differences have been found across geographic regions. Such variation has generally been hypothesized to be due to selection, but evidence of genetic control of these differences is largely lacking. To explore the patterns of variation in pheromone signals, we quantified variation in the female sex pheromone blend and male responses of two closely related noctuid moth species in five different geographic regions for 2-3 consecutive years. We found significant variation in the ratios of sex pheromone blend components as well as in male response, not only between geographic regions but also within a region between consecutive years. The temporal variation was of a similar magnitude as the geographic variation. As far as we know, this is the first study reporting such temporal variation in moth chemical communication systems. The geographic variation seems to at least partly be controlled by genetic factors, and to be correlated with the quality of the local chemical environment. However, the pattern of temporal variation within populations suggests that optimization of the pheromonal signal also may be driven by within-generation physiological adjustments by the moths in response to their experience of the local chemical environment.

A new challenge—development of test systems for the infochemical effect

Many-if not all-organisms depend on so-called infochemicals, chemical substances in their surroundings which inform the receivers about their biotic and abiotic environment and which allow them to react adequately to these signals. Anthropogenic substances can interfere with this complex chemical communication system. This finding is called infochemical effect. So far, it is not known to what extent anthropogenic discharges act as infochemicals and influence life and reproduction of organisms in the environment because adequate testing methods to identify chemicals which show the infochemical effect and to quantify their effects have not been developed yet. The purpose of this article is to help and find suitable test designs. Test systems used in basic research to elucidate the olfactory cascade and the communication of environmental organisms by infochemicals are plentiful. Some of them might be the basis for a quantified ecotoxicological analysis of the infochemical effect. In principle, test systems for the infochemical effect could be developed at each step of the chemosensory signal transduction and processing cascade. Experimental set-ups were compiled systematically under the aspect whether they might be usable for testing the infochemical effect of single chemicals in standardized quantifying laboratory experiments. For an appropriate ecotoxicological assessment of the infochemical effect, experimental studies of many disciplines, such as molecular biology, neurobiology, physiology, chemical ecology, and population dynamics, should be evaluated in detail before a decision can be made which test system, respectively which test battery, might be suited best. The test systems presented here are based on the knowledge of the genetic sequences for olfactory receptors, binding studies of odorants, signal transmission, and reactions of the receivers on the level of the organisms or the populations. The following basic approaches are conceivable to identify the role of an infochemical: binding studies to the odorant-binding protein or to the odorant receptor binding protein (e.g., by in situ hybridization and immunohistochemical studies), measurement of electrical signals of the receptor cells in the tissue (e.g., electroolfactograms, electroantennograms), registration of phenotypic changes (e.g., observation under the microscope), behavioral tests (e.g., in situ online biomonitoring, use of T-shaped olfactometers, tests of avoidance responses), measurement of population changes (e.g., cell density or turbidity measurements), and multispecies tests with observation of community structure and community function. The main focus of this study is on aquatic organisms. It is evident that the infochemical effect is a very complex sublethal endpoint, and it needs further studies with standardized quantitative methods to elucidate whether and to what extent the ecosystem is affected. The collection of approaches presented here is far from being complete but should serve as a point of depart for further experimental research. This article is the first to compare various approaches for testing the infochemical effect. The development of a suitable test system will not be easy as there are a multitude of relevant chemicals, a multitude of relevant receptors, and a multitude of relevant reactions, and it must be expected that the effective concentrations are very low. The chemical communication is of utmost importance for the ecosystem and justifies great endeavors to find solutions to these technical problems. The infochemical effect is a new chapter in ecotoxicology. Will a new endpoint, the so-called infochemical effect, be required in addition to the actual standard test battery of Annex 5 to Commission Directive 92/69/EEC (EC 1992)? Finding the answer to this question is a big challenge that could be met by a comprehensive research project.

Population dynamics of Chaoborus flavicans and Daphnia spp.: effects on a zooplankton community in a volcanic eutrophic lake with naturally high metal concentrations (L. Monticchio Grande, Southern Italy)

The response of Daphnia populations to invertebrate predators involves morphological or behavioural changes. Few studies suggest that contaminant aqueous metals, like Cu or Ni at environmentally relevant concentrations, interfere with invertebrate chemical communication systems, such as that which operates between Daphnia and Chaoborus. The objective of our study was to determine if this interference could be also observed in lakes naturally rich in dissolved metals, such as volcanic lake (Lago Grande di Monticchio). This study aimed to assess if natural dissolved metals (e.g., Fe, Mn and Sr) could impair the ability of Daphnia pulex and D. galeata × hyalina × cucullata 'complex' populations to respond to Chaoborus kairomones by producing morphological defenses against potential predation, and to understand how Chaoborus predation might affect zooplankton community composition and overall zooplankton density. The predator impact did not result in: i) any morphological changes; ii) any apparent shift in body size pattern of the prey population; iii) any shift in life history traits. Chaoborus accounted for high mortality rates in Cladocera and strongly reduced the chance of individuals to reach maturity. Moreover, highly significant negative correlations between abundance of dominant taxa of zooplankton and C. flavicans were found. The last larval instars of C. flavicans seem to reduce the number of crustaceans, particularly cladocerans and copepod adults and could play an important role in structuring zooplankton communities. Our results suggest that metal inhibition of defence strategies induction probably occurs along the signal transduction pathway in Lake Grande di Monticchio. Impairment of chemosensory response to predatory chemical cues may have widespread ecological consequences in aquatic systems. Chaoborus predation effects can greatly affect both zooplankton biomass and community composition, impact interactions at lower trophic levels and generate an ecological cascade leading to a rapid eutrophication.

Scent of a Fly

Sexual courtship is a highly ritualized behavior in many animals. Recent work in the vinegar fly, Drosophila melanogaster, has illuminated how the pheromone cis-vaccenyl acetate modulates sexual behavior in the fly. Chemosensory receptors and a sexually dimorphic circuit activated by this pheromone have been identified. This minireview highlights recent advances in the field of fly courtship.

Use of pheromone mimic to cause the disintegration and collapse of colonies of tent caterpillars (Malacosoma spp.)

AbstractThe disruption of the trail‐based communication system of eastern tent caterpillars (ETC) (Malacosoma americanum) and forest tent caterpillars (FTC) (M. disstria) by spraying host trees with a trail pheromone mimic caused the disintegration and collapse of young colonies. A 1‐ppm formulation of 5β‐cholestan‐3‐one or a control formulation was sprayed on trees either before or after eclosion of the caterpillars in the spring. For pre‐eclosion trials, an average of 92% of the FTC colonies on control trees, but only 17% of colonies on treated trees consisted of viable, cohesive aggregates when survival was assessed 5–23 days post‐spray. The remaining colonies had either completely disappeared or were reduced to small, disintegrated fragments. For the ETC, 100% of colonies on control trees and an average of 12% of colonies on treated trees consisted of cohesive aggregates inhabiting tents when survival was assessed 3–41 days post‐spray. The remaining colonies were completely destroyed, reduced to scattered individuals, or to small, disintegrated, shelter‐less fragments. For post‐eclosion trials with the FTC in which colonies were sprayed during their first or second larval stadium, an average of 63% of individuals in control groups but only 7% in treatment groups were still on the sprayed trees 5–18 days post‐spray. For the ETC, colonies were largely unaffected when trees were not sprayed until after the caterpillars had eclosed and established tents. For all trials, the loss of individuals on treated trees was largely attributable to the inability of dispersed and isolated first or second instar caterpillars to maintain secure purchase, causing them to fall from trees and perish. To our knowledge, this is the first study to investigate the potential of manipulating populations of pest species by interfering with a trail‐based, chemical communication system.

Multiple functions of an odorant-binding protein in the mosquito Aedes aegypti

To prevent spreading of deadly diseases, populations of mosquitoes can be controlled by interfering with their chemical communication system. Odorant-binding proteins, recently shown to be required for olfaction, represent interesting targets for such purpose. Here we describe the ligand-binding properties and the unusual tissue expression of odorant-binding protein 22 from the repertoire of Aedes aegypti. Best ligands are molecules with two aromatic rings connected by a short rigid chain. The protein is expressed not only in sensory organs, such as the antennae and proboscis, but also in the male reproductive apparatus and transferred to the spermathecs of females. This suggests an additional function for this protein as pheromone carrier, analogously to vertebrates’ urinary and salivary proteins as well as some insect chemosensory proteins. Antiserum against odorant-binding protein 22 also stained the edges and sensilla of spiracles, indicating a third, still unknown, role for this protein.

Australian tadpoles do not avoid chemical cues from invasive cane toads (Bufo marinus)

If invasive species are phylogenetically distinct from native taxa, divergence in intraspecific communication systems may allow control via methods that invoke behavioural responses by the invasives but not by local species. Previous work has exploited sexual signals in this respect (e.g. species-specific mate-attraction pheromones) but there is equal potential to exploit non-sexual signals, such as chemically mediated behavioural responses of anuran larvae. Cane toads (Bufo marinus), originally from Central and South America, are creating major ecological problems during their invasion through Australia. In an earlier study, we showed that cane toad tadpoles are strongly repulsed by chemical cues from crushed conspecifics, suggesting that these animals possess significant chemical communication systems. To be useful in control of toads, such a response would need to be restricted to cane toads rather than all anurans. In laboratory trials, we detected only minor behavioural responses of six native Australian anuran species to chemical cues from cane toads. Native tadpoles (both hylids and myobatrachids) either ignored the stimulus, or tended to approach it rather than to avoid it. These results are encouraging for the potential use of toad-specific chemicals to manipulate the behaviour of tadpoles in the field, with few collateral effects on native Australian anurans.