Natural Selection on Adults Has Trait-Dependent Consequences for Juvenile Evolution in Dragonflies.

AbstractAlthough natural selection often fluctuates across ontogeny, it remains unclear what conditions enable selection in one life-cycle stage to shape evolution in others. Organisms that undergo metamorphosis are useful for addressing this topic because their highly specialized life-cycle stages cannot always evolve independently despite their dramatic life-history transition. Using a comparative study of dragonflies, we examined three conditions that are hypothesized to allow selection in one stage to affect evolution in others. First, we tested whether lineages with less dramatic metamorphosis (e.g., hemimetabolous insects) lack the capacity for stage-specific evolution. Rejecting this hypothesis, we found that larval body shape evolves independently from selection on adult shape. Next, we evaluated whether stage-specific evolution is limited for homologous and/or coadapted structures. Indeed, we found that selection for larger wings is associated with the evolution of coadapted larval sheaths that store developing wing tissue. Finally, we assessed whether stage-specific evolution is restricted for traits linked to a single biochemical pathway. Supporting this hypothesis, we found that species with more wing melanization in the adult stage have evolved weaker melanin immune defenses in the larval stage. Thus, our results collectively show that natural selection in one stage imposes trait-dependent constraints on evolution in others.

Atrazine Exposure Influences Immunity in the Blue Dasher Dragonfly, Pachydiplax longipennis (Odonata: Libellulidae).

Agricultural runoff containing herbicide is known to have adverse effects on freshwater organisms. Aquatic insects are particularly susceptible, and herbicide runoff has the potential to affect immunity in this group. Here we examined the effect of ecologically relevant levels of atrazine, an herbicide commonly used in the United States, on immune function in larvae of the blue dasher dragonfly (Odonata: Libelluludae, Pachydiplax longipennis Burmeister 1839) during a long-term exposure at ecologically relevant concentrations. Larvae were exposed to concentrations of 0, 1, 5, and 10 ppb atrazine for 3 or 6 wk. Hemocyte counts, hemolymph phenyloxidase (PO) activity, cuticular PO, and gut PO were measured at the end of each trial period as indicators of immune system strength. Atrazine concentration had a significant effect on hemocyte counts after controlling for larval size. There was a significant interaction between time and concentration for hemolymph PO, cuticular PO, and a marginal interaction for gut PO. The effect of atrazine on the measured immune parameters was often nonmonotonic, with larger effects observed at intermediate concentrations. Therefore, atrazine affects both hemocyte numbers and PO activity over time in P. longipennis, and the changed immune function demonstrated in this study is likely to modify susceptibility to pathogens, alter wound healing, and may decrease available energy for growth and metamorphosis.

Immune deployment increases larval vulnerability to predators and inhibits adult life-history traits in a dragonfly.

While deploying immune defences early in ontogeny can trade-off with the production and maintenance of other important traits across the entire life cycle, it remains largely unexplored how features of the environment shape the magnitude or presence of these lifetime costs. Greater predation risk during the juvenile stage may particularly influence such costs by (1) magnifying the survival costs that arise from any handicap of juvenile avoidance traits and/or (2) intensifying allocation trade-offs with important adult traits. Here, we tested for predator-dependent costs of immune deployment within and across life stages using the dragonfly, Pachydiplax longipennis. We first examined how larval immune deployment affected two traits associated with larval vulnerability to predators: escape distance and foraging under predation risk. Larvae that were induced to mount an immune response had shorter escape distances but lower foraging activity in the presence of predator cues. We also induced immune responses in larvae and reared them through emergence in mesocosms that differed in the presence of large predatory dragonfly larvae (Aeshnidae spp.). Immune-challenged larvae had later emergence overall and lower survival in pools with predators. Immune-challenged males were also smaller at emergence and developed less sexually selected melanin wing coloration, but these effects were independent of predator treatment. Overall, these results highlight how mounting an immune defence early in ontogeny can have substantial ecological and physiological costs that manifest both within and across life stages.

Beneficial effects of a heat wave: higher growth and immune components driven by a higher food intake.

While heat waves will become more frequent and intense under global warming, the ability of species to deal with extreme weather events is poorly understood. We investigated how a heat wave influenced growth rate and investment in two immune components (phenoloxidase activity and melanin content) in larvae of two damselfly species, Ischnura elegans and Enallagma cyathigerum Late instar larvae were kept at 18°C (i.e. their average natural water temperature) or under a simulated long heat wave at 30°C. To explain the heat wave effects, we quantified traits related to energy uptake (food intake and growth efficiency), energy expenditure (metabolic rate measured as activity of the electron transport system, ETS) and investment in energy storage (fat content). The two species differed in life strategy, with I. elegans having a higher growth rate, growth efficiency, ETS activity and fat content. In line with its preference for cooler water bodies, the heat wave was only lethal for E. cyathigerum However, both species benefited from the heat wave by increasing growth rate, which can be explained by the higher increase in food intake than metabolic rate. This may also have contributed to the increased investment in energy storage and immune components under the heat wave. This mediatory role of food intake indicates the critical role of food availability and behaviour in shaping the impact of heat waves. Our results highlight the importance of including behavioural and physiological variables to unravel and predict the impact of extreme climate events on organisms.

Predation risk increases immune response in a larval dragonfly (Leucorrhinia intacta).

Predators often negatively affect prey performance through indirect, non-consumptive effects. We investigated the potential relationship between predator-induced stress and prey immune response. To test this, we administered a synthetic immune challenge into dragonfly larvae (Leucorrhinia intacta) and assessed a key immune response (level of encapsulation) in the presence and absence of a caged predator (Anax junius) at two temperatures (22 degrees C and 26 degrees C). We hypothesized that immune response would be lowered when predators were present due to lowered allocation of resources to immune function and leading to reduced encapsulation of the synthetic immune challenge. Contrary to our expectations, larvae exposed to caged predators had encapsulated monofilaments significantly more than larvae not exposed to caged predators. Levels of encapsulation did not differ across temperatures, nor interact with predator exposure. Our results suggest that the previously observed increase in mortality of L. intacta exposed to caged predators is not driven by immune suppression. In situations of increased predation risk, the exposure to predator cues may induce higher levels of melanin production, which could lead to physiological damage and high energetic costs. However, the costs and risks of increased allocations to immune responses and interactions with predation stress remain unknown.

Rapid evolution of larval life history, adult immune function and flight muscles in a poleward-moving damselfly.

Although a growing number of studies have documented the evolution of adult dispersal-related traits at the range edge of poleward-expanding species, we know little about evolutionary changes in immune function or traits expressed by nondispersing larvae. We investigated differentiation in larval (growth and development) and adult traits (immune function and flight-related traits) between replicated core and edge populations of the poleward-moving damselfly Coenagrion scitulum. These traits were measured on individuals reared in a common garden experiment at two different food levels, as allocation trade-offs may be easier to detect under energy shortage. Edge individuals had a faster larval life history (growth and development rates), a higher adult immune function and a nearly significant higher relative flight muscle mass. Most of the differentiation between core and edge populations remained and edge populations had a higher relative flight muscle mass when corrected for latitude-specific thermal regimes, and hence could likely be attributed to the range expansion process per se. We here for the first time document a higher immune function in individuals at the expansion front of a poleward-expanding species and documented the rarely investigated evolution of faster life histories during range expansion. The rapid multivariate evolution in these ecological relevant traits between edge and core populations is expected to translate into changed ecological interactions and therefore has the potential to generate novel eco-evolutionary dynamics at the expansion front.

Immune- and wound-dependent differential gene expression in an ancient insect.

Two of the main functions of the immune system are to control infections and to contribute to wound closure. Here we present the results of an RNAseq study of immune- and wound-response gene expression in the damselfly Coenagrion puella, a representative of the odonates, the oldest taxon of winged insects. De novo assembly of RNAseq data revealed a rich repertoire of canonical immune pathways, as known from model insects, including recognition, transduction and effector gene expression. A shared set of immune and wound repair genes were differentially expressed in both wounded and immune-challenged larvae. Moreover 3-fold more immune genes were induced only in the immune-challenged treatment. This is consistent with the notion that the immune-system reads a balance of signals related to wounding and infection and that the response is tailored accordingly.

Support for the immunocompetence handicap hypothesis in the wild: hormonal manipulation decreases survival in sick damselflies.

The immunocompetence handicap hypothesis (ICHH) states that hormones enhance sexual trait expression but impair immunity. Previous tests of the ICHH have been hampered by experimental design problems. Here, we report on an experimental test of the ICHH that includes manipulations of both hormones and infections in males of the territorial damselfly, Hetaerina americana, with accurate survival measurements. We conducted a fully factorial experiment subjecting each individual to one of three topical treatments: methoprene (a juvenile hormone analog), acetone, or control, and one of three injection treatments: bacteria, PBS, or control. We measured survival of manipulated males in both the wild and in captivity. As predicted, survival was most heavily impaired in methoprene-bacteria males than in the other groups in the wild, and no survival differences emerged in captive animals. This result confirms that survival is one cost an animal pays for increased hormonal levels. This corroborates theoretical predictions of the ICHH.