Understanding PFAS toxicity through cell culture metabolomics: Current applications and future perspectives.

Per- and polyfluoroalkyl substances (PFAS), ubiquitous environmental contaminants, pose significant challenges to ecosystems and human health. While cell cultures have emerged as new approach methodologies (NAMs) in ecotoxicity research, metabolomics is an emerging technique used to characterize the small-molecule metabolites present in cells and to understand their role in various biological processes. Integration of metabolomics with cell cultures, known as cell culture metabolomics, provides a novel and robust tool to unravel the complex molecular responses induced by PFAS exposure. In vitro testing also reduces reliance on animal testing, aligning with ethical and regulatory imperatives. The current review summarizes key findings from recent studies utilizing cell culture metabolomics to investigate PFAS toxicity, highlighting alterations in metabolic pathways, biomarker identification, and the potential linkages between metabolic perturbations. Additionally, the paper discusses different types of cell cultures and metabolomics methods used for studies of environmental contaminants and particularly PFAS. Future perspectives on the combination of metabolomics with other advanced technologies, such as single-cell metabolomics (SCM), imaging mass spectrometry (IMS), extracellular flux analysis (EFA), and multi-omics are also explored, which offers a holistic understanding of environmental contaminants. The synthesis of current knowledge and identification of research gaps provide a foundation for future investigations that aim to elucidate the complexities of PFAS-induced cellular responses and contribute to the development of effective strategies for mitigating their adverse effects on human health.

Sediment spiking and equilibration procedures to achieve partitioning of uranium similar to contamination in tropical wetlands near a mine site.

The derivation of sediment quality guideline values (SQGVs) presents significant challenges. Arguably the most important challenge is to conduct toxicity tests using contaminated sediments with physico-chemistry that represents real-world scenarios. We used a novel metal spiking method for an experiment that ultimately aims to derive a uranium SQGV. Two pilot studies were conducted to inform the final spiking design, i.e. percolating a uranyl sulfate solution through natural wetland sediments. An initial pilot study that used extended mixing equilibration phases produced hardened sediments not representative of natural sediments. A subsequent percolation method produced sediment with similar texture to natural sediment and was used as the method for spiking the sediments. The range of total recoverable uranium (TR-U) concentrations achieved was 8-3200 mg/kg. This reflected the concentrations found in natural wetlands and water management ponds found on a uranium mine site and was above natural levels. Dilute-acid extractable uranium (AE-U) concentrations were >80% of total concentrations, indicating that much of the uranium in the spiked sediment was labile and potentially bioavailable. The portion of TR-U extractable as AE-U was similar at the start and end of the 4.5-month field-deployment. Porewater uranium (PW-U) analyses indicated that partition coefficients (Kd) were 2000-20,000 L/kg, and PW-U was greater in post- than pre-field-deployed samples when TR-U was ≤1500 mg/kg, indicating the binding became weaker during the field-deployment period. At higher spiked-U concentrations, the PW-U was lower post-field-deployment. Comparing the physico-chemical data of the spiked sediments with environmental monitoring data from sediments in the vicinity of a uranium mining operation indicated that they were representative of sediments contaminated by mining and that the U-spiked sediments had a clear U concentration gradient. This confirmed the suitability of the spiking procedure for preparing sediments that were suitable for deriving a SQGV for uranium.

Translocation of pharmaceuticals from wastewater into beehives.

There has been a substantial research focus on the presence of pesticides in flowers and the subsequent exposure to honeybees. Here we demonstrate for the first time that honeybees can also be exposed to pharmaceuticals, commonly present in wastewater. Residues of carbamazepine (an anti-epileptic drug) up to 371 ng/mL and 30 µg/g were detected in nectar and pollen sampled from zucchini flowers (Cucurbita pepo) grown in carbamazepine spiked soil (0.5-20 µg/g). Under realistic exposure conditions from the use of recycled wastewater, carbamazepine concentrations were estimated to be 0.37 ng/L and 30 ng/kg in nectar and pollen, respectively. Incorporation of environmentally relevant carbamazepine residues in nectar and pollen into a modelling framework able to simulate beehive dynamics including the honeybee foraging activity at the landscape scale (BEEHAVE and BEESCOUT) enabled the simulation of carbamazepine translocation from zucchini fields into honeybee hives. Carbamazepine accumulation was modelled in 11 beehives across a 25 km2 landscape over three years chosen to represent distinct climatic conditions. During a single flowering period, carbamazepine concentrations were simulated to range between 0 and 2478 ng per beehive. The amount of carbamazepine gathered not only varied across the simulated years but there were also differences in accumulation of carbamazepine between beehives within the same year. This work illustrates a fundamental first step in assessing the risk of pharmaceuticals to bees through realistic scenarios by demonstrating a method to quantify potential exposure of honeybees at the landscape scale. Pharmaceuticals are being inadvertently but increasingly applied to agricultural lands globally via the use of wastewater for agricultural irrigation in response to water scarcity problems. We have demonstrated a route of pharmaceutical exposure to honeybees via contaminated nectar and pollen. Given the biological potency of pharmaceuticals, accumulation of these chemicals in nectar and pollen suggest potential implications for honeybee health, with unknown ecosystem consequences.