The present paper shows, by means of single-cell recordings, responses of antennal sensory neurons of the damselfly Ischnura elegans when stimulated by air streams at different CO2 concentrations. Unlike most insects, but similarly to termites, centipedes and ticks, Odonata possess sensory neurons strongly inhibited by CO2, with the magnitude of the off-response depending upon the CO2 concentration. The Odonata antennal sensory neurons responding to CO2 are also sensitive to airborne odors; in particular, the impulse frequency is increased by isoamylamine and decreased by heptanoic and pentanoic acid. Further behavioral investigations are necessary to assign a biological role to carbon dioxide detection in Odonata.

Heritable microbes are an important component of invertebrate biology, acting both as beneficial symbionts and reproductive parasites. Whilst most previous research has focussed on the 'Wolbachia pandemic', recent work has emphasised the importance of other microbial symbionts. In this study, we present a survey of odonates (dragonflies and damselflies) for torix group Rickettsia, following previous research indicating that this clade can be common in other aquatic insect groups. PCR assays were used to screen a broad range of odonates from two continents and revealed 8 of 76 species tested were infected with Rickettsia. We then conducted further deeper screening of UK representatives of the Coenagrionidae damselfly family, revealing 6 of 8 UK coenagrionid species to be positive for torix Rickettsia. Analysis of Rickettsia gene sequences supported multiple establishments of symbiosis in the group. Some strains were shared between UK coenagrionid species that shared mtDNA barcodes, indicating a likely route for mitochondrial introgression between sister species. There was also evidence of coinfecting Rickettsia strains in two species. FISH analysis indicated Rickettsia were observed in the ovarioles, consistent with heritable symbiosis. We conclude that torix Rickettsia represent an important associate of odonates, being found in a broad range of species from both Europe and South America. There is evidence that coinfection can occur, vertical transmission is likely, and that symbiont movement following hybridisation may underpin the lack of 'barcoding gap' between well-established species pairs in the genus. Future work should establish the biological significance of the symbioses observed.

Here we describe the antennal lobe of Libellula depressa (Odonata, Libellulidae), identified on the basis of the projections of the afferent sensory neurons stemming from the antennal flagellum sensilla. Immunohistochemical neuropil staining as well as antennal backfills revealed sensory neuron terminal arborizations covering a large portion of the antennal lobe. No clear glomerular structure was identified, thus suggesting an aglomerular antennal lobe condition as previously reported in Palaeoptera. The terminal arbors of backfilled sensory neurons do, however, form spherical knots, probably representing the connections between the few afferent neurons and the antennal lobe interneurons. The reconstruction revealed that the proximal part of the antennal nerve is divided into two branches that innervate two spatially separated areas of the antennal lobe, an anterioventral lobe and a larger posteriodorsal lobe. Our data are consistent with the hypothesis that one tract of the antennal nerve of L. depressa contains olfactory sensory neurons projecting into one of the sublobes, while the other tract contains thermo-hygroreceptive neurons projecting into the other sublobe.

Climate change is expected to have substantial impacts on the composition of freshwater communities, and many species are threatened by the loss of climatically suitable habitat. In this study we identify Australian Odonata (dragonflies and damselflies) vulnerable to the effects of climate change on the basis of exposure, sensitivity and pressure to disperse in the future. We used an ensemble of species distribution models to predict the distribution of 270 (85%) species of Australian Odonata, continent-wide at the subcatchment scale, and for both current and future climates using two emissions scenarios each for 2055 and 2085. Exposure was scored according to the departure of temperature, precipitation and hydrology from current conditions. Sensitivity accounted for change in the area and suitability of projected climatic habitat, and pressure to disperse combined measurements of average habitat shifts and the loss experienced with lower dispersal rates. Streams and rivers important to future conservation efforts were identified based on the sensitivity-weighted sum of habitat suitability for the most vulnerable species. The overall extent of suitable habitat declined for 56-69% of the species modelled by 2085 depending on emissions scenario. The proportion of species at risk across all components (exposure, sensitivity, pressure to disperse) varied between 7 and 17% from 2055 to 2085 and a further 3-17% of species were also projected to be at high risk due to declines that did not require range shifts. If dispersal to Tasmania was limited, many south-eastern species are at significantly increased risk. Conservation efforts will need to focus on creating and preserving freshwater refugia as part of a broader conservation strategy that improves connectivity and promotes adaptive range shifts. The significant predicted shifts in suitable habitat could potentially exceed the dispersal capacity of Odonata and highlights the challenge faced by other freshwater species.

Selection for the oviposition site represents the criterion for the behavioral process of habitat selection for the next generation. It is well known that in Odonata the most general cues are detected visually, but laboratory investigations on the coenagrionid Ischnura elegans showed through behavioral and electrophysiological assays that adults were attracted by olfactory cues emitted by prey and that males of the same species are attracted by female odor. The results of the present behavioral and electrophysiological investigations on I. elegans suggest the involvement of antennal olfactory sensilla in oviposition behavior. In particular, I. elegans females laid in the laboratory significantly more eggs in water from larval rearing aquaria than in distilled or tap water. Moreover, the lack of preference between rearing water and tap water with plankton suggests a role of volatiles related to conspecific and plankton presence in the oviposition site choice. I. elegans may rely on food odor for oviposition site selection, thus supporting the predictions of the "mother knows best" theory. These behavioral data are partially supported by electroantennographic responses. These findings confirm a possible role of olfaction in crucial aspects of Odonata biology.

Perfluorinated substances are global and ubiquitous pollutants. However, very little is known about these substances in invertebrates, and even less in terrestrial invertebrates in particular. We analysed adult male dragonflies from six sites in South Africa for perfluoroalkyl substances (PFASs), including perfluorooctanesulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluro-n-undecanoic acid (PFUnA), perfluoro-n-dodecanoic acid (PFDoA), perfluorohexanoic acid (PFHxA), and perfluorohexane sulfonic acid (PFHxS). PFOS was detected in all individuals, with less quantifiable occurrences of the other substances. The dragonflies from the three northern sites located in farming areas had significantly lower ΣPFASs concentrations than the southern sites located closer to industrial areas (median ΣPFASs of 0.32 ng/g wm (wet mass) for North, and 9.3 ng/g wm for South). All substances except PFOS occurred at similar concentrations at all six sites when quantifiable, but PFOS dominated in the Southern sites. The highest median concentration was from Bloemhof Dam (ΣPFASs = 21 ng/g wm), which is known to be polluted by PFOS. Perfluorinated substances are not known to be manufactured in South Africa, therefore the residues detected are likely to have been derived from imported products. Odonata play a significant role in freshwater ecology. Any impacts on these aquatic and aerial predators are likely to have effects on aquatic and associated ecosystems. Further studies are required over a much larger geographic region and to investigate sources.

All natural populations show fluctuations in space or time. This is fundamental for the maintenance of biodiversity, as it allows species to coexist. Long-term ecological studies are rare, mainly due to logistics, but studies like the one presented below recognize the dimensionality of temporal change and the ecological processes that lead to shifts in community composition over time. Here, we used three sampling occasions from a dataset spanning 20 years where dragonflies in central Sweden were monitored. Our aim was to investigate how the prevalence of ecological and biological species traits varied over time measured as Community-level Weighted Means of trait values (CWM). Most CWM values varied significantly between years. Most of the traits changed between the second and the last sampling occasion, but not between the two first ones. These changes could be linked to major changes in species abundance. Our work indicates that fundamental shifts in community structure can occur over a short time, providing environmental drivers act on species turnover. In our case, Climate change and pH levels in lakes are most likely the most important factors.

Wolbachia is one of the most common endosymbionts found infecting arthropods. Theory predicts symbionts like Wolbachia will be more common in species radiations, as host shift events occur with greatest frequency between closely related species. Further, the presence of Wolbachia itself may engender reproductive isolation, and promote speciation of their hosts. Here we screened 178 individuals belonging to 30 species of the damselfly genera Nesobasis and Melanesobasis - species radiations endemic to the Fiji archipelago in the South Pacific - for Wolbachia, using multilocus sequence typing to characterize bacterial strains. Incidence of Wolbachia was 71% in Nesobasis and 40% in Melanesobasis, and prevalence was also high, with an average of 88% in the Nesobasis species screened. We identified a total of 25 Wolbachia strains, belonging to supergroups A, B and F, with some epidemic strains present in multiple species. The occurrence of Wolbachia in both males and females, and the similar global prevalence found in both sexes rules out any strong effect of Wolbachia on the primary sex-ratio, but are compatible with the phenotype of cytoplasmic incompatibility. Nesobasis has higher species richness than most endemic island damselfly genera, and we discuss the potential for endosymbiont-mediated speciation within this group.

Odonata can be sampled following different types of protocols. In Brazil, the most used protocol is the scanning in fixed areas method, where a 100-meter transect is delimited in one of the stream margins, subdivided into 20 segments measuring 5 meters. Despite being universally used, the methodological efficiency or limitations of this protocol for Odonata has never been tested. In this scenario, our objective was to assess the efficiency of the sampling protocol to measure the richness and composition of Odonata in three fundamental aspects: the time of sampling and sampling effort over time and space. We show that the best sampling efficiency was achieved in collections performed at noon, in transects measuring 100 meters, requiring at least two samplings in the same location, supporting the procedures traditionally adopted by many studies with the group. While comparing species composition, we did not see any implication between the different treatments on the capture of the local species pool. However, we highlight and discuss some possible methodological flaws when using this protocol to sample specific Odonata groups. We believe the results obtained are fundamental in the inventory of species and to conduct future studies, as well as to aid conservative measures that use the order Odonata as a tool for environmental monitoring.

Odonata (dragonflies and damselflies) present an unparalleled insect model to integrate evolutionary genomics with ecology for the study of insect evolution. Key features of Odonata include their ancient phylogenetic position, extensive phenotypic and ecological diversity, several unique evolutionary innovations, ease of study in the wild and usefulness as bioindicators for freshwater ecosystems worldwide. In this review, we synthesize studies on the evolution, ecology and physiology of odonates, highlighting those areas where the integration of ecology with genomics would yield significant insights into the evolutionary processes that would not be gained easily by working on other animal groups. We argue that the unique features of this group combined with their complex life cycle, flight behaviour, diversity in ecological niches and their sensitivity to anthropogenic change make odonates a promising and fruitful taxon for genomics focused research. Future areas of research that deserve increased attention are also briefly outlined.

Among the winged insects (Pterygota) the Dragonflies and Damselflies (Odonata) are unique for several reasons. Behaviourally they are aerial predators that hunt and catch their prey in flight, only. Morphologically the flight apparatus of Odonata is significantly different from what is found in the remaining Pterygota. However, to understand the phylogenetic relationships of winged insects and the origin and evolution of insect flight in general, it is essential to know how the elements of the odonatan flight apparatus relate to those of the other Pterygota. Here we present a comprehensive, comparative morphological investigation of the thoracic flight musculature of damselflies (Zygoptera). Based on our new data we propose a homologization scheme for the thoracic musculature throughout Pterygota. The new homology hypotheses will allow for future comparative work and especially for phylogenetic analyses using characters of the thoracic musculature throughout all winged insects. This will contribute to understand the early evolution of pterygote insects and their basal phylogenetic relationship.

Among the winged insects (Pterygota) the Odonata (dragon- and damselflies) are special for several reasons. They are strictly aerial predators showing remarkable flight abilities and their thorax morphology differs significantly from that of other Pterygota in terms of the arrangement and number of muscles. Even within one individual the musculature is significantly different between the nymphal and adult stage.

We identified and classified damselfly (Zygoptera) and dragonfly (Anisoptera) metacommunities in Brazilian Amazonia, relating species distribution patterns to known biological gradients and biogeographical history. We expected a random distribution of both Zygoptera and Anisoptera within interfluves. At the Amazonian scale, we expected Anisoptera metacommunities to be randomly distributed due to their higher dispersal ability and large environmental tolerance. In contrast, we expected Zygoptera communities to exhibit a Clementsian pattern, limited by the large Amazonia rivers due to their low dispersal ability.

Dragonflies and damselflies (Odonata) are important components in biomonitoring due to their amphibiotic lifecycle and specific habitat requirements. They are charismatic and popular insects, but can be challenging to identify despite large size and often distinct coloration, especially the immature stages. DNA-based assessment tools rely on validated DNA barcode reference libraries evaluated in a supraregional context to minimize taxonomic incongruence and identification mismatches.

The species of Epiophlebia are unique among the recent Odonata in showing a mixture of morphological characters of dragonflies (Anisoptera) and damselflies (Zygoptera). The status of the four described extant species of Epiophlebia is disputable from a genetic as well as from a morphological point of view. Here we present an analysis of the thoracic musculature of different nymphal instars of Epiophlebia laidlawi and Epiophlebia superstes to elucidate their morphology and ontogenetic development. In total, 75 muscles have been identified in the thorax of Epiophlebia. This represents the highest number of thoracic muscles ever found in any odonate. It includes six muscles that are reported for the first time for Odonata, and three of these are even new for Pterygota. In total, our results indicate that Epiophlebia has the most ancestral thoracic morphology among Odonata.

A dragonfly larva migrates from the water to the shore, perches on a plant stem and grasps it with strongly flexed legs. Adult legs inside the larval exoskeleton fit to the larval legs joint-to-joint. The adult emerges with stretched legs. During the molt, an imaginal leg must follow all the angles in exuvial joints. In turn, larval apodemes are withdrawn from imaginal legs. We visualized transient shapes of the imaginal legs by the instant fixation of insects at different moments of the molt, photographed isolated exuvial legs with the imaginal legs inside and then removed the exuvial sheath. Instant shapes of the imaginal tibia show sharp intrapodomere bends copying the angle in the larval femoro-tibial joint. The site of bending shifts distad during the molt. This is possible if the imaginal leg is pliable. The same problem of leg squeezing is also common in hemimetabolous insects as well as in other arthropods, whereas holometabolous insects overcome problems of a tight confinement either by using leg pliability in other ways but not squeezing (cyclorrhaphan flies, mosquitoes) or by pulling hardened legs out without change of their pupal zigzag configuration (moths, ants, honey bees). The pupal legs are not intended to grasp any external substrate.

Dragonflies and damselflies are among the earliest flying insects with extant representatives. However, unraveling details of their long evolutionary history, such as egg laying (oviposition) strategies, is impeded by unresolved phylogenetic relationships, particularly in damselflies. Here we present a transcriptome-based phylogenetic reconstruction of Odonata, analyzing 2,980 protein-coding genes in 105 species representing nearly all the order's families. All damselfly and most dragonfly families are recovered as monophyletic. Our data suggest a sister relationship between dragonfly families of Gomphidae and Petaluridae. According to our divergence time estimates, both crown-Zygoptera and -Anisoptera arose during the late Triassic. Egg-laying with a reduced ovipositor apparently evolved in dragonflies during the late Jurassic/early Cretaceous. Lastly, we also test the impact of fossil choice and placement, particularly, of the extinct fossil species, †Triassolestodes asiaticus, and †Proterogomphus renateae on divergence time estimates. We find placement of †Proterogomphus renateae to be much more impactful than †Triassolestodes asiaticus.

Ceriagrion fallax is ubiquitous in south China and is particularly easy be found in some rice fields. In this study, we sequenced and analyzed the complete mitochondrial genome (mitogenome) of C. fallax. This mitogenome was 15,350 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs) and two ribosomal RNA unit genes (rRNAs). The nucleotide composition of the mitogenome was biased toward A and T, with 74.0% of A + T content (A 42.1%, T 31.9%, C 14.6%, G 11.4%). Gene order was conserved and identical to most other previously sequenced Zygoptera dragonflies. Most PCGs of C. fallax have the conventional start codons ATN (seven ATG, two ATT, and two ATC), with the exception of nad3 and nad1 (TTG). Except for four PCGs (cox1, cox2, cox3, and nad5) end with the incomplete stop codon T--, all other PCGs terminated with the stop codon TAA. Phylogenetic analysis showed that C. fallax got together with the same family species (Agriocnemis femina, Enallagma cyathigerum, Ischnura elegans, Ischnura pumilio) with high support value. The relationships (Megapodagrionidae + ((Calopterygidae + (Euphaeidae + Pseudolestidae)) + (Coenagrionidae + Platycnemididae))) were supported within Zygoptera.

DNA barcoding has become a promising means for identifying organisms of all life stages. Currently, phenetic approaches and tree-building methods have been used to define species boundaries and discover 'cryptic species'. However, a universal threshold of genetic distance values to distinguish taxonomic groups cannot be determined. As an alternative, DNA barcoding approaches can be 'character based', whereby species are identified through the presence or absence of discrete nucleotide substitutions (character states) within a DNA sequence. We demonstrate the potential of character-based DNA barcodes by analysing 833 odonate specimens from 103 localities belonging to 64 species. A total of 54 species and 22 genera could be discriminated reliably through unique combinations of character states within only one mitochondrial gene region (NADH dehydrogenase 1). Character-based DNA barcodes were further successfully established at a population level discriminating seven population-specific entities out of a total of 19 populations belonging to three species. Thus, for the first time, DNA barcodes have been found to identify entities below the species level that may constitute separate conservation units or even species units. Our findings suggest that character-based DNA barcoding can be a rapid and reliable means for (i) the assignment of unknown specimens to a taxonomic group, (ii) the exploration of diagnosability of conservation units, and (iii) complementing taxonomic identification systems.

We present the first empirical treatment of the northernmost breeding dragonfly, Somatochlora sahlbergi. We sequenced populations from United States, Canada, Finland, Sweden and Norway for cytochrome oxidase I (COI) and D2 region of 28s. We found that, despite geographic barriers across its vast arctic range, S. sahlbergi is a single species. Not only does it appear to interbreed across its entire range, there also seems to be almost no variation among European and North American populations in their COI gene fragment (the barcode gene), which is usually extremely variable. We further found that characters thought to be diagnostic for the larvae of S. sahlbergi were absent in our European samples. We review and re-describe the habitat of this species based on new findings from recent field observations. Finally, we report for the first time the likely presence of this species in Japan. We hope our findings will encourage further study of this species and other under-studied insect taxa that inhabit the remote Arctic.