Assessment of Contaminated Soil in the Canadian Boreal Forest using Standardized Toxicity tests. Mary Moody, SRC Rick Scroggins, Environment Canada

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Assessment of Contaminated Soil in the Canadian Boreal Forest using Standardized Toxicity tests Mary Moody, SRC Rick Scroggins, Environment Canada

Outline 1. Methods Plants Invertebrates Soil collection and handling 2. Performance tests 3. Case study of hydrocarbon impacted site http://www.borealforest.org/world/world_overview.htm

Project Objectives Develop standardized biological test methods to measure effects of contaminants (e.g., brine, PHC) in Canadian boreal and northern (taiga) soils and wetlands Use ecologically-relevant terrestrial and wetland species (single-species and microbial) Test a variety of soils of the boreal and taiga eco-zones Develop technical guidance on the collection, handling, and preparation of contaminated soils for biological testing in support of site-specific risk assessments Develop tools useful for contaminated land risk assessment and management

Method development process Create list of potential species that are ecologically relevant to the boreal Acquire or collect seeds or invertebrates, bring to germination or into culture in the lab Perform tests of growth, survival or reproduction in a wide variety of soils performance testing Test impacted soils diluted in a matching reference soil Environment Canada validates the method, publishes

Contaminated Soil Sampling Guidance Environment Canada EPS 1/RM/53 Supportive of site-specific risk assessments and soil remediation Single-species and microbial assessments Universal procedures and statistically derived sampling procedures Specific procedures for problematic contamination (e.g., unstable compounds, volatiles) Broad-range of Canada s eco-zones (e.g., forests, cryosols, stoney soils, wetlands)

Boreal Test Species Species Plants Earthworm Black spruce (Picea mariana) White spruce (Picea glauca) Paper birch (Betula papyrifera) Jack pine (Pinus banksiana) Trembling aspen (Populus tremuloides) Bluejoint reedgrass (Calamagrostis canadensis) Canada goldenrod (Solidago canadensis) Dendrodrilus rubidis Source Commercial seed suppliers or locally collected Alberta Dendrobaena octaedra Springtail Proisotoma minuta Sask. Mite Oppia nitens Ontario

Wetland Plant Test Species Habitat Species Source Bebb s willow (Salix bebbiana) Marsh Cattail (Typha latifolia) Bluejoint reedgrass (Calamagrostis canadensis) Bebb s willow (Salix bebbiana) Aquatic sedge (Carex aquatilis) Fen Bog Aquatic sedge (Carex aquatilis) Tamarack (Larix laricina) Sweet gale (Myrica gale) Black spruce (Picea mariana) Bluejoint reedgrass (Calamagrostis canadensis) Bog cranberry (Vaccinium vitis-idaea) Black spruce (Picea mariana) Commercial seed suppliers or locally collected

Establish Invertebrate Species Derived from heat extractions of reference soil Ongoing culture in the lab on artificial substrate Hand-collection and sorting Culturing

Bulk soil collection sites Chernozem Gleysol Podzol Luvisol, Brunisol & Wetlands Podzol Podzol

Collection of bulk soils - aim to retain soil horizons Ah Bt Ah Bt Field horizons Plant test setup

Effects of salt-contaminated soil Roots penetrate both horizons Both shoot and root demonstrate effects of salt

Invertebrate Tests Systems Testing of Individual Horizons mite Collembolan Earthworm

Performance of plants (root length)

Performance of Dd. rubidus in 14 boreal soils Juvenile production varies with soil type but adult survival not affected Reduced reproduction in acidic soil (ph <4.5) Mean Juvenile Production

Performance of Oppia nitens in 14 boreal soils Evaluate survival and reproduction in boreal soils Develop performance data to help establish validity criteria Mean Juvenile Production OM an influential factor

Performance of P. minuta in 14 boreal soils

Site of crude oil spill in 1989 Swan Hills bog in Northern Alberta water table of 5-15 cm ph 4.6 20-30 cm peat layer over 10-20 cm Ahg horizon trees removed and straw spread to remediate 35% of surface not vegetated hydrocarbons high in F3: >C16-C34: 190,000 mg/kg Test species present: black spruce, bluejoint, paper birch, trembling aspen 2006 Collected two horizons Of/Oh and Ahg at reference and contaminated locations

Plant shoot and root endpoints (IC25) 120 Shoot Length (IC25) 70 Shoot Dry Weight (IC25) 100 60 IC25 (% dilution) 80 60 40 IC25 (% dilution) 50 40 30 20 20 10 0 black white jack pine aspen bluejoint goldenrod spruce spruce 0 black white jack pine aspen bluejoint goldenrod spruce spruce 120 Root Length (IC25) 120 Root Dry Weight (IC25) 100 100 IC25 (% dilution) 80 60 40 20 IC25 (%dilution) 80 60 40 20 0 black white jack pine aspen bluejoint goldenrod spruce spruce 0 black white jack pine aspen bluejoint goldenrod spruce spruce

Boreal invertebrate survival and juvenile production (IC25) 120 Adult Lethality (LC50) 8 Juvenile Production (IC25) LC50 (% dilution) 100 80 60 40 IC25 (% dilution) 6 4 2 20 0 D. rubidis F. nivalis P. minuta O. nitens 0 none D. rubidis F. nivalis P. minuta O. nitens

Species sensitivity distribution 99.9 99 Percentile of Rank Distribution Percentile of Rank Distribution 90 70 50 30 10 99.9 99 90 70 50 30 10 1 Standard and Boreal Species Standard Species Boreal Species Standard Species Boreal Species F3: 45,500 mg/kg F3: 10,600 mg/kg 1 0.1 0.1 0.1 1 10 100 % dilution 0.1 1 10 100 % dilution

Research Goal Soil Test Method Development (ecological-relevance & applicability) Collected reference and contaminated soils representative of Canadian boreal, northern and wetland regions Established cultures of candidate soil invertebrate species and seed sources Taxonomic verification traditional and genetic sequencing methods Initiated soil toxicity test method development using reference soils Discovered that some species performance is limited in some soil types; we need to define limits of non-contaminant factors so that appropriate test batteries can be developed for soil-species scenarios Ongoing development of test method using established wetland plant species Completed guidance document on collection of contaminated soils for site-specific risk assessments using biological tests

Future Goals: Complete development of ecologically-relevant tools for site assessments and evaluation of remedial techniques Boreal and taiga terrestrial plant species (published 2013); wetland plant species (2014) Soil invertebrate species in 2013 (collembolan) and in 2014 (earthworm and mite) Soil microbial tests (2014) Evaluation of effects on biomass, activity, diversity and community structure in natural consortium

Questions? Mary Moody moody@src.sk.ca