Cuticular Hydrocarbon Profile of Parasitic Beetles, Aethina tumida (Coleoptera: Nitidulidae)

Cuticular hydrocarbons (CHCs) cover insects’ bodies and play important roles in chemical communication, including nestmate recognition, for social insects. To enter colonies of a social host species, parasites may acquire host-specific CHCs or covertly maintain their own CHC profile by lowering its quantity. However, the chemical profile of small hive beetles (SHBs), Aethina tumida, which are parasites of honey bee, Apis mellifera, colonies, and other bee nests, is currently unknown. Here, adults of SHB and honey bee host workers were collected from the same field colonies and their CHC profiles were analysed using GC-MS. The chemical profiles of field-sampled SHBs were also compared with those of host-naive beetles reared in the laboratory. Laboratory-reared SHBs differed in their CHC profiles from field-sampled ones, which showed a more similar, but ten-fold lower, generic host CHC profile compared to host workers. While the data confirm colony-specific CHCs of honey bee workers, the profile of field-collected SHBs was not colony-specific. Adult SHBs often commute between different host colonies, thereby possibly preventing the acquisition of a colony-specific CHC profiles. An ester was exclusive to both groups of SHBs and might constitute an intraspecific recognition cue. Our data suggest that SHBs do not use any finely tuned chemical strategy to conceal their presence inside host colonies and instead probably rely on their hard exoskeleton and defence behaviours.

Social signaling via bioluminescent blinks determines nearest neighbor distance in schools of flashlight fish Anomalops katoptron

The schooling flashlight fish Anomalops katoptron can be found at dark nights at the water surface in the Indo-Pacific. Schools are characterized by bioluminescent blink patterns of sub-ocular light organs densely-packed with bioluminescent, symbiotic bacteria. Here we analyzed how blink patterns of A. katoptron are used in social interactions. We demonstrate that isolated specimen of A. katoptron showed a high motivation to align with fixed or moving artificial light organs in an experimental tank. This intraspecific recognition of A. katoptron is mediated by blinking light and not the body shape. In addition, A. katoptron adjusts its blinking frequencies according to the light intensities. LED pulse frequencies determine the swimming speed and the blink frequency response of A. katoptron, which is modified by light organ occlusion and not exposure. In the natural environment A. katoptron is changing its blink frequencies and nearest neighbor distance in a context specific manner. Blink frequencies are also modified by changes in the occlusion time and are increased from day to night and during avoidance behavior, while group cohesion is higher with increasing blink frequencies. Our results suggest that specific blink patterns in schooling flashlight fish A. katoptron define nearest neighbor distance and determine intraspecific communication.

Analysis of cuticular chemical profiles of Latrodectus geometricus (Araneae: Theridiidae) females and juveniles using GC×GC/qMS

Communication in spiders can occur by several mechanisms, such as chemical cues and the cuticular hydrocarbons (CHCs) play an important role in intraspecific recognition. Several techniques have been used to evaluate CHCs in spiders, such as Gas Chromatography with Flame Ionization Detection, Gas Chromatography coupled with Mass Spectrometric detection and recently Fourier Transformed Infrared Photoacoustic Spectroscopy. In this study, rapid-scanning two-dimensional gas chromatography with quadrupole mass spectrometry (GC×GC/qMS) was employed to assess the CHCs of L. geometricus females and juveniles of different ages. The results demonstrate that there is variation in CHCs of different body parts of females, with abdomen presenting greater number of compounds, as well as between adults and juveniles. Branched alkanes represented the majority of compounds in all samples, followed by linear alkanes, and alkenes were only present in adult’s abdomen and cephalothorax. The compounds 10-methyloctacosaneand 14-methyloctacosane; and 10-methyltriacontane and 14-methyltriacontane showed the same retention time in the 1D and were separated in the 2D. According to the results, rapid-scanning GC×GC/qMS can be considered a reliable technique to assess CHCs since it was possible to identify the spider’s cuticular compounds, and detect and separate two cases of chromatographic coelution.

Intraspecific Variation of the Composition of Linear Alkanes in Social Wasp Mischocyttarus consimilis

An important attribute of the evolution and maintenance of sociality in insects is their ability to distinguish members of their own colonies by means of chemical signals during their interactions. Of this type of signals, the cuticular hydrocarbons, responsible among other functions, for intraspecific recognition stand out. Linear alkanes are indicated as the class of compounds that would be most involved in water retention in the body of insects, however, some studies have investigated their role as mediators of interactions. Thus it is possible that there is significant intraspecific variation of its composition, so the objective of this study was to test the hypothesis that linear alkanes vary significantly among populations, castes and sexes, as well as between newly emerged females of different ages of the Mischocyttarus consimilis Zikán 1949 wasp. The samples were analyzed by gas chromatography with flame ionization detector and gas chromatography coupled to mass spectrometry. The results show that there is a significant variation between the chemical cuticular profiles of samples from different populations, as well as between castes, sex and newly emergent workers of different ages. Therefore, it is possible to infer that this class of compounds may vary according to genetic differences between populations, but also by different environmental conditions. The differences between castes, sex and ages suggest that these compounds may also be involved in mediating interactions between nestmates.

Linear Alkanes and Reproductive Status of Polistes versicolor (Hymenoptera: Vespidae) Females in Winter Aggregates

Female wasps such as Polistes versicolor can form aggregates to face weather conditions that are not suitable to sustain their colonies. The interactions between individuals in these aggregates, just as in other associations, are probably facilitated by chemical signals. Of these compounds some of the most efficient during social interactions of insects are those called contact pheromones or superficial pheromones. This special type of pheromones, known as cuticular hydrocarbons, can be found in insects cuticle. They facilitate the differentiation of caste, species and nestmates, and may be important indicators of dominance as well as fertility. Some studies indicate that linear alkanes are important cuticular compounds for intraspecific recognition and discrimination. The aim of this study was to evaluate the relationship between reproductive physiologic condition and the linear alkanes present in the cuticle of females of P. versicolor in aggregates employing Gas Chromatography with Flame Ionization Detector (GC-FID). Females from distinct aggregates were differentiated by the chemical composition of their cuticle. In each aggregate, there was difference in cuticular chemical composition between females with different ovarian development degrees, allowing the distinction between inseminated and non-inseminated females.

Photo-identification as a technique for recognition of individual fish: a test with the freshwater armored catfish Rineloricaria aequalicuspis Reis & Cardoso, 2001 (Siluriformes: Loricariidae)

Photo-identification allows individual recognition of animal species based on natural marks, being an alternative to other more stressful artificial tagging/marking techniques. An increasing number of studies with different animal groups has shown that photo-identification can successfully be used in several situations, but its feasibility to study freshwater fishes is yet to be explored. We demonstrate the potential use of photo-identification for intraspecific recognition of individuals in the stream-dwelling loricariid Rineloricaria aequalicuspis . We tested photo-identification in laboratory and field conditions based on the interindividual variability in abdominal bony plates. Our test yielded high correct matches in both laboratory (100%) and field conditions (> 97%), comparable to other reliable techniques and to studies that successfully used photo-identification in other animals. In field conditions, the number of correct matches did not differ statistically between computer-assisted and naked-eye identification. However, the average time expended to conclude computer-assisted photo evaluations was about half of the time expended to conclude naked-eye evaluations. This result may be exacerbated when using database with large number of images. Our results indicate that photo-identification can be a feasible alternative technique to study freshwater fish species, allowing for a wider use of mark-recapture in ecological and behavioral studies.

Polarized iridescence of the multilayered elytra of the Japanese jewel beetle, Chrysochroa fulgidissima

The elytra of the Japanese jewel beetle Chrysochroa fulgidissima are metallic green with purple stripes. Scanning electron microscopy and atomic force microscopy demonstrated that the elytral surface is approximately flat. The accordingly specular green and purple areas have, with normal illumination, 100–150 nm broad reflectance bands, peaking at about 530 and 700 nm. The bands shift progressively towards shorter wavelengths with increasing oblique illumination, and the reflection then becomes highly polarized. Transmission electron microscopy revealed that the epicuticle of the green and purple areas consists of stacks of 16 and 12 layers, respectively. Assuming gradient refractive index values of the layers between 1.6 and 1.7 and applying the classical multilayer theory allowed modelling of the measured polarization- and angle-dependent reflectance spectra. The extreme polarized iridescence exhibited by the elytra of the jewel beetle may have a function in intraspecific recognition.

The genetics of mating recognition between Drosophila simulans and D. sechellia

During courtship, visual and chemical signals are often exchanged between the sexes. The proper exchange of such signals ensures intraspecific recognition. We have examined the genetic basis of interspecific differences in male mating behaviour and pheromone concentration between Drosophila simulans and D. sechellia by using Drosophila simulans/D. sechellia introgression lines. Our results show a majority of quantitative trait loci (QTLs) explaining variation in both male mating behaviour and pheromone concentration to be located on the third chromosome. One QTL found on the third chromosome explains variation in time needed to start courtship and copulation as well as time spent courting. The position of such QTL (approximately 84A–88B) with effects on courtship and copulation aspects of mating includes the candidate sex determination gene doublesex (84E5–6) and Voila (86E1–2), a gene that affects male courtship in D. melanogaster. One additional third chromosome QTL explained variation in 7-tricosene pheromone concentrations among males. The interval mapping position of this QTL (approximately 68E–76E) did not overlap with the position detected for differences in mating behaviour and the intervals did not include candidate genes previously identified as having an effect on D. melanogaster cuticular hydrocarbon production. We did not detect any directionality of the effect of Drosophila sechellia allele introgressions in male mating recognition.