Alterations of physiological energetics, growth and reproduction of Daphnia magna under toxicant stress

The study investigates the relationship between changes in physiological energetics of organisms and alterations of growth, development and reproduction of Daphnia magna. Groups of primiparous daphnids were subjected to 8-day exposures to the heavy metals cadmium and copper or to the cationic surfactant, cetyltrimethylammonium bromide (CTAB). Energetic alterations were estimated from the measurement of oxygen consumption and feeding activity which was performed during the last 3 days of the exposure period and from the calculation of simplified carbon balances. The physiological effects were compared to effects on organismal growth and reproduction as obtained from 17-day exposure experiments. Toxicant exposure reduced weight and body length of daphnids indicating an impaired growth rate, but effects on total metabolic costs measured as weight-specific oxygen consumption could not be detected. Net carbon gain of individuals decreased in a concentration-dependent way for the tested chemicals reflecting effects on biomass of daphnids. In the case of cadmium and copper, reproduction ( summation operatormx: number of offspring per female of age x born during the time interval x-1 to x, summarised over the entire exposure period) and the estimate for the intrinsic rate of natural increase, derived from the 17-day exposure-experiment, were affected at concentrations comparable to the effect levels as observed for growth. In the case of copper, the concentrations affecting growth and reproduction were close to the 17-day LC(50) value. CTAB caused a reduction in body length of primiparous daphnids whereas a decrease in the reproductive performance was not apparent. In conclusion, the chemicals did not change metabolic costs of exposed daphnids as it would be expected as a consequence of resistance or repair mechanisms, however, they induced alterations of SFG, growth, reproduction and intrinsic rate of natural increase. These alterations were chemical-specific. The fact that toxicant-related effects on growth and reproduction could not be linked to an elevated metabolic rate of daphnids may indicate that demand side effects occurred early during exposure - before the start of respirometric measurements - or that effects on growth were caused by an altered energy uptake. The results illustrate the importance of trade-off processes in regulating the distribution of energy among growth and reproduction of daphnids.     

Temperature influences on the accumulation and elimination of mercury in a freshwater cladoceran, Daphnia magna

Temperature is an important environmental variable affecting the physiology and metabolism of aquatic invertebrates and can potentially affect the rate and pathway of the uptake and elimination of contaminants. In this study, we studied the effects of the ambient temperature on the uptake (from water and food) and elimination of inorganic mercury [Hg(II)] and methymercury (MeHg) in a freshwater cladoceran, Daphnia magna, in the laboratory. At different temperatures (i.e., 14, 19 and 24 degrees C), the assimilation efficiencies of both Hg species from ingested alga were not significantly different while the dissolved uptake rates were 32 and 73% lower at 14 degrees C than at 24 degrees C for Hg(II) and MeHg, respectively. The reduced uptake rates of Hg(II) at 14 degrees C could be adequately explained by the reduced filtration rate (40% reduction), but for MeHg, the reduced filtration rate could not completely explain reduction in MeHg uptake rate. Despite the elimination rates of both Hg compounds not being affected by the temperature, the relative importance of different routes of Hg loss (i.e., excretion, egestion, molting and reproduction) was significantly different at various temperatures. The relative importance of excretion to MeHg loss increased from 24 to 14 degrees C (i.e., 52 to 85% of Hg loss) while that of reproduction to MeHg loss decreased from 43 to 11% simultaneously. Using a kinetic model, we showed that the fraction of MeHg accumulated in the daphnids coming from the dietary phase was higher at lower temperature, while there was increased accumulation of both Hg(II) and MeHg in the daphnids at higher temperatures. Nevertheless, the trophic transfer potentials for both Hg species in this algae-daphnids food chain were not influenced by the temperature.     

Effects of N-heterocyclic polyaromatic hydrocarbons on survival, reproduction, and biochemical parameters in Daphnia magna

N-heterocyclic polycyclic aromatic hydrocarbons (N-PAHs) belong among newly identified classes of environmental pollutants with relatively high toxic potential. N-PAHs have been detected in air, soil, marine environments, and freshwater sediments. The N-PAHs are present at lower concentrations than their nonsubstituted analogues but their greater solubility would lead to greater bioavailibity and potential for toxic effects. Here we present results of acute and chronic toxicity in traditional aquatic invertebrate ecotoxicological model (Daphnia magna) along with assessment of biochemical responses. Studied biomarkers in D. magna exposed to N-heterocyclic derivatives included glutathione levels and activities of detoxication and antioxidative enzymes glutathione S-transferase and glutathione peroxidase. Phenanthrene and 1,10-phenathroline were the most toxic of all tested compounds (EC50 < 6 microM after 48 h exposure) and all tested N-PAHs suppressed reproduction of Daphnia magna. The data suggest that N-PAHs can induce oxidative stress in D. magna. The significant decline of glutathione content was found in animals treated with acridine, 1,10-phenanthroline, benzo(h)quinoline, phenantridine, and phenazine. Significant decrease of GPx activities relative to controls was found for all tested compounds except of phenanthrene and phenazine. Activities of GST increased after exposure to phenanthridine, phenazine, and benzo(h)quinoline, and declined in D. magna treated with phenanthrene (significant at one concentration) or anthracene (not significant). Our results confirmed significant acute as well as chronic toxicities of N-PAHs as well as potential of biochemical parameters to be used as early warning signals of toxicity in Daphnia magna.     

Chronic toxicity of Great Lakes sediments to Daphnia magna: elutriate effects on survival, reproduction and population growth

Seventeen Great Lakes sediments ranging in degree of expected toxicity were evaluated using a 21 day sediment elutriate bioassay with Daphnia magna. Sediments differed in their effects on survival, age at first reproduction, the number of broods produced and the total number of young produced per adult. Sediments producing low survivorship (50–60%) also had negative effects on reproduction. However, both positive and negative effects on reproduction were found among sediments producing high survivorship. To integrate all test end-points, a stochastic matrix population model was constructed and parameterized with survival and reproduction data from each sediment. By including estimates of variability in vital rates, the model output provided quantitative estimates of uncertainty in projected population size. Sediment effects on survival and reproduction translated into large differences in projected population growth; mean estimates of projected population size at day 28 of the simulations ranged over two orders of magnitude among the 17 sediments. Reproductive timing (e.g. age at first reproduction), followed by fecundity and survivorship, had the largest effect on population growth. Results of this study also indicate that the presence of suspended sediment in elutriates may confound toxicity evaluations using cladocerans. The concentration of total suspended solids was negatively correlated with age at first reproduction and positively correlated with measures of fecundity and population growth. In order to realize the potential benefits of chronic testing we must develop ecologically relevant ways of interpreting sediment bioassay results and expressing the uncertainty associated with our estimates of ecological risk.     

Effect of cadmium on the reproduction strategy of Daphnia magna

Survival, number of neonates per female, first day of reproduction and neonate size were investigated in Daphnia magna, exposed to cadmium concentrations ranging from 0 to 50.0 ppb. A 25-day LC₅₀ of about 10.0 ppb was found. The onset of reproduction was delayed at concentrations of cadmium above 5.0 ppb, but was not affected at lower levels. Cadmium concentrations of 0.5, 1.0 and 5.0 ppb were shown to increase the average number of neonates per female significantly. The size of the neonates, however, decreased significantly with increasing cadmium concentrations, thereby probably offsetting their survival probability. The reproduction strategy of D. magna seems to be changed by low concentrations of cadmium. Compared with unstressed animals cadmium-stressed animals produce larger broods with smaller neonates, but at higher cadmium concentrations brood size and body size both decline. Whether the former observation might be considered an adaptation of the stressed animal, or whether ‘hormesis’, i.e., the unspecific effect on the growth control mechanism, might be responsible, is discussed.     

The interactive effects of a cadmium stress,a selenium deficiency and water temperature on the survival and reproduction of Daphnia magna Straus

The interactive effects of a Se deficiency and a Cd stress on Daphnia magna were evaluated at 20 and 25°C over a 25-day exposure. The effects of a Se deficiency and of a Cd stress differed at the two temperatures. A Se deficiency had no observable direct effect on daphnids maintained at 20°C but resulted in massive abortions of eggs in animals maintained at 25°C. A Cd stress caused an increase in mortality, a decrease in egg production and a decrease in the number of broods/female at 20°C but had no effect on survival at 25°C. Its effect on reproduction at the higher temperature was an increase in the abortion rate and a decrease in the number of broods/female. Based on total young/female, Se-enriched animals were more sensitive to a Cd stress at 25°C, and Se-deficient animals were more sensitive at 20°C. This was because Se-deprived animals were more sensitive to 1.5 μg Cd/l than control animals at 20°C but not at 25°C. At an exposure of 3.0 μg Cd/l, Se-deprived animals were more sensitive than Se-enriched animals at both temperatures.     

Toxicity of soybean rust fungicides to freshwater algae and Daphnia magna

Soybeans are intensively grown over large swaths of land in the Midwestern US. Introduction of the pathogenic fungus responsible for Soybean Rust (Phakopsora pachyrhizi) will likely result in a significant increase in the environmental load of strobilurin and conazole fungicides. We determined the toxicity of six such fungicides to the unicellular algae Pseudokirchneriella subcapitata and the aquatic invertebrate, Daphnia magna. We found that levels of concern of some fungicides were lower than annual average runoff concentrations predicted for Indiana. Our results suggest that pyraclostrobin and propiconazole, and to a lesser extent tebuconazole, may cause impacts to algae and daphnids in areas where soybeans are intensively grown. More studies are needed to describe the ecological effects of sublethal exposures to these fungicides, as well as monitoring environmental concentrations in watersheds where these fungicides are applied to soybeans.     

Behavioural response of the cladoceran Daphnia magna STRAUS to sublethal Copper stress--validation by image analysis

In this study changes in the locomotory behaviour of the freshwater cladoceran Daphnia magna STRAUS were used as sublethal indicators of toxic copper (Cu) stress. The behavioural responses were determined by a real time image analysis, using a video camera and a Pentium-PC equipped with a standard low cost frame grabber. For a sequence of 50 images per group, where 10 daphnids were moving simultaneously, the trajectories have been reconstructed in binary image sequences. As biological endpoints, we defined the average swimming velocity and the average duration of swimming activity and inactivity. The behavioural responses of the daphnids were analysed under normal conditions (without Cu stress) and after application of sublethal Cu stress of following concentrations (in microg l(-1) (ppb)): C1=1, C2=5, C3=10, C4=20, C5=30. The test organisms were exposed to the Cu concentration for 24 h under static conditions. Already after 9 h of Cu-exposure a significant (*P<0.05) decrease of the average swimming velocity could be observed at the group of the highest Cu concentration (30 ppb). After 13 h of Cu-contamination the swimming velocity was significantly (*P<0.05) reduced at group of 20 ppb Cu-treatment and after 14 h a significant (*P<0.05) decrease of the average swimming velocity could be measured at the group of 10 ppb Cu-treatment. No significant decrease of the swimming velocity could be observed in the 1 and 5 ppb Cu-treatment.     

Optimization of differential display polymerase chain reaction as a bioindicator for the cladoceran Daphnia magna

Research on toxicant-responsive genes is providing new and important bioindicators for environmental biologists. Identifying genes whose expression is modulated by toxicant exposure provides important clues into the mechanisms underlying toxicity. In addition, toxicant-responsive genes can be developed as molecular end points that are likely to be sensitive tools for environmental assessment. Differential display polymerase chain reaction (ddPCR) is a useful approach for screening and analyzing the expression of genes. A ddPCR protocol was optimized to investigate gene expression in the cladoceran Daphnia magna. The modified protocol requires submicrogram quantities of total RNA (from <10 animals) and utilizes a sensitive fluorescent tagging system. By reverse-transcribing total RNA with arbitrary 18-nucleotide primers and PCR-amplifying the cDNA using the same arbitrary primers under low-stringency conditions, reproducible and consistent ddPCR profiles were generated. Minimal variability was introduced by reaction differences or biological variability. A trial stress (starvation) was found to generate modest differences in the ddPCR profiles. This technique promises to significantly advance knowledge regarding gene expression during toxicant insult. Furthermore, this represents the first step in the development of a novel gene fingerprinting technique that can be applied to any compound and organism of interest.     

Effects of chronic internal alpha irradiation on physiology, growth and reproductive success of Daphnia magna

Daphnids were chronically exposed to waterborne Am-241, an alpha-emitting radionuclide, ranging in concentration from 0.4 to 40 Bq ml(-1). Am-241 amounts were monitored in the medium, daphnid tissues and cuticles. Corresponding average dose rates of 0.02, 0.11 and 0.99 mGy h(-1) were calculated for whole organisms with internal alpha-radiation contributing 99% of total dose rates. Effects of internal alpha irradiation on respiration and ingestion rates, adult, egg and neonate individual dry masses, fecundity and larval resistance to starvation were examined in 23-day experiments. Daphnids showed increased respiratory demand after 23 days at the highest dose rate, suggesting increased metabolic cost of maintenance due to coping with alpha radiological stress. Although no effect was detected on ingestion rates between contaminated and control daphnids, exposure to dose rates of 0.11 mGy h(-1) or higher, resulted in a significant 15% reduction in body mass. Fecundity remained unchanged over the 23-day period, but individual masses of eggs and neonates were significantly smaller compared to the control. This suggested that increased metabolic expenditure in chronically alpha-radiated daphnids came at the expense of their energy investment per offspring. As a consequence, neonates showed significantly reduced resistance to starvation at every dose rate compared to the control. Our observations are discussed in comparison with literature results reported for cadmium, a chemical toxicant which affects feeding activity and strongly reduces individual energy uptake.     

Dynamic multipathway modeling of Cd bioaccumulation in Daphnia magna using waterborne and dietborne exposures

We tested the predictive ability of the dynamic multipathway bioaccumulation model (DYMBAM) to characterize Cd accumulation in Daphnia magna, a species commonly used in toxicity tests and because of its sensitivity, particularly to metals, a species that is relied upon in ecological risk assessments. We conducted chronic exposure experiments in which D. magna were exposed to either dietborne Cd alone or to both dietborne and waterborne Cd. In the food-only treatments, the algae Chlamydomonas reinhardtii or Pseudokirchneriella subcapitata were pre-exposed to free Cd ion concentrations, [Cd(2+)], from 0.001 to 100nM (0.001-11microgL(-1)) then, on a daily feeding renewal basis, fed to D. magna over 21 days. In the water plus food treatment, D. magna were exposed for 21 days to the same range of [Cd(2+)] and fed with the same algal species that had been exposed to Cd at various concentrations. In the algal exposure media, Cd concentrations in algae were directly related to those in water and were characterized by a linear regression model using the log transformed concentration of the WHAM predicted Cd(2+) concentration. The DYMBAM was used with estimated values of the model constants for ingestion rate (0.08-0.34gg(-1)day(-1)) and growth rate (0.085-0.131day(-1)) based on our experimental data and with literature values for rate constants of Cd influx and efflux as well as Cd assimilation efficiency. Measured Cd concentrations in D. magna agreed with model predictions within a factor of 3. Using the model, we predict that food is an important contributor of Cd burden to D. magna, particularly at lower Cd exposure concentrations over an environmentally realistic gradient of free Cd in water. However, this cladoceran also takes up Cd from water and this exposure route becomes increasingly important at very high concentrations of free Cd (>10nM or 1.1microgL(-1)). Nevertheless, Cd produced lethal effects in D. magna that were exposed to this metal in water and diet, but exposure to Cd in food only did not result in toxic effects (as measured by survival and reproduction).     

Effects of microcystin-free and microcystin-containing strains of the cyanobacterium Microcystis aeruginosa on growth of the grazer Daphnia magna

Harmful effects of the common bloom-forming cyanobacterium Microcystis aeruginosa on the grazer Daphnia have been explained from morphological features, nutritional insufficiency, and the production of toxins called microcystins. The effects of four M. aeruginosa strains, including one free of microcystins, on the growth of Daphnia magna were examined. Population growth was measured in Daphnia fed each M. aeruginosa strain alone and fed mixtures of each strain with the green alga Scenedesmus obliquus to avoid nutritional deficiency. The presence of Microcystis in the food resulted in decreased Daphnia population growth rates. A significant correlation (P < 0.001) was found between the amount of microcystins (MC) in the food and the population growth rate, but growth inhibition in Daphnia fed with the MC-free strain could not be explained by the toxicity of the microcystins. Inasmuch as there was greater reduction in growth of the Daphnia fed mixtures of the MC-free strain with Scenedesmus than of the Daphnia fed only the same amount of Scenedesmus as in the mixtures, other factors associated with this Microcystis strain must have inhibited growth. However, morphology and nutritional insufficiency could be excluded as causal factors. The strong relationship between Daphnia feeding activity and population growth suggests that feeding inhibition may have caused the observed detrimental effects on the animals.     

Quantitative structure-activity relationships and mixture-toxicity studies of alcohols and chlorohydrocarbons: reproducibility of effects on growth and reproduction of Daphnia magna

In subacute toxicity experiments for 10 nonreactive organic chemicals, such as alcohols and chlorobenzenes, the effects on growth and reproduction of Daphnia magna were studied. No-observed-effect concentrations (NOEC) for growth and reproduction were found to be approximately equal. For all compounds tested, these values and the median effective concentrations (EC₅₀) on reproduction correlated very well with hydrophobicity (octanol-water partition coefficient, P_oct.) as shown by a quantitative structure-activity relationship (QSAR) study.From one of the QSAR equations, EC₅₀ values for 15 other compounds were predicted, after which the toxicity of mixtures of 10 and 25 compounds was tested. The toxicity was found to be predictable on the basis of concentration addition. Comparing the results of this study with those from earlier experiments, it may be concluded that subacute toxicity data for D. magna are very reproducible and are not very dependent upon the physical condition of the daphnids used.     

Effects of far-infrared ray on reproduction, growth, behaviour and some physiological parameters in mice

The effects of chronic exposure to far-infrared ray (FIR) on reproduction, growth, behaviour, survival time and some related parameters were examined in SHN mice. The reproductive parameters differed slightly between the females on the normal racks and those on the FIR racks, which emitted FIR from the ceiling. The age and body weight on the day of vaginal opening was lower in the experimental mice born and maintained on the FIR rack than in the control on the normal rack. In both sexes, the levels of urinary components in the experimental group was significantly higher than the control at 6-7 months of age. Spontaneous motor activity of females during the light and dark phases were higher and lower, respectively, in the experimental group than the control. The survival rate was significantly higher in the experimental group than the control. These findings suggest that FIR has 'normalization effects' on the organisms.     

Accumulation of paralytic shellfish toxins (PST) from the cyanobacterium Aphanizomenon issatschenkoi by the cladoceran Daphnia magna.

In order to access the effects of Paralytic Shellfish Toxins (PST) in freshwater environment, the accumulation of PST produced by the cyanobacteria Aphanizomenon issatschenkoi in juvenile Daphnia magna was investigated. D. magna was exposed to A. issatschenkoi cells (1.2 x 10(6) cells ml(-1)) for 6, 8, 12, 24 and 30 h and also to lyophilised material (1 mg ml(-1)) for 24h. Survival and somatic growth of the juvenile D. magna was investigated, as was the activity of the biotransformation enzyme system glutathione-S-transferases (GSTs). Between 643+/-65.35 and 1170+/-51.72 pmol PST ml(-1) were detected by HPLC-FLD in D. magna culture medium containing cells and 2745+/-64.61 pmol PST toxin ml(-1), in the medium containing lyophilised material. PST were detected in D. magna tissues in cells exposure (between 6.51 x 10(-2)+/-1.37 x 10(-2) and 3.78 x 10(-1)+/-1.15 x 10(-2)pmol PST animal(-1)). In D. magna exposed to lyophilised material the mean (+/-SD) PST concentration was found to be 6.96 x 10(-3) (+/-3.84 x 10(-3)) pmol PST animal(-1). Following exposure to 1.2 x 10(6) cells ml(-1)A. issatschenkoi fresh cells growth and survival of D. magna were reduced. D. magna exposed to the two A. issatschenkoi treatments (fresh cells and lyophilised material), showed a reduction in activity of the cytosolic glutathione-S-transferases (cGSTs). The results of this study indicate that D. magna can accumulate PST toxins and that the cyanobacterium A. issatschenkoi affects both the fitness and growth potential of juvenile D. magna.     

Low exposure concentration effects of methoprene on endocrine-regulated processes in the crustacean Daphnia magna.

Methoprene is a growth-regulating insecticide that manifests its toxicity to target organisms by acting as a juvenile hormone agonist. Methoprene similarly may exert toxicity to crustaceans by mimicking or interfering with methyl farnesoate, a crustacean juvenoid. We hypothesized that methoprene interferes with endocrine-regulated processes in crustaceans by several mechanisms involving agonism or antagonism of juvenoid receptor complexes. In the present study, we evaluated this hypothesis, in part, by characterizing and comparing the concentration-response curves for methoprene and several endpoints related to development and reproduction of the crustacean Daphnia magna. Our results demonstrate that methoprene has multiple mechanisms of toxicity and low-exposure concentration effects. Methoprene reduced the growth rate of daphnids with evidence of only a single concentration-response line, having a threshold of 12.6 nM. Molt frequency was reduced by methoprene in a concentration-dependent manner, with a response curve corresponding to a 2-segmented line and thresholds at 4.2 and 0.21 nM. An endpoint related to reproductive maturation, the time of first brood deposition, was also affected by methoprene, with a clear concentration-dependent response and a NOEC of 32 nM. Methoprene reduced fecundity according to a 2-segmented line, with thresholds of 24 and < or =0.18 nM. These results demonstrate that methoprene elicits significant toxicity to endocrine-related processes in the 5-50 nM concentration range. Furthermore, molting and reproduction were impacted at significantly lower methoprene concentrations, with a distinct concentration response and a threshold of < or =0.2 nM. The different concentration-dependent response from that of methoprene could involve agonism or antagonism of various juvenoid receptor configurations.     

Differentiation of the toxicities of silver nanoparticles and silver ions to the Japanese medaka (Oryzias latipes) and the cladoceran Daphnia magna

Abstract

Silver nanoparticles (AgNPs) are increasingly used in various fields. However, little is known about the environmental effects of widespread use of products containing AgNPs. The objective of this study was to determine the ecotoxicity caused by AgNPs. The 48-h effective concentration 50 (EC₅₀) values for Daphnia magna of suspensions of 60 nm and 300 nm AgNPs were 1.0 (95% confidence interval [CI] = 0.1–1.3) and 1.4 (95% CI = 0.3–2.1) μg Ag/l, respectively. The 96 h LC₅₀ values for Oryzias latipes of 60 nm and 300 nm AgNP suspensions were 28 (95% CI = 23–34) and 67 (95% CI = 45–108) μg Ag/l, respectively. To show that toxicity is caused only by Ag⁺ and not by AgNPs, Ag⁺ was adsorbed onto the synthesized sorbents packed in a column and D. magna was exposed to the column-passed-300 nm AgNP suspensions. There was no acute toxicity with the AgNP suspensions not containing Ag⁺.