Katja Knauer

Nanomaterials in Plant Protection and Fertilization: Current State, Foreseen Applications, and Research Priorities

Scientific publications and patents on nanomaterials (NM) used in plant protection or fertilizer products have exponentially increased since the millennium shift. While the United States and Germany have published the highest number of patents, Asian countries released most scientific articles. About 40% of all contributions deal with carbon-based NM, followed by titanium dioxide, silver, silica, and alumina. Nanomaterials come in many diverse forms (surprisingly often ≫100 nm), from solid doped particles to (often nonpersistent) polymer and oil-water based structures. Nanomaterials serve equally as additives (mostly for controlled release) and active constituents. Product efficiencies possibly increased by NM should be balanced against enhanced environmental NM input fluxes. The dynamic development in research and its considerable public perception are in contrast with the currently still very small number of NM-containing products on the market. Nanorisk assessment and legislation are largely in their infancies.

Are Carbon Nanotube Effects on Green Algae Caused by Shading and Agglomeration

Due to growing production, carbon nanotubes (CNT) may soon be found in a broad range of products and thus in the environment. In this work, an algal growth test was developed to determine effects of pristine and oxidized CNT on the green algae Chlorella vulgaris and Pseudokirchneriella subcapitata. CNT suspensions were prepared in algal test medium and characterized taking into account the suspension age, the reduced light transmittance of nanoparticle suspensions defined as shading of CNT and quantified by UV/vis spectroscopy, and the agglomeration of the CNT and of the algal cells. Growth inhibition and photosynthetic activity were investigated as end points. Growth of C. vulgaris was inhibited with effect concentrations of 50% (EC(50)) values of 1.8 mg CNT/L and of 24 mg CNT/L in well dispersed and in agglomerated suspensions, respectively, and 20 mg CNT/L and 36 mg CNT/L for P. subcapitata, respectively. However, the photosynthetic activity was not affected. Growth inhibition was highly correlated with the shading of CNT and the agglomeration of algal cells. This suggests that the reduced algal growth might be caused mainly by indirect effects, i.e. by reduced availability of light and different growth conditions caused by the locally elevated algal concentration inside of CNT agglomerates.

ADSORPTION AND UPTAKE OF COPPER BY THE GREEN ALGA SCENEDESMUS SUBSPICATUS (CHLOROPHYTA)1

Copper (II) accumulation has been investigated in the green alga Scenedesmus subspicatus G. Brinkmann considering both adsorption and uptake kinetics. Experiments were conducted in a Cu‐ and PH‐buffered medium at different free Cu2+ concentrations that were neither growth limiting nor toxic. We distinguished between adsorption on the cell surface and intracellular uptake by extracting copper from the cells with EDTA. Data from short‐term experiments were compared with data obtained from experiments under steady state conditions. The accumulation of Cu can be described by two processes, an initial fast adsorption occurring within a minute followed by a slower intracellular uptake. Metal uptake followed Michaelis‐Menten kinetics and is mediated by two systems, one with low and the other with high affinity. The maximum uptake rates (1.30 × 10−‐10 mol·[g dry wt algae]−1· min−1, 3.67 × 10−‐9 mol·[g dry wt algae]−1·min−1), and the half‐saturation constants (6.84 × 10−‐14 M, 2.82 × 10−‐12 M) for the two uptake systems were determined using the Lineweaver‐Burk plot. The calculated maximum concentration of binding sites on the surface of the algae is initially higher (9.0 × 10−‐6 mol Cu.[g dry wt algae]−1) than under steady state conditions (2.9 × 10−‐6 mol Cu·[g dry wt algae]−1). This suggests that the initial binding to the algal surface comprises the binding to specific transport ligands as well as to inert adsorption sites. The conditional stability constant of the Cu binding to surface ligands was calculated as log KCu= 11.0 at pH 7.9. This freshwater alga has a high ability to accumulate Cu, reflecting its adaptation to the bioavailable concentration of copper.

Diuron sorbed to carbon nanotubes exhibits enhanced toxicity to Chlorella vulgaris.

Carbon nanotubes (CNT) are more and more likely to be present in the environment, where they will associate with organic micropollutants due to strong sorption. The toxic effects of these CNT-micropollutant mixtures on aquatic organisms are poorly characterized. Here, we systematically quantified the effects of the herbicide diuron on the photosynthetic activity of the green alga Chlorella vulgaris in presence of different multiwalled CNT (industrial, purified, pristine, and oxidized) or soot. The presence of carbonaceous nanoparticles reduced the adverse effect of diuron maximally by <78% (industrial CNT) and <34% (soot) at 10.0 mg CNT/L, 5.0 mg soot/L, and diuron concentrations in the range 0.73-2990 μg/L. However, taking into account the measured dissolved instead of the nominal diuron concentration, the toxic effect of diuron was equal to or stronger in the presence of CNT by a factor of up to 5. Sorbed diuron consequently remained partially bioavailable. The most pronounced increase in toxicity occurred after a 24 h exposure of algae and CNT. All results point to locally elevated exposure concentration (LEEC) in the proximity of algal cells associated with CNT as the cause for the increase in diuron toxicity.

Effects of photosystem II inhibitors and their mixture on freshwater phytoplankton succession in outdoor mesocosms

Effects of three photosystem II inhibitors and of their mixture on a freshwater phytoplankton community were studied in outdoor mesocosms. Atrazine, isoproturon, and diuron were applied as 30% hazardous concentrations (HC30s) obtained from species-sensitivity distributions. Taking concentration addition into account, the mixture comprised one-third of the HC30 of each substance. Effects were investigated during a five-week period of constant concentrations and a five-month posttreatment period when the herbicides dissipated. Total abundance, species composition, and diversity and recovery of the community were evaluated. Ordination techniques, such as principal component analysis and principal response curve, were applied to compare the various treatments on the community level. The three herbicides stimulated comparable effects on total abundance and diversity of phytoplankton during the period of constant exposure because of the susceptibility of the dominant cryptophytes Chroomonas acuta and Cryptomonas erosa et ovata and the prasinophyte Nephroselmis cf. olivacea. Moreover, concentration addition described combined effects of atrazine, isoproturon, and diuron on total abundance and diversity in the constant-exposure period, because their mixture induced effects on abundance and diversity similar to those of the single substances. Principal component and principal response curve analyses revealed that the community structure of diuron- and isoproturon-treated phytoplankton recovered two weeks after constant exposure, which might be related to the fast dissipation of the phenylureas. Species compositions of mixture- and atrazine-treated communities were not comparable to that of the control community five months after the end of constant exposure. This might be explained by the slower dissipation of atrazine relative to the phenylureas and by differences in the species sensitivities, resulting in a different succession of phytoplankton.

Manganese uptake and Mn(II) oxidation by the alga Scenedesmus subspicatus

Abstract: To examine the relationships among manganese concentration in the culture medium, algae growth, manganese uptake and Mn(II) oxidation on the algal surface, we exposed the unicellular alga Scenedesmus subspicatus to a broad range of free Mn2+ ion concentrations. Extra- and intracellular manganese concentrations were distinguished by reducing the Mn oxides with ascorbate. A large fraction of the Mn bound by the algae occurred as reducible Mn(III/IV) oxides formed by Mn(II) oxidation, presumably on the algal surface, although these algal cells of small size are not likely to build up a large pH gradient at their surface. Limitation of growth occurred when [Mn2+] was lower than about 1×10-9 M, which corresponded to an intracellular content of 10-7 mol Mn (g algae dry weight)-1. Inhibition of growth occurred at high [Mn2+], at which high concentrations of Mn (>1×10-5 Mol Mn (g algae dry weight)-1) were present both intracellularly and as extracellular Mn oxides. Phytoplankton collected in various lakes contained intracellular Mn and extracellular Mn oxides in similar concentrations as the Scenedesmus cultures.n

Sorption kinetics and equilibrium of the herbicide diuron to carbon nanotubes or soot in absence and presence of algae

Carbon nanotubes (CNT) are strong sorbents for organic micropollutants, but changing environmental conditions may alter the distribution and bioavailability of the sorbed substances. Therefore, we investigated the effect of green algae (Chlorella vulgaris) on sorption of a model pollutant (diuron, synonyms: 3-(3,4-Dichlorophenyl)-1,1-dimethylurea, DCMU) to CNT (multi-walled purified, industrial grade, pristine, and oxidized; reference material: Diesel soot). In absence of algae, diuron sorption to CNT was fast, strong, and nonlinear (Freundlich coefficients: 10(5.79)-10(6.24)xa0μg/kgCNT·(μg/L)(-n) and 0.62-0.70 for KF and n, respectively). Adding algae to equilibrated diuron-CNT mixtures led to 15-20% (median) diuron re-dissolution. The relatively high amorphous carbon content slowed down ad-/desorption to/from the high energy sorption sites for both industrial grade CNT and soot. The results suggest that diuron binds readily, but - particularly in presence of algae - partially reversibly to CNT, which is of relevance for environmental exposure and risk assessment.

Effects of Titanium Dioxide Nanoparticles on Red Clover and Its Rhizobial Symbiont

Titanium dioxide nanoparticles (TiO2 NPs) are in consideration to be used in plant protection products. Before these products can be placed on the market, ecotoxicological tests have to be performed. In this study, the nitrogen fixing bacterium Rhizobium trifolii and red clover were exposed to two TiO2 NPs, i.e., P25, E171 and a non-nanomaterial TiO2. Growth of both organisms individually and their symbiotic root nodulation were investigated in liquid and hydroponic systems. While 23 and 18 mg l-1 of E171 and non-nanomaterial TiO2 decreased the growth rate of R. trifolii by 43 and 23% respectively, P25 did not cause effects. Shoot length of red clover decreased between 41 and 62% for all tested TiO2 NPs. In 21% of the TiO2 NP treated plants, no nodules were found. At high concentrations certain TiO2 NPs impaired R. trifolii as well as red clover growth and their symbiosis in the hydroponic systems.

Low-dose addition of silver nanoparticles stresses marine plankton communities

The release of silver nanoparticles (AgNPs) is expected to rise in the near future, with possible negative effects on aquatic life and enhancement of microbial resistance against AgNPs. However, a realistic evaluation of the toxicity of AgNPs to the marine ecosystem is currently missing. Therefore, we designed a mesocosm experiment to assess the impact of AgNP exposure on natural microbial plankton community dynamics in a coastal marine site at environmentally relevant concentrations. We monitored changes in the composition of the planktonic community, from viruses to protists. Further, we analyzed the concentration and properties of AgNPs for the total time of exposure. We found that the addition of AgNPs even at a low dose affected the plankton communities. Specifically, the growth of Synechococcus was inhibited and bacterial community composition significantly changed. Additionally, the amount of a lysogeny-related gene increased and viral auxiliary metabolic genes that are involved in cyanobacterial photosynthesis decreased, revealing a damaged photosynthetic potential after AgNP exposure. Microbial plankton was significantly affected due to both increased dissolved silver ions and decreased AgNP size. Our results highlight that the release of AgNPs alters the functioning of the marine food web by hampering important viral and bacterial processes.