Potential of the Red Alga Dixoniella grisea for the Production of Additives for Lubricants.

There is an increasing interest in algae-based raw materials for medical, cosmetic or nutraceutical applications. Additionally, the high diversity of physicochemical properties of the different algal metabolites proposes these substances from microalgae as possible additives in the chemical industry. Among the wide range of natural products from red microalgae, research has mainly focused on extracellular polymers for additive use, while this study also considers the cellular components. The aim of the present study is to analytically characterize the extra- and intracellular molecular composition from the red microalga Dixoniella grisea and to evaluate its potential for being used in the tribological industry. D. grisea samples, fractionated into extracellular polymers (EPS), cells and medium, were examined for their molecular composition. This alga produces a highly viscous polymer, mainly composed of polysaccharides and proteins, being secreted into the culture medium. The EPS and biomass significantly differed in their molecular composition, indicating that they might be used for different bio-additive products. We also show that polysaccharides and proteins were the major chemical compounds in EPS, whereas the content of lipids depended on the separation protocol and the resulting product. Still, they did not represent a major group and were thus classified as a potential valuable side-product. Lyophilized algal fractions obtained from D. grisea were found to be not toxic when EPS were not included. Upon implementation of EPS as a commercial product, further assessment on the environmental toxicity to enchytraeids and other soil organisms is required. Our results provide a possible direction for developing a process to gain an environmentally friendly bio-additive for application in the tribological industry based on a biorefinery approach.

Ion compositions in artificial media control the impact of humic acid on colloidal behaviour, dissolution and speciation of CuO-NP.

The toxicity of copper oxide nanoparticles (CuO-NP) strongly depends on their interactions with the surrounding environment, impacting their dissolution and colloidal stability. This behaviour is studied quite extensively for simplified electrolytes, but information on the behaviour of CuO-NP in more complex artificial media are lacking. In our study, we analysed the colloidal behaviour and considered the speciation of CuO-NP in pure water and three artificial media of different complexity which are used in ecotoxicology. Measurements were done over 7 days in the absence and presence of humic acid (HA) as a model organic molecule. In pure water, the addition of HA lowered the zeta potential from +11 to -41 mV, while in all artificial media, it stayed constantly at about -20 mV. The hydrodynamic diameter of CuO-NP remained unaffected by HA in pure water and seawater, while in porewater and especially in freshwater, HA suppressed strong agglomeration. In pure water, HA strongly increased dissolution to the highest observed value (3% of total Cu), while HA reduced dissolution in all artificial media. Speciation calculations revealed that cations from the media competed with Cu from the NP surface for complexing sites of the HA. This competition may have caused the reduced dissolution in the presence of ions. Furthermore, speciation calculations also suggest that ion composition drove agglomeration behaviour rather than ion concentration: agglomeration was high when divalent cations where the major interaction partner and dominant in relative terms. HA may have reduced the relative dominance and thus altered the agglomeration, aligning it in all media. Summarizing, ion composition and the presence of HA strongly drive the dissolution and agglomeration of CuO-NP in artificial media, consequently, analysing complexation can help to predict environmental behaviour and toxicity.

How test vessel properties affect the fate of silver nitrate and sterically stabilized silver nanoparticles in two different test designs used for acute tests with Daphnia magna.

The relation between test conditions such as medium composition or pH on silver nanoparticle (AgNP) behavior and its link to toxicity is one of the major topics in nanoecotoxicological research in the last years. In addition, the adaptation of the ecotoxicological standard tests for nanomaterials is intensely discussed to increase comparability and reliability of results. Due to the limitation of test material production volumes and the need for high-throughput screening, miniaturization has been proposed for several test designs. In the present study, the effect of a miniaturization of the acute Daphnia immobilization test on AgNP behavior was investigated. For this purpose, available, adsorbed, and dissolved silver fractions were measured using AgNP and silver nitrate in the following two test designs: a standard test (ST) design and a miniaturized test (MT) design with reduced test volume and less animals. Despite the increase in surface area in relation to the test volume in MT, more AgNP attached to the ST vessel surface, so that in this case, exposure concentrations were significantly lower compared to the MT assessment. Ionic silver concentrations resulting from AgNP dissolution were similar in both test designs. The same was observed for ionic silver concentrations in silver nitrate (AgNO3) treatments, but adsorbed silver was also higher in ST treatments. Assessing the structure-activity relationships revealed that surface properties such as hydrophobicity, potential binding sites, or surface roughness were of higher importance than surface:volume ratios for both test substances.

Behavior and chronic toxicity of two differently stabilized silver nanoparticles to Daphnia magna.

While differences in silver nanoparticle (AgNP) colloidal stability, surface potential, or acute aquatic toxicity for differently stabilized AgNP have often been reported, these have rarely been studied in long-term ecotoxicity tests. In the current study, we investigated the chronic toxicity of AgNP to Daphnia magna over a 21-day period with two different stabilizers (citrate and detergent), representative for charge and sterical stabilizers, respectively. This was coupled with a series of short-term experiments, such as mass balance and uptake/depuration testing, to investigate the behavior of both types of AgNP during a typical media exchange period in the D. magna test for chronic toxicity. As expected, the sterically stabilized AgNP was more stable in the test medium, also in the presence of food; however, a higher uptake of silver after 24h exposure of the charge stabilized AgNP was found compared to the detergent-stabilized AgNP (0.046±0.006µgAgµgDW(-1) and 0.023±0.005µgAgµgDW(-1), respectively). In accordance with this, the higher reproductive effects and mortality were found for the charge-stabilized than for the sterically-stabilized silver nanoparticles in 21-d tests for chronic toxicity. LOEC was 19.2µgAgL(-1) for both endpoints for citrate-coated AgNP and >27.5µgAgL(-1) (highest tested concentration for detergent-stabilized AgNP). This indicates a link between uptake and toxicity. The inclusion of additional short-term experiments on uptake and depuration is recommended when longer-term chronic experiments with nanoparticles are conducted.

Adaptation of the Daphnia sp. acute toxicity test: miniaturization and prolongation for the testing of nanomaterials.

Manufacturing of nanomaterials (NMs) is often complex and expensive, and their environmental risks are poorly understood or even unknown. An economization of testing NMs is therefore desirable, which can be achieved by miniaturizing test systems. However, the downsizing of test vessels and volumes can enlarge the surface/volume ratio (SVR) which in turn can affect the bioavailable concentration of adsorbing substances like NMs. The present study focused on the miniaturization of the acute toxicity test with Daphnia magna. The adaptations were verified with three reference substances, the non-adsorbing potassium dichromate (K2Cr2O7) and as potentially highly-adsorbing substances silver nanoparticles (AgNPs) and silver nitrate (AgNO3). The miniaturized test was conducted in 24-well microtiter plates (MT) and simultaneously compared to the OECD standard test (ST). Furthermore, the test duration was prolonged from 48 to 96 h since NMs tend to show effects only after extended exposure. The toxicity of K2Cr2O7 and AgNPs continued to increase within the prolonged test span. The test comparisons with K2Cr2O7 did not reveal any significant differences between ST and MT. AgNO3 toxicity was significantly decreased in MT compared to ST due to the enlarged SVR. The toxicity of AgNPs in MT after 24 h was equal to ST. Contrary to our expectations an exposure longer than 24 h resulted in an increase of AgNP toxicity in MT, possibly due to enhanced dissolution of silver. Microtiter plates are appropriate alternative test vessels for the Daphnia sp. acute toxicity test; thus, its miniaturization is feasible. The enlarged SVR has to be taken into account since it can affect the toxicity of potentially adsorbing substances. Furthermore, the standard test duration of 48 h might underestimate the toxicity of many substances, especially of NMs.