Passive Dosing of Petroleum and Essential Oil UVCBs-Whole Mixture Toxicity Testing at Controlled Exposure.

Petroleum products and essential oils are produced and used in large amounts and are categorized as "Substances of Unknown or Variable composition, Complex reaction products or Biological materials (UVCBs)." These UVCBs are notorious difficult-to-test substances, since they are complex mixtures of hydrophobic and volatile compounds. This study introduces two passive dosing (PD) approaches for whole UVCB toxicity testing: (1) headspace PD applies the UVCB and purified lipid oil as a donor to control exposure via the headspace and (2) silicone rod PD applies UVCB-loaded silicone rods to control exposure via an aqueous test medium and headspace. Headspace gas chromatography-mass spectrometry measurements were used to cross-validate the approaches at the saturation level and to confirm exposure and maintain mixture composition at varying donor concentration levels. Both approaches were applied to whole-mixture toxicity tests of petroleum and essential oil UVCBs with daphnia and algae. Finally, the observed toxicity was linked to concentrations in the donor and in lipid membranes at equilibrium with the donors. Dose-response curves were similar across the dosing approaches and tested species for petroleum products but differed by an order of magnitude between essential oils and PD systems. All observed toxic effects were consistent with baseline toxicity, and no excess mixture toxicity was observed.

Surface-Related Toxicity of Polystyrene Beads to Nematodes and the Role of Food Availability.

Microplastics released into freshwaters from anthropogenic sources settle in the sediments, where they may pose an environmental threat to benthic organisms. However, few studies have considered the ecotoxicological hazard of microplastic particles for nematodes, one of the most abundant taxa of the benthic meiofauna. This study investigated the toxic effects of polystyrene (PS) beads (0.1-10.0 µm) and the underlying mechanisms thereof on the reproduction of the nematode Caenorhabditis elegans. The observed effect of the PS beads on the nematodes correlated well with the total surface area of the beads per volume, with a 50% inhibition of reproduction at 55.4 ± 12.9 cm2/mL, independent of the bead size. The adverse effects were not explained by styrene monomers leaching from the beads because chemical activities of styrene in PS suspensions were well below the toxic levels. However, the observed effects could be related to the bead material because the same-sized silica (SiO2) beads had considerably less impact, probably due to their higher specific density. PS and SiO2 beads affected the food availability of C. elegans, with greater effects by the PS beads. Our results demonstrate the importance of including indirect food web effects in studies of the ecological risks posed by microplastics.

Effects of α-pinene on life history traits and stress tolerance in the springtail Folsomia candida.

Volatile monoterpenes are emitted in large quantities to both air and soil by many plant species. While studies have addressed effects of monoterpenes on aboveground invertebrates, we have much poorer understanding of the possible effects of monoterpenes on soil invertebrates. Monoterpenes play a protective role in some plant species during heat and water stress, and therefore may provide similar protection against abiotic stress to soil invertebrates. The aim of the present study was to investigate the effects of the common monoterpene, α-pinene, on the soil living springtail, Folsomia candida (Collembola; Isotomidae). We hypothesized that exposure to α-pinene would lower the transition temperature of membranes, and thereby improve cold tolerance. Controlled exposure to α-pinene, which is a volatile liquid at room temperature, was made possible by passive dosing through the air-phase using a lipid donor. This lipid-based passive dosing approach also allows linking observed effects to concentrations in membrane when equilibrium is achieved. Equilibrium membrane concentrations above 116 mmol kg-1 caused springtails to become comatose, and coma recovery time was proportional to exposure concentration. Alpha-pinene delayed time to first egg laying, while the number of eggs laid and hatchability was unaffected. Springtails exposed to α-pinene showed increased survival of cold shock (-6 °C, 2 h), but no effects on heat (34 °C, 2 h) or drought tolerance (98.2% relative humidity, 7d) were observed. The present study has demonstrated that α-pinene has direct toxic effects to F. candida, but on the other hand can improve their cold tolerance considerably at membrane concentrations above 87 mmol kg-1.

Headspace Passive Dosing of Volatile Hydrophobic Organic Chemicals from a Lipid Donor-Linking Their Toxicity to Well-Defined Exposure for an Improved Risk Assessment.

High hydrophobicity and volatility of chemicals often lead to substantial experimental challenges but were here utilized in headspace passive dosing (HS-PD) to establish and maintain exposure: the pure chemical served as a passive dosing donor for controlling exposure at saturation, whereas triglyceride oil containing the chemical was used to control lower exposure levels. These donor solutions were added to glass inserts placed in the closed test systems. Mass balance calculations confirmed a dominant donor capacity for all chemicals except isooctane. This HS-PD method was applied to algal growth inhibition and springtail lethality tests with terpenes, alkanes, and cyclic siloxanes. Headspace concentrations above the lipid donors were measured for three chemicals to determine their chemical activity, using saturated vapor as the analytical standard and thermodynamic reference. Toxicity was related to chemical activity and calculated concentrations in membranes at equilibrium with the lipid donor. For both tests and all chemicals, toxic effects were observed within or above the reported range for baseline toxicity, meaning that no excess toxicity was observed. The toxicity of siloxanes was markedly higher to the terrestrial springtail than the aquatic algae, which is consistent with a more efficient mass transfer of these volatile hydrophobic chemicals in air compared to water.

Headspace passive dosing of volatile hydrophobic chemicals - Aquatic toxicity testing exactly at the saturation level.

It is challenging to conduct aquatic tests with highly hydrophobic and volatile chemicals while avoiding substantial sorptive and evaporative losses. A simple and versatile headspace passive dosing (HS-PD) method was thus developed for such chemicals: The pure liquid test chemical was added to a glass insert, which was then placed with the open end in the headspace of a closed test system containing aqueous test medium. The test chemical served as the dominating partitioning donor for establishing and maintaining maximum exposure levels in the headspace and aqueous solution, without direct contact between the donor and the test medium. The HS-PD method was cross validated against passive dosing with a saturated silicone elastomer, using headspace gas chromatography as analytical instrument and saturated vapors as reference. The HS-PD method was then applied to control the exposure in algal growth inhibition tests with the green algae Raphidocelis subcapitata. The model chemicals were C9-C14 n-alkanes and the cyclic volatile methyl siloxanes octamethyltetracyclosiloxane (D4) and decamethylpentacyclosiloxane (D5). Growth rate inhibition at the solubility limit was 100% for C9-C13 n-alkanes and 53 ±â€¯31% (95% CI) for tetradecane. A moderate inhibition of 11 ±â€¯4% (95% CI) was observed for D4, whereas no inhibition was observed for D5. The present study introduces an effective method for aquatic toxicity testing of a difficult-to-test group of chemicals and provides an improved experimental basis for investigating toxicity cut-offs.

Deciphering mechanisms of malathion toxicity under pulse exposure of the freshwater cladoceran Daphnia magna.

The organophosphate pesticide (OP) malathion is highly toxic to freshwater invertebrates, including the cladoceran Daphnia magna, a widely used test organism in ecotoxicology. To assess whether toxic effects of malathion are driven primarily by exposure concentration or exposure duration, D. magna was pulse exposed to equivalent integrated doses (duration × concentration): 3 h × 16 µg/L, 24 h × 2 µg/L, and 48 h × 1 µg/L. After recovery periods of 3 h, 24 h, and 48 h, the toxicity of malathion on different biological levels in D. magna was examined by analyzing the following endpoints: survival and immobilization; enzyme activities of acetylcholinesterase (AChE), carboxylesterase (CbE), and glutathione S-transferase (GST); and AChE gene expression. The results showed no difference in survival among equivalent integrated doses. Adverse sublethal effects were driven by exposure concentration rather than pulse duration. Specifically, short pulse exposure to a high concentration of malathion resulted in more immobilized daphnids, lower AChE and CbE activities, and a higher transcript level of AChE gene compared with long pulse exposure to low concentration. The expression of the AChE gene was up-regulated, indicating a compensatory mechanism to cope with enzyme inhibition. The study shows the need for obtaining a better understanding of the processes underlying toxicity under realistic exposure scenarios, so this can be taken into account in environmental risk assessment of pesticides.