Reduced bioavailability of Au and isotopically enriched 109Ag nanoparticles transformed through a pilot wastewater treatment plant in Hyalella azteca under environmentally relevant exposure scenarios.

Wastewater Treatment Plants (WWTP) are a major repository and entrance path of nanoparticles (NP) in the environment and hence play a major role in the final NP fate and toxicity. Studies on silver nanoparticles (AgNP) transport via the WWTP system and uptake by aquatic organisms have so far been carried out using unrealistically high AgNP concentrations, unlikely to be encountered in the aquatic environment. The use of high AgNP concentrations is necessitated by both the low sensitivity of the detection methods used and the need to distinguish background Ag from spiked AgNP. In this study, isotopically enriched 109AgNP were synthesized to overcome these shortcomings and characterized by a broad range of methods including transmission electron microscopy, dynamic and electrophoretic light scattering. 109AgNP and gold NP (AuNP) were spiked to a pilot wastewater treatment plant fed with municipal wastewater for up to 21 days. AuNP were used as chemically less reactive tracer. The uptake of the pristine and transformed NP present in the effluent was assessed using the benthic amphipod Hyalella azteca in fresh- and brackish water exposures at environmentally relevant concentrations of 30 to 500 ng Au/L and 39 to 260 ng Ag/L. The unique isotopic signature of the 109AgNP allowed to detect the material at environmentally relevant concentrations in the presence of a much higher natural Ag background. The results show that the transformations reduce the NP uptake at environmentally relevant exposure concentrations. For 109Ag, lower accumulation factors (AF) were obtained after exposure to transformed NP (250-350) compared to the AF values obtained for pristine 109AgNP (750-840). The reduced AF values observed for H. azteca exposed to effluent from the AuNP-spiked WWTP indicate that biological transformation processes (e.g. eco-corona formation) seem to be involved in addition to chemical transformation.

Invertebrate Species for the Bioavailability and Accumulation Assessment of Manufactured Polymer-Based Nano- and Microplastics.

Bioaccumulation tests with invertebrates have recently been discussed as a suitable alternative to bioaccumulation tests with metal- or metal oxide-based nanoparticles in fish for regulatory assessment. In the present study, as a first step, we investigated the suitability of three invertebrate species for bioaccumulation tests with nano- and microplastics. In a laboratory approach the freshwater bivalve Corbicula fluminea, the freshwater amphipod Hyalella azteca, and the terrestrial isopod Porcellio scaber were exposed to fluorescently labeled nano- and microplastics to evaluate their suitability to estimate the bioavailability and bioaccumulation of these test items. No bioaccumulation was observed in H. azteca or P. scaber. In contrast, the measurement of the relative fluorescence of the test items in the soft tissue and the feces of the filter-feeding bivalve allowed us to derive data that may be useful for the regulatory bioaccumulation assessment of manufactured nano- and microplastics. The developed measurement method using fluorescence represents a time-efficient and cost-effective analytical method for manufactured nano- and microplastics in laboratory studies for regulatory assessment. Environ Toxicol Chem 2022;41:961-974. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Are Fragrance Encapsulates Taken Up by Aquatic and Terrestrial Invertebrate Species?

The uptake potential of fragrance encapsulates by aquatic or terrestrial organisms was investigated. Because of their size of <5 mm and their polymeric nature, fragrance encapsulates fall under the Joint Group of Experts on the Scientific Aspects of Marine Environmental Protection definition of microplastics. After use, fragrance encapsulates enter the sewer system and reach the sewage treatment plant (STP), where >90% of them are likely to be removed by sorption to the sludge. When the STP-generated sludge is used as fertilizer for agricultural soils, this may lead to potential exposure of terrestrial invertebrates to fragrance encapsulates, especially those feeding on particles of a similar size as the fragrance encapsulates. Two aquatic (Corbicula fluminea [water exposure] and Hyalella azteca [water and dietary exposure]) and one terrestrial invertebrate (Eisenia andrei [soil exposure]) species were exposed to 50 mg/L (or mg/kg) double fluorescence-labeled fragrance encapsulates (diameter 5-50 µm). The results showed that fragrance encapsulates are available to aquatic and terrestrial invertebrates but that species-specific differences regarding the ability to ingest fragrance encapsulates may exist. The benthic grazer H. azteca showed no ingestion of fragrance encapsulates, whereas the capsules were readily ingested and egested by the unselective freshwater filter feeder C. fluminea as well as the terrestrial decomposer E. andrei. No signs of bioaccumulation of fragrance encapsulates were indicated by microscopic assessment. Environ Toxicol Chem 2022;41:931-943. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Testing the bioaccumulation potential of manufactured nanomaterials in the freshwater amphipod Hyalella azteca.

Standardized experimental approaches for the quantification of the bioaccumulation potential of nanomaterials in general and in (benthic) invertebrates in particular are currently lacking. We examined the suitability of the benthic freshwater amphipod Hyalella azteca for the examination of the bioaccumulation potential of nanomaterials. A flow-through test system that allows the generation of bioconcentration and biomagnification factors was applied. The feasibility of the system was confirmed in a 2-lab comparison study. By carrying out bioconcentration and biomagnification studies with gold, titanium dioxide and silver nanoparticles as well as dissolved silver (AgNO3) we were able to assess the bioaccumulation potential of different types of nanomaterials and their exposure pathways. For this, the animals were examined for their total metal body burden using inductively coupled mass spectroscopy (ICP-MS) and for the presence of nanoparticulate burdens using single-particle ICP-MS. The role of released ions was highlighted as being very important for the bioavailability and bioaccumulation of metals from nanoparticles for both examined uptake paths examined (bioconcentration and biomagnification). In 2018 a tiered testing strategy for engineered nanomaterials was proposed by Handy et al. that may allow a waiver of bioaccumulation fish studies using inter alia invertebrates. Data gained in studies carried out with invertebrates like the developed Hyalella azteca test may be included in this proposed tiered testing strategy.

Chronic effects of wastewater-borne silver and titanium dioxide nanoparticles on the rainbow trout (Oncorhynchus mykiss).

Even though nanoparticles (NPs) are mostly removed by wastewater treatment plants, wastewater-borne NPs may show an altered toxicity to aquatic organisms. The main objectives of this work were: i) to assess the chronic (28 days) effects of wastewater-borne NPs of silver (AgNPs, 1.4-36.2 µg L-1) and titanium dioxide (TiO2NPs, 3.1-50.2 µg L-1) at the individual (growth) and biochemical (biomarkers of neurotoxicity, oxidative stress and energy metabolism) levels in rainbow trout Oncorhynchus mykiss; and ii) to compare them with their effluent-supplemented and water-dispersed counterparts. The total Ag and Ti levels were determined in several fish organs. The growth of O. mykiss was not affected by the NPs in any treatment, except a 29% increase at 5.5 µg L-1 of total Ag supplemented to effluents. The Ag level in organs of O. mykiss was significantly higher after exposure to water-dispersed AgNPs than their wastewater-borne or effluent-supplemented counterparts. No significant Ti uptake could be observed. Effluent-supplemented TiO2NPs (50.1 µg L-1 Ti) potentially induced neurotoxic effects, indicated by a 24% increase in acetylcholinesterase activity comparatively to controls. Energy reserves were unaffected by TiO2 treatments, while nearly all AgNP-containing treatments caused a depletion of total lipids, proteins and carbohydrates in the muscle, suggesting an increased energy demand for detoxification processes to cope with AgNPs. Besides NPs, the effluent matrix and dispersing agent (for AgNPs) induced significant effects on energetic reserves and oxidative stress, indicating background toxicity of both treatments at the biochemical level. Our study is the first to assess chronic effects of wastewater-borne NPs on rainbow trout. While no effects were found at the individual level, several biochemical markers were changed by the NPs exposure. Our results highlight the importance of using complex matrices for a reliable risk assessment of NPs in the aquatic environment.

Bioconcentration, Metabolism, and Spatial Distribution of 14 C-Labeled Laurate in the Freshwater Amphipod Hyalella azteca.

Regulatory assessment of the bioaccumulation from water is commonly based on bioconcentration factors (BCFs) derived from fish flow-through tests. Such experiments require many laboratory animals and are time-consuming and costly. An alternative test setup for organic, neutral compounds using the amphipod Hyalella azteca was recently suggested, resulting in BCF values which show a strong correlation with fish BCF data. In the present study, the bioconcentration potential of the ionic compound laurate was elucidated in H. azteca. The sodium salt of 1-14 C laurate was applied to H. azteca in a flow-through and a semistatic approach. Because of rapid biodegradation, a semistatic approach with frequent medium replacements was required to ensure a stable medium concentration. Laurate was also rapidly metabolized by H. azteca. A large proportion of the total radioactivity measured in the amphipod tissue was not extractable, suggesting that mineralized laurate was accumulated in the calcified exoskeleton of H. azteca. This was confirmed in a further study using carbonate [14 C]. A lipid-normalized (5.0%) Hyalella BCF of 8.9 was calculated for laurate, measured as free fatty acids. The results of the bioconcentration studies with H. azteca confirm the low bioaccumulation potential of the test item previously observed in fish. However, more organic ionic compounds with various properties need to be tested to assess whether a general correlation between fish and Hyalella BCF data exists. Environ Toxicol Chem 2020;39:310-322. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.