Root length of aquatic plant, Lemna minor L., as an optimal toxicity endpoint for biomonitoring of mining effluents.

Lemna minor, a free-floating macrophyte, is used for biomonitoring of mine effluent quality under the Metal Mining Effluent Regulations (MMER) of the Environmental Effects Monitoring (EEM) program in Canada and is known to be sensitive to trace metals commonly discharged in mine effluents such as Ni. Environment Canada's standard toxicity testing protocol recommends frond count (FC) and dry weight (DW) as the 2 required toxicity endpoints-this is similar to other major protocols such as those by the US Environmental Protection Agency (USEPA) and the Organisation for Economic Co-operation and Development (OECD)-that both require frond growth or biomass endpoints. However, we suggest that similar to terrestrial plants, average root length (RL) of aquatic plants will be an optimal and relevant endpoint. As expected, results demonstrate that RL is the ideal endpoint based on the 3 criteria: accuracy (i.e., toxicological sensitivity to contaminant), precision (i.e., lowest variance), and ecological relevance (metal mining effluents). Roots are known to play a major role in nutrient uptake in conditions of low nutrient conditions-thus having ecological relevance to freshwater from mining regions. Root length was the most sensitive and precise endpoint in this study where water chemistry varied greatly (pH and varying concentrations of Ca, Mg, Na, K, dissolved organic carbon, and an anthropogenic organic contaminant, sodium isopropyl xanthates) to match mining effluent ranges. Although frond count was a close second, dry weight proved to be an unreliable endpoint. We conclude that toxicity testing for the floating macrophyte should require average RL measurement as a primary endpoint.

Effect of major cations (Ca2+, Mg2+, Na+, K+) and anions (SO4(2-), Cl- , NO3-) on Ni accumulation and toxicity in aquatic plant (Lemna minor L.): implications For Ni risk assessment.

The effect of major cation activity (Ca(2+) , Mg(2+) , Na(+) , K(+) ) on Ni toxicity, with dose expressed as exposure (total dissolved Ni concentration NiTot ) or free Ni ion activity (in solution Ni(2+) ), or as tissue residue (Ni concentration in plant tissue NiTiss ) to the aquatic plant Lemna minor L. was examined. In addition, Ni accumulation kinetics was explored to provide mechanistic insight into current approaches of toxicity modeling, such as the tissue residue approach and the biotic ligand model (BLM), and the implications for plant Ni risk assessment. Major cations did not inhibit Ni accumulation via competitive inhibition as expected by the BLM framework. For example, Ca(2+) and Mg(2+) (sulfate as counter-anion) had an anticompetitive effect on Ni accumulation, suggesting that Ca or Mg forms a ternary complex with Ni-biotic ligand. The counter-anion of the added Ca (sulfate, chloride, or nitrate) affected plant response (percentage of root growth inhibition) to Ni. Generally, sulfate and chloride influenced plant response while nitrate did not, even when compared within the same range of Ca(2+) , which suggests that the anion dominated the observed plant response. Overall, although an effect of major cations on Ni toxicity to L. minor L. was observed at a physiological level, Ni(2+) or NiTot alone modeled plant response, generally within a span of twofold, over a wide range of water chemistry. Thus, consideration of major cation competition for improving Ni toxicity predictions in risk assessment for aquatic plants may not be necessary.

Comparative study of the fate and mobility of metals discharged in mining and urban effluents using sequential extractions on suspended solids.

The fate, bioavailability and environmental impacts of metals discharged in municipal and mining wastewater discharge will depend to a large extent on chemical speciation and distribution. Previous studies on metal bioaccumulation have shown that total metal concentrations are not a good predictor of bioavailability in the dispersion plumes of municipal effluents. The objective of this study was to determine the solid phase speciation of metals in surface waters receiving urban and mining effluents in order to assess their fate and relative mobility in the receiving environment. Suspended particulate matter was sampled using sediment traps at several sites downstream of effluent outfall plumes as well as at reference upstream sites. Particulate metal in operationally defined fractions--exchangeable/carbonates, reducible, oxidisable and residual--were determined in suspended particulate matter with a series of selective chemical extractions. Metal enrichment in suspended particles was generally observed in both mining and urban effluent discharges. When compared to its receiving environment, the mining effluent appeared to release more particulate metals (Cu, Fe, Zn) in the most reactive fractions (i.e. exchangeable/carbonates + reducible forms, 23-43%), while other released metals, such as Cd and Mn, were predominantly in the least reactive forms (i.e., oxidisable + residual, 73-97%). In contrast, the reactivity of all particulate metals, with the exception of Mn, from the urban effluent was much higher, with up to 65, 42, 30 and 43% for Cd, Cu, Fe and Zn, respectively, in the two most reactive fractions. As expected in effluent dispersion plumes, parameters such as the organic carbon, Fe oxide and carbonate contents have specific effects on the partitioning of several trace metals, particularly Cd, Cu and Zn. Our results indicated that the relative distributions of metals among geochemical fractions varied in the effluent receiving waters where organic carbon and Fe oxides appeared as the most important parameters. This could therefore decrease the exposure for aquatic organisms that are exposed to those contaminated sediments as well as the risk to human health.

Toxicity and metal speciation in acid mine drainage treated by passive bioreactors.

Sulfate-reducing passive bioreactors treat acid mine drainage (AMD) by increasing its pH and alkalinity and by removing metals as metal sulfide precipitates. In addition to discharge limits based on physicochemical parameters, however, treated effluent is required to be nontoxic. Acute and sublethal toxicity was assessed for effluent from 3.5-L column bioreactors filled with mixtures of natural organic carbon sources and operated at different hydraulic retention times (HRTs) for the treatment of a highly contaminated AMD. Effluent was first tested for acute (Daphnia magna and Oncorhynchus mykiss) and sublethal (Pseudokirchneriella subcapitata, Ceriodaphnia dubia, and Lemna minor) toxicity. Acute toxicity was observed for D. magna, and a toxicity identification evaluation (TIE) procedure was then performed to identify potential toxicants. Finally, metal speciation in the effluent was determined using ultrafiltration and geochemical modeling for the interpretation of the toxicity results. The 10-d HRT effluent was nonacutely lethal for O. mykiss but acutely lethal for D. magna. The toxicity to D. magna, however, was removed by 2 h of aeration, and the TIE procedure suggested iron as a cause of toxicity. Sublethal toxicity of the 10-d HRT effluent was observed for all test species, but it was reduced compared to the raw AMD and to a 7.3-d HRT effluent. Data regarding metal speciation indicated instability of both effluents during aeration and were consistent with the toxicity being caused by iron. Column bioreactors in operation for more than nine months efficiently improved the physicochemical quality of highly contaminated AMD at different HRTs. The present study, however, indicated that design of passive treatment should include sufficient HRT and posttreatment aeration to meet acute toxicity requirements.

Development and validation of a chronic copper biotic ligand model for Ceriodaphnia dubia.

A biotic ligand model (BLM) to predict chronic Cu toxicity to Ceriodaphnia dubia was developed and tested. The effect of cationic competition, pH and natural organic matter complexation of Cu was examined to develop the model. There was no effect of cationic competition using increasing Ca and Na concentrations in our exposures. However, we did see a significant regression of decreasing toxicity (measured as the IC25; concentration at which there was a 25% inhibition of reproduction) as Mg concentration increased. However, taking into account the actual variability of the IC25 and since the relative increase in IC25 due to additional Mg was small (1.5-fold) Mg competition was not included in the model. Changes in pH had a significant effect on Cu IC25, which is consistent with proton competition as often suggested for acute BLMs. Finally, natural organic matter (NOM) was added to exposures resulting in significant decreases in toxicity. Therefore, our predictive model for chronic Cu toxicity to C. dubia includes the effect of pH and NOM complexation. The model was validated with Cu IC25 data generated in six natural surface waters collected from across Canada. Using WHAM VI, we calculated Cu speciation in each natural water and using our model, we generated "predicted" IC25 data. We successfully predicted all Cu IC25 within a factor of 3 for the six waters used for validation.

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).

Metal bioavailability to phytoplankton--applicability of the biotic ligand model.

To elicit a biological response from a target organism and/or to accumulate within this organism, a metal must first interact with a cell membrane. For hydrophilic metal species, this interaction with the cell surface can be represented in terms of the formation of M-X-cell surface complexes, e.g. M(z+)+(-)X-cell<-->M-X-cell, where -X-cell is a cellular ligand present at the cell surface. According to the free-ion model, or its derivative the biotic ligand model (BLM), the biological response elicited by the metal will be proportional to [M-X-cell]. In this paper, using freshwater algae as our test species, we examine some of the key assumptions that underlie the BLM, namely that metal internalization is slow relative to the other steps involved in metal uptake (i.e. the M-X-cell complex is in equilibrium with metal species in solution), that internalization occurs via cation transport, and that internalization must occur for toxicity to appear. Recent experiments with freshwater algae are described, demonstrating anomalously high metal accumulation and/or toxicity in the presence of a common low molecular weight metabolite (alanine), or in the presence of an assimilable inorganic anion (thiosulfate). The possible implications of these findings for the application of the BLM to higher organisms are discussed.