Waterborne metals impair inducible defences in Daphnia pulex: morphology, life-history traits and encounters with predators

1.  Inducible defences may be temporary and favoured where predation is intermittent and have been demonstrated in several invertebrates and vertebrates when prey detect chemical cues (kairomones) released by predators. Daphnia pulex (a water flea) exposed to Chaoborus (midge larvae) kairomones produce small neckteeth on the dorsal surface of the head as a defence against this gape-limited predator and survive better in the presence of Chaoborus . Recent studies have shown that waterborne copper (Cu) impairs the induction of neckteeth which could lead to lower survival.
2.  Here, we examined the effects of Cu on morphological changes and shifts in life-history traits in D. pulex exposed to kairomone from Chaoborus americanus . We exposed D. pulex mothers to chemical cues of C. americanus fed on either D. pulex neonates or on brine shrimp Artemia salina , the same Chaoborus cues combined with an environmentally relevant concentration of copper (10 μg L⁻¹), or dechlorinated tap water. We examined several morphological characteristics of neonates and life-historical characteristics of adults as well as assessing survival of neonates by staging encounters with predators.
3.  Neonates from mothers exposed to kairomone plus copper had fewer and shorter neckteeth than neonates from mothers exposed to kairomone alone. Moreover, neonates exposed to Cu had lower survival during encounters with predators than neonates exposed to kairomone without Cu.
4.  Adult female Daphnia exposed to kairomones released more neonates within the first 24 h of brood release and emptied their brood pouches quicker than mothers not exposed to kairomones, irrespective of the presence of Cu.
5.  Impairment by metals of morphological defences in zooplankton could lead to a decline in population density and alter community structure.     

Antipredator reaction norms for life history traits in Daphnia pulex: dependence on temperature and food

Synergistic effects between temperature and food level on the vulnerability of Daphnia life histories to predation have previously received little attention, despite their potential significance for summer population dynamics. In this investigation, most traits in the early life history of Daphnia pulex altered in response to increasing concentrations of fish kairomone. Although there was some variation attributable to experimental temperature and food treatments, traits exhibited reaction norms which suggested at least some degree of kairomone dose-dependence. Temperature, food level and their interaction affected the mean values of every trait examined and in some cases also influenced the antipredator response via a three-way interaction with fish kairomone. The insertion of an extra juvenile instar resulted in later maturation at a larger size for most females raised under conditions of low temperature and low food, but this response was largely suppressed in the presence of fish kairomone. Earlier maturation due to deletion of an instar was observed most frequently under high food and temperature conditions with fish kairomone present, suggesting an effect of kairomone on the size threshold for reproductive development. Principal components analysis was used to produce an index reflecting the net potential vulnerability to fish predation of the suite of life history traits. Vulnerability generally declined with increasing kairomone level as a result of the apparently adaptive alterations in most life history traits. Raised temperature and food level also generally reduced potential vulnerability, but a highly significant interaction between these factors was also found. Potentially important implications of these results for optimal vertical migration and summer population dynamics are discussed.     

The effect of temperature on the response of Daphnia to toxic cyanobacteria

1. In a set of life‐table experiments, cohorts of neonate Daphnia pulex were exposed to a toxic strain of Anabaena affinis and A. flos ‐ aquae , and to a pure cyanobacterial toxin (anatoxin‐a), at 12 or 14, 19, and 25 °C. The fecundity and survival of individual animals were assessed at 1‐, 2‐ or 3‐day intervals, depending on the temperature, through to the fifth brood of the control cohort. The sensitivity of D . pulex to the cyanobacteria and the toxin at each temperature was measured by determining its finite population growth rate (λ) in an experimental treatment as a fraction of that in a control treatment. Tests with three concentrations of cyanobacteria (1.0, 2.5 and 5.0 μg mL^–1) and one concentration of anatoxin‐a (1 μg mL^–1), and with two clones of D. pulex , showed a consistent and statistically significant pattern of increasing sensitivity at higher temperatures.
2. Anabaena affinis affected both survivorship and fecundity, while A. flos ‐ aquae and its toxin, anatoxin‐a, primarily affected fecundity. Presence of cyanobacteria affected brood size, brood number, time to first reproduction and interclutch interval. Temperature affected time to first reproduction and interclutch interval at all concentrations of cyanobacteria. Brood number and brood size were little affected by temperature except at the highest concentrations of cyanobacteria. Increasing the concentration of A. flos ‐ aquae affected demographic parameters, especially at the lower temperatures, while increasing the concentration of A. affinis had less effect.
3. The study suggests that increasing water temperatures in natural systems should exacerbate the inhibitory effect of toxic cyanobacteria on daphniid population growth rates.     

Predator–prey instability: individual-level mechanisms for population-level results

1. In previous work we established that increasing temperature led to a destabilization of the population dynamics of the invertebrate carnivore Mesostoma ehrenbergii and its prey Daphnia pulex , which ultimately resulted in the local extinction of Daphnia at higher temperatures. Two mechanisms are proposed to explain the population-level phenomena: (1) quantitative changes in carnivore vital rates with increasing temperature led to stronger functional and numerical response and (2) qualitative changes in the dynamic allocation of energy to reproduction by the predator with increasing temperature introduces inverse density dependence in the predator's response.
2. The growth of individual M. ehrenbergii was monitored under various food conditions to determine the effect of two temperatures (18 and 24 °C) and five food levels on rates of growth, prey consumption and reproduction and on reproductive allocation patterns.
3. The first mechanism was supported by both higher consumption rates (stronger functional response) and faster growth rates with earlier age at maturity and shorter generation time (stronger numerical response).
4. Evidence for mechanism two was also provided by an alteration of the reproductive allocation pattern with temperature. Viviparous (subitaneous) eggs were more likely to be produced by this carnivore at low food levels at 24 °C, while at 18 °C, high food levels were required before individuals made this switch. This shift actually introduces inverse density dependence in the predator's numerical response which is highly destabilizing.
5. Based on the results of this study, the differential effect of M. ehrenbergii on the dynamics and structure of its D. pulex prey populations can be attributed to changes in both physiological rates and reproductive allocation patterns with temperature.     

Effects of low pH and humus on the survivorship, growth and feeding of Gammarus pulex (L.) (Amphipoda)

SUMMARY. 1. The toxicity to the freshwater amphipod Gammarus pulex (L.) of soft water (pH6.0) with three concentrations of XAD-extracted aquatic humus was treated in the laboratory.
2. Exposure to pH6.0 water without humus added resulted in 92% mortality after 3 weeks. Humus, added at concentrations of 7 and 20 g Cm ³, decreased the mortality to 80% and 64% in the same period.
3. Surviving animals kept at pH6.0 had a lower growth rate, lower food conversion efficiency and higher body water content than animals kept at pH 7.3.
4. Humus had no significant effects on growth, food conversion or body composition of G. pulex kept at pH 6.0. However, there was a tendency for growth and food conversion to increase, and for body water content to decrease with increased humus concentration. The effects of humus on growth and food conversion of G. pulex observed in this study do not support the contention that humus acts as a free coupler to lower metabolic efficiency. On the contrary, humus tended to benefit food conversion.
5. It is suggested that low concentrations of humus can be directly beneficial to organisms in acidified water in ways other than by complexing toxic metals.     

Dietary essential amino acids affect the reproduction of the keystone herbivore Daphnia pulex

Recent studies have indicated that nitrogen availability can be an important determinant of primary production in freshwater lakes and that herbivore growth can be limited by low dietary nitrogen availability. Furthermore, a lack of specific essential nitrogenous biochemicals (such as essential amino acids) might be another important constraint on the fitness of consumers. This might be of particular importance for cladoceran zooplankton, which can switch between two alternative reproductive strategies--the production of subitaneously developing and resting eggs. Here, we hypothesize that both the somatic growth and the type of reproduction of the aquatic keystone herbivore Daphnia is limited by the availability of specific essential amino acids in the diet. In laboratory experiments, we investigated this hypothesis by feeding a high quality phytoplankton organism (Cryptomonas) and a green alga of moderate nutritional quality (Chlamydomonas) to a clone of Daphnia pulex with and without the addition of essential amino acids. The somatic growth of D. pulex differed between the algae of different nutritional quality, but not dependent on the addition of dissolved amino acids. However, in reproduction experiments, where moderate crowding conditions at saturating food quantities were applied, addition of the essential amino acids arginine and histidine (but not lysine and threonine) increased the total number and the developmental stage of subitaneous eggs. While D. pulex did not produce resting eggs on Cryptomonas, relatively high numbers of resting eggs were released on Chlamydomonas. When arginine and histidine were added to the green algal diet, the production of resting eggs was effectively suppressed. This demonstrates the high, but previously overlooked importance of single essential amino acids for the reproductive strategy of the aquatic keystone herbivore Daphnia.     

Derivation of a toxicity-based model to predict how water chemistry influences silver toxicity to invertebrates

The effect of altering water chemistry on acute silver toxicity to three invertebrate species, two Daphnids, Daphnia magna and Daphnia pulex, as well as an amphipod Gammarus pulex was assessed. In addition, the physiological basis of Ag(I) toxicity to G. pulex was examined. Daphnia magna and D. pulex were more sensitive than G. pulex and 48 h LC(50) values in synthetic ion poor water were 0.47, 0.65 and 2.1 microg Ag(I) l(-1), respectively. Increasing water [Cl(-)] reduced Ag(I) toxicity in all species, and increasing water [Ca(2+)] from 50 to 1,500 microM reduced Ag(I) toxicity in G. pulex. Whole body Na(+) content, but not K(+) or Ca(2+) was significantly reduced in G. pulex exposed to 6 microg Ag(I) l(-1) for 24 h, but there was no inhibition of whole body Na(+)/K(+)-ATPase activity. Both increasing water [Cl(-)] and [Ca(2+)] reduced this Ag(I)-induced Na(+) loss. For D. magna, the presence of 10 mg l(-1) humic acid or 0.5 microM 3-mercaptoproprionic acid (3-MPA) increased the 48 h LC(50) values by 5.9 and 58.5-fold, respectively, and for D. pulex the presence of 1 microM thiosulfate increased the 48 h LC(50) value by four-fold. The D. magna toxicity data generated from this study were used to derive a Daphnia biotic ligand model (BLM). Analysis of the measured LC(50) values vs. the predicted LC(50) values for toxicity data from the present and published results where water Cl(-), Ca(2+), Na(+) or humic acid were varied showed that 91% of the measured toxicity data fell within a factor of two of the predicted LC(50) values. However, the daphnid BLM could not accurately predict G. pulex toxicity. Additionally, the Daphnia BLM was under-protective in the presence of the organic thiols 3-MPA or thiosulphate and predicted an increase in the LC(50) value of 114- and 74-fold, respectively. The Daphnia toxicity based BLM derived from the present data set is successful in predicting Daphnia toxicity in laboratory data sets in the absence of sulfur containing compounds, but shows its limitations when applied to waters containing organic thiols or thiosulphate.     

The effects of food and temperature regimes on life-history responses to fish kairomones in Daphnia hyalina × galeata*

Life-history responses to two concentrations of fish released info-chemicals at two temperature and food regimes were investigated for one clone of Daphnia hyalina × galeata. The presence of fish kairomones had a negative impact on size at maturity, carbon allocation to individual eggs and size of neonates in all treatments. Food concentration and temperature had positive effects on size of adult stages, independent of kairomone treatment. However, kairomone treatment were not found to interact with food or temperature. Age at maturity was positively influenced by increased temperature and food concentrations, whereas no direct kairomone effects were detected for this trait. Clutch size was not directly influenced by kairomone treatment, whereas both food concentration and temperature had strong, positive effects.     

Endosulfan-enhanced crest induction in Daphnia longicephala: evidence for cholinergic innervation of kairomone receptors

Daphnia iongicephala were reared from early embryogenesis inthe presence or absence of crest-inducing kairomones releasedby Anisops gratus (Notonectidae) and from birth in one of fiveconcentrations of the organochiorine pesticide, endosulfan (0,0.1, 1.0, 10 or 100 g 1^–1 The morphologyof the daphnidswas measured 3 days after birth and on production of the firstbrood of eggs. The reproductive parameters, first brood size,age at first reproduction, mean egg volume and total egg volume,were also measured. Endosulfan significantly induced crest developmentin 3-day-old daphnids at 0.1, 1.0 and 10 g ^1– in thepresence and absence of Anisops kairomone, although crest sizein the absence of kairomone was only marginally greater thancontrols. A concentration of 10 pg 1^–1 endosulfan significantlyenhanced crest growth of kairomone-exposed daphnids at maturity.The Anisops kairomone alone induced a large crest in D.iongicephola,reduced first brood size, mean egg volume and total egg volume,and increased age at maturity. The coefficient of variationof mean egg volume was significantly reduced by Anisops kairomone.It is hypothesized that endosulfan enhances crest developmentbyinhibition of .     

Life history consequences for Daphnia pulex feeding on nutrient-limited phytoplankton

1. The growth and feeding of Daphnia pulex De Geer on different algal species was examined. The green algae Chlamydomonas reinhardtii Dangeard and Scenedesmus acutus Meyen, the diatom Synedra tenuissima Kützing, the cryptophyte Cryptomonas pyrenoidifera Geitler and the cyanobacterium Microcystis aeruginosa Kützing were cultured in non-limiting and in N- or P-limiting medium and used as food for D. pulex.
2. Growth limitations were reflected in the elemental and biochemical composition and the morphological characteristics of the algal resources.
3. The clearance rates of D. pulex feeding on nutrient-limited algae were reduced. This was not observed when nutrient-limited mutant Chlamydomonas cells without cell walls were used as food, indicating that the cell wall may play an important part.
4. In comparison with animals grown on nutrient-sufficient cells, nutrient-limited algae resulted in smaller body length, reduced brood sizes, reduced size at maturity, increased age at first reproduction and, consequently, in reduced Daphnia population growth rates.
5. Daphnia population growth rates ( r ) were negatively correlated with the C : P ratio and the carbohydrate content of the food. Moreover, significant correlations between r and clearance rates were found.
6. The observed differences in the grazing and the life history parameters of Daphnia feeding on non-limited and nutrient-limited algae may be the result of both reduced nutritional value and reduced digestibility of nutrient-limited algae.     

Variation in the neonate size of Daphnia pulex: the effects of predator exposure and clonal origin

Daphnia pulex clones originating from twelve small pond habitats were exposed to chemical cues from a size-selective predator, larvae of the phantom midge Chaoborus. Exposure delayed the onset of reproduction and increased the size at first reproduction. On the other hand, the neonates produced by these larger mothers were slightly smaller than the neonates produced by the smaller control mothers. In cladocerans, neonate size is usually positively correlated to the size of the mother. Thus exposure to Chaoborus kairomone apparently had direct effects on neonate size counterbalancing the maternal effects. Daphnia clones collected from Chaoborus-free and Chaoborus-rich environments exhibited different responses. In first adult instar, the clones from Chaoborus inhabited environments increased their offspring size under Chaoborus exposure whereas clones from Chaoborus-free environments did not. This may reflect clonal adaptation to the predation prehistory of their original habitat since larger neonates more quickly reach a size protected from the predator.     

Genome size evolution and polyploidy in the Daphnia pulex complex (Cladocera: Daphniidae)

Genome size was estimated in 49 clones of the Daphnia pulex complex from temperate and subarctic locations using flow cytometry and microsatellite DNA analyses. Significant genome size differences were found in diploid species belonging to the two genetically distinct groups (the pulicaria and the tenebrosa groups), with clones from the tenebrosa group having genome sizes 22% larger than those in the pulicaria group. Combined flow cytometry and microsatellite DNA analyses revealed that nearly all polyploid clones in the D. pulex complex are triploid and not tetraploid, as was previously suggested. Sequencing analyses of the ND5 gene to position clones in their respective clades within the D. pulex complex have uncovered three triploid clones of Daphnia middendorffiana with a D. pulex maternal parent. This result was unexpected because Daphnia pulicaria has always been identified as the maternal parent of these hybrid polyploid clones. Triploid clones likely owe their origins to interactions between sexual and asexual populations. Further interactions in the tenebrosa group have generated tetraploid clones but these events have been rare.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 68–79.     

Salamander predation and vertical distributions of zooplankton

SUMMARY. 1. The impact of different predation regimes of zooplanktivorous larval tiger salamanders, Ambystoma tigrinum nebulosum , on the vertical temporal distributions of Daphnia pulex, Diaptomus nudus and larval Chaoborus flavicans was studied in limnetic enclosures in a pond in east-central Arizona. Zooplankton densities were monitored 1-2 h before sunset, at sunset, and 1-2 h after sunset at three depths (0-0.4, 1.0 and 2.0 m) in four salamander treatments (0, 20, 40 salamanders per enclosure, and unenclosed limnetic areas).
2. Densities and body size of Daphnia pulex were consistently greater in salamander-free enclosures than in other treatments. More Daphnia were generally nearer the surface (0-1.0 m) than along the bottom before and at sunset than after sunset in enclosures without salamanders. After sunset, Daphnia was by far most dense along the bottom. Densities were uniform among depths and sample times in treatments with salamanders. Salamanders moved throughout the water column. Frequencies of D. pulex spined morphs (with prominent dorsal crest) did not differ among salamander treatments.
3. Chaoborus densities were positively correlated with D. pulex distributions among depths and sample times in treatments. Spatio-temporal distributions of Chaoborus were influenced more by their daphnid prey than by risk of predation from larval salamanders.
4. Densities of Diaptomus nudus were highest in open water controls and lowest in enclosures without salamanders. Diaptomus was generally more dense between 0 and 1 m than at 2.0 m depths in all treatments, and vertical distributions were not dependent on salamander density.     

Temperature effects on chemical signalling in a predator–prey system

SUMMARY 1. We investigated the effect of temperature on chemical signalling in a predator–prey model system (planktivorous fish and Daphnia galeata ). Life-history changes in Daphnia in response to chemical cues (kairomones) derived from fish have become a paradigm for chemically induced anti-predator defences.
2. As temperature can affect both predator and prey, we carried out two experiments to disentangle these effects. In order to test for temperature effects on the predator, we kept prey at a single temperature and exposed them to kairomones from fish exposed to two different temperatures. Daphnia exhibited a higher intrinsic rate of population increase ( r ) when exposed to fish kairomones produced at high rather than low temperature. Assuming a positive correlation between r (because of an earlier maturation and/or increased clutch sizes) and kairomone concentration, our results suggest that kairomone production increases with rising temperature.
3. In the second experiment, to study the influence of temperature on the prey, Daphnia were kept at two different temperatures and exposed to fish kairomones produced at one constant temperature. We found no interaction between the effects of fish kairomone and temperature on Daphnia life history, suggesting that temperature does not directly alter life-history responses to fish kairomones.
4. Our results suggest that temperature influences Daphnia life history through its effects on fish kairomone concentration, but that temperature does not affect the strength of the response of Daphnia to the presence of fish.     

Phenotypic plasticity of Daphnia life history traits: the roles of predation, food level and toxic cyanobacteria

1. We studied the life history responses of Daphnia pulex under different biotic conditions. In a factorially designed experiment, we tested the impacts of water conditioned by the invertebrate predator Chaoborus, low and high food level (10 000 and 100 000 Scenedesmus cells ml^–1), and exposure to toxic Microcystis (5000 cells ml^–1) on twelve D. pulex clones originating from different habitats. Our aim was to compare the phenotypic plasticity of different clones, and to study the interactions among biotic factors.
2. Individuals cultured in Chaoborus -conditioned water started to reproduce at a larger size than individuals cultured in water not conditioned by the predators. We found interactions between food level and Chaoborus -conditioned water on age at first reproduction and total offspring number. In addition, the impact of the Chaoborus treatment on the size at first reproduction was reduced by Microcystis exposure.
3. Clonal differences were found in the degree of phenotypic plasticity of different life history traits. However, there was no obvious connection between the original habitat of the clone and the degree of plasticity.     

Sexual dimorphism in Gammarus pulex: the effect of current flow on pre-copula pair formation

SUMMARY. 1. Male Gammarus pulex (L.) are typically twice as heavy as females in field caught pre-copula pairs and mating is positively size assortative. Earlier work had shown that relatively large males can swim in pre-copula at higher current speeds than males that are relatively small compared with their mates. We describe experiments on pair formation in the laboratory, in static water and in an artificial stream.
2. Pairs formed in the artificial stream had a significantly higher weight ratio than either those formed in static water or those caught in the field. In both experimental conditions, males in pairs were significantly larger than singles. In the stream, the weight ratio of pairs increased with formation sequence due to a significant decrease in the size of selected females.
3. It is concluded that these results are consistent with the hypothesis that the relatively large size of paired male G. pulex is a product of mechanical constraints rather than male–male competition.     

The temperature‐size rule in Daphnia magna across different genetic lines and ontogenetic stages: Multiple patterns and mechanisms

Ectotherms tend to grow faster, but reach a smaller size when reared under warmer conditions. This temperature‐size rule (TSR) is a widespread phenomenon. Despite the generality of this pattern, no general explanation has been found. We therefore tested the relative importance of two proposed mechanisms for the TSR: (1) a stronger increase in development rate relative to growth rate at higher temperatures, which would cause a smaller size at maturity, and (2) resource limitation placing stronger constraints on growth in large individuals at higher temperatures, which would cause problems with attaining a large size in warm conditions. We raised Daphnia magna at eight temperatures to assess their size at maturity, asymptotic size, and size of their offspring. We used three clonal lines that differed in asymptotic size and growth rate. A resource allocation model was developed and fitted to our empirical data to explore the effect of both mechanisms for the TSR. The genetic lines of D. magna showed different temperature dependence of growth and development rates resulting in different responses for size at maturity. Also, at warm temperatures, growth was constrained in large, but not in small individuals. The resource allocation model could fit these empirical data well. Based on our empirical results and model explorations, the TSR of D. magna at maturity is best explained by a stronger increase in development rate relative to growth rate at high temperature, and the TSR at asymptotic size is best explained by a size‐dependent and temperature‐dependent constraint on growth, although resource limitation could also affect size at maturity. In conclusion, the TSR can take different forms for offspring size, size at maturity, and asymptotic size and each form can arise from its own mechanism, which could be an essential step toward finding a solution to this century‐old puzzle.     

Does trimethylamine induce life-history reactions in Daphnia?

Laboratory experiments were carried out to investigate life-history reactions of Daphnia to trimethylamine (TMA), a substance, which has recently been found to induce a similar phototactic reaction in Daphnia as fish kairomones. The effects of different treatments (control, fish kairomone and TMA) on the life-history traits of five clones of D. magna and one clone of D. galeata and D. hyalina, respectively, were studied. Only D. magna exhibited significant reactions to TMA and to fish kairomone. D. galeata showed no significant responses to either TMA or to fish kairomone. D. hyalina reacted to the fish treatment with two traits (size and number of eggs at maturity). The comparison of the reaction norms to TMA and fish kairomone demonstrated that the directions of some responses to both factors were similar but were different for others such as for size at maturity. TMA resulted in a larger size at maturity in D. magna, whereas fish kairomone had no significant influence on this trait. We, therefore, conclude that trimethylamine is not the primary causative chemical agent in fish induced life-history adaptations in Daphnia.     

The influence of changing temperature on the life history of Daphniopsis ephemeralis

We present evidence which suggests that for Daphniopsis ephemeralischanges in water temperature may directly cue changes in sexratio, ephippial and parthenogenetic egg production and indirectlydetermine female body size. Field data suggested that over anannual cycle, a population of D. ephemeralis comprised animalswith three distinct life history phases. The first was madeup of exephippial females which hatched in the fall. They produceda second group made up of large parthenogenetic females whichcould produce more than one overwintering generation. In thespring these parthenogenetic females gave rise to a third lifehistory phase which comprised males and small ephippial females.Field enclosure experiments suggested that the small size ofspring females was not related to predation. Laboratory experimentssuggested that ephippial egg production at small body sizeswas associated with ‘increasing’ temperature. Theseexperiments also suggested that the fall reproductive pattern(parthenogenetic eggs and large body sizes) was associated with‘decreasing’ temperatures. We conclude that directionalityof seasonal temperature change may be the prime factor responsiblefor D. ephemeralis life history changes observed in the field.     

Shortened lifespan: another cost of fish-predator avoidance in cladocerans?

Life-history theory predicts adaptive shifts in response to size-selective predation, namely earlier reproduction, smaller age/size at maturity, and higher relative investment into reproduction. Such shifts should bring about reduced lifespan of potential prey. We tested this prediction in life-table experiments with clones of Daphnia hyalina and Diaphanosoma brachyurum, two species of contrasting anti-predatory strategies. The clones were derived from seven lakes of different trophy and held in water with and without fish kairomone, under standard laboratory conditions. Exposure to the kairomone caused a decrease in age of first reproduction and an increase in early-life reproductive effort but also an about 20% decrease of longevity in both species. Although shortened lifespan did not result in significant decrease in fitness of the tested species (in terms of lifetime reproductive output) it should be taken into account in considerations of costs and benefits of inducible defenses in cladocerans.