muskrats. Also, roots and leaves <strong>of</strong> aquatic plants can be consumed bybenthic omnivorous fish, such as common carp, catfish, white perch, etc.However, some bioaccumulation studies can be performed with much smallerorganisms (e.g., springtails or potworms), depending on study objectives.Individual springtails or potworms are very small, requiring hundreds <strong>of</strong>organisms to make up a 50-gram tissue mass requirement. Because it isusually not feasible to set up multiple replicate samples with hundreds <strong>of</strong>organisms, it is best to limit use <strong>of</strong> smaller invertebrates to soil samples withCOPEC lists with a small analytical tissue requirement.Laboratory QA/QC ObjectivesReplicatesAs is required with any toxicity study, appropriate replication is necessary toensure comparability between soil or sediment samples and laboratory controlor reference samples. Bioaccumulation studies should be designed to producethe required analytical tissue mass from each test replicate. If sufficient tissuemass is not possible from each replicate, compositing between replicates isacceptable. However, compositing reduces the statistical power <strong>of</strong> the study.If all test replicates have to be composited to a single sample, statisticalcomparisons are not appropriate and the study may not meet the data qualityobjectives for inclusion in a risk assessment. Where sufficient historical dataare available, a power analysis (Zar, 1984) is <strong>of</strong>ten useful to determine howmany test replicates may be necessary to ascertain differences between soil orsediment sampling locations or reference area soils.Tissue Preparation and ProcessingBecause even a small amount <strong>of</strong> potentially contaminated soil or sediment cansignificantly bias any tissue analysis, it is critical to remove all traces <strong>of</strong> soilor sediment from the test organisms before submitting the tissue for COPECanalysis. For example, plant roots should be thoroughly rinsed and carefullyexamined to ensure that all soil is removed. Earthworms and other organismsthat ingest and process soil should be depurated for a time sufficient to ensurethat the digestive tract is empty. While 24 hours is an acceptable depurationperiod (ASTM, 2004), studies have indicated that different species mayrequire 48 hours for sufficient depuration, and some other species may requiredissection and rinsing <strong>of</strong> the digestive tract (Arnold and Hodson, 2007).All tissue samples should be analyzed for COPECs, percent moisture andpercent lipids (as appropriate). In addition to the analysis <strong>of</strong> tissue samples,bioaccumulation studies should include analysis <strong>of</strong> all site soil and/orsediment samples, reference samples and laboratory control samples forCOPECs, grain size, TOC and pH. Soils used for plant toxicity orbioaccumulation studies should also be analyzed for soil nutrientconcentrations.<strong>Ecological</strong> <strong>Evaluation</strong> <strong>Technical</strong> <strong>Guidance</strong> Document 45Version 1.2 8/29/12
While there are many laboratories that can produce acceptable analyticalresults for soil and sediment samples, not all laboratories are capable <strong>of</strong> tissueanalysis. Tissue processing requires specialized equipment in addition to thestandard analytical instrumentation, and some laboratories cannot work withsmall samples. Tissue samples, particularly samples with high lipid content,may also present analytical interference that can yield excessively highanalytical detection limits for insufficiently experienced laboratories.It is critical to discuss bioaccumulation study objectives with both the toxicitytesting laboratory and the analytical laboratory, to ensure that data qualityobjectives are met.6.1.3.3 Toxicity Reference ValuesToxicity reference values (TRV) are literature-based levels defined as a doseabove which ecologically relevant effects might occur in wildlife speciesfollowing chronic dietary exposure and below which it is reasonably expectedthat such effects will not occur (USEPA, 2005a). TRVs provide a basis forestimating whether the exposure to COPECs at a site is likely to result inadverse ecological effects (e.g., survival, growth, and reproduction <strong>of</strong> wildlifespecies). The TRVs selected in the ERA are based on LOAELs and/orNOAELs from laboratory and/or field-based studies reported in the scientificliterature (USEPA, 2005a and Sample et al., 1996).Risks are estimated for appropriate feeding guilds by comparing the results <strong>of</strong>the measured or modeled dietary contaminant doses to receptors <strong>of</strong> concern tothe TRVs. The ratio <strong>of</strong> these two numbers is called a Hazard Quotient (HQ).HQs equal to or greater than one (HQ ≥ 1) are typically considered to indicatepotential risk to ecological receptors. If an HQ suggests that effects are notexpected to occur for the average individual, then the effects are likely to beinsignificant at the population level; however, if an HQ indicates risks arepresent for the average individual, then risks may be present for the localpopulation.The basic equation used for calculation <strong>of</strong> an HQ value for exposure <strong>of</strong> awildlife receptor to a chemical by ingestion <strong>of</strong> an environmental medium is:HQi, j, r = ADDoTRVi,rwhere:HQi,j,r = HQ for exposure <strong>of</strong> receptor “r” to COPEC “i” in medium “j”ADD = Average Daily Dose (mg/kgBW-day)oTRVi,r = Oral TRV for COPEC “i” in receptor “r” (mg/kg-day)Because all receptors are exposed to more than one environmental medium,the total hazard to a receptor from a specific COPEC is calculated as the sum<strong>of</strong> HQs for that COPEC across all media:HQi, r = Σ HQi, j, r<strong>Ecological</strong> <strong>Evaluation</strong> <strong>Technical</strong> <strong>Guidance</strong> Document 46Version 1.2 8/29/12
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http://www.epa.gov/owow/oceans/regu
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USEPA 2006a. Data Quality Assessmen
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Appendix A - Habitat Survey FormsEc
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Ecological Evaluation Technical Gui
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Appendix B - Sampling Procedures fo
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Appendix C - Surface Water Toxicity
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Short-term chronic studies, endpoin
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Appendix D - Sediment Toxicity Test
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Toxicity Test DesignSediment toxici
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Appendix E - Sediment Pore Water an
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The seven-day daphnid survival and
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esults are then evaluated using USE
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Surber or Square-foot BottomThis sa
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Appendix H - Soil Toxicity TestingS
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another sample may still have a sub
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conservative approach from an ecolo
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Data PresentationTabular presentati