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T VA - K I N G S TO N F LY A S H R E L E A S E E N V I R O N M E N TA L R E S E A R C H S Y M P O S I U M
M A R C H 11 – 1 2 , 2 0 1 0
R I C K M . S H E R R A R D , P h . D .
T E N N E S S E E VA L L E Y A U T H O R I T Y
Ecotoxicology Overview
TOPICS
EcotoxicologyStudies conducted by TVA independent laboratoriesBrief summary results of studies to dateStudies conducted by other laboratoriesOngoing and future studies plannedLaboratory challenges in working with ashGaps in knowledge
What is Ecotoxicology?
“In the broadest sense of the word, an ecotoxicologist is one who carries out toxicity testing on any component of any ecosystem.” Cairns, 1989
Ecotoxicity studies are designed to employ ecological attributes to assess toxicity
Goal of the assessment – to protect entire ecosystems, not isolated components
TVA-Sponsored StudiesConducted by Independent Laboratories
Phase I: April through June 2009 Vibracore® Dredge Plume Plant Stilling Pond Effluent (Outfall 001)
Phase II: August 2009 to present Dredge Plume Plant Stilling Pond Effluent (Outfall 001)
Focus: Assess the potential for short-term effects associated with
removing ash from the Emory River
Phase ISampling and
Analyses
IV. Toxicological Monitoring
A. Whole Sediment Toxicity Evaluation
B. Elutriate Toxicity Evaluation
C. Plume Toxicity Evaluation
D. Polymer Toxicity Evaluation (Stilling Pond Effluent)
Samples
Vibracore® upstream and downstream
Vb.1, Vb.2 – 3/17/09
Vb.3, Vb.4 – 6/11-12/09
Cores composited, homogenized
Stored in 19-L plastic buckets at 4ºC
Holding time = 8 weeks
Re-homogenized prior to use
Lab control CRM 189.0
Unaffected Emory River water controls, overlay, diluent
Hyalella azteca Survival and Growth Test for Sediments
Duration: 10 d
Temperature: 23 ± 1ºC
Photoperiod: 16 / 8
Sed. Volume: 100 mL
Water Volume: 175 mL
Renewal: 2 vol. add./d
Age: 7 – 14-d
No./Chamber: 10
No. Replicates: 8
Feeding: YCT 1 mL/d
Endpoints: Survival, Growth
Acceptance: ≥ 80%
Freshwater Juvenile MusselSurvival Test for Sediments
(Lampsilis siliquoidea, L. cardium)
Duration: 5 d & 10 d
Temperature: 24 ± 1ºC
Photoperiod: 24 dark
Sed. Volume: 20 mL
Water Volume: 200 mL
Renewal: 5d-0 10d-1
Age: < 8 d
No./Chamber: 10
No. Replicates: 5
Feeding: every 3 d
Endpoint: Survival
Acceptance: ≥ 90%
Lumbriculus variegatus Survival Test for Sediments
(qualifying test for 28-d bioaccumulation test)
Duration: 4 d
Temperature: 23 ± 1ºC
Photoperiod: 16 / 8
Sed. Volume: 100 mL
Water Volume: 175 mL
Renewal: 2 vol. add./d
Age: Adults
No./Chamber: 10
No. Replicates: 4
Feeding: None
Endpoint: Burrowing, Survival
Acceptance: ≥ 90%
Corbicula fluminea Bioaccumulation Test for Sediments
Duration: 28 d
Temperature: 20 ± 1ºC
Photoperiod: 16 / 8
Sed. Volume: > 5 L
Water Volume: 15 L
Renewal: 50% 3/wk
Size: 0.5-1.5 g WTW
No./Chamber: ~ 30
No. Replicates: 5
Feeding: None
Endpoint: Bioaccum.
Acceptance: Mass
Ceriodaphnia dubia Survival Testfor Elutriates
Duration: 96 h
Temperature: 25 ± 1ºC
Photoperiod: 16 / 8
Water Vol.: 15 mL min
Renewal: None
Age: < 24 h
No./Chamber: 5
No. Replicates: 5
Feeding: Prior / 48 h
Concentrations: Non-Centrifuged 5 (0-100%); Centrifuged 1 (100%)
Controls: ERW, MHSW
Endpoint: Survival
Acceptance: ≥ 90%
Pimephales promelas Survival Testfor Elutriates
Duration: 96 h
Temperature: 25 ± 1ºC
Photoperiod: 16 / 8
Water Vol.: 200 mL min
Renewal: None
Age: < 24 h
No./Chamber: 10
No. Replicates: 5
Feeding: Prior / 48 h
Concentrations: Non-Centrifuged 5 (0-100%); Centrifuged 1 (100%)
Controls: ERW, MHSW
Endpoint: Survival
Acceptance: ≥ 90%
Freshwater Juvenile MusselSurvival Test for Elutriates
(Lampsilis siliquoidea, L. cardium)
Duration: 10 d
Temperature: 24 ± 1ºC
Photoperiod: 24 dark
Sed. Volume: 20 mL clean
Water Volume: 200 mL
Renewal: 1 (day 6)
Age: < 8 d
No./Chamber: 10
No. Replicates: 5
Feeding: every 3 d
Concentrations: Non-Centrifuged 5 (0-100%); Centrifuged 1 (100%)
Controls: ERW, MHSW
Endpoint: Survival
Acceptance: ≥ 90%
Ceriodaphnia dubiaSurvival and Reproduction Test
for Emory River Plume and Outfall 001
Duration: 3 broods
Temperature: 25 ± 1ºC
Photoperiod: 16 / 8
Water Vol.: 15 mL min
Renewal: Daily
Age: < 24 h
No./Chamber: 1
No. Replicates: 10
Feeding: Once daily
Concentrations: 5 (0-100%)
Controls: ERW, MHSW
Endpoints: Survival, Reproduction
Acceptance: ≥ 80% survival, ≥15 neos/survivor
Pimephales promelas Survival and Growth Test for Emory River Plume
and Outfall 001
Duration: 7 d
Temperature: 25 ± 1ºC
Photoperiod: 16 / 8
Water Vol.: 250 mL min
Renewal: Daily
Age: < 24 h
No./Chamber: 10
No. Replicates: 4
Feeding: Twice daily
Concentrations: 5 (0-100%)
Controls: ERW, MHSW
Endpoints: Survival, Growth
Acceptance: ≥ 80% survival, 0.25 mg growth
Phase IISampling and
Analyses
Weekly (Aug – Sept)
Biweekly (Oct – present)
Plume: grab samples
Outfall 001: 24-h composites
Unaffected Emory River water controls, diluent
Ceriodaphnia dubia Survival Testfor Emory River Plume and Outfall 001
Duration: 96 h
Temperature: 25 ± 1ºC
Photoperiod: 16 / 8
Water Vol.: 15 mL min
Renewal: At 48 h
Age: < 24 h
No./Chamber: 5
No. Replicates: 5
Feeding: Prior / 48 h
Concentrations: 5 (0-100%Controls: ERW, MHSW
Endpoint: Survival
Acceptance: ≥ 90%
Pimephales promelas Survival Testfor Emory River Plume and Outfall 001
Duration: 96 h
Temperature: 25 ± 1ºC
Photoperiod: 16 / 8
Water Vol.: 200 mL min
Renewal: At 48 h
Age: < 24 h
No./Chamber: 10
No. Replicates: 5
Feeding: Prior / 48 h
Concentrations: 5 (0-100%)
Controls: ERW, MHSW
Endpoint: Survival
Acceptance: ≥ 90%
Vb.1, Vb.2 Ash Composites Vb.3, Vb.4 Ash Composites
Whole ash H. azteca: adverse effects L. variegatus: can’t burrow L. siliquoidea: effects
limited to 1 of 4 tests C. fluminea: no
bioaccumulation
Elutriates: no effects
Whole ash H. azteca: adverse effects L. variegatus: can’t burrow L. cardium: effects in one
5-d test and one 10-d test C. fluminea: no
bioaccumulation
Elutriates: mixed bag
TVA Phase I Summary Results: Ash Samples
TVA Phase I & II Summary Results:Plume and Outfall 001 Samples
No adverse effects in tests with exposures to plume samples
Only one Outfall 001 sample to date resulted in decreased survival to C. dubia (but not P. promelas)
Pathogen interference is being dealt with appropriately through parallel testing of UV-treated and untreated test solutions
Other Ecotoxicological Studies
USACE – ERDC 10-d larval fish elutriate bioassay 10-d juvenile fish elutriate bioassay Bioaccumulation & health indicators
USGS, USFWS Whole sediment and elutriate studies with benthic
invertebrates H. azteca, Villosa iris, Lampsilis fasciola, Chironomus dilutus 10-d, 28-d
Studies Planned by TVA
Independent laboratory bioavai labi l i ty study wi th H. azteca and C. dubia
Whole ash, porewater only With and without resin treatment
ORNL ear ly l i fe stages effects study wi th f ish Task 1: Fish embryo-larval toxicity tests of fly ash Task 2: Longer-term exposures to fly ash in the laboratory Task 3: Evaluating early life stage success in fly ash exposed
fish populations
Challenges: Ecotoxicological Studies of Kingston Fly Ash Site
Site locat ionWater characterist ics of converging r iversNative sediment characterist ics Behavior of ash in storage and test chambersPathogen interference in Emory River water
Reference Control Sediment
Currently using clean sediment from Clinch River Mile 189.0 for comparisons
To date, TVA has: Attempted to formulate sediment Considered ash washing Incorporated ion-exchange resin treatment
Homogenizing Whole Ash Samples
In storage, ash settles and compacts, porewater surfaces
Water poured off
Stainless steel beater with teeth on bottom chips away at ash
Water content decreases toward bottom of container
Entire contents blended into pourable slurry
Pathogen Interference in P. promelas Exposures with Emory River Water
Well-documented in Kingston Fossil Plant NPDES WET monitoring history
Interrupted concentration-response
High variability in mortality between replicates
Greater effect on Emory River controls and lower concentrations
Ultraviolet Treatment of Test Solutions Prepared with Emory River Water
All P. promelas tests with Emory River water
Parallel exposures (with and without UV treatment)
Dilutions prepared first, split, then treated individually
Treatment durations are 2, 3.5, or 5 min based on turbidity
Interrupted concentration-response and high variation between replicates - invalidated
Pathogen Interference in L. cardium Exposures with Vb.3 and Vb. 4
Laboratory Reported:
Organism stress as early as 48 h, including MHSW controls
At 96 h, large masses of “debris” (fungal filaments with colonies of protozoa)
Test organisms entangled in debris
Sporadic mortality among replicates in most test treatments
Source could be test organism supplier or Emory River water
Suggestions
Avoid “ tunnel v is ion” – o ther const i tuents can cause effec ts
Bet ter unders tanding o f fa te character is t ics o f ash const i tuents
Ash is not a natura l sed iment but models that we use to pred ic t tox ic i ty and assess r isks are based on s tud ies wi th natura l sed iments
Remember to use a weight o f ev idence approach Conduct s tud ies in a sc ient i f i ca l ly -defens ib le manner