Sperm number assessed by flow cytometry in species of Daphnia (Crustacea,Cladocera)

Daphnia magna and Daphnia pulex are two important model species in ecotoxicology. In daphniids, studies of the effects of contaminants have mostly focused on female life history traits, yet it would also be important to examine male reproductive traits, particularly in relation to endocrine disruptors. In this study, we developed a protocol that uses flow cytometry to measure sperm number in individual males of different species of Daphnia. We tested our protocol on 114 males from several clones of three common species of Daphnia. Sperm count varied widely among individuals and reached high numbers (up to 1.45 × 10⁵). Positive relationships between male length and sperm number were observed in D. pulex and Daphnia pulicaria, but not in D. magna. Important inter‐clonal differences in sperm production were observed in all species, with some clones producing very little sperm. Duplicated sperm samples showed on average only 6% difference in sperm counts. Sperm counts were stable at least over a 2‐hr period and up to 5 hr for most samples. This sperm isolation protocol and flow cytometric enumeration approach will be of major interest to ecotoxicologists.     

Coupling of growth rate and body stoichiometry in Daphnia: a role for maintenance processes?

1. The growth rate hypothesis predicts positive relationships among growth rate (μ), body RNA (%RNA of dry mass) and body P (%P of dry mass) contents. 2. We tested this within‐ and across‐species by growing five species/clones of Daphnia (Daphnia magna, Daphnia pulex, Daphnia galeata and two isolates of Daphnia pulicaria) with different combinations of food quantity and stoichiometric food quality. 3. Within each species, positive correlations among μ, %RNA and %P were seen and across species there was a strong association between%RNA and %P, consistent with the growth rate hypothesis. However, coupling of growth to %RNA and to %P differed for different species. In particular, the %RNA–μ and %P–μ relationships had similar slopes but considerably different y‐intercepts (i.e.%P or %RNA at zero growth), with D. pulicaria and D. galeata having higher intercepts than D. magna and especially D. pulex. As a result of these displacements, the relative rankings of the species on the basis of %P and %RNA did not correspond to their rankings based on μ. 4. These findings suggest that within a narrow clade (e.g. the daphnids), interspecific differences in body P content may reflect not growth rate‐related RNA allocation but instead the amount of RNA required for support of maintenance processes.     

Reproductive allocation in Daphnia exposed to toxic cyanobacteria

We investigated experimentally how resources were allocated to reproduction in Daphnia pulex and Daphnia longispina when varying levels of toxic Microcystis were added to higher quality food. We used multiple regression models to estimate mean offspring size and clutch size in relation to maternal size and clutch number, and analysed effects of treatments on residuals from the models. We also measured variation in per offspring investment. At a high cyanobacterial level, D.pulex was virtually unable to reproduce. At a lower level, D.pulex produced small clutches. However, the regression model residuals indicated that the presence of cyanobacteria increased the portion of available resources allocated to reproduction. The observed allocation may be a means to maximize reproduction under diminished longevity. Effects on mean offspring size were marginal in D.pulex but variation in per offspring investment sometimes decreased in cyanobacterial exposures. Daphnia longispina was affected by a higher cyanobacterial level only, where offspring sized was reduced. Deviations from the regression model indicated that effects on maternal size alone do not explain this effect. Clutch size residuals and per offspring investment were unaffected by treatments in D.longispina. The observed responses differ from theoretical models on reproductive allocation under food imitation.      

Characteristics of six cladocerans in relation to ecotoxicity testing

Investigation was made to compare some biological characteristics relevant to ecotoxicity testing among six cladoceran species, including Daphnia magna, Daphnia carinata, Daphnia pulex, Ceriodaphnia quadrangular, Bosmina longirostris, and Simocephalus vetulus. The results show that D. carinata had advantages over other cladoceran species for being used as a test organism, particularly for ecotoxicological assessment of aquatic environments in tropical and subtropical areas. D. carinata had similar body size and total number of offspring per female to D. magna. However, D. carinata was more sensitive to the reference toxicant and had much shorter reproduction cycle than D. magna. D. carinata had similarity to D. pulex, C. quadrangular, S. vetulus and B. longirostris in terms of sensitivity to the reference toxicant and length of reproduction cycle. However, D. carinata was much larger in size and produced much more offspring per female than any of D. pulex, C. quadrangular, S. vetulus and B. longirostris. Among the investigated cladocerans, only the neonates (2- and 4-day-old) of D. carinata exhibited phototaxis that was sufficiently remarkable. The low among-generation variation in phototaxis index (I_p) of D. carinata and the close relationship between I_p and the concentration of the ISO standard toxicant (K₂Cr₂O₇) appears to suggest that I_p can be used as an excellent test endpoint for ecotoxicity testing.     

Foraging strategies and growth inhibition in five daphnids feeding on mixtures of a toxic cyanobacterium and a green alga

1. Laboratory experiments were used to study the feeding, growth and reproduction of five daphnids in mixtures of a toxic cyanobacterium, Microcystis aeruginosa, and a green alga, Scenedesmus acutus. The mixtures included 0%, 20%, 50%, 80% and 100% Microcystis with a total food concentration of 0.5 mg C L^??1 in each treatment. The feeding rate was measured after 1 and 24 h of acclimatization to the mixtures. 2. Toxic Microcystis inhibited feeding in all the species, but they exhibited an unexpected diversity and complexity in the pattern of feeding inhibition. Daphnia magna exhibited the strongest inhibition of feeding after 1 h of exposure to toxic food, but had substantially recovered after 24 h in the same mixtures. This pattern of inhibition and recovery may balance the benefits of reduced ingestion of toxin with the disadvantage of a reduced energy intake. 3. All five daphnids grew quickly in the Scenedesmus control, whereas growth and reproduction declined with an increasing proportion of the toxic alga in the diet. Daphnia pulicaria showed the least inhibition of growth and reproduction, D. pulex showed the strongest inhibition and the three remaining species exhibited intermediate sensitivity. 4. Estimates of gross growth efficiency (GGE; growth/ingestion) provided a means for discriminating between the effects of feeding inhibition and direct toxicity on zooplankton growth. Daphnia pulex exhibited a sharp decline in GGE, suggesting that growth inhibition was a result of both feeding inhibition and direct toxicity. In contrast, D. magna exhibited a nearly constant GGE, indicating that feeding inhibition accounted for its decline in growth. However, two Daphnia species (i.e. D. pulicaria and D. galeata) exhibited improbable increases in GGE with toxic cyanobacteria, suggesting that their feeding rates were underestimated. Growth assays with sensitive and resistant zooplankton species are proposed for testing the potential impacts toxic cyanobacteria in lakes.     

The importance of social dimension and maturation stage for the probabilistic maturation reaction norm in Poecilia reticulata

Maturation is an important event in an organism's life history, with important implications on dynamics of both wild and captive populations. The probabilistic maturation reaction norm (PMRN) has emerged as an important method to describe variation in maturation in wild fish. Because most PMRNs are based on age and size only, it is important to understand limitations of these variables in explaining maturation. We experimentally assessed (i) the sensitivity of age‐ and size‐based PMRNs to unaccounted sources of plasticity, (ii) the role of social environment on maturation and (iii) the significance of estimating PMRNs early and late in the maturation process (initiation and completion of maturation, respectively). We reared male guppies (Poecilia reticulata) under laboratory conditions, subjected to two food levels and three different social cues. We found that growth and social environment affected the maturation in a way that could not be accounted for by their effect on age and size. PMRNs estimated for the initiation stage were less plastic (growth differences and social cues influenced the PMRN shape only little) than those for completion. The initiation of maturation is probably closer to the maturation ‘decision’ and allows determining factors influencing maturation decision most accurately.     

Notes on the systematics and ecology of Daphnia pulex LEYDIG in Northern Canada

Daphnia populations from the region of Yellowknife (N. W. T.) on the Great Nave Lake, from Tuktoyaktuk (N. W. T.) on the coast of the Beaufort Sea east of the Mackenzie-Delta, and from Shingle Point (Y. T.) west of the Mackenzie-Delta were described. Population diagrams were given for the beginning of September 1974. The populations found at the above mentioned sites were compared with Daphnia pulex from Spitsbergen and the German island of Spiekeroog. Though some characteristics of Daphnia middendorffiana FISCHER occurred, mainly in individuals from very northern ponds, these were always found in individuals which also possessed some of the characteristics of Daphnia pulex. In every pond, specimens with characteristics of Daphnia middendorffiana lived together with other individuals resembling Daphnia pulex. Males were found in three out of the four visited ponds on the shore of the Arctic Sea, and were apparently Daphnia pulex. All the material sampled in northern Canada was regarded as belonging to Daphnia pulex LEYDIG 1860 emend. RICHARD 1896. There was evidence, that characteristics of Daphnia middendorffiana develop under the influence of environmental conditions, such as food supply. Daphnia magna STRAUS was found far north of the arctic circle.     

Comparative sensitivity of the several zooplankton species (Cladocera,Copepoda) to sumicidine-alpha insecticide

The sensitivity of seven zooplankton species (Daphnia magna, D. longispina, D. pulex, Simocephalus vetulus, Scapholeberis mucronata, Eudiaptomus graciloides, and Cyclops strenuus) to an insecticide, sumicidine alpha was studied. LD₅₀ values upon a 24-hour exposure were determined. Daphnia pulex and D. longispina turned out to be highly susceptible to the insecticide, while the individuals representing the natural population of D. magna happened to be the most resistant. The data on sensitivity distribution among different zooplankton species are provided, as well as the concentrations potentially hazardous for 5% and 50% of the population are calculated.     

Molecular genetic identification and phylogeny of Daphnia species (Crustacea, Cladocera) from water bodies of the Lake Chany basin

The data on the molecular genetic identification of Daphnia species from the water bodies of the Lake Chany basin are presented. Phylogenetic relationships between these species have been established. The fragments of the mitochondrial DNA genes were used as genetic markers. According to the data obtained, the water bodies examined were inhabited by five Daphnia species, including Daphnia (Daphnia) galeata Sars, D. (D.) longispina O. F. Müller, D. (D.) curvirostris Eylmann, D. (D.) pulex Leydig, and D. (Ctenodaphnia) magna Straus. In addition, longispina a-like individuals that form a separate phylogenetic lineage was identified.     

Temperature and the effects of elemental food quality on Daphnia

1. We examined the responses of two species of Daphnia to changes in food phosphorus (P) content, with animals reared at three different water temperatures. Specifically, we measured mass‐specific growth rate (MSGR), body P content and respiration rate of Daphnia magna and Daphnia pulex acclimatised to 10, 17.5 and 25 °C and fed food carbon : phosphorus (C : P) ratios of either 150 or 500. 2. The responses of these three physiological variables to temperature–food quality interactions were species‐specific. There was a significant interactive effect of temperature and food quality on D. magna, as the greatest proportional effect of food quality on growth was observed at 10 °C and reductions in body P because of low food P content were relatively greater at 25 °C. These effects may reflect the temperature dependence of mechanisms that reduce elemental constraints associated with food quality in D. magna. By contrast, there were no interactive effects between food quality and temperature on MSGR, body P or mass‐specific respiration of D. pulex. 3. It thus appears that temperature can alter food quality effects on Daphnia but the nature of these alterations depends upon the daphniid species and its thermal adaptability. Significant temperature–food quality interactions will complicate efforts to understand zooplankton nutrition in nature and warrant future consideration.     

Life history responses of Daphnia clones to toxic Microcystis at different food levels

We examined how the amount of food of good quality, Scenedrsmusobtusiusculus, influenced the toxic effects of the cyanobacteriumMicrocystu aeruginosa on Daphnia pulex in a 21 day life tableexperiment. Our results show that all life history traits studied,i.e. longevity, growth and reproduction, of D.pulex were negativelyaffected by toxic Microcystis. The detrimental effects on theselife history traits were less pronounced at increasing foodlevels. In addition, D.pule.x clones differed in their lifehistory responses to toxic Microcystis, suggesting that thepresence of toxic cyanobacteria may modify the outcome amongD.pulex clones.     

Neck spine protects Daphnia pulex from predation by Chaoborus, but individuals with longer tail spine are at a greater risk

We tested the prey preference of Chaoborus sp. on Daphnia pulexwith different defensive morphologies. The protective functionof inducible morphological defences, such as neck spine andlonger tail spine, was evaluated Second instar D.pulex individualsof two clones differing in their strength of neck spine inductionwere offered as prey to both Chaoborus obscuripes and Chaoborusflavicans. We used logistic regression analysis to evaluatethe effect of morphometry on the vulnerability of Daphnia. Thepresence of a neck spine and increased total length protectedD.pulex from Chaoborus predation. However, individuals witha longer tail spine were more vulnerable to Chaoborus predationChaoborus obscuripes was able to eat daphnids with a neck spinedue to the larger gape size of this chaoborid. The smaller speciesC.flavicans almost always ate prey with no neck spine.     

Resource-niche complementarity and autotrophic compensation determines ecosystem-level responses to increased cladoceran species richness

This study examines the relationship between cladoceran species richness and ecosystem functioning. I conducted an experiment in which four cladocerans, Daphnia. magna, D. longispina, D. pulex and Chydorus sphaericus, were cultured in microcosms using different species combinations and levels of species richness. The results demonstrate that even within this closely related group of organisms the effects on ecosystem-level variables, such as total algae and zooplankton biomass, per capita productivity, and nutrient concentrations, as well as phytoplankton community structure, were highly variable between different combinations of these species. Since only four species where involved in this study, species-specific effects dominated the general relationship between species richness and ecosystem functioning. Particular combinations of species resulted in effects that indicated more efficient grazing. These effects, which were most pronounced in combinations including both D. magna and C. sphaericus, were manifested as an indirect effect as the prey community shifted towards grazing-resistant species. As a result, the productivity of the prey community decreased, because phytoplankton species with lower per capita productivity became more dominant. I suggest that the primary mechanism that caused this significant effect was complementarity in prey-size use of D. magna and C. sphaericus. In terms of prey-size range, D. pulex and D. longispina were redundant when D. magna was present and were quickly out-competed by the latter despite higher per capita filtering efficiency. The results show that different mechanisms are important for different combinations of species. Furthermore, the ability of the prey community to respond to changes of consumer species composition is an important factor in experiments in which consumer species richness is experimentally manipulated. Received: 2 November 1998 / Accepted: 8 September 1999     

Intraspecific variability in response to stimuli for male and ephippia formation in Daphnia pulex

Natural clones and clones from laboratory crossings of Daphnia pulex have been tested in their response to stimuli for male and ephippia formation. The clones are from ponds in Illinois, USA, an area with both obligate and cyclical clones of D. pulex. The progeny from two types of crosses were studied: 1) crosses between an obligate clone producing males and a cyclical clone producing only females, and 2) self-fertilization within a cyclical clone producing both sexes. Both the natural and the artificial clones showed great variation in response to environmental stimuli for males. Ephippia response seems more equal among the clones, but two of the artificial clones could hardly be stimulated to produce ephippia in the induction experiments. In crowded cultures, however, they produced a few. Crossings between cyclicals and obligates yielded mostly cyclical progeny. Progeny from the selfed clone had low survival and fecundity.     

Mitochondrial DNA diversity in an apomictic Daphnia complex from the Canadian High Arctic

Cyclic parthenogenesis is the ancestral mode of reproduction in the cladoceran crustacean, Daphnia pulex, but some populations have made the transition to obligate parthenogenesis and this is the only mode of reproduction known to occur in arctic populations. Melanism and polyploidy are also common in arctic populations of this species. Prior allozyme studies of arctic D. pulex revealed substantial levels of clonal diversity on a regional scale. Clonal groupings based on cluster analysis of allozyme genotypes do not conform to groupings based on the presence/absence of melanin or on ploidy level. In order to further elucidate genetic relationships among arctic D. pulex clones, mitochondrial DNA (mtDNA) variation was examined in 31 populations from two Canadian high-arctic sites. The data were also compared to a previous study of mtDNA variation in populations from a Canadian low-arctic site. Cladistic analysis of restriction site variation of the entire mitochondrial genome and nucleotide sequence variation of the mitochondrial control region was used to construct genetic relationships among mitochondrial genotypes. Three distinct mitochondrial lineages were detected. One lineage was associated with diploid, nonmelanic clones and is the same as the lineage that is found in temperate populations of D. pulex. The other two lineages (A & B) were associated with polyploid, melanic clones. Sequence divergence between the A and B lineages was 2.4%. Sequence divergence between D. pulex and either of these two lineages exceeded 3%. It is suggested that the melanic, polyploid clones are hybrids between males of D. pulex (and/or a closely related congener, D. pulicaria) and females of either of two ancestral melanic species that have mitochondrial lineages A and B. Geographic patterns of mitochondrial diversity in ‘melanic’ lineage B support the hypothesis of an high-arctic refuge for the ancestral species during the last glacial period.     

Karyologic observations on the maturation of the summer and winter Eggs of Daphnia pulex and Daphnia middendorffiana

The cladoceran Daphnia middendorffiana is an arctic species able to produce winter eggs which develop parthenogenetically. The cytological study of the maturation of summer and winter eggs of an italian population of this species has shown that both types of eggs undergo only one maturation division of equational type with the expulsion of only one diploid polar body, the same maturation process as had been found in the pseudosexual eggs of a race of Daphnia pulex (Schrader, 1925).—The chromosomes behaviour during the maturation of summer and winter eggs of D. middendorffiana has been compared with that of a population of D. pulex which shows a normal heterogonic cycle.This investigation was supported by the Consiglio Nazionale delle Ricerche (C.N.R.) of Italy.     

Variation in life history responses of Daphnia to toxic Microcystis aeruginosa

Three clones of Daphnia pulex and two clones of Daphnia longispinawere exposed to toxic Microcystis aeruginosa for 21 days ina lifetable experiment. The growth and reproduction of individualdaphnids were followed daily to study the long-term effectsof toxic Microcystis. Exposure to Microcystis increased mortality,decreased growth, delayed maturation and decreased offspringproduction, indicating nutritional deficiency and toxic effects.We found variation in life history responses between speciesand among clones. Our results suggest that toxic cyanobacteriamay act as a modifying agent in zooplankton communities at boththe species and clonal level.     

Oddity,predation risk and social decisions in aquatic invertebrates

Group living is widespread across animal taxa, incurring benefits such as increased foraging efficiency or an enhanced chance of surviving a predator's attack. The chances of escaping a predator are often lower for odd‐looking individuals, as these are detected at a higher rate than uniform looking group members. While this “oddity effect” shall operate in animals differing in any given phenotype, including colour, size or species identity, it has been experimentally tested mainly for odd‐coloured individuals. We examined the oddity effect and swarming preferences in two differently sized species of freshwater planktonic crustaceans (large Daphnia magna and small Daphnia pulex). We experimentally investigated whether odd individuals in a swarm of heterospecific Daphnia were more vulnerable to predation by a common predator, the three‐spined stickleback (Gasterosteus aculeatus). Furthermore, Daphnia's swarming preference was tested by giving individuals the opportunity to choose between conspecific/heterospecific odour and a neutral control. In contrast to the predictions of the oddity effect, odd individuals were not always preyed on earlier; instead three‐spined stickleback preferentially predated large, more nutritious individuals. Daphnia of both species reacted towards the perception of con‐ and heterospecifics odours. While D. pulex generally avoided the smell of other daphnids, D. magna avoided conspecifics but tended to prefer heterospecifics over the neutral control. These findings provide new insights into swarming strategies and social preference of an invertebrate and how this behaviour can influence predation risk.     

Is competition important to arctic zooplankton community structure?

1. Daphnia pulex and Daphnia middendorffiana are commonly found in the Toolik Lake region of arctic Alaska. These two species are very similar morphologically, although their natural distributions differ markedly: D. pulex is restricted to shallow ponds, while D. middendorffiana is widely distributed and found in a variety of ponds and lakes. We compared the reproductive capabilities of D. pulex and D. middendorffiana grown under similar resource conditions and in the absence of the invertebrate predator Heterocope septentrionalis. In situ life table and mesocosm experiments were conducted in Toolik Lake and Dam Pond, habitats that have historically contained natural populations of D. middendorffiana, but never D. pulex. 2. Daphnia pulex exhibited a significantly higher net growth rate than D. middendorffiana in both life table and mesocosm experiments although D. pulex has never been found in either Toolik Lake or Dam Pond. Daphnia middendorffiana exhibited a negative net growth rate in Dam Pond, which had lower resource levels then Toolik Lake. Therefore, the smaller D. pulex appears to have a lower food threshold concentration than the larger D. middendorffiana. 3. Our results indicate that D. pulex is a superior resource competitor in the Toolik Lake region. These results combined with distributional patterns suggest that the restricted distribution of D. pulex in these arctic lakes and ponds cannot be explained by resource competition alone. We suggest that in the presence of H. septentrionalis, predation is an important factor structuring arctic zooplankton communities in the Toolik Lake region.