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