Abstract
Interspecific relationships among insects are often mediated by chemical cues, including non-volatile cuticular compounds. Most of these compounds are hydrocarbons that necessitate the use of solvents for their extraction, identification, and manipulation during behavioral assays. The toxicity of these solvents often precludes the removal and reapplication of hydrocarbons from and to live insects. As a consequence, dummies often are used in behavioral assays, but their passivity can bias the behavior of the responding insects. To overcome these limitations, we propose a method where cuticular compounds are extracted from live ants by placing them into glass vials half-filled with tepid water (ca. 34°C) and vigorously shaking the vials to form an emulsion whose supernatant can be analyzed and/or reapplied to other ants. We demonstrate that cuticular compounds can be extracted from workers of the red fire ant, Solenopsis saevissima, and reapplied to the cuticle of workers from a sympatric species, Camponotus blandus (both Hymenoptera: Formicidae), while keeping the ants alive. Gas chromatographic-mass spectrometric analysis and behavioral assays were used to confirm the successful transfer of the behaviorally active compounds.
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References
Akino, T., Yamamura, K., Wakamura, S., and Yamaoka, R. 2004. Direct behavioral evidence for hydrocarbons as nestmate recognition cues in Formica japonica (Hymenoptera: Formicidae). Appl. Entomol. Zool. 39:381–387.
Andrews, E. A. 1911. Observation on termites in Jamaica. J. Anim. Behav. 1:193–228.
Bagnères, A. G. and Morgan, E. D. 1990. A simple method for analysis of insect cuticular hydrocarbons. J. Chem. Ecol. 16:3263–3276.
Blum, M. S., Murray, S., Walker, J. R., Callahan, P. S., and Novak, A. F. 1958. Chemical, insecticidal, and antibiotic properties of fire ant venom. Science 128:306–307.
Brand, J. M., Blum, M. S., Fales, H. M., and MacConnell, J. G. 1972. Fire ant venoms: comparative analyses of alkaloidal components. Toxicon 10:259–271.
Chen, L. and Fadamiro, H. Y. 2009a. Re-investigation of venom chemistry of Solenopsis fire ants. I. Identification of novel alkaloids in S. richteri. Toxicon 53:469–478.
Chen, L. and Fadamiro, H. Y. 2009b. Re-investigation of venom chemistry of Solenopsis fire ants. II. Identification of novel alkaloids in S. invicta. Toxicon 53:479–486.
Colazza, S., Aquila, G., De Pasquale, C., Peri, E., and Millar, J. 2007. The egg parasitoid Trissolcus basalis uses n-nonadecane, a cuticular hydrocarbon from its stink bug host Nezara viridula, to discriminate between female and male hosts. J. Chem. Ecol. 33:1405–1420.
Dani, F. R., Jones, G. R., Corsi, S., Beard, R., Pradella, D., and Turillazzi, S. 2005. Nestmate recognition cues in the honey bee: differential importance of cuticular alkanes and alkenes. Chem. Senses 30:477–489.
Dejean, A., Corbara, B., and Oliva-Rivera, J. 1990. Mise en évidence d’une forme d’apprentissage dans le comportement de capture des proies chez Pachycondyla (=Neoponera) villosa (Formicidae, Ponerinae). Behaviour 115:175–187.
Fresneau, D. 1980. Fermeture des sociétés et marquage territorial chez les fourmis ponérines du genre Neoponera. Biol Ecol Méditerr 7:205–206.
Gamboa, G. J., Reeve, H. K., and Holmes, W. G. 1991. Conceptual issues and methodology in kin recognition research, a critical discussion. Ethol. 88:109–127.
Haight, K. L. 2006. Defensiveness of the fire ant, Solenopsis invicta, is increased during colony rafting. Insect. Soc. 53:32–36.
Henderson, G., Andersen, J. F., Phillips, J. K., and Jeanne, R. L. 1990. Internest aggression and identification of possible nestmate discrimination pheromones in polygynous ant Formica montana. J. Chem. Ecol. 16:2217–2228.
Hölldobler, B. and Wilson, B. 1990. The ants. Harvard University Press, Cambridge
Holway, D. A., Lach, L., Suarez, A. V., Tsutsui, N. D., and Case, T. J. 2002. The causes and consequences of ant invasions. Annu. Rev. Ecol. Syst. 33:181–233.
Lenoir, A., D’Ettorre, P., Errard, C., and Hefetz, A. 2001. Chemical ecology and social parasitism in ants. Annu. Rev. Entomol. 46:573–599.
MacConnell, J. G., Blum, M. S., and Fales, H. M. 1971. The chemistry of fire ant venom. Tetrahedron 27:1129–1139.
Monnin, T., Malosse, C., and Peeters, C. 1998. Solid-phase microextraction and cuticular hydrocarbon differences related to reproductive activity in queenless ant Dinoponera quadriceps. J. Chem. Ecol. 24:473–490.
Morgan, E. D. 1990. Preparation of small samples for chromatography of insect pheromones. Anal. Chim. Acta 236:227–235.
Morrison, L. W. 1998. A review of Bahamian ant (Hymenoptera: Formicidae) biogeography. J. Biogeography 25:561–571.
Nelson, D. R., Fatland, C. L., Howard, R. W., McDaniel, C. A., and Blomquist, G. J. 1980. Re-analysis of the cuticular methylalkanes of Solenopsis invicta and S. richteri. Insect Biochem. 10:409–418.
Obin, M. S. and Vander Meer, R. K. 1985. Gaster flagging by fire ants (Solenopsis spp.): functional significance of venom dispersal behavior. J. Chem. Ecol. 11:1757–1768.
Orivel, J. and Dejean, A. 2002. Ant activity rhythms in a pioneer vegetal formation of French Guiana (Hymenoptera: Formicidae). Sociobiology 39:65–76.
Roulston, T. H., Buczkowski, G., and Silverman, J. 2003. Nestmate discrimination in ants: effect of bioassay on aggressive behavior. Insect. Soc. 50:151–159.
Roux, O., Gers, C., Tene-Ghomsi, J. N., Arvanitakis, L., Bordat, D., and Legal, L. 2007. Chemical characterization of contact semiochemicals for host-recognition and host-acceptance by the specialist parasitoid Cotesia plutellae (Kurdjumov). Chemoecol. 17:13–18.
Ruther, J., Sieben, S., and Schricker, B. 1998. Role of cuticular lipids in nestmate recognition of the European hornet Vespa crabro L. (Hymenoptera, Vespidae). Insect Soc. 45:169–179.
Ruther, J., Sieben, S., and Schricker, B. 2002. Nestmate recognition in social wasps: manipulation of hydrocarbon profiles induces aggression in the European hornet. Naturwissenschaften 89:111–114.
Tentschert, J., Bestmann, H. J., and Heinze, J. 2002. Cuticular compounds of workers and queens in two Leptothorax ant species—a comparison of results obtained by solvent extraction, solid sampling, and SPME. Chemoecol. 12:15–21.
Turillazzi, S., Sledge, M. F., and Moneti, G. 1998. Use of a simple method for sampling cuticular hydrocarbons from live social wasps. Ethol. Ecol. Evol. 10:293–297.
Vander Meer, R. K. and Morel, L. 1998. Nestmate recognition in ants, pp. 79–103, in R. K. Vander Meer, M. D. Breed, K. E. Espelie, and M. L. Winston (eds.). Pheromone communication in social insects: Ants, wasps, bees, and termites. Westview, Boulder.
Wagner, D., Tissot, M., Cuevas, W., and Gordon, D. M. 2000. Harvester ants utilize cuticular hydrocarbons in nestmate recognition. J. Chem. Ecol. 26:2245–2257.
Acknowledgments
We are grateful to Andrea Dejean for proofreading the manuscript, to Antoine Steven from the Institut Pasteur de la Guyane for allowing us the use of a GC-MS, to Felipe Ramon-Portugal for technical assistance, and to the staff of the Laboratoire Environnement de Petit Saut for field accommodations. Two anonymous reviewers are thanked for their helpful suggestions, which significantly improved the manuscript. Financial support for this study was provided by the Programme Amazonie II of the French Centre National de la Recherche Scientifique (project 2ID) and the Programme Convergence 2007–2013, Région Guyane from the European Community (project DEGA).
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Roux, O., Martin, JM., Ghomsi, N.T. et al. A Non-lethal Water-based Removal-reapplication Technique for Behavioral Analysis of Cuticular Compounds of Ants. J Chem Ecol 35, 904–912 (2009). https://doi.org/10.1007/s10886-009-9673-x
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DOI: https://doi.org/10.1007/s10886-009-9673-x