Diatoms Reduce Decomposition of and Fungal Abundance on Less Recalcitrant Leaf Litter via Negative Priming.

Heterotrophic microbial decomposers colonize submerged leaf litter in close spatial proximity to periphytic algae that exude labile organic carbon during photosynthesis. These exudates are conjectured to affect microbial decomposers' abundance, resulting in a stimulated (positive priming) or reduced (negative priming) leaf litter decomposition. Yet, the occurrence, direction, and intensity of priming associated with leaf material of differing recalcitrance remains poorly tested. To assess priming, we submerged leaf litter of differing recalcitrance (Alnus glutinosa [alder; less recalcitrant] and Fagus sylvatica [beech; more recalcitrant]) in microcosms and quantified bacterial, fungal, and diatom abundance as well as leaf litter decomposition over 30 days in absence and presence of light. Diatoms did not affect beech decomposition but reduced alder decomposition by 20% and alder-associated fungal abundance by 40% in the treatments including all microbial groups and light, thus showing negative priming. These results suggest that alder-associated heterotrophs acquired energy from diatom exudates rather than from leaf litter. Moreover, it is suggested that these heterotrophs have channeled energy to alternative (reproductive) pathways that may modify energy and nutrient availability for the remaining food web and result in carbon pools protected from decomposition in light-exposed stream sections.

Effects of copper in Daphnia are modulated by nanosized titanium dioxide and natural organic matter: what is the impact of aging duration?

During its aquatic life cycle, nanosized titanium dioxide (nTiO2) may interact with natural organic matter (NOM) ultimately altering the ecotoxicity of co-occurring chemical stressors such as heavy metals (e.g. copper (Cu)). In this context, the following hypotheses were tested: (1) aging of nTiO2 along with Cu reduces Cu toxicity, (2) nTiO2 agglomerates have a lower potential to reduce Cu toxicity and (3) aging of nTiO2 in presence of NOM reduces Cu toxicity further. A multifactorial test design crossing three nTiO2 levels (0.0, 0.6 and 3.0 mg/L) with two levels of NOM (0 versus 8 mg total organic carbon (TOC)/L) and seven nominal Cu concentrations (ranging from 0 to 1536 µg/L) aged in ASTM medium for 0, 1, 3 and 6 days was realised, while two aging scenarios were applied (type 1: nTiO2 jointly aged with Cu; type 2: Cu added after nTiO2 aging). Subsequently, Cu toxicity was assessed using the immobility of Daphnia magna after 48 h of exposure as response variable. The experiments revealed that neither aging duration nor the extent of agglomeration (type 1 vs. type 2 aging) has a substantial impact on Cu induced toxicity. Moreover, it was confirmed that the presence of NOM substantially reduced Cu induced toxicity, independent of the aging scenario and duration. More importantly, the data suggest the ingestion of Cu loaded nTiO2 as additional exposure pathway contributing to Cu toxicity. In conclusion, it seems unlikely that nTiO2 concentrations currently detected in or predicted for aquatic ecosystems, which are at least one order of magnitude below the concentration tested here, influence Cu toxicity meaningfully.

The importance of diet-related effects of the antibiotic ciprofloxacin on the leaf-shredding invertebrate Gammarus fossarum (Crustacea; Amphipoda).

Antibiotics may constitute a risk for aquatic detritivorous macroinvertebrates (i.e., shredders) via waterborne and dietary antibiotic exposure. In addition, antibiotics can alter the food quality for shredders mediated by shifts in leaf-associated decomposer (i.e., aquatic fungi and bacteria) communities. However, little is known about the relative importance of the waterborne and dietary effect pathway. Therefore, we followed a tiered testing approach aimed at assessing the relative importance of these effect pathways. We employed the antibiotic ciprofloxacin (CIP) and the shredder Gammarus fossarum as model stressor and test species, respectively. In a first step, we assessed the short-term waterborne toxicity of CIP using survival and leaf consumption of G. fossarum as response variables. Alterations in the leaf-associated decomposer community, which may be reflected by their palatability, were assessed using food choice assays. Finally, we conducted a 2 × 2-factorial experiment over 24 days assessing the pathways individually and combined using energy processing (i.e., leaf consumption and feces production), growth and energy storage (i.e., neutral lipid fatty acids) as variables. Short term waterborne exposure indicated low toxicity with LC50 and EC50 values of 13.6 and 6.4 mg CIP/L, respectively. At the same time, shredders did not prefer any leaf material during the food choice assay. However, the fungal community was significantly affected in the highest CIP-treatments (0.5 and 2.5 mg/L) suggesting an altered food quality for shredders. This assumption is supported by the results of the long-term assay. At 0.5 mg CIP/L, gammarids' leaf consumption, growth and energy storage were increased when subjected via the dietary pathway, which was linked to changes in the leaf-associated microbial community. Our data highlight the importance of dietary effect pathways for effects on shredders, potentially impacting energy dynamics in detritus-based stream ecosystems.

Reduction of Pesticide Toxicity Under Field-Relevant Conditions? The Interaction of Titanium Dioxide Nanoparticles, Ultraviolet, and Natural Organic Matter.

In surface waters, the illumination of photoactive engineered nanomaterials (ENMs) with ultraviolet (UV) light triggers the formation of reactive intermediates, consequently altering the ecotoxicological potential of co-occurring organic micropollutants including pesticides due to catalytic degradation. Simultaneously, omnipresent natural organic matter (NOM) adsorbs onto ENM surfaces, altering the ENM surface properties. Also, NOM absorbs light, reducing the photo(cata)lytic transformation of pesticides. Interactions between these environmental factors impact 1) directly the ecotoxicity of photoactive ENMs, and 2) indirectly the degradation of pesticides. We assessed the impact of field-relevant UV radiation (up to 2.6 W UVA/m²), NOM (4 mg TOC/L), and photoactive ENM (nTiO2, 50 µg/L) on the acute toxicity of 6 pesticides in Daphnia magna. We selected azoxystrobin, dimethoate, malathion, parathion, permethrin, and pirimicarb because of their varying photo- and hydrolytic stabilities. Increasing UVA alone partially reduced pesticide toxicity, seemingly due to enhanced degradation. Even at 50 µg/L, nano-sized titanium dioxide (nTiO2) reduced but also increased pesticide toxicity (depending on the applied pesticide), which is attributable to 1) more efficient degradation and potentially 2) photocatalytically induced formation of toxic by-products. Natural organic matter 1) partially reduced pesticide toxicity, not evidently accompanied by enhanced pesticide degradation, but also 2) inhibited pesticide degradation, effectively increasing the pesticide toxicity. Predicting the ecotoxicological potential of pesticides based on their interaction with UV light or interaction with NOM was hardly possible, which was even more difficult in the presence of nTiO2. Environ Toxicol Chem 2020;39:2237-2246. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

The fate of silver nanoparticles in riverbank filtration systems - The role of biological components and flow velocity.

Riverbank filtration is a natural process that may ensure the cleaning of surface water for producing drinking water. For silver nanoparticles (AgNP), physico-chemical interaction with sediment surfaces is one major retention mechanism. However, the effect of flow velocity and the importance of biological retention, such as AgNP attachment to biomass, are not well understood, yet. We investigated AgNP (c = 0.6 mg L-1) transport at different spatial and temporal scales in pristine and previously pond water-aged sediment columns. Transport of AgNP under near-natural conditions was studied in a long-term riverbank filtration experiment over the course of one month with changing flow scenarios (i.e. transport at 0.7 m d-1, stagnation, and remobilization at 1.7 m d-1). To elucidate retention processes, we conducted small-scale lab column experiments at low (0.2 m d-1) and high (0.7 m d-1) flow rate using pristine and aged sediments. Overall, AgNP accumulated in the upper centimeters of the sediment both in lab and outdoor experiments. In the lab study, retention of AgNP by attachment to biological components was very effective under high and low flow rate with nearly complete NP accumulation in the upper 2 mm. When organic material was absent, abiotic filtration mechanisms led to NP retention in the upper 5 to 7 cm of the column. In the long-term study, AgNP were transported up to a depth of 25 cm. For the pristine sediment in the lab study and the outdoor experiments only erratic particle breakthrough was detected in a depth of 15 cm. We conclude that physico-chemical interactions of AgNP with sediment surfaces are efficient in retaining AgNP. The presence of organic material provides additional retention sites which increase the filtration capacity of the system. Nevertheless, erratic breakthrough events might transport NP into deeper sediment layers.

Nanoparticles transported from aquatic to terrestrial ecosystems via emerging aquatic insects compromise subsidy quality.

Nanoparticle contaminants enter aquatic ecosystems and are transported along the stream network. Here, we demonstrate a novel pathway for the return of nanoparticles from aquatic to terrestrial ecosystems via cross-boundary subsidies. During their emergence, trichopteran caddisflies carried titanium dioxide and gold nanoparticles into their terrestrial life stages. Moreover, their emergence was delayed by ≤30 days, and their energy reserves were depleted by ≤25%. Based on worst case estimates, it is suggested that terrestrial predators, such as bats feeding on aquatic prey, may ingest up to three orders of magnitude higher gold levels than anticipated for humans. Additionally, terrestrial predator species may suffer from alterations in the temporal availability and nutritional quality of their prey. Considering the substantial transfer of insect biomass to terrestrial ecosystems, nanoparticles may decouple aquatic and terrestrial food webs with important (meta-)ecosystem level consequences.

Exposure pathway dependent effects of titanium dioxide and silver nanoparticles on the benthic amphipod Gammarus fossarum.

The increasing production of engineered inorganic nanoparticles (EINPs) elevates their release into aquatic ecosystems raising concerns about associated environmental risks. Numerous investigations indicate sediments as the final sink, facilitating the exposure of benthic species to EINPs. Although reports of sub-lethal EINP effects on benthic species are increasing, the importance of exposure pathways (either waterborne or dietary) is poorly understood. This study investigates the influence of two EINPs, namely titanium dioxide (nTiO2) and silver (nAg), on the benthic model organism Gammarus fossarum specifically addressing the relative relevance of these pathways. For each type of EINP an individual 30-day long bioassay was conducted, applying a two-factorial test design. The factors include the presence or absence of the EINPs (nTiO2: ∼80 nm, 4 mg/L or nAg: ∼30 nm, 0.125 mg/L; n = 30) in the water phase (waterborne), combined with a preceding 6-day long aging of their diet (black alder leaves) also in presence or absence of the EINPs (dietary). Response variables were mortality, food consumption, feces production and energy assimilation. Additionally, the physiological fitness was examined using lipid content and dry weight of the organisms as measures. Results revealed a significantly reduced energy assimilation (up to ∼30%) in G. fossarum induced by waterborne exposure towards nTiO2. In contrast, the dietary exposure towards nAg significantly increased the organisms' energy assimilation (up to ∼50%). Hence, exposure pathway dependent effects of EINPs cannot be generalized and remain particle specific resting upon their intrinsic properties affecting their potential to interact with the surrounding environment. As a result of the different properties of the EINPs used in this study, we clearly demonstrated variations in type and direction of observed effects in G. fossarum. The results of the present study are thus supporting current approaches for nano-specific grouping that might enable an enhanced accuracy in predicting EINP effects facilitating their environmental risk assessment.

A blessing in disguise? Natural organic matter reduces the UV light-induced toxicity of nanoparticulate titanium dioxide.

Besides their economic value, engineered inorganic nanoparticles (EINPs) may pose a risk for the integrity of ecosystems. Among EINPs, titanium dioxide (nTiO2) is frequently used and released into surface waters in the µg range. There, nTiO2 interacts with environmental factors, influencing its potential to cause adverse effects on aquatic life. Although factors like ultra violet (UV) light and natural organic matter (NOM) are considered as ubiquitous, their joint impact on nTiO2-induced toxicity is poorly understood. This study addressed the acute toxicity of nTiO2 (P25; 0.00-64.00 mg/L; ~60 nm) at ambient UV light (0.00-5.20 W UVA/m2) and NOM levels (seaweed extract; 0.00-4.00 mg TOC/L), using the immobility of Daphnia magna as response variable. Confirming previous studies, effects caused by nTiO2 were elevated with increasing UV radiation (up to ~280 fold) and mitigated by higher NOM levels (up to ~12 fold), possibly due to reduced reactive oxygen species (ROS; measured as •OH radicals) formation at lower UV intensities. However, contradicting to former studies, nTiO2-mediated ROS formation was not proportional to increasing NOM levels: lower concentrations (0.04-0.40 mg TOC/L) slightly diminished, whereas a higher concentration (4.00 mg TOC/L) promoted the ROS quantity, irrespective of UV intensity. Measured ROS levels do not fully explain the observed nTiO2-induced toxicity, whereas increasing acetylcholinesterase and glutathione-S-transferase activities in daphnids (in presence of 8.00 mg/L nTiO2 and elevated UV intensity) point towards neurotoxic and oxidative stress as a driver for the observed effects. Hence, despite higher •OH levels in the treatments where 4.00 mg TOC/L were present, NOM was still capable of reducing nTiO2-induced stress and ultimately adverse effects in aquatic life.

SETAC GLB and SETAC Europe SAC: a liaison promoting the next generation of ecotoxicologists and environmental chemists.

This commentary is an introduction for students to the Society of Environmental Toxicology and Chemistry (SETAC) and its Student Advisory Council (SAC). As young academics face challenges while trying to develop their careers, SETAC and the SAC help facilitate student involvement in the various communities within the society that can help to develop the students' careers within the environmental sciences [e.g. the German Language Branch (GLB)]. This piece would also like to emphasize and pay homage to the continual cooperation between the SAC and the ESEU, which provides a scientific platform to communicate internationally and beyond the borders of SETAC, as well as offer heartfelt congratulations from the SAC to the GLB for their "20 Years SETAC GLB" and deep gratitude for their strong advocacy and support of the SAC.

Assessing the effects of field-relevant pesticide mixtures for their compliance with the concentration addition model - An experimental approach with Daphnia magna.

The environmental risk assessment of pesticides is mainly performed on individual active ingredients. In surface waters within the agricultural landscape, however, contamination is usually characterized by complex pesticide mixtures. To estimate the joint effects caused by these complex mixtures, mathematical models have been proposed. Among these, the model of concentration addition (CA) is suggested as default model for the risk assessment of chemical mixtures as it is considered protective for mixtures composed of similar and dissimilar acting substances. Here we assessed the suitability of CA predictions for seven field relevant pesticide mixtures using acute (immobility) and chronic (reproduction) responses of the standard test species Daphnia magna. Pesticide mixtures indicated largely additive or less than additive effects when using CA model predictions as a reference. Moreover, we revealed that deviations from CA predictions are lower for chronic (up to 3.2-fold) relative to acute (up to 7.2-fold) response variables. Additionally, CA predictions were in general more accurate for complex mixtures relative to those composed of only a few pesticides. Thus, this study suggests CA models as largely protective for the risk assessment of pesticide mixtures justifying its use as default model. At the same time, extrapolating conclusions about the joint effects of pesticides from acute to chronic responses is uncertain, due to partly large discrepancies with regards to the deviation of model prediction and observed effects between exposure scenarios.

The evil within? Systemic fungicide application in trees enhances litter quality for an aquatic decomposer-detritivore system.

Waterborne exposure towards fungicides is known to trigger negative effects in aquatic leaf-associated microbial decomposers and leaf-shredding macroinvertebrates. We expected similar effects when these organisms use leaf material from terrestrial plants that were treated with systemic fungicides as a food source since the fungicides may remain within the leaves when entering aquatic systems. To test this hypothesis, we treated black alder (Alnus glutinosa) trees with a tap water control or a systemic fungicide mixture (azoxystrobin, cyprodinil, quinoxyfen, and tebuconazole) at two worst-case application rates. Leaves of these trees were used in an experiment targeting alterations in two functions provided by leaf-associated microorganisms, namely the decomposition and conditioning of leaf material. The latter was addressed via the food-choice response of the amphipod shredder Gammarus fossarum. During a second experiment, the potential impact of long-term consumption of leaves from trees treated with systemic fungicides on G. fossarum was assessed. Systemic fungicide treatment altered the resource quality of the leaf material resulting in trends of increased fungal spore production and an altered community composition of leaf-associated fungi. These changes in turn caused a significant preference of Gammarus for microbially conditioned leaves that had received the highest fungicide treatment over control leaves. This higher food quality ultimately resulted in a higher gammarid growth (up to 300% increase) during the long-term feeding assay. Although the underlying mechanisms still need to be addressed, the present study demonstrates a positive indirect response in aquatic organisms due to systemic pesticide application in a terrestrial system. As the effects from the introduction of plant material treated with systemic fungicides strongly differ from those mediated via other pathways (e.g., waterborne exposure), our study provides a novel perspective of fungicide-triggered effects in aquatic detritus-based food webs.

Nanoparticles in the environment: where do we come from, where do we go to?

Nanoparticles serve various industrial and domestic purposes which is reflected in their steadily increasing production volume. This economic success comes along with their presence in the environment and the risk of potentially adverse effects in natural systems. Over the last decade, substantial progress regarding the understanding of sources, fate, and effects of nanoparticles has been made. Predictions of environmental concentrations based on modelling approaches could recently be confirmed by measured concentrations in the field. Nonetheless, analytical techniques are, as covered elsewhere, still under development to more efficiently and reliably characterize and quantify nanoparticles, as well as to detect them in complex environmental matrixes. Simultaneously, the effects of nanoparticles on aquatic and terrestrial systems have received increasing attention. While the debate on the relevance of nanoparticle-released metal ions for their toxicity is still ongoing, it is a re-occurring phenomenon that inert nanoparticles are able to interact with biota through physical pathways such as biological surface coating. This among others interferes with the growth and behaviour of exposed organisms. Moreover, co-occurring contaminants interact with nanoparticles. There is multiple evidence suggesting nanoparticles as a sink for organic and inorganic co-contaminants. On the other hand, in the presence of nanoparticles, repeatedly an elevated effect on the test species induced by the co-contaminants has been reported. In this paper, we highlight recent achievements in the field of nano-ecotoxicology in both aquatic and terrestrial systems but also refer to substantial gaps that require further attention in the future.

History Matters: Pre-Exposure to Wastewater Enhances Pesticide Toxicity in Invertebrates.

Disturbance regimes determine communities' structure and functioning. Nonetheless, little effort has been undertaken to understand interactions of press and pulse disturbances. In this context, leaf-shredding macroinvertebrates can be chronically exposed to wastewater treatment plant effluents (i.e., press disturbance) before experiencing pesticide exposure following agricultural runoff (i.e., pulse disturbance). It is assumed that wastewater pre-exposure alters animals' sensitivity to pesticides. To test this hypothesis, we exposed model-populations of the shredder Gammarus fossarum to wastewater at three field-relevant dilution levels (i.e., 0%, 50%, and 100%). After 2, 4, and 6 weeks, survival, leaf consumption, dry weight, and energy reserves were monitored. Additionally, animals were assessed for their sensitivity toward the neonicotinoid insecticide thiacloprid using their feeding rate as response variable. Both wastewater treatments reduced gammarids' survival, leaf consumption, dry weight, and energy reserves. Moreover, both wastewater pre-exposure scenarios increased animals' sensitivity toward thiacloprid by up to 2.5 times compared to the control. Our results thus demonstrate that press disturbance as posed by wastewater pre-exposure can enhance susceptibility of key players in ecosystem functioning to further (pulse) disturbances. Therefore, applying mitigation measures such as advanced treatment technologies seems sensible to support functional integrity in the multiple-stress situation.

Long-term effects of fungicides on leaf-associated microorganisms and shredder populations-an artificial stream study.

Leaf litter is a major source of carbon and energy for stream food webs, while both leaf-decomposing microorganisms and macroinvertebrate leaf shredders can be affected by fungicides. Despite the potential for season-long fungicide exposure for these organisms, however, such chronic exposures have not yet been considered. Using an artificial stream facility, effects of a chronic (lasting up to 8 wk) exposure to a mixture of 5 fungicides (sum concentration 20 µg/L) on leaf-associated microorganisms and the key leaf shredder Gammarus fossarum were therefore assessed. While bacterial density and microorganism-mediated leaf decomposition remained unaltered, fungicide exposure reduced fungal biomass (≤71%) on leaves from day 28 onward. Gammarids responded to the combined stress from consumption of fungicide-affected leaves and waterborne exposure with a reduced abundance (≤18%), which triggered reductions in final population biomass (18%) and in the number of precopula pairs (≤22%) but could not fully explain the decreased leaf consumption (19%), lipid content (≤43%; going along with an altered composition of fatty acids), and juvenile production (35%). In contrast, fine particulate organic matter production and stream respiration were unaffected. Our results imply that long-term exposure of leaf-associated fungi and shredders toward fungicides may result in detrimental implications in stream food webs and impairments of detrital material fluxes. These findings render it important to understand decomposer communities' long-term adaptational capabilities to ensure that functional integrity is safeguarded. Environ Toxicol Chem 2017;36:2178-2189. © 2017 SETAC.

Quantity and quality of natural organic matter influence the ecotoxicity of titanium dioxide nanoparticles.

Nanoparticles' fate is amongst other parameters determined by the quantity and quality of natural organic matter (NOM). Consequently, the ecotoxicity of nanoparticles is modified, while only little information is available on the NOM characteristics triggering this interplay. This study systematically examined how NOM quantity and quality influences the acute ecotoxicity of titanium dioxide nanoparticles (nTiO2) towards Daphnia magna. Therefore, two nTiO2 products (A-100 and P25; ∼100 nm) were investigated in combination with seven NOM types of variable quality at four levels each (up to 4.00 mg total organic carbon/L). The results showed that - independent of the applied nTiO2 product and NOM type - nTiO2 ecotoxicity decreased up to a factor of >18 with increasing NOM concentration. More importantly, increasing levels of aromaticity and hydrophobicity of the NOM decreased the magnitude of toxic effects caused by nTiO2, which was again independent of the nTiO2 product tested. In the light of the ubiquitary presence of NOM, the ecotoxicological risk of nTiO2 in surface waters with high NOM loads is likely moderate. However, interactions of nTiO2 and NOM in combination with other natural or chemical stressors are not well-understood but seem to be fundamental for a reliable risk assessment of nanoparticles.

Palladium Nanoparticles: Is There a Risk for Aquatic Ecosystems?

Nano-sized palladium (nano-Pd) is used in catalytic converters of automobiles, where it can be released into the environment by abrasion. Although these particles may subsequently be transported into surface water bodies, no data estimating their fate and toxicity in aquatic systems exists. This study characterized the particle size development of nano-Pd (advertised size ~12 nm; hydrodynamic size ~70 nm) in media with variable ionic strength (IS). Additionally, the particles' acute toxicity for daphnids and chironomids was assessed. While nano-Pd agglomerated more quickly with increasing IS, it caused only marginal effects in both test species after 96 h of exposure. After 144 h of exposure, however, an EC50 value of 1.23 mg nano-Pd/L for daphnids was determined indicating effects over the long run. When considering the relatively low environmental concentration of elemental Pd in surface waters (usually ng/L), though, this study suggests only a low aquatic risk in response to nano-Pd.

Does the presence of titanium dioxide nanoparticles reduce copper toxicity? A factorial approach with the benthic amphipod Gammarus fossarum.

In aquatic ecosystems, titanium dioxide nanoparticles (nano-TiO2) may adsorb co-occurring chemical stressors, such as copper (Cu). This interaction has the potential to reduce the concentration of dissolved Cu due to surface binding to the nanoparticles. The subsequent sedimentation of nano-TiO2 agglomerates may increase the exposure of benthic species towards the associated Cu. This scenario was assessed by employing the amphipod Gammarus fossarum as model species and taking advantage of a 2×2-factorial design investigating absence and presence of 2mg nano-TiO2/L and 40µg Cu/L (n=45; t=24d) in darkness, respectively. Nano-TiO2 alone did not affect mortality and leaf consumption, whereas Cu alone caused high mortality (>70%), reduced leaf consumption (25%) and feces production (30%) relative to the control. In presence of nano-TiO2, Cu-induced toxicity was largely eliminated. However, independent of Cu, nano-TiO2 decreased the gammarids' assimilation and weight. Hence, nano-TiO2 may be applicable as Cu-remediation agent, while its potential long-term effects need further attention.

Aging of TiO2 Nanoparticles Transiently Increases Their Toxicity to the Pelagic Microcrustacean Daphnia magna.

During their aquatic life cycle, nanoparticles are subject to environmentally driven surface modifications (e.g. agglomeration or coating) associated with aging. Although the ecotoxicological potential of nanoparticles might be affected by these processes, only limited information about the potential impact of aging is available. In this context, the present study investigated acute (96 h) and chronic (21 d) implications of systematically aged titanium dioxide nanoparticles (nTiO2; ~90 nm) on the standard test species Daphnia magna by following the respective test guidelines. The nTiO2 were aged for 0, 1, 3 and 6 d in media with varying ionic strengths (Milli-Q water: approx. 0.00 mmol/L and ASTM: 9.25 mmol/L) in the presence or absence of natural organic matter (NOM). Irrespective of the other parameters, aging in Milli-Q did not change the acute toxicity relative to an unaged control. In contrast, 6 d aged nTiO2 in ASTM without NOM caused a fourfold decreased acute toxicity. Relative to the 0 d aged particles, nTiO2 aged for 1 and 3 d in ASTM with NOM, which is the most environmentally-relevant setup used here, significantly increased acute toxicity (by approximately 30%), while a toxicity reduction (60%) was observed for 6 d aged nTiO2. Comparable patterns were observed during the chronic experiments. A likely explanation for this phenomenon is that the aging of nTiO2 increases the particle size at the start of the experiment or the time of the water exchange from <100 nm to approximately 500 nm, which is the optimal size range to be taken up by filter feeding D. magna. If subjected to further agglomeration, larger nTiO2 particles, however, cannot be retained by the daphnids' filter apparatus ultimately reducing their ecotoxicological potential. This non-linear pattern of increasing and decreasing nTiO2 related toxicity over the aging duration, highlights the knowledge gap regarding the underlying mechanisms and processes. This understanding seems, however, fundamental to predict the risks of nanoparticles in the field.