Eusocial insects use cuticular hydrocarbons as components of pheromones that mediate social behaviours, such as caste and nestmate recognition, and regulation of reproduction. In ants such as Harpegnathos saltator, the queen produces a pheromone which suppresses the development of workers’ ovaries and if she is removed, workers can transition to a reproductive state known as gamergate. Here we functionally characterize a subfamily of odorant receptors (Ors) with a nine-exon gene structure that have undergone a massive expansion in ants and other eusocial insects. We deorphanize 22 representative members and find they can detect cuticular hydrocarbons from different ant castes, with one (HsOr263) that responds strongly to gamergate extract and a candidate queen pheromone component. After systematic testing with a diverse panel of hydrocarbons, we find that most Harpegnathos saltator Ors are narrowly tuned, suggesting that several receptors must contribute to detection and discrimination of different cuticular hydrocarbons important in mediating eusocial behaviour.

The endoparasitic wasps (Hymenoptera: Braconidae), Microplitis croceipes (specialist), and Cotesia marginiventris (generalist) are parasitoids of lepidopteran larvae that differ in their degree of host specificity. Previous studies by our group have reported key differences in the response of both parasitoids to host-related volatiles. To possibly understand the basis for our results, we compared the abundance of antennal sensilla in both sexes of both parasitoid species using scanning electron microscopy and silver-staining techniques. Males of both species had relatively longer antennae than conspecific females. Five major sensilla types were recorded in both species: sensilla chaetica (nonporous), s. trichodea (nonporous), s. placodea (multiporous), s. basiconica (two types, type 1 with terminal opening and type 2 with wall pores), and s. coeloconica (nonporous). Silver staining confirmed multiple pores on the walls of s. placodea and s. basiconica type 2 and terminal openings on s. basiconica type 1, suggesting chemosensory function. In general, both putative chemosensilla types, s. placodea and s. basiconica, were more abundant in M. croceipes (specialist) than in C. marginiventris (generalist), and this was true for both sexes. Comparing the sexes, s. placodea and s. trichodea were significantly more abundant in M. croceipes males compared to females. In contrast, s. placodea was relatively more abundant in female C. marginiventris than in males. These results may explain the reported differences in the responses of both parasitoid species to host-related volatiles and are discussed in relation to the possible roles of the sensilla types in the behavioral ecology of the parasitoids.

The endoparasitic wasps (Hymenoptera: Braconidae), Microplitis croceipes (specialist) and Cotesia marginiventris (generalist), are parasitoids of lepidopteran larvae and differ in their degree of host specificity. Recent studies have reported key differences between the two species in the abundance of antennal olfactory sensilla and their response to host-related volatiles. Here, we have compared antennal lobe architecture and glomerular organization in the two parasitoid species by using a combination of axonal tract tracing techniques and confocal microscopy. In M. croceipes, the medial half of the antennal lobe is larger with a greater number of glomeruli compared with the lateral half, whereas in C. marginiventris, the lateral half is larger than the median half. The volume of the antennal lobe is approximately 2.5 times greater in M. croceipes than in C. marginiventris. However, the number of glomeruli per antennal lobe is only slightly higher in M. croceipes (females: 219-222; males: 220-224) than in C. marginiventris (females: 192-194; males: 193-196). A comparison of males and females within each species demonstrated a striking sexual difference in terms of an enlarged glomerulus (macroglomerulus or MG) at the entrance of the antennal nerve and of a complex of 3-4 MG (CMG) in the posterior region of the antennal lobe of males of both species. Being specific to males, both the MG and CMG might be involved in the detection of female-related odor.

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Highlights A novel geotaxis assay showed high intensity odorant exposures are harmful to flies  Repulsion at high odor intensities can be a protective mechanism  Olfactory receptor neuron (ORN) excitability abruptly changes with odor intensity  A linear combination of ORN activities can robustly predict intensity-dependent behavioral repulsion SummaryThe olfactory system is uniquely positioned to warn an organism of environmental threats. Whether and how it encodes such information is not understood. Here, we examined this issue in the fruit fly Drosophila melanogaster. We found that intensitydependent repulsion to chemicals safeguarded flies from harmful, high-intensity vapor exposures. To understand how sensory input changed as the odor valence switched from innocuous to threatening, we recorded from olfactory receptor neurons (ORNs) in the fly antenna. Primarily, we observed two response non-linearities: recruitment of nonactive ORNs at higher intensities, and abrupt transitions in neural excitability from regular spiking to high-firing oscillatory regime. Although non-linearities observed in any single ORN was not a good indicator, a simple linear combination of firing events from multiple neurons provided robust recognition of threating/repulsive olfactory stimuli. In sum, our results reveal how information necessary to avoid environmental threats may also be encoded in the insect antenna.