Richard P. Scroggins

Oribatid mites in soil toxicity testing-the use of Oppia nitens (C.L. Koch) as a new test species.

Few soil invertebrate species are available for the toxic assessment of soils from boreal or other northern ecozones, yet these soils cover the majority of Canadas landmass as well as significant portions of Eurasia. Oppia nitens (C.L. Koch) is an herbivorous and fungivorous oribatid mite found in soil throughout Holarctic regions, including Canada. Soil tests using O. nitens were performed using 15 different forest soil types and horizons to investigate test variability in adult survival and reproduction. Adult survival (86.1 +/- 1.1%) was consistent across soil types, with a coefficient of variation (CV) of 15%. However, reproduction varied significantly, ranging from 2.9 (+/-1.1) to 86.2 (+/-11.7) individuals, with a corresponding CV of 118 and 30%, respectively. Of the soil factors assessed (NH(3), NO(3), pH, phosphorus [P], organic matter content (OM), carbon:nitrogen (C:N), sand, silt, clay, and sodium adsorption ratio), soil organic matter (OM) explained 68% of the variation observed for reproduction. Increasing the OM using Sphagnum sp. peat moss resulted in optimal reproduction at 7% OM (8% peat content) with the lowest variability (CV of 20%). When assessing the toxicity of a reference chemical, boric acid, the effect of peat amendment reduced lethality to adults with no observable difference on reproduction. The use an age-synchronized culture reduced the test variability for reproduction relative to the use of unsynchronized cultures. Oppia nitens is a good candidate species for a standardized test design, with adult survival easily assessed in a relatively simple design. A long-term reproduction test with O. nitens will require the use of a synchronized population and, on occasion, OM amendment when testing soils with low organic matter content.

A comparison of the effects of silver nanoparticles and silver nitrate on a suite of soil dwelling organisms in two field soils

Abstract Nanomaterials are increasingly used in a wide range of products, leading to growing concern of their environmental fate. In order to understand the fate and effects of silver nanoparticles in the soil environment, a suite of toxicity tests including: plant growth with Elymus lanceolatus (northern wheatgrass) and Trifolium pratense (red clover); collembolan survival and reproduction (Folsomia candida); and earthworm avoidance, survival and reproduction (Eisenia andrei) was conducted. The effect of silver nanoparticles (AgNP) was compared with the effect of ionic silver (as AgNO3) in two agricultural field soils (a sandy loam and a silt loam). Lethal (LC50) or sub lethal (IC50) effect levels are presented for all endpoints and demonstrate that in most cases AgNO3 (i.e. ionic silver) was found to be more toxic than the AgNP across test species. The difference in effects observed between the two forms of silver varied based on test species, endpoint and soil type. In tests that were conducted across different soil types, organisms in the sandier soil had a greater response to the Ag (ionic and nano) than those in soil with a high silt content. Earthworms (avoidance behavior and reproduction) were the most sensitive to both AgNP and AgNO3, while plant emergence was the least sensitive endpoint to both forms of Ag. The use of a test battery approach using natural field soils demonstrates the need to better quantify the dissolution and transformation products of nanomaterials in order to understand the fate and effects of these materials in the soil environment.

Effects of metal‐contaminated forest soils from the Canadian shield to terrestrial organisms

The effects of elevated metal concentrations in forest soils on terrestrial organisms were investigated by determining the toxicity of six site soils from northern Ontario and Quebec, Canada, using a battery of terrestrial toxicity tests. Soils were collected from three sites on each of two transects established downwind of nickel (Sudbury, ON, Canada) and copper (Rouyn-Noranda, PQ, Canada) smelting operations. Site soils were diluted to determine if toxicity estimates for the most-contaminated site soils could be quantified as a percent of site soil. Rouyn-Noranda soils were toxic following acute exposure (14 d) to plants, but not to invertebrates (7 d for collembola and 14 d for earthworms). However, Rouyn-Noranda soils were toxic to all species following chronic exposure (21, 35, and 63 d for plants, collembola, and earthworms, respectively). The toxicity of the Rouyn-Noranda site soils did not correspond to the gradient of metal concentrations in soil. Metal-contaminated Sudbury soils were toxic to plants but not to invertebrates, following acute exposure. Chronic exposure to Sudbury soils caused adverse effects to plant growth and invertebrate survival and reproduction. The toxicity of Sudbury soils corresponded to the metal concentration gradient, with one exception: The reference soil collected in October was toxic to collembola following acute and chronic exposure. This study evaluated the applicability of the new Environment Canada terrestrial toxicity test methods, developed using agricultural soils, to forest soils and also provided useful data to assess the ecological risk associated with mixtures of metals in soil.

Development of Suitable Test Methods for Evaluating the Toxicity of Contaminated Soils to a Battery of Plant Species Relevant to Soil Environments in Canada

Environment Canada and the Canadian Association of Petroleum Producers (CAPP) have embarked on a five year program to develop, standardize, and validate a battery of soil toxicity tests which can be used to assess the relative toxicity of contaminants in soils to terrestrial organisms. These tests must be applicable to soil conditions typically found in Canadian environments and the test species must be representative of the species of soil invertebrates or plants inhabiting soil ecosystems in Canada. Two of the tests being considered for development are a seed germination test and a seedling growth test using whole soils. The battery of plant species that is being considered for testing includes thirteen species representing six dicotyledonous families and 17 species representing two monocotyledonous families. The species of plants with one exception (Arabidopsis sp.) are representative of both above- and below-ground crop plants grown in agricultural lands across Canada, as well as non-crop plants that are also widely distributed in Canada. One of the objectives of the project is to screen 30 plant species and select ten species that are suitable for toxicity assessment of contaminated soils. In addition to selection of the most appropriate test species, research has focused on test method development. Comparison of toxicity test endpoints from reference tests permitted the assessment of the variation associated with each endpoint (root length, stem length, wet and dry mass of stem and root) for each species tested to date. The tests were performed using boric acid as the reference toxicant and an artificial soil and a field-collected reference soil as control soils. The methods and results from toxicity tests have been summarized and presented with recommendations regarding test species, methods, endpoints, and conditions.

Extracting Metallic Nanoparticles from Soils for Quantitative Analysis: Method Development Using Engineered Silver Nanoparticles and SP-ICP-MS

The lack of an efficient and standardized method to disperse soil particles and quantitatively subsample the nanoparticulate fraction for characterization analyses is hindering progress in assessing the fate and toxicity of metallic engineered nanomaterials in the soil environment. This study investigates various soil extraction and extract preparation techniques for their ability to remove nanoparticulate Ag from a field soil amended with biosolids contaminated with engineered silver nanoparticles (AgNPs), while presenting a suitable suspension for quantitative single-particle inductively coupled plasma mass spectroscopy (SP-ICP-MS) analysis. Extraction parameters investigated included reagent type (water, NaNO3, KNO3, tetrasodium pyrophosphate (TSPP), tetramethylammonium hydroxide (TMAH)), soil-to-reagent ratio, homogenization techniques as well as procedures commonly used to separate nanoparticles from larger colloids prior to analysis (filtration, centrifugation, and sedimentation). We assessed the efficacy of the extraction procedure by testing for the occurrence of potential procedural artifacts (dissolution, agglomeration) using a dissolved/particulate Ag mass ratio and by monitoring the amount of Ag mass in discrete particles. The optimal method employed 2.5 mM TSPP used in a 1:100 (m/v) soil-to-reagent ratio, with ultrasonication to enhance particle dispersion and sedimentation to settle out the micrometer-sized particles. A spiked-sample recovery analysis shows that 96% ± 2% of the total Ag mass added as engineered AgNP is recovered, which includes the recovery of 84.1% of the particles added, while particle recovery in a spiked method blank is ∼100%, indicating that both the extraction and settling procedure have a minimal effect on driving transformation processes. A soil dilution experiment showed that the method extracted a consistent proportion of nanoparticulate Ag (9.2% ± 1.4% of the total Ag) in samples containing 100%, 50%, 25%, and 10% portions of the AgNP-contaminated test soil. The nanoparticulate Ag extracted by this method represents the upper limit of the potentially dispersible nanoparticulate fraction, thus providing a benchmark with which to make quantitative comparisons, while presenting a suspension suitable for a myriad of other characterization analyses.

Ion exchange technique (IET) to characterise Ag+ exposure in soil extracts contaminated with engineered silver nanoparticles

Environmental context Biosolid-amended soils are likely sinks for manufactured silver nanoparticles, the environmental toxicity of which is believed to be related to the release and accumulation of Ag+ ions. This study demonstrates how an ion exchange technique can be applied to soil extracts to provide Ag+ measurements at low, environmentally relevant levels. The technique is a valuable addition to existing analytical methods for tracking the behaviour of Ag nanoparticles and Ag+ ions in the terrestrial environment. Abstract The lack of silver speciation exposure data in toxicity studies investigating the effects of manufactured silver nanoparticles (AgNPs) in natural soil media limits the ability to discern nano-specific effects from effects of the toxic Ag+ form, which may be released from the manufactured AgNPs contained in wastewater, biosolids or soil environment. Using samples containing Ag+ or mixtures of Ag+ and AgNPs, ranging in total Ag concentrations of 10–5 to 10–9 M, and prepared in de-ionised water and filtered soil extracts, the validity of the ion exchange technique (IET) to quantify Ag+ was investigated by comparing measurements to those of an Ag+ ion selective electrode (ISE) and to the dissolved fraction from single particle inductively coupled plasma–mass spectrometry (SP-ICP-MS) analysis (SP-dissolved). When analysing samples in the filtered soil extract, IET and ISE gave comparable results down to 10–7 M, below which Ag+ activities were below the ISE detection limit. For water samples, SP-dissolved values were generally comparable or slightly greater (on average 65%) compared with IET-Ag+ at all concentrations. The high bias was likely due to inclusion of unresolved particles below the SP-ICP detection limit of 19nm. However, when analysing samples in the soil extract, SP-dissolved values were on average eight-fold greater than IET-Ag+, highlighting the effect that natural colloidal and dissolved soil constituents have on complexing Ag+, as well as the lack of specificity of the SP-dissolved analysis for the Ag+ species. IET is shown here to be a valid procedure to quantify Ag+ activity in soil extracts, and while the study highlights the limitations of using the SP-dissolved fraction to estimate this biologically relevant Ag fraction, it shows that combined, IET and SP-ICP-MS provide a valuable approach for investigating the behaviour of manufactured AgNPs in different matrixes.

Phytotoxkit: a critical look at a rapid assessment tool.

Terrestrial plant toxicity testing contributes critical information to many site risk assessments, but standardized tests can be labor-intensive, use large amounts of soil, and employ long test durations. The Phytotoxkit (MicroBioTests, Environmental Bio-Detection Products) minimizes the time and cost associated with terrestrial plant testing with a unique test setup, a shorter test duration, and less soil. However, the sensitivity of the test remains an open question. In this research, the Phytotoxkit and the standardized Environment Canada terrestrial plant toxicity test (definitive test) are compared using a parallel testing approach. Three different scenarios were examined: a multiconcentration test, in which an inhibiting concentration (ICp) was derived from chemically amended soils; a soil remediation test, in which plant growth in a remediated soil was compared to the original contaminated soil; and a site soil test, in which plant growth in a contaminated soil was compared to a reference soil. The contaminants tested were boric acid, Cr(VI) with cyclodextrin as a remediation agent, and petroleum hydrocarbons. Trifolium pratense (red clover) was used in the first and second scenarios, and six different plant species were used in the third scenario. In the first scenario, the Phytotoxkit results compared well with the definitive test results after 5 and 7 d of exposure. In the second scenario, the Phytotoxkit results agreed with the definitive test when evaluating the effectiveness of remediation. In the third scenario, the Phytotoxkit results were often not in agreement with the results from the definitive test. The reduced sensitivity of the Phytotoxkit in the third scenario may be driven by test unit design, as plant roots are separated from soil by filter paper.

Sublethal Toxicity Testing of Canadian Metal Mining Effluents: National Trends and Site-Specific Uses

ABSTRACT As part of the Canadian Environmental Effects Monitoring program under the National Metal Mining Effluent Regulation, there is a requirement to conduct sublethal toxicity tests twice per year for the first three years. These first three years (2003 to 2005) were considered a period of initial monitoring and resulted in test endpoints for each of the required standardized methods on a fish, an aquatic plant, an invertebrate, and an algal species. On a national level (based on 1648 valid results), the test from most to least sensitive was: the inhibition of reproduction with Ceriodaphnia dubia, the growth inhibition (frond number) with Lemna minor, the inhibition of cell yield with Pseudokirchneriella subcapitata, the growth inhibition (dry weight) with Lemna minor, the growth inhibition of fathead minnows, and the effect on embryo viability with rainbow trout. This sensitivity ranking changed when tests were further evaluated on a geographical region and mine-type basis (e.g., base metal, precious metal, uranium, iron ore). Site-specific examples show how sublethal toxicity data are being used to track changes in effluent quality, choosing a final discharge point, monitoring multiple discharges to the same watercourse, and to identify study design weaknesses by comparing laboratory results to field survey conclusions.

The Lab-to-Field (LTF) Rating Scheme: A New Method of Investigating the Relationships between Laboratory Sublethal Toxicity Tests and Field Measurements in Environmental Effects Monitoring Studies

ABSTRACT The Lab-to-Field (LTF) rating scheme is a straightforward method of relating the results of effluent toxicity tests to the field survey measurements and has proven to be a useful tool for interpretation of Environmental Effects Monitoring studies for pulp and paper mills in the province of Ontario. The LTF method uses the same five-level scale (level 1 for no or low response to level 5 for severe response) for rating the toxicity and field survey results. Regression analysis of LTF scores has revealed that the relationship between the Ceriodaphnia reproduction test and benthic invertebrate field survey measurements was significant (p < 0.001, r = 0.79). However, there were not sufficient data to determine if this can be used as a predictive tool. Nonetheless, Ceriodaphnia-to-benthic survey, Selenastrum-to-benthic survey and fathead-to-fish survey relationships were qualitatively rated strong or moderately strong in 94%, 75%, and 60% of the 16 studies, respectively. The LTF rating scheme would benefit from the use of a more sensitive species or life stage of fish to strengthen the sublethal test-to-fish survey relationship. Further validation of the lab-to-field relationship could be accomplished through the synoptic collection of effluent samples for sublethal tests and field measurements.

The ecotoxicity of zinc and zinc‐containing substances in soil with consideration of metal‐moiety approaches and organometal complexes

Within Canada, screening-level assessments for chemical substances are required to determine whether the substances pose a risk to human health and/or the environment, and as appropriate, risk management strategies. In response to the volume of metal and metal-containing substances, process efficiencies were introduced using a metal-moiety approach, whereby substances that contain a common metal moiety are assessed simultaneously as a group, with the moiety of concern consisting of the metal ion. However, for certain subgroups, such as organometals or organic metal salts, the organic moiety or parent substance may be of concern, rather than simply the metal ion. To further investigate the need for such additional consideration, certain substances were evaluated: zinc (Zn)-containing inorganic (Zn chloride [ZnCl2] and Zn oxide) and organic (organometal: Zn diethyldithiocarbamate [Zn(DDC)2 ] and organic metal salts (Zn stearate [ZnSt] and 4-chloro-2-nitrobenzenediazonium tetrachlorozincate [BCNZ]). The toxicity of the substances were assessed using plant (Trifolium pratense and Elymus lanceolatus) and soil invertebrate (Folsomia candida and Eisenia andrei) tests in a sandy soil. Effect measures were determined based on total metal and total parent analyses (for organic substances). In general, the inorganic Zn substances were less toxic than the organometals and organic metal salts, with 50% effective concentrations ranging from 11 to >5194 mg Zn kg-1 dry soil. The data demonstrate the necessity for alternate approaches in the assessment of organo-metal complexes, with the organic moieties or parent substances warranting consideration rather than the metal ion alone. In this instance, the organometals and organic metal salts were significantly more toxic than other test substances despite their low total Zn content. Environ Toxicol Chem 2017;36:3324-3332.