1. Tyler Mehler1, Jing You2, Jon Maul3 and Michael Lydy1
Identification of the Causes of Sediment-Associated Toxicity in the Illinois River Complex Using a Sediment-TIE Approach
Tyler Mehler1, Jing You2, Jon Maul3 and Michael Lydy1
1Fisheries and Illinois Aquaculture and Department of Zoology, Southern Illinois University, Carbondale, Illinois, USA
2State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou, China
3The Institute of Environmental and Human Health, Department of Toxicology, Box 41163, Texas Tech University, Lubbock, Texas, USA
ISTC seminar: 9/9/09

2. Amended for non-polar organics
What is a TIE?
As defined by the EPA (2007):
The Toxicity Identification Evaluation approach “is to use physical/chemical manipulations of a sample to isolate or change the potency of different groups of toxicants potentially present in a sample”.
Contaminated w/ non-polar organics X X X X X X X X X
Amended for non-polar organics
Amended for ammonia
Unamended
Amended for metals
Amended

3. Matrix Choice Sediment grain Pore water Issues with pore water TIEs:
usgs.gov
Issues with pore water TIEs:
Bioavailability
Ingestion
Water quality parameters
Environmental Realistic?

4. Conducting a Toxicity Identification Evaluation (TIE)
STEP 1
SITE SAMPLING
STEP 2
SCREENING TOXICITY TEST
STEP 3
PHASE I: CHARACTERIZATION
Ammonia (Zeolite)
Cationic Metals (Resin-Tech SIR 300) X
Nonpolar Organics (PCC)
Unamended
Amended
STEP 4
PHASE II: IDENTIFICATION
Ammonia (Ammonia Probe)
Cationic metals (NAD & FAAS)
Nonpolar Organics (ASE & GC/HPLC)
concentration of contaminant
LC50 of that contaminant
Toxic Unit (TU) =
U.S. EPA 2007 Whole sediment TIE guidelines

5. Objectives Identify toxic sites throughout the Illinois River Complex
Identify the contaminant classes (ammonia, metals, non-polar organics) that attribute to the toxicity of those sites using a whole-sediment TIE test
Evaluate the temporal and spatial trends in correlation to the toxicity of those sites
Examine the difference between TIE methodologies and the test organisms used
Compare past and present TIE research on the IRC
The objective of this study is quite similar and also attempts to address a few new components. READ.

6. Conducting a Toxicity Identification Evaluation (TIE)
STEP 1
SITE SAMPLING
SITE SAMPLING
STEP 2
SCREENING TOXICITY TEST
STEP 3
PHASE I: CHARACTERIZATION
Ammonia (Zeolite)
Cationic Metals (Resin-Tech SIR 300)
Nonpolar Organics (PCC)
STEP 4
PHASE II: IDENTIFICATION
Ammonia (Ammonia Probe)
Cationic metals (NAD & FAAS)
Nonpolar Organics (ASE & GC/HPLC)

7. Sampling Methods 24 sites chosen with consultation of ISTC17
Sampling Methods
24 sites chosen with consultation of ISTC
2.5 kg was collected from each site
Water samples from each site were also retrieved and water quality measurements for each site taken
Hardness emulated
Total pore water ammonia was analyzed upon arrival at SIUC
Sediments and water samples were analyzed at SIUC Fisheries and Illinois Aquaculture Center
Samples were taken in summer 07’, fall 07’, winter 07-08’, spring 08’ and again in summer 08’

8. Sites Rivermile 17
St. Louis
Carbondale X

9. Conducting a Toxicity Identification Evaluation (TIE)
STEP 1
SITE SAMPLING
STEP 2
SCREENING TOXICITY TEST
SCREENING TOXICITY TEST
STEP 3
PHASE I: CHARACTERIZATION
Ammonia (Zeolite)
Cationic Metals (Resin-Tech SIR 300)
Nonpolar Organics (PCC)
STEP 4
PHASE II: IDENTIFICATION
Ammonia (Ammonia Probe)
Cationic metals (NAD & FAAS)
Nonpolar Organics (ASE & GC/HPLC)

10. Screening Toxicity Tests
10-d bioassays (U.S. EPA) in flow-thru system with three water changes per day (100 ml per change)
10 H. azteca (14 to 21-d old) per 300 ml beaker, 6 replicates per site
Control: Touch of Nature (TON) hydrated soil – Carbondale, IL
Amendment Reference: Lower Peoria Lake (LPL)
Statistical Analysis: Dunnett’s Multiple Comparison Test

11. Summer 07’ Results: Screening Toxicity Tests
Halstead
Touch of Nature
Moore’s Towhead
Spring Lake Wildlife Area
Pekin
Wesley
Upper Lower Peoria Lake
LPL Dredge
Goose
Lacon
Mudd
Hennepin Down River
Hennenpin Power Plant
DuPue
Marseilles
DuPage
CS305
SS308
Stony Creek
SS315
SS317
SRCALRR
Sites that were significantly different from controls (p<0.05) and were chosen for seasonal analysis.

12. Conducting a Toxicity Identification Evaluation (TIE)
STEP 1
SITE SAMPLING
STEP 2
SCREENING TOXICITY TEST
STEP 3
PHASE I: CHARACTERIZATION
PHASE I: CHARACTERIZATION
Ammonia (Zeolite)
Cationic Metals (Resin-Tech SIR 300)
Nonpolar Organics (PCC)
STEP 4
PHASE II: IDENTIFICATION
Ammonia (Ammonia Probe)
Cationic metals (NAD & FAAS)
Nonpolar Organics (ASE & GC/HPLC)

13. Phase I: Characterization
20% (12 g)
Zeolite
20% (12 g)
Unamended (sand)
4 -d static test
10 H. azteca
6 reps per treatment
25% (15 g)
10-d flow-thru test
RT SIR 300 HP
15% (9 g)
PCC
25% (15 g)
Unamended (sand)

14. Summer 2007 Results: Phase I
Unamended (sand)
Organics (PCC)
TON
LPL
SS308
Halstead
CS305
Stny Crk
SRCALRR
SS315
DuPage
TOXIC SITES
The addition of zeolite (ammonia) and Resin-Tech SIR 300 (metals) showed no significant differences in comparison to the unamended sediment.

15. Conducting a Toxicity Identification Evaluation (TIE)
STEP 1
SITE SAMPLING
STEP 2
SCREENING TOXICITY TEST
STEP 3
PHASE I: CHARACTERIZATION
Ammonia (Zeolite)
Cationic Metals (Resin-Tech SIR 300)
Nonpolar Organics (PCC)
STEP 4
PHASE II: IDENTIFICATION
PHASE II: IDENTIFICATION
Ammonia (Ammonia Probe)
Cationic metals (NAD & FAAS)
Nonpolar Organics (ASE & GC/HPLC)

16. Nonpolar Organic Toxicants
Phase II: Identification
Nonpolar Organic Toxicants
PAHs: acenaphthene, acenapthylene, anthracene, chrysene, fluoranthene, fluorene, naphthalene, phenanthrene, pyrene, benzo[a]anthracene, benzo[b]fluoranthrene, benzo[k]fluoranthene, benzo[a]pyrene, benzo[g,h,i]perylene, dibenzo[a,h]anthracene and indeno[1,2,3-cd]pyrene
PCBs: Congeners: 8, 18, 28, 31, 43, 44, 48, 49, 52, 66, 70, 86, 87, 95, 97, 99, 101, 105, 110, 114, 118, 123, 126, 128, 138, 153, 156, 157, 167, 169, 170, 174, 180, 183, 187, 189, 194, 195, 200, 201, 203 and 206.
OCPs: alpha-BHC, beta-BHC, gamma-BHC, delta-BHC, p,p’-DDE, p,p’-DDD, p,p’-DDT, aldrin, gamma-chlordane, alpha-chlordane, diedrin, endrin, endrin aldehyde, endrin ketone, endosulfan I, endosulfan II, endosulfan sulfate, heptachlor, heptachlor epoxide and methoxychlor.
OP/Pyrethroids: chlorpyrifos, permethrin, lambda-cyhalothrin, cypermethrin, esfenvalerate, deltamethrin, cyfluthrin, bifenthrin.
Heavy Metals
Cu2+, Ni2+, Cr6+, Pb2+, Zn2+, Cd2+
Ammonia
NH4+, NH3

17. The reasons we analyzed total ammonia:
It’s commonly performed in standard TIE methods
Allows comparisons in pore water ammonia concentrations between past and present studies (Sparks and Ross 1992 – Concentrations measured as total ammonia)
Allows comparisons in pore water ammonia concentrations among sites (since water quality characteristics differed among sites)
Difficult to account for drifting pHs or changing temperature throughout studies

18. Summer 2007 Phase II: Identification
LPL
SS308
Halstead
CS305
Stony
Creek
SRCALRR
SS315
åMetals (µg/g dry)
6.39
37.7
26.5
27.6
50.4
23.8
17.9
Pore water total
ammonia (mg N/L)
6.48
36.6
26.2
13.7
19.4
21.7
541
åPesticides (µg/g OC)
BRL
0.447
0.208
0.414
2.14
1.12
0.405
åPCBs (µg/g OC)
4.45
11.5
15.7
21.1
37.2
34.5
7.6
åPAHs (µg/g OC)
586.8
1934
1328
1198
1021
1267
4112
Toxic Units
LPL
SS308
Halstead
CS305
Stony
Creek
SRCALRR
SS315
∑Metals (µg/g dry)
<0.1
Pore water total ammonia (mg N/L)
0.4
PCBs (µg/g oc)
Pesticides
(µg/g oc)
PAHs (µg/g oc)
0.7
2.8
1.9
1.8
1.4
1.6
4.6
BRL – 1 µg/kg
Low TU = Low Toxicity
High TU= High Toxicity

19. Summer 2007 Conclusions
Phase I findings strongly suggests that non-polar organics are the problem, with Phase II findings further suggesting that PAHs were at high concentrations to cause the noted toxicity.
What about the other seasons?

20. Is PCC always effective?
SUMMER 07’
SUMMER 08’
Affect on PCC binding capability?
Organics higher affinity for UCM?
Causes toxicity itself?
% Unresolved Complex Matrix (UCM)
Unamended (sand)
46% of sites (in all seasons) were characterized with PCC
Organics (PCC)
Is PCC always effective?

21. Phase II: Spatial and temporal variation
Total Pore Water Ammonia
Summer 2007
Fall 2007
Winter
Spring 2008
Summer 2008
mg N/L
Calumet Sag Channel
Chicago Sanitary and Shipping Canal
Total Cationic Metals
Concentration of Contaminant
µg/g dry wt
Total PAHs
6.0
mg/g OC
3.0
SS315
SS308
CS305
SRCALRR
Stony Creek
LPL
Halstead

22. Spatial Trends: Ammonia

23. Total Ammonia >400 mg N/L
Spatial Trends: Ammonia
Municipal Waste Plant
SS315
Total Ammonia >400 mg N/L
Courtesy of

24. Sparks and Ross (1992)
Gradient of increased toxicity associated with the total ammonia concentration
Ammonia the primary source of toxicity in the Illinois River Complex
Patches of toxicity occurring due to PAHs

25. VS VS A Comparison Study:
Determine differences between pore water TIE testing and whole-sediment TIE testing
Determine differences between test organisms (H. azteca and C. dubia)
While still comparing past and present research
Two sites being evaluated:
SS315 – highest ammonia concentrations
SS308 – highest PAH concentrations VS VS

26. Phase I: Pore Water Characterization
Diluted by 50%
Unamended
2-d static test
1-d static test
Zeolite
5 H. azteca
10 C. dubia
8 reps per treatment
SPE C18

27. Phase I: Whole Sediment Characterization
Zeolite
4-d static test
Unamended
PCC
10-d flow-thru test
10 H. azteca
10 C. dubia
8 reps per treatment
Zeolite
Unamended
2-d static test
PCC

28. Comparing Methodologies: Ammonia
Whole Sediment TIE
Pore Water TIE
PHASE I:
2.56
0.27
Predicted TUs for H. azteca
PHASE II:
SS315 Ammonia
Concentrations (mg N/L)
359 mg N/L
37.9 mg N/L

29. Comparing Methodologies: Non-polar Organics
PHASE I:
Whole-Sediment TIE
Pore Water TIE
The affects of:
UCM
Black carbon
ingestion, adsorption
DOC
Glassware binding
0.52
1.81
TUs for H. azteca
PHASE II:
SS308 ∑PAH Concentrations:
1953 µg/L
4405 µg/g oc

30. Comparing Species Sensitivity/Susceptibility
H. azteca
 140 mg N/L (4-d)a
LC50 Total Ammonia:
C. dubia
 mg N/L (2-d)b
H. azteca
 30.6 µg/L (10-d)c
LC50 Fluoranthene (PAH):
C. dubia
 µg/L (10-d)c
a – Ankley et al. 1995
b – extrapolated from Bailey et al. 2001
c – Suedel and Rodgers, Jr. 1996
Species Sensitivity ≠ Species Susceptibility
Body Size /Age (Life Stage)
Physiology/Feeding Behavior
Niche

31. Conclusions
Toxic sites were identified on the IRC for future risk assessment & mitigation
Rm 277 (DuPage) Calumet Sag Channel, Chicago Sanitary and Shipping Canal

32. Conclusions
PAHs and the associated oils and grease were identified as the sources of the noted toxicity, however ammonia was elevated at SS315

33. Conclusions
Little temporal variation was noted in toxicity and in concentrations
However, spatial trends were found in toxicity especially concerning ammonia

34. Conclusions Which TIE approach is better and where are TIEs headed?
Is the IRC a healthy system ?

35. Acknowledgements Sampling Crew:
Ed Workman, Mandy Rothert, Liz Tripp, Heather Foslund
A special thanks goes to SETAC (Student Exchange Program) and Teresa Norberg-King and the rest of the EPA Duluth Lab.
Fisheries and Aquaculture Center and Dept. of Zoology staff and students
Funding:

36. For more information:
Mehler WT, Maul JD, You J, and MJ Lydy Identifying the causes of sediment-associated contamination in the Illinois River using a whole-sediment Toxicity Identification Evaluation (TIE). Environmental Toxicology and Chemistry. In Press. Mehler WT, You J, Maul JD, and MJ Lydy Comparative analysis of whole sediment and pore water Toxicity Identification Evaluation (TIE) techniques for ammonia and non-polar organic contaminants. Chemosphere. In Review.

40. Questions?

41. Evaluating Ammonia SS315 overlying water SS315 pore water
Total ammonia – 37.9 mg N/L (TU = 0.29)
Un-ionized ammonia – mg/L (TU = 0.27)
SS315 pore water
Total ammonia – 359 mg N/L (TU = 2.56)
Un-ionized ammonia – 11.2 mg/L (TU = 5.19)

42. Sparks and Ross (1992)
Gradient of increased toxicity associated with the total ammonia concentration
Ammonia was the primary source of toxicity in the Illinois River Complex
Patches of toxicity occurring due to PAHs
The beginning of a general recovery of the Illinois River Complex
One study by Sparks and ross attempted to address the sources of toxicity in the IRC and the risk that this contamination may pose using a pore water tie. I will discuss what a tie is shortly, but these were the major findings of there study approx. 20 years ago. READ.

44. Sparks and Ross 1992
Gradient of increased toxicity associated with the total ammonia concentration
Ammonia the primary source of toxicity in the Illinois River Complex
Patches of toxicity occurring due to PAHs
The beginning of a general recovery of the Illinois River Complex ?

45. Questions?

46. Conclusions:
Differences in past and present TIE studies is attributed to the differences in methodologies and perhaps on a lesser note test organism choice.
Which test organism is better?
Realistic Test Organism?
Using Historic Test Organism?
Which TIE is better and where are TIEs headed?