Inducible Morphological Defences Research Articles

Uncovering the chemistry behind inducible morphological defences in the crustacean Daphniamagna via micro-Raman spectroscopy

The widespread distribution of Crustacea across every aquatic ecological niche on Earth is enabled due to their exoskeleton's versatile properties. Especially mineralization of the exoskeleton provides protection against diverse environmental threats. Thereby, the exoskeleton of some entomostracans is extremely phenotypically plastic, especially in response to predators. For instance, the freshwater zooplankton Daphnia forms conspicuous inducible morphological defenses, such as helmets, and can increase the stability of its exoskeleton, which renders them less vulnerable to predation. In this study, we reveal for the first time the chemical composition of the exoskeleton of Daphniamagna, using Raman spectroscopy, to be composed of α-chitin and proteins with embedded amorphous calcium carbonate (ACC). Furthermore, we reveal the exoskeleton's chemical changes associated with inducible defense mechanisms in the form of more substantial mineralization, which is probably correlated with enhanced carapace stability. We, therefore, highlight the importance of calcium-biominerals for inducible morphological defenses in Daphnia.

Inducible morphological defense in Daphnia pulex: food quantity effects revised

In aquatic systems, organisms largely rely on chemical cues to perceive information about the presence of predators or prey. Daphnia recognize the presence of the predatory larvae of Chaoborus via a chemical cue, emitted by the larvae, a so-called kairomone. Upon recognition, neckteeth, an alteration of the carapace, are induced in Daphnia that reduce predation rates of Chaoborus. Neckteeth induction was often reported to entail costs. In a previous study, food quantity affected the level of neckteeth induction, with stronger neckteeth induction at low food concentrations and weak induction at high food concentrations. However, reducing neckteeth induction at high food quantities seems to be maladaptive and not in accordance with the concept that inducible defenses are associated with costs.Here, we hypothesized that weaker neckteeth induction at high food concentrations is caused by increased bacterial degradation of the kairomone. More specifically, we assume that higher algal food concentration is associated with higher bacterial abundances, which degrade the kairomone during the experiment. We tested our hypothesis by treating food algae with antibiotics before providing them as food to Daphnia. Antibiotics reduced bacterial abundances at high and low food concentrations. Reduced bacterial abundances at high food concentrations led to the same level of neckteeth induction as at low food concentrations. A linear regression revealed a significant correlation of neckteeth induction to bacterial abundances. We therefore conclude that differences in neckteeth induction at different food concentrations are not caused by the food quantity effects but by differences in bacterial degradation of the kairomone.

Size‐selective predation accounts for intra‐ and inter‐specific variation of inducible morphological defense of Daphnia

AbstractInducible defense is adaptive when prey organisms cope with fluctuations of predation risk, and there exists ample variations of inducible defense both within a species and between species even in a single habitat. Daphnia, a freshwater zooplankton genus, shows remarkable inducible morphological defense when exposed to predators. This study explored the intra‐ and inter‐specific variations of inducible defense of Daphnia and tested whether the variations can be explained by the size‐selective nature of predation. In Lake Fukami‐ike, Japan, Daphnia ambigua and Daphnia pulex coexist with planktivorous fish and Chaoborus larvae that have different preferences for prey size. We experimentally examined the ontogenetic change of inducible defense for ten clones of each of the two Daphnia species against the two different predators. The degree of defense expression evaluated based on the induction of multiple morphological traits showed remarkable differences between Daphnia species, instars and clones, and the intra‐ and inter‐specific variations were attributed to size‐selective predation from the different predators. The inducible defense of Daphnia showed the adaptive variations depending on the predation pressure that resulted from the size‐selective predation by the coexisting predators.

Predation risk induces age- and sex-specific morphological plastic responses in the fathead minnow Pimephales promelas

Although comprehending the significance of phenotypic plasticity for evolution is of major interest in biology, the pre-requirement for that, the understanding of variance in plasticity, is still in its infancy. Most researchers assess plastic traits at single developmental stages and pool results between sexes. Here, we study variation among sexes and developmental stages in inducible morphological defences, a well-known instance of plasticity. We raised fathead minnows, Pimephales promelas, under different levels of background predation risk (conspecific alarm cues or distilled water) in a split-clutch design and studied morphology in both juveniles and adults. In accordance with the theory that plasticity varies across ontogeny and sexes, geometric morphometry analyses revealed significant shape differences between treatments that varied across developmental stages and sexes. Alarm cue-exposed juveniles and adult males developed deeper heads, deeper bodies, longer dorsal fin bases, shorter caudal peduncles and shorter caudal fins. Adult alarm cue-exposed males additionally developed a larger relative eye size. These responses represent putative adaptive plasticity as they are linked to reduced predation risk. Perhaps most surprisingly, we found no evidence for inducible morphological defences in females. Understanding whether similar variation occurs in other taxa and their environments is crucial for modelling evolution.

Defence versus defence: Are crucian carp trading off immune function against predator-induced morphology?

Numerous species adopt inducible defence strategies; that is, they have phenotypically plastic traits that decrease the risk of capture and consumption by potential predators. The benefits of expressing alternative phenotypes in high- vs. low-risk environments are well documented. However, inducible anti-predator traits are also expected to incur costs, as they are not expressed when predators are absent, yet empirical evidence of such costs remains scarce. Virtually, all animals in nature are simultaneously under strong selection to evade both capture by predators and infection by parasites or pathogens and, hence, display a diverse arsenal of defences to combat these threats, raising the possibility of trade-offs between defences. A classic example of a predator-induced morphological defence is the deep-bodied shape of crucian carp that reduces risk of predation from gape-size-limited predators. The goal of this study was to examine whether predator exposure affects also immune function in crucian carp, and whether the degree of expressed morphological defence is traded off against immune function in individuals. Following exposure to manipulations of perceived risk (predator presence/absence) in a long-term experiment (8months), key aspects of innate immune function and individual differences in the expression of inducible morphological defence were quantified. Predator-exposed individuals showed lower haptoglobin levels and complement activity, but higher natural antibody titres than fish from predator-free conditions. When experimentally challenged with a mimicked bacterial infection (LPS injection), fish reared in the presence of a natural predator showed a weaker immune response. Moreover, among predator-exposed individuals, the magnitude of morphological defence expression correlated with both baseline immune function and the ability to mount an immune response. However, these relationships were not consistently supportive of a general trade-off among defences. Our results suggest that fish exposed to predators on average reduce investment in immune function, and, further, the observed relationships among defences in predator-exposed individuals can best be explained from individual fitness and pace-of-life perspectives.

Biomechanical properties of predator-induced body armour in the freshwater crustacean Daphnia

The freshwater crustacean Daphnia is known for its ability to develop inducible morphological defences that thwart predators. These defences are developed only in the presence of predators and are realized as morphological shape alterations e.g. ‘neckteeth’ in D. pulex and ‘crests’ in D. longicephala. Both are discussed to hamper capture, handling or consumption by interfering with the predator’s prey capture devices. Additionally, D. pulex and some other daphniids were found to armour-up and develop structural alterations resulting in increased carapace stiffness. We used scanning transmission electron microscopy (STEM) and confocal laser scanning microscopy (CLSM) to identify predator-induced structural and shape alterations. We found species specific structural changes accompanying the known shape alterations. The cuticle becomes highly laminated (i.e. an increased number of layers) in both species during predator exposure. Using nano- and micro-indentation as well as finite element analysis (FEA) we determined both: the structure’s and shape’s contribution to the carapace’s mechanical resistance. From our results we conclude that only structural alterations are responsible for increased carapace stiffness, whereas shape alterations appear to pose handling difficulties during prey capture. Therefore, these defences act independently at different stages during predation.

Evolution of a predator-induced, nonlinear reaction norm.

Inducible, anti-predator traits are a classic example of phenotypic plasticity. Their evolutionary dynamics depend on their genetic basis, the historical pattern of predation risk that populations have experienced and current selection gradients. When populations experience predators with contrasting hunting strategies and size preferences, theory suggests contrasting micro-evolutionary responses to selection. Daphnia pulex is an ideal species to explore the micro-evolutionary response of anti-predator traits because they face heterogeneous predation regimes, sometimes experiencing only invertebrate midge predators and other times experiencing vertebrate fish and invertebrate midge predators. We explored plausible patterns of adaptive evolution of a predator-induced morphological reaction norm. We combined estimates of selection gradients that characterize the various habitats that D. pulex experiences with detail on the quantitative genetic architecture of inducible morphological defences. Our data reveal a fine scale description of daphnid defensive reaction norms, and a strong covariance between the sensitivity to cues and the maximum response to cues. By analysing the response of the reaction norm to plausible, predator-specific selection gradients, we show how in the context of this covariance, micro-evolution may be more uniform than predicted from size-selective predation theory. Our results show how covariance between the sensitivity to cues and the maximum response to cues for morphological defence can shape the evolutionary trajectory of predator-induced defences in D. pulex.

Inducible Defenses with a "Twist": Daphnia barbata Abandons Bilateral Symmetry in Response to an Ancient Predator

Predation is one of the most important drivers of natural selection. In consequence a huge variety of anti-predator defenses have evolved in prey species. Under unpredictable and temporally variable predation pressure, the evolution of phenotypically plastic defensive traits is favored. These “inducible defenses”, range from changes in behavior, life history, physiology to morphology and can be found in almost all taxa from bacteria to vertebrates. An important group of model organisms in ecological, evolutionary and environmental research, water fleas of the genus Daphnia (Crustacea: Cladocera), are well known for their ability to respond to predators with an enormous variety of inducible morphological defenses. Here we report on the “twist”, a body torsion, as a so far unrecognized inducible morphological defense in Daphnia, expressed by Daphnia barbata exposed to the predatory tadpole shrimp Triops cancriformis. This defense is realized by a twisted carapace with the helmet and the tail spine deviating from the body axis into opposing directions, resulting in a complete abolishment of bilateral symmetry. The twisted morphotype should considerably interfere with the feeding apparatus of the predator, contributing to the effectiveness of the array of defensive traits in D. barbata. As such this study does not only describe a completely novel inducible defense in the genus Daphnia but also presents the first report of a free living Bilateria to flexibly respond to predation risk by abandoning bilateral symmetry.

Phenotypic plasticity in three Daphnia genotypes in response to predator kairomone: evidence for an involvement of chitin deacetylases.

The genetic background of inducible morphological defences in Daphnia is still largely unknown. Dissolved infochemicals from the aquatic larvae of the phantom midge Chaoborus induce so-called 'neck-teeth' in the first three post-embryonic stages of Daphnia pulex This defence has become a textbook example of inducible defences. In a target gene approach, by using three Daphnia genotypes which show a gradient of neck-teeth induction in response to equal amounts of kairomone, we report a high correlation of neck-teeth induction in Daphnia pulex and relative gene expression of two chitin deacetylases. Further, previous studies suggested genes from both the juvenoid and the insulin hormone signalling pathways as well as several morphogenetic genes downstream to be responsible for neck-teeth induction in D. pulex However, these data were not supported by our study. None of the three D. pulex clones showed an upregulation of these previously proposed candidate genes as a response to predator kairomone, which is interpreted as the result of refined methods used for both RNA sampling and kairomone enrichment yielding unambiguous results compared with earlier studies. The assessment of a clonal gradient of Daphnia in the presence and absence of infochemicals provides a promising approach to identify further genes involved in the induction of morphological defences by correlating gene expression and morphology.

Rapid evolution of constitutive and inducible defenses against an invasive predator.

Invasive alien predators can impose strong selection on native prey populations and induce rapid evolutionary change in the invaded communities. However, studies on evolutionary responses to invasive predators are often complicated by the lack of replicate populations differing in coexistence time with the predator, which would allow the determination of how prey traits change during the invasion. The red swamp crayfish Procambarus clarkii has invaded many freshwater areas worldwide, with negative impacts for native fauna. Here, we examined how coexistence time shapes antipredator responses of the Iberian waterfrog (Pelophylax perezi) to the invasive crayfish by raising tadpoles from five populations differing in historical exposure to P. clarkii (30 years, 20 years, or no coexistence). Tadpoles from non-invaded populations responded to the presence of P. clarkii with behavioral plasticity (reduced activity), whereas long-term invaded populations showed canalized antipredator behavior (constant low activity level). Tadpoles from one of the long-term invaded populations responded to the crayfish with inducible morphological defenses (deeper tails), reflecting the use of both constitutive and inducible antipredator defenses against the exotic predator by this population. Our results suggest that, while naive P. perezi populations responded behaviorally to P. clarkii, the strong predation pressure imposed by the crayfish has induced the evolution of qualitatively different antipredator defenses in populations with longer coexistence time. These responses suggest that strong selection by invasive predators may drive rapid evolutionary change in invaded communities. Examining responses of prey species to biological invasions using multiple populations will help us better forecast the impact of invasive predators in natural communities.

Increasing conspecific density weakens the ability of intermediate predators to develop induced morphological defences to top predators

SummaryIntraguild predation is common in nature, but it is unclear how species that both compete and eat each other can persist together. One possibility is that intermediate predators possess inducible morphological defences that protect them from top predators while not compromising their ability to compete with top predators.The ability of intermediate predators to develop morphological defences may be compromised in environments with a high density of conspecifics because of reduced resource availability and predation risk due to the saturating functional response of top predators. Furthermore, since morphological defences take time to develop, the type and extent of morphological defences may vary during development.We conducted an experiment to measure the phenotypic responses of an intermediate predator (larvae of the salamanderAmbystoma opacum) to the presence of a caged top predator (larvae of the dragonflyAnaxspp.) throughout ontological development in environments that differed in the density of conspecifics present. We also assessed how intermediate predators, reared in the different environments, differed in their vulnerability to top predators and ability to deplete their food resources.We found thatAnaxinduced morphological defences inA. opacum, but the extent of morphological change declined with the density of conspecifics. Moreover, some morphological traits disappeared, while others appeared just prior toA. opacummetamorphosis. The change inA. opacumphenotype in response toAnaxmadeA. opacumless vulnerable to predation byAnaxbut had no significant effect on the foraging ability ofA. opacum.Our study demonstrates that top predators can induce phenotypes in intermediate predators that reduce their vulnerability to top predators while not compromising their ability to feed on a common prey. An increase in intermediate predator density, however, could diminish the ability of intermediate predators to develop the full suite of morphological defences. The inability to develop the full suite of morphological defences may reduce the probability of persistence with top predators.

Uncovering Ultrastructural Defences in Daphnia magna – An Interdisciplinary Approach to Assess the Predator-Induced Fortification of the Carapace

The development of structural defences, such as the fortification of shells or exoskeletons, is a widespread strategy to reduce predator attack efficiency. In unpredictable environments these defences may be more pronounced in the presence of a predator. The cladoceran Daphnia magna (Crustacea: Branchiopoda: Cladocera) has been shown to develop a bulky morphotype as an effective inducible morphological defence against the predatory tadpole shrimp Triopscancriformis (Crustacea: Branchiopoda: Notostraca). Mediated by kairomones, the daphnids express an increased body length, width and an elongated tail spine. Here we examined whether these large scale morphological defences are accompanied by additional ultrastructural defences, i.e. a fortification of the exoskeleton. We employed atomic force microscopy (AFM) based nanoindentation experiments to assess the cuticle hardness along with tapping mode AFM imaging to visualise the surface morphology for predator exposed and non-predator exposed daphnids. We used semi-thin sections of the carapace to measure the cuticle thickness, and finally, we used fluorescence microscopy to analyse the diameter of the pillars connecting the two carapace layers. We found that D . magna indeed expresses ultrastructural defences against Triops predation. The cuticle in predator exposed individuals is approximately five times harder and two times thicker than in control daphnids. Moreover, the pillar diameter is significantly increased in predator exposed daphnids. These predator-cue induced changes in the carapace architecture should provide effective protection against being crushed by the predator’s mouthparts and may add to the protective effect of bulkiness. This study highlights the potential of interdisciplinary studies to uncover new and relevant aspects even in extensively studied fields of research.

Intraspecific trait cospecialization of constitutive and inducible morphological defences in a marine snail from habitats with different predation risk

1. Studies examining the integration of constitutive and inducible aspects of multivariate defensive phenotypes are rare. 2. I asked whether marine snails (Nucella lamellosa) from habitats with and without abundant predatory crabs differed in constitutive and inducible aspects of defensive shell morphology. 3. I examined multivariate shell shape development of snails from each habitat in the presence and absence of waterborne cues from feeding crabs (Cancer productus). I also examined the influence of constitutive and inducible shell morphology on resistance to crushing. 4. Regardless of the presence of crabs, snails from high-risk (HR) habitats developed rotund, short-spired shells, while snails from low-risk habitats developed elongate shells, tall-spired shells, indicating among-habitat divergence in constitutive shell shape. Moreover, allometry analyses indicated that constitutive developmental patterns underlying this variation also differed between habitats. However, snails from HR habitats showed greater plasticity for apertural lip thickness and apertural area in the presence of crab cues, indicating among-habitat variation in defence inducibility. 5. Both shell shape and apertural lip thickness contributed to shell strength suggesting that constitutive shell shape development and inducible lip thickening have evolved jointly to form an effective defence in habitats where predation risk is high.

Growing large and bulky in the presence of the enemy: Daphnia magna gradually switches the mode of inducible morphological defences

Summary 1. Phenotypic plasticity in defensive traits has been proven to be effective in ecosystems with frequently changing predator regimes. However, if a single dominant predator exerts predation pressure on each ontogenetic stage, prey should adapt by developing defensive traits for each life stage within a cost-benefit framework. This may require a change of defensive mechanisms between juvenile and adult life stages. 2. In this study, we examined the morphological defences of the cladoceran Daphnia magna Strauss induced by the tadpole shrimp Triops cancriformis Bosc. We tested for the induction of morphological defences by directly exposing daphnids to the predator and conducted a life span experiment to determine if the expression of the induced morphological defences varies throughout the life span of D. magna. In addition, we studied the adaptive value, i.e. the effectiveness, of the Triops-induced morphological defences in D. magna by conducting predation trials. 3. We found that, in D. magna, the expression of an array of inducible morphological defences, which act synergistically to provide effective protection, changes during the daphnids lifetime in response to the tadpole shrimp T. cancriformis. This gradual switch in the protective function of single traits between juvenile and adult stages represents a novel functionality and complexity of inducible defences. Both direct contact with the predator and chemical cues (kairomones) released by T. cancriformis induce an increased body length, body width and an elongation of the tail spine in D. magna. This study is the first to show that kairomones released by a predator can induce ‘bulkiness’ as a defensive mechanism in Daphnia. Finally, we demonstrate the effectiveness of the Triops-induced morphological defences (i.e. an elongated tail spine and increased bulkiness) by conducting predation trials. 4. Our study provides rare evidence for morphological defences in D. magna, and in addition shows that prey species gradually switch between plastic traits to maintain effective defences throughout their entire lifetime. Hence, our results help to shed light on the mechanisms governing phenotypic plasticity within natural populations.

Constitutive and inducible defensive traits in co‐occurring marine snails distributed across a vertical rocky intertidal gradient

Summary 1. Among‐species differences in the expression of constitutive and plastic traits may shape species distributions within a community. For many systems, the relationships among individual species’ phenotypes, particularly morphological plasticity, and performance and distribution are poorly understood. 2. I tested whether three co‐occurring species of marine snails in the genus Nucella differed in their constitutive and inducible defences against crabs, in terms of growth and shell architectural defence. I also tested whether among‐species trait differences were associated with interspecific differences in antipredator performance (resistance to crushing) and distributions along a vertical tidal emersion gradient on rocky shores where the snails co‐occur. 3. Species distributed lower on the shore and thus exposed to crabs for longer periods of time exhibited higher constitutive growth rates than the upper‐shore species. Lower‐shore species also exhibited greater inducible reductions in somatic growth and increases in shell thickening than the upper‐shore species. Plastic alterations of the scaling relationship between shell‐thickness and somatic growth (i.e. greater shell mass per body mass) enabled the two lower‐shore species to produce stronger, more crab‐resistant shells in the presence of crabs than their upper‐shore congener. 4. Results suggest that species‐specific differences in both constitutive and inducible shell defence and growth reflect growth/mortality tradeoffs that are associated with species’ vertical distributions along a predation risk gradient. Thus, adaptive differences in inducible morphological defenses should be considered along with more traditionally studied traits such as behaviour and constitutive morphological defenses, when examining traits that influence patterns of species distribution along environmental gradients.

Cue reliability, risk sensitivity and inducible morphological defense in a marine snail

Reliable cues that communicate current or future environmental conditions are a requirement for the evolution of adaptive phenotypic plasticity, yet we often do not know which cues are responsible for the induction of particular plastic phenotypes. I examined the single and combined effects of cues from damaged prey and predator cues on the induction of plastic shell defenses and somatic growth in the marine snail Nucella lamellosa. Snails were exposed to chemical risk cues from a factorial combination of damaged prey presented in isolation or consumed by predatory crabs (Cancer productus). Water-borne cues from damaged conspecific and heterospecific snails did not affect plastic shell defenses (shell mass, shell thickness and apertural teeth) or somatic growth in N. lamellosa. Cues released by feeding crabs, independent of prey cue, had significant effects on shell mass and somatic growth, but only crabs consuming conspecific snails induced the full suite of plastic shell defenses in N. lamellosa and induced the greatest response in all shell traits and somatic growth. Thus the relationship between risk cue and inducible morphological defense is dependent on which cues and which morphological traits are examined. Results indicate that cues from damaged conspecifics alone do not trigger a response, but, in combination with predator cues, act to signal predation risk and trigger inducible defenses in this species. This ability to "label" predators as dangerous may decrease predator avoidance costs and highlights the importance of the feeding habits of predators on the expression of inducible defenses.

An inducible morphological defence is a passive by-product of behaviour in a marine snail

Many organisms have evolved inducible defences in response to spatial and temporal variability in predation risk. These defences are assumed to incur large costs to prey; however, few studies have investigated the mechanisms and costs underlying these adaptive responses. I examined the proximate cause of predator-induced shell thickening in a marine snail (Nucella lamellosa) and tested whether induced thickening leads to an increase in structural strength. Results indicate that although predators (crabs) induce thicker shells, the response is a passive by-product of reduced feeding and somatic growth rather than an active physiological response to predation risk. Physical tests indicate that although the shells of predator-induced snails are significantly stronger, the increase in performance is no different than that of snails with limited access to food. Increased shell strength is attributable to an increase in the energetically inexpensive microstructural layer rather than to material property changes in the shell. This mechanism suggests that predator-induced shell defences may be neither energetically nor developmentally costly. Positive correlations between antipredator behaviour and morphological defences may explain commonly observed associations between growth reduction and defence production in other systems and could have implications for the evolutionary potential of these plastic traits.

Evaluating inducible morphological defences in the common freshwater ciliate, Coleps hirtus

Experiments examined whether the ciliate, Coleps hirtus, changed its morphology in the presence of zooplankton known to prey on ciliates. While caudal spine length was not affected, Coleps were longer and thinner in the presence of Cyclops than in their absence.

Predator-induced changes in morphology of a prey fish: the effects of food level and temporal frequency of predation risk

In a series of experiments, we investigated the effects of food availability and risk frequency on the dynamics of predator-induced changes in growth and morphology of prey fish using goldfish (Carassius auratus) as our test species. In experiment 1, we fed goldfish high or low food rations and exposed them to either alarm cues from conspecifics, cues from swordtails or a water control. After 60 days, goldfish in the alarm cue treatment significantly increased their body depth and body weight but had smaller body length than goldfish exposed to swordtails cues or water, likely reducing their vulnerability to gape-limited predators. Importantly, food level had an impact on the amplitude of the morphological changes. In experiment 2, goldfish were exposed to two different frequencies of predation cues or a water control for 50 days. The cues were either continued or discontinued from day 51 to 100, and all cues were resumed from day 101 to 150. We found that goldfish exposed to predation cues increased their depth and weight at a faster rate than did the goldfish exposed to water, and of particular significance was the fact that frequency of risk had an effect on the amplitude of the change. When the cues were interrupted, the increase in growth rate parameters was reduced to the level of the goldfish exposed to water. However, when the cues were resumed, the rate increased to match the growth rate of the goldfish that were continuously exposed to the cues. Finally, we staged encounters between goldfish of differing morphologies and yellow perch (Perca flavescens) and found that deep-bodied goldfish had better survival than the shallow-bodied ones. These experiments illustrate the dynamic nature of inducible morphological defences.

Flexible inducible morphological defense in anuran larvae

Flexible inducible morphological defense in anuran larvae