Defenses In Daphnia Research Articles

One Kairomone and Multiple Effects in Daphnia Species—5α-Cyprinol Sulfate Induces Morphological Defenses in the Invasive Species Daphnia lumholtzi

Recently, the first chemical cues, which induce defenses in freshwater zooplankton of the genus Daphnia have been identified. Still it is unknown how general these so-called kairomones are and if they are perceived by and thereby benefitting invasive species. We here report the chemical identity of the fish-derived chemical signal that induces morphological defensive structures in the invasive species Daphnia lumholtzi. We used bioassay-guided isolation (LC-MS) of incubation water of fish and identified the bile salt 5α-cyprinol sulfate (5α-CPS) as the kairomone. We confirmed this finding by bioassays, in which D. lumholtzi was exposed to purified 5α-CPS sulfate from fish bile and showed that 10 pM 5α-CPS induces significantly elongated helmets and spines in relation to body length. These results identify on one hand another Daphnia species besides Daphnia magna, which is responding to this particular fish-borne kairomone, and on the other hand they reveal another defense that the kairomone induces besides diel vertical migration (DVM). Taken together a high evolutionary conservancy of the molecular mechanism behind inducible defenses in Daphnia spp. against fish can be deduced. It is reasonable to expect that 5α-CPS is involved in further predator—prey communication between Daphnia and fish, which may be of special ecological relevance with regard to invasive species as D. lumholtzi.

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.

Predator-specific inducible morphological defenses of a water flea against two freshwater predators.

The expression of inducible morphological defenses in Daphnia in response to a single predator is a well-known phenomenon. However, predator-specific modifications of the same defensive traits as an adaption to different predator regimes is so far only described for Daphnia barbata. It is unknown if this accounts only for this species or if it is a more widespread, general adaptive response in the genus Daphnia. In the present study, we therefore investigated whether a clone of the pond-dwelling species Daphnia similis responds to different predatory invertebrates (Triops cancriformis; Notonecta maculata) with the expression of predator-specific modifications of the same defensive traits. We showed that Triops-exposed individuals express a significantly longer tail-spine, while body width decreased in comparison to control individuals. Additionally, they also expressed inconspicuous defenses, that is, significantly longer spinules on the dorsal ridge. The Notonecta-exposed D. similis showed a significantly longer tail-spine, longer spinules and a larger spinules bearing area on the dorsal ridge than control individuals as well. However, a geometric morphometric analysis of the head shape revealed significant, predator-specific changes. Triops-exposed individuals expressed a flattened head shape with a pronounced dorsal edge, while Notonecta-exposed individuals developed a high and strongly rounded head. Our study describes so far unrecognized inducible defenses of D. similis against two predators in temporary waters. Furthermore, the predator-dependent change in head shape is in concordance with the 'concept of modality', which highlights the qualitative aspect of natural selection caused by predators.

Identification of Chaoborus kairomone chemicals that induce defences in Daphnia.

Infochemicals play important roles in aquatic ecosystems. They even modify food web interactions, such as by inducing defenses in prey. In one classic but still not fully understood example, the planktonic freshwater crustacean Daphnia pulex forms specific morphological defenses (neckteeth) induced by chemical cues (kairomones) released from its predator, the phantom midge larva Chaoborus. On the basis of liquid chromatography, mass spectrometry, and chemical synthesis, we report here the chemical identity of the Chaoborus kairomone. The biologically active cues consist of fatty acids conjugated to the amino group of glutamine via the N terminus. These cues are involved in Chaoborus digestive processes, which explains why they are consistently released despite the disadvantage for its emitter. The identification of the kairomone may allow in-depth studies on multiple aspects of this inducible defense system.

Rising pCO2 in Freshwater Ecosystems Has the Potential to Negatively Affect Predator-Induced Defenses in Daphnia

Anthropogenically released CO2 accumulates in the global carbon cycle and is anticipated to imbalance global carbon fluxes [1]. For example, increased atmospheric CO2 induces a net air-to-sea flux where the oceans take up large amounts of atmospheric CO2 (i.e., ocean acidification [2-5]). Research on ocean acidification is ongoing, and studies have demonstrated the consequences for ecosystems and organismal biology with major impacts on marine food webs, nutrient cycles, overall productivity, and biodiversity [6-9]. Yet, surprisingly little is known about the impact of anthropogenically caused CO2 on freshwater systems due to their more complex biogeochemistry. The current consensus, yet lacking data evidence, is that anthropogenic CO2 does indeed affect freshwater carbon hydrogeochemistry, causing increased pCO2 in freshwater bodies [10-13]. We analyzed long-term data from four freshwater reservoirs and observed a continuous pCO2 increase associated with a decrease in pH, indicating that not only the oceans but also inland waters areaccumulating CO2. We tested the effect of pCO2-dependent freshwater acidification using the cosmopolite crustacean Daphnia. For general validity, control pCO2-levels were based on the present global pCO2 average. Treatments were selected with very high pCO2 levels, assuming a continuous non-linear increase of pCO2, reflecting worst-case-scenario future pCO2 levels. Such levels of elevated pCO2 reduced the ability of Daphnia to sense its predators and form adequate inducible defenses. We furthermore determined that pCO2 and not the resulting reduction in pH impairs predator perception. If pCO2 alters chemical communication between freshwater species, this perturbs intra- and interspecific information transfer, which may affect all trophic levels.

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.

Predator-induced defences in Daphnia longicephala: location of kairomone receptors and timeline of sensitive phases to trait formation.

ABSTRACTThe freshwater crustacean Daphnia adapts to changing predation risks by forming inducible defences. These are only formed when they are advantageous, saving associated costs when the defence is superfluous. However, in order to be effective, the time lag between the onset of predation and the defence formation has to be short. Daphnia longicephala develop huge protective crests upon exposure to chemical cues (kairomones) from its predator the heteropteran backswimmer Notonecta glauca. To analyse time lags, we determined kairomone-sensitive stages and the developmental time frames of inducible defences. Moreover, we looked at additive effects that could result from the summation of prolonged kairomone exposure. Kairomones are perceived by chemoreceptors and integrated by the nervous system, which alters the developmental program leading to defence formation. The underlying neuronal and developmental pathways are not thoroughly described and surprisingly, the location of the kairomone receptors is undetermined. We show that D. longicephala start to sense predator cues at the onset of the second juvenile instar, defences develop with a time lag of one instar and prolonged kairomone exposure does not impact the magnitude of the defence. By establishing a method to reversibly impair chemosensors, we show the first antennae as the location of kairomone-detecting chemoreceptors. This study provides fundamental information on kairomone perception, kairomone-sensitive stages, developmental time frames and lag times of inducible defences in D. longicephala that will greatly contribute to the further understanding of the neuronal and developmental mechanisms of predator-induced defences in Daphnia.

Coupling of anti-predator defences in Daphnia: the importance of light

Here we test for the possible coupling of two kairomone-induced, anti-fish defences in Daphnia, life-history changes (LHC) and diel vertical migration (DVM) mediated by the environmental factor light. A gradient of five different light intensities that represents naturally occurring intensities in the lake water column was used in life-history experiments, and we show that LHC of a single Daphnia clone are inversely coupled to the ambient light intensity. Furthermore, we could show that the light intensity has to exceed a threshold to induce the LHC. We also observed an effect of the light intensity on the fish kairomone-mediated expression of a candidate gene (actin 3) in a way that the gene response differs between groups of low and high light intensities. We argue that the ambient light intensity an individual is exposed to and that is dependent on the position in the water column of a lake inversely determines the degree of LHC. These findings suggest a plastic coupling of these two anti-fish defences in Daphnia (LHC and DVM), which allows for an adjustment to fluctuating environments of standing freshwater ecosystems.

Proteomic analysis of Daphnia magna hints at molecular pathways involved in defensive plastic responses.

BackgroundPhenotypic plasticity in defensive traits occurs in many species when facing heterogeneous predator regimes. The waterflea Daphnia is well-known for showing a variety of these so called inducible defences. However, molecular mechanisms underlying this plasticity are poorly understood so far. We performed proteomic analysis on Daphnia magna exposed to chemical cues of the predator Triops cancriformis. D. magna develops an array of morphological changes in the presence of Triops including changes of carapace morphology and cuticle hardening.ResultsUsing the 2D-DIGE technique, 1500 protein spots could be matched and quantified. We discovered 179 protein spots with altered intensity when comparing Triops exposed animals to a control group, and 69 spots were identified using nano-LC MS/MS. Kairomone exposure increased the intensity of spots containing muscle proteins, cuticle proteins and chitin-modifying enzymes as well as enzymes of carbohydrate and energy metabolism. The yolk precursor protein vitellogenin decreased in abundance in 41 of 43 spots.ConclusionIdentified proteins may be either directly involved in carapace stability or reflect changes in energy demand and allocation costs in animals exposed to predator kairomones. Our results present promising candidate proteins involved in the expression of inducible defences in Daphnia and enable further in depth analysis of this phenomenon.

Ecological genomics: steps towards unraveling the genetic basis of inducible defenses in Daphnia

Little is known about the genetic mechanisms underlying inducible defenses. Recently, the genome of Daphnia pulex, a model organism for defense studies, has been sequenced. Building on the genome information, recent preliminary studies in BMC Developmental Biology and BMC Molecular Biology have assessed gene response profiles in Daphnia under predation pressure. We review the significance of the findings and highlight future research perspectives.See research articles http://www.biomedcentral.com/1471-2164/10/527, http://www.biomedcentral.com/1471-2105/6/45, http://www.biomedcentral.com/1471-213X/10/45

Turning inducible defenses on and off: adaptive responses of Daphnia to a gape‐limited predator

The use of inducible defenses is a common strategy to reduce predation while minimizing associated costs for prey. The most effective use of these defenses, however, may involve turning them on and off at different stages of ontogenetic development, with the timing dependent on prey body size and the nature of the predation environment. We develop a model based on the strike efficiency of a size-selective predator that examines the interaction between induced morphological defenses and prey body size, including the consequences of this interaction for the optimal development of the defenses during the prey's ontogeny. We then examine this model with respect to a model system of inducible defenses: neck spine induction in the water flea Daphnia in response to predatory larvae of the phantom midge Chaoborus. In accordance with predictions of the model, the body size and timing of neck spine acquisition during Daphnia development are related to the relative sizes of the Daphnia and Chaoborus species interacting in a pond or lake. The Daphnia species examined first acquire neck spines in either the first, second, or third juvenile instar, at body lengths that range from 0.58 to 0.83 mm. Neck spine formation is initiated at larger Daphnia body sizes when these prey are subject to predation by a larger Chaoborus species (C. trivittatus) and at smaller sizes when exposed only to a smaller predator (C. americanus). Induction of these morphological defenses in Daphnia occurs later in juvenile development in the smaller of the two species we examined (D. minnehaha) than in the larger (D. pulex). Delayed acquisition of neck spines also occurs when Daphnia are exposed to predation by larger Chaoborus. The close match between model predictions and the patterns observed in nature suggests that these patterns are adaptive developmental responses to different predator environments.

Inducible defences in Daphnia depend on latent alarm signals from conspecific prey activated in predators.

Some water fleas (Daphnia spp.) undergo phenotypic changes when exposed to chemical signals from predators. The chemical signals have been assumed to be of predator origin (i.e. kairomones), since juices of crushed Daphnia have been found ineffective. We speculated that latent alarm signals could be present in Daphnia, to be activated in predators following ingestion. Accordingly, fish predators were fed earthworms for 10 weeks to remove Daphnia remains from their gastro-intestinal tracts. Following another 6 days of earthworm feeding, water conditioned by fish induced no morphological changes in D. galeata. When fish were alternatively fed Daphnia for 6 days, changes were induced with fish-conditioned water. Extracts made from intestines of earthworm-fed fish, homogenized with earthworms, gave no morphological changes, but intestines of the same origin homogenized with Daphnia did. Similar results were found when earthworms and Daphnia were homogenized with fish liver. Freshly frozen extracts of homogenized Daphnia gave no detectable changes at first instar stage in test animals, whereas extracts of Daphnia that had been kept at room temperature did induce such changes. Our results suggest that Daphnia respond to latent conspecific alarm signals (i.e. 'dormant' pheromones) that are activated by intestinal or bacterial enzymes in predators or in the water.

Inducible defences in Daphnia: responses to two closely related predator species.

The predator-induced responses of two species of Australian Daphnia, with contrasting distributions and life history patterns, to the kairomones of two species of Anisops predators, were measured. Daphnia longicephala produced a large crest and attained a larger size when exposed to both predators. D. carinata sl matured earlier than D. longicephala and did not produce a crest. Surprisingly, kairomones of both predators inhibited the production of ephippia in D. carinata sl. Anisops stali, the larger of the two predator species, induced a significantly larger crest size in D. longicephala, and larger brood size in both species compared with the smaller A. gratus, indicating a quantitative but not qualitative effect of predator species on inducible defences.

Anti-Predator Defenses in Daphnia: Are Life-History Changes Always Linked to Induced Neck Spines?

The cladoceran Daphnia pulex responds phenotypically to chemicals released by one of its invertebrate predators Chaoborus flavicans. They produce neck spines with often several teeth which possibly increase the chance to survive the predator's attacks. On the other hand, these spined animals suffer a reduced fitness in terms of delayed maturity and reduced fecundity. Several studies describe both neck teeth and life-history responses as trade-offs which are directly linked although there is no direct evidence that neck teeth themselves bear those life-history costs. One possible way of testing the connection between morphology and life-history shifts is described in this study