Male Damselflies Research Articles

Sex wars: a female genital spine forces male damselflies to shorten copulation duration.

In some species, males use weapons to harm females, increasing their short-term fitness. Here we show that females can use genital adaptations against males. Females of the damselfly Enallagma cyathigerum have a conspicuous vulvar spine on the eighth abdominal segment, which contacts with the male during copulation. We tested 3 hypotheses for its function: it (a) inflicts damage to the male during copulation; (b) facilitates endophytic oviposition; and (c) stimulates males during copulation to increase their investment. We found that males mated on average for 54 min with control females, but increased copulation to 99 min with females without spine. There was no evidence of physical harm of the spine on the male's seminal vesicle, which shows 8- to 18-folds, exactly where the spine contacts during copulation. Females with and without spine exhibited the same egg-laying rates and showed similar fecundity and fertility. Longevity was also similar in males mated to control and spineless females. In contrast to many species where females resist male harassment by behavioral responses, the morphological adaptation observed in E. cyathigerum appears to act as a sexual weapon, allowing females to control copulation duration. We suggest that the spine has evolved because of sexual conflict over mating duration.

Male agility in relation to mating success in two non‐territorial damselflies

AbstractMale body size is usually correlated with mating success in insects. For non‐territorial species, the small male advantage hypothesis predicts that smaller males may be favoured in sexual selection because small body size may predict agility and manoeuvrability. Consequently, selection for male size may drive the evolution of female‐biased sexual size dimorphism (SSD). Nevertheless, recent evidence shows that non‐territorial male damselflies that actively search for females may be larger and more agile than those that perch and wait for females to appear. Thus, here we addressed whether the male size is an indicator of male agility and if it may be used to predict male reproductive success in two non‐territorial damselflies. To this end, we conducted a field study with Acanthagrion truncatum Selys, 1876 and A. lancea Selys, 1876 (Zygoptera, Coenagrionidae). Males of A. truncatum adopt a sit‐and‐wait mating strategy, while the males actively search for females in A. lancea. Hence, we expected different selective forces acting on male body size and agility in these species. We compared the body size and wing length of mated and unmated males, and between sexes, to describe their patterns of SSD. Our results suggest that wing length can be used as a proxy for male body size and agility in both species. However, we have found no evidence for the small male advantage that could explain wing dimorphism in A. truncatum, nor selection for larger males in A. lancea. In conclusion, this study corroborates other studies that suggest agility cannot explain SSD in non‐territorial damselflies and fail to support the small male advantage hypothesis.

Adult damselflies as possible regulators of mosquito populations in urban areas.

Dragonfly and damselfly larvae have been considered as possible biocontrol agents against young instars of mosquito vectors in urban environments. Yet our knowledge about adult odonate predation against mosquito adults is scarce. We quantified daily and annual predation rates, consumption rates and prey preferences of adult Hetaerina vulnerata male damselflies in an urban park. A focus on predation of mosquito species was provided, quantified their arbovirus (dengue, chikungunya and Zika) infection rates and biting activity. Foraging times of H. vulnerata overlapped with those of the maximum activity of hematophagous mosquitoes. The most consumed preys were Diptera and Hymenoptera and, in lower quantities, Hemiptera, Coleoptera, Trichoptera, Psocoptera and Neuroptera. Of note, 7% of the diet was represented by hematophagous dipterans, with 2.4% being Aedes aegypti and Aedes albopictus. Prey abundance in the diet coincided with that of the same species in the environment. The arboviral infection rate (dengue, chikungunya and Zika) was 1.6% for A. aegypti and A. albopictus. The total biting rate of these mosquito vectors was 16 bites per person per day, while the annual rate of infectious bites was 93.4. Although 2.4% for both Aedes species seems a low consumption, considering the presence of 12 odonate species at the park, it can be argued that adult odonates may play a relevant role as mosquito vector regulators, therefore impacting the spread of mosquito-borne diseases. Our study outlines the need for further research on the topic of the possible role of adult odonates for mosquito biocontrol. © 2021 Society of Chemical Industry.

Females of the red damselfly Mnesarete pudica are attracted to more ornamented males and attract rival males

Male calopterygid damselflies often exhibit colourful wings used during aggressive contests and courtship displays. Evidence suggests that male wing coloration is a secondary sexual character assessed by males and females to identify male quality. In some species, males adopt a lekking strategy, where females visit exhibition arenas and choose the best mate. Here, we addressed whether the behaviour of Mnesarete pudica males is influenced by female visitation when gathering in leks. We hypothesized that female visitation would increase male investment in courtship and fighting, while reducing patrolling flights and harassment attempts. Moreover, we tested the hypothesis that more ornamented males attract more females to the territory, following the hotshot model of lek evolution. Our results suggest that, indeed, males with more pigmented wings attract more visiting females, independently of male size. Our results also show that the number of females in a territory attracts more males and elicits male contest behaviour, reducing male harassment. We conclude that male ornament and male clustering is a good predictor of female visitation rates, suggesting that females may exert mate choice.

Loaded flight in male Ischnura elegans and its relationship to copulatory flight

Copulation in the blue-tailed damselfly (Ischnura elegans) can last several hours, during which the pair may fly together in the ‘wheel position’ with both insects flapping their wings. Previous studies have suggested that during flight in copula, the male increases its power output while the female decreases it. Consequently, the male must support some of the female’s body weight in the air. We tested the hypothesis that female body mass places a biomechanical constraint on the ability of smaller males to mate with larger females by attaching weights to male damselflies and analyzing their wing motion and force exerted using high-speed cameras. Males flying with an added load exerted extra forces equivalent to 157% of their body weight. Males flying in the mating wheel position with females whose wings were clipped bore a similar weight and were barely able to fly. To fly with an added load, males increased their wing-flapping frequency and amplitude, reaching values of mean wing tip flapping speed that were 1.9-fold higher than that in solitary flight. Our experiments indicate that although males would be able to fly briefly with the added weight of a non-responsive female, the flight performance of the pair would be severely compromised without the female contributing effort to the joint flight.

Structure and Chemical Organization in Damselfly Calopteryx haemorrhoidalis Wings: A Spatially Resolved FTIR and XRF Analysis with Synchrotron Radiation

Insects represent the majority of known animal species and exploit a variety of fascinating nanotechnological concepts. We investigated the wings of the damselfly Calopteryx haemorrhoidalis, whose males have dark pigmented wings and females have slightly pigmented wings. We used scanning electron microscopy (SEM) and nanoscale synchrotron X-ray fluorescence (XRF) microscopy analysis for characterizing the nanostructure and the elemental distribution of the wings, respectively. The spatially resolved distribution of the organic constituents was examined by synchrotron Fourier transform infrared (s-FTIR) microspectroscopy and subsequently analyzed using hierarchical cluster analysis. The chemical distribution across the wing was rather uniform with no evidence of melanin in female wings, but with a high content of melanin in male wings. Our data revealed a fiber-like structure of the hairs and confirmed the presence of voids close to its base connecting the hairs to the damselfly wings. Within these voids, all detected elements were found to be locally depleted. Structure and elemental contents varied between wing membranes, hairs and veins. The elemental distribution across the membrane was rather uniform, with higher Ca, Cu and Zn levels in the male damselfly wing membranes.

The Influence of Body Size and Agility in Displacement Capacity of Male Damselflies (Odonata, Protoneurinae)

Alternative mating tactics (AMT) occur in several animal taxa as a way to respond to physiological or environmental factors that affect reproductive success. In other words, males can exhibit different mate acquisition tactics associated with phenotypic variations. Here we present the case of Epipleoneura williamsoni, a non-territorial Neotropical damselfly in which larger males actively search for females along the margins of streams and smaller males remain perched on the vegetation and adopt a sit-and-wait strategy. Males were captured, marked and behavioral observations were conducted to take notes on the spatial displacement of individuals. We then measured male body size and agility to test whether these traits could be used to predict flight ability and thus AMT’s. Agility was measured by the ȓ2(S) coefficient, which is a surrogate of flight ability and maneuverability. The results show that body size was positively correlated with male flight ability and agility. However, male agility did not predict the adopted AMT. These results suggest that the strategies adopted by E. williamsoni males may be a reflection of body condition. This study adds evidence on how selection may act on different phenotypes within a population and how body size variations may result in different displacement capacities and flight agility.

Parasitism affects variation in a male damselfly sexual ornament

Sexually selected ornaments in animals are costly, with parasitism often affecting the degree to which they are expressed. Male Calopteryx splendens damselflies exhibit melanised ‘wing spots’. Those possessing large spots are favoured by females but also have an increased likelihood of being attacked by Calopteryx virgo, a common sympatric competitor. Melanin is used to produce the wing spots, but it is also used in immune defence against parasites that commonly infect damselflies. A total of 261 C. splendens males were collected from 26 Finnish and Latvian populations, of which half were found to be sympatric with C. virgo. It was found that males which originated from populations in which eugregarine parasites were present had smaller wing spots than individuals from parasite-free populations. Contrary to previous studies, the wing spots of C. splendens males in populations sympatric with C. virgo were not found to be smaller than those in allopatric populations. Parasite presence in C. splendens was found to be strongly associated with populations sympatric with C. virgo. The results suggest that the presence of C. virgo may increase rates of parasitism in C. splendens, and show that parasitism is an important additional factor to interspecific aggression in determining variation in the sizes of the wing spots of C. splendens males. These findings highlight a lack of understanding on the determinants of the expression of secondary sexual characters and the processes of how parasitism affects hosts.

Sexual conflict and the evolution of genitalia: male damselflies remove more sperm when mating with a heterospecific female

In Calopteryx damselflies, males remove rivals’ sperm stored by the female, thereby reducing sperm competition. This behaviour may create a sexual conflict, because females could lose the sperm stored in the spermatheca, used for long-term storage. Comparative evidence suggested antagonistic coevolution between sexes, which might prompt the evolution of narrow spermathecal ducts, or longer spermathecae, hindering sperm removal. Calopteryx haemorrhoidalis and C. splendens coexist and sometimes hybridize. Therefore, here I predicted that if females coevolve with conspecific males, heterospecific males should have an advantage when interspecific matings occur because females will show less resistance to them than to conspecific males. By hand-pairing females to males of both species, I found that in intraspecific and interspecific matings, sperm was almost completely removed from the bursa (97–100%), but only partially from the spermathecae, with more spermathecal removal in interspecific (63–71%) than intraspecific matings (14–33%). This suggests that heterospecific males are more efficient in sperm removal as predicted by a sexually-antagonistic coevolutionary scenario. Furthermore, in most cases, only the left spermatheca was emptied, suggesting that the evolution of more than one spermatheca might also be a female counter-adaptation to regain control over fertilization.

Physiological condition and wing pigmentation expression in a damselfly with seasonal polyphenism

AbstractSecondary sexual traits can be indicators of individual condition that may present seasonal polyphenism as a result of the differential costs of expression along the season. Wing spots in male damselflies of the Calopterygidae family are secondary sexual traits associated with intrasexual competition and mate choice. Hetaerina titia Drury is a calopterygid damselfly where males show red and black wing spots, contrasting with other species of the genus whose males only express a red wing spot. In the present study, we evaluate the seasonal variation of the expression of male's red and black wing spots and their allometric patterns. Additionally, we measure male condition in the form of proteins, lipids, soluble carbohydrates and glycogen in early and late seasons. Black wing spots present higher variation than red wing spots and males of the late season are more pigmented. Allometry is positive for wing red spot in the early season and for black spot in the late season. Males of the late season present a higher concentration of proteins, soluble carbohydrates and glycogen, although there is no variation in the lipid content. The results of the present study suggest that, in H. titia males, black pigmentation replaces the function of the red pigmentation to signal condition. Both traits, however, may be heavily affected by environmental situations (e.g. food availability).

Role of visual and non-visual cues in damselfly mate recognition

In many species of damselflies, sexual conflict in the form of male harassment is thought to explain the widespread existence of female-limited color polymorphisms. With a few exceptions, the majority of investigations into these mating systems have assumed that male damselflies primarily use visual cues to detect and recognize their mates. Recently, laboratory studies have demonstrated that damselflies orient to olfactory signals from prey and that males orient to chemical cues from conspecific females. However, to date there are no field experiments which explicitly test the role that chemical cues play in sex recognition. Here we used a field experiment on Enallagma civile damselflies to test if free flying males detect and recognize females in the absence of visual cues through the use of non-visual signals. In the absence of visual cues males did not exhibit positive responses toward female conspecifics, whereas when both visual and non-visual cues were present, males readily detected females and oft...

The price of looking sexy: visual ecology of a three‐level predator–prey system

Summary Colour signals and colour vision play a pivotal role in intraspecific communication and predator–prey interactions. However, the costs of expressing conspicuous sexual signals at multiple trophic levels have been largely overlooked. Sexual signals can also experience character displacement in sympatric populations of closely related species, leading to potential changes in conspicuousness. We here investigate a bird–damselfly–fruit fly predator–prey system, where two closely related damselfly species have conspicuous, sexually selected wing coloration. The damselflies can occur in sympatry and allopatry, and reproductive character displacement in the coloration size has been previously reported. We quantify the damselfly wing reflectance from replicated sympatric and allopatric populations, and use receptor noise models to investigate the visual discriminability of the wing coloration for the bird, damselfly and fly vision systems, against natural backgrounds. We perform electroretinograms to study damselfly eye sensitivity. We also estimate damselfly predation risk in natural populations. We find that the chromatic component of wing coloration makes males highly discriminable to the predator, but not to the prey. However, female wing coloration is predominantly cryptic for the predator and prey, and interestingly, also for male damselflies. A female being cryptic to conspecifics likely reduces male harassment. The estimates of predation risk partially support the discriminability results. We also show that there is no difference in colour vision sensitivity between the two damselfly species and sexes, and no difference in wing coloration or its discriminability between sympatric and allopatric populations. Our results suggest that sexually selected traits can be antagonistically selected by predators and prey and that this antagonistic selection can be sex‐dependent: males are paying a large cost in terms of conspicuousness, while females remain mostly cryptic. Our study thus emphasizes the need for investigating visual communication at multitrophic levels since the degree of colour discriminability can differ between predators, prey and the focal species. A lay summary is available for this article.

Variations of space use in males of the banded demoiselle (Calopteryx splendens, Zygoptera, Odonata): Alternative tactics or an age-dependent trend?

Two spatial tactics are usually distinguished in males of Calopteryx damselflies: territorial and nonterritorial. These tactics are believed to underlie two alternative condition-dependent reproductive tactics in these insects, and territorial males are believed to copulate more often. With age, males become weaker, turn nonterritorial, and only occasionally manage to copulate. However, the details of space use by damselflies are poorly known, which hinders the interpretation of the existing empirical data. We describe the space use by individually marked males of the banded demoiselle C. splendens studied during three field seasons in Vladimir Province, Russia. Each male on each day of observations was characterized as either territorial or non-territorial, and the sites of encounter were mapped. The probability of being territorial declined with the male’s age. The spatial tactics (territorial vs. non-territorial) on a given day strongly influenced the tactics used on the following day. We identified the territorial and non-territorial phases in the life of a male damselfly, which occurred consecutively and had a roughly similar duration. During the territorial phase, the male occupied a certain territory and tried to hold it as long as possible. The male abandoned its territory in two cases: (1) when it was driven onto a different territory as the result of competition with other males, or (2) when it was exhausted and became non-territorial. Thus, the space use by the male changed predictably during its life. Therefore, direct comparison of morphological or other characteristics in territorial vs. non-territorial males, frequently made in the literature, makes little sense. Further progress in studying the so-called “alternative reproductive tactics” in damselflies may be more successfully achieved by comparing individual life trajectories of different males (e.g. duration of territorial and non-territorial periods, the number of consecutively occupied territories, etc.). We performed correlation analysis and found that the above parameters did not depend on the wing and abdomen length of the males.

Rubyspot Territorial Damselflies Behave as “Nasty Neighbors”

Two mutually-excluding hypotheses explain the intensity of aggression between neighbors and their non-neighbors in territorial animals. On one hand, the “dear enemy” hypothesis predicts that territorial animals should be more tolerant towards their neighbors than towards non-neighbors. Conversely, the “nasty neighbor” hypothesis predicts increased aggression towards neighbors than non-neighbors. These different situations depend on who is more likely to be a real competitor, either a neighbor or a non-neighbor, and the intensity of resource competition. Male damselfly of Hetaerina vulnerata defend riverine, mating territories that is the main way to have access to females. These territories are not fixed and so males continuously defend them especially against neighbors. Given this, we tested whether the nasty neighbor principle operates in this species. We monitored a population during an entire mating season, and recorded duration of aggressive behaviors. As a key prediction of the nasty neighbor hypothesis, we expected that such behaviors last for longer when encountering a conspecific neighbor than a conspecific non-neighbor. We also predicted that such duration should be date-dependent as territorial competition could increase in the middle of the season when male density is at its highest. Our results corroborated that aggressive behaviors lasted longer against a neighbor male than a non-neighbor male but there was not effect of date. Thus, neighbors may pose a greater risk and may be a strong selective force than non-neighbors in terms of resource competition. Since neighbor males are continuously trying to widen their territory boundaries (a situation that has been also found in other study systems), an owner male may not even need to recognize his neighbors to fight back.

Functional morphology of the male caudal appendages of the damselfly Ischnura elegans (Zygoptera: Coenagrionidae)

Odonata are usually regarded as one of the most ancient extant lineages of winged insects. Their copulatory apparatus and mating behavior are unique among insects. Male damselflies use their caudal appendages to clasp the female's prothorax during both copulation and egg-laying and have a secondary copulatory apparatus for sperm transfer. Knowledge of the functional morphology of the male caudal appendages is the basis for understanding the evolution of these structures in Odonata and respective organs in other insects. However, it is still not exactly known how the zygopteran claspers work. In this study, we applied micro-computed tomography and a variety of microscopy techniques to examine the morphology, surface microstructure, cuticle material composition and muscle topography of the male caudal appendages of Ischnura elegans. The results indicate that the closing of the paraproctal claspers is mainly passive. This indirect closing mechanism is very likely supported by high proportions of the elastic protein resilin present in the cuticle of the paraproctal bases. In addition, the prothoracic morphology of the female plays an important role in the indirect closing of the male claspers. Our data indicate that both structures – the male claspers and the female prothoracic hump – function together like a snap-fastener.

Sympatric shift in a male sexual ornament in the damselfly Calopteryx splendens

Character displacement is a process by which interactions between two species that exhibit similar traits, results in geographical patterns of trait divergence in one or both species. These traits evolve to reduce costs of interspecific interactions in sympatry and thus differ from their condition in allopatry. In male damselflies Calopteryx splendens, large wing spots are sexually selected. However, in sympatric populations with Calopteryx virgo, wing spot size decreases as C.virgo abundance increases. The stability of this pattern is unclear, because previous studies have focused on sympatric populations with potentially fluctuating relative abundances. We studied the wing spot sizes of C.splendens in both sympatric and allopatric populations. Our data show that male C.splendens' wing spots are larger in allopatry than in sympatry with C.virgo. We suggest that both interspecific aggression and avoidance of interspecific reproductive interactions may result in this pattern, although their relative importance remains unclear.

Predation selects for increased immune function in male damselflies, Calopteryx splendens.

Predation selects for numerous traits in many animal species, with sick or parasitized prey often being at high risk. When challenged by parasites and pathogens, prey with poor immune functions are thus likely to be at a selective disadvantage. We tested the hypothesis that predation by birds selects for increased immune function in a wild population of male damselflies Calopteryx splendens, while controlling for a trait known to be under selection by bird predation, dark wing-spots. We found that selection on both immune function and wing-spot size was significantly positive, and that selection on either trait was independent of selection on the other. We found no evidence of nonlinear quadratic or correlational selection. In contrast to previous studies, we found no phenotypic correlation between immune function and wing-spot size. There was also no difference in immune response between territorial and non-territorial males. Our study suggests that predation may be an important agent of selection on the immune systems of prey, and because the selection we detected was directional, has the potential to cause phenotypic change in populations.

Activation of the immune system promotes insect dispersal in the wild

Dispersal has important ecological and evolutionary consequences but is a poorly understood behaviour. We experimentally tested whether activation of the immune system affects dispersal in male damselflies, Calopteryx virgo, from three natural populations. We show that males that contained an experimentally inserted artificial pathogen, a nylon monofilament implant, had higher dispersal rates and flew further than control males, but not further than sham manipulated males. Our data suggest that dispersal may reduce the risk of further infections if immune system activation indicates high parasite infection risk in the present habitat. We, thus, suggest that parasites may play an important role in the evolution of host dispersal.

Effects of extreme variation in female morph frequencies on the mating behaviour of male damselflies

Female-limited polymorphism is often attributed to selection to avoid excessive male mating attempts. It is encountered in various taxonomic groups, but is particularly common in damselflies, where one female morph (andromorph) typically resembles the conspecific male in colour pattern, while the other(s) (gynomorph(s)) do not. Two sets of theories have been proposed to explain the phenomenon in damselflies, which can be classified as the learned mate recognition (LMR) and male mimicry (MM) hypotheses. To test predictions of these hypotheses, we evaluated the rate of male sexual response towards female morphs and conspecific males in the damselfly Nehalennia irene. The LMR hypothesis predicts that males should respond sexually to andromorphs at greater rates in populations containing a higher relative frequency of andromorphs. The MM hypothesis predicts that males respond more often sexually to both andromorphs and males as the ratio of andromorphs to males increases. While LMR predicts that the rate of mating attempts towards gynomorphs should vary, the MM predicts that it should be relatively fixed. On experimentally presenting live specimens to focal males in five different populations with extreme variation in female morph frequencies, we observed that as the andromorph frequency and ratio of andromorphs to males increased, the proportion of male mating attempts increased on both andromorphs and males, whereas it decreased on gynomorphs. While the simplest form of the MM hypothesis is rejected, the results support specific predictions of both hypotheses and suggest that future studies should not treat these hypotheses as mutually exclusive.

Influence of alternative mating tactics on predation risk in the damselfly Calopteryx virgo

Alternative mating tactics are a widespread feature in insects. A typical form of alternative mating behaviour is being a sneaker in the vicinity of a territorial male. Such nonterritorial males have lower mating success, but they may benefit from lower energetic costs and decreased predation risk. In this study, we examined whether nonterritorial male damselflies Calopteryx virgo (L., 1758) are subject to lower predation risk than territorial males. To distinguish predation from other sources of mortality, we used models. The experiment consisted of dried male damselflies settled into the typical perching positions of territorial and nonterritorial males. Also the spatiotemporal patterns of predation risk were studied. The survival of nonterritorial male models was consistently higher than that of territorial male models, which can be attributed to different predation risk. Survival of the models was lower in the presence of avian predators and in large populations. Survival rates were affected by habitat type but did not change during the season. We conclude that nonterritorial male damselflies are less vulnerable to predation and that there may be a trade-off which could potentially make the fitness of sneakers equal to that of territorial males.