Connie J. Boese

Whole-body accumulation of copper predicts acute toxicity to an aquatic oligochaete (Lumbriculus variegatus) as pH and calcium are varied☆

We tested the hypothesis that whole-body accumulation of Cu at 50% mortality (i.e. the median lethal accumulation, LA50 value) in a freshwater oligochaete (Lumbriculus variegatus) is constant across a wide range of water quality, whereas the LC50 values of Cu(total) and the cupric ion (Cu(2+)) in solution are not. We exposed the worms in intermittent-flow, water-only chambers to a series of Cu concentrations at a variety of combinations of pH and water hardness (pH 6.5, 7.5 and 8.5 crossed with hardnesses of 0.5, 2, 4, 6 and 15 mEql(-1)) at 17-20 degrees C. In addition to monitoring mortality at 48 h, we determined whole-body Cu uptake in half of the replicate chambers at 6 h. LC50 values of Cu(total) and Cu(2+) increased as water hardness increased, as expected from traditional LC50 vs. hardness regressions. Moreover, LC50 values of Cu(total) remained approximately constant and LC50 values of Cu(2+) decreased considerably as pH increased, as expected from principles of cation competition and binding by inorganic ligands. However, LA50 values of Cu(body) remained approximately constant (0.17-0.34 micromol Cug(-1) dry wt.) in all pH x hardness combinations. Thus, consistent with the biotic-ligand model, Cu accumulation might be a constant predictor of acute mortality to L. variegatus whereas aqueous Cu concentrations are not.

Acute toxicity of copper and silver to Ceriodaphnia dubia in the presence of food

Food is added to exposure solutions in cladoceran chronic toxicity tests and sometimes in acute toxicity tests, but its effects on the bioavailability of toxicants have not been studied extensively. We compared the toxicity of waterborne Ag and Cu to Ceriodaphnia dubia in the presence or absence of food (a mixture of a yeast--Cerophyll--trout chow slurry and a green alga) in two series of acute toxicity tests. In the first series, we added food to Ag or Cu exposure solutions 0, 30, 60, or 120 min before transferring C. dubia into the solutions. In the second series, we exposed C. dubia to waterborne Ag or Cu for 30, 60, and 120 min in the absence of food. Adding food before transferring C. dubia into the exposure solutions greatly decreased the toxicity of Ag, but had less effect on the toxicity of Cu. In contrast, adding food after transferring C. dubia into the exposure solutions did not alter the toxicity of Ag considerably and did not alter the toxicity of Cu as much as the reverse sequence. Median effects concentrations (EC50s) for 30-, 60-, and 120-min unfed Ag tests were within the range of EC50s for 48-h unfed Ag tests, suggesting most uptake that contributes to acute Ag toxicity to C. dubia occurs within the first 30 to 60 min. However, uptake that contributes to acute Cu toxicity to C. dubia appears to occur over more than 2 h. Therefore, standard food decreases the toxicity of waterborne Ag and Cu to C. dubia, and the timing of adding food to exposure solutions is especially important in Ag tests.

Effects of dietborne copper and silver on reproduction by Ceriodaphnia dubia

Recent studies have indicated the potential for dietborne metals as an important exposure pathway for metal toxicity in freshwater organisms. We conducted a study in which freshwater cladocerans (Ceriodaphnia dubia) were fed green algae (either Pseudokirchneriella subcapitata or Chlorella vulgaris) that were grown in Ag- or Cu-contaminated media. In one series of toxicity tests patterned after the U.S. Environmental Protection Agencys three-brood C. dubia chronic toxicity test, we exposed C. dubia to waterborne Ag or Cu while feeding them normal amounts of uncontaminated yeast-Cerophyll-trout chow (YCT) slurry and either algae grown in standard media or algae grown in standard media supplemented with Ag or Cu (added as AgNO3 or CuSO4 x 5H2O). These parallel tests demonstrated that dietborne metal did not contribute to survival or reproduction effects beyond the effects caused by waterborne metal alone. We also conducted dietborne-only toxicity tests patterned after two other recently published experimental designs in which (1) C. dubia were fed only metal-contaminated algae for 4 h, transferred to fresh water, and fed uncontaminated algae and YCT slurry for the duration of the three-brood test or (2) C. dubia were fed standard amounts of metal-contaminated algae and uncontaminated YCT slurry for the entire three-brood test. In contrast to previous studies, we did not find consistent dietborne metal toxicity or standard concentration-response relationships in those two experiments. Instead, among-experiment variation in intracellular partitioning of metals in the algae fed to the C. dubia, among-laboratory differences in experimental procedures, selective feeding by C. dubia to avoid metal-contaminated algae, an interaction between reproductive status of the C. dubia and dietborne metal concentration, or a combination of these might help explain the apparently inconsistent results.

Dissolved fraction of standard laboratory cladoceran food alters toxicity of waterborne silver to Ceriodaphnia dubia

The biotic ligand model (BLM) for the acute toxicity of cationic metals to aquatic organisms incorporates the toxicity-modifying effects of dissolved organic matter (DOM), but the default parameterization (i.e., assuming 10% of DOM is humic acid) does not differentiate DOM from different sources. We exposed a cladoceran (Ceriodaphnia dubia) to Ag in the presence of DOM from filtered YCT (standard yeast–Cerophyll®–trout chow food recommended by the U.S. Environmental Protection Agency [EPA] for cladocerans), from the Suwannee River (GA, USA; relatively little anthropogenic input), and from the Desjardins Canal in Hamilton (ON, Canada; receives treated municipal wastewater effluent). In all three treatments, the dissolved organic carbon (DOC) concentration was 2 mg/L (the concentration following addition of YCT slurry at the U.S. EPA–recommended volume ratio). The average 48-h median effects concentration (EC50) ratios for dissolved Ag in the presence and absence of DOM [i.e., (EC50 with DOM)/(EC50 without DOM)] were as follows: Suwannee River, 1.6; Desjardins Canal, 2.2; and YCT filtrate, 26.8. Therefore, YCT filtrate provided much more protection against Ag toxicity than that provided by DOM from the surface waters. The major spectral characteristic that differentiated YCT filtrate from the other two types of DOM was a strong tryptophan peak in the excitation–emission matrix for YCT. These results have important implications for interpreting Ag toxicity tests in which organisms are fed YCT, and they suggest BLM-calculated toxicity predictions might be improved by incorporating specific chemical constituents or surrogate indices of DOM. Another component of the protective effect against Ag toxicity, however, might be that the dissolved fraction of YCT served as an energy and/or nutrient source for C. dubia.