Molecular toxicity identification evaluation (mTIE) approach predicts chemical exposure in Daphnia magna.

Daphnia magna is a bioindicator organism accepted by several international water quality regulatory agencies. Current approaches for assessment of water quality rely on acute and chronic toxicity that provide no insight into the cause of toxicity. Recently, molecular approaches, such as genome wide gene expression responses, are enabling an alternative mechanism based approach to toxicity assessment. While these genomic methods are providing important mechanistic insight into toxicity, statistically robust prediction systems that allow the identification of chemical contaminants from the molecular response to exposure are needed. Here we apply advanced machine learning approaches to develop predictive models of contaminant exposure using a D. magna gene expression data set for 36 chemical exposures. We demonstrate here that we can discriminate between chemicals belonging to different chemical classes including endocrine disruptors and inorganic and organic chemicals based on gene expression. We also show that predictive models based on indices of whole pathway transcriptional activity can achieve comparable results while facilitating biological interpretability.

Potential risks of effluent from acid mine drainage treatment plants at abandoned coal mines.

The lethal and sublethal toxicity of effluent from three acid mine drainage treatment plants were monitored from August 2009 to April 2010 using Daphnia magna (reference species) and Moina macrocopa (indigenous species). Acute lethal toxicity was observed in Samma effluent due to incomplete neutralization of acid mine drainages by the successive alkalinity producing system (SAPS). Additionally, there was no significant difference in toxicity values (TU) between D. magna and M. macrocopa (p < 0.05). Toxicity identification results of the final effluent collected in January 2010 showed that Al and Zn were key toxicants in addition to acidic pH. Unlike the Samma effluent, both Hwangji and Hamtae effluent had pH values that were near neutrality and showed either no acute toxicity or toxicity values less than 1 TU. However, the feeding rates of D. magna and M. macrocopa were significantly reduced when compared to the control (p < 0.05). These findings suggest that the Hamtae and Hwangji effluent likely have a sublethal effect on aquatic organisms in receiving water bodies.

A comparative study on toxicity identification of industrial effluents using Daphnia magna.

In this study, acute toxicity monitoring and toxicity identification evaluation procedures were applied to identify causative toxicants in industrial effluents. Effluents from a metal plating factory and a rubber products factory were acutely toxic toward Daphnia magna and the toxicity varied over different sampling events (2.9-5.9 and 1.7-7.6 TU, respectively). For the rubber products effluent, it was confirmed that zinc (5.65-13.18 mg L(-1)) was found to be a major cause of toxicity, which is likely originated from zinc 2-mercaptobenzothiazole and zinc diethyldithiocarbamate used as vulcanization accelerators. For the metal plating effluent, it appeared that the presence of high concentrations of Cl(-) and SO(4)(2-) (8,539-11,400 and 3,588-4,850 mg L(-1), respectively) caused the observed toxicity. These toxicants likely originated from sodium bisulfate (NaHSO(3)) and sodium hypochlorite (NaOCl) used as reducing and oxidizing agents. Though copper was found to be present in levels much higher than the EC(50) (50% effective concentration) value, this was not attributable to the toxicity of metal plating effluent likely due to complexation with dissolved organic matter.

A toxicity monitoring study on identification and reduction of toxicants from a wastewater treatment plant.

In this study, toxicity of effluents in a wastewater treatment plant and of receiving water in an adjacent stream was periodically monitored from November 2007 to June 2008, in order to trace and reduce sources of toxicants. The results showed that toxicity of final effluent (FE) changed greatly over different sampling events, and appeared to have impacts on toxicity of downstream water with a significant correlation (r(2)=0.87, p<0.05). In particular, FE toxicity was always higher than that of secondary effluent (SE). Toxicity identification evaluation (TIE) for the FE sample collected in March 2008 showed that FE toxicity was attributed to low quality of Fenton reagent with Zn contamination used for SE treatment. Furthermore, Zn concentrations in FE samples significantly correlated with FE toxicity during the sampling period (r(2)=0.95, p<0.05). After changing the Fenton reagent to one containing low Zn, Zn concentration and toxicity of FE greatly decreased in the following months.

Quantification of differentially expressed genes in Daphnia magna exposed to rubber wastewater.

In this study, differentially expressed genes (DEGs) were investigated in Daphnia magna exposed to rubber wastewater using an annealing control primer (ACP)-based polymerase chain reaction (PCR) and real-time PCR. Among three identified DEGs, two genes (DEG1 and DEG2) were up-regulated, and DEG1 expression was well-correlated to a logarithm of rubber wastewater concentration (r2=0.971, p<0.0001). In addition, DEG1 expression in D. magna exposed to rubber wastewater was strongly correlated with that of D. magna exposed to Zn (r2=0.9513, p<0.05), suggesting that the induction of DEG1 was caused by Zn, which is the dominant toxicant in rubber wastewater. In addition, DEG1 expression was more sensitive to toxicants than immobility, which is the conventional endpoint in toxicity tests using D. magna. The lowest observed effect concentrations (LOEC) determined using immobility tests were 2.5% for rubber wastewater and 1.6mgl(-1) for Zn. In contrast, a significant increase in DEG1 expression was observed at exposure concentrations of as low as 0.6% rubber wastewater and 0.2mgl(-1) Zn. These results indicate that DEG1 is a sensitive and quantitative biomarker of water and wastewater containing Zn.

Toxicity identification and reduction of wastewaters from a pigment manufacturing factory.

In this study, major toxicants in pigment wastewaters (i.e. raw wastewater and effluent) were determined by toxicity identification evaluations (TIEs), and changes in their toxicities due to combined gamma-ray (gamma-ray) and ozone treatments were evaluated. From TIE results using Daphnia magna, Cr(VI) and o-toluidine were identified as major toxicants in the raw wastewater with concentrations of 2.47 and 25.3mg l(-1), respectively. This was confirmed by subsequent spiking and mass balance approaches. For effluent, TIE results suggested that organic anions were likely responsible for the observed toxicities. Toxicity reduction tests showed that gamma-rays with ozone treatment after coagulation was more efficient in reducing toxicities in the raw wastewater (3.9-2.0 TU) than gamma-ray treatment alone (3.9-3.1 TU). For pigment effluent, gamma-ray/O3 treatment resulted in a complete removal of the acute toxicity.

Toxicity identification in metal plating effluent: implications in establishing effluent discharge limits using bioassays in Korea.

Because of complexity and diversity of toxicants in effluent, chemical analysis alone gives very limited information on identifying toxic chemicals to test organisms. Toxicity identification evaluation (TIE) techniques have been widely used to identify toxicants in various samples including industrial wastewater as well as natural waters. In response to new regulation for effluent discharge in Korea, which will be effective from 2011, a necessity of studies emerges that investigates toxicity levels in industrial effluents. This work was a preliminary study examining toxicity levels in effluent from one metal plating factory using Daphnia magna (48 h immobility) and identifying toxicity-causing substances. Toxicity tests showed variability on different sampling occasions and the results of TIE methods indicated that both organic compounds and metals contributed to the observed toxicity in metal plating effluent. Further studies are necessary to help reduce effluent toxicity especially from direct dischargers, who will have to comply with the new regulation.

Acute toxicity of Ag and CuO nanoparticle suspensions against Daphnia magna: the importance of their dissolved fraction varying with preparation methods.

A variety of methods to prepare nanoparticle suspensions have been employed for aquatic toxicity tests, although they can influence the dispersion property and subsequent toxicity of nanoparticles. Thus, in this study, we prepared stock suspensions of silver (Ag) and copper oxide (CuO) nanoparticles using different methods and compared their acute toxicity against Daphnia magna. The results showed that the dispersion method, filtration and initial concentration largely affected their toxicity, when the toxicity was expressed as the total concentrations of Ag and Cu. In case of Ag nanoparticles, the toxicity was also influenced by their different particle size. However, negligible differences in 24h-median effect concentration (EC(50)) values, which were calculated in terms of their dissolved concentrations, were observed. When expressing toxicity on the basis of dissolved concentrations, 24h-EC(50) values of the Ag and CuO nanoparticles were also found to be similar to those of the counterpart ionic species, i.e., Ag (as AgNO(3)) and Cu (as CuCl(2)·2H(2)O). These findings indicate that the dissolved fraction of nanoparticles largely contributes to their acute toxicity. Our results may help in establishing a useful guideline for preparing nanoparticle suspensions with reproducible toxicity.

Identification of multi-level toxicity of liquid crystal display wastewater toward Daphnia magna and Moina macrocopa.

Toxicity-based regulations of industrial effluent have been adopted to complement the conventional discharge limits based on chemical analyses. In this study, multi-level toxicity including acute toxicity, feeding rate inhibition and oxidative stress of effluent from a liquid crystal display (LCD) wastewater treatment plant (WWTP) to Daphnia magna (reference species) and Moina macrocopa (native species) were periodically monitored from April 2010 to April 2011. Raw wastewater was acutely toxic to both D. magna and M. macrocopa, but the toxicity reached less than 1 TU in the final effluent (FE) as treatment proceeded. Although acute toxicity was not observed in the FE, the feeding rate of daphnids was significantly inhibited. Additionally, the antioxidant enzyme activity of catalase, superoxide dismutase and glutathione peroxidase (GPx) in D. magna increased significantly when compared to the control, while only GPx activity was increased significantly in M. macrocopa (p<0.05). A toxicity identification evaluation using D. magna showed that Cu was the key toxicant in the FE, which was not effectively removed by the coagulation/flocculation process in the LCD WWTP. In addition, Al originating from the coagulant seemed to increase toxicity of the FE.

Using response surface methodology to assess the effects of iron and spent mushroom substrate on arsenic phytotoxicity in lettuce (Lactuca sativa L.).

The effects of iron (Fe) and spent mushroom substrate (SMS) arsenic (As) phytotoxicity towards lettuce in artificial soils were investigated to separate the adverse soil parameters relating to As toxicity using a response surface methodology. SMS induced the root elongation of lettuce in both control and As-treated soils. However, in phytotoxicity test using a median effective concentration (EC(50)) of As, Fe and the interaction between both parameters (Fe*SMS) significantly affected EC(50), which explained 71% and 23% of the response, respectively. The refined model was as follows: EC(50) of As (mgkg(-1))=10.99+60.03 × Fe-10.50 × Fe*SMS. The results confirmed that the soil parameters relating to the As mobility in soils were important factors affecting its toxicity. In conclusion, Fe significantly reduced the As phytotoxicity. However, although SMS enhanced the root elongation, SMS in As-treated soils decreased EC(50) of As on the root growth via its interaction with Fe. Despite the limitations of the artificial soils and range of parameters studied, the application of this statistical tool can be considered a powerful and efficient technique for interpretation and prediction of the complicated results caused by the interactions between many factors within the soil environments.

Combined effects of water quality parameters on mixture toxicity of copper and chromium toward Daphnia magna.

In this study, a central composite design (CCD) was employed to evaluate the combined effects of pH, hardness and dissolved organic carbon (DOC) on the toxicity of a mixture of Cu(II) and Cr(VI) toward Daphnia magna. Overall, the results showed that increases in pH, hardness and DOC concentration led to decreased mixture toxicity of Cu(II) and Cr(VI) by reducing the concentrations of toxic species such as Cu(2+) and HCrO(4)(-). In addition, empirical models for the prediction of 24-h and 48-h mortalities of D. magna were developed and validated by using three different sources of dissolved organic matter (DOM). Because the DOMs had different Cu(II)-binding capacities, the empirical models were revised using the ligand concentration of DOMs instead of the DOC concentration; however, the prediction capability of these models did not differ significantly. These results suggest that it is not likely that the chemical property of DOM is important for prediction of the mixture toxicity of Cu(II) and Cr(VI) toward D. magna when the ligand concentration of DOMs greatly exceed the Cu(II) concentration.

Quantification of water solubilized CdSe/ZnS quantum dots in Daphnia magna.

The relative transparency of Daphnia magna (daphnia) and the unique optical properties of quantum dots (QDs) were paired to study the accumulation potential and surface coating effects on uptake of amphiphilic polymer coated CdSe/ZnS QDs. Fluorescence confocal laser scanning microscopy was used to visualize and spectrally distinguish QDs from competing autofluorescent signals arising from the daphnia themselves and their food sources. QDs were found to accumulate within the digestive tracts of daphnia, as well as, in some cases, adhere to the carapace, antennae, and thoracic appendages. After 48 h of gut clearance with and without feeding, QD fluorescence signal was still apparent in the digestive tracts of daphnia, and inductively coupled plasma mass spectrometry (ICP-MS) measurements confirmed that 36-53% of the initial uptake was retained. As surface charge and pegylation can influence the uptake of nanoparticles, uptake of QDs coated with two different amphililic polymers and their polyethylene glycol (PEG) coated counterparts was also examined. Fluorescence microscopy and ICP-MS measurements revealed differences in uptake after 24 h of exposure which were attributed to particle surface coating and stability.