Trophic transfer of gold nanoparticles from Euglena gracilis or Chlamydomonas reinhardtii to Daphnia magna.

Understanding the trophic transfer of nanoparticles (NPs) is important because NPs are small enough to easily penetrate into organisms. In this study, we evaluated the trophic transfer of gold NPs (AuNPs) within the aquatic food chain. We observed AuNPs transfer from 2 species of primary producers (Chlamydomonas reinhardtii or Euglena gracilis) to the primary consumer (Daphnia magna). Also, bioaccumulation of AuNPs in E. gracilis was higher than that in C. reinhardtii. The reasons for the difference in Au accumulation may be the physical structure of these organisms, and the surface area that is available for interaction with NPs. C. reinhardtii has a cell wall that may act as a barrier to the penetration of NPs. The size of E. gracilis is larger than that of C. reinhardtii. This study demonstrates the trophic transfer of AuNPs from a general producer to a consumer in an aquatic environment.

Phytotoxicity of arsenic compounds on crop plant seedlings.

The effects of inorganic and organic arsenic on the germination and seedling growth of 10 crop plants were investigated to elucidate the relationship between toxicity and the arsenic chemical states. Two types of soils, soil A and B, were also tested to determine how physicochemical properties of soils were related to toxicity of arsenic and the sensitivity of the plants. All tested plant species, except mung bean and cucumber, showed inhibition of germination by two types of inorganic arsenic, arsenite, and arsenate, while the organic arsenic compound, dimethylarsinic acid (DMA), had no inhibitory effects on plants in soil A. In contrast, the growth of seedlings of all 10 plant species was sensitive to the presence of arsenic. The sensitivity of the plants toward inorganic arsenic compounds showed similar trends but differed for DMA. Overall, seedling growth was a more sensitive endpoint to arsenic toxicity than germination, and the relative toxicity of arsenic compounds on plants was arsenite > DMA > arsenate. Interestingly, the sensitivity of wheat varied significantly when the soil was changed, and the DMA was most toxic rather than arsenite in soil B. Thus, the systematic study employed here provides insights into the mechanisms of arsenic toxicity in different plant species and the role of physicochemical properties of soils.

Combined toxicities of methyl tert-butyl ether and its metabolite tert-butyl alcohol on earthworms via different exposure routes.

Methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA) are among the major soil contaminants that threaten the health of soil ecosystems. Many MTBE-contaminated sites accumulate TBA, because TBA is the intermediate of MTBE biodegradation. To access the risk of MTBE and TBA in soil, we investigated the combined toxicities of MTBE and TBA using two earthworm species, Perionyx excavatus and Eisenia andrei, as well as the toxic effects via different exposure routes. The combined toxicity showed weak antagonistic effects (LC50mix values were slightly greater than 1.0), and sensitivity toward same pollutants differed in the two earthworm species. Moreover, the toxicity of MTBE and TBA was also affected by the exposure route; both filter paper and artificial soil tests showed that dermal-only exposure to MTBE had an even greater toxic effect than combined dermal and oral exposure. Thus, we suggest that diverse environmental factors including organic materials, the physicochemical properties of the contact media, and the exposure routes of the organism, should be taken into consideration when assessing the effects of pollutants on organisms in diverse environmental systems.

Evidence of three-level trophic transfer of quantum dots in an aquatic food chain by using bioimaging.

In this study, we demonstrated the three-level trophic transfer of quantum dots (QDs) within the aquatic food chain. Using bioimaging, we observed QD transfer from protozoa (Astasia longa) to zooplankton (Moina macrocopa) to fish (Danio rerio). Bioimaging is an effective tool that can improve our understanding of the delivery of nanomaterials in vivo. Measurement with an intravital multiphoton laser scanning microscope visually proved the transfer of QDs from the first to the second and the second to the third levels. As QDs may be passed from lower organisms to humans via the food chain, our findings have implications for the safety of their use.

Conducting a battery of bioassays for gold nanoparticles to derive guideline value for the protection of aquatic ecosystems.

Gold nanoparticles (Au-NPs) are used in many applications, including the manufacture of products like cosmetics, paints, and electrochemical immunosensors, and in the detection, diagnosis, and treatment of tumors. However, there are no legal or recommended guidelines for protecting aquatic ecosystems from Au-NPs. In this study, we conducted a battery of bioassays and present toxicity values for two bacteria, one alga, one euglena, three cladoceran, and two fish species that were exposed to Au-NPs. Guideline values for protecting aquatic ecosystems from Au-NPs were derived using methods that are generally used to derive water-quality guidelines and are used in Australia, New Zealand, Canada, the European Community (EC), and the USA. Au-NPs had adverse effects on all test species, including growth inhibition of both bacteria, the alga, and the euglena; mortality and immobilization in the three cladocerans; and developmental malformations in the embryos and larvae of the two fish. Guideline values of 0.15 and 0.04 × 10(10) particles/mL were derived for Au-NPs using a species sensitivity distribution (SSD) and assessment factor. The guideline value derived for Au-NPs using an assessment factor was more stringent than that derived using SSD. This is the first study to derive guideline values for nanoparticles in water environments.

Interaction of citrate-coated silver nanoparticles with earthworm coelomic fluid and related cytotoxicity in Eisenia andrei.

Understanding the interaction of nanoparticles with biological fluid is important for predicting the behavior and toxicity of nanoparticles in living systems. The earthworm Eisenia andrei was exposed to citrate-coated silver nanoparticles (cAgNPs), and the interaction of cAgNPs with earthworm coelomic fluid (ECF), the cytotoxicity of cAgNPs in earthworm coelomocytes was assessed. The neutral red retention assay showed a reduction in lysosomal stability after exposure. The toxicity of silver ions dissolved from cAgNPs in the soil medium was not significant. The aggregation and dissolution of cAgNPs increased in ECF, which contains various electrolytes that alter the properties of nanoparticles, and their subsequent toxicity. Microscopic and dissolution studies demonstrated that the aggregation of cAgNPs rapidly increased, and readily dissolved in ECF. The bioavailability of cAgNPs to earthworms induced lysosomal cytotoxicity. This is the first report to test the interaction and lysosomal cytotoxicity of nanoparticles in earthworm biofluids.

Derivation of guideline values for gold (III) ion toxicity limits to protect aquatic ecosystems.

This study focused on estimating the toxicity values of various aquatic organisms exposed to gold (III) ion (Au(3+)), and to propose maximum guideline values for Au(3+) toxicity that protect the aquatic ecosystem. A comparative assessment of methods developed in Australia and New Zealand versus the European Community (EC) was conducted. The test species used in this study included two bacteria (Escherichia coli and Bacillus subtilis), one alga (Pseudokirchneriella subcapitata), one euglena (Euglena gracilis), three cladocerans (Daphnia magna, Moina macrocopa, and Simocephalus mixtus), and two fish (Danio rerio and Oryzias latipes). Au(3+) induced growth inhibition, mortality, immobilization, and/or developmental malformations in all test species, with responses being concentration-dependent. According to the moderate reliability method of Australia and New Zealand, 0.006 and 0.075 mg/L of guideline values for Au(3+) were obtained by dividing 0.33 and 4.46 mg/L of HC5 and HC50 species sensitivity distributions (SSD) with an FACR (Final Acute to Chronic Ratio) of 59.09. In contrast, the EC method uses an assessment factor (AF), with the 0.0006 mg/L guideline value for Au(3+) being divided with the 48-h EC50 value for 0.60 mg/L (the lowest toxicity value obtained from short term results) by an AF of 1000. The Au(3+) guideline value derived using an AF was more stringent than the SSD. We recommend that more toxicity data using various bioassays are required to develop more accurate ecological risk assessments. More chronic/long-term exposure studies on sensitive endpoints using additional fish species and invertebrates not included in the current dataset will be needed to use other derivation methods (e.g., US EPA and Canadian Type A) or the "High Reliability Method" from Australia/New Zealand. Such research would facilitate the establishment of guideline values for various pollutants that reflect the universal effects of various pollutants in aquatic ecosystems. To the best of our knowledge, this is the first study to suggest guideline values for Au(3+) levels permitted to enter freshwater environments.

Zinc oxide nanoparticles delay soybean development: a standard soil microcosm study.

Soybean is an important crop and a source of food for humans and livestock. In this study, for the first time, the long-term effects of zinc oxide (ZnO) nanoparticles on the growth, development, and reproduction of soybean [Glycine max (L.) Merrill] were evaluated in a standard soil microcosm study. The soil was treated with 0, 50, or 500 mg/kg (dry weight) of ZnO nanoparticles. The growth and development of soybean plants were tracked during a cultivation period of 8-9 weeks under greenhouse conditions. Soybean development was damaged in both treatment groups, particularly in the group that received 500 mg/kg ZnO nanoparticles. In comparison with the control group, the roots and shoots of soybeans in treatment groups were shorter and had smaller surface area and volume. Furthermore, the plants in the 500 mg/kg treatment group did not form seeds. ZnO nanoparticles negatively affected the developmental stages and reproduction of soybean plants in a soil microcosm.

Effects of zinc oxide and titanium dioxide nanoparticles on green algae under visible, UVA, and UVB irradiations: no evidence of enhanced algal toxicity under UV pre-irradiation.

Some metal oxide nanoparticles are photoreactive, thus raising concerns regarding phototoxicity. This study evaluated ecotoxic effects of zinc oxide nanoparticles and titanium dioxide nanoparticles to the green algae Pseudokirchneriella subcapitata under visible, UVA, and UVB irradiation conditions. The nanoparticles were prepared in algal test medium, and the test units were pre-irradiated by UV light in a photoreactor. Algal assays were also conducted with visible, UVA or UVB lights only without nanoparticles. Algal growth was found to be inhibited as the nanoparticle concentration increased, and ZnO NPs caused destabilization of the cell membranes. We also noted that the inhibitory effects on the growth of algae were not enhanced under UV pre-irradiation conditions. This phenomenon was attributed to the photocatalytic activities of ZnO NPs and TiO2 NPs in both the visible and UV regions. The toxicity of ZnO NPs was almost entirely the consequence of the dissolved free zinc ions. This study provides us with an improved understanding of toxicity of photoreactive nanoparticles as related to the effects of visible and UV lights.

Derivation of Ecological Protective Concentration using the Probabilistic Ecological Risk Assessment applicable for Korean Water Environment: (I) Cadmium.

Probabilistic ecological risk assessment (PERA) for deriving ecological protective concentration (EPC) was previously suggested in USA, Australia, New Zealand, Canada, and Netherland. This study suggested the EPC of cadmium (Cd) based on the PERA to be suitable to Korean aquatic ecosystem. First, we collected reliable ecotoxicity data from reliable data without restriction and reliable data with restrictions. Next, we sorted the ecotoxicity data based on the site-specific locations, exposure duration, and water hardness. To correct toxicity by the water hardness, EU's hardness corrected algorithm was used with slope factor 0.89 and a benchmark of water hardness 100. EPC was calculated according to statistical extrapolation method (SEM), statistical extrapolation methodAcute to chronic ratio (SEMACR), and assessment factor method (AFM). As a result, aquatic toxicity data of Cd were collected from 43 acute toxicity data (4 Actinopterygill, 29 Branchiopoda, 1 Polychaeta, 2 Bryozoa, 6 Chlorophyceae, 1 Chanophyceae) and 40 chronic toxicity data (2 Actinopterygill, 23 Branchiopoda, 9 Chlorophyceae, 6 Macrophytes). Because toxicity data of Cd belongs to 4 classes in taxonomical classification, acute and chronic EPC (11.07 µg/l and 0.034 µg/l, respectively) was calculated according to SEM technique. These values were included in the range of international EPCs. This study would be useful to establish the ecological standard for the protection of aquatic ecosystem in Korea.

Effect of silver nanoparticles in crop plants Phaseolus radiatus and Sorghum bicolor: media effect on phytotoxicity.

Understanding some adverse effects of nanoparticles in edible crop plants is a matter of importance because nanoparticles are often released into soil environments. We investigated the phytotoxicity of silver nanoparticles (AgNPs) on the important crop plants, Phaseolus radiatus and Sorghum bicolor. The silver nanoparticles were selected for this study because of their OECD designation as a priority nanomaterial. The toxicity and bioavailability of AgNPs in the crop plant species P. radiatus and S. bicolor were evaluated in both agar and soil media. The seedling growth of test species was adversely affected by exposure to AgNPs. We found evidence of nanoparticle uptake by plants using electron microscopic studies. In the agar tests, P. radiatus and S. bicolor showed a concentration dependent-growth inhibition effect. Measurements of the growth rate of P. radiatus were not affected in the soil studies by impediment within the concentrations tested herein. Bioavailability of nanoparticles was reduced in the soil, and the dissolved silver ion effect also differed in the soil as compared to the agar. The properties of nanoparticles have been shown to change in soil, so this phenomenon has been attributed to the reduced toxicity of AgNPs to plants in soil medium. The application of nanoparticles in soil is a matter of great importance to elucidate the terrestrial toxicity of nanoparticles.

Effect of fluorescent silica nanoparticles in embryo and larva of Oryzias latipes: sonic effect in nanoparticle dispersion.

We characterized fluorescent silica nanoparticles (FNPs), which had been applied in many biological systems, in fish embryo rearing media (ERM) solution and evaluated the potential toxicity to the early development of Oryzias latipes embryos. Distribution of FNPs in embryos and larvae of O. latipes was studied by fluorescent and confocal laser scanning microscopic studies. Embryos exposed to three different concentrations of FNPs in stirred or sonicated ERM solutions were observed up to 2d after hatching. FNPs had a negligible effect on the hatchability of O. latipes embryos; however, compared to controls, more than 30% of eggs were abnormal in 10 and 50 mg FN P L(-1) solutions. We found that the toxic effect was increased in sonicated FNP solution, which seems to be related with the dissolution of FNPs in ERM solutions that could be accelerated by sonication. Further study found that the CaCl2 included in ERM solution might enhance the dissolution of the FNPs and the silicate ion released from FNPs partially contributed to larval toxicity. This study showed that some nanoparticles may not be stable in biological fluids even if they are stable in water. Dissolution factors such as sonication and cellular components should be considered in biological application of nanoparticles.

Research trends of ecotoxicity of nanoparticles in soil environment.

We are consistently being exposed to nanomaterials in direct and/or indirect route as they are used in almost all the sectors in our life. Nations across the worlds are now trying to put global regulation policy on nanomaterials. Sometimes, they are reported to be more toxic than the corresponding ion and micromaterials. Therefore, safety research of nanoparticles has huge implications on a national economics. In this study, we evaluated and analyzed the research trend of ecotoxicity of nanoparticles in soil environment. Test species include terrestrial plants, earthworms, and soil nematode. Soil enzyme activities were also discussed. We found that the results of nanotoxicity studies were affected by many factors such as physicochemical properties, size, dispersion method and test medium of nanoparticle, which should be considered when conducting toxicity researches. In particular, more researches on the effect of physico chemical properties and fate of nanoparticles on toxicity effect should be conducted consistently.

Toxicity and bioavailability of copper nanoparticles to the terrestrial plants mung bean (Phaseolus radiatus) and wheat (Triticum aestivum): plant agar test for water-insoluble nanoparticles.

Because of their insolubility in water, nanoparticles have a limitation concerning toxicity experiments. The present study demonstrated a plant agar test for homogeneous exposure of nanoparticles to plant species. The effect of Cu nanoparticles on the growth of a plant seedling was studied, and bioaccumulation of nanoparticles was investigated. All tests were conducted in plant agar media to prevent precipitation of water-insoluble nanoparticles in test units. The plant species were Phaseolus radiatus (mung bean) and Triticum aestivum (wheat). Growth inhibition of a seedling exposed to different concentrations of Cu nanoparticles was examined. Copper nanoparticles were toxic to both plants and also were bioavailable. The 2-d median effective concentrations for P. radiatus and T. aestivum exposed to Cu nanoparticles were 335 (95% confidence level, 251-447) and 570 (450-722) mg/L, respectively. Phaseolus radiatus was more sensitive than T. aestivum to Cu nanoparticles. A cupric ion released from Cu nanoparticles had negligible effects in the concentration ranges of the present study, and the apparent toxicity clearly resulted from Cu nanoparticles. Bioaccumulation increased with increasing concentration of Cu nanoparticles, and agglomeration of particles was observed in the cells using transmission-electron microscopy-energy-dispersive spectroscopy. The present study demonstrated that the plant agar test was a good protocol for testing the phytotoxicity of nanoparticles, which are hardly water soluble.

Comparative and combined toxicities of toluene and methyl tert-butyl ether to an Asian earthworm Perionyx excavatus.

An earthworm assay was used to assess the toxicity of two main gasoline components. Ecotoxicities of toluene and methyl tert-butyl ether (MTBE) were compared for Asian earthworm Perionyx excavatus separately and in combination. Sum of toxic unit (TU) at 50% mortality for the mixture (LC50mix) was estimated from the dose (TU-based)-response relationships by the Trimmed Spearman-Karber method. Toluene was shown to be about five and two times more toxic to P. excavatus than MTBE in filter paper contact test and natural soil tests, respectively. This is primarily due to the adsorption of toluene to soil particles, leading to less bioavailability of toluene to the earthworm in soil. The combined effect of toluene and MTBE was investigated using toxic unit model, and it was found to be almost additive and synergistic responses to P. excavatus in filter paper contact test and soil tests, respectively. The combined effect of toluene and MTBE in filter paper contact test was not consistent with the results in soil toxicity tests. This phenomenon may be associated with the interaction of soil salts with pollutants. Treatment with toluene and MTBE also affected the behavior and morphology of P. excavatus. It indicates that the VOCs induce metabolic and functional damages in earthworms. Combined effects of gasoline components should be taken into account to soil risk assessment.

Decreased toxicity to terrestrial plants associated with a mixture of methyl tert-butyl ether and its metabolite tert-butyl alcohol.

The influence of the main fuel oxygenate methyl tert-butyl ether (MTBE) and its key metabolite, tert-butyl alcohol (TBA), on the growth of a plant seedling was studied separately and in combination. The test plants were mung bean (Phaseolus radiatus), cucumber (Cucumis sativus), wheat (Triticum aestivum), sorghum (Sorghum bicolor), kale (Brassica alboglabra), Chinese cabbage (Brassica campestris), and sweet corn (Zea mays). The growth of all the plants was adversely affected by TBA and MTBE. The 5-d median effective concentration (EC50) for the plants exposed to MTBE and TBA were in the range of 680 to 1,000 mg MTBE/kg soil (dry wt) and 1,200 to 3,500 mg TBA/kg soil (dry wt), respectively. The relative order of the sensitivity rankings is almost the same for MTBE and TBA. Methyl tert-butyl ether is more toxic than TBA to most of the test species. Based on the EC50 values, MTBE is approximately 1.5 to 3 times more potent than TBA. The sum of the toxic unit (TU) at 50% inhibition of the mixture (EC50mix) was calculated from the dose (TU-based)-response relationships using the trimmed Spearman-Karber method. The combined effect of MTBE + TBA on the plant growth was less than additive because the EC50mix values were greater than I TU. This phenomenon may be due to the competition of MTBE and TBA in terms of their intake by plants. The combined effects of MTBE and TBA should be taken into account to assess their risk in gasoline-contaminated sites.