The possible role of toxigenic fungi in ecotoxicity of two contrasting oil-contaminated soils - A field study.

Long-term field experiments were performed to evaluate the phytotoxic properties of fungal metabolites in oil-contaminated soil and to assess the impact of contamination on the allelopathic activity of soil mycobiota. Two contrasting soils of Northwest Russia (sandy and loamy podzols) exposed to oil contamination underwent changes in abundance and allelopathic activities of soil fungi. Shifts within the microbial community caused by oil contamination affected not only oil-decomposition rates but also ecotoxicity of contaminated soil. There were significant differences in soil toxicity dynamics between sandy and loamy podzols. Four years after contamination, ecotoxicity of loamy podzol decreased, whereas sandy podzol remained highly toxic even nine years after contamination. The abundance and allelopathic activity of fungi is correlated with hydrocarbon degradation dynamics. The soil fungal community demonstrated high allelopathic activity which decreased over time in fertile loamy podzolic soil, whereas in poor sandy podzolic soil it remained high over the nine-year monitoring period. The results illustrate how oil contamination may influence allelopathic interactions in soil and demonstrate the advantage of using fungal metabolite toxicity test for testing of oil-contaminated soil samples.

The Influence of Changes in Magnetic Variations and Light-Dark Cycle on Life-History Traits of Daphnia magna.

Day-night cycle is the main zeitgeber (time giver) for biological circadian rhythms. Recently, it was suggested that natural diurnal geomagnetic variation may also be utilized by organisms for the synchronization of these rhythms. In this study, life-history traits in Daphnia magna were evaluated after short-term and multigenerational exposure to 16 h day/8 h night cycle, 32 h day/16 h night cycle, diurnal geomagnetic variation of 24 h, simulated magnetic variation of 48 h, and combinations of these conditions. With short-term exposure, the lighting mode substantially influenced the brood to brood period and the lifespan in daphnids. The brood to brood period, brood size, and body length of crustaceans similarly depended on the lighting mode during the multigenerational exposure. At the same time, an interaction of lighting mode and magnetic variations affected to a lesser extent brood to brood period, brood size, and newborn's body length. The influence of simulated diurnal variation on life-history traits in daphnids appeared distinctly as effects of synchronization between periods of lighting mode and magnetic variations during the multigenerational exposure. Newborn's body length significantly depended on the lighting regime when the periods of both studied zeitgebers were unsynchronized, or on the interaction of light regime with magnetic variations when the periods were synchronized. These results confirm the hypothesis that diurnal geomagnetic variation is an additional zeitgeber for biological circadian rhythms. Possible mechanisms for these observed effects are discussed. Bioelectromagnetics. © 2020 Bioelectromagnetics Society.

Transcriptome sequencing of a keystone aquatic herbivore yields insights on the temperature-dependent metabolism of essential lipids.

BACKGROUND: Nutritional quality of phytoplankton is a major determinant of the trophic transfer efficiency at the plant-herbivore interface in freshwater food webs. In particular, the phytoplankton's content of the essential polyunsaturated omega-3 fatty acid eicosapentaenoic acid (EPA) has been repeatedly shown to limit secondary production in the major zooplankton herbivore genus Daphnia. Despite extensive research efforts on the biological model organism Daphnia, and the availability of several Daphnia genomes, little is known regarding the molecular mechanisms underlying the limitations in Daphnia related to dietary EPA availability. RESULTS: We used RNA-seq to analyse the transcriptomic response of Daphnia magna which were fed with two different diets - each with or without supplementation of EPA - at two different temperature levels (15 and 20 °C). The transcripts were mapped to the D. magna genome assembly version 2.4, containing 26,646 translations. When D. magna fed on green alga, changing the temperature provoked a differential expression of 2001 transcripts, and in cyanobacteria-fed daphnia, 3385 transcripts were affected. The supplementation of EPA affected 1635 (on the green algal diet), or 175 transcripts (on the cyanobacterial diet), respectively. Combined effects for diet and temperature were also observed (669 for the green algal and 128 transcripts for the cyanobacterial diet). Searching for orthologous genes (COG-analysis) yielded a functional overview of the altered transcriptomes. Cross-matched transcript sets from both feed types were compiled to illuminate core responses to the factors temperature and EPA-supplementation. CONCLUSIONS: Our highly controlled eco-physiological experiments revealed an orchestrated response of genes involved in the transformation and signalling of essential fatty acids, including eicosanoid-signalling pathways with potential immune functions. We provide an overview of downstream-regulated genes, which contribute to enhance growth and reproductive output. We also identified numerous EPA-responsive candidate genes of yet unknown function, which constitute new targets for future studies on the molecular basis of EPA-dependent effects at the freshwater plant-herbivore interface.

Toxicological Evaluation and Quantification of Ingested Metal-Core Nanoplastic by Daphnia magna Through Fluorescence and Inductively Coupled Plasma-Mass Spectrometric Methods.

There are few studies on nanoplastic that propose quantification of the amount ingested combined with evaluation of the toxic effects on aquatic organisms. We propose 2 methods to quantify the amount of polystyrene nanoplastic (PSNP) ingested by Daphnia magna: fluorescence intensity, where a fluorescent monomer (F) is added to the PSNP and quantified through fluorescence light microscopy, and total aluminum quantification, where PSNP is synthesized with Al2 O3 metal-core nanoparticles and used for quantification of the nanoplastic ingested by the organism Daphnia magna using inductively coupled plasma-mass spectrometry. In addition, the PSNP was functionalized with palmitic acid to simulate the environmental conditions leading to biological and chemical transformations. Acute and chronic toxicity tests were performed with fluorescent PSNP (PSNP/F) and palmitic acid-functionalized PSNP/F (PSNP/F-PA). The ingestion quantified was higher by factors of 2.8 and 3.0 for PSNP/F-PA and 1.9 and 1.7 for PSNP/F applying the fluorescence intensity and total Al quantifying methods, respectively, when compared to PSNP. These results are consistent with the data obtained in the toxicity tests, which showed an approximately 3 times increase in the adverse effect of PSNP/F-PA on the mobility and reproduction of the organisms. Thus, the strong inhibition of D. magna reproduction caused by PSNP/F-PA in the chronic toxicity tests could be associated with a greater amount of this nanoplastic being ingested by the organisms. Environ Toxicol Chem 2019;38:2101-2110. © 2019 SETAC.

Water-soluble fraction of crude oil affects variability and has transgenerational effects in Daphnia magna.

The importance of interindividual variability in environmental responses has been little studied, although the available information suggests that, e.g., changes in environmental temperature may be associated with changes in variability. We studied, if exposure to water-soluble fraction (WSF) of crude oil can be associated with changes in interindividual variability in phenotype in Daphnia magna, which reproduces parthenogenetically. By using these clonal organisms, we could exclude the possibility that the observed changes were caused by genetic variability. The results show that the variability of oxygen consumption rate decreased in 48 h 30% WSF-exposed animals as compared to 10% WSF-exposed or control animals without a change in the mean of oxygen consumption rate. The clonal Daphnia magna could also be used to study transgenerational effects without genetic contribution, as the different generations are genetically identical. We observed that the oxygen consumption rates in F1 and F2 generations of unexposed and 10% WSF-exposed Daphnia had decreased from parental F0 generation and were also lower than in offspring of 30% WSF-exposed specimens. The studies did not aim at environmental realism but were designed to show the possibility of variability changes without changes in the mean value of a parameter, and transgenerational effects as a result of environmental contamination.

Assessment of Rhodopirellula rubra as a supplementary and nutritional food source to the microcrustacean Daphnia magna.

The daily use of the planctomycete Rhodopirellula rubra as an alternative or supplementary food source for Daphnia magna and its feasibility in the nutrition of transgenerational populations were studied. The life history parameters, fatty acids (saturated, mono- and polyunsaturated; SFAs, MUFAs and PUFAs), glycogen and protein contents of organisms during feeding assays and of the first generation were analysed. An increase in the yields of D. magna with the increase of the cell concentration of R. rubra was evident, but overall, bacteria supplied as the only food source was nutritionally insufficient as observed for all the parameters analysed. However, when R. rubra was added as supplement to the microalgae Raphidocelis subcapitata a significant improvement in the life history parameters was observed namely in the reproductive output and the somatic growth rate. The identified SFAs, MUFAs and PUFAs were the fatty acids more abundant in daphniids, and the feed regimens influenced daphniids fatty acid profiles. Additionally, the mixed diet resulted in a larger number and size of offspring in the different F1 broods as also observed with the results of F0 generation. The pink colouration present in D. magna body and eggs confirmed that bacteria were absorbed, the pigment(s) retained and passed on to the next generation. Our results showed that R. rubra can play an essential role in D. magna diet as a nutritional supplement showing potential biotechnological applications.

Stoichiometric responses to nano ZnO under warming are modified by thermal evolution in Daphnia magna.

Effects of stressors on body stoichiometry are important as these may cascade through food webs. Contamination and global warming are two key anthropogenic stressors, yet their effects on body stoichiometry have been rarely tested. Further, while thermal evolution may increase the ability to deal with warming, it is unknown how thermal evolution modifies the effect of contaminants under warming. Using resurrection ecology, we studied two Daphnia magna subpopulations (old/recent) of which the recent subpopulation evolved a higher heat tolerance. We exposed both subpopulations to a sublethal concentration of nano zinc oxide (nZnO) and 4 °C warming and quantified their effects on body stoichiometry: carbon (C), nitrogen (N), phosphorus (P) contents and their ratios (C:N, C:P, N:P). In the old subpopulation, nZnO only marginally decreased the C content and had no effect on N and P contents and their ratios. In contrast, in the recent subpopulation nZnO strongly increased the body P content (+51%) and reduced the C:P (-34%) and N:P (-34%) ratios at 24 °C but not at 20 °C. Moreover, these stoichiometric changes were not explained by changes of corresponding macromolecules as assumed by theory. Our results indicate that the stoichiometric responses to nZnO in Daphnia are temperature-dependent and modified by rapid evolution. The observed changes in body stoichiometry may affect the food quality of this important prey and have the potential to cascade through food webs and shape nutrients cycling.

Life history traits and genotoxic effects on Daphnia magna exposed to waterborne uranium and to a uranium mine effluent - A transgenerational study.

Assessing the impact of uranium mining industry on aquatic ecosystems near mining areas is critical to ensure the long-term health and sustainability of ecosystem services. As so, a transgenerational study with Daphnia magna has been conducted to perceive to what extent intermittent discharges of uranium mine effluents into watercourses may impact the DNA integrity and life history traits of cladocerans. Organisms were exposed for 48 h to a 2% dilution of an uranium mine effluent (UME) and to a corresponding dose of waterborne uranium (WU) that, according to our preliminary studies, induces significant DNA damage in daphnids. After exposure, organisms were transferred to clean medium, where three successive generations were monitored for genotoxicity and other effects at the individual and population level. Despite some differences between WU and UME data, our results revealed that the negative impacts of the short-term exposure gradually disappeared after placing the organisms in clean medium. These results suggest that, under intermittent stress, daphnids are able to recover, since after the 3rd brood release, DNA damage (measured as DNA strand breaks) is no longer observed and has no significant impact on the detectable life traits of offspring. Although our results indicate that populations of D. magna are not affected by intermittent and highly diluted discharges from uranium mining, aquatic systems under this kind of pressure should not be seen as hazardous-free. Future studies in this field are recommended and these should consider radionuclides in the water column, their accumulation in the sediments and also multiple life stages.

Impact of dispersant on early life stages of the water flea Daphnia magna and the eastern oyster Crassostrea virginica.

In response to the 2010 Deepwater Horizon oil spill, over 1 million gallons of dispersant were applied in Gulf of Mexico offshore waters; Corexit 9500 was the most applied dispersant. The impact on organisms in nearshore and freshwaters has received little scrutiny. Acute 48 h toxicity of Corexit 9500 and a new hyperbranched polyethylenimine (HPEI) dispersant-like compound were evaluated for the freshwater indicator organism, Daphnia magna and for larval and early spat stages of the Eastern oyster, Crassostrea virginica. For D. magna, Corexit 9500 demonstrated toxicity (EC50 of 0.14 [0.13, 0.15] ppm) similar to the 10-kDa HPEI (EC50 of 0.16 [0.12, 0.19] ppm). HPEI toxicity increased as a function of molecular weight (1.2 to 750 kDa). The 10 kDa size HPEI was further investigated because it dispersed crude oil with equal effectiveness as Corexit. For Corexit, 100% oyster mortality was detected for the ≤0.2-mm size classes and mortality >50% for the 0.3- and 0.7-mm size classes at the two greatest concentrations (25 and 50 ppm). HPEI (10 kDa) exhibited low mortality rates (<30%) for all concentrations for all oyster size classes except the 0.1-mm class. Although mortality rates for this size class were up to 60%, mortality was still less than the mortality caused by Corexit 9500. The low toxicity of HPEI polymers for C. virginica in comparison with Corexit 9500 suggests that HPEI polymers warrant further study.

Exchange of polar lipids from adults to neonates in Daphnia magna: Perturbations in sphingomyelin allocation by dietary lipids and environmental toxicants.

Because xenosensing nuclear receptors are also lipid sensors that regulate lipid allocation, we hypothesized that toxicant-induced modulation of HR96 activity would alter lipid profiles and the balance between adult survival and neonate production following exposure in Daphnia magna. Adult daphnids were exposed to unsaturated fatty acid- and toxicant- activators or inhibitors of HR96 and later starved to test whether chemical exposure altered allocation toward survival or reproduction. The HR96 activators, linoleic acid and atrazine, decreased reproduction as expected with concomitant changes in the expression of HR96 regulated genes such as magro. The HR96 inhibitors, docosahexaenoic acid (DHA) and triclosan, increased reproduction or neonate starvation survival, respectively. However, pre-exposure to triclosan increased in neonate survival at the expense of reproductive maturation. Lipidomic analysis revealed that sphingomyelins (SM) are predominantly found in neonates and therefore we propose are important in development. DHA and triclosan increased neonatal SM, consistent with HR96's regulation of Niemann-Pick genes. While DHA altered expression of magro, Niemann-Pick 1b, mannosidase, and other HR96-regulated genes as expected, triclosan primarily perturbed sphingomyelinase and mannosidase expression indicating different but potentially overlapping mechanisms for perturbing SM. Overall, SM appears to be a key lipid in Daphnia maturation and further support was provided by carmofur, which inhibits sphingomyelin/ceramide metabolism and in turn severely represses Daphnia maturation and initial brood production. In conclusion, toxicants can perturb lipid allocation and in turn impair development and reproduction.

Interactive effects of genotype and food quality on consumer growth rate and elemental content.

Consumer body stoichiometry is a key trait that links organismal physiology to population and ecosystem-level dynamics. However, as elemental composition has traditionally been considered to be constrained within a species, the ecological and evolutionary factors shaping consumer elemental composition have not been clearly resolved. To this end, we examined the causes and extent of variation in the body phosphorus (P) content and the expression of P-linked traits, mass specific growth rate (MSGR), and P use efficiency (PUE) of the keystone aquatic consumer Daphnia using lake surveys and common garden experiments. While daphnid body %P was relatively constrained in field assemblages sampled across an environmental P gradient, unique genotypes isolated from these lakes showed highly variable phenotypic responses when raised across dietary P gradients in the laboratory. Specifically, we observed substantial inter- and intra-specific variation and differences in daphnid responses within and among our study lakes. While variation in Daphnia body %P was mostly due to plastic phenotypic changes, we documented considerable genetic differences in daphnid MSGR and PUE, and relationships between MSGR and body P content were highly variable among genotypes. Overall, our study found that consumer responses to food quality may differ considerably among genotypes and that relationships between organismal life-history traits and body stoichiometry may be strongly influenced by genetic and environmental variation in natural assemblages.

Does infection tilt the scales? Disease effects on the mass balance of an invertebrate nutrient recycler.

While parasites are increasingly recognized as important components of ecosystems, we currently know little about how they alter ecosystem nutrient availability via host-mediated nutrient cycling. We examined whether infection alters the flow of nutrients through hosts and whether such effects depend upon host diet quality. To do so, we compared the mass specific nutrient (i.e., nitrogen and phosphorus) release rates, ingestion rates, and elemental composition of uninfected Daphnia to those infected with a bacterial parasite, P. ramosa. N and P release rates were increased by infection when Daphnia were fed P-poor diets, but we found no effect of infection on the nutrient release of individuals fed P-rich diets. Calculations based on the first law of thermodynamics indicated that infection should increase the nutrient release rates of Daphnia by decreasing nutrient accumulation rates in host tissues. Although we found reduced nutrient accumulation rates in infected Daphnia fed all diets, this reduction did not increase the nutrient release rates of Daphnia fed the P-rich diet because infected Daphnia fed this diet ingested nutrients more slowly than uninfected hosts. Our results thus indicate that parasites can significantly alter the nutrient use of animal consumers, which could affect the availability of nutrients in heavily parasitized environments.

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.

Dynamics of a producer-grazer model incorporating the effects of excess food nutrient content on grazer's growth.

Modeling under the framework of ecological stoichiometric allows the investigation of the effects of food quality on food web population dynamics. Recent discoveries in ecological stoichiometry suggest that grazer dynamics are affected by insufficient food nutrient content (low phosphorus (P)/carbon (C) ratio) as well as excess food nutrient content (high P:C). This phenomenon is known as the "stoichiometric knife edge." While previous models have captured this phenomenon, they do not explicitly track P in the producer or in the media that supports the producer, which brings questions to the validity of their predictions. Here, we extend a Lotka-Volterra-type stoichiometric model by mechanistically deriving and tracking P in the producer and free P in the environment in order to investigate the growth response of Daphnia to algae of varying P:C ratios. Bifurcation analysis and numerical simulations of the full model, that explicitly tracks phosphorus, lead to quantitative different predictions than previous models that neglect to track free nutrients. The full model shows that the fate of the grazer population can be very sensitive to excess nutrient concentrations. Dynamical free nutrient pool seems to induce extreme grazer population density changes when total nutrient is in an intermediate range.

A millennial-scale chronicle of evolutionary responses to cultural eutrophication in Daphnia.

For an accurate assessment of the anthropogenic impacts on evolutionary change in natural populations, we need long-term environmental, genetic and phenotypic data that predate human disturbances. Analysis of c. 1600 years of history chronicled in the sediments of South Center Lake, Minnesota, USA, revealed major environmental changes beginning c. 120 years ago coinciding with the initiation of industrialised agriculture in the catchment area. Population genetic structure, analysed using DNA from dormant eggs of the keystone aquatic herbivore, Daphnia pulicaria, suggested no change for c. 1500 years prior to striking shifts associated with anthropogenic environmental alterations. Furthermore, phenotypic assays on the oldest resurrected metazoan genotypes (potentially as old as c. 700 years) indicate significant shifts in phosphorus utilisation rates compared to younger genotypes. Younger genotypes show steeper reaction norms with high growth under high phosphorus (P), and low growth under low P, while 'ancient' genotypes show flat reaction norms, yet higher growth efficiency under low P. Using this resurrection ecology approach, environmental, genetic and phenotypic data spanning pre- and post-industrialised agricultural eras clearly reveal the evolutionary consequences of anthropogenic environmental change.

Transmission of DNA damage and increasing reprotoxic effects over two generations of Daphnia magna exposed to uranium.

This study aimed to examine the mechanisms involved in the transgenerational increase in Daphnia magna sensitivity to waterborne depleted uranium (DU) under controlled laboratory conditions. Daphnids were exposed to concentrations ranging from 2 to 50 µg L(-1) over two successive generations. Genotoxic effects were assessed using random amplified polymorphic DNA and real time PCR (RAPD-PCR). Effects on life history (survival, fecundity and somatic growth) were monitored from hatching to release of brood 5. Different exposure regimes were tested to investigate the specific sensitivity of various life stages to DU. When daphnids were exposed continuously or from hatching to deposition of brood 5, results demonstrated that DNA damage accumulated in females and were transmitted to offspring in parallel with an increase in severity of effects on life history across generations. When daphnids were exposed during the embryo stage only, DU exposure induced transient DNA damage which was repaired after neonates were returned to a clean medium. Effects on life history remained visible after hatching and did not significantly increase in severity across generations. The present results suggest that DNA damage might be an early indicator of future effects on life history.

Strong interactions between stoichiometric constraints and algal defenses: evidence from population dynamics of Daphnia and algae in phosphorus-limited microcosms.

The dynamic interactions among nutrients, algae and grazers were tested in a 2 × 3 factorial microcosm experiment that manipulated grazers (Daphnia present or absent) and algal composition (single species cultures and mixtures of an undefended and a digestion-resistant green alga). The experiment was run for 25 days in 10-L carboys under mesotrophic conditions that quickly led to strong phosphorus limitation of algal growth (TP is approximately equal to 0.5 µM, N:P 40:1). Four-day Daphnia juvenile growth assays tested for Daphnia P-limitation and nutrient-dependent or grazer-induced algal defenses. The maximal algal growth rate of undefended Ankistrodesmus (mean ± SE for three replicate microcosms; 0.92 ± 0.02 day(-1)) was higher than for defended Oocystis (0.62 ± 0.03 day(-1)), but by day 6, algal growth was strongly P-limited in all six treatments (molar C:P ratio >900). The P-deficient algae were poor quality resources in all three algal treatments. However, Daphnia population growth, reproduction, and survival were much lower in the digestion-resistant treatment even though growth assays provided evidence for Daphnia P-limitation in only the undefended and mixed treatments. Growth assays provided little or no support for simple threshold element ratio (TER) models that fail to consider algae defenses that result in viable gut passage. Our results show that strong P-limitation of algal growth enhances the defenses of a digestion-resistant alga, favoring high abundance of well-defended algae and energy limitation of zooplankton growth.

Larvicidal activity of ajowan ( Trachyspermum ammi ) and Peru balsam ( Myroxylon pereira ) oils and blends of their constituents against mosquito, Aedes aegypti , acute toxicity on water flea, Daphnia magna , and aqueous residue.

This study evaluated the larvicidal activity of 20 plant essential oils and components from ajowan ( Trachyspermum ammi ) and Peru balsam ( Myroxylon pereira ) oils against the mosquito, Aedes aegypti . Of the 20 plant essential oils, ajowan and Peru balsam oils at 0.1 mg/mL exhibited 100 and 97.5% larval mortality, respectively. At this same concentration, the individual constituents, (+)-camphene, benzoic acid, thymol, carvacrol, benzyl benzonate, and benzyl trans-cinnamate, caused 100% mortality. The toxicity of blends of constituents identified in two active oils indicated that thymol and benzyl benzoate were major contributors to the larvicidal activity of the artificial blend. This study also tested the acute toxicity of these two active oils and their major constituents against the water flea, Daphnia magna . Peru balsam oil and benzyl trans-cinnamate were the most toxic to D. magna. Two days after the treatment, residues of ajowan and Peru balsalm oils in water were 36.2 and 85.1%, respectively. Less than 50% of benzyl trans-cinnamate and thymol were detected in the water at 2 days after treatment. The results show that the essential oils of ajowan and Peru balsam and some of their constituents have potential as botanical insecticides against Ae. aegypti mosquito larvae.

Critical evaluation and further development of methods for testing ecotoxicity at multiple pH using Daphnia magna and Pseudokirchneriella subcapitata.

To meet the requirements of risk assessment legislature regarding the ecotoxicity of ionizing compounds, the present study attempts to establish easy, robust methods for testing ecotoxicity at various pH levels. An overview is given of the buffering methods found in the literature. This is supplemented by a series of experiments where toxicity and ability to stabilize pH of seven common buffering compounds was tested on Daphnia magna and Pseudokirchneriella subcapitata. We consider a buffer applicable at a given concentration if the pH drift is below 0.2 pH units, and if there are no toxic effects. Twenty-four- and 48-h acute toxicity tests with D. magna were carried on a series of organic buffers with pH monitoring. Based on the experimental results it is possible to give recommendations for buffer concentrations for use in toxicity testing with D. magna at pH levels in the range of pH 6.0-7.8 for 48 h exposure, and pH 6.0-9.5 for 24 h exposure. Forty-eight- and 72-h growth inhibition tests with P. subcapitata were carried out, and recommendations for buffer concentrations at pH 7.5 and 8.0 are made for both 48 and 72 h of exposure.

The potential of dietary polyunsaturated fatty acids to modulate eicosanoid synthesis and reproduction in Daphnia magna: a gene expression approach.

Nutritional ecology of the aquatic model genus Daphnia has received much attention in past years in particular with regard to dietary polyunsaturated fatty acids (PUFAs) which are crucial for growth and reproduction. Besides their significant role as membrane components, C20 PUFAs serve as precursors for eicosanoids, hormone-like mediators of reproduction, immunity and ion transport physiology. In the present study we investigate transcriptomic changes in Daphnia magna in response to different algal food organisms substantially differing in their PUFA composition using quantitative real-time PCR and relate them to concomitantly documented life history data. The selection of target genes includes representatives that have previously been shown to be responsive to the eicosanoid biosynthesis inhibitor ibuprofen. The beneficial effect of C20 PUFA-rich food on reproduction and population growth rates was accompanied by an increased vitellogenin (DmagVtg1) gene expression in D. magna. Additionally, genes involved in eicosanoid signaling were particularly influenced by dietary C20 PUFA availability. For example, the cyclooxygenase gene (Cox), coding for a central enzyme in the eicosanoid pathway, was highly responsive to the food treatments. Our results suggest that dietary PUFAs are fundamental in D. magna physiology as substrate for eicosanoid synthesis and that these eicosanoids are important for D. magna reproduction.