Cerium oxide nanoparticles (nCeO2) exert minimal adverse effects on microbial communities in soils with and without biosolids amendment.

Increased use of nano-cerium oxide (nCeO2) in an array of industrial applications has raised environmental concerns due to potential increased loadings to the soil environment. This research investigated the potential adverse effects of nCeO2 (10-30 nm) on the soil microbial community in two exposure scenarios: direct application to soil, and indirect application to soil through chemical spiking of biosolids, followed by mixing into soil. Total Ce in test soils without, and with biosolids amendment, ranged from 44 to 770, and 73 to 664 mg Ce kg-1 soil, respectively. In order to help distinguish whether observed effects were elicited by the solid-phase colloids or the activity of dissolved Ce, a soluble Ce salt (Ce (NO3)3) treatment was included in select assays. A suite of tests was used to investigate effects on critical processes: microbial growth (heterotrophic plate count), microbial activity (organic matter (OM) decomposition, enzyme activity and, nitrification) and diversity (structural and functional). Although results showed significant inhibition on microbial growth in soil without biosolids amendment at ≥ 156 mg Ce kg-1 soil by week 5, these results were inconsistent and non-significant thereafter. In general, nCeO2 showed no evidence of consistent adverse effects on OM decomposition, nitrification, soil enzyme activities and functional diversity. Leucine aminopeptidase showed significant (p< 0.05) stimulatory effects over time at ≥ 44 mg Ce kg-1 in soils without biosolids, which was not observed in soils with biosolids amendment. The lack of inhibitory effects of nCeO2 may be attributed to its low solubility; Ce in soil extracts (0.01 M CaCl2) were all below detection (< 0.003 mg kg-1) in the nCeO2-spiked soils, but detectable in the Ce (NO3)3 samples. In contrast, soluble Ce at 359 mg Ce kg-1 showed a significant reduction in OM decomposition and effects on microbial genomic diversity based on the 16S rDNA data in soils with and without biosolids amendment (359 and 690 mg Ce kg-1). The nCeO2 behaviour and effects information described herein are expected to help fulfill data gaps for the characterization of this priority nanomaterial.

Soil invertebrate toxicity and bioaccumulation of nano copper oxide and copper sulphate in soils, with and without biosolids amendment.

The fate, toxicity and bioaccumulation of copper oxide nanoparticles (nCuO) was investigated in soil, with and without biosolids amendment, through chronic exposures using the earthworm, Eisenia andrei, and the collembolan, Folsomia candida. The effects of copper sulphate (CuSO4) were included so as to compare the behavior of nCuO to a readily soluble counterpart. The fate of nCuO was evaluated through characterization of dissolved and nano-particulate fractions (via single particle ICP-MS) as well as extractable Cu2+ throughout the duration of select tests. Neither Cu form was particularly toxic to F. candida, but effects on E. andrei reproduction were significant in all treatments (IC50 range: 98 - 149 mg Cu kg-1 dry soil). There were no significant differences in toxicity between the Cu forms, nor in extractable Cu2+ activities, indicative that particle dissolution within the soil and, subsequent activity of Cu2+ was likely the primary mode of toxicity in the nCuO exposures. The presence of biosolids did not significantly alter toxicity of nCuO, but did affect Cu2+ activity over time. Bioaccumulation of total Cu in E. andrei when exposed to nCuO (kinetic bioaccumulation factor (BAFk): 0.80 with biosolids and 0.81 without) was lower than exposure to CuSO4 (BAFk: 2.31 with biosolids and 1.12 without). Enhanced dark-field hyperspectral imaging showed accumulation of nCuO along the epidermis and gut of E. andrei, with trace amounts observed in muscle and chloragogenous tissue, providing evidence of nCuO translocation within the organism. The present study demonstrates that the current risk assessment approach for trace metals in the environment, based on substance solubility and bioavailability of the dissolved free ion, are applicable for nCuO exposure to soil invertebrates, but that the rate of particle dissolution in different soil environments is an important factor for consideration.

Lethal and Sublethal Toxicity of Thiamethoxam and Clothianidin Commercial Formulations to Soil Invertebrates in a Natural Soil.

The use of neonicotinoids in agriculture is a critical environmental protection issue. Although there has been considerable research on pollinator exposure and aquatic toxicological effects, few studies have investigated the chronic impacts on soil-dwelling species. Given the application of neonicotinoids into soil systems, there is the potential for risk to soil invertebrates. The toxicity of 2 commercial formulations containing the active ingredients (a.i.) thiamethoxam (Actara® 240SC) or clothianidin (Titan™) was investigated using 3 soil invertebrate species: Oppia nitens, Eisenia andrei, and Folsomia candida. No adverse effects were observed for O. nitens at the highest tested concentrations (≥92 mg a.i./kg dry soil) after a 28-d exposure. Exposure to clothianidin resulted in a 28-d median inhibitory concentration (IC50) of 0.069 (95% confidence limits: 0.039-0.12) mg/kg dry soil for F. candida, and a 56-d IC50 of 0.26 (0.22-3.2) mg a.i./kg dry soil for E. andrei. Exposure to thiamethoxam was less toxic, with IC50s of 0.36 (0.19-0.66) and 3.0 (2.2-4.0) mg a.i./kg dry soil for F. candida and E. andrei reproduction, respectively. The observed toxicity for F. candida adult survival and reproduction and for E. andrei reproduction occurred at environmentally relevant concentrations. However, because clothianidin is a degradation product of thiamethoxam, and detection of clothianidin rose to levels of concern in the thiamethoxam-amended soils over time, the observed toxicity may be partly attributed to the presence of clothianidin. Environ Toxicol Chem 2019;38:2111-2120. © 2019 Crown in the right of Canada. Published by Wiley Periodicals Inc. on behalf of SETAC.

Perfluorooctane sulfonate in surface soils: Effects on reproduction in the collembolan, Folsomia candida, and the oribatid mite, Oppia nitens.

Perfluorooctane sulfonate (PFOS) is a persistent organic pollutant, which has been detected at significant concentrations in soils at sites used for fire-fighting training operations. Recent ecotoxicological research has mainly focused on earthworms to assess the toxicity of PFOS in soil. However, the inclusion of other soil taxonomic groups allow for a more holistic estimate of contaminant risk, including the derivation of more comprehensive soil quality guidelines. The present study assessed the toxicity of PFOS using the collembolan, Folsomia candida, and the oribatid mite, Oppia nitens, in two types of soil: a coarse-textured sandy loam (VSL) and fine-textured clay loam (NRS). As a standard O. nitens reproduction test is being formalized, the results of the study were also used to compare sensitivity across test species. Effects were soil dependent, with test species being 2-4 times more susceptible to PFOS in VSL, relative to NRS, likely due to differences in organic matter and clay content. Oppia nitens was significantly more sensitive to PFOS, regardless of soil type, in comparison to F. candida. The IC50s for reproduction for O. nitens were 23 mg kg-1 (95% confidence interval: 17-32 mg kg-1) in the VSL and 95 mg kg-1 (69-134 mg kg-1) in the NRS, and for F. candida were 94 mg kg-1 (72-122 mg kg-1) in the VSL and 233 mg kg-1 (177-306 mg kg-1) in the NRS. The present study demonstrates the application and inclusion of the oribatid mite, O. nitens, for the risk assessment of contaminants in soil.

Ecotoxicity of boric acid in standard laboratory tests with plants and soil organisms.

To verify the continuous sensitivity of ecotoxicological tests (mainly the test organisms), reference substances with known toxicity are regularly tested. Ideally, this substance(s) would lack specificity in its mode action, be bioavailable and readily attainable with cost-effective means of chemical characterization. Boric acid has satisfied these criteria, but has most recently been characterized as a substance of very high concern, due to reproductive effects in humans, thus limiting its recommendation as an ideal reference toxicant. However, there is probably no other chemical for which ecotoxicity in soil has been so intensively studied; an extensive literature review yielded lethal (including avoidance) and sublethal data for 38 taxa. The ecotoxicity data were evaluated using species sensitivity distributions, collectively across all taxa, and separately according to species type, endpoints, soil type and duration. The lack of specificity in the mode of action yielded broad toxicity among soil taxa and soil types, and provided a collective approach to assessing species sensitivity, while taking into consideration differences in test methodologies and exposure durations. Toxicity was species-specific with Folsomia candida and enchytraied species demonstrating the most sensitivity; among plants, the following trend occurred: dicotyledonous (more sensitive) ≫ monocotyledonous ≫ gymnosperm species. Sensitivity was also time and endpoint specific, with endpoints such as lethality and avoidance being less sensitive than reproduction effects. Furthermore, given the breadth of data and toxicity demonstrated by boric acid, lessons learned from its evaluation are discussed to recommend the properties required by an ideal reference substance for the soil compartment.

Validation of a new standardized test method for the freshwater amphipod Hyalella azteca: Determining the chronic effects of silver in sediment.

Environment Canada has developed a new 42-d sediment toxicity test method that includes a reproduction test endpoint with the freshwater amphipod Hyalella azteca. Because of concerns that existing standard methodologies, whereby adults are transferred to a water-only exposure before release of their first brood at day 28, will lead to internal contaminant depuration and loss of sensitivity, the Environment Canada methodology conducts the entire exposure in sediment. To demonstrate applicability of the method for assessing the toxicity of chemical-spiked sediment, H. azteca were exposed for 42 d to sediment amended with silver nitrate (AgNO3 ). Mortality was significantly higher at the highest sediment concentration of Ag (2088 mg/kg dry wt); however, there was no significant reduction in biomass or reproduction as a result of Ag exposure despite significant bioaccumulation. Based on Ag measurements and speciation modeling, the principle route of Ag exposure was likely through the ingestion of complexed colloidal or particulate Ag. The techniques used to recover young amphipods from sediment were critical, and although this effort can be labor intensive (20-45 min/replicate), the technicians demonstrated 91% recovery in blind trials. For the first time, Environment Canada will require laboratories to report their recovery proficiency for the 42-d test-without this information, data will not be accepted. Overall, the reproduction test will be more applicable when only a few chemical concentrations need to be evaluated in laboratory-amended sediments or for field-collected contaminated site assessments (i.e., contaminated site vs reference site comparisons). Environ Toxicol Chem 2016;35:2430-2438. © 2016 SETAC.

Review of laboratory-based terrestrial bioaccumulation assessment approaches for organic chemicals: Current status and future possibilities.

In the last decade, interest has been renewed in approaches for the assessment of the bioaccumulation potential of chemicals, principally driven by the need to evaluate large numbers of chemicals as part of new chemical legislation, while reducing vertebrate test organism use called for in animal welfare legislation. This renewed interest has inspired research activities and advances in bioaccumulation science for neutral organic chemicals in aquatic environments. In January 2013, ILSI Health and Environmental Sciences Institute convened experts to identify the state of the science and existing shortcomings in terrestrial bioaccumulation assessment of neutral organic chemicals. Potential modifications to existing laboratory methods were identified, including areas in which new laboratory approaches or test methods could be developed to address terrestrial bioaccumulation. The utility of "non-ecotoxicity" data (e.g., mammalian laboratory data) was also discussed. The highlights of the workshop discussions are presented along with potential modifications in laboratory approaches and new test guidelines that could be used for assessing the bioaccumulation of chemicals in terrestrial organisms.

Estimation of the bioaccumulation potential of a nonchlorinated bisphenol and an ionogenic xanthene dye to Eisenia andrei in field-collected soils, in conjunction with predictive in silico profiling.

In silico-based model predictions, originating from structural and mechanistic (e.g., transport, bioavailability, reactivity, and binding potential) profiling, were compared against laboratory-derived data to estimate the bioaccumulation potential in earthworms of 2 organic substances (1 neutral, 1 ionogenic) known to primarily partition to soil. Two compounds representative of specific classes of chemicals were evaluated: a nonchlorinated bisphenol containing an -OH group (4,4'-methylenebis[2,6-di-tert-butylphenol] [Binox]), and an ionogenic xanthene dye (2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3',6'-dihydroxy-, disodium salt [Phloxine B]). Soil bioaccumulation studies were conducted using Eisenia andrei and 2 field-collected soils (a clay loam and a sandy soil). In general, the in silico structural and mechanistic profiling was consistent with the observed soil bioaccumulation tests. Binox did not bioaccumulate to a significant extent in E. andrei in either soil type; however, Phloxine B not only accumulated within tissue, but was not depurated from the earthworms during the course of the elimination phase. Structural and mechanistic profiling demonstrated the binding and reactivity potential of Phloxine B; this would not be accounted for using traditional bioaccumulation metrics, which are founded on passive-based diffusion mechanisms. This illustrates the importance of profiling for reactive ionogenic substances; even limited bioavailability combined with reactivity can result in exposures to a hazardous substance not predictable by traditional in silico modeling methods.

Ecotoxicity of xanthene dyes and a non-chlorinated bisphenol in soil.

Soil eco-toxicity testing was conducted in support of Canada's Chemical Management Plan (CMP) to fill data gaps for organic chemicals known to primarily partition to soil, and of which the persistence and inherent toxicity are uncertain. Two compounds representative of specific classes of chemicals: non-chlorinated bisphenols containing an -OH group (4,4'-methylenebis(2,6-di-tert-butylphenol (Binox)) and xanthene dyes (2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3',6'-dihydroxy-, disodium salt (Phloxine B), 2',4',5',7'-tetrabromofluorescein (TBF), 4',5'-dibromofluorescein (DBF), and 4,5,6,7-tetrachlorofluorescein (TCF)) were evaluated. The effect of these substances on plant growth (Elymus lanceolatus and Trifolium pratense) and soil invertebrate survival and reproduction (Folsomia candida and Eisenia andrei) were assessed using a field-collected sandy soil. Binox was persistent throughout testing (up to 63 d) with an average recovery of 77±2.9% at test end. Binox was not toxic to plants (IC50s>1076 mg kg(-1)) or E. andrei (IC50s>2651 mg kg(-1)); however, a significant reduction in F. candida adult survival and reproduction (IC50=89 (44-149) mg kg(-1)) was evident. Phloxine B was also persistent throughout testing, with an average recovery of 82±3.0% at test end. Phloxine B was significantly more toxic than Binox, with significant reductions in plant root growth (IC50s ≥ 11 mg kg(-1)) and invertebrate reproduction (IC50s ≥ 22 mg kg(-1)). DBF toxicity was not significantly different from that of Phloxine B for plant root growth (IC50s ≥ 30 mg kg(-1)), but was significantly less toxic for shoot growth (IC50s ≥ 1758 mg kg(-1)), and invertebrate adult survival (IC50s ≥ 2291 mg kg(-1)) and reproduction (IC50s ≥ 451 mg kg(-1)). A comparison between all four xanthene dyes was completed using F. candida, with the degree of toxicity in the order of Phloxine B ≥ TBF∼DBF>TCF. The results from these studies will contribute to data gaps for poorly understood chemicals (and chemical groupings) under review for environmental risk assessments, and will aid in the validation of model predictions used to characterize the fate and effects of these substances in soil environments.

Evaluation of a new battery of toxicity tests for boreal forest soils: assessment of the impact of hydrocarbons and salts.

The ability to assess the toxic potential of soil contamination within boreal regions is currently limited to test species representative of arable lands. This study evaluated the use of six boreal plant species (Pinus banksiana, Picea glauca, Picea mariana, Populus tremuloides, Calamagrostis Canadensis, and Solidago canadensis) and four invertebrate species (Dendrodrilus rubidus, Folsomia nivalis, Proisotoma minuta, and Oppia nitens) and compared their performance to a suite of standard agronomic soil test species using site soils impacted by petroleum hydrocarbon (PHC) and salt contamination. To maintain horizon-specific differences, individual soil horizons were collected from impacted sites and relayered within the test vessels. Use of the boreal species was directly applicable to the assessment of the contaminated forest soils and, in the case of the hydrocarbon-impacted soil, demonstrated greater overall sensitivity (25th percentile of estimated species sensitivity distribution [ESSD25] = 5.6% contamination: 10,600 mg/kg fraction 3 [F3; equivalent hydrocarbon range of >C16 to C34] Of/Oh horizon, and 270 mg/kg F3 Ahg horizon) relative to the standard test species (ESSD25 = 23% contamination: 44,000 mg/kg F3 Of/Oh horizon, and 1,100 mg/kg F3 Ahg horizon). For salinity, there was no difference between boreal and standard species with a combined ESSD25 = 2.3%, equating to 0.24 and 0.25 dS/m for the Ah and Ck horizons. The unequal distribution of soil invertebrates within the layered test vessels can confound test results and the interpretation of the toxic potential of a site. The use of test species relevant to boreal eco-zones strengthens the applicability of the data in support of realistic ecological risk assessments applicable to the boreal regions.

Ecotoxicity of siloxane D5 in soil.

Decamethylcyclopentasiloxane (D5) is a cyclic volatile methyl siloxane (cVMS) commonly found in commercially available products. D5 is expected to enter the terrestrial environment through the deposit of biosolids from sewage treatment plants onto agricultural fields for nutrient enrichment. Little to no information currently exists as to the risks of D5 to the terrestrial environment. In order to evaluate the potential risk to terrestrial organisms, the toxicity of a D5 contaminated biosolid in an agricultural soil was assessed with a battery of standardized soil toxicity tests. D5 was spiked into a surrogate biosolid and then mixed with a sandy loam soil to create test concentrations ranging from 0 to 4074 mg kg(-1). Plant (Hordeum vulgare (barley) and Trifolium pratense (red clover)) and soil invertebrates (Eisenia andrei (earthworm) and Folsomia candida (springtail)) toxicity tests were completed to assess for lethal and sub-lethal effects. Plant testing evaluated the effects on seedling emergence, shoot and root length, and shoot and root dry mass. Invertebrate test endpoints included adult lethality, juvenile production, and individual juvenile dry mass (earthworms only). Soil samples were collected over time to confirm test concentrations and evaluate the loss of chemical over the duration of a test. The toxicity of the D5 was species and endpoint dependent, such that no significant adverse effects were observed for T. pratense or E. andrei test endpoints, however, toxicity was observed for H. vulgare plant growth and F. candida survival and reproduction. Chemical losses of up to 50% were observed throughout the tests, most significantly at high concentrations.

Identification and application of AFLP-derived genetic markers for quantitative PCR-based tracking of Bacillus and Paenibacillus spp. released in soil.

In this study, we show that noncoding sequences from amplified fragment length polymorphisms (AFLPs) can provide robust and sensitive genetic markers suitable for PCR-based discrimination of closely related strains of Bacillus and Paenibacillus, and quantitative PCR (qPCR)-based tracking of the strains in complex natural systems like soil. Quantitative PCR was accurate in the approximately 1 x 10(9) to approximately 1 x 10(4) colony forming units (CFU)/g soil range. The detection limit was improved to approximately 1 x 10(2) CFU/g when amplicons were analyzed by gel electrophoresis. Studies with laboratory-contained intact soil-core microcosms indicated that environmental persistence trends vary among different strains. For example, Bacillus circulans ATCC 9500, Bacillus amyloliquefaciens DSL 13563-0, Bacillus licheniformis ATCC 12713, Paenibacillus polymyxa NRRL B-4317, and 3 Bacillus subtilisstrains (ATCC 6051A, ATCC 55405, and NRRL B-941) died down to below the 1 x 10(2) CFU/g detection limit by days 28-105. In contrast, over a 105-day period, B. licheniformis ATCC 55406, Bacillus megaterium NRRL B-14308, and P. polymyxa strains ATCC 55407 and DSL 13540-4 died down but persisted at levels just above the detection limit, whereas Bacillus thuringiensis ATCC 13367 experienced a less than 10-fold decrease in cell numbers.