Using Bioassays to Evaluate the Performance of Risk Management Techniques
Carolyn Acheson, M. Christina Brinkman, and Gregory Sayles
Common Risk Management Assumptions
.Risk -characterized by contaminants -Ignores
	.Incomplete removal or side products
	.Co-Contaminants
	.Matrix Effects
.Treatment reduces toxicity -Ignores
	.Process Amendments
	.Other reactions
	.Matrix Changes
Case Study 1
.Remediation of PCB Contaminated Soil by Solvent Extraction
.Principals: MarkMeckes, John Meier, andLinaChang
.More information -Meier, et al. 1997.
Environmental Toxicology and Chemistry, p. 928-938.
Case Study 1 - Chemical Analysis
.PCBs
.VOCs
.SVOCs
.Metals by TCLP
Case Study 1 -Bioassays
Bioassay: Earthworm Survival Organism:E. fetidaL. terrestris Exposure Matrix: soil Exposure Period: 14 days Endpoint: Survival
Bioassay: Seed Germination Organism: Oats andLettuce Exposure Matrix: soil Exposure Period:5 days Endpoint: Survival
Bioassay: Earthworm Reproduction Organism: E. fetida Exposure Matrix: soil Exposure Period: 3 weeks Endpoint: Survival, body mass,number of cocoons,cocoon hatchability
Bioassay: Root Elongation Organism: Oats andLettuce Exposure Matrix: soil5 daysGrowth
Bioassay: Allium Mitotic Aberrations Organism: Allium Exposure Matrix: water extract Exposure Period: 24 hour Endpoint: Mitotic index,chromosomalabnormalities
Case Study 1 -Solvent Extraction
Solvent
Solvent +PCBs
Soil+ PCBs
Soil
Case Study 1 -Results
Chart by PCB Concentration (mg/kg soil) and Before Treatment After Treatment Control
Contact Carolyn Acheson at (513) 569-7190
Case Study 1 -Results
Chart by PCB Concentration (mg/kg soil) and Before Treatment After Treatment Control
Contact Carolyn Acheson at (513) 569-7190
Case Study 1 -Better RM
Solvent
Solvent
+PCBs
Soil+ PCBs
Soil
Water
Water+
2-Propanol
Case Study 1 -Results 2
Chart by PCB Concentration (mg/kg soil) and Before Treatment After Treatment Control
Contact Carolyn Acheson at (513) 569-7190
Case Study 1 -Summary
.Solvent extraction removed PCBs from soil
.Process residues were as toxic as PCBs
.Better RM -Add rinse step
	.Reduce PCB concentration
	.Reduce toxicity
Case Study 2
.Remediation of soil contaminated with wood treating wastes by Soil Washing
	.Fluid -Ethanol-water mixture
	.Question -2 or 3 Soil Washing stages?
.Principals
	.Soil Washing -Richard Brenner, Makram Suidan, George Sorial, Amid Khodadoust, Karen Koran, and Gregory Wilson
	.EcotoxicityEvaluation -Carolyn Acheson, Jennifer
Mansfield , YongguiShan, and Margaret Kupferle
Case Study 2
.Chemical Analysis
	.PCP
	.Hydrocarbons -alkanes and PAHs
.Bioassays
	.Earthworm Survival
	.Seed Germination and Root Elongation in Lettuce and Oats
Case Study 2 -Soil Washing
2 -stage
Treated
Soil
Contaminated
Soil
Rinse
1
H2O
Rinse
2
H2O
Rinse
1
H2O
Rinse
2
H2O
Wash 1
Wash 2
Wash 3
Wash 1
Wash 2
3-stageTreatedSoil
Case Study 2 -Results
Chart
Contact Carolyn Acheson at (513) 569-7190
Case Study 2 -Results
Chart
Contact Carolyn Acheson at (513) 569-7190
Case Study 2 -Results
Chart
Contact Carolyn Acheson at (513) 569-7190
Case Study 2 -Results
Chart
Contact Carolyn Acheson at (513) 569-7190
Case Study 2 -Summary
.Soil Washing was effective
	.Chemistry
		.PCP, Alkanes, and PAHs removed
		.3-stage process most effective
	.Bioassays
		.Earthworms and plants show reduced toxicity in
treated soils.
		.2-stage process most ecologically hospitable
.Likely that soil washing alters other aspects of soil
Risk Management of EDCs
.Uncertainties of EDCs
	.Unknown endocrine activity of degradation products
	.Unknown effectiveness of treatments in reducing endocrine activity
.Concurrent chemical and biological measures of effectiveness recommended
EDC Bioassays -Considerations
.EDCs of concern in NRMRL projects
.Concentrations
.Environmental Matrices
.Data Quality -Reproducibility and Reliability
.Practicality -Cost and Ease of Use
.Recommendations of Others
.Adaptability to RM projects
EDCsof Concern in NRMRL projects
.Alkylphenols
.Chlorinated Dioxins and Furans
.Estrogens, biogenic and pharmaceutical
.PCBs
All are estrogenic;some have thyroid and developmental effects
EDC Bioassays -Considerations
.Environmental Matrices in NRMRL projects
	.Air
	.Water
	.Solids -soils, sediments, and biosolids
.Concentrations
	.Water -as low as ng17 $-estradiol/L
	.solids -levels vary
EDC Bioassays - Considerations
.Data quality
.Practicality
.Adaptability
.Sensitivity
Evaluated by:
Peer reviewed literature
EDSTAC report
ORD colleague recommendations
Types of EDC Bioassays Considered
.Sediment/Aquatic Invertebrate tests
.Terrestrial Invertebrate tests
.In vitrotests
FishVitellogenin mRNAassay through
cooperation with MERB/NERL
Sediment/Aquatic Invertebrate tests
.Advantages
	.Commonly studied aquatic organisms
	.Many endpoint options
.Disadvantages
	.Mechanism of action -interference with molting controlled by steroid hormones, ecdysteroids
	.Require substantial lab equipment
	.Test duration -about 1 month
Image From www.AquacultureStore.com
Terrestrial Invertebrate tests
.Imposex Occurrence -measure of androgenicity
.Earthworm Reproduction
	.methods exist
	.endpoints such as number of cocoons and number of hatchlings per cocoon
	.endpoints are not directly related to endocrine function
	.unknown sensitivity to EDCs of concern
In VitroAssays
.Mammalian cells: E-Screen and MVLN
	.organism -immortal mammalian cell (MCF-7)
	.endpoint -proliferation or luciferaseproduction
	.MVLN -recommended by EDSTAC
.Yeast Estrogen Screening Assay (YES)
	.evaluated by EDSTAC
	.commonly used in peer reviewed literature
	.not recommended for chlorinated pesticides
.Both -reported sensitivity at low concentrations
EDC Bioassay Selected for Adaption -
YES Assay
Chart
Contact Carolyn Acheson at (513) 569-7190
NRMRL Sponsored EDC RM Projects
Using Bioassays
Project: EDC: Prinicpal Investigator: Bioassay:
Project: Evaluation of Drinking Water TreatmentTechniques for EDC Removal  EDC: Steroid Hormones Alkyl Phenols  Prinicpal Investigator:Kathleen Schenck Bioassay: MVLN Assay
Project: Potential of CAFOs to ContributeEstrogens to the Environment  EDC: Estrogens  Prinicpal Investigator:StevenHutchins Bioassay: FETAXXTRA
Project: Investigations of Sorption and Transport ofHormones and Animal Pharmaceuticals  EDC: Estrogens   Prinicpal Investigator:Carl Enfield and Suresh Rao Bioassay: YES Assay
Project: Evaluating the Fate of EDCs During Wastewater Treatment Steroid Hormones Alkyl Phenols  Prinicpal Investigator:PaulMcCauley Bioassay: YES AssayVitellogeninm RNA Assay
Project: EDCs from Combustion and VehicularEmissions PCBsDioxins/Furans  Prinicpal Investigator:Brian Gullett Bioassay: VitellogeninmRNA Assay
Project: Natural Recovery of PCBs in Sediments PCBs  Prinicpal Investigator: Richard Brenner Bioassay: VitellogeninmRNA Assay
Using Bioassays in a Hypothetical EDC RM Project
.EDC of Concern -Phthalates
	.commonly used as a plasticizer in many household products (including food containers)
	.suspected to cause alterations in human sexualdevelopment
.RM Project -Find a replacement plasticizerfor phthalates
Hypothetical EDC RM Project -Phthalate Replacement
.Use computer models to find substances with appropriate chemical and physical properties
.Lab Testing of leading candidates
	.chemical and physical testing to determine if acceptable substitute
	.bioassays to evaluate biological activity
.Look at production processes
	.Are production by-products likely to cause problems?
	.Test bulk chemical -chemical, physical, biological properties
Acknowledgements
.Jennifer Mansfield, IT Corporation
.Kathleen Schenck, U.S. EPA
.Andrew Avel, U.S. EPA