Does a Diet Rich in the Bacterium Rhodopirellula rubra Improve Daphnia magna Performance?

BACKGROUND: In the wild various organisms contribute to daphnids diet. This study, intendeds to evaluate the potential of the concentration of Rhodopirellula rubra as a single or supplementary food source for Daphnia magna. METHODS: Feeding assays were performed according to standard guidelines for chronic assays (21 days), and life-history parameters and several biomarkers (protein content, oxidative stress, energetic reserves and pigments) were measured. Five food regimens were conducted with 20 individual replicates (A - R. subcapitata; 0.2 - suspension of R. rubra at 0.2 arbitrary units (AU); 0.4 - suspension of R. rubra at 0.4 AU; 0.2+A - suspension of R. rubra at 0.2+alga; 0.2+A-suspension of R. rubra at 0.4 AU + alga). Additionally, the effects of three diets (A, 0.2, and 0.2+A) on the longevity of D. magna were assessed. RESULTS: The five diets showed a different C, N, and carotenoids composition, with an increase in the mixed diets. The results confirmed that the mixed diets improved D. magna life-history parameters. A decrease in glycogen, and the increase of haemoglobin, protein, and gluthione-S-transferase (GST) were observed. Furthermore, D. magna fed with bacterial single diets, presented worsen life history parameters and a decrease in the protein content. An induction of oxidative stress response (increased catalase and GST), and a significant decrease in lipid peroxidation and an accumulation of glycogen and carotenoids were observed. Overall, an increase in the amount of R. rubra provided to D. magna, from 0.2 AU to 0.4 AU, negatively impacted daphnid performance. No significant effects on Daphnia longevity (a 110-day assay) were observed among the three diets tested. However, a significant survival percentage and fertility (cumulative offspring is more than twice) was observed when D. magna was fed with the mixed diet. CONCLUSIONS: Results demonstrated that different diets provided a nutritional diversified food to the daphnids that induced differences in D. magna performance. The mixed diets proved to be beneficial (with increase in offspring) on D. magna performance, independently of the bacterial concentration tested. When in single diet, bacterial concentration is not nutritionally sufficient to raise D. magna even when in increased concentration.

Proteome changes in an aquatic invertebrate consumer in response to different nutritional stressors.

Nutrient imbalances in zooplankton are caused by the differences in elemental content of producers and the demand for elements in consumers, which alter the life-history traits in consumers. Changes in life-history traits are mediated through metabolic pathways that affect gene expression and the metabolome. However, less is known about proteomic changes to elemental-limitation in zooplankton. Here, we grew Daphnia pulex under high food quantity and quality (HF), low food quantity (LF), and phosphorus (P)-limited (PL) diets for six days and measured growth, elemental composition, and the proteome. Daphnids in both LF and PL diets grew less. Animals in LF diets had less carbon (C), while daphnids in PL diets had less P compared to HF fed animals. In total, we identified 1719 proteins that were used in a partial least squares regression discriminant analysis (PLS-DA). Focusing on a subset of the proteome, the PLS-DA resulted in a clear separation between animals fed HF diets and PL and LF diets. Many proteome changes in nutrient-limited diets are associated with growth, reproduction, lipid metabolism, and nutrient assimilation. Regardless of the limiting nutrient, there were less hemoglobin and small subunit processome component proteins compared to HF fed animals. Daphnids fed LF diets had less vitellogenin fused superoxide dismutase and more lipid-droplet hydrolase, whereas Daphnia fed PL diets had higher abundances of cytochrome P450 and serine protease. Our proteome results compliment other "omic" studies that could be used to study Daphnia physiology in lakes.

Uranium accumulation and toxicokinetics in the crustacean Daphnia magna provide perspective to toxicodynamic responses.

The importance of incorporating kinetic approaches in order to gain information on underlying physiological processes explaining species sensitivity to environmental stressors has been highlighted in recent years. Uranium is present in the aquatic environment worldwide due to naturally occurring and anthropogenic sources, posing a potential risk to freshwater taxa in contaminated areas. Although literature shows that organisms vary widely with respect to susceptibility to U, information on toxicokinetics that may explain the variation in toxicodynamic responses is scarce. In the present work, Daphnia magna were exposed to a range of environmentally relevant U concentrations (0 - 200 µg L-1) followed by a 48 h depuration phase to obtain information on toxicokinetic parameters and toxic responses. Results showed time-dependent and concentration-dependent uptake of U in daphnia (ku = 1.2 - 3.8 L g-1 day-1) with bioconcentration factors (BCFs) ranging from 1,641 - 5,204 (L kg-1), a high depuration rate constant (ke = 0.75 day-1), the majority of U tightly bound to the exoskeleton (~ 50 - 60%) and maternal transfer of U (1 - 7%). Effects on growth, survivorship and major ion homeostasis strongly correlated with exposure (external or internal) and toxicokinetic parameters (uptake rates, ku, BCF), indicating that uptake and internalization drives U toxicity responses in D. magna. Interference from U with ion uptake pathways and homeostasis was highlighted by the alteration in whole-body ion concentrations, their ionic ratios (e.g., Ca:Mg and Na:K) and the increased expression in some ion regulating genes. Together, this work adds to the limited data examining U kinetics in freshwater taxa and, in addition, provides perspective on factors influencing stress, toxicity and adaptive response to environmental contaminants such as uranium.

Performance of Daphnia magna on flour, leaves, and pollen from different maize lines: Implications for risk assessment of genetically engineered crops.

Non-target effects of genetically engineered (GE) plants on aquatic Daphnia magna have been studied by feeding the species with different maize materials containing insecticidal Cry proteins from Bacillus thuringiensis (Bt). The results of those studies were often difficult to interpret, because only one GE plant was compared to one related non-GE control. In such a setting, effects of the Cry proteins cannot be distinguished from plant background effects, in particular when the test species is nutritionally stressed. In the present study, we tested the suitability of three different maize materials, i.e., flour, leaves and pollen, from five diverse non-GE maize lines (including EXP 258, a breeding line that is closely related to a SmartStax Bt maize) as exclusive food sources for D. magna. The parameters recorded included survival, sublethal endpoints such as body size, number of moltings to first offspring, time to first offspring, number of individuals in first clutch, total number of clutches, total number of offspring, average number of offspring per clutch, and population measures such as net reproductive rate R0, generation time T and intrinsic rate of increase rm. The results showed that D. magna can survive, grow and reproduce when fed only maize materials, although the performance was poorer than when fed algae, which indicates nutritional stress. Large differences in life table and population parameters of D. magna were observed among the different maize lines. Our results suggest that confounding effects caused by nutritional stress and plant background might explain some of the conflicting results previously published on the effects of Bt crops on D. magna. Using 95% confidence intervals for the means of the five maize lines for all measured parameters of D. magna performance in our study, we captured the natural range of variation. This information is useful for the interpretation of observed differences in D. magna performance between a GE plant and its non-GE comparator as it helps judging whether observed effects are of biological relevance. If differences between a GE and comparator line are observed and their biological relevance needs to be assessed in future risk assessments of GE maize, 1) the data on natural variation of the different parameters generated by previous studies can be informative (e.g. data from our study for maize fed D. magna); 2) for additional experiments the inclusion of multiple unrelated non-GE comparators should be considered; In addition, it should be taken into account that nutritional stress can affect the outcome of the study.

The complexity of co-limitation: nutrigenomics reveal non-additive interactions of calcium and phosphorus on gene expression in Daphnia pulex.

Many lakes across Canada and northern Europe have experienced declines in ambient phosphorus (P) and calcium (Ca) supply for over 20 years. While these declines might create or exacerbate nutrient limitation in aquatic food webs, our ability to detect and quantify different types of nutrient stress on zooplankton remains rudimentary. Here, we used growth bioassay experiments and whole transcriptome RNAseq, collectively nutrigenomics, to examine the nutritional phenotypes produced by low supplies of P and Ca separately and together in the freshwater zooplankter Daphnia pulex. We found that daphniids in all three nutrient-deficient categories grew slower and differed in their elemental composition. Our RNAseq results show distinct responses in singly limited treatments (Ca or P) and largely a mix of these responses in animals under low Ca and P conditions. Deeper investigation of effect magnitude and gene functional annotations reveals this patchwork of responses to cumulatively represent a co-limited nutritional phenotype. Linear discriminant analysis identified a significant separation between nutritional treatments based upon gene expression patterns with the expression patterns of just five genes needed to predict animal nutritional status with 99% accuracy. These data reveal how nutritional phenotypes are altered by individual and co-limitation of two highly important nutritional elements (Ca and P) and provide evidence that aquatic consumers can respond to limitation by more than one nutrient at a time by differentially altering their metabolism. This use of nutrigenomics demonstrates its potential to address many of the inherent complexities in studying interactions between multiple nutritional stressors in ecology and beyond.

Rearing Temperature and Fatty Acid Supplementation Jointly Affect Lipid Fluorescence Polarization and Heat Tolerance in Daphnia.

The homeoviscous adaptation hypothesis states that the relative abundance of polyunsaturated fatty acids (PUFAs) in membrane phospholipids of ectothermic organisms decreases with increasing temperatures to maintain vital membrane properties. We reared Daphnia magna at 15°, 20°, and 25°C and increasing dietary concentrations of the long-chain PUFA eicosapentaenoic acid (EPA) to test the hypothesis that the well-documented increase in heat tolerance of high-temperature-reared Daphnia is due to a reduction in body PUFA concentrations. Heat tolerance was assessed by measuring the time to immobility at a lethally high temperature (Timm at 37°C), and whole body lipid fluorescence polarization (FP) was used as an estimate of membrane fluidity. At all rearing temperatures, EPA supplementation resulted in an increase in the relative abundance of EPA in body tissues, but only at 15° and 25°C did this result in a decrease in heat tolerance, and only at 20°C was this associated with an increase in membrane fluidity (i.e., decrease in FP). Overall, however, the degree of tissue fatty acid unsaturation correlated well with heat tolerance and FP. Our results support the homeoviscous adaptation hypothesis by showing that cold-reared Daphnia accumulate PUFAs within their body tissues and thus are more susceptible to heat than hot-reared Daphnia accumulating fewer PUFAs. However, our data also point out that further studies are required that elucidate the complex relationships between PUFA supply, membrane fluidity, and heat tolerance in ectotherms.

Toxicity of wastewater from health care facilities assessed by different bioassays.

OBJECTIVES: The purpose of this study was to determine toxicity of wastewater from hospitals in the Czech Republic using traditional and alternative toxicological methods. The pilot study comprised weekly dynamics of sewage ecotoxicity of treated wastewater from one hospital in two different seasons. A detailed investigation of wastewater ecotoxicity, genotoxicity and reprotoxicity followed in five different hospitals. METHODS: The seven following bioassays were used in this study: algal growth inhibition test (ISO 8692), Vibrio fischeri test (ISO 11348-2), Daphnia magna acute toxicity test (ISO 6341), Allium cepa assay, Ames test (OECD TG 471), Comet assay and YES/YAS assay. RESULTS: The wastewater ecotoxicity during one week showed no differences in separate working days, however, higher toxicity values were recorded in May compared to November. In the following study, samples from two of the five hospitals were classified as toxic, the others as non toxic. Genotoxicity has not been confirmed in any sample. In several cases, wastewater samples exhibited agonist activity to the estrogen and androgen receptors. CONCLUSION: The study demonstrated different levels of toxicity of treated hospital wastewater. Variable sensitivity of individual bioassays for tested wastewater samples was recognized. A more extensive study including proposal for improvement of hospital wastewater treatment within the Czech Republic can be recommended with the aim to decrease the discharge of toxic chemicals into the local sewage system and the environment.

Fish-mediated plankton responses to increased temperature in subtropical aquatic mesocosm ecosystems: Implications for lake management.

Although it is well established that climate warming can reinforce eutrophication in shallow lakes by altering top-down and bottom-up processes in the food web and biogeochemical cycling, recent studies in temperate zones have also shown that adverse effects of rising temperature are diminished in fishless systems. Whereas the removal of zooplanktivorous fish may be useful in attempts to mitigate eutrophication in temperate shallow lakes, it is uncertain whether similar mitigation might be achieved in warmer climates. We compared the responses of zooplankton and phytoplankton communities to climate warming in the presence and absence of fish (Aristichthys nobilis) in a 4-month mesocosm experiment at subtropical temperatures. We hypothesized that 1) fish and phytoplankton would benefit from warming, while zooplankton would suffer in fish-present mesocosms and 2) warming would favor zooplankton growth but reduce phytoplankton biomass in fish-absent mesocosms. Our results showed significant interacting effects of warming and fish presence on both phytoplankton and zooplankton. In mesocosms with fish, biomasses of fish and phytoplankton increased in heated treatments, while biomasses of Daphnia and total zooplankton declined. Warming reduced the proportion of large Daphnia in total zooplankton biomass, and reduced the zooplankton to phytoplankton biomass ratio, but increased the ratio of chlorophyll a to total phosphorus, indicating a relaxation of zooplankton grazing pressure on phytoplankton. Meanwhile, warming resulted in a 3-fold increase in TP concentrations in the mesocosms with fish present. The results suggest that climate warming has the potential to boost eutrophication in shallow lakes via both top-down (loss of herbivores) and bottom-up (elevated nutrient) effects. However, in the mesocosms without fish, there was no decline in large Daphnia or in total zooplankton biomass, supporting the conclusion that fish predation is the major driver of low large Daphnia abundance in warm lakes. In the fishless mesocosms, phytoplankton biomass and nutrient levels were not affected by temperature. Our study suggests that removing fish to mitigate warming effects on eutrophication may be potentially beneficial in subtropical lakes, though the rapid recruitment of fish in such lakes may present a challenge to success in the long-term.

A mechanism study on toxicity of graphene oxide to Daphnia magna: Direct link between bioaccumulation and oxidative stress.

Graphene oxide (GO) possesses versatile applicability and high hydrophilicity, thus may have frequent contact with aquatic organisms. However, the ecological risks of GO in aquatic ecosystems remain largely unexplored currently. This study evaluated the comprehensive toxicological effects of GO on Daphnia magna, a key species in fresh water ecosystem. The results revealed nonsevere acute toxicities, including immobility (72 h EC50: 44.3 mg/L) and mortality (72 h LC50: 45.4 mg/L), of GO on D. magna. To understand the underlying mechanism of GO exposure, changes in superoxide dismutase (SOD) and lipid peroxidation (LPO) of D. magna exposed to GO were correlated, which revealed elevated GO-mediated oxidative stress and damages, especially in the long-time and high-dose exposure groups. The observations of in vivo fluorescence labeled with 2', 7'-dichlorofluorescin further demonstrated that reactive oxygen species were concentrated in daphnia guts, which corresponded with the high bioaccumulation level (5 mg/L, 24 h body burden: 107.9 g/kg) of GO in daphnia guts. However, depuration of GO from daphnia was not difficult. Daphnia almost released all GO within 24 h after it was transferred to clean water. These results hence suggest that GO could accumulate and induce significant oxidative stress in the gut of D. magna, while D. daphnia can also relieve the acute toxicity by depurating GO.

Effect of excess food nutrient on producer-grazer model under stoichiometric and toxicological constraints.

Accurately assessing the risks of contaminants requires more than an understanding of the effects of contaminants on individual organism, but requires further understanding of complex ecological interactions, elemental cycling, and the interactive effects of natural stressors, such as resource limitations, and contaminant stressors. There is increasing evidence that organisms experience interactive effects of contaminant stressors and food conditions, such as resource stoichiometry, availability and excess of nutrient. Here, we develop and analyze the first producer-grazer population model that incorporates the effects of excess nutrients, as well as nutrient limitations on grazer exposed to toxicants. We use analytical, numerical and bifurcation analysis to reduce and exploremodel parameterized for an aquatic system of algae and zooplankton exposed to methylmercury under varying phosphorus conditions. Under certain environmental conditions, our models predict higher toxicity than previous models that neglect the consequences excess nutrient conditions can have on grazer populations.

Mechanistically derived spatially heterogeneous producer-grazer model subject to stoichiometric constraints.

Known stoichiometric models of a two species producer-grazer ecosystem have either neglected spatial dynamics or failed to track free phosphorus in the media. In this paper we present a spatially heterogeneous model that tracks phosphorus content in the producer and free phosphorus in the media. We simulate our model numerically under various environmental conditions. Multiple equilibria, with bistability and deterministic extinction of the grazer, are possible here. In conditions that had been previously studied without tracking free phosphorus we find cases where qualitatively different behavior is observed. In particular under certain environmental conditions previous models predict stable equilibria where our model predicts stable limit cycles near the surface. Oscillatory dynamics can have consequences on the population densities, which may spend some time at low values throughout the cycles where they are in danger of stochastic extinction.

A historical perspective of nutrient change impact on an infectious disease in Daphnia.

Changes in food quality can play a substantial role in the vulnerability of hosts to infectious diseases. In this study, we focused on the genetic differentiation of the water flea Daphnia magna towards food of different quality (by manipulating C:N:P ratios) and its impact on the interaction with a virulent infectious disease, "White Fat Cell Disease (WFCD)". Via a resurrection ecology approach, we isolated two Daphnia subpopulations from different depths in a sediment core, which were exposed to parasites and a nutrient ratio gradient in a common garden experiment. Our results showed a genetic basis for sensitivity towards food deprivation. Both fecundity and host survival was differently affected when fed with low-quality food. This strongly impacted the way both subpopulations interacted with this parasite. A historical reconstruction of nutrient changes in a sediment core reflected an increase in organic material and phosphorus concentration (more eutrophic conditions) over time in the studied pond. These results enable us to relate patterns of genetic differentiation in sensitivity towards food deprivation to an increasing level of eutrophication of the subpopulations, which ultimately impacts parasite virulence effects. This finding was confirmed via a dynamic energy budgets (DEB), in which energy was partitioned for the host and the parasite. The model was tailored to our study by integrating (1) increased growth and a fecundity shift in the host upon parasitism and (2) differences of food assimilation in the subpopulations showing that a reduced nutrient assimilation resulted in increased parasite virulence. The combination of our experiment with the DEB model shows that it is important to consider genetic diversity when studying the impact of nutritional stress on species interactions, especially in the context of changing environments and emerging infectious diseases.

Combined effects of dietary polyunsaturated fatty acids and parasite exposure on eicosanoid-related gene expression in an invertebrate model.

Eicosanoids derive from essential polyunsaturated fatty acids (PUFA) and play crucial roles in immunity, development, and reproduction. However, potential links between dietary PUFA supply and eicosanoid biosynthesis are poorly understood, especially in invertebrates. Using Daphnia magna and its bacterial parasite Pasteuria ramosa as model system, we studied the expression of genes coding for key enzymes in eicosanoid biosynthesis and of genes related to oogenesis in response to dietary arachidonic acid and eicosapentaenoic acid in parasite-exposed and non-exposed animals. Gene expression related to cyclooxygenase activity was especially responsive to the dietary PUFA supply and parasite challenge, indicating a role for prostanoid eicosanoids in immunity and reproduction. Vitellogenin gene expression was induced upon parasite exposure in all food treatments, suggesting infection-related interference with the host's reproductive system. Our findings highlight the potential of dietary PUFA to modulate the expression of key enzymes involved in eicosanoid biosynthesis and reproduction and thus underpin the idea that the dietary PUFA supply can influence invertebrate immune functions and host-parasite interactions.

Arachidonic acid enhances reproduction in Daphnia magna and mitigates changes in sex ratios induced by pyriproxyfen.

Arachidonic acid is 1 of only 2 unsaturated fatty acids retained in the ovaries of crustaceans and an inhibitor of HR97g, a nuclear receptor expressed in adult ovaries. The authors hypothesized that, as a key fatty acid, arachidonic acid may be associated with reproduction and potentially environmental sex determination in Daphnia. Reproduction assays with arachidonic acid indicate that it alters female:male sex ratios by increasing female production. This reproductive effect only occurred during a restricted Pseudokirchneriella subcapitata diet. Next, the authors tested whether enriching a poorer algal diet (Chlorella vulgaris) with arachidonic acid enhances overall reproduction and sex ratios. Arachidonic acid enrichment of a C. vulgaris diet also enhances fecundity at 1.0 µM and 4.0 µM by 30% to 40% in the presence and absence of pyriproxyfen. This indicates that arachidonic acid is crucial in reproduction regardless of environmental sex determination. Furthermore, the data indicate that P. subcapitata may provide a threshold concentration of arachidonic acid needed for reproduction. Diet-switch experiments from P. subcapitata to C. vulgaris mitigate some, but not all, of arachidonic acid's effects when compared with a C. vulgaris-only diet, suggesting that some arachidonic acid provided by P. subcapitata is retained. In summary, arachidonic acid supplementation increases reproduction and represses pyriproxyfen-induced environmental sex determination in D. magna in restricted diets. A diet rich in arachidonic acid may provide protection from some reproductive toxicants such as the juvenile hormone agonist pyriproxyfen. Environ Toxicol Chem 2015;34:527-535. © 2014 SETAC.

A dietary polyunsaturated fatty acid improves consumer performance during challenge with an opportunistic bacterial pathogen.

A dietary deficiency in polyunsaturated fatty acids (PUFAs) and/or sterols can severely constrain growth and reproduction of invertebrate consumers. Single nutrients are potentially assigned to different physiological processes, for example to support defence mechanisms; therefore, lipid requirements of healthy and pathogen-challenged consumers might differ. In an oral exposure experiment, we explored the effects of dietary PUFAs and cholesterol on growth, reproduction and survival of an aquatic key herbivore (Daphnia magna) exposed to an opportunistic pathogen (Pseudomonas sp.). We show that healthy and pathogen-challenged D. magna are strongly albeit differentially affected by the biochemical composition of their food sources. Supplementation of a C20 PUFA-deficient diet with arachidonic acid (ARA) resulted in increased survival and reproduction of pathogen-challenged D. magna. We propose that the observed benefit of consuming an ARA-rich diet during pathogen challenge is conveyed partially via ARA-derived eicosanoids. This study is one of the first to consider the importance of dietary PUFAs in modifying fitness parameters of pathogen-challenged invertebrate hosts. Our results suggest that dietary PUFA supply should receive increased attention in host-microorganisms interactions and invertebrate disease models to better understand and predict disease dynamics in natural populations.

Aquatic hazard and biodegradability of light and middle atmospheric distillate petroleum streams.

Light and middle atmospheric distillate petroleum substances are blended to produce fuels used in transportation and heating. These substances represent the majority by volume of crude oil refined products in the United States. The goal of this research was to develop biodegradability and aquatic toxicity data for four substances; heavy, straight-run naphtha (HSRN), hydro-desulfurized kerosene (HDK), hydro-cracked gas oil (HCGO), and catalytic-cracked gas oil (CCGO). Ready biodegradability tests demonstrated rapid and extensive microbial oxidation of these test substances, indicating a lack of persistence in the aquatic environment. Differences in biodegradation patterns reflected compositional differences in the constituent hydrocarbons. Results of aquatic toxicity tests on alga, cladocera, and fish demonstrated that toxicity was greatest for catalytic-cracked gas oil, which contained a high proportion of aromatic hydrocarbons. Aromatic hydrocarbons are more soluble, and hence more bioavailable, resulting in higher toxicity. When expressed on the basis of loading rates, acute toxicity values (LL/EL50) ranged between 0.3 and 5.5 mg L(-1) for all three species, while chronic no-observed-effect loading rates (NOELR) ranged between 0.05 and 0.64 mg L(-1). PETROTOX estimates for acute and chronic toxicity ranged from 0.18 to 2.3 mg L(-1) and 0.06 to 0.14 mg L(-1), respectively, which were generally more conservative than experimental data.

A new approach of Integrated Health Responses (IHR(s)) modeling for ecological risk/health assessments of an urban stream.

The objective of this study was to evaluate the ecological health of an urban stream using Integrated Health Responses (IHRs). Water chemistry analysis, habitat health, and ecotoxicity tests were conducted in the stream along with analyses of molecular/biochemical, physiological biomarkers, and population-level responses in indicator species. Chemical stresses, measured as nutrient levels, ionic content and organic matter concentrations were significantly greater (p<0.01) at the downstream than the reference site (RF). The habitat health was largely impacted in the downstream reaches and had a negative relation with the land-use pattern of % urban area. Comet assay, 7-ethoxyresorufin-O-deethylase (EROD), acetylcholinesterase (AChE), and vitellogenin (VTG) were evaluated for low-level biomarker responses on DNA/physiological conditions of target species. The multi-metric fish model (Mm-F) was used to test the community-level response in relation to chemical and physical habitat stresses. The impaired responses of separate biomarker and bioindicator at the downstream sites occurred at all organizations from molecular/biochemical level to community level. Using all biomarkers/bioindicators, the star-plot model of IHRs was developed and then the integrative health/risk assessments were conducted in the urban stream. The reduced values of IHRs occurred in the downstream sites and the impacts were attributed to effluents from wastewater treatment plants (WTPs) and industrial complex. Ecological health impairments, thus, were evident in the urban reach, and reflected the long-term community responses as well as short-term responses of molecular biomarkers. The degradation of the urban stream was mainly due to a combined effect of chemical pollution and physical habitat modifications.

Impact of imidacloprid on Daphnia magna under different food quality regimes.

Aquatic ecosystems are characterized by fluctuating conditions that have direct effects on aquatic communities but also indirect influences such as changing the toxicity of chemicals. Because the effect of food quality on pesticide toxicity has rarely been studied, in the present study Daphnia magna juveniles supplied with 4 different food quality levels were exposed to a range of imidacloprid concentrations for 21 d. Food quality was expressed as carbon:phosphorus ratios of algae Pseudokirchneriella subcapitata (C:P 35, C:P 240, C:P 400, and C:P 1300). Survival, growth rates, and reproduction of D. magna were monitored, and the combined effects of imidacloprid exposure and the phosphorus content of algae were analyzed. A stronger effect on survival was observed at the P-deficient diet (C:P 1300), confirmed by lower 10% effect concentration (EC10) values at days 7, 9, 15, and 21 compared with diets with higher phosphorus contents. Similarly, the growth rate was reduced when D. magna were supplied with algae of low phosphorus content at imidacloprid exposure conditions. The highest reproductive output was observed for D. magna fed the optimal phosphorus diet (C:P 240), both at control and exposed conditions. Poor food quality increased the sensitivity of nontarget species to pesticide exposure, potentially leading to an underestimation of adverse effects on aquatic communities in the field.

Could contaminant induced mutations lead to a genetic diversity overestimation?

Contaminant driven genetic erosion reported through the inspection of selectable traits can be underestimated using neutral markers. This divergence was previously reported in the aquatic system of an abandoned pyrite mine. The most sensitive genotypes of the microcrustacean cladoceran Daphnia longispina were found to be lacking in the impacted reservoir near the entrance of the metal rich acid mine drainage (AMD). Since that divergence could be, at least partially, accounted for by mutagenicity and genotoxicity of the AMD, the present study aimed at providing such a characterization. The Allium cepa chromosomal aberration assay, using root meristematic cells, was carried out, by exposing seeds to 100, 10, 1, and 0.1 % of the local AMD. Chromosomal aberrations, cell division phases and cell death were quantified after the AMD exposure and after 24 and 48 h recovery periods. The AMD revealed to be mutagenic and genotoxic, even after diluting it to 1 and 0.1 %. Dilutions within this range were previously found to be below the lethality threshold and to elicit sublethal effects on reproduction of locally collected D. longispina clonal lineages Significant mutagenic effects (micronuclei and chromosomal breaks) were also found at 0.1 % AMD, supporting that exposure may induce permanent genetic alterations. Recovery tests showed that AMD genotoxic effects persisted after the exposure.

The good, the bad and the plenty: interactive effects of food quality and quantity on the growth of different Daphnia species.

Effects of food quality and quantity on consumers are neither independent nor interchangeable. Although consumer growth and reproduction show strong variation in relation to both food quality and quantity, the effects of food quality or food quantity have usually been studied in isolation. In two experiments, we studied the growth and reproduction in three filter-feeding freshwater zooplankton species, i.e. Daphnia galeata x hyalina, D. pulicaria and D. magna, on their algal food (Scenedesmus obliquus), varying in carbon to phosphorus (C∶P) ratios and quantities (concentrations). In the first experiment, we found a strong positive effect of the phosphorus content of food on growth of Daphnia, both in their early and late juvenile development. Variation in the relationship between the P-content of animals and their growth rate reflected interspecific differences in nutrient requirements. Although growth rates typically decreased as development neared maturation, this did not affect these species-specific couplings between growth rate and Daphnia P-content. In the second experiment, we examined the effects of food quality on Daphnia growth at different levels of food quantity. With the same decrease in P-content of food, species with higher estimated P-content at zero growth showed a larger increase in threshold food concentrations (i.e. food concentration sufficient to meet metabolic requirements but not growth). These results suggest that physiological processes such as maintenance and growth may in combination explain effects of food quality and quantity on consumers. Our study shows that differences in response to variation in food quality and quantity exist between species. As a consequence, species-specific effects of food quality on consumer growth will also determine how species deal with varying food levels, which has implications for resource-consumer interactions.