Text

Submittal of Discharge Monitoring Report for November 2011
	ML11348A101
Person / Time
Site:	Sequoyah
Issue date:	12/12/2011
From:	John Carlin; Tennessee Valley Authority
To:	Waits D; Office of Nuclear Reactor Regulation, State of TN, Dept of Environment & Conservation, Div of Water Pollution Control
References
	Download: ML11348A101 (110)
	v • d • e

Tennessee Valley Authority, Post Office Box 2000. Soddy Daisy. Tennessee 37384-2000 December 12, 2011 Ms. Dana Waits State of Tennessee Department of Environment and Conservation Division of Water Pollution Control Enforcement & Compliance Section 6{h Floor, L & C Annex 401 Church Street Nashville, Tennessee 37243-1534

Dear Ms. Waits:

SEQUOYAH NUCLEAR PLANT - DISCHARGE MONITORING REPORT FOR NOVEMBER 2011 Enclosed is the November 2011 Discharge Monitoring Report for Sequoyah Nuclear Plant. If you have any questions or need additional information, please contact Brad Love at (423) 843-6714 of Sequoyah's Environmental staff.

/ certify under penalty of law that this document and all attachments were preparedunder my direction or supervision in accordance with a system designed to assure that qualified personnel properly gather and evaluate the information submitted. Based on my inquiry of the person or persons who manage the system, or those persons directly responsible for gatheringthe information, the information submitted is, to the best of my knowledge and belief, true, accurate, and complete. I am aware that there are significant penalties for submitting false information, including the possibility of fine and imprisonment for knowing violations.

Sincerely, c re sident SoIoyah Nuclear Plant Enclosures cc (Enclosures):

Chattanooga Environmental Field Office U.S. Nuclear Regulatory Commission Division of Water Pollution Control Attn: Document Control Desk State Office Building, Suite 550 Washington, DC 20555 540 McCallie Avenue Chattanooga, Tennessee 37402-2013

PERMITIEE NAME/ADDRESS (Include Faciliht Name/Location if Different) NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

MAJOR Form Approved.

Name TVA - SEQUOYAH NUCLEAR PLANT DISCHARGE MONITORING REPORT (DMR)

(SUBR 01) OMB No. 2040-0004 Address P.O. BOX 2000

.. ..L.N1TEROFFICE OPS-5N-SQN TN0026450 ý 101 G F - FINAL SODDY - DAISY TIN 37384 PERMIT NUMBER IDISCHARGE NU ER DIFFUSER DISCHARGE Facility TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON CQUNTY MONITORING PERIOD EFFLUENT YEAR MO I DAY I YEAR I MO DAY ATTN: Brad Love From 11 11 011 To 1 11 3o NO DISCHARGE [j j NOTE: Read instructions before completinq this form.

PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO. FREQUENCY SAMPLE EX OF TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS TEMPERATURE, WATER DEG. SAMPLE

31.7 04 0 30 / 30 RCORDR CENTIGRADE MEASUREMENT 00010 1 0 PERMIT Req. Mon. DEG. C. CONTI CALCTD EFFLUENT GROSS REQUIREMENT DAILY MAX NUOUS TEMPERATURE, WATER DEG. SAMPLE ** 18.2 04 0 30/30 MODELD CENTIGRADE MEASUREMENT 00010 Z 0 PERMIT 30.5 DEG. C. CONTI CALCTD INSTREAM MONITORING REQUIREMENT DAILY MX NUOUS TEMP. DIFF. BETWEEN SAMP. & SAMPLE

2 04 0 30/30 CALCTD UPSTRM DEG.C MEASUREMENT 00016 1 1 PERMIT 5 DEG. C. CONTI CALCTD EFFLUENT GROSS REQUIREMENT DAILY MX NUOUS FLOW, IN CONDUIT OR THRU SAMPLE 1692 03 0 30/30 RCORDR TREATMENT PLANT MEASUREMENT 50050 1 0 EFFLENT ROSSREQUIREMENT PERMIT Req. Mon. MGD CONTI RCORDR EFFLUENT GROSS DAILY MAX NUOUS CHLORINE, TOTAL RESIDUAL SAMPLE 0.020 0.029 19 0 29/30 GRAB MEASUREMENT 50060 1 0 PERMIT ******** 0.1 0.1 MG/L FIVE PER CALCTD EFFLUENT GROSS REQUIREMENT MO AVG DAILY MAX WEEK TEMPERATURE - C, RATE OF SAMPLE 0 62 ** 0 30 /30 CALCTD CHANGE MEASUREMENT 82234 1 0 PERMIT 2 DEG CONTI CALCTD EFFLUENT GROSS REQUIREMENT DAILY MX ClHR NUOUS SAMPLE MEASUREMENT PERMIT REQUIREMENT NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my direction or supervision in accordance with a system designed to assure that qualified personnel I TELEPHONE DATE John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the person or persons who manage the system, or those persons directly responsible for gathering the . ( 4e pahSite Vice President information, the information submitted is, to the best of my knowledge and belief, true, accurate, 423 843-7001 11 12 09 Sequoyah Site Vice President and complete. I am aware that there are significant penalties for submitting false information, SIGNATURE OF PRINCIPAL EXECUTIVE I including the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER YEAR MO DAY TYPED OR PRINTED CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

No closed mode operation. Veliger monitoring data is included as an attachment.

EPA Form 3320-1 (REV 3199) Previouseditions may be used Page I of 1

Mean # of Water Mean# of Water SUB NOTES: %

Sample Date ZM~m3 %Settlers Temp. ('C) Sample Date Asiatic Temp. (*C) LOCATION LOCATION Gravid Asiatic COLLECTED BY Clamslm3 Clam 12/07/2010 6 100 23 12/07/2010 0 23 1-25-545 PB 12/14/2010 0 0 10 12/14/2010 0 10 1-25-545 RS 12/22/2010 0 0 10.5 12/22/2010 0 10.5 1-ISV-24-1234 WE 12/29/2010 0 0 26 12/29/2010 0 26 1-25-545 WDT 01/04/2011 0 0 13 01/04/2010 0 13 1-25-545 PB 01/11/2011 0 0 22 01/11/2010 0 22 1-25-545 RS 01/18/2011 0 0 9.5 01/18/2010 0 9.5 1-ISV-24-1234 CR 01/25/2011 0 0 23 01/25/2011 0 23 1-25-545 WDT 02/02/2011 0 0 10 02/02/2011 0 10 1-25-545 PB 02/08/2011 0 0 9 02/08/2011 0 9 1-25-545 MJW 02/15/2011 0 0 23 02/15/2011 0 23 1-25-545 MLW 02/22/2011 20 100 10 02/22/2011 0 10 1-25-545 PB 03/01/2011 0 0 11 03/01/2011 0 11 1-ISV-24-1236 PB 03/08/2011 0 0 11 03/08/2011 0 11 1-ISV-24-1236 WE 03/16/2011 22 0 11 03/16/2011 0 11 1-ISV-24-1234 MLW 03/23/2011 0 0 11 03/23/2011 0 11 1-ISV-24-1234 MLW 03/30/2011 0 0 12 03/30/2011 0 12 1-ISV-24-1236 MLW 04/06/2011 18 100 15 04/06/2011 0 15 1-ISV-24-1234 HMW 04/08/2011 45 100 15.5 04/08/2011 0 15.5 1-1 SV-24-1236 WAW/PB 04/20/2011 21 100 16 04/20/2011 0 16 1-1 SV-24-1236 PB May 2011 No Samples Collected June 2011 No Samples Collected July 2011 No Samples Collected August 2011 No Samples Collected Sept 2011 No Samples Collected October 2011 No Samples Collected 11/20/2011 0 0 14 11/20/2011 18 14 1-ISV-24-1236 11/29/2011 0 0 14 11/29/2011 0 14 1-25-545 BC

PERMITTEE NAME/ADDRESS (Include Facility Name/Location if Different) NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

MAJOR Form Approved.

Name TVA - SEQUOYAH NUCLEAR PLANT DISCHARGE MONITORING REPORT (DMR)

Address (SUBR 01) OMB No. 2040-0004 P.O. BOX 2000 TN0026450 E ][101 T AINTfEROFFICE OPS-5N-SQdN) F - FINAL SODDY - DAISYTN. 37384 PERMIT NUMBER BIOMONITORING FOR OUTFALL 101

] DISCHARGE NUMBER]

Facilty TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY YEA M MONITORING PERIOD EFFLUENT YEAR MO DAY I IYEARI MO DAY ATTN: Brad Love Froml 111 11 1 01] Tol 111 11 3in NO DISCHARGE 11111 NOTE: Read instructions before completing this form.

PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO. FREQUENCY SAMPLE EX OF TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS IC25 STATRE 7DAY CHR SAMPLE ** >100.0 23 0 1 / 180 COMPOS CERIODAPHNIA MEASUREMENT TRP3B 1 0 PERMIT 43.2

PERCENT SEMI COMPOS EFFLUENT GROSS REQUIREMENT MINIMUM ANNUAL IC25 STATRE 7DAY CHR SAMPLE **

MEASUREMENT

>100.0 23 0 1/180 COMPOS PIMEPHALES TRP6C 1 0 PERMIT ** 43.2

PERCENT SEMI COMPOS EFFLUENT GROSS REQUIREMENT MIMINUM ANNUAL SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my direction or supervision in accordance with a system designed to assure that qualified personnel TELEPHONE DATE John T. Carlin properly gather persons who and evaluate the information submitted. Based on my inquiry of the person or manage the system, or those persons directly responsible for gathering the 0*ic President information, the information submitted is , to the best of my knowledge and belief, true, accurate, 423 843-7001 Sequoyah Site Vice President and complete. I am aware that there are significant penalties for submitting false information, 11 12 09 SIGNFIýE OF PRINCIPAL EXECUTIVE TYPED OR PRINTED ncluding the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER CODE YEAR MO DAY COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

Toxicity sampling was performed November 6 - 11. The report is included as an attachment.

EPA Form 3320-1 (REV 3199) Previouseditions may be used Page 1 of I

TENNESSEE VALLEY AUTHORITY TOXICITY TEST REPORT INTRODUCTION / EXECUTIVE

SUMMARY

Report Date: December 05, 2011

1. Facility / Discharger: Sequoyah Nuclear Plant / TVA

2. County / State: Hamilton / Tennessee

3. NPDES Permit #: TN0026450

4. Type of Facility: Nuclear-Fueled Electric Generating Plant

5. Design Flow (MGD): 1,579

6. Receiving Stream: Tennessee River (TRM 483.6)

7. 1Q10: 3491

8. Outfall Tested: 101

9. Dates Sampled: November 06 - 11, 2011

10. Average Flow on Days Sampled (MGD): 1685.2, 1686.5, 1680.1

11. Pertinent Site Conditions: Production / operation data will be provided upon request.

12. Test Dates: November 08 - 15, 2011

13. Test Type: Short-term Chronic Definitive

14. Test Species: Fathead Minnows (Pimephalespromelas)

Daphnids (Ceriodaphniadubia)

15. Concentrations Tested (%): Outfall 101: 10.8, 21.6, 43.2, 86.4, 100 Intake: 100 Pimephalespromelas: UV treated Outfall 101: 10.8, 21.6, 43.2, 86.4, 100 UV treated Intake: 100

16. Permit Limit Endpoint (%): Outfall 101: IC5 = 43.2%

17. Test Results: Outfall 101: Pimephalespromelas:.ICz> 100%

Ceriodaphniadubia: IC_5 > 100%

UV treated Outfall 101: Pimephalespromelas: IC?5 > 100%

Page 1 of 102

18. Facility

Contact:

Brad Love Phone #: (423) 843-6714

19. Consulting / Testing Lab: Environmental Testing Solutions, Inc.

20. Lab

Contact:

Jim Sumner Phone M: (828) 350-9364

21. TVA

Contact:

Donald W. Snodgrass Phone #: (256) 386-2787

22. Notes: Outfall 101 samples collected November 06.- 11,2011, showed no toxic effects to fathead minnows or daphnids. The resulting IC 25 values, for both species, were > 100 percent. Exposure of minnows and daphnids to intake samples resulted in no significant difference from the controls during this study period.

Fathead minnows were also exposed to UV treated Outfall 101 and intake samples since fish pathogens present in intake water have been the suspected cause of interference (anomalous dose response and high variability among replicates) in previous toxicity testing at Sequoyah. At the time this study was conducted, insignificant mortality occurred in minnows exposed to non-treated and UV treated samples.

Page 2 of 102

METHODS

SUMMARY

Samples:

1. Sampling Point: Outfall 101, Intake

2. Sample Type: Composite

3. Sample Information:

Date Date Date (MM-DD-YY) (MM-DD-YY) Arrival Initial (MM-DD-YY)

Sample Time (ET) Time (ET) Temp. TRC* Time (ET)

ID Collected Received (C (mg/L) Last Used By 101 11-06-11 0655to 11-07-11 1339 0.7,0.61 <0.10 11-08-11 1235 11-07-11 0555 11-09-11 1140 11-06-11 07l0 to 11-08-11 1235 11-07-11 1339 0.6 <0.10 11-09-11 1140 Intake 11-07-11 0610 110711 0610 11-09-11 1140 11-08-Il 0655 to 11-10-11 1148 11-09-11 1230 0.9, 0.7' <0.10 101 11-09-11 0555 11-11-11 1139 11-08-11 0705 to 11-10-11 1148 1139 11-09-11 1230 1.0 <0.10 11-11-11 Intake 11-09-11 0605 11-09-11 0605 11-11-11 1139 11-12-11 1149 11-10-11 O655to 101 11-11-11 0555 11-11-11 1220 2.1, 1.7't <0.10 11-13-11 1056 11-14-11 1142 11-12-11 1149 11-10-11 0705 to1-21114 Intake 0605 11-11-11 1220 2.0 <0.10 11-13-11 1056 11-11-11 0605 11-14-11 1142

TRC = Total Residual Chlorine tSamples were collected in two 2.5 gallon cubitainers. Temperature was measured in each cubitainer upon arrival.

ttSamples were collected in two 5 gallon cubitainers. Temperature was measured in each cubitainer upon arrival.

4. Sample Manipulation: Samples from Outfall 101 and intake were warmed to test temperature (25.0 +/- 1L.0C) in a warm water bath.

Aliquots of Outfall 101 and Intake samples were UV-treated through a 40-watt Smart UV Sterilizer (manufactured by Emperor Aquatics, Inc..) for 2 minutes.

Page 3 of 102

Pimephalespromelas Ceriodaphniadubia Test Organisms:

1. Source: Aquatox, Inc. In-house Cultures

2. Age: 20.33 - 20.58 hours6.712963e-4 days 0.0161 hours 9.589947e-5 weeks 2.2069e-5 months old < 24-hours old Test Method Summary:

1. Test Conditions: Static, Renewal Static, Renewal

2. Test Duration: 7 days Until at least 60% of control females have 3 broods

3. Control / Dilution Water: Moderately Hard Synthetic Moderately Hard Synthetic

4. Number of Replicates: 4 10

5. Organisms per Replicate: 10 1

6. Test Initiation: (Date/Time)

Outfall 101 11-08-11 1235 ET 11-08-11 1011ET UV Treated Outfall 101 11-08-11 1220ET

7. Test Termination: (Date/Time)

Outfall 101 11-15-11 1146ET 11-15-11 1006ET UV Treated Outfall 101 11-15-11 1130ET

8. Test Temperature: Outfall 101: Mean = 24.8°C Mean = 24.9'C (24.3 - 25.1 °C) (24.8 - 25.2 0C)

Test Temperature: UV-Treated Outfall 101: Mean = 24.8 0 C (24.4 - 25.2 0C)

9. Physical / Chemical Measurements: Alkalinity, hardness, total residual chlorine, and conductivity were measured at the laboratory in each 100% sample. Daily temperatures were measured in one replicate for each test concentration. Pre- and post-exposure test solutions were analyzed daily for pH and dissolved oxygen.

10. Statistics: Statistics were performed according to methods prescribed by EPA using ToxCalc version 5.0 statistical software (Tidepool Scientific Software, McKinneyville, CA).

Page 4 of 102

TOXICITY TEST RESULTS (see Appendix C for Bench Sheets)

1. Results of a Piniephalespromelas Chronic/ 7-day Toxicity Test.

(Genus species) (Type / Duration)

Conducted November 08 - 15, 2011 using effluent from Outfall 101.

Test Percent Surviving Solutions (time interval used - days)

(% Effluent) 1 2 3 [ 4 J 5 6 I 7 Control 100 100 100 100 100 100 100 10.8% 100 100 100 100 100 100 98 21.6% 100 100 100 100 100 100 100 43.2% 100 100 100 100 100 100 98 86.4% 100 100 100 100 100 100 100 100.0% 100 100 100 98 98 98 98 Intake 100 100 100 98 98 98 95 Test Solutions Mean Dry Weight (mg)

(% Effluent) 2(replicate number) u1 2 3 4 Mean Control 0.699 0.681 0.851 0.849 0.770 10.8% 0.781 0.774 0.737 0.673 0.741 21.6% 0.645 0.707 0.859 0.750 0.740 43.2% 0.720 0.757 0.616 0.717 0.703 86.4% 0.701 0.685 0.772 0.863 0.755 100.0% 0.767 0.800 0.793 0.684 0.761 Intake 0.713 0.695 0.733 0.706 0.712 IC 25 Value: > 100% Calculated TU Estimates: < 1.0 TUc*

Permit Limit: 43.2%

Permit Limit: 2.3 TUc 95% Confidence Limits:

Upper Limit: NA Lower Limit: NA

TUa = 100/LC5 0: TUc = 100/IC 25 Page 5 of 102

TOXICITY TEST RESULTS (see Appendix C for Bench Sheets)

2. Results of a Ceriodaphniadubia Chronic/ 7-day Toxicity Test.

(Genus species) (Type / Duration)

Conducted November 08 - 15, 2011 using effluent from Outfall 101.

Percent Surviving Test (time interval used - days)

Solutions 3 4 5 6 7

(% Effluent) 1 Control 100 100 100 100 100 100 100 10.8% 100 100 100 100 100 100 100 21.6% 100 100 100 100 100 100 100 43.2% 100 100 100 100 100 100 100 86.4% 100 100 100 100 100 100 100 100.0% 100 100 100 100 100 100 100 Reproduction (#young/female/7 days)

Test Solutions Data (replicate number)

(%Effluent) 1 ]2 3 [ 4 I5 J 6 77 8 1 9 [ 10 IMean Control 30 27 29 28 29 28 26 30 29 26 28.2 10.8% 32 32 30 31 30 35 31 32 33 31 31.7 21.6% 34 31 30 32 29 31 35 33 31 29 31.5 43.2% 34 38 29 34 32 33 30 35 36 35 33.6 86.4% 36 35 34 34 34 34 37 32 38 36 35.0 100.0% 34 35 39 36 34 36 35 35 33 34 35.1 IC2 5 Value: > 100% Calculated TU Estimates: < 1.0 TUc*

Permit Limit: 43.2%

Permit Limit: 2.3 TUc 95% Confidence Limits:

Upper Limit: NA Lower Limit: NA

TUa = 100/LCs0: TUc = 100/IC 25 Page 6 of 102

TOXICITY TEST RESULTS (see Appendix C for Bench Sheets)

2. Results of a Ceriodaphniadubia Chronic/ 7-day Toxicity Test.

(Genus species) (Type / Duration)

Conducted November 08 - 15, 2011 using water from Intake Percent Surviving Test (time interval used - days)

Solutions 1 2 3

(% Effluent) 1 2 6 7 Control 100 100 100 100 100 100 100 Intake 100 100 100 100 100 100 100 Reproduction (#young/female/7 days)

Test Solutions Data (replicate number)

(%Effluent) 1 2 3 4 5 6 7 8 910 Mean Control 27 27 27 27 30 29 28 26 27 28 27.6 Intake 32 36 32 36 32 33 32 31 34 34 33.2 IC 25 Value: > 100% Calculated TU Estimates: < 1.0 TUc*

Permit Limit: N/A Permit Limit: N/A 95% Confidence Limits:

Upper Limit: NA Lower Limit: NA

TUa = I00/LC50 : TUc = 100/IC 25 Page 7 of 102

TOXICITY TEST RESULTS, UV-TREATED (see Appendix C for Bench Sheets)

3. Results of a Pimephalespromelas Chronic/ 7-day Toxicity Test.

(Genus species) (Type / Duration)

Conducted November 08 - 15, 2011 using effluent from UV Treated Outfall 101.

Test Percent Surviving Solutions (time interval used - days)

(% Effluent) 1 ] 2 3 4 5 6 ] 7 Control 100 100 100 100 100 100 100 10.8% 100 100 100 100 100 100 100 21.6% 100 100 100 100 100 100 100 43.2% 100 100 100 100 100 100 100 86.4% 100 100 100 98 98 98 98 100.0% 100 100 100 100 100 100 100 Intake 100 100 100 100 100 100 100 Mean Dry Weight (mg)

(%Eflen)

(%replicate 1 ] 2 3 number)

] 4 1] Mean Control 0.681 0.718 0.733 0.762 0.724 10.8% 0.688 0.626 0.647 0.726 0.672 21.6% 0.655 0.729 0.756 0.672 0.703 43.2% 0.706 0.739 0.682 0.757 0.721 86.4% 0.621 0.648 0.794 0.597 0.665 100.0% 0.835 0.676 0.665 0.681 0.714 Intake 0.744 0.821 0.774 0.706 0.761 IC 25 Value: > 100% Calculated TU Estimates: < 1.0 TUc*

95% Confidence Limits:

Upper Limit: NA Lower Limit: NA

TUa = l00/LC5 0: TUc = 100/IC2 5 REFERENCE TOXICANT TEST RESULTS (see Appendix A and D)

Species Date Time [Duration Toxicant Results (IC 25)

Pimephalespromelas November 08 - 15, 2011 1245 7-days KC1 0.78 g/L Ceriodaphniadubia November 08 - 15, 2011 1000 7-days NaCI 1.05 g/L Page 8of 102

PHYSICAL/CHEM ICAL

SUMMARY

Water Chemistry Mean Values and Ranges fbr Pimephales promnelas and Ceriodap/miadubia Tests, Non-treated Sequoyah Nuclear Plant (SQN) Outfall 101 performed November 08-15, 2011.

0 Test Sample ID Temperature CC) Dissolved Oxygen (mg/L) pH (S.U.) Conductance Alkalinity Hardness Total Residual Initial Final Initial Final Initial Final (Amhos/cm) (mgJL CaCO3) (mgfL CaCO 3) Chlorine (nig/L) 7.7 7.5 8.02 7.80 311 62 89 Control 24.8 24.6 24.7 - 24.9 24.3 - 24.8 7.6 - 7.8 7.0 - 7.7 7.85 - 8.12 7.73 - 7.92 301 - 319. 61 - 62 87 - 91 7.7 7.4 7.99 7.77 297 10.8% 24.9 24.6 24.7 - 25.0 24.3 - 24.9 7.6 - 7.9 6.8 - 7.7 7.87 - 8.07 7.66 - 7.88 293 - 303 24.9 24.6 7.7 7.4 8.00 7.77 283 24.7 - 25.0 24.5 - 24.8 7.6 - 7.8 6.9 - 7.7 7.87 - 8.10 7.68 - 7.86 278 - 286 4.

43.2%/ 24.9 24.7 7.7 7.4 7.99 7.79 259 4 24.7 - 25.0 24.5 - 24.8 7.6 - 7.8 6.9 - 7.7 7.88 - 8.10 7.66 - 7.89 256 - 262

6. 25.0 24.6 7.7 7.4 7.97 7.82 205 11 86.4%

24.8 - 25.1 24.5 - 24.8 7.6 - 7.8 6.7 - 7.7 7.84 - 8.09 7.70 - 7.95 201'- 210 7.8 7.4 7.94 7.85 188 73 77 < 0.10 100.0% 25.0 24.7 24.8 - 25.1 24.5 - 24.8 7.7 - 7.9 6.8 - 7.7 7.75 - 8.06 7.79 - 7.97 186 - 190 73 - 73 77 - 77 <0.10- <0.10 7.9 7.5 7.98 7.87 187 73 77 < 0.10 Intake 24.9 24.6 24.8 - 25.0 24.5 - 24.7 7.7 - 8.2 7.0 - 7.8 7.87 - 8.08 7.80 - 7.96 186 - 189 73 - 73 75 - 79 <0.10- <0.10 7.7 7.7 8.02 7.97 311 62 89 Control 24.8 25.0 24.8 - 24.9 24.8 - 25.2 7.6 - 7.8 7.6 - 8.0 7.85 - 8.12 7.87 - 8.02 301 - 319 61 - 62 87 - 91 0.% 24.9 24.9 7.7 7.7 7.99 7.97 297 24.8 - 25.0 24.8 - 25.2 7.6 - 7.9 7.5 - 7.9 7.87 - 8.07 7.87 - 8.03 293 - 303

. 26 24.9 25.0 7.7 7.7 8.00 7.98 283 24.8 - 25.0 24.8 - 25.2 7.6 - 7.8 7.6 -- 7.9 7.87 - 8.10 7.88 - 8.02 278 - 286 z 41 43.2% 24.9 25.0 7.7 7.8 7.99 7.99 259

,X 24.8 - 25.0 24.8 - 25.2 7.6 - 7.8 7.6 - 7.9 7.88 - 8.10 7.89 - 8.05 256 - 262

86.4% 25.0 24.9 7.7 7.8 7.97 8.03 205 24.9 - 25.0 24.8 - 25.1 7.6 - 7.8 7.6 - 8.0 7.84 - 8.09 7.94 - 8.09 201 - 210 24.9 7.8 7.8 7.94 8.04 188 73 77 < 0.10 100.0% 25.0 24.8 - 25.1 24.8 - 25.1 7.7 - 7.9 7.7 - 8.0 7.75 - 8.06 7.93 - 8.09 186 - 190 73 - 73 77 - 77 <0.10- <0.10 7.9 7.8 7.98 8.08 187 73 77 < 0. 10 Intake 25.0 24.9 24.8 - 25.1 24.8 - 25.0 7.7 - 8.2 7.7 - 8.0 7.87 - 8.08 7.95 - 8.22 186 - 189 73 - 73 75 - 79 <0.10- <0.10 Overall temperature (CC) Average Minimum Maximum Pimnephalespromnelas 24.8 24.3 25.1 Ceriodaphniadubia 24.9 24.8 25.2

PHYSICALJCHEMICAL

SUMMARY

(D Water Chemistry Mean Values and Ranges for PimephalespromelasTest, UV-treated Sequoyah Nuclear Plant (SQN) Outfall 101 performed November 08-15, 2011.

NJ 0 Test Sample ID Temperature (C) Dissolved Oxygen (mg/L) pH (S.U.) Conductance Alkalinity Hardness Total Residual Initial Final Initial Final Initial Final (pmhos/cm) (mg/L CaCO 3 ) (mg/L CaCO3 ) Chlorine (mg/L) 7.5 7.99 7.76 310 61 88 Control 24.9 24.6 7.8 24.6 - 25.0 24.5 - 24.8 7.6 - 8.0 6.8 - 7.8 7.87 - 8.06 7.65 - 7.88 305 - 316 60 - 61 87 - 89 7.8 7.4 7.99 7.78 296 10.8% 24.9 24.7 24.7 - 25.0 24.5 - 24.9 7.7 - 8.0 6.8 - 7.8 7.89 - 8.07 7.70 - 7.87 290 - 305

1.6% 24.9 24.7 7.8 7.5 7.98 7.81 285 24.8 - 25.0 24.5 - 24.9 7.7 - 8.0 7.0 - 7.8 7.89 - 8.06 7.73 - 7.91 281 - 290 Q. 25.0 24.7 7.8 7.5 7.98 7.80 260 43.2%

24.8 - 25.1 24.5 - 25.0 7.6 - 8.1 6.9 - 7.8 7.88 - 8.06 7.70 - 7.89 258 - 262 86.4% 25.0 24.6 7.8 7.5 7.97 7.86 206 24.8 - 25.1 24.4 - 24.8 7.6 - 8.1 6.9 - 7.8 7.87 - 8.05 7.75 - 7.95 200 - 211 7.94 7.84 190 73 77 <0.10 100.0% 25.1 24.6 7.9 7.5 24.9 - 25.2 24.5 - 24.9 7.6 - 8.0 7.0 - 7.8 7.82 - 8.04 7.76 - 7.93 189 - 192 73 - 73 75 - 79 <0.10 - <0.10 7.98 7.86 187 73 77 < 0.10 Intake 25.0 24.6 7.9 7.5 24.9 - 25.2 24.4 - 24.9 7.7 - 8.2 7.1 - 7.8 7.86 - 8.06 7.76 - 7.97 184 - 191 71 - 75 75 - 79 <0.10 - <0.10 Ave rage Maximum Overall temperature (C) Minimum Pimephalesproinelas 24.8 24.4 25.2

SUMMARY

/ CONCLUSIONS Outfall 101 samples collected November 06 - 11, 2011, showed no toxic effects to fathead minnows or daphnids. The resulting IC25 values, for both species, were >

100 percent. Exposure of minnows and daphnids to intake samples resulted in no significant difference from the controls during this study period.

Fathead minnows were also exposed to UV treated Outfall 101 and intake samples since fish pathogens present in intake water have been the suspected cause of interference (anomalous dose response and high variability among replicates) in previous toxicity testing at Sequoyah. At the time this study was conducted, insignificant mortality occurred in minnows exposed to non-treated and UV treated samples.

Page 11 of 102

Appendix A ADDITIONAL TOXICITY TEST INFORMATION

SUMMARY

OF METHODS

1. Pitnephalespromelas Tests were conducted according to EPA-821 -R-02-013 (October 2002) using four replicates, each containing ten test organisms, per treatment. Test vessels consisted of 500-mL plastic disposable cups, each containing 250-mL of test solution.

2. Ceriodaphniadubia Tests were conducted according to EPA-82 1-R-02-013 (October 2002) using ten replicates, each containing one test organism, per treatment. Test vessels consisted of 30-mL polypropylene cups, each containing 15-mL of test solution.

DEVIATIONS / MODIFICATIONS TO TEST PROTOCOL

1. Pimephalespromelas None

2. Ceriodaphniadubia None DEVIATIONS / MODIFICATIONS TO PRETEST CULTURE OR HOLDING OF TEST ORGANISMS

1. Pimephalespromelas None

2. Ceriodaphniadubia None Page 12 of 102

PHYSICAL AND CHEMICAL METHODS

1. Reagents, Titrants, Buffers, etc.: All chemicals were certified products used before expiration dates (where applicable).

2. Instruments: All identification, service, and calibration information pertaining to laboratory instruments is recorded in calibration and maintenance logbooks.

3. Temperature was measured by SM 2550 B.

4. Dissolved oxygen was measured by SM 4500 0 G.

5. The pH was measured by SM 4500 H+ B.

6. Conductance was measured by SM 22510 B.

7. Alkalinity was measured by SM 2320 B.

8. Total hardness was measured by SM 2340 C.

9. Total residual chlorine was measured by ORION Electrode Method 97-70.

QUALITY ASSURANCE Toxicity Test Methods: All phases of the study including, but not limited to, sample collection, handling and storage, glassware preparation, test organism culturing/acquisition and acclimation, test organism handling during test, and maintaining appropriate test conditions were conducted according to the protocol as described in this report and EPA-82 I-R-02-013. Any known deviations were noted during the study and are reported herein.

REFERENCE TOXICANT TESTS (See Appendix D for control chart information)

1. Test Type: 7-day chronic tests with results expressed as IC 25 values in g/L KC1 or NaCl.

2. Standard Toxicant: Potassium Chloride (KC1 crystalline) for Pimephalespromelas.

Sodium Chloride (NaCI crystalline) for Ceriodaphniadubia.

3. Dilution Water Used: Moderately hard synthetic water.

4. Statistics: ToxCalc software Version 5.0 was used for statistical analyses.

Page 13 of 102

REFERENCES

1. NPDES Permit No. TN0026450.

2. USEPA. Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, EPA-82 1-R-02-013 (October 2002).

3. Standard Methods for the Examination of Water and Wastewater, 2 1st Edition, 2005.

4. Quality Assurance Program: Standard Operating Procedures, Environmental Testing Solutions, Inc (most current version).

Page 14 of 102

Sequoyah Nuclear Plant Biomonitoring November 08 - 15, 2011 Appendix B Diffuser Discharge Concentrations of Total Residual Chlorine, Diffuser Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion and Mollusks-During Toxicity Test Sampling Page 15 of 102

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test Sampling, March 12, 1998- November 11, 2011 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat- H-130M Hypochlorite mg/L mg/L mg/L mg/L PF mg/L mg/L TRC Phosphate Copolymer DMAD mg/L Quat TRC Azole 03/12/1998 0.016 -

03/13/1998 0.015 -

03/14/1998 0.013 -

03/15/1998 0.030 -

03/16/1998 0.013 -

03/17/1998 0.020 -

03/18/1998 0.018 09/08/1998 0.015 - 0.014 0.005 - - 0.021 0.003 - 0.031 0.011 - - -

09/09/1998 0.014 - 0.060 0.021 - - -

09/10/1998 0.013 - 0.055 0.019 - - -

09/11/1998

< 0.001 - 0.044 0.015 - - -

09/12/1998

< 0.001 - 0.044 0.015 - - -

09/13/1998 09/14/1998 0.008 0.044 0.015 -

02/22/1999 < 0.001 - - -

02/23/1999 0.005 02/24/1999 0.009 02/25/1999 0.012 02/26/1999 0.008 02/27/1999 < 0.001 02/28/1999 < 0.001 08/18/1999 - 0.015 0.069 0.024 0.006 08/19/1999 - 0.012 0.068 0.024 - -

08/20/1999 - 0.023 0.070 0.024 - 0.120 0.022 0.068 0.024 - -

08/21/1999 -

08/22/1999 - 0.022 .0.068 0.024 -

08/23/1999 - 0.025 0.068 0.024 0.006 08/24/1999 - 0.016 0.067 0.023 0.020 1 Page 16 of 102

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test Sampling, March 12, 1998- November 11, 2011 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat- H-130M Hypochlorite mg/L mg/L mg/L mg/L PF mg/L mg/L TRC Phosphate Copolymer DMAD mg/L Quat TRC Azole 01/31/2000 -< 0.002 0.026 0.009 02/01/2000 - 0.011 0.026 0.028 -

02/02/2000 - 0.028 0.026 0.009 0.006 02/03/2000 - 0.008 0.027 0.009 - -

02/04/2000 - 0.006 0.027 0.009 0.005 0.109 02/05/2000 -< 0.002 0.027 0.009 - - -

02/06/20001 - < 0.002 0.027 0.009 - - -

07/26/2000 -< 0.0057 0.055 0.019 - - -

07/27/2000 - 0.019 0.055 0.019 - - -

07/28/2000 - 0.0088 0.053 0.018 0.004 0.108 -

07/29/2000 -< 0.0088 0.055 0.019 - - -

07/30/2000 -< 0.0076 0.055 0.019 -

07/31/2000 -< 0.0152 0.055 0.019 0.006 08/01/2000 - < 0.0141 0.055 0.019 0.005 12/11/2000 - 0.0143 0.025 0.020 0.005 12/12/2000 - 0.0092 0.025 0.020 0.005 12/13/2000 -< 0.0120 0.025 0.020 -

12/14/2000 -< 0.0087 0.025 0.020 -

12/15/2000 - 0.0120 0.025 0.020 0.005 12/16/2000 < 0.0036 0.025 0.020-0.020

- < 0.0036 0.025 12/17/2000 08/26/2001 - 0.017 0.06 0.021 0.006 08/27/2001 - <0.0096 0.06 0.021 0.005 0.021 08/28/2001 - <0.0085 0.06 0.021 - -

08/29/2001 - <0.0094 0.059 0.020 0.005 0.021 08/30/2001 - <0.0123 0.06 0.021 0.005 -

08/31/2001 - <0.005 0.059 0.020 11/25/2001 - <0.0044 - - -

11/26/2001 - <0.0119 0.024 0.02 0.005 - -

11/27/2001 - 0.0137 0.023 0.019 0.007 - -

11/28/2001 - <0.0089 0.022 0.019 0.006 - -

11/29/2001 - 0.0132 0.024 0.02 0.007 - -

11/30/2001 - < 0.0043 0.024 0.02. - - -

12/09/2001 - <0.0042 - -

12/10/2001 - <0.0042 12/11/2001 - <0.0104 - - -

12/12/2001 - 0.0128 0.024 0.02 0.008 12/13/2001 - <0.0088 0.024 0.02 -

12/14/2001 - 0.0134 0.024 0.02 0.007 Page 17of 102

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test Sampling, March 12, 1998 - November 11, 2011 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat- H-130M Hypochlorite mg/L mgIL mg/L mg/L PF mg/L mg/L TRC Phosphate Copolymer DMAD mg/L Quat TRC Azole 01/02/2002 < 0.0079 0.023 0.02 0.006 01/03/2002 -< 0.0042 0.023 0.014 -

01/04/2002 - 0.0124 0.024 0.014 0.009

-< 0.0042 - - -

01/05/2002

-< 0.0042 - - -

01/06/2002 01/07/2002 - < 0.0089 0.024 0.014 0.006 -

02/24/2002 -< 0.004 - - -

02/25/2002 -< 0.004 0.023 0.023 --

02/26/2002 - 0.0143 0.023 0.023 0.007 02/27/2002 -< 0.0041 0.023 0.023 -

02/28/2002 -< 0.0041 0.024 0.008 03/01/2002 -< 0.0041 0.024 0.008 05/05/2002 05/06/2002 - - 0.058 0.02 0.014 05/07/2002 - - 0.058 0.02 0.015 05/08/2002 - - 0.056 0.019 -

05/09/2002 - - 0.057 0.02 0.014 05/10/2002 - - 0.056 0.019 08/04/2002 - <0.0058 - -

08/05/2002 - <0.0058 0.053 0.018 0.025 08/06/2002 - 0.0092 0.053 0.018 - -

08/07/2002 - <0.0107 0.055 0.019 0.007 08/08/2002 - <0.0061 0.055 0.019 -

08/09/2002 - 0.0152 0.054 0.018 0.008 10/06/2002 - <0.00497 - --

10/07/2002 - 0.0153 0.054 0.018 0.009 10/08/2002 - <0.0092 0.054 0.018 0.007 10/09/2002 - 0.0124 0.053 0.018 0.009 10/10/2002 - 0.0134 0.054 0.018 0.009 10/11/2002 - <0.0042 0.054 0.018

<0.0035 - - - - -

01/12/2003 -

01/13/2003 - <0.006 0.025 0.019 0.009 - -

01/14/2003 - <0.0118 0.026 0.020 - -

01/15/2003 - <0.0063 0.026 0.020 0.009 - -

01/16/2003 - <0.0034 0.026 0.020 - -

01/17/2003 - <0.0034 0.026 0.009 -

04/06/2003 - <0.0073 - -

04/07/2003 - <0.0189 - 0.021 04/08/2003 - <0.0117 - 0.021 -

04/09/2003 - <0.0139 - 0.021 0.016 04/10/2003 - <0.0113 - 0.021 0.018 04/11/2003 - <0.0073 - 0.022 - I I Page 18 of 102

Table B-1 (continued). Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Growth of Microbiologically Induced Bacteria and Mollusks, During Toxicity Test Sampling, March 12, 1998- November 11, 2011 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat- H-130M Hypochlorite mg/L mg/L mg/L mg/L PF mg/L mg/L TRC Phosphate Copolymer DMAD mg/L Quat TRC Azole 06/15/2003 - < 0.0045 - - - - -

06/16/2003 - < 0.0037 0.057 0.020 - - 0.022 06/17/2003 - < 0.0048 0.041 0.014 - - 0.024 06/18/2003 - < 0.0048 0.041 0.014 - - 0.024 06/19/2003 - < 0.0085 0.058 0.020 - - 0.025 06/20/2003 < 0.0048 0.058 0.020 - - 0.025

<0.0050 - - - -

08/03/2003

<0.0050 0.058 0.020 - -

08/04/2003 08/05/2003 <0.0051 0.057 0.020 - - 0.025 08/06/2003 <0.0084 0.057 0.020 - - 0.025 08/07/2003 0.0129 0.057 0.020 - - 0.024 08/08/2003 0.0153 0.057 0.020 0.009 - -

10/05/2003 <0.0043 0.057 0.020 - - -

10/06/2003 <0.0043 0.057 0.020 - - 0.025 10/07/2003 <0.0090 0.057 0.020 - - 0.025 10/08/2003 <0.0106 0.057 0.020 - - 0.025 10/09/2003 0.0181 0.026 0.022 - - 0.025 10/10/20031 0.0183 0.026 0.024 0.009 - -

02/01/2004 0.0093 0.027 0.009 --

02/02/2004 <0.0034 0.026 0.009 -

02/03/2004 <0.0034 0.026 0.009 -

02/04/2004 0.0124 0.026 0.009 0.009 02/05/2004 <0.0034 0.026 0.009 - --

02/06/2004 0.0105 0.026 0.009 0.010 - -

05/04/2004 <0.0123 0.026 0.019 - - 0.025 05/05/2004 <0.0144 0.026 0.014 0.009 - 0.025 05/06/2004 <0.0146 0.037 0.013 - - 0.025 05/07/2004 0.0227 0.058 0.020 0.009 - 0.025 05/08/2004 0.016 0.060 0.021 - - -

05/09/2004 <0.0104 0.058 0.020 07/04/2004 0.0217 0.057 0.019 - --

07/05/2004 <0.0085 0.057 0.020 0.009 - -

07/06/2004 <0.0077 0.058 0.020 - - 0.031 07/07/2004 0.0252 0.056 0.019 - - 0.031 07/08/2004 0.0223 0.057 0.019 0.009 - -

07/09/2004 0.0182 0.057 0.020 0.009 - -

Page 19 of 102

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test Sampling, March 12, 1998 - November 11, 2011 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-PF H-130M Nalco H-150M Hypochlorit mg/L mg/L mg/L mg/L mg/L mg/L 73551 mg/L e TRC Phosphate Copolymer DMAD Azole Quat mg/L Quat mg/L EO/PO TRC 11/07/2004 - <0.0187 0.000 0.014 - -

11/08/2004 - <0.0192 0.047 0.030 - - -

11/09/2004 - <0.0233 0.048 0.016 - - 0.041 11/10/2004 - <0.0149 0.047 0.016 - - 0.041 11/11/2004 - <0.0149 0.049 0.017 - - 0.043 11/12/2004 - <0.0253 0.048 0.017 - 0.042 02/06/2005 - <0.0042 0.028 0.010 -

02/07/2005 - <0.0116 0.028 0.010 0.007 02/08/2005 - <0.0080 0.028 0.010 -

02/09/2005 - 0.0199 0.028 0.010 02/10/2005 - <0.0042 0.028 0.010 02/11/2005 0.0155 0.028 0.010 0.007 06/05/2005 0.0063 -

06/06/2005 0.0043 -- 0.037 06/07/2005 0.0103 -- 0.037 06/08/2005 0.0295 -- 0.037 06/09/2005 0.0129 - - -

06/10/2005 0.0184 - -

07/17/2005 0.0109 0.026 0.009 -

07/18/2005 0.0150 0.026 0.009 0.036 07/19/2005 0.0163 0.026 0.009 - 0.036 07/20/2005 0.0209 0.026 0.009 0.014 0.036 07/21/2005 0.0242 0.026 0.009 - -

07/22/2005 0.0238 0.054 0.018 0.014 10/30/2005 0.0068 - - -

10/31/2005 0.0112 - - -

11/01/2005 0.0104 -- 0.035 11/02/2005 0.0104 -- 0.036 11/03/2005 0.0117 -- 0.036 11/04/2005 0.0165 - 0.035 11/14/2005 - 0.0274 - - - - - -

11/15/2005 - 0.0256 - - - - - - -

11/16/2005 - 0.0234 - - - - - - -

11/17/2005 - 0.0231 - - - - - - -

11/18/2005 - 0.0200 - - - - - - -

11/19/2005 - 0.0116 - - - - - - -

Page 20 of 102

Table B-1 (continued). Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Growth of Microbiologically Induced Bacteria and Mollusks, During Toxicity Test Sampling, March 12, 1998 - November 11, 2011 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-PF H-130M Nalco H-150M MSW Hypochlorite mg/L mg/L mg/L mg/L mg/L mg/L 73551 mg/L 101 mg/L TRC Phosphate Copolymer DMAD Azole Quat mg/L Quat mg/L TRC EO/PO Phosphate 11/12/2006 - 0.0055 - - - - - - -

11/13/2006 - 0.0068 - - - - - 0.037 -

11/14/2006 - 0.0143 - - - - - 0.037 -

11/15/2006 - 0.0068 - - - - - 0.037 -

11/16/2006 - 0.0267 - - - - - 0.037 -

11/17/2006 - 0.0222 - - - - - - -

11/26/2006 - 0.0188 - - - - - - -

11/27/2006 - 0.0138 - - - - - - -

11/28/2006 - 0.0120 - - - - - - -

11/29/2006 - 0.0288 - - - - - - -

11/30/2006 - 0.0376 - - - - - - -

12/01/2006 - 0.0187 - - - - - - -

05/28/07 - -- - 0.015 05/29/07 - - - 0.036 0.015 05/30/07 - 0.0084 - - 0.017 0.036 0.015 05/31/07 - 0.0103 - - 0.036 0.015 06/01/07 - 0.0164 - - - - - 0.017 0.036 0.015 06/02/07 - 0.0305 - - - - - - - 0.015 12/02/07 - 0.0241 - - - - - -

12/03/07 - 0.0128 - - - - - - -

12/04/07 - 0.0238 - - - - - - -

12/05/07 - 0.0158 - - - - - - - -

12/06/07 - 0.0162 - - - - - - - -

12/07/07 - 0.0175 - - - - - - -

04/13/08 - 0.0039 - - - - - - - -

04/14/08 - 0.0124 - - - - - - - -

04/15/08 - 0.0229 - - - - - - - -

04/16/08 - 0.0143 - - - - - - -

04/17/08 - 0.0120 - - - - - - -

04/18/08 - 0.0149 -

10/26/08 - 0.0260 - -

10/27/08 - 0.0151 - 0.017 -

10/28/08 - 0.0172 - - 0.041 -

10/29/08 - 0.0154 - 0.018 0.041 0.030 10/30/08 - - - - 0.041 0.030 10/31/08 - 0.0086 - 0.041 0.030 Page 21 of 102

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test Sampling, March 12, 1998 - November 11, 2011 Date Sodium Towerbrorr PCL- PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSW Hyochlorite mg/L 222 mg/L mg/L -PF mg/L mg/L 73551 CT1300 mg/L 101 mg/L TRC mg/L Copolymer DMAD Azole Quat mg/L mg/L Quat mg/L TRC Phosph EO/PO Quat Phosphate ate 02/08/09 - 0.0197 - 0.017 -

02/09/09 - 0.0237 - 0.017 - - -

02/10/09 - 0.0104 - 0.021 - - -

02/11/09 - 0.0155 - 0.017 - - -

02/12/09 - 0.0106 - 0.017 - - -

02/13/09 - - -

05/10/09 - 0.0129 - -

05/11/09 - 0.0415 - 0.0446 -

05/12/09 - 0.0053 - 0.0396 -

05/13/09 - 0.0049 - 0.0396 -

05/14/09 - <0.0141 - 0.0397 -

05/15/09 <0.0160 11/15/09 0.025 11/16/09 0.0152 11/17/09 0.0255 - -

11/18/09 0.0306 - -

11/19/09 0.0204 - -

11/20/09 0.0093 - -

05/09/10 0.0192 05/10/10 0.0055 05/11/10 0.0100 - - 0.039 05/12/01 0.0171 - - 0.039 05/13/10 0.0041 - - 0.039 05/14/10 0.0099 - - 0.039 Page 22 of 102

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test Sampling, March 12, 1998- November 11, 2011 Date Sodium Towerbronr PCL- PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSW Hypochlorite mg/L 222 mg/L mg/L -PF mg/L mg/L 73551 CT1300 mg/L 101 mg/L TRC mg/L Copolymer DMAD Azole Quat mg/L mg/L Quat mg/L TRC Phosph EO/PO Quat Phosphate ate 10/31/10 - - - -

11/01/10 - 0.0122 - - - -

11/02/10 - 0.0112 - - - - -

11/03/10 - 0.0163 - - - -

11/04/10 - 0.0107 - - - -

11/05/10 - 0.0132 - - - -

05/01/2011 - -

05/02/2011 - 0.04 - -

05/03/2011 - - 0.04 - -

05/04/2011 - 0.0155 0.04 - -

05/05/2011 - 0.0179 0.04 - -

05/06/2011 - 0.0089 - - -

11/06/2011 - 0.0168 11/07/2011 - 0.0225 11/08/2011 - 0.0141 11/09/2011 - 0.0239 11/10/2011 - 0.0242 11/11/2011 - 0.0231 Page 23 of 102

Sequoyah Nuclear Plant Biomonitoring November 08 - 15, 2011 Appendix C Chain of Custody Records and Toxicity Test Bench Sheets Page 24 of 102

1 .1 BIOMONITORING CHAIN OF CUSTODY RECORD Page _1 of 1 Plient: TVA Environmental Testing Solution, Inc. Delivered By (Circle One):

roject Name: Sequoyah NP Toxicity 351 Depot Street. FedEx UPS Bus Client Or 11.0. Number: N/A Asheville, NC Other (specify):

General Comments:

Facility Sampled: Sequoyah NP 28801 , A o I,-,/--

- go .

NPDESNumber:TN0026450 Phone: 828-350-9364 VV CID, r: lov/ 146-I0 1 5,Q0 Collected By: Dustin Bipegarf tv: 11IV£ Fax: 828-350-9368 Fil dniiainNumber & Flow *.: ,,::.*.,:,.,~*g*. .*o,,,..,z...

Collection DateTime Volume (MGD)R F d DentictionVolle prab/Comp. (Mark as Appropriate) - .'-.

4 lieif Yate Yes, No Trace 1Sg. wyen SQN-TNT-TOX Comp f, 4/7//7/0o "(2.5gal) If//- A Sample Custody - Fill In From Top Down r u.- t_ pl!.i

& '. n1 Relinquished By (Signature): Date/Time Received By (Signature): Date/Time

___ ___ __ __ _ _ _ _ _ _ _ 0I/%a-ii7/ ~W ______________E____

Instructions: Clients should fill in all areas except those in the "Laboratory Use" block. Biomonitoring samples are preserved by storing them at 60C and shipping them in ice. The hold time for each sample is 36 hours4.166667e-4 days 0.01 hours 5.952381e-5 weeks 1.3698e-5 months from the time of collection. Therefore, please collect and ship in such a way that the laboratory will receive the samples with ample time to initiate testing within that time frame. Samples shipped overnight on Friday via FedEx or UPS must be marked for Saturday delivery or they will not arrive until the following Monday.

II I:ETS 4'.Wue Whole Effluent Toxicity Sample Receipt Log-

Sample temperature performed using Sample Receiving Thermometer: SN 6338.

Date Time Received Received Sample Project Sample _ 1, Sample name and description State Comments from temperature (*C) noumber received received by to 11-07-11 1339 JSuner TVACourier 0.7/0.6 7477 111107.01 TVA- uah Nuclear Plant - Outfall 101 TN 11-07-11 1339 i. SumneEr TVA Courier 0.6 7477 111107.02 TVA-S h Nuclear Plant - Intake TN to SOP G4 - Exhibit G4.2, revision 06-29-09

BIOMONITORING CHAIN OF CUSTODY RECORD Page _1 of 1 lient: TVA Environmental Testing Solution, Inc. Delivered By (Circle One):

lfoject Name: Sequoyah NP Toxicity 351 Depot Street. FedEx UPS Bus Client Crier 4). Number: N/A Asheville, NC Other (specify):._

ca General Comments:

Nicility Sampled: Sequoyah NP 28801 Gnr Comm :0430 NPDES Number: TN0026450 Phone: 828-350-9364 Collected By: Dust inegar/ W, Fax: 828-350-9368 Number& Flow Field Identification ~' Grab/Comp. Collection Date/Time Voumber & (MGRinEvnt Sample DescriptionCollume (MGD) (Mark as Appropriate) Laboratory Use Date Time Yes If'Yes, No Trace EITS Logý ,AnivalTemp. 'By. Timeý Appear-Inches _ _ Number ~ CC) mice SQN-101-TOX Comp E- osOr- 2(2.5gal) -"/R SQN-INT-TOX Comp 06-.r 1(2.5 gal) ff//9i Sample Custody - Fill In From Top Down ,,It 5S*-*AS (t1T.- S1A" ftL t*L.L*AW-b:

%a T~~ rVMLr.. ' . -9AM-Relinquished By (Signature): Date/Time Received By (Signature): Date/Time s"*94 ,M

\A/0 /// 69-.

ir Instructions: Clients should fill in all areas except those in the "Laboratory Use" block. Biomonitoring samples are preserved by storing them at 61C and shipping them in ice. The hold time for each sample is 36 hours4.166667e-4 days 0.01 hours 5.952381e-5 weeks 1.3698e-5 months from the time of collection. Therefore, please collect and ship i.n such a way that the laboratory will receive the samples with ample time to initiate testing within that time frame.. Samples shipped overnight on Friday via FedEx or UPS must be marked for Saturday delivery or they will not arrive until the following Monday.

N)o-6 TETS Whole Effluent Toxicity Sample Receipt Log

Sample temperature performed using Sample Receiving Thermometer: SN 6338.

Date Time Received Received Sample Project Sample Sample name and description SLate Comments received received by fronm temperature (*C)* nomber 11-09-11 0945 K. Keenan UPS 0.6 7490 111109.01 Microbac - Rockin ham WWTP NC 11-09-11 0945 K. Keenan -UPS 0.6 7491 111109.02 Microbac - Roseboro WWrP NC 11-09-11 0945 K. Keenan UPS 0.6 7492 111109.03 Microbac - J.P. Stevens - Wagram WWTP NC 11-09-11 0945 K. Keenan UPS 0.5 7493 111109.04 Enviro Chemists - Manteo WWTP NC

.m 11-09-11 0945 K. Keenan UPS 0.8 7494 111109.05 Atlantic Beach WWTP NC 11-09-11 0945 K. Keenan UPS 0.8 7495 111109.06 MoreheadCity WWTP NC 11-09-11 1007 K. Keenan Fed - Ex 1.0 7496 111109.07 Bladenboro WWTP NC S1I-9-11 1007 K. Keenan Fed- E. 0.7 7497 111109.08 Progress Energy Carolinas - Cape Fear SE. NC 11-09-11 1007 K. Keenan Fed- Ex 0.7 7497 111109.09 Progress Ener Carolinas - Cape Fear S. E. Upstream/Intake NC 11-09-11 1007 K. Keenan Fed- Ex 0.7 7498 111109.10 City ofGastonia- Dallas WWTP NC 11-09-11 1007 K. Keenan Fed - Ex 0.6 7499 111109.11 United Water - Enfield WWTP NC 11-09-11 1007 K. Keenan Fed-Ex 1.7 7500 111109.12 ProgressEnergy Carolinas - Lee Steam Plant NC 11-09-I1 1007 K. Keenan Fed - Ex 0.8 7501 111109.13 Duke Energy Cu *rtlion - Marshall Steam Station NC 1-09-1 i 1007 K. Keenan Fed - Ex 0.6 7502 111109.14 United Water- Scotland Neck WWTP NC 11-09-11 1007 K Keenan Fed - Ex 0.9 7503 111109.15 Progress Energy Carolinas -Mayo Steam Electric Plant NC 11-09-11 1007 K. Keenan Fed - Ex 0.5 7504 111109.16 ProresshEnergy Carolirs -SharonHarrisPlntC 11-09-11 1007 K. Keenan Fed - EX 0.4 7505 111109.17 Washington WWTP NC 11-09-11 1007 K. Keecan Fed - Es 0.4 7506 111109.18 Caolina Beach WWTP NC NC 0.5 7507 111109.19 WashiPgtoneWrP ShearonHarrisPlantNC S11-09-11 1007 K. Keenan Fed - Ex 1-09-lI 1132 K. Keenan Dash Courier 0.8 7508 111109.20 OWASA - Mason Farm WWTP NC 11-09-LI 1230 J. Sumner TVACouricr 0.9/0.7 7477 111109.21 TVA-Sequoyah NuclearPlant-Outfall 101 "TN 11-09-11 1230 J. Sumner TVA Courier .0 7477 111109.22 TVA - Sequoyah Nuclear Plant - Intake 7N 11-09-11 1334 J. Sumner Murphys Courier 0.8 7509 111109.23 Duke Energy Corporation - McJuire NS - Outfall 001 NC SOP 34 - Exhibit G4.2, revision 06-29-09

BIOMONITORING CHAIN OF CUSTODY RECORD Page _1 - of -1 I' I nt: TVA Environmental Testing Solution, Inc. Delivered By (Circle One):

FedEx UPS Bus Client E e Ptject Name: Sequoyah NP Toxicity 351 Depot Street.

P4. Number: N/A Asheville, NC Other (specify):

General Comments:

Facility Sampled: Sequoyah NP 28801 NPDES Number: TN0026450 Phone: 828-350-9364 Rain Event? .

(Mark as Appropriate) ,L

___ ._., . ,Laboratorv Use Yes If Yes,_jNo Trace LETISLog _i P YyiLI

.4 3' k<II SQN-INT-TOX jComp ~'~;~ J1(2.5 gal) [iq[Jtl1tP Sample Custody - Fill In From Top Down '4 Sep&S i zr

~

_z~~r SO-f'.es tlec-ew'i es Relinquished By (Signature): Date/Time Received By (Signature): Date/Time -5A~tA1 i i o _ _ _ _ _ _ _ _

. O I g X/ _ _ _ _ _ _ E C_ _ _

_ _ _ ~t ~~~it t 1I__ ~ 1 -0 Instructions: Clients should fill in all areas except those in the "Laboratory Use" block. Biomonitoring samples are preserved by storing them at 6°C and shipping them in ice. The hold time for each sample is 36 hours4.166667e-4 days 0.01 hours 5.952381e-5 weeks 1.3698e-5 months from the time of collection. Therefore, please collect and ship in such a way that the laboratory will receive the samples with ample time to initiate testing within that time frame. Samples shipped overnight on Friday via FedEx or UPS must be marked for Saturday delivery or they will not arrive until the following Monday.

(/ý.

5* -QI Page _ S E I~

mm Whole Effluent Toxicity Sample Receipt Log r~ Date received Tho~m~

Tone received Received by Received from Sample temperature (*C)*

Project number Sample Sample name and description

-Sample temperature performed using Sample Receiving Thermometer: SN 6338.

State Comments 111111.01 Bladenboro WWTP NC 1- I1-II 0956 K. Keenan Fed - Ex I.1 7496 ian Fed - Ex 0.6 0.5 7497 7498 111111.03 111111.04 Progress W Energy -Carolinas allnasww-rP negyw-Darlas V*est erP SS.E.U ptemItk ae Fear

-- Cape taie NC NC I1-11-11I11III 0956 K. Keenan K. 111111.04 CiProfgGastuniae FearS._F._NC Fed- Ex 0.5 7498 I 1-11-11 0956 I Keenan 7499 111111.05 UnitedWater-EnfieldWWTP NC I1-1-I1 0956 K. Keenan Fed-Ex 0.7 in 7500 1 111111.06 Progress Ener*y Carolinas - Lee Steam Plant NC 1-1111 0956 K. Keenan Fed- Ex 0.5 As 7501 111111.07 Duke Enrgy Corporation - Marsh.all Steam Station NC 11 0956 ICKeenan Fed- E. 0.6 7502 111111.08 United Water - Scotland Neck WWTP NC I -I1I11 0956 K. Keenan Fed - Ex 0.7 7511 111111.09 ProgessEnery Carolinas-ShearonHarrisF-&E-Center NC I1-1111 0956 K_Keenan Fed-Ex 0.4 7490 111111.10 Microbac-Rockingh WWTP NC S1-11-11 1003 K. Keenan UPS 0.3 7491 111111.11 Microbac - Roseboro WWTP NC 11-11-11 1003 K. Keenan UPS 0.3 7492 111111.12 Microbac - J.P. Stevens - WagramWWTP NC I1-t1-11 1003 K. Keenan UPS 0.3 7508 111111.13 OWASA -Mason Farms WWTP NC Il-Il I1 1032 K. Keenan Dash Courier 1.0 7512 111111.14 Duke'oer Corporation - McGuire Nuclear Station - 1.9 Compusite NC _

1I-11 11 1235 J. Sumner M.rphy's Courier 27 TVA Courier 2.111.7 7477 ,TVAT-H111111.15 uoahNuerPlant-Oudll 101 I1-11-11 1220 J. Sumner 7477 111111.16 TVA - Sequoyah Nuclear Plant -Intake TN I1-11-11 1220 J. Sumner ITVA Courier, 2.0 SOP G4 - Exhibit 04.2, revision 06-29-09

Page 1 of 6 0ETS SEngionmentalTestingSolutl Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephalespromelas A.

Client: Tennessee Valley Authority County: ai4*w4 .3 Facility: Sequoyah Nuclear Plant 006 Outfall: 101 NPDES #: W44 0 *60TW o021'so *0 Project#: I f"t Dilutionpreparation information: Comments:

Dilution prep (%) 10.8 21.6 43.2 86.4 100 -

Effluent volume (mL) 270 540 1080 2160 2500 Diluent volume (mL) 2230 1960 1420 340 0 Total volume (mL) 2500 2500 2500 2500 2500 Test organism information: . Test information:

Organism age: IQ-.< 00 -.* OVN Randomizing template: 40-L.0, Date and times organisms AV-*-ii 116O Incubator number and were born between: shelf location: ,3L._

Organism source: Art'L 6,NLcM fA

-- *\ *" i Artemia CHM number: C-JiiSi pr Transfer bowl information:

Average transfer volume:

pH = 1.-0 Temperature =

2q.

S.U.

G°C Drying informationfor weight determination:

Date / Time in oven:

Initial oven emperature:

Date / Time out of oven:

Final oven temperature:

.t,.

O0 'C.

t o..

V-\

il2.A C)

Total drying time: "-L4geS Dailyfeeding and renewalinformation:

Day Date Morning feeding Afternoon feeding Test initiation, Sample numbers used MHSW renewal, or batch termination used Time Analst Time_ Analyst Time Analyst Outfall 101 Intake 0 Ivu-0*,1 ' hOG* ,h , tl0l~l ttea.. %.k,

-0+-IIA 2 'i-f'-lI-I1 WO'l1. .~.4. .J+/-...6 _k I'.o 01 ni alifl, 111-0q+lA 2 i,'=-II os .. . t .1.1..mJ1.01M-. IIW .)A I %%(.It IllJl\.\ ____-t 4 ) I\- l'-1 O'a, A II&SO .JL(III . IG It..

,,116. A-01-.%t 6

I~ %i('L-IN(., Control information: Acceptance criteria Summary of test endpoints:

% Mortality: (51. -520% 7-day LCso I>1007.

Average weight per initial larvae: 0.510 NOEC I 007.

Average weight per surviving larvae: 0.110 > 0.25mg/larvae LOEC >1C07.

ChV >=oo7.

IC2 5 T >1007.

SOP AT20 - Exhibit AT20.3, revision 04-01-09 Page 31 of 102 In 'ependt viewny Kelley E.Keenan Initials: !UP

Page 2 of 6 lETS I EnvimnmentalTesting Solutlon,, Inc.

Species: Pimephalespromelas Client: TVA / Sequovah Nuclear Plant, Outfall 101, Non-treated Date: Ii- os "I Survival and Growth Data Day CONTROL 10.8% 21.6%

A B C D E F G H I J K L 0 10 10 /* /0 ,Q /0 /0 /0- /0 /0 /0 /0 1 ' /3 /0 / 1/0 /0 '0 /0 /0 /6 /0 /0 2 (0 /0 /o /0 /0 /0 1 I /C ' o/0 I 3 10 10 10 (0 10 10 10 / '0 1 10 10 /0 I IoD 10o 0 1 o 1 10 M I0C) 10 to 5 10 10 0o /0 /0 /o 10 /0 1o t0 /0 /0 6_I0 /0 to /0 /O' "/C '0 /0 /0o t0 /0

__ __ __ to I~1 to lo If/0 to 1 0 A = Pan weight (mg)

Tray color code:: LI;jt\ 1p, .

Analyst: MM 'I.'i. d

%SA.. 14*4' l 461.L I- 11.t .(-4 130; 14.% 14.s(.

, ,q, Date: t1O.I Tr - I I Il B = Pan + Larvae weight (mg)

Analyst: M_

.. Z,. rl vA.i4 Date: 1.1q.1 l o.1% Ilq4 . ui V3

".. .1 .7 2.4.4,, 7.o.'AL 2. .5.4o Iq.o t..1%

C - Larvae weight (mg) = B - A bA q ( , 'r sl -'-

1.1 I -1.S1, .13 i.,q4 16-1, &M, 1.60 Hand calculated.

Analyst:

Weight per initial number of larvae (mg)

= C / Initial number of larvae Hand calculated.

Analyst:

0" ~0 ýý . & I'*ic iCy 42"I 01 ~0

~0 01

- I ~ I a i ___ - _____

Average weight per Percent reduction Initial number of from control() 01 0 0.1,41 1,:-7, 0-1LIO Al.7.

larvae (mig) I Comment codes: c = clear, d = dead, fg = fungus, k = killed, m = missing, sk = sick, sm = unusually small, Ig= unusually large, d&r = decanted and returned, w = wounded.

Comments:

i I

I Page 32 of 102 nrepenent review by Kelley E.Keenan SOP AT20 - Exhibit AT20.3, revision 04-01-09 Initials:

Page 3 of 6 IETS t Inc.

lutions, Species: Pimephalespromelas Client: TVA / Senuoyah Nuclear Plant, Outfall 101. Non-treated Date: .it04 -I

___Survival and Growth Data Day 43.2% 86.4% 100%

__M N 0 P Q R S T U V W X 0 1i I/0o 0 0 /0 10 t0 -0 30 ,0 t0o 0 10 /0 ,0 1O /0 '0 /0 ,010 /0 /0 /0 2 J0 /Q /0 /0 /0 'o 10 '00 /0 /0to 3 to to /C6 1'0 1C) IQC) /0 10 t /-6 /0 10 41L) Io) 10 10 /0 10 to 10 10 !t to 510 J 10 100 /0 ') 0 ( 0 '0/Q to 6 /D /0 /0 10 /0 'o /0 (0)1c/0 0 0 i 7__ _ _ o0t0 lb 10 ( ,I , to l lo '

A = Pan weight (mg)

Tray color code:: L.,-,v Analyst: _*.4* M'. (,t is.44 I 1. to4 14.1.,-.4 I t.4 lt.' l1.52 It1.". I -.4 14.l1 Date: \__"__

B = Pan + Larvae weight (mg)

Analyst: ________. 4______ ...

Date: it_._-1_

11.____ li . 1,o.o*$

U UA1.t, Z1L. ,. 7L.4 -A4

-4.5

'L.Ll e Z4.-

C = Larvae weight (nag) = B - A

.1 " 0 -, ll. . . QO A Al0 14 4,l Hand calculated.

Analyst:

Weight per Initial number of larvae (mg)

C / Initial number of larvae Hand calculated. IIP "' *" [* ()qO 4

I " QJ°o*

" ,.-.% ci O"6" J

-o g)

Analyst:

Average weight per Percent reduction initial number of from control(%) 0 -' "7 0.1( [ 1,.2.7.

larvae (mg)

Comment codes: c = clear, d = dead, fg = fungus, k killed, m missing, sk = sick, sm = unusually small, Ig = unusually large, d&r = decanted and returned, w = wounded.

Comments:

SOP AT20 - Exhibit AT20.3, revision 04-01-09 PaIe 33 of 102 Idpend t evewbyKelleyE. Keen Initials: X

Page 4 of 6 "ETS Envlmnmentallestlns SolutlonL Inc Species: Pimephales vromelas Client: TVA / Sequoyah Nuclear Plant. Outfall 101. Non-treated Date: it-w "I\

Day 100% Intake

_______________ Y Z AA BB 0 C 1L 0 /10 1

/0 10 2 /0 _ I0t

_________4__

(3 CIA to 1 5__ __ _ ' to t(

5/0 to ' 10""

7 144 A - Pan weight (rg)

Tray color code::

Analyst: mV A, 13Aj 14.04 IL.9 Date: _._._ _

B = Pan + Larvae weight (mg)

Analyst: MAC' Date: I_"_k.1_

C = Larvae weight (rng) = B - A Hand calculated. ( 1.6

. 1-011 Analyst: _ae _

Weight per initial number of larvae (mg)

= C / Initial number of larvae Hand calculated. /

Analyst: Wr Average weight per Percent reduction initial number of from control (%) O, 12. 1. e 7.

larvae (mg) I Comment codes: c = clear, d = dead, fg = fungus, k = killed, m = missing, sk = sick, sm = unusually small, Ig = unusually large, d&r = decanted and returned, w = wounded.

Comments:

SOP AT20 - Exhibit AT20.3, revision 04-01-09 I Pg

,nge34 8l penin revie~w~y Kelley E.Keenan Initials:

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated November 08-15, 2011 PimephalespromelasChronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 SETS Quality Control Verification of Data Entry, Calculations, and Statistical Analyses ni Environmental Testing Solutions, Inc. Project number. 7477 at Not for Compliance Assessment, Internal Laboratory OC Cae.tatira (V) Reptltate Inlitalmuamaerd Ramt oinaeahao A Pa.wti.ht(ani B-Paa+L-ao Lar-we.tght(naq) WtghtlSainmag Ma -lot Ightl Cotfliciatevariataaon Wthtbtllainail nber Means al Mao. eaight/2lltiat Coaiftd o Peeoat- .idard- f-lamrva w.elht(ram) -A-B ... berfhlrae(ata) Sor*lvo berof m-u l 11-a (aina(amg) (%) *umbotlertitae,. vaiLation *1_W coitrol (%)

aI~v.(.0 ***) % m A 10 10 13.12 20-11 6.99 0.699 0.699 Control B 10 10 12.63 19.44 6.81 0.681 0.770 12.0 0.681 100.0 0.770 12.0 Not applicable C 10 10 13.07 21.58 8.51 0.851 0.851 D 10 10 14.69 23.18 8.49 0.849 0.849 E 10 10 14.45 22.26 7.81 0.781 0.781 10.8% F 10 10 13.72 21.46 7.74 0.774 0.760 2.8 0.774 97.5 0.741 6.7 3.7 G 10 10 13.35 20.72 7.37 0.737 0.737 H 10 9 14.64 21.37 6.73 0.748 0.673 1 1 10 10 13.15 19.60 6.45 0.645 0.645 21.6% J 10 10 14.35 21.42 7.07 0.707 0740 12.2 0.707 3.9 K 10 10 14.56 23.15 8.59 0.859 0.859 L 10 10 13.64 21.14 7.50 0.750 0.750 M 10 9 13.59 20.79 7.20 0.800 0.720 43.2% N 10 10 14.62 22.19 7.57 0.757 0.723 10.9 0.757 97.5 0.703 8.6 8.8 0 10 10 13.44 19.60 6.16 0.616 0.616 P 10 10 12.86 20.03 7.17 0.717 0.717 Q 10 10 14.20 21.21 7.01 0.701 0.701 86.4% R 10 10 14.64 21.49 6.85 0.685 0.755 10.7 0.685 100.0 0.755 10.7 1.9 S 10 10 14.77 22.49 7.72 0.772 0.772 T 10 10 12.91 21.54 8.63 0.863 0.863 U 10 10 13.78 21.45 7.67 0.767 0.767 100% V 10 10 11.66 19.66 8.00 0.800 0.780 2.5 0.800 97.5 0.761 7.0 1.2 W 10 10 12.48 20.41 7.93 0.793 0.793 X 10 9 14.15 20.99 6.84 0.760 0.684 Y 10 10 12.91 20.04 7.13 0.713 0.713 100% Intake Z 10 9 13.93 20.88 6.95 0.772 0.751 6.8 0.695 95.0 0.712 2.2 7.6 AA 10 9 14.04 21.37 7.33 0.814 0.733 BB 10 10 12.85 19.91 7.06 0.706 1 0.706 Outfiall 101: MSD = Minimum Significant Difference Dunnet's MSD value: 0.1249 PMSD = Percent Minimum Significant Difference PMSD: 16.2 PMSD is a measure of test precision. The PMSD is the minimum percent differenrce between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Intake:

Dunnetl's MSD value: 0.0914 Lower PMSD bound determined by USEPA (10th percentile) - 12/0.

PMSD: 11.9 Upper PMSD bound determined by USEPA (90th percentile) = 30%.

Lower and upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET Intedaboratory Variability Study (USEPA, 2001a; USEPA, 2001b).

USEPA. 2001a, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes I and 2-Appendix EPA-821 -.- 01-004 and EPA-821-B-01-005. US Environmental Protection Agency, Cincinnati, OH.

File: sqn101_11O811data.xlsx Entered by: J Suwner Reviewed by: _

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated November 08-15, 2011 o ETS

,:1 Envlronen-UlT4U~ngSo1U1tnLM*&

Statistical Analyses Larval Fish Growth and Survival Test-7 Day Growth Start Date: 11/812011 Test ID: PpFRCR Sample ID: IVA/ SQN, Outfall 101 End Date: 11115/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Dlscharge Monitoring Report Sample Date: November 2011 Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelas Conc-% 1 2 3 4 D-Control 0.6990 0.6810 0.8510 0.8490 10.8 0.7810 0.7740 0.7370 0.6730 21.6 0.6450 0.7070 0.8590 0.7500 43.2 0.7200 0.7570 0.6160 0.7170 86.4 0.7010 0.6850 0.7720 0.8630 100 0.7670 0.8000 0.7930 0.6840 Intake 0.7130 0.6950 0.7330 0.7060 Transform: Untransformed 1-Tailed Isotonic Conc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Mean N-Mean D-Control 0.7700 1.0000 0.7700 0.6810 0.8510 12.035 4 0.7700 1.0000 10.8 0.7413 0.9627 0.7413 0.6730 0.7810 6.668 4 0.555 2.410 0.1249 0.7413 0.9627 21.6 0.7403 0.9614 0.7403 0.6450 0.8590 12.177 4 0.574 2.410 0.1249 0.7403 0.9614 43.2 0.7025 0.9123 0.7025 0.6160 0.7570 8.607 4 1.302 2.410 0.1249 0.7396 0.9605 86.4 0.7553 0.9808 0.7553 0.6850 0.1630 10.748 4 0.285 2.410 0.1249 0.7396 0.9605 100 0.7610 0.9883 0.7610 0.6840 0.8000 6.999 4 0.174 2.410 0.1249 0.7396 0.9605 Intake 0.7118 0.9244 0.7118 0.6950 0.7330 2.246 4 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test Indicates normal distribution (p > 0.01) 0.93824 0.884 0.0738 -1.0425 Bartlett's Test indicates equal variances (p = 0.86) 1.92626 15.0863 Hypothesis Test (1-taill, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnett's Test 100 >100 1 0.12491 0.16223 0.00226 0.00537 0.82787 5,18 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point % SD 95% CL(Exp) Skew IC05 >100 IC10 >100 IC1 - >100 1.0 IC20 >100 0.9 IIC25 >100o IC40 >100 0.8 IC50 >100 0.7.

o0.6 C

0C.5

o.41 0.3-0.2-0.1 -

0.0'--A 0 50 100 150 Dose %

Dose-Response Plot I

C, File: sqn10lIlOlldata.xlsx Entered by: J. Sueer PlnAe.tg14 i*94 Kelley E.Keenan Reviewed by: T Initials: _ __

TVA / Sequoyah Nuclear Plant, Outfall 101 - Intake Non-treated November 08-15, 2011 "ETS Environmental Testing Solutions, Inc.

Statistical Analyses Larval Fish Growth and Survival Test-7 Day Growth Start Date: 11/8/2011 Test ID: PpFRCR Sample ID: TVA / SQN, Intake End Date: 11/15/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring Report Sample Date: November 2011 Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelas Conc-% 1 2 3 4 D-Control 0.6990 0.6810 0.8510 0.8490 10.8 0.7810 0.7740 0.7370 0.6730 21.6 0.6450 0.7070 0.8590 0.7500 43.2 0.7200 0.7570 0.6160 0.7170 86.4 0.7010 0.6850 0.7720 0.8630 100 0.7670 0.8000 0.7930 0.6840 Intake 0.7130 0.6950 0.7330 0.7060 Transform: Untransformed 1-Tailed Conc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD D-Control 0.7700 1.0000 0.7700 0.6810 0.8510 12.035 4 10.8 0.7413 0.9627 0.7413 0.6730 0.7810 6.668 4.

21.6 0.7403 0.9614 0.7403 ' 0.6450 0.8590 12.177 4 43.2 0.7025 0.9123 0.7025 0.6160 0.7570 8.607 4 86.4 0.7553 0.9808 0.7553 0.6850 0.8630 10.748 4 100 0.7610 0.9883 0.7610 0.6840 0.8000 6.999 4 Intake 0.7118 0.9244 0.7118 0.6950 0.7330 2.246 4 1.239 1.943 0.0914 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.93334 0.749 -0.0275 -0.8321 F-Test indicates equal variances (p = 0.02) 33.6016 47.4683 Hypothesis Test (1-tail, 0.05) MSDu MSDp MSB MSE F-Prob df Homoscedastic t Test indicates no significant differences 0.09137 0.11866 0.00679 0.00442 0.26167 1,6 Treatments vs D-Control Dose-Response Plot 1

0.9 0.8 0.7 .... ...... i1-tail, 0.05 level of significance 0.6' C: 0.5 0 0.4 0.3 0.2 0.1 0

o C~40 o3 coS C COC File: sqnl0l_110811data.xlsx PAnnfnt9efvl,'y Kelley E.Keenan Initials:V' Reviewed by: +/-

Entered by: J. Sumner

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated November 08-15, 2011 Pimephalespromelas Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 SET'S Daily Chemical Analyses Project number: 7477 F Environmental Testing Solutions, Inc.

Da 0 Da I Da 2 Da 3 Da 4 Da 5 Da 6 Concentration Parameter Initial IFinal IInitial IFinal Initial IFinal iInitial IFinal IInitial I Final IInitial IFinal IInitial IFinal Control Alkalinity (mg/L CaCO 3)

Hardess (mg/L CaCO3) 24.8 24.7 24.9 24.3 24.8 24.7 24.9 24.7 24.8 24.6 24.9 24.7 24.7 24.8 Temperature (C 7.87 7.88 8.05 7.73 7.87 7.72 8.07 7.861 7.96 7.66 8.06 7.83 8.031 7.74 PH (SU 7.9 7.7 7.7 7.7 7.8 7.0 7.7 7.6 7.6 6.8 7.7 7.6 7.6 7.5 DO m 303 300 300 296 293 295 293 10.8% Conductivi hos/cm 24.9 24.5 25.0 24.7 24.8 24.9 25.0 24.3 24.9 24.8 25.0 24.5 24.7 24.6 Tem erature C) 7.87 7.86 8.06 7.73 7.89 7.69 8.10 7.86 7.97 7.68 8.041 7.80 8.04 7.78 PH (SU 7.8 7.7 7.7 7.6 7.8 7.0 7.7 7.6 7.6 6.9 7.6 7.5 7.6 7.4 DO m 21.6% Conductivi hos/cm 286 286 283 284 283 2 281 24.5 25.0 24.6 24.9 24.8 25.0 24.6 24.9 24.7 24.9 24.5 24.7 24.5 Tmeaue24.9 2.1 2 2.0 2.8 2.1 2.7 2.9 2.5 2.9 24 2.8 2.5 Tern eratnre C 2.9 2.6 43.2% Conductivi hos/cm) 24.8 25.0 24.7 25.1 24.5 24.9 24. 24.9 24.6 24.8 24.7 Tern erature C) 25.1 24.8 25.1 0 0 0 ý 0 1 0 8.% Conductivit (imhos/em) 246 2. 245 2.1 2.1 2. 247 2. 245 2.1 46 2.845 Temperature OC)2.

s1018 ý191 16 Conductivit (phos/cm) 100% Alkalinity (mgfL CaCO 3)

Hardness (mg/L CaCO3) 77 Total Residual Chlorine (m.L)

24. 24.6 25.0 24.7 25.0 24.7 24.9 24.5 24.9 24.5 24.9 24.6 24.8 24.6 Tern erature C) 100% Intake Alkalinity (mg/L CaCO3)

Hardness (mg/]L CaCO3)

Total Residual Chlorine (mg/L) 47 49 45 2.1 2.1 2.1 2.1 2.1 2.

-Temperature *CC)2.1 2.1 2.1 2. 50 File: sqn101_110811chem.xls Entered by: C. Johnson Reviewed by: 4

Page 5 of 6 "ETS I

I EnvirnmentalTesting Solutions, Inc.

Species: Pimephalespromelas Date: ll-Le* \

I Client: TVA / Sequoyah Nuclear Plant. Outfall 101, Non-treated Daily Chemistry:

I Dav I I Concentration IParameter Analyst p CONTROL Non-treated Conductivity himhos/ern)

Alkalinity 4

(mgnCaCOSU) a Hardness (mc CaCOd/L)

Temperature (0C0 a - _ _ ý pH (S.U.)

DO (m-/L)

300o 300 10.8% Conductivity (Aimhos/cm)

I 00oo Temperature (0C0 4 -, I PH (S.U.)

t ~

IS-, . 1qo DO (mg/L) 21.6% Conductivity (amhos/cm)

Temperature (°C) 43.2% Conductivity 1S.0 imhos/cm)

____________ TemMerature (0C) L*- +.o' .la L.Q L.

nH (S.U)' .4.M 000 DO (rag/L) 86.4% Conductivity Temperature (°CQ 111.1.9 2S. k U-0 7 -pH (S.U.) *q 4-S t¢.

Conductivity ne Alkalinity 100~~_(mg CaCO 3 "L)

Hardness T- Tl (mg CaCO 3/L) 1 TR chlorine (mg/L)

Temperature (°C) 2S. ,

nH (S.UI .1 LJ..-LL---41--...I.-.,

DO (mr/.,i R.ro Conductivity (gmilos/cm) -_____

100% Alkalinity Intake (Mg CaC0 3 1L) ____

Hardness (Mg CaCo 3/L) _______

TR chlorine (mg/L) 40.10 Temnerature (ofl I 2(4.

I.- . 1i 0.. I " - - -" "

Initial II Final I Initial 11 Final I Initial 11 Final I SOP AT20 - Exhibit AT20.3, revision 04-01-09 P"aIfid*pL'tn'**v1iy Kelley E. Keenan Initials:

Page 6 of 6 IG i

oETS EnvironmentalTesting Solutions, Inc.

Species: Pimephalespromelas Client: TVA / Seaunuvah Nuclear Plant. Outfall 101. Non-treated Date: it I I Day I 3 I Analyst F 11 r~ir A IA Pe-r" Concen- Parameter tration I pH (S.U.)

DO r(rag/L)

Conductivity 34-0 (umhos/cm)

CONTROL Alkalinity Non-treated (me CaCOJL) 4.1 Hardness (me CaCO,/L)

I. bznvý II -6, I *q---"-I "A Temperature (0C) II ,,A,

- a a - _______________________________________________

nH (S.U.) II 3-4C. I ~ II 0.

Isao 1 pH (S.U...)

r m DO (mg/L) 4A'#II 10.8% Conductivity rI (gmhos/cm) ____

W-LA6 i-+¶~o Temperature (0C) M .0 II S -LI.6 a a~

U 0.04 pH (S.U.)

21.6%

DO (mg/L)

Conductivity Qirnhos/cm)

[ a . [(, I ,

elTemnerature (°C* (Q L¶.%I I a Temperature DH (S.U.)

DO (mi/L) 4.iO 910 I ^%.LA II _ Ato 00 ~i 43.2% Conductivity (umhos/cm) IS I-I0 "LAp O.f Temperature (MC)

- *1 a - 4 nH (S.U.)

(S.U.)

.t II % '.

OH DO (m&/L) Ua3TO 86.4% Conductivity

(,jmhos/em)

Temperature (0C0

=A0(o S1_i J I - *I~

"t., \ II IQ " a.j 1q.3. *,9 Wo"

'20 A pH (S.U.)

w0A~- rII DO (mg/L) I "-9~

Conductivity (tgmhos/cm)

Alkalinity 100% (mg CaCO 3IL)

Hardness (mg CaCO 3/L) 2,1-195 TR Chlorine (mg/L)

Temperature ('C) ij* t p -

DH (S.U.)

I n ii " " I --

DO (mgiLn

-. z-...-

Conductivity 100%

mgrhosfcmn Alkalinity 7Q-S ,

Intake nmCaCO 3/L)

Hardness (mg CaCO 3/L)

TR chlorine (mg/L)

Temperature (°C) a I I I Initial II Final I Initial II Final I Initial U Final I Initial I1 SOP AT20 - Exhibit AT20.3, revision 04-01-09 Page 40 of 10 2? Kelley E.Keenan Inotpent~nt reviewoyely .Kna inital..

Page I of 7

-*ET Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1002.0)

Species: Ceriodaphniadubia Client: Tennessee Vialley Authority County: Hamilton Facility: Sequoyah Nuclear Plant Outfall: 101 NPDES #: TN0026450 Project #: NL131 Dilutionpreparationinformation: Comments:

Dilution prep (%) 10.8 21.6 43.2 .1 86.4 100 Effluent volume (mL) 270. 540 1080 2160 2500 Diluent volume (mL) 2230 " 1960 "1420 340 0 Total volume (mL) 2500 2500 2500 2500 2500 Test organismsource information: Test information:

Organism age: I < 24-hours old Randomizing template color.

Date and times organisms were born It614-l1 I OWL 0ncubator number and shelf Culture between:board: I A- 011 A.i Inlocation: oato:2 n

Replicate number: 1 2 I3 4 15 7 1 9 1-9 1 10 YTbth Culture board cup number. II'. I O1 " ". Z,32.

I

1-1153I , YWT batch:

Transfer vessel information: IH = J.1S S.U. Temperature="LS00C Selenastrum batch:--

Average transfer volume (mL): I . Se ubcI I Daily renewal information:

Day Date Test initiation and feeding, MHSW Sample numbers used Analyst renewal and feeding, or batch used Outfall 101 Intake termination time 0 11- s-il1 _ _ I-04 H__*- A if 11101,01 it ...- A 1 1o~i-ll 69SQ iiOcA-11A 1111 (n.at %X41% T1. ...j:....

2d 114o1 LNl*t-N*-AIt I>f\t 0A. It 1\ tt Os. - A*

3 II'A-S

L. ti-og-II & , kw as. -4 . .\" I0I . - ,,*L 4 11-12.11 1O__

a1-_ _._n ___

I_ __L_ 111111.16"

______ ____, __ _ _.- _ i - I& iIt l ,tS t l.16h 6

11-M -11 I11 mt. IS 111 1-. IA.

7 .1000 Control information: Summary of test endpoints:

_ Control-I Control-2 Acceptance criteria

% of Male Adults: 0*) 0___7. <_20% 7-day LC50 >_ IW1,

% Adults having 3 rd Broods: 1007..

10:7* > 80% NOEC I007.

" Mortality: 0,()I, -! 20% LOEC 5 1067.

Mean Offspring/Female:

%CV:

?. I 5-0. ".37.

21..(a 15.0 offspring/female

< 40.0%

ChV IC25 j 1*ii7.

>_too'.

Pl#AVed#k)fe1 by Kelley E. Keena n SOP AT 11- Exhibit AT 11.2, revision 06-01-11 Initials:

Page 2 of 7 lETS Species: Cerlodaphniadubia Client: TVA / Seqluoyah Nuclear Plant. Outfall 101 Date: Wi-1A CONTROL-1 Survival andReproduction Data Replicate number Day _ 1 2 31 4 5 6 7 8 9 10 1 Young produced ' cŽ .* '

1 Adult mortality I . L -. ___ _-- _

21 Young produced L~~.0 0 0 01 I20 0

Adult mortality .- 1 '1 '- 1° 3 Adult mortality Young produced 0 ) *C i L_ 1 \. I %.- T3-Iac %)I*.-

4 Young produced 'S '[%

Adultmortality 11],_i0__

6 5 oung produced Young produced I*1**I**]**

Adult mortality D 1 16iQ1.S2%0 I*I*";***I" 0

I C)0 _

1 C) 0 C%*

0 I' C:)1 Total young produced W It - V-%t 5.go0 ? s o*

ordBos.

Final Adult Mortality _C7 * *... \. s_

Xfor 3' Broods 5c-. *x. t- *-7 5Z7.-"*

Note: Adult mortality (L - ive, D dead), SB = split brood (tingle brood split between two days), CO = carry over (offspring carried over with adult during transfer).

Concentration:

%Mortality:____

Mean OffspringFemale:

CONC: 10.8% Survival andReproduction Data Replicate number Day 1 2 3 4 15 6 7 8 9 10 1 Young produced () , 0 0 0 C 0

Adult mortality LJ _ %..- %.-

_ Adult mortality C i'-1' C-' I L_- V__I 4 Young produced Adult mortality - - j1 l +/- __ _

AdultYoung produced mortality 1(0l[_*

%. 17 1, U tIl _*L..l. (31 C7*.

dut_

_ _ Young _L produce 0 0 I 0 - 0_- (D Autmortality - -_ -I -I'- L. L Total youngproduced ! - . , * .. .- I,..,..,_

Final Adult Mortality - ?

t - - L_ I.-.LL_

Note: Adult monality (L = live, D = dead), SB - split brood (staile brood aplit between two days), --cOan over (offspringcarnedover with adult during transfer).

Concentration:

% Mortality: 0W.

Mean Offspring/Female: SO.1

% Reduction from Control-1: - .f.q 7.

SOP ATI 1 - Exhibit ATI 1.2, revision 06-01-11

.1Initials:

Ir-nQtqa___ "i_ . " E. Keenan Kelley

Page 3 of 7 JETS Species: Ceriodadhniadubla Client: TVA / Sequoyah Nuclear Plant, Outfall 101 Date: _ _

_-_ _i_

CONC: 21.6% Surviival and ReproductionData Replicate number Day 1 2 3 4 5 16 17" 8 9 10 I Young produced o (t3 n) o* 6) *0* C) 0 0 Adult mortality _ _V. t 2 Young produced ) I I]1 _

... [ Adult mortality t.

Youngproduced L i. .] Q, _ . ,. ._ .

2

, Adult mortality 4 Young produced t I C L-l.. 1 5_Young produced Adult mortality il Zf 1YIIVp I 2 Z Adult mortality

[ 't.LiA I 75- Young produced 61 Young produced IdI"ti. C"001 ]0IOD[Ii I~I aI 1 l 0Q I *T I

_ Adult mortalty j J ~ j'.. -I' cI'I~ Is, TYoung produced t- .16 I'll Ill Is i Total young produced * *o .-- 3 5 31 Final Adult Mortality . . ,~. I - ._ .. ** *...

Note: Adult mortality (L - live, D dead), SB - split brood (single brood split between two days). CO ca over (ofsedng carried over with adult dunng tansfer).

Concentration:

%Mortality: (7 Mean Offspring/Female: ,1 .

% Reduction from Control-1: - 1*

I CONC: 43.2% Survival andReproductionData Replicate number Day 1 2 3 1 4 1 5 6 7 8 9 10 1 Young produced I 0_

(J 1 0' j( 5 0 I Adult mortality L ...

I.-- L , L_ %__

2 Young produced -' ) 0C) _

Adulmortality _,- %.-_

I 3 Y Youngproduced C, G 0 1 In Adult mortality 4 Young produced 4 (11 1 WL S__

I Adult mortality L . [ ... L. L-J U.

5 Young produced It 2JJJ1 I I~I.I..

6 Adult mortality

Yung produced Adult mortality 01_

%--~

M J \-j\-'- -

Li K =-t--

=1 I 7 Young produced Total young produced II 34 20 IS

" Z1 i

.0 I"

3 3

I*

.3S

" I *t 5-6 3S

'Final Adult Mortality %t !.- L. - ]a L. LL.

Note: AdUltmortality (L = eive,D = dea), StB= split broad (single brood split between two days), Cu = ca ovseprJtspnnrg canm over with adult durng transfer).

Concentration:

%Mortality: G ".

Mean Offspring/Female: Ab.I

% Reduction from Control-I: -A.17a Pa In 'penu*,K43 of

_e 10 Kelley reviewoy e E. Keenan SOP AT 11- Exhibit ATI 1.2, revision 06-01-11

. Iýnitials:

Page 4 of 7 SET

ndmnnmlmdTe.*fla SrUnt,.

Species: Ceriodaphniadubia Client: TVA / Sequoyah Nuclear Plant, Outfall 101 Date: It-04F11 CONC: 86.4% Suryiival and Reproduction Data D CpnInt , t

[L nulmber IlU*Ubl rs~v 1 2 3 4 N 6 7 S 9 10 Da Young produced 14 2 3-Q 10 0 1 0 L*) 0 _

Adult mortality _ \ 1 - I '.. -_. - '

2 Young produced 1C1)f 0 0 0 SAdultmortality

____ltI_._EKI l *

\.-

L...

3 Young produced I p Op OT I_0 W I 0I

_ Adult mortality I 4_i..I LI -I I _ - I-I -_

4 _

Y oung produced It16.lI-Adultmort*itylI '

I l t __

- lt

__ 1 * -

T l1+-I I l iL--I[ L 5 Young produed Ic .l Il .I A '2. 1 10 2. lO I Adult mortality _ '-I _'I - .- I --- "-V---

6 7

Adult mortality Young produced

__~

14l 16

- K]

La -VI za 5I .

I _

Total young produced 810 s.AJ ,q * * *' *,

Final Adult Mortality I I...V-I . I - - . -

Note: Adult mortality (L = five, D dead). SB - split brood (single brood split between two days), CO - can, over (oftprini carted over with adult during transfer).

Concentration:

% Mortality: . 7.

Mean Offspring/Female: I *O

% Reduction from Control-I: "14.4'?.

CONC: 100% Survival andReproduction Data Replicate number Day 1 2 3 14 5 16 71 8 9 10 I Young produced n C)

Adult mortality L LaL4L- uV.*IL [_Lc L I) 2 1on produced '1 1..QI~I Adult mortality r __ L_

I A L l*._ i 6 _ 0 ... ,*.

3 _Young produced _ In Adult mortality L -. L V. _I 4I Young produced Adult mortality LL IIL 5 Young producedI'1j 0 I..[L 1012 2. I IIl'L fAdult mortalit~y J ~ L -'- -1L '-l-~ I 6 Youngproduced I )I I C) O IO[ 0 C) I© I_. Adult mortality I~=%,- IW'J_-

L~-[- %I 'i-7 Young produced T ota l y o u n g p r o d u c ed j 4 L-3 0% .36

, t 3

I I 1' -

10 Qi 15

. C3___

"2 S-S 3 -6*s3 q Final Adult Mortality . . -. * * . I '. I ' . .

Note: Adult mortality (L - live. D - dead). SB - split brood (single brood split between two days). Li) catt = over ontpnng careo over w am tult dunag ýansaerJ.

Concentration:

% Mortality: 07.

Mean Offspring/Female: 3&

% Reduction from Control- 1: -7.4 ]

SOP ATI I - Exhibit AT 11.2, revision 06-01-11

Kelley E. Keenan

! :ETS Page 5 of 7 Species: Ceriodaphnladubia Client: TVA / Sequoyah Nuclear Plant, Outfall 101 Date: Itok"It.

CONTROL-2 Surviival andReproductionData Replicate number Day 1 2 3 4 5 6 7 8 9 10 2 Young produced C Adult mortality ~_ _ I.L..

L 2 Young produced c0 0 0 0 1 0

_J _ Adult mortlity I L-Adult mortality _____ I ____ LI[Ll %.-Il~

L**** I__ I'-_ _ I___ I I__ L I I Young produced =o 31

_ Adultmortality 5

6 6 I

[ Young produced Adult mortality Young Yunpoue produced *'-

Adult mortality ,, 1%

I i-

[

-J ---_

t 94.. J 0. -

'~_'-" I kII I

1,5 %-6 b

I '-

'7 [Young produced t__ ,t j t* t I t¢ I * " ** I Total young produced 711 11 7 n .- - 3 I 2* . 2.7 Final Adult Mortality X for 3r Broods

Ž1. ']+/-1 .

Note: Adult mortality (L - live, D - dead), SB split brood (single brood split between two days), CO = carry over (offspring canTiedover with adult during transfer).

Concentration:

CONC: 100% Intake Survival and ReproductionData

%Mortalit:w Mean Offsprinlemnale: 21.I " I Replicate number Day 1 2 3 4 5 6 7 8 9 10 1 Young produced 0n C)

Adult mortality L ) L JLJ%- 1-L( L'( L(

.2 Young produced C 013 1~0 10 I0 I0 I Adult mortality ___ I %-- L_ k LI I L",

3 Young produced Adult mortality l _ ILl

0. ls LI L'-

0 1*CO Young produced 1 % 1 IJ.L 1 7F q Adult mortality _ T I QI I_ I I 5 Young produced -

Adult mortality _ L I-- I L IL I 7 ]Young produced l'* Zo \%C ttA lo'I I Total young produced T -

I ' j - I 3

I 2' Final Adult Mortality k : k-.-- ..- A.- -

L..

?

I -.

,. -A-I U

.. t - o Iy k.10 - " ...-ý O .. M" . - .I.-- - - -,a oc - 111 Concentration:

% Mortality: 01-I erfla Initials:

'! La l'94Kelley E.Keenan Mean Offspring/Female:

% Reduction from Control-2:

SOP ATI 1 - Exhibit AT 11.2, revision 06-01-11 33 .. L

- o. 70.

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated November 08-15,2011 00ETS Verification of Ceriodaphnia Reproduction Totals

) E ontralTesthVSol-t IS loerV CD~ Control-I 86.4%

Replicate number Total CD Day number

'licate Total Day 1 2 30 4 6 7 1 8 1 9 1 10 8 9 1 0

-X(S 0 0 10 101 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 3 0 0 -0 0 0 -0 0 0 0 0 0 1t 0 flIAI l lf0 tt 0O0OI 0 4 5 6 5 4 4 4 5 4 6 5 48 4 5 3 3 3 4 3 4 5 3 3 36 5 12 13 11 11 12 0 1 11 13 13 118 5 9 11 11 10 1I 10 10 9 9 10 100 6 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0 0 16 0 17 0 13 146 0 7 19 16 18 19 18 20 20 17 19 18 184 7 16 13 15 15 14 15 12 28 26 30 29 26 282 Total 36 35 34 34 34 34 37 32 3L36 350 Toa 0 2L7 2 29 10.8% 100%

DyReplicate number Total Re plicate number Total D 1Y 1 2 3 4 5 6 7 8 9 10 Day 5 6 7 8 9 10 21 2 3 4 1 0- 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0- 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 4 4 4 4 41 4 5 4 5 5 6 5 5 4 4 4 47 4 4 5 4 3 4 5 12 11 12 5 ll. 10 11 10 12 11 11 13 10 10 109 $ 13 12 13 12 10 12 11 118 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 l,7 17 N17 15 1 14 19 116 15 19 17 167 7 16 19 21 19 18 19 18 20 18 18 186 23 31 3 2 33 31 317 Total 34 35 39 36 34 36 35 35 33 34 351 Total 32 321 30 32 21.6% Control-2 Dy IRe* ticate number 8 0 Total DyRep)licate number Total DY 1 213 4 5 6 7 8 9 10 Day 1 2 3 4 5 6 7 8 9 10 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 00 00 00 00 0 0 3H 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 4 5 4 4 4 4 5 4 4 J 4 q41 4 3 3 4 4 4 3 3 3 3 3 33 5 12 10 12 11 10 10 12 10 13 10 110 5 10 10 9 10 12 10 10 10 10 11 102 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 0 15 19 195 15 164 7 14 14 14 13 14 16 15 13 14 14 141 7 18 20 15 20 18 16 302 32 31 35 33 31 29 315 Total 27 27 27 27 30 29 28 26 27 28 276 Total 34 31 43.2% 100% Intake DyRep~icate number 9 0 Total DyReplicate number Total 2 1 3 4 5 1 6 7 8 9 1 Day 1 2 3 4 5 6 7 8 9 10 Da 1 I 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 "0 0 4 4 6 4 4 4 5 5 4 5 5 46 4 4 3 4 5 5 5 3 5 4 4 42 5 12 12 10 10 10 11 10 12 12 12 111 5 11 13 13 12 12 11 12 11 11 11 117 6 0 0 -0 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0 7 18 20 15 20 18" 1 7 1 '5 19 19 18 179 7 o 17 20 15 1 '15 17 17 15 19 19 173 Total 34 r8- 2-9 34 r2 r3 SU 3.1 _21 35 36 1 Total 132 136 132 361 32 133 32 31 34 34 332 File: sqn101_110811data.xlsx Entered by: J. Surrper Reviewed by:

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated November 08-15, 2011 Ceriodaphniadubia Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1002.0 E S Verification of Data Entry, Quality Control Calculations, and Statistical Analyses (D Environmental Testing Solutions, Inc.

on Project number: 7477 GJ Concentration Replicate number Survival Average reproduction Coefficient of Percent reduction front

(%) (offspring/female) variation (%) control (%)

4 S 6 7 8 9 10 28 29 28 26 30 29 26 100 28.2 5.2 Not applicable 31 30 35 31 32 33 31 100 31.7 4.7 -12.4 32 29 31 35 33 31 29 100 31.5 6.4 -11.7 34 32 33 30 35 36 35 100 33.6 8.1 -19.1 34 34 34 37 32 38 36 100 35.0 5.0 -24.1 36 34 36 35 35: 33 34 100 35.1 4.7 -24.5 27 30 29 28 26 27 28 100 27.6 4.3 Not applicable 36 32 33 32 31 34 34 100 33.2 5.3 -20.3 Outfall 101: MSD = Minimum Significant Difference Dunnett's MSD value: 1.946 PMSD = Percent Minimum Significant Difference PMSD: 6.9 PMSD is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Intake:

Dunnett's MSD value: 1.156 Lower PMSD bound determined by USEPA (10"' percentile) = 13%.

PMSD: 4.2 Upper PMSD bound determined by USEPA (90d' percentile) = 47%.

Lower and upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET Interlaboratory Variability Study (USEPA, 200 1a; USEPA, 2001 b).

USEPA. 2000. Understanding and Accounting for Method Variability in Whole Effluent Toxicity Applications Under the National Pollutant Discharge Elimination Program. EPA-833-R-00-003. US Environmental Protection Agency, Cincinnati, OH.

USEPA. 2001a, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes 1 and 2-Appendix. EPA-821-B-01-004 and EPA-821-B-01-005.

US Environmental Protection Agency, Cincinnati, OH.

File: sqn101_110811data.xlsx Table populated from associated "Verification of Ceriodaphnia Reproduction Totals" spreadsheet.

Spreadsheet entered by: J. SumrWr Reviewed by:

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated November 08-15, 2011 0rFTn Statistical Analyses Certodaphnia Survival and Reproduction Test-Reproduction Start Date: 11/8/2011 Test ID: CdFRCR Sample ID: TVA / SQN, Outfall 101 End Date: 11/15/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring Report Sample Date: November 2011 Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Cerlodaphnia dubla Conc-% 1 2 3 4 5 6 7 8 9 10 Control-1 30.000 27.000 29.000 28.000 29.000 28.000 26.000 30.000 29.000 26.000 Control-2 27.000 27.000 27.000 27.000 30.000 29.000 28.000 26.000 27.000 28.000 10.8 32.000 32.000 30.000 31.000 30.000 35.000 31.000 32.000 33.000 31.000 21.6 34.000 31.000 30.000 32.000 29.000 31.000 35.000 33.000 31.000 29.000 43.2 34.000 38.000 29.000 34.000 '32.000 33.000 30.000 35.000 36.000 35.000 86.4 36.000 35.000 34.000 34.000 34.000 34.000 37.000 32.000 38.000 36.000 100 34.000 35.000 39.000 36.000 34.000 36.000 35.000 35.000 33.000 34.000 Intake 32.000 36.000 32.000 36.000 32.000 33.000 32.000 31.000 34.000 34.000 Transform: Untransformed 1-Tailed Isotonic Conc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Mean N-Mean Control-i 28.200 1.0217 28.200 26.000 30.000 5.233 10 32.517 1.0000 Control-2 27.600 1.0000 27.600 26.000 30.000 4.253 10 10.8 31.700 1.1486 31.700 30.000 35.000 4.714 10 -4.113 2.287 1.946 32.517 1.0000 21.6 31.500 1.1413 31.500 29.000 35.000 6.393 10 -3.878 2.287 1.946 32.517 1.0000 43.2 33.600 1.2174 33.600 29.000 38.000 8.084 10 -6.346 2.287 1.946 32.517 1.0000 86.4 35.000 1.2681 35.000 32.000 38.000 5.040 10 -7.991 2.287 1.946 32.517 1.0000 100 35.100 1.2717 35.100 33.000 39.000 4.739 10 -8.109 2.287 1.946 32.517 1.0000 Intake 33.200 1.2029 33.200 31.000 36.000 5.275 10 Auxiliary Tests Statistic Critical Skew Kurt Kolmogorov D Test Indicates normal distribution (p > 0.01) 0.45676 1.035 0.15802 0.20153 Bartlett's Test indicates equal variances (p = 0.41) 5.05964 15.0863 The control means are not significantly different (p = 0.33) 1.00623 2.10092 Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnett's Test 100 >100 >100 1 1.94578 0.069 68.6967 3.62037 7.6E-11 5,54 Treatments vs Control-1 Linear Interpolation (200 Resamples)

Point % SD 95% CL Skew IC05 >100 IC10 >100 IC15 >100 1.0 IC20 >100 0.9 lIC25 >100 0.8 IC40 >100 0.7 ICS0 >100 0.6 0.5

0.4 L 0.3 0.2 0.1 0.0. 0 4 0

-0.1 '.. .

-0.2 . . . .

-0.3

-0.4 . . . . . . . . ,,,.

0 50 100 150 Dose %

Dose-Response Plot 45 40 30 1-tail, 0.05 level 225 of significance 13

15 10 5

8 8 File: g91t01I1081Idataxlsx lK y E. Keenan by: J. Suker REntered Initials: ' .,ee Rýeedb

TVA / Sequoyah Nuclear Plant, Outfall 101 - Intake Non-treated November 08-15, 2011 S

ETS nmeM&ThAvbo ing I Statistical Analyses Ceriodaphnia Survival and Reproduction Test-Reproduction Start Date: 11/8/2011 Test ID: CdFRCR Sample ID: TVA / SON, Intake End Date: 11/15/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring Report Sample Date: November 2011 Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Ceriodaphnia dubia Conc-% 1 2 3 4 5 6 7 8 9 10 Control-1 30.000 27.000 29.000 28.000 29.000 28.000 26.000 30.000 29.000 26.000 Control-2 27.000 27.000 27.000 27.000 30.000 29.000 28.000 26.000 27.000 28.000 10.8 32.000 32.000 30.000 31.000 30.000 35.000 31.000 32.000 33.000 31.000 21.6 34.000 31.000 30.000 32.000 29.000 31.000 35.000 33.000 31.000 29.000 43.2 34.000 38.000 29.000 34.000 32.000 33.000 30.000 35.000 36.000 35.000 86.4 36.000 35.000 34.000 34.000 34.000 34.000 37.000 32.000 38.000 36.000 100 34.000 35.000 39.000 36.000 34.000 36.000 35.000 35.000 33.000 34.000 Intake 32.000 36.000 32.000 36.000 32.000 33.000 32.000 31.000 34.000 34.000 Transform: Untransformed 1-Tailed Conc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Control-i 28.200 1.0217 28.200 26.000 30.000 5.233 10 Control-2 27.600 1.0000 27.600 26.000 30.000 4.253 10 10.8 31.700 1.1486 31.700 30.000 35.000 4.714 10 21.6 31.500 1.1413 31.500 29.000 35.000 6.393 10 43.2 33.600 1.2174 33.600 29.000 38.000 8.084 10 86.4 35.000 1.2681 35.000 32.000 38.000 5.040 10 100 35.100 1.2717 35.100 33.000 39.000 4.739 10 Intake 33.200 1.2029 33.200 31.000 36.000 5.275 10 -8.400 1.734 1.156 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.91162 0.868 0.74434 -0.2836 F-Test indicates equal variances (p = 0.25) 2.22581 6.54109 The control means are not significantly different (p = 0.33) 1.00623 2.10092 Hypothesis Test (1-tail, 0.05) MSDu MSDp MSB MSE F-Prob df Homoscedastic t Test Indicates no significant differences 1.15604 0.04189 156.8 2.22222 1.2E-07 1,18 Treatments vs Control-2 Dose-Response Plot 40 .

40 35.

I 30 25

.I..................

1,.1111111ýý 1-tail, 0.05 level of significance

&20 15 10 5.

u . ..

GO C (C4 Nt 0 0C 8 8 " m 0 C~-

File: sqnlOl_110Blldata.xlsx Entered by: J. Su er Plr@Ppn 1+95 119; Kelley E. Keenan Reviewed by:

Initials: _

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated November 08-15, 2011 Ceriodaphniadubia Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1002.0 jtOT aX "Daily Chemical Analyses Environmental Testing Solutions, Project number:

Inc.

F5 - I -

Concentration IParameter Day0 I Day I Day 2 (m*L) Initial I Final I Initial I Final I Initial I a PH (IS 8.021 8.05 - 7.871 7.90 DO (mp/L) 80 77 Conductivity (pmhos/cm) L Control Alkalinity (mg/L CaCO3) 87 Hardness (mg/L CaCOM) 4 -

.... ~I ~

_______ emperature IL) hos/cm 24 24.9 2,5 20 4 4,82,,24. Z4.51 24.9 z5.:z 2 24 249 24.81 25.11 24.91 25.01 L C hos/cm PH DOm hos/cm 7.87 802 .05 7.871 7.87 80 8.071 7.971 7.96 803.06 80 8.03 79 DO (mg/L)

Conductlvi 10.8% hos/cm 7.9 79.779 7 7. 6 7.7 7.6 76.8775 7.67.

erature hos/cm 10.8% Conductivit (pho/em) CaCO3) hos/cm 303 300 300 296 293 295 293 rem CaCO3)

_________Temperature m CC)CaCO3) 4 24.9 252.20 24.8 29 24.8 24.82. 249 50 249 24. 8 25.0 2.

pH (SU CaCO3)

DO L 7.87 8.0 806 7.88 7.89 80 8.10 79 7.97 8.02 8.04 8.02 8.04 7.92 Conductivi 21.6%

DO (mn!L) 7.8 7.9 7.7 7.9 7.8 7.6 7.7 7.6 769 7.6 7.6 7.6 7.7 21.6% Conductivity

°C (junhos/em) 286 28,6 28(3 284 281 C

Chlorine 278 281

_______Temnperatur CC) 24.9 .25.1 25.0 24.8 24.9 252 24.8 25.2 24.9 20 24.9 24.9 25.0 24.9 PH(SUI)

DOm (mg/L 7.88 8.02 8.06 7.89 7.88 8.0 8.10 800 7.97 85 7.96 804.05 79 Conductivi 43.2%

DO (n91L) (mg/L 7.8 7.9 7.8 7.9 7.8 7.6 7.8 7.7 7.7 79.6 7.8 7.6 7.7 43.2% Conductivity (junhos/cm) 262 262 258 259 261 257 256

_______Temperature CC) 24.9 24.9 25.0 24.8 24.9; 25.1ý 24.8 25.1 24.9 20 24.9 24.9 25.0 25.2 (mg/L PH(SU) 7.89 8.05 8.07 7.94 7.851 8.081 8.09 8.03 7.96 89 7.84 8.07 8.06 7.98 Conductivi 86.4%

DO (mg[L)

(mg/L 7.8 7.8 7.8 7.9 7.8 7. 7.8 7.7 77.0 6 7.8 7.61 7.71 86.4% Conductivityrature fjsmhos/cm)

Residual 205 207 205' 2046 210' 203 201

_______Temperature CC) 24.9 25.1 25.0 24.9 25-0 2. 24.9 2 24.9 250.20 24.9 25.01 248 pH (SU) 7.901 8.051 8.05 7.3 7.85 8.9 8.06 8.5 7.951 8.091 7.75 8.09 8.021 7.99 DO t012/L) 7.91 7.81 7.8 7.9 7.9 Condnctivi m

7.7179 7. . 8.01 7.7 7.8 7.71 7.

Conductivity (tuhos/cns) 188 19 1882 189 1891816 Alkalinity 100%

100% Hardness Alkalinity Residual (nsg/L CaCO ) 73 37 3

Hardnesserature (mgfL CaCO ) 77 77 Total 3 Total Residual Chlorine (mg/L) '01 <0.10 <0.1 Tem Intake

_______Temperature (OC) 25.01 24.81 25.01 24.91 25.01 24.81 25.1 249 25.01 25.11 24.81 25.01 25.11 2.

pH (SU) 7.91 8.05 8.081 7.9 7.891 8.101 88.0 80 7.991 8.121 7.871 8.101 8.041 8.22 DO DO (mg/L) 8.0 7.91 7.81 7.8 7.91 7.81 7 7. 7. 7.71 8.01 8.21 7.81 Conductivi 7.71 7.7 Conductivity (~Imhos/cm) 18 18 18 17186ý 100%

Alkalinity 1886 Alkalinity 73 37 100% Intake Hardness (mg/L CaCO 3

)

Hardness (mg/L CaCO ) 79 77 Total 3 Tem Residual Total Chlorine (mr/L) I <0.1I01 01

_______Temperature (C) 25.01 24.91 25.11 25.0 24.91 25.01 25.0 24.8 25.01 24.91 24.81 25.01 24.9 2.

File: sqn101_11081 lchem.xls Entered by: C. Johnson Reviewed by:

Page 6 of 7

ETS

1 - Tnlll~lS~I~q4aI~nc Species: Ceriodaphniadubia Date: 1l1-0i9 Client: TVA / Sequovah Nuclear Plant, Outfall 101 Daily Chemistry:

I Day I Analyst I Ak%-r Ulfrr Concentration IParameter pH (S.U.)

DO (mg/L)

Conductivity (pmhos/cm)

CONTROL Alkalinity (g CaC03/L)

Hardness (mg CaCOv/L) * -

Temperature (°C)

DO (mg/L) 21.6%

43.2% Conductivity Conductivity ze*O (larnhos/cm)

PH (S.U.)

____________ Temperature ('C) 1 pH (S.U.) wQ DO (mgIL) 86.4% Conductivity (jiihos/cm)_

Temperature (°C)

DH (S.U.) 1-90 DO (m/L) 1.11 Conductivity IQ--

&+/-mhos/cm)______

Alkalinity 100% (mg CaCO3/L)

Hardness (Mg CaCo3/L)

TR chlorine (mgfL) 0.i Temnerature (°C) a -2&.8 i .

pH (S.U.) I-.A1 DO (mg/L) IN0 Conductivity I £f1-100% Alkalinity Intake (mg CaCQ 3/L)

Hardness (mg CaCh 3/L)

TR chlorine (mg/L) 40..0 U

Temperature (°C) U U-.0

-initial 11 Final I Initial 11 Final I Initial 11 Final i SOP AT 1I - Exhibit ATI 1.2, revision 06-01-11 IntiAns: v i~q& Kelley E. Keenan Initials: _

iETS Species: Cerlodaphniadubia Client: TVA / SeQuoyah I I Nuclear Plant,mOutfall 101 Date: 11-&,111 Day I K

II Day : I I _ . _ * " "5 I 5 I Analyst I Concen-tratin jParameter pH (S.U.)

DO (mg/L) *,",

".* Wv

.v Conductivity (gmhos/cm) -I-CONTROL .[Alkalinity (m, CaCOUL&

i _ _4 Hardness (mg TaCt3/L) *,

.,,-* . .4 Temnerature ('C) -. I nn(5~ TI~

PH (SU).

DO (ma*/U)

DO (mg/L) 10.8% Conductivity (iimhos/cm)

Temperature (*C) a.

- I - - I*~-Ii-nlli (* TT (S.U.) U DO (mg/L) 21.6% Conductivity -r 1-(umhos/cm)

Temerature (°C) 01.

IpH (S.U.) 0.10, I DO (mg/b) 43.2% Conductivity (gTmhos/cm) F1,3_

S.0

_________Temperature ('C) 1-njH (*q.TTa ).

pH (S DO (mr_/LI DO (mWL) 86.4% Conductivity (gtmhos/cm) a.

Temnerature (°C)

-emperaure - *C nit (Q TT "a PH (SU).

DO (mp/L) -'*" ;

I * . .. .. .. " "

Conductivity (jmhos/cm)

Alkalinity 100%

Hardness I I TR Chlorine (mg/L)

Temoerature (0 )

pH (S.U.)

DO (rag/L)

I Conductivity (jimhos/cm)

Alkalinity 100%

Intake (Mg CaCO3/L)

Hardness (mg CaCO3/L)

TR chlorine (mg/L)

! m I-I Temperature (QC)

I I

Initial II Final J SOP ATI1 - Exhibit ATL 1.2, revision 06-01-11 Paae 52 of 102 Kelley E.Keenan n pendert reviewby Initials:W

Page 1 of 6

ETS

, EnvlronmentalTesting Solutions. Inc.

Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephalespromelas JW*%I Client: Tennessee Valley Authority County: 4eA A-*.vb*J Facility: Sequoyah Nuclear Plant Outfall: 101 NPDES #: WN0,0269A 114 0 ,7J.0 Project #: -'"AV Dilutionpre arationinformation: Comments:

Dilution prep (%) 10.8 21.6 43.2 86.4 100 . Each concentration was UV-treated Effluent volume (mL) 270 540 1080 2160 2500 for 2 minutes to remove pathogenic Diluent volume (mL) 2230 1960 1420 340 0 Interferences.

Total volume (mL) 2500 2500 2500 2500 2500 Test organism information:

Test information:

Organism age: noLAN Randomizing template: &.0l Date and times organisms t -0"1- 1106 .0 Incubator number and were born between: shelf location:

Organism source: A,1W U-rck aci-k".i Artemia CHM number:

Drying informationfor weight determination:

Transfer bowl information: pH = 1. 1 C0 S.U. Date / Time in oven: IVOU 0 0

Temperature = " ' IC Initial oven temperature: t' Average transfer volume: Date / Time out of oven: "

0. I ' l , - Final oven temperature:

Total drying time:

Dailyfeeding and renewalinformation:

Day Date Morning feeding Afternoon feeding Test initiation, Sample numbers used MHSW renewal, or batch termination used Time Analyst Time Analyst Time Analyst Outfall 101 Intake 0I I I-Og-110 ,I

.0 0 l*.

W -#I At

A .. 1 1 iqoo A .s

. Vz. 7o A ,. lnI 1 0. 01 j luai**, "- IV0411A 1 ,I -,o'i ohoo j- '4 to0 t\LS< ' A1 "%% l nl '. o t*

.. ,,,t0,.o t 1*i ,Oa.

-L llo q-6+

11 4AL.

,\, l.-o.. il -*o -il 6

5__

_ _ _ -_ -__ _ O ..... L , . .. L . .4N _,Is i.t1 1111 11.16, 11-o0 0 1V14- d Al. WL

\I. M' I t ,I4 ii11111 I It-01-lI IL200 :ý Con trol in formation: \ ........ Acceptance criteria S u mm ary of test endpoints:

% Mortality: np7. s 20% 7-day LC5 o .. 07.

Average weight per initial larvae: 6.124'. NOEC 1007.

Average weight per surviving larvae: 0,124 1 0.25mg/larvae I LOEC >1007.

I ChV IC25 1 )1007 ,

SI PINAVen Initials: _ _

r yew'Kelley E. Keenan SOP AT20 - Exhibit AT20.3, revision 04-01-09

Page 2 of 6 "ETS EnvironmentalTesting Solutions, Inc.

Species: Pimephalespromelas Client: TVA / Seauovah Nuclear Plant. Outfall 101. UV-treated Date: ,1-CA-11 Survival and Growth Data Day CONTROL 10.8% 21.6%

A B C D E F G H I J K L o0 IQ /0 / 0 /0 /0 / 0 10 /0 /0 (0 /0-10 10 /0 (,) /0 /o /0 3 0 60 /0C I*o j 10 10c 10 10 1") 10 4ID 10 to /0 / 10 /M /I 1(3 t0. /6 (0 5 10 10 /o 10 .1 I0 . 10 lo 10 /0 (o 6

60 (0 10 /0- 1 ID0 10 /0 /0 /0 /0 /0 7 I0 10b f o / 0 1* ,ý-t o . IO I / to A = Pan weight (mug)

Tray color code:: AL*,,,.. .

Analyst: fA SI. 13.51 iZA'J 14.91S 16A .1 I l.l 1,.t 123.S'o.. 1'/4 Date: 1.t*lk.-L C = Larvae weight (mg) = B - A "hi( I' . &" I a.21 - 6.. i. q1'4 1 .2.(e 6 *S*1. 22.i 1 .S/ ,. .1 -

H and calculated .

Analyst: .. - -/- -

Weight per initial number of larvae (mrg)

= C /Initial number of larvae Hand calculated.

Analyst:

rio 0* 0' 0* 0

~0 tY

~0 4ý*L"t3%

01 01 0*

AA" No a * - * -

h Pa I Average weight per-initial number of Percent reduction from control () I 0 o.

2i (.617- 1..'7. 0.101 larvae (mug) I . a Comment codes: c = clear, d = dead, fg = fungus, k = killed, m = missing, sk = sick, sm = unusually small, Ig = unusually large, d&r = decanted and returned, w = wounded.

Comments:

SOP AT20 - Exhibit AT20,3, revision 04-01-09 Pl**n& *nt~evl *y Kelley E. Keenan Initials: J

Page 3 of 6 I

0ETS

EnvZonmentaITtng Solutions, nc.

Species: Pimephalespromelas Client: TVA / Sequoyah Nuclear Plant, Outfall 101. UV-treated Date: t1"O4-)l Survival and Growth Data Day 43.2% 86.4% 100%

/Io 10 /D 10 10

/ /0 10

10) o 10JO c 10 Jto '0 /0 ,0o to 10 ID /

2 _ /0/0 i C /0 C I00 /D 0 10 /0 10f0 23/ /OD 10O ' 0 1r IO to IOC (O 10 /I (

4 to /( 0 /D / /0 Ib I0 VIA lo /0 /0 10 5 it' 1(0 10 .tO C D) I ) O ( /( /0 1D 6 /0 / /0 /0. /ol0 / o 00 /0 C0 I f3 1(jC) t to A0.- 1011 /D /G I o 7

o' A - Pan weight (mg)

Tray color code:: l aA zt.

Analyst: ___ __1_._2_ i1__ 4 .' - is.0e, ls-'t.s "&&L, 1 I4.0 AS 4.01t 14.1A I.0. 13.-.57 Date: I01..\

B = Pan + Larvae weight (mg)

M4 It-l* . A . .. L.. t. 6.A Z.

Analyst:

1.1"'11 t -5uI &ejs Date:

C = Larvae weight (nmg) = B - A Hand calculated. ".O . 1.'* i,.22. 1,V1 t,.24 , " . "I Analyst:

Weight per initial number of larvae (rag)

= C / Initial number of larvae (7 X

'O "

Hand calculated.

Analyst: *{* O" r"

' q* '")" ("o " " )'

\ r-

- - %19

-A-. -....

Average weight per Percent reduction O,1Zt 0.'*7..i7. .

initial number of from control (%)M . ' 7.

larvae (mg) .....

Comment codes: c = clear, d = dead, fg = fungus, k = killed, m = missing, sk = sick, sm = unusually small, Ig = untsually large, d&r = decanted and returned, w = wounded.

Comments:

SOP AT20 - Exhibit AT20.3, revision 04-01-09 PiRp AnRnv9?y Kelley E. Keenan Initials: U

I Page 4 of 6 "ETS

.0 EnvlronmentafTesting Solutions. Inc Species: Pimephalespromelas Client: TVA / Sequoyah Nuclear Plant, Outfall 101, UV-treated Date: I-04I-11 I Day 100% Intake

_ __Y Z IAA BB 0

o 10// 0 tO00o 1

/D0 2 /0 3/(

4/ /0 5 to 1 o 6

7 /0 A f Pan weight (mg)

Tray color code::

Analyst: *s lkJ 1.50 Date: tO4*-*t2 B = Pan + Larvae weight (mg)

Analyst: _o 2_Ot ?4 . (.q Date:

C = Larvae weight (mg) = B - A Hand calculated. . "1.-1 q ,

Analyst: _ Al',

i. Weight per Initial number of larvae (mg)

= C / Initial number of larvae Hand calculated. d 4.U Analyst: D. 0.

Average weight per Percent reduction initial number of from control (%) 0, 1 1 ""7.

larvae (m_______ ___ ____

Comment codes: c = clear, d = dead, fg = fungus, k = killed, m = missing, sk = sick, sm = unusually small, Ig= unusually large, d&r = decanted and returned, w = wounded.

Comments:

SOP AT20 - Exhibit AT20.3, revision 04-01-09 l n ntTlvin Kelley E.Keenan Initials: ___=__

I ,

TVA / Sequoyah Nuclear Plant, Outfall 101 UV-treated November 08-15, 2011 EL PimephalespromelasChronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses toEnvironmental Testing Solutions, Inc. Prloect numbaer 7477 SNotfor Compliance Aýssessrmnente.at Laboratlr

'a MeweightI Cdfideatetvalfatloa Weight/ blitniamber Me rslvat Me-welghtlInial Coerlent or Perceni .dsajl froai Co.o-tea- (%) Re8plite Initial ...

ber o F1%t.J mbr, d-nrhse A P-.eight (ow B - Pa. + Ls'te Lt.-- weihtht(ow Weight I Survlvi8 hwe. weight (gia) -A--B .umbhrdtUaes (mg) Sursl.Z.$..aberia - .dlary.(ma0 (%) a..,tem tar,..rve,.ailtta tsa.., au atasdl (%)

kres(e* '" I)(s A I0 I0 13.78 20.59 6.81 0.681 0.681 21.07 7.18 0.718 0.724 4.7 0.718 100.0 0.724 4.7 Not applicable Control B 10 10 13.89 0.733 0.733 Control C 0 10 12.95 20.28 7.33 D 10 10 14.85 22.47 7.62 0.762 0.762 E I0 10 13.62 20.50 6.88 0.6880.688 F 10 10 13.69 19.95 6.26 0.626 0.626 6.47 0.647 4100.0 0.672 6.6 7.

G 10 10 13.23 19.70 H 10 10 14.71 21.97 7.26 0.726 0:726 t o0 10 13.93 20.48 6.55 0.655 0655 21.98 7.29 0.729 0.703 6.7 0329 100.0 0.703 6.7 2.8 21.6% 10 O 10 14.69 K 10 t0 12.75 20.31 7.56 0.756 0.756 L to o0 14.03 20.75 6.72 0.672 0.672 M 10 10 14.92 21.98 7.06 0.706 0.706 N 10 10 14.37 21.76 7.39 0.739 0339 0 10 10 13.08 19.90 6.82 0.682 0.682 P 10 10 13.75 21.32 7.57 0.757 0.757 9 10 10 15.62 21.83 6.21 0.621 0.621 20.56 6.48 0.648 0682 11.3 97.5 0.665 133 8-1 R 10 10 14.08 S 10 10 14.85 22.79 7.94 0.794 T t0 9 14.07 20.04 5.97 0.663 0.597 U 10 10 14.29 22.64 8.35 0.835 0.835 13.32 20.08 6.76 0.676 0714 11.3 676100.0 0.714 113 13 100% V 10 10 0.665 W 10 10 13.97 20.62 6.65 0.665 X 10 10 13.52 20.33 6.81 0.681 0.681 Y 10 10 13.17 20.61 7.44 0.744 0.744 21.05 8.21 081 0.761 6.4 100.0 0.761 6.4 -5.2 100% Intake Z 10 10 12.84 AA 10 10 13.75 21.49 7.74 0.774 0.774 BB 10 1 10 14.50 21.56 7.06 0.706 0.706 outrian 101; MSD = Minimum Significant Difference Dunnetf's MSD value: 0.1004 PMSD = Percent Minimum Significant Difference PMSD: 13.9 PMSD is a measure of test precision. The PMSD is the minimum percent difference, between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Intake:

Duonett's MSD value: 0.0575 Lower PMSD bound determuned by USEPA (10th percentile) = 12%.

PMSD- 7.9 Upper PMSD bound determined by USEPA (90th percentile) - 30%.

Lower and upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET Intehraboratuory Variability Study (USEPA, 2001a; USEPA, 2001 b).

USEPA. 2001a, 2001b. FinialReport: Interlaboratory Variability Study ofEPA Short-tam Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes I and 2-Appendi EPA-821-1-01-004 and EPA-821 -B-01-005. US Environmental Protection Agency, Cincinnati, OH.

Fie: sqnlO1_11O811data-uv.xlsx Entered by: J. Sumaer Reviewed by:

TVA / Sequoyah Nuclear Plant, Outfall 101 - Intake UV-treated November 08-15, 2011 I"ETS Statistical Analyses Env nmenti Testing r Larval Fish Growth and Survival Test-7 Day Growth Start Date: 11/8/2011 Test ID: PpFRCR Sample ID: TVA / SQN, Intake End Date: 11/15/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring Report Sample Date: November 2011 Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelas Conc-% 1 2 3 4 D-Control 0.6810 0.7180 0.7330 0.7620 10.8 0.6880 0.6260 0.6470 0.7260 21.6 0.6550 0.7290 0.7560 0.6720 43.2 0.7060 0,7390 0.6820 0.7570 86.4 0.6210 0,6480 0.7940 0.5970 100 0.8350 0.6760 0.6650 0.6810 Intake 0.7440 0.8210 0.7740 0.7060 Transform: Untransformed 1-Tailed Conc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD D-Control 0.7235 1.0000 0.7235. 0.6810 0.7620 4.659 4 10.8 0.6718 0.9285 0.6718 0.6260 0.7260 6.609 4 21.6 0.7030 0.9717 0.7030 0.6550 0.7560 6.747 4 43.2 0.7210 0.9965 0.7210 0.6820 0.7570 4.646 4 86.4 0.6650 0.9191 0.6650 0.5970 0.7940 13.306 4 100 0.7143 0.9872 0.7143 0.6650 0.8350 11.309 4 Intake 0.7613 1.0522 0.7613 0.7060 0.8210 6.383 4 -1.277 1.943 0.0575 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.97801 0.749 0.07415 -0.6805 F-Test indicates equal variances (p = 0.56) 2.07766 47.4683 Hypothesis Test (1-tall, 0.05) MSDu MSDp MSB MSE F-Prob df Homoscedastic t Test indicates no significant differences 0.05746 0.07942 0.00285 0.00175 0.2489 1, 6 Treatments vs D-Control Dose-Response Plot 0.9 0.8 0.7 ......... 1-tail, 0.05 level

...... ...... ............ of significance 0.6 0.5 09'0.4

0.3 0.2 0.1 0_

1CO4 o -O B C

File: sqnl0l_110811data-uv.xlsx Entered by- J. Su ner Reviewed by:

PMVW~~~ Kelley E.Keenan I, Initials:____

TVA / Sequoyah Nuclear Plant, Outfall 101 UV-treated E November 08-15, 2011 TStatistical Analyses "E

nmeTeng eAioen, tm.

Larval Fish Growth and Survival Test-7 Day Growth Start Date: 1118/2011 Test ID: PpFRCR Sample ID: TVA I SQN, Outfall 101 End Date: 11/1512011 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring Report Sample Date: November 2011 Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelas Conc-% 1 2 3 4 D-Control 0.6810 0.7180 0.7330 0.7620 10.8 0.6880 0.6260 0.6470 0.7260 21.6 0.6550 0.7290 0.7560 0.6720 43.2 0.7060 0.7390 0.6820 0.7570 66.4 0.6210 0.6480 0.7940 0.5970 100 0.8350 0.6760 0.6650 0.6810 Intake 0.7440 0.8210 0.7740 0.7060 Transform: Untransformed " 1-Tailed Isotonic Conc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Mean N-Mean D-Control 0.7235 1.0000 0.7235 0.6810 0.7620 4.659 4 0.7235 1.0000 10.8 0.6718 0.9285 0.6718 0.6260 0.7260 6.609 4 1.242 2.410 0.1004 0.6986 0.9656 21.6 0.7030 0.9717 0.7030 0.6550 0.7560 6.747 4 0.492 2.410 0.1004 0.6986 0.9656 43.2 0.7210 0.9965 0.7210 0.6820 0.7570 4.646 4 0.060 2.410 0.1004 0.6986 0.9656 86.4 0.6650 0.9191 0.6650 0.5970 0.7940 13.306 4 1.404 2.410 0.1004 0.6896 0.9532 100 0.7143 0.9872 0.7143 0.6650 0.8350 11.309 4 0.222 2.410 0.1004 0.6896 0.9532 Intake 0.7613 1.0522 0.7613 0.7060 0.8210 6.383 4 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.88775 0.884 1.11844 0.85946 Bartlett's Test indicates equal variances (p = 0.44) 4.78848 15.0863 Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnetrs Test . 100 >100 1 0.10042 0.1388 0.00258 0.00347 0.60095 5, 18 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point % SD 95% CL(Exp) Skew IC05 >100 IC10 >100 IC15 >100 1.0

>100 0.9 IC20 I1C25 >100 IC40 >100 o.a IC50 >100 0.7 0.6 C

o 0.5 0.4 0.3 0.2 0.1, 0.0 . . .

0 s0 100 150 Dose %

Dose-Response Plot 0.9,-

0.8 0.7

...--- ..--..-- ..-.-- .-....--------- 1-tail, 0.05 level 0.6 iof significance 0.5i 0

" 0.3 0.2 0.1 0I COC File: sqnl0IO 11081data.-uv.xlsx Entered by: J Siner Reviewed by:_

PI~nAA vi,9?y Kelley E. Keenan Initials:V'

TVA / Sequoyah Nuclear Plant, Outfall 101 - UV-treated November 08-15, 2011 Pimephalespromelas Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Daily Chemical Analyses 7 Environmental Testing Solutions, Inc. Project number: 7477 m m Concentration Parameter nH tSIT1 I 4-.

DO (mLILI Conductivitv turnhos/cm)

Control Alkalinity (mgfL CaCO3 ) L Hardness (mng/L CaCO)

_______Temperature C'C) 25. U 4 2:5.U 4 24.9 24.81 5. 24.81 24.9 24.b 24.7 24.51 24.61 24.5 PH (SU 7.89 78 8.03 770 .89 7.7 8.07 7.871 7.98 7.71 8.01 7.791 8.031 7.77 DO (mgtL) 7.8 8 8.0 779 6.9 7.7 7 7.7 6.8 8.0 76.77 753 1.% Conductivity (prnhos/cm) 305 305 298__ _____ 291______

_______Temperature CC) 24.9 247 24.9 2418 25.0 249 24.9 25 25.0 24.5 24.7 24.5 24.8 24.8 pH(SU) 7.89 7.91i 8.04 7.73 7.89 7.8 8.06 70 7.98 7.74 7.99 7.80 8.04 7.79 DO ( gL) 7.8 78 8.0 7.7 7 7. 0 7.7 78.7 7.0 8.0 7.6 7.7 7.61 21.6% Conductivity (grnhos/cm) 290 290 284_25_28 284 ~ 282

______Temperature CC i 25.0 24.7 25.0 24.8 25.0 2 925.0 24.5 25.0 24.5 24.8 24.71 24.8 24.7 PH (SU) 7.91 7.89 8.04g 7.72 7.88 73 8.06 7.89 7.98 7.70 7.94 7.871 8.04 7.77 DO (mg/L) 7.8 7.7 7.9 7.7 79. 9 7.7 7.8 7.8 7.1 8.1 7.61 7.61 7.6 432 Tonductivit prhos/cm) 262 261 260 29 25 259 259

______Temperature CC) 25.0 24.8 25 *0 24.6 250. 20 25.0 24.6 25.1 24.5 24.9 24.7 24.8 24.7 PH (SU 7.92 7.93 8.05 7.75 7.87 7.78. 8.05 7.95 7.98 7.87 7.87 7.91 8.03 7.84 8.% DO (mgL) 7.8 7.7 7.9 7.7 7.9 6.91 7.8 7.8 7.8 7.0 8.1 7.5 7.6 7.6 864% Conductivity (tumhos/cm) 211 211 ý ý 272520 ý203 206

_______Temperature (cC) 25.1 24.6 25.0. 24.6 25.0 248 25.1 24.6 25.1 24.5 24.9 24.7 24.8 24.4 PH (SU) 7.91. 7.90. 8.041 7.79 7.83 7.9 8.02 7.93. 7.96 7.6 7.821 7.901 8.031 7.82 DO (mgfL) 7.91 7.71 7.91 7.7. 7.9 70 7.9 7.81 7.9 728.01 7.51 7.61 7.7 Conductivity (jsmboslcm)191218 .11 10 100% Alkalinity (ingIL CaCO3 ) 7 Hardness (mgfL CaCO3)7977 Total Residual Chlorine (mg/L) <0.10 <0. I <0.10

_______Temperature CC) 25.1 24.61 25.21 24.51 25.11 24.91 25.21 24.7 25.2 245 24.91 24.61 24.91 24.6 pH (SUJ) 7.92 7.931 8.051 7.761 7.861 7.811 8.061 7.97 7.98 7.T7911 7.881 8.061 7.831 DO (mg/L) 7.9 7.71 7.91 7.61 7.91 7.11 7.91 7.8 7.9 718.21 7.51 7.71 7.5 Conductivity (junhos/cm) I9 ý191416 100% Intake Alkalinity (mgfL CaCOj) 77 7 CaCO 3 ) - WL (mng/LChlorine Total Residual <0.1 <0.10 lHardness

_______Temperature CC) 24.91 24.61 25.11 24.51 25.01 24.9 25.0 244 2. 45 24.91 24.61 24.9 24.6 File: sqn 101_110811chem-UV.xls Entered by: C. Johnson Reviewed by:

II Page 5 of 6 I 00ETS

Environmental Testing Solutions, Inc.

Species: Pimephalespromelas Date: 11i04-1 Client: TVA / Seauoyah Nuclear Plant, Outfall 101. UV-treated Daily Chemistry:

Day 0 1 ýZ 22 Analyst 0-0 1IODT I CW ICOT 11 IeD CW 11_m Concentration IParameter PH (S.U.) 4.9r4- *+W) -I-OMM- 11q-.1-3 I *A-A 11 431H DO (mr/Li Conductivity CONTROL (jimhos/cm) 51 Alkalinity UV-treated Hardness (mg CaCO3/L) ____

Temperature (°C) Ts. 0 pH CSU..IA 10.8% Conductivity (jgmhos/cm)

Temperature (C)Q t~'

21.6% Conductivity 021o SI (4.mhoscm)

___________Temperature (0 C) ).S .0-1 .0 4 .- . 4:.

I

,-:th -. ot (0di 43.2% Conductivity (pgnhos/cm) ~ILU 1110 0 1

_________Temperature (QC IIS .0 R I m.o 7AL- I 2S0

'M !".o I 86.4%

pH (S.U.)

DO (mg/L)

Conductvity Temperature (°C) 'L. I.

Conductivity (g+/-mhos/cm)

Alkalinity 10M% C 3 /0 Hardness (Mg CaCO 3IL)

TR chlorine (mg/L)

____________ Temperature ( C 0

Z.

Conductivity (jimhos/cm) 100% Alkalinity i" Intake (mg CaCO 3/L)

Hardness (Mg CaCO3/L)

TR chlorine (mg/L)

Temperature (°C) 2.

Initial I SOP AT20 - Exhibit AT20.3, revision 04-01-09 nIMA 91 & Kelley E. Keenan Initials:

Page 6 of 6 0

"DnETS I1 EvrnmetlTsig olutions, tne..

Species: Pimephalespromelas Client: TVA / Seauovah Nuclear Plant. Outfall 101. UV-treated Date: il-A~RII

i. ~~ ~ ..~]..... ~ ~ ]. ~ ...-~ ~,... ~ ~ ae 11..

.I?-OS "!

1 Day I Analyst 3L4. '

6 11 A M Concen- Parameter tration I pH (S.U.)

DO (rng/L)

Conductivity I(*imhos/cm CONTROL I UV-treated Alkalinity

(*m CaCOe/YU Hardness (me CaCOSL'i CaC03/1-)

iI (Mg Temperature (°C)

I j ._._ _

10.8% Conductivity I -

Temhos/cm) emperature C0 C)TU C

1H (S.U.)

I 9 -~--I: 7' --

DO (mrg]L) 21.6% Conductivity (umhos/cm)

Temperature (°C)

I U pH (S.U.)

DO (mg/L)

In S~

a. I1~-

43.2% Conductivity (Qmhos/cm) ,.wL..

Temnerature (°CM

________ - I 'i~ ii-pH (S.U.) - - -

DO (mg/L) 86.4% Conductivity (jimhos/cm)

Temperature (°C)

S U - -

pH (S.U.) I -

DO (mo/L) 9-Conductivity

(;tmhos/c.)

Alkalinity 100% (mg CaCO3/1.)

Hardness (mig CaCO3IL)

TR Chlorine (mg/L)

Temperature (0 C)

I r " " I - I II .. : ii .- 7 pH (S.U.) I= %.k II - : I ,'9,1 II T'.W I 9F.F - II q'*> I DO (mg)L)

. ..--- ~~~~~

I ~ +- ; kT-if 1 Conductivity (gmhos/cm) -'5 100% Alkalinity Intake (mg CaCO 3/L)

Hardness ieI (mig CaC03/L)

TR chlorine (nmg/L)

Temperature (°C) 2~.l N. 6 1 7.4L21 Initial Fina-l Initial FnlI Initil Thminal SOP AT20 - Exhibit AT20.3, revision 04-01-09 PIM enq-nPrSv(ea'y Kelley E. Keenan Initials: -i

Page 2-Page I of I e0ETS

" Environmental Testing Solutions, Inc.

Total Residual Chlorine (Orion Electrode Method, Orion 97-70)

Matrix: Water, RL = 0.10 mg/L Meter: Accumet Model AR25 pH/Ion Meter Ic Analyst F7 T-- IIIII,Iio 1

Acid reagent:1 odide I Date analyzed - reagent: TOI=L4, Calibration:

0.10 mg=L, 1.00 mgaL s S lope

/ .N$-O

! MRe~ference standard number I Arote: For samples with a residual chlorine of > 1.0 mg/L. the calibration range must be adjusted to bracket the chlorine levels of the samples.

Laboratory control standard:

Reference standard True value (TV) Measured value (MV) % RS = MV / TV x 100 number j (mg/L) (mgfL) (acceptable range = 90 to 110%)

0.50 044 riI Duia~;percso:Sampple characteristics R **n acetberng 70 Sample m peasurements:

Sample Sample ID Sample characteristics Residual chlorine number (mg/L) (

Reagent Blank L o.oRL 11 ND0Cfl rn Loe o7 2 I 111~.06 PCs,Dmsdhiae. Jxor'. 204a 001%

I nnoio1 1VA S %n 101 LAX_______

111l1(.CTZ-3 640 1q~4ctkt r-ke~,u- pe k Z'.0._0Q__3ayr Note: All samples were analyzed in excess of EPA recommended holding time (15 minutes) unless otherwise noted.

Laboratory controlstandard:

Reference standard True value (TV) Measured value (MV) % RS =MV / TV x 100 number (ragIL) (mg/L) (acceptable range = 90 to 110%)

!-U )tZ , 0.50 .).,, o57 °1 Reviewed byI V Date reviewed Page 63 of 102 SOP C8 - Exhibit C8.1, revision 06-01-11

Page t Page i of_

S EnTS Inc.

41Environmental Testing Soluflonrn Total Residual Chlorine (Orion Electrode Method, Orion 97-70)

Matrix: Water, RL = 0.10 mg/L Meter: Accumet Model AR25 pH/Ilon Meter Analyst I's I odide reagent: I ' 7 I.4 : : :

Date analyzed 10. It Acid reagent: I*,I /o I Calibration:

0.10 mg/L Loo mgL I Slope Roeference standard number -tD t '

Note: For samples with a residual chlorine of > 1.0 mg/L, the calibration range must be adjusted to bracket the chlorine levels of the samples.

Laboratory controlstandard:

Reference standard True value (TV) , Measured value (MV) % RS = MV / TV x 100 number (ing/L) (rag/L) (acceptable range = 90 to 110%)

0.50 Duplicatesample precision:

Sample SampleSEI)e Sample characterstic Residual chlorine %RPD = {(S - D) /[(S+D)/2]) x 100 number (mg (acceptable range = + 10%)

/Duplicate DZ Sample measurements:

Sample Sample ID Sample characteristics Residual chlorine number _m/L Reagent Blank Z o 12'$ .

\11 I.~O."L+/-. IA-9..LlM *X,.w a,...t4,[,-

rio,~x oo Note: All samples were analyzed in excess of EPA recommended holding time (15 minutes) unless otherwise noted.

Laboratory control standard:

Reference standard True value (TV) Measured value (MV) % LS)-7MV/TVx number (mg/L)I (IgL) (acceptable range = 90 to 110%)

ffl ti T.- KF 0.50 G O.S t6. o401.

Reviewed by I Date reviewed I Ib- II Page 64 of 102 SOP C8 - Exhibit C8.1, revision 06-01-11

Page _1 Page Iof ?.

ETS iNP XIi 1 Environmental Tsting Solutions, Inc.

Total Residual Chlorine (Orion Electrode Method, Orion 97-70)

Matrix: Water, RL = 0.10 mg/L Meter: Accumet Model AR25 pH/Ion Meter Analyst I odide reagent: LI*.* IIZI Date analyzed . Acid reagent: I-S-4,4o Calibration:

1 I 0.10 mgaL 1.00 mglL 'Slope Reference standard number U %NMA1," 7-1.

Note: For samples with a residual chlorine of > 1.0 mg/L, the calibration range must be adjusted to bracket the chlorine levels of the samples.

Laboratorycontrol standard:

I [ Reference standard

-4 number True value (TV)

(mg/L) 0.50 Measured value (MV)

(mg/L) o.0,, c

% RS = MV / TV x 100 (acceptable range = 90 to 110%)

.o Duplicate sample precision:

Sample Sample ID Sample characteristic's Residual chlorine %RPD = ((S - D) I[(S+D)12]j x 100 number II (mg/Li (acceptable range = 1 10%)

Duplicate D 40. 00- 01961 Sample measurements:

Sample Sample ID Sample characteristics Residual chlorine number _m__L_

Reagent Blank 610.II__

I I I JL . 7- 001~ ,i e'"k 1 CA" 4-0_0004(

0.2000% 4-A A lilt 14, icp-uit peg.~ a..r 411cY4 l~llLIL ~0. 4 , 6.-. _____1 0.2 cor).-L14 AM 0351 Note: All samples were analyzed in excess of EPA recommended holding time (15 minutes) unless otherwise noted.

Laboratorycontrol standard:

-Re nce standard True value (TV) Measured value (MV) % RS = MV / TV x 100 I number ( /) (mg/L) (acceptable range = 90 to 110%)

0.50 L. j Reviewed by I Date reviewed I- IT12-Page 65 of 102 SOP C8 - Exhibit C8.1, revision 06-01-11

., S 0

Page 81 Page ___j of 5 Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO 3/L Analyst f" 7,1- 7 Time initiated j[qq Date analyzed Titrate samples to Time completed IS I pH = 4.5 S.U.

..... * ............ J_." ....

Duplcatesampe precision: .

Sample Alkalinity %RPD Sample Sample ID volume Begin End Total IMultiplier (Ing CaCO3/L) [(S - D) /[(S+D)/21) x 100 (ml) ml ml I ml.M / (acceptable range 110%)

number Duplicate(B) , I*6M 3.z- , ° "32.. 5.r/

Matrix spike recovery:

Reference standard Spike value Sample Spike alkalinity (A) number (SV) volume Begin End Total Multiplier (01g CaCO3/L).

(mg CaCO3/L) (ml) ml ml ml (ing CaCO3/L)

I Sample alkalinity (B) Measured spike value (MV)

MV = A - B (mg CaCO3/L)

% R= MV / SV x 100 (acceptable range

-75 to 125%)

I Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID (ml) ml ml ml Multiplier (mg CaCO3 /L) t.*m- %\ _15%,Z1 63S 3,4,. 10 3.* /

R-0 13.o 0O0

__-_____I qo.O -(q. / 0 Z.

-\i., *... 5 (4,.I !oz-.

Al~~~~o*~~0- _ ~. 6-1___ -z1 h 410 01-:Z. Z9.0 __ __ _ _ _0 "q" II Page 66 of 102 Reviewed by: E= Date reviewed: -

SOP C6 - Exhibit C6.1. revision 09-01-09

Page __

Page 2. of J; SEnvironmental Testing Soludons,on Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO 3/L Analyst 7Pkil'i,,- I Time initiated [Id..IL,, ]

Date analyzed Ii-,iq.I I I Titrate samples to Time completed [ ZZ-pH = 4.5 S.U.

Titrant nornmality and multinlier determinatioi PH of Normality Normality (N) of H2 SO4 pH Factor or Multiplier Deionized Titrant check Begin End Total = (5 ml Na 2 CO3 x 0.05)/E = (N x 50000)/100 ml sample water reference standard ml ml ml = 0.25/E = N 500

=

= 4.5 S.U. number number (E) (acceptable range = 0.0180 - 0.0220) _

Laboratory controlstandard:

Reference standard True value Sample Alkalinity (MV) % RS =MV / TV x 100 number (TV) volume Begin, End Total Multiplier (mg CaCO3IL) (acceptable range (mg CaCO3fL) (ml) ml ml ml = 90 to 110%)

I1 *'iS'i 100 100 Z.j.o 3& q,3- o7 Duplicatesample precision:

Sample Alkalinity %RPD =

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3/L) ((S - D) /[(S+D)/211 x 100 (ml) ml ml ml (acceptable range=L 10%)

number I- d5* jUgt. 6 .0 '44 'f-1 1) 10-1 *S M 4, Duplicate (B) j~,_____ D _______

Matrix spike recovery: __,

Reference standard Spike value Sample Spike alkalinity (A) number (SV) volume Begin End Total Multiplier (mg CaCO3 lL)

(Ing CaCO3/L) (ml) ml ml ml Sample alkalinity (B) Measured spike value (MV) % R = MV / SV x 100 (mg CaCO3/L) MV = A - B (acceptable range (mg CaCO3/L) = 75 to 125%)

-zoo Iq *0/

Sample measurements: - -

Sample volume Begin End Total Alkalinity Sample number Sample [D (ml) ml ml ml Multiplier (mR CaCO3/L) i-~'4iI~ ?.3 U-3 ___ __.0_____

1111jO,01U Z ,7.. _426 "7S 111112-01 o.,6 7.5 ,5s 7C I

I I1, I ,IIll11Z.

07--- Mitl~t VIC .7 IV* 1 1'.0 1-0 Date reviewed: I I\- 1411

! Page 67 of 102 Reviewed by:

SOP C6 - Exhibit C6.1. revision 09-01-09

.j.

Page 'If Page 3 of5 Environmental Testing Solutions, Ise.

Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO 3/L Analyst 3u Time initiated Date analyzed 11..11 Titrate samples to Time completed ]

pH = 4.5 S.U.

Titrant normaliy and multiplier determination:

pH of Normality Normality (A) of H2S0 pH Factor or Multiplier 4

Deionized Titrant check Begin End Total = (5 nil Na2 CO3 x 0.05)/E = (N x 50000)/ 100 ml sample water reference standard mi MI mi = 0.25/E ' . = N x 500

-4.5 S.U. number number (E) (acceptable rang$= 0.0180- 0.0220).

- *- " -. i " t 1 Laboratorycontrolstandard:

Reference standard True value Sample Alkalinity (MV) % RS = MV / TV x 100 number (TV) volume Begin End Total Multiplier (mg CaCOgL) (acceptable range (Ing CaCO3 /L) (ml) ml ml ml = 90 to 110%)

0191iq~ 100 10o aq.c 34.1, OI9 U.i.6%

Duplicatesa= e precision:

Sample Alkalinity %RPD =

Sample Sample ID volume Begin End Total Multiplier (mng CaCO 3/L) ((S - D)/[(S+D)/21} x 100 number (ml) ml ml. ml (acceptable range = E 10%)

111110,06 CItA 007Z- .50 '34-1 f4- ,10, 5 rl..S s). .0 Duplicate (B) 1 ,, ? 107D Matrix spike recovery:

Reference standard Spike value Sample Spike alkalinity (A) number (SV) volume Begin End Total Multiplier (rag CaCO3/L)

- (mgCaCO 3 /L) (ml) ml il mi l

%'tSIsal 10:0 so 0~ t,0i I'-5 $.J 16.1 l t i Sample alkalinity (B)

(mg CaCO3/L)

Measured spike value (MV)

MV = A - B

% R = MV / SV x 100 (acceptable range (mg CaCO3/L) = 75 to 125%)

ZZO q Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID (mi) ml mi ml Multiplier (Ing CaCO 3/L) lilito. 1 l M 001. UNWA ., 1*, UZ. 7.3 c2) to. _ ISO 1 1 . 1 "T' 4/P, I,,*. ~ A U.i 24 7 3,4 cz ) 7_

M120 Il~.' u,Uf*t-meAi 1, .74.2 ý,.3 U.; 73 11 i .0S I.TLATC. &A L.X "U3 S ( ,i) 2 C0 odeAO . 3 . L1 S.5 Itzo 1lllI0M,- .*0 ' ' c.Ij

r. (.0_)_

I

_1_ _ _

i 111 a1-,0S I'i A11.&olI ujteq eALor -LS Cit a(, i..q

'~aI.S I& C&1)

=o000 i Page 68 of 102 Reviewed by: I2ZZ Date reviewed: I 711-ii' SOP C6 - Exhibit C6. 1. revision 09-01-09

Page IZ 3

Page _ __of 5 nTeSo

) EnvironmentalTestIng Solutions, Inc.

Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO 3/L Analyst 374 7b Time initiated [ [

Date analyzed nI.i I ] Titrate samples to Time completed LZI * ]

pH = 4.5 S.U.

TifrnnI nn~rnmnlsu aNd inItlnfar ,Ln'..,.~itt..

pH of Normality Normality (N) of H-2 S0 4 pH Factor or Multiplier Deionized Titrant check Begin End Total = (5 ml NaZCO 3 x 0.05)/E = (N x 50000)1100 ml sample water reference standard ml ml ml = 0.25/E -= N x 500 4.5 S.U. number number (E) , (acceptable range = 0.0180 - 0.0220)

Laboratorycontrolstandard:.

Reference standard True value Sample Alkalinity (MV) %RS=MV/TVx 100 number (TV) volume Begin, End Total Multiplier (rag CaCO3/L) (acceptable range (mg CaCO 3/L) (ml) ml ml ml =90 to 110%)

Xts s ol' X 100 100 3 S .o.q g. q7 qT.0 Duplicatesam, le precision: Samle__ad___A]___________

Sample Sample ID volume Begin End Total ,NMultiplier (Ing CaCO3/L) [(S - D)/i(S+D)/21) x 100 number (ml *-*04 ml ml ml (acceptable range = k10%)

Duplicate (B) lam.. C,S f- DC Matrixspike recovery:

Reference standard Spike value Sample Spike alkalinity (A) number (SV) volume Begin End Total Multiplier (mg CaCO3/L) 1 (mg CaCO3 L)- ml mi "mml ml Xt-16s aj Soo 10 43 11,3- ,06 1 Sample alkalinity (B) Measured spike value (MV) % R = MV / SV x 100 (rag CaCO3/L) MV = A - B (acceptable range (mg CaCO 3IL) = 75 to 125%)

Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID m ) t.,% ml ml ml Multiplier (Ing CaCO 3/L)i 1.-1.4L I 1lL -o\

unnot oi T11- S7

-ix

~

+/- t7o% t.f 5W__

i, ,1 1)

-3.3

-w s1'~.

of io.&C i'.i 5.C It.C N i . C CIO)

CZI7 "6.)1w I I11o II nI U. .I I n o 0i.l ZI Ial. 67 -CIA,) loZ4

- 71Z r8 IC3.4 Iwo6 3.4

.'e.C 73 73 73 I *I 11aoI.22-iiI I .o tok t C

3*

o,¢ X'_'.9"l. 3.C 3*,l 5.

&¢ "73 73 I Reviewed by: Date reviewed: I 11.14-W/

Page 69 of 102 I SOP C6 - Exhibit C6.1. revision 09-01-09

S Page '13 Page .9 of 5

-* EnvironmentaliTesting Solutions, inc.

Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO 3/L Analyst

Time initiated Date analyzed ý l Titrate samples to Time completed pH = 4.5 S.U.

Tihrant nnriralift and ,nultlnlt'rde'iprmlnatln

-...... no pH of

.. and Normality inadol, Normality (A) of [12SO4 Tr pH Factor or Multiplier Deionized Titrant check Begin End Total = (5 ml Na2CO3 x 0.05)/E = (NI x 50000)l 100 ml sample water reference standard ml mI ml = 0.25/E = N x 500

- 4.5 S.U. number number (E) (acceptable range = 0.0180 - 0.0220)

Laboratory control standard:

Reference standard True value Sample Alkalinity (MV) % RS = MV / TV x 100 number (TV) volume Begin End Total Multiplier (Ing CaCO3/L) (acceptable range (Mg CaCO3/L) (ml) ml ml ml =90 to 110%)

gN5 116 100 100 31.4 41.,) CL; 1.1 * *G Duplicate sam le precision:

Sample Alkalinity %RPD =

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3IL) {(S - D) /[(S+D)/21) x 100 number (MI) MI mi. MI (acceptable range=-+ 10%)

Duplicate (B) S..* D - _;: S Matrix spike recovery:

Reference standard Spike value Sample Spike alkalinity (A) number (SV) volume Begin End Total Multiplier (Ing CaCO3IL)

(mg CaCO3 /L) (ml) ml i l ml

.X'* qsq 100 so S.O 11.0* 1.q ' 01I17 Sample alkalinity (B) Measured spike value (MV) % R = MV / SV x 100 (mg CaCO3/L) MV = A - B (acceptable range (mg CaCO3.L) = 75 to 125%)

73 j7 Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID (mi) ml ml ml Multiplier (rag CaCO3IL) 11110,1-.2.'2-, . , (-,L) .0 205 3.6 73 tL111l. li, .. S MILt 3.7 75 01o q'q q.q _5

______,_ / b .

OT.O__

-- * ' II.l~ - - -

Reviewed by:

z~IIz Date reviewed: [ I'I.I-jrlJ I Page 70 of 102 SOP C6 - Exhibit C6.1. revision 09-01-09

Page 73 Page I of .I Envronrne.nt

Testtng Solutions In.

Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO 3/L Analyst *i 1 Time initiated ois' Date analyzed 11.1 .11 Time completed rGo' Titrant normalitv and multiplierdetermination:

Titrant Normality check Begin End Total , Normality (N) of EDTA pH Factor or Multiplier reference standard ml ml ml = 0.2/E = (N x 50000)/50 ml sample number number (E) (acceptable range = 0.0180 -.0.0220) = N x 1000 Laboratory controlstandard:

Reference standard True value Sample Hardness (MV) % RS = MV / TV x 100 number (TV) volume Begin End Total Multiplier (mg CaCO3/L) (acceptable range (ligCaCO3/L) (ml) ml ml ml =90 to 110%)

40 50 16.1 I,1 2 . 0o I2. :00.8%

Duplicatesa le precision:

Sample Hardness %RPD =

Sample number Sample ID volume (ml)

Begin ml End ml Total ml Multiplier I

(mg CaCO3 /L)

Li

((S - D) /[(S+D)/2]) x 100 I ~

Matrix spike recover, :

Duplicate (B) Z. D0________

Reference standard Spike value Sample Spike hardness (A) number (SV) volume Begin End Total Multiplier (mag CaCO 3/L)

-f(ing CaCO 3/L) (ml) ml mi ml 0W "IIo 1 s qI i*Io Ill)4.

111 1. 1'7 Sample hardness (B) Measured spike value (MV) % R = MV / SV x 100 (rag CaCO3/L) MV = A - B (acceptable range (mg CaCO3/L) = 75 to 125%)

4o 3q ,010, Sample measurements:

Sample volume Begin End Total Hardness Sample number Sample ID (ml) ml ml ml Multiplier (Mag CaCO 3/L)

TV = ND Blank (should be = 0 Mg CaCO3/L) 0.I 0 ,0 ii'S D.o O.-0'\-1i A -I__1 Z,4.7 Zi.3 q.1 __.

1-VA- \ F6_ 21.3 '33.7 41.',

___-__ I3.1*. 35.Z St_ _.

___-__-_t_ 1__*_ __I * ,i4.7 Vi.5 ll 0-1d.11 A ~MOW, _____ 0.4 '.C Lt.5 11-0+1%~6 1.L ~ .. ___K_). Z

'ix '4- ___ _ 97 8

P tteef*4 &I102 ant isused, sample must be diluted. Reviewed by: Dtriwreviewed Date Ib . 0 -00 SOP C7 - Exhibit C7. 1. revision 09-01-09

Page ?q Page 2 of_ _

3 Environmentll Testing Solutions. In.

Total Hardness (SM 2340 C)

Analyst .E$ RL = 1.0 mg CaCO 3/L Time initiated '4 I77 Date analyzed ' 11.1-5. 1 III Time completed L ~

Titrant normality and multinlier determination:

Titrant Normality check Begin End Total Normality (A) of EDTA pH Factor or Multiplier reference standard ml ml ml = 0.2/E = (N x 50000)/50 ml sample number number (E) (acceptable range = 0.0180 - 0.0220) = Nx 1000 Laboratoy control standard:

Reference standard True value Sample Hardness (MV) % RS = MV / TV xI00 number (TV) volume Begin End Total Multiplier (Mag CaCO3/L) (acceptable range (mag CaCO3/L) (mi) ml ml ml = 90 to 10%)

5 ill 40 502 .J I1.O 1 .o C-0 '40 Sooo. Vo Duplicatesamwle precision:

Sample Hardness %RPD =

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3/L) ((S - D)/[(S+D)/2]) x 100 number ___ (__

ml) ml ml ml 11- 41.- A QQ s. i. '4.5 I"S[ j,lI Matrix spike recovery:

Reference standard Spike value Sample Spike hardness (A) number (SV) volume Begin End Total Multiplier (mg CaCO3 /L)

IM53s qtq (mg CaCO3/L) 40 (ml) 6o*0 Ml 15S.

ml ZIA1 CAq ml lies 130 j

Sample hardness (B) Measured spike value (MV) %R = MV / SV x 100 (mg CaCO3/L) MV = A - B (acceptable range (mg CaCO3/LL) = 75 to 125%)

Sample measurements: _

Sample volume Begin End Total Hardness Sample number Sample ID (ml) ml ml ml Multiplier (mg CaCO3/L)

TV- ,4D - Blank -,- ,

(should be = 0 mg CaCO,/L) -

I%IkC.06' 1 23 vi- 27U.57

, 'S _ .L I1111o0,03 a,.c7. C 3A7 r.I EI __ 10 I it,11OR. 01 J, So 31.4 .4.3 5.4 Ito.

I ______1 0-l

__, -\>. , (I') _ _o _

0. l

.'0{*5.4 I1.

. S13.4

&&1 1.5_

.110.

30 I it 11 to. 7'*

Ca

'64 ICA.Z IM1 1

.. 4 C7.,~ I p Pli1IFl 0A Mi ant is used, sample must be diluted. Reviewed by: Date reviewed SOP C7 - Exhibit C7.1. revision 09-01-09

Page 75 Page 3 of -4 EnlronmantilTestilng Solutions. Inc.

Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO 3/L Analyst I IIII Time initiated fýq' ]

Date analyzed 51-i3.-i. Time completedi Titrant normality and multiplier determination:

Titrant Normality check Begin End Total , Normality (N) of EDTA pH Factor or Multiplier reference standard ml ml ml = 0.21E - (N x 50000)/50 ml sample number number (E) (acceptable range = 0.0180 -.0.0220) = N x 1000 Laboratorycontrol standard:

Reference standard True value Sample Hardness (MV) % RS = MV / TV x 100 number (TV) volume Begin End Total Multiplier (9g CaCO3/L) (acceptable range

( Tg CaCO3/L) (ml) ml ml ml = 90 t o rag%)

% 111 40 50 1q.G t1.1 7.o) I11.S 14 too.O%

Duplicate sample precision:

F Sample Hardness %RPD Jinumber Sample 4

Sample ID Duplicate (B) volume

.A!!

Begin ml 0446~13?.

End ml Total ml Multiplier I10 (mg CaCO 3/L)

D 10

((S - D) /(S+D)/21) x 100 Matrix spike recovery:

Reference standard Spike value Sample Spike hardness (A) number (SV) volume Begin End Total Multiplier (Ing CaCO 3/L)

(mg CaCO3/L) (ml) ml ml ml

%N65 _*0It Io _q.q _7._ 7.3 1q. f MOD Sample hardness (B) Measured spike value (MV) % R = MV I SV x 100 (mg CaCO 3/L) MV = A - B (acceptable range (mg CaCO3/L) = 75 to 125%)

%to So0o0 7S.0O%

Sample measurements:

Sample volume Begin End Total Hardness Sample number Sample ID (ml) ml ml ml Multiplier (mg CaCO3/L)

T,+D Blank - - -i. ,1 lI I~w. O02-M o1.-,3--A i 5 S 3. 7.9 .,,.S0 1 itllL.0.6 DA .CF 00% _ 15 _ .1 40'l 3.9 1So it l10t.,:1 _., )ftv* &v*l qq~ v. 4.,, 93!

II-,i"L A;0 -.. _ z.6 3.1 3.A 77 fl0.2 nZ it" -4v 'sA 77 111j1F( tWr1PS 14J C% %117 117 '1.0 174 22-01,0'%., __ - 1__7 111.i*i 3.1 _[___

_ 77 F*glt'7~fMt"5 01nt is used, sample must be diluted. Reviewed by: [ Date reviewed 1 1 3"s-1l SOP C7 - Exhibit C7. 1. revision 09-01-09

Page 7C Page 9t of q

) Envifomental Testing Solutio Inc.

Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO3/L Analyst M Time initiated Date analyzed Ii.11i131 1 Time completed Titrant normality and multiplier determination:

Titrant Normality check Begin End Total , Normality (N) of EDTA pH Factor or Multiplier reference standard ml ml ml = 0.2/E = (N x 50000)/50 ml sample number number (E) (acceptable range 0.0180 n0.0220) -N x 1000 Laboratory controlstandard:

Reference standard True value Sample Hardness (MV) % RS = MV / TV x 100 number (TV) volume Begin End Total Multiplier (mg CaCO3 /L) (acceptable range (mg.CaCO3/L) (ml) ml ml ml =90 to 110%)

. "J5,lc40 50 01.6 lOo0%o Duplicatesam le precision:

Sample Hardness %RPD =

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3/L) {(S - D) /[(S+D)/2J} x 100 number . (ml) ml ml ml S DuplicateB "lt 3, D ,75-Sample volume Begin End Total Hardness Sample number Sample ID (ml) ml ml ml Multiplier (mg CaCO 3/L)

TV=*q B lank vi&%

(should be = 0 mg CaCOt/L R 1 It I%Mot -WKSW 10k tJ 1 so 31.2 35A '4.0 7 lil It A_ 7

_L&___6(R1__.

III 0t .,ft I111 2-23.-q (9 s*.- wrU' .T4 37, .9 3.q

lq75 77

__ 6______ 37.5 '4.1- f44.0 17 PAelt7*llM lfi)antis used, sample must be diluted. Reviewedby: ti .I* I Date reviewed ,- III SOP C7 - Exhibit C7.1. revision 09-01-09

Sequoyah Nuclear Plant Biomonitoring November 08 - 15, 2011 Appendix D Reference Toxicant Test and Control Chart Page 75 of 102

Pimephalespromelas Chronic Reference Toxicant Control Chart EiTS Organism Source: Aquatox, Inc.

(~Environmental Testing Solutions, Inc.

1.0 USEPA Control Limits (+/-2 Standard Deviations) 0.9 0.8 0.7

.............. . ° 0.6 0.5 1.4 I I I I I I I I I I I I I I I I I I USEPA Warning and Control Limits (75th and 90th Percentile CVs) 1.2 1.0 ......................................

M I_ 0.8 0.6 0.4 . ......................................................................

I I I I I I *I I I I I I I I I I i 1.4 ! I I I I I I I I I I I I I I I I I I I Laboratory Warning and Control Limits (10th and 25th Percentile CVs) 1.2 1.0 0.8 .................................... oo.'.............................o..........o...................o....

0.6 0.4 oxý 0&0, 01W 01%" 0VW aso os"'J' 0 00 00 *A. 0,31V 01'.0', ow"', 0ý0 %P& PAN jog" Test date

7-day IC25 = 25% inhibition concentration. An estimation of the concentration of potassium chloride that would cause a 25% reduction in Pimephales growth for the test population.

Central Tendency (mean IC25 )

Warning Limits (mean 1C.5 + SA.+/-0 or 5A.75) Graphs generated from associated excel spreadsheet.

......... Control Limits (mean IC + S S or 2 Standard Deviations) Excelspre"dsheetent dbV:J.Sumner 25 -A.25' A.90' Reviewed loy:

vi Kelley E.Keenan Initials: _ _"-

l*5 OETS.

Environmental Testing Solutions, Inc.

Pimephalespromelas Chronic Reference Toxicant Control Chart State and USEPA Laboratory Laboratory USEPA USEPA rD Test number Test date 7-day W, CT S Control Limits Sý Warning ULilts Control Lihmits SA.

SAs 7s Warning Limits SAAM Control Umits CV m (g/L KCI) (/L KCI) CT-2S CT+2S CT-S.uo CT+S, 4 , CT-SO. CT+S. CT- SLS CTO+S&7 CTr-SLI Cr + SA&.

K (D 1 12-07-10 0.77 0.72 0.04 0.64 0.80 0.09 0.64 0.81 0.15 0.88 0.27 0.57 0.45 1.00 0.33 0.40 1.05 0.06 2 01-18-11 0.78 0.73 0.04 0.64 0.81 0.09 0.64 0.82 0.15 0.57 0.88 0.28 0.45 1.00 0.33 0.40 1.06 0.06 3 01-25-11 0.70 0.73 0.04 0.64 0.81 0.09 0.64 0.81 0.15 0.57 0.88 0.28 0.45 1.00 0.33 0.40 1.05 0.06 4 02-08-11 0.69 0.72 0.04 0.64 0.81 0.09 0.64 0.81 0.15 0.57 0.87 0.27 0.45 1.00 0.33 0.40 1.05 0.06 5 02-15-11 0.68 0.72 0.04 0.64 0.80 0.09 0.64 0.81 0.15 0.57 0.88 0.27 0.45 1.00 0.33 0.40 1.05 0.06 6 03-08-11 0.71 0.72 0.04 0.64 0.80 0.09 0.64 0.81 0.15 0.57 0.87 0.27 0.45 1.00 0.33 0.40 1.05 0.06 7 03-08-11 0.70 0.72 0.04 0.64 0.80 0.09 0.63 0.81 0.15 0.57 0.87 0.27 0.45 0.99 0.32 0.40 1.04 0.05 8 03-29-11 0.75 0.72 0.04 0.64 0.80 0.09 0.63 0.81 0.15 0.57 0.87 0.27 0.45 0.99 0.32 0.40 1.04 0.05 9 03-31-11 0.75 0.72 0.04 0.64 0.80 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0.45 0.99 0.32 0.40 1.04 0.05 10 05-03-11 0.78 0.72 0.04 0.64 0.80 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0.45 0.99 0.32 0.40 1.04 0.05 11 06-07-11 0.76 0.72 0.04 0.64 0.80 0.09 0.63 0.81 0.15 0.57 0.87 0.27 0.45 0.99 0.32 0.40 1.04 0.05 12 06-14-11 0.63 0.71 0.04 0.63 0.80 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.99 0.32 0.39 1.04 0.06 13 07-12-11 0.67 0.71 0.04 0.63 0.80 0.09 0.63 0.80 0.15 0.56 0.87 0.27 0.44 0.99 0.32 0.39 1.04 0.06 14 07-19-11 0.63 0.71 0.05 0.62 0.80 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.06 15 08-09-11 0.67 0.71 0.05 0.62 0.80 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 16 09-13-11 0.66 0.71 0.05 0.61 0.80 0.08 0.62 0.79 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 17 10-04-11 0.73 0.71 0.05 0.61 0.81 0.09 0.62 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 18 10-11-11 0.76 0.71 0.05 0.62 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.99 0.32 0.39 1.03 0.07 19 10-18-11 0.67 0.71 0.05 0.61 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 20 11-08-11 0.79 0.71 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 Note: 7-d IC. = 7-day 25% inhibition concentration. An estimation of the concentration of potassium chloride that would cause a 25% reduction in Pimephales growth for the test population.

CT= Central tendency (mean IC,)..

S = Standard deviation of the IC2,values.

Laboratory Control and Warning Limits Laboratory control and warning limits were established using the standard deviation of the IC2svalues corresponding to the 10th and 25th percentile CVs. These ranges are more stringent than the control and warning limits recommended by USEPA for the test method and endpoint.

$a = Standard deviation corresponding to the 10t" percentile CV. 15,. = 0.12) 5* = Standard deviation corresponding to the 25P percentile CV. (Sý.as= 0.21)

USEPA Control and Warning Limits S,,,, = Standard deviation corresponding to the 7 5 thpercentile CV. (5.75 = 0.38)

S"= Standard deviation corresponding to the 90P percentile CV. (SAo= 0.45)

CV= Coefficient of variation of the IC values.

USEPAL 2000. Understanding and Accounting for Method VariabilityInWhole EffluentToxiity Applications under the National Pollutant DischargeElimination Program. EPA-833-R-O0-003.

USEnvironmental Protection Agency, Cncnnati, OH.

File: ppkclcr.110811Idsx Entered by: J. Sumner Reviewed by -

" 'II eTS Environrnental Testing Solutions, Inc.

Pimephalespromelas Chronic Reference Toxicant Control Chart Precision of Endpoint Measurements Organism Source: Aquatox, Inc.

1.25 I I I ' l . I I I I I I I I I I I I I I I

"" 1.00

- 0.75

.- E- 0.50 0 b 0.25 USEPA Acceptance Criteria (> 0.25 mg per surviving larvae)

- I I I I I I I I I I I I I i I I I I I -

I I I I I I I I I I I I I I I I I I I .

30 Kentucky Acceptance Limit (< 30.0%)

o.2 20 54_ 0 10 4)D 0 0 U

I i i I I I I I I I i I I I I i i I I I I I I I I I I I I I 30 ------

USEPA Upper PMSD Bound (90' percentile < 30.0%)

20 a.

10 0 0I I SI I I I I I I I I I I I I 0 I, I NN Test date

- Control Reproduction, Coefficient of Variation (CV), or Percent Minimum Significant Difference (PMSD) PMSD is the minimum significant difference between the control and treatment that can be declared statistically significant.

Central Tendency (mean Control Growth, CV, or PMSD)

......... Control Limits (mean Control Growth, CV, or PMSD +/- 2 Standard Deviations)

Graphs generated from associated excel spreadsheet.

"i'$n6ft9viY6,Kelley E.Keenan Excel spreadsheet ented by: J. Sumner Initials: Reviewed by:

Precision of Endpoint Measurements S Pimephalespromelas Environmental*esoudon, Inc Chronic Reference Toxicant Data Test date Control Control Control Mean Test number Survival Cr CV CT MSD PMSD Cr Growth for Control Growth for Control

() (m/larvae (mg/larvae) (%) for PMSD (%)

Growth CV (%)

1 12-07-10 100 1.080 8.3 0.15 13.9 2 01-18-11 97.5 0.806 0.943 5.8 7.0 0.09 11.0 12.4 3 01-25-11 100 0.608 0.831 4.6 6.2 0.08 13.2 12.7 4 02-08-11 100 0.817 0.828 2.6 5.3 0.06 7.8 11.5 5 02-15-11 100 0.701 0.802 3.3 4.9 0.12 16.8 12.5 6 03-08-11 100 0.953 0.827 3.1 4.6 0.08 8.4 11.9 7 03-08-11 100 1.006 0.853 4.7 4.6 0.13 12.7 12.0 8 03-29-11 100 0.829 0.850 6.3 4.8 0.09 10.3 11.8 9 03-31-11 100 0.675 0.830 3.2 4.6 0.09 12.8 11.9 10 05-03-11 100 0.867 0.834 6.3 4.8 0.08 8.8 11.6 11 06-07-11 97.5 0.839 0.834 5.8 4.9 0.11 12.7 11.7 12 06-14-11 100 0.758 0.828 14.5 5.7 0.13 16.8 12.1 13 07-12-11 97.5 0.872 0.831 6.0 5.7 0.09 9.8 11.9 14 07-19-11 100 0.839 0.832 5.4 5.7 0.06 7.2 11.6 15 08-09-11 100 0.662 0.821 6.6 5.8 0.07 10.6 11.5 16 09-13-11 100 0.848 0.822 6.7 5.8 0.12 13.6 11.6 17 10-04-11 100 0.768 0.819 16.8 6.5 0.13 16.9 12.0 18 10-11-11 100 0.681 0.812 4.3 6.3 0.07 9.9 11.8 19 10-18-11 100 0.693 0.805 4.8 6.3 0.07 10.0 11.7 20 11-08-11 100 0.792 0.805 10.3 6.5 0.12 15.6 11.9 Note: CV = Coefficient of variation for control growth.

Lower CV bound determined by USEPA (l 0 th percentile) = 3.S%.

Upper CV bound determined by USEPA ( 9 0 th percentile) = 20%

MSD = Minimum Significant Difference PMSD = Percent Minimum Significant Difference PMSD Is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Lower PMSD bound determined by USEPA (10' percentile) = 12%.

Upper PMSD bound determined by USEPA (90 percentile) = 30%.

CT = Central Tendancy (mean Control Growth, CV, or PMSD)

USEPA. 2000. Understanding and Accounting for Method Variability in Whole Effluent Toxicity Applications Under the National Pollutant Discharge Elimination Program.

EPA-833-R-00-003. US Environmental Protection Agency, Cincinnati, DH.

USEPA. 2001a, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes 1 and 2 Appendix. EPA-821-8-01-004 and EPA-821-B-01-005. US Environmental Protection Agency, Cincinnati, OH.

File: ppkclcr 11081 1.xlsx Fraae 7.91"oy KeeK0 Entered by: J.Su ner epend*7ve Kelley E. Keenan Reviewed by:

Initials:W

Ir ES SEnv ,ommn nnal Testig So talons,Inc.

Potassium Chloride Chronic Reference Toxicant Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephalespromelas PpKCICR Test Number: ZL HIM Dilutionpreparationinformation:

KCI Stock INSS number: lt4% 1000 50 g KCI/L:

Comments:

Stock preparation:

Dissolve 50 KCI in I-L Milli-Q water.

Dilution prep (mg/L) 450 600 750 900 1050 Stock volume (mL) 9 12 15 18 21 Diluent volume (mL) 991 988 985 982 979 Total volume (mL) 1000 1000 1000 1000 1000 Test organism information: Test information:.

Organism age: 2 .- IS j*jYt- 6Q Randomizing template: I bit Date and times organisms l.j\ I Incubator number and a were born between: shelf location:

Organism source: 'l * - Artemia CHM number: C/4W4S Drying informationfor weight determination:

Transfer vessel pH = 1.60 S.U. Date / Time in oven: \ IO information: 0 Temperature = "4,4 C Initial oven tentperature:

Average transfer volume: Date / Time out of oven: Il- I.' i1 Q2.iS O., To - Final oven temperature: 60, C.

Total drying time: 2*I16u, Dailyfeeding and renewal information:

Day Date Morning feeding Afternoon feeding Test initiation, renewal, MHSW or termination batch used Time o *i1-0,&~tI ---- , Analvyt Time

U Analyst O* Time

\:% Analyst O l0,t n 2_ Wadi-

'q-t*t II W06j¢0 >4

4 v-txv'oo *" ItS%

I2'cO *-.. il-o,4,- A tl-oq-tI g*

5 6 IIl'[ "11--,l l ~~ ' -s" I1"0. 11 7 I l I uSRA" t100 I

Control information: Acceptance criteria Summary of test endpoints:

% Mortality: 1. ! 20%

< 7-day LCso f 1.4, -

Average weight per initial larvae: / NOEC ('O Average weight per surviving larvae: 0.101 7., 0.25 mg/larvae LOEC "I ChV t(7.')a IC25 IllS.-

"I 10 Pange 80 of K SOP AT21 - Exhibit AT21.1, revision 06-01-11 E. Keenan initals:

Initepenae l revlewy Kelley V

Page 2 of 5 ET l1*

a~rnme~htIIl~ng Samo.utmInc Species: Pimephalespromelas PpKCICR Test Number: .2A1 Survival and Growth Data Day Control 450 mg KCI/L 600 mg KCI/L A B C D E F G H I J K L 0 I 1( 10]

10 /0 t 0 to /0 /0 /0 10 /0 /0 /0 /010fl 0I /l /0 /0 to 1

2 i0 / 0 /0 /0 I(0 /I(0 /0C 0 'o0 '0 '0 3 0 0 60/0 U0 to 10/0(0 /0 I0 4 /0 0010 / /0 /D0 1) / i 100

/1 6 1 to t0 t o1o b, /0' 1 o 1 t0 o 9o 7

(0 /0 A=Paneightmg)o10 C)0 0 II0 60 00 /Q t9 A yPan weight (m:g)

Tray color code:: Dal .&I.. ..

Analyst: ___ 9_________ 1~. 4.L1A 1.44 It.S4 l&'$9 v6. Se, IS.4. IUz4 M..1& 14.0o i6.t.4 Date: __.____ *___ _I B = Pan + Larvae weight (mg)

Analyst: M4____________

cC Date: V *, Ut.L t ti. ZI.S 4 U.OO L,.V&. to..5 M. i4 1. M .. to ,*1.2t C = Larvae weight (mg) = B - A Hand calculated. '".0c7.* 1 / *e .4.'14L . a rL

,'S ".1 o.0t.do 6 , . o3i S.65 Analyst:

I Weight per initial number of larvae (mg)

= C / Initial number of larvae A

Hand calculated. 'I Analyst: 0 0D 0*

Average weight per initial Percent number of larvae (mg) reduction from control 0."t2' 0. 8b -, q 7. 0.68 t ,.o7.

M%

Comment codes: c = clear, d = dead, fg = fungus, k = killed, m = missing, sk = sick, sm = unusually small, Ig = unusually large, d&r = decanted and returned, w = wounded.

Comments:

SOP AT21 - Exhibit AT21.1, revision 06-01-11 PPaee e81 Inpender ~le~w y KflO Kelley E E. Keenan Initials: i7W

S Page 3 of 5 0

0ETS EnvhnmantalTestln 9 In,.

Species: Pimephalespromelas PpKCICR Test Number: __A I

I I

I A = Pan weight (nmg)

Tray color code:: W.r_-

Analyst:

Date: 1o-iAG .

1.11It- I1 '.S I 411] 4.,$ ,s.C* l4.,t* 14 .'st I*.

%ýA 1 .,'s 1,5.94 B = Pan + Larvae weight (nag)

A nalyst: ____1 4___1 .__1 4__1_,____

Date: 11-_1_11_ _ __ Iq.si* lq.4' I___ tt.* . .'Lq l6.4i . :4.1c I4.,* 14.4 C = Larvae weight (mg) = B - A Handecalculated. {,(0 . -.b6 ($.UlJ 6.O0 -. D#,, 3,44 7.t 2.AS 1.-11 O.PH" 0.MO Analyst:

Weight per initial number of larvae (mg)

= C /Initial number of larvae Hand calculated.

Analyst: *Y r"

1W 0, Average weight per initial Percent number of larvae (mg) reduction from control Comment codes: c = clear, d = dead, fg = fungus, k = killed, m = missing, sk = sick, sm = unusually small, Ig= unusually large, d&r = decanted and returned, w = wounded.

Comments:

SOP AT21 - Exhibit AT21.1, revision 06-01-I1 naKelley E.Keenan Initials:

I . t . ... . . .

ETS 0

Pimephalespromelas Chronic Reference Toxicant Test EPA-821-R-02-013, Method 1000.0 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Test nmnben 241 (Environmental Testing Solutions, Inc.

Test dates: November 08-15, 2011 Confi(nWL KiI] RKt Hs hnhlida lberof Rnennml baof A-P-namMl.tnsa) n-*P., L. ta.aaetiniaj- "aW/iS*I nln M-ma)5.iht lnln Coefldimt uvdlittan W.lIedb/1nidt1nuanIraI amimihslfNMwn 5htIhdstiACntildaJWt*oi m P t r-dsdlnfoms hAnS. wd*5*snis A-B o-beraofhlase.(.W5 nansamof enn. (5.61 ben..

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A 10 10 14.02 21.61 7.59 0.759 0.759 control B 10 10 14.65 21.72 7.07 0.707 0.792 103 0.707 100.0 0.792 103 Not applicable C 10 10 14.44 22.46 8.02 0.802 0.802 D 10 10 12.85 21.64 8.99 0.899 0.899 E 10 10 13.39 21-56 8.17 0.817 0.817 F 10 10 13.58 22.00 8.42 0.842 083 6.2 0.842 100.0 0.803 6.2 -1A

- 10 10 13.55 21.77 8.22 0.822 0.822 H 10 10 13A45 20.75 7.30 0.730 0.730 I 10 10 13.24 20.14 6.90 0.690 0.690 600 1 10 10 12.76 19.36 6.60 0.660 0.697 11.3 0.660 97.5 0.681 14.8 14.0 K 10 10 14.01 22.10 8.09 0.809 0.809 L 10 9 13.64 19.29 5.65 0.628 0.565 M 10 8 1439 21.28 6.89 0.861 0.689 750 N 10 9 12.93 19.35 642 0.713 0.8 83 0.642 82. 0.681 4.0 16.5 0 10 8 13.05 19.11 6.36 0.795 0.636 P 10 8 14.82 21.60 6.78 0.848 0.678 Q 10 4 14.28 17.28 3.00 0.750 0.300 900 R 10 5 14.30 17.49 3.19 0638 0.734 131 0.319 40.0 0.292 20.5 63.1

$ 10 3 15.95 18.01 2.06 0.687 0.206 T 10 4 14.99 18A3 3.44 0.860 0.344 U 10 3 14.51 16.96 2.49 0.817 0.245 1050 V 10 2 13:A9 14.70 1.21 0.605 18.3 0.12117.5 . 8.

W 10 1 13.13 14.07 0.94 0.940 0.094 X 10 1 13.84 14.74 0.90 0.900 0.090 Dunnett's MSO value: 0.1234 MSD = Minimum Significant Difference PMISD: 15.6 PMSD = Percent Minimum Significant Difference PMSD is a measure of test precision. The PMSD Is the minimum percent difference between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Lower PIYSD bound'determined by USEPA (10th percentile) = 12%.

Upper PMSD bound determined by USEPA (90th percentile) = 30%.

Lower and upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET Interlaboratory Variability Study (USEPA, 2001a; USEPA, 2001b).

USEPA. 2001a, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes I and 2-Appendix. EPA-821-B-01-004 and EPA-821-B-01-005. US Environmental Protection Agency. Cincinnati, OH.

File: ppkclcr 1081I.xlsx Enteed by: J SnAn Reviewed h _*

Statistical Analyses 1ý Envi r - meta [T emtng Solutions,Inc .

Larval Fish Growth and Survival Test-7 Day Survival Start Date: 111812011 Test ID: PpKCICR Sample ID: -REF-Ref Toxicant End Date: 11/15/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: KCL-Potassium chloride Sample Date: Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelas Comments:

Conc-mg/L . 1 2 3 4 D-Control 1.0000 1.0000 1.0000 1.0000 450 1.0000 1.0000 1.0000 1.0000 800 1.0000 1.0000 1.0000 0.9000 750 0.8000 0.9000 0.8000 0.8000 900 0.4000 0.5000 0.3000 0.4000 1050 0.3000 0.2000 0.1000 0.1000 Transform: Arcsin Square Root Rank 1-Tailed Number Total Conc-mg/L Mean N-Mean Mean Min Max CV% N Sum Critical Resp Number D-Control 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4 0 40 450 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4 18.00 10.00 0 40 600 0.9750 0.9750 1.3713 1.2490 1.4120 5.942 4 16.00 10.00 1 40

750 0.8250 0.8250 1.1426 1.1071 1.2490 6.209 4 10.00 10.00 7 40

900 0.4000 0.4000 0.6836 0.5796 0.7854 12.289 4 10.00 10.00 24 40 "1050 0.1750 0.17590 0.4217 0.3218 0.5796 29.582 4 10.00 10.00 33 40 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.9221 0.884 0,22125 0.55458 Equality of variance cannot be confirmed Hypothesis Test (1-tall, 0.05) NOEC LOEC ChV TU Steels Many-One Rank Test 600 750 670.82 -

Treatments vs D-Control

- Maximum Likelihood-Probit Parameter Value SE 95% FiducIal Limits Control ChI-Sq Critical P-value Mu Sigma Iter Slope 12.5168 1.72316 9.13938 15.8942 0 0.53062 7.81472 0.91211 2.94187 0.07989 3 Intercept -31.823 5.06011 -41.741 -21.905 TSCR 1.0 Point Probits mglL 95% Fiducial Limits 2.674 570.178 485.278 827.787 0.9 ECOI EC05 3.355 646,334 574.314 695.18 0.8 ECIO 3.718 891.006 627.535 734.882 0.7 EC15 3.964 722.878 865.625 763.604 EC20 4.158 749.254 696.999 787.84

0.6 EC25 4.326 772.648 724.528 809.874 0o0.5 EC40 4.747 834.888 794.89 872.454 EC50 5.000 874.719 836.514 916.719 8 04 EC60 5.253 916.451 876.887 986.998 0.3 EC75 5.674 990.275 942.018 1063.86 0.2 EC80 5.842 1021.19 967.704 1106.64 EC85 6.036 1058.45 997.84 1159.47 0.1 EC90 8.282 1107.27 1036.32 1230.47 0.0 EC95 6.645 1183.81 1095 1345.11 1 10 100 1000 10000 EC99 7.326 1341.92 1212.04 1592.58 Dose mg/L Dose-Response Plot 0.9 0.8 0.7 0.8 0.5 C 0.4 0.3 0.2 0.1 File: ppkclcr 11081l.xlsx P.Initials:en n en4tof1 rview bV Kelley E. Keenan Entered by: J. Sunner Reviewed by:

Initials: -

-TS SET Statistical Analyses

~arvai I-ISn Urowtn and Survival Test-i Day G~0Wtfl Larval F-ish Growth and Survival Test-7 Day Growth Start Date: 11/8/2011 Test ID: PpKCICR Sample ID: REF-Ref Toxicant End Date: 11/15/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: KCL-Potassium chloride Sample Date: Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Plmephales promelas Comments:

Conc-mglL 1 2 3 4 D-Control 0.7590 0.7070 0.8020 0.8990 450 0.8170 0.8420 0.8220 0.7300 600 0.6900 0.6600 0.8090 0.5650 750 0.6890 0.6420 0.6360 0.6780 900 0.3000 0.3190 0.2060 0.3440 1050 0.2450 0.1210 0.0940 0.0900 Transform: Untransformed 1-Tailed Isotonic Conc-mgIL Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Mean N-Mean D-Control 0.7918 1.0000 0.7918 0.7070 0.8990 10.277 4 0.7973 1.0000 450 0.8028 1.0139 0.8028 0.7300 0.8420 6.190 4 -0.194 2.160 0.1234 0.7973 1.0000 600 0.6810 0.8601 0.6810 0.5650 0.8090 14.773 4 1.957 2.180 0.1234 0.6810 0.8542 750 0.6613 0.8352 0.6613 0.6360 0.6890 3.962 4 0.6613 0.8294 900 0.2923 0.3691 0.2923 0.2080 0.3440 20.818 4 0.2923 0.3666 1050 0.1375 0.1737 0.1375 0.0900 0.2450 53.074 4 0.1375 0.1725 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.9603 0.805 0.2313 -0.214 Bartletts Test Indicates equal variances (p = 0.55) 1.20701 9.21035 Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnett'sTest 600 >600 0.12335 0.1558 0.01814 0.0064 0.11111 2,9 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point mg/L SD 95% CL(Exp) Skew IC05 501.44 73.80 130.99 683.88 -2.1748 IC16 552.87 59.38 410.36 758.62 -0.1688 IC15 625.35 80.46 443.90 833.78 0.0697 1.0 C20 759.53 65.63 433.55 777.98 -1.9095 0.9 1 rIC25 775.74 29.52 682.07 793.43 -6.36901 IC40 824.35 9.16 789.14 844.18 -0.5613 0.8 IC50 856.76 9.67 825.72 880.40 -0.2566 0.7-C 0.6-05-0.4:

0.3-0.2-

/

0.1 -

0.0

-0.1 0 500 1000 1500 Dose mg/IL Dose-Response Plot 1-tail, 0.05 level of significance File: ppkclcr I 1051I.Ae Entered by:J. Sumner PiNInniatln efiOB~y Kelley E. Keenan Reviewed by:*

Initials:

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Environmental Testing Solutions, Inc.

Pimepholespromelas Chronic Reference Toxicant Test Control Limits Determination (D.

rn State and USEPA Laboratory Laboratory USEPA USEPA Test number Test date 7-day ICa CT S Control Unmits S&.o Warning ULmits So2 Control Limits S&75 Warning mits S,&w Control Umits CV (g/L KCI) (g/L KCI) CT-2S CT+2S CTa-SA.O CT + S&. CT-Sn, Ca.s CT-S 75 T + S. C- So CT÷+ 5Aso 1 12-07-10 0.77 2 01-18-11 0.78 0.78 0.01 0.75 0.80 0.09 0.68 0.87 0.16 0.61 0.94 0.29 0.48 1.07 0.35 0.43 1.13 0.02 3 01-25-11 0.70 0.75 0.04 0.66 0.84 0.09 0.66 0.84 0.16 0.59 0.91 0.29 0.47 1.04 0.34 0.41 1.09 0.06 4 02-08-11 0.69 0.74 0.05 0.64 0.83 0.09 0.65 0.82 0.15 0.58 0.89 0.28 0.46 1.02 0.33 0.40 1.07 0.06 5 02-15-11 0.68 0.72 0.05 0.63 0.82 0.09 0.64 0.81 0.15 0.57 0.88 0.28 0.45 1.00 0.33 0.40 1.05 0.07 6 03-08-11 0.71 0.72 0.04 0.63 0.81 0.09 0.64 0.81 0.15 0.57 0.87 0.27 0.45 1.00 0.32 0.40 1.05 0.06 7 03-08-11 0.70 0.72 0.04 0.64 0.80 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0.45 0.99 0.32 0.40 1.04 0.06 8 03-29-11 0.75 0.72 0.04 0.64 0.80 0.09 0.64 0.81 0.15 0-57 0.87 0.27 0.45 1.00 0.32 0.40 1.05 0.06 9 03-31-11 0.75 0.73 0.04 0.65 0.80 0.09 0.64 0.81 0.15 0.57 0.88 0.28 0.45 1.00 0.33 0.40 1.05 0.05 10 05-03-11 0.78 0.73 0.04 0.65 0.81 0.09 0.64 0.82 0.15 0.58 0.88 0.28 0.45 1.01 0.33 0.40 1.06 0.06 11 06-07-11 0.76 0.73 0.04 0.65 0.81 0.09 0.65 0.82 0.15 0.58 0.89 0.28 0.46 1.01 0.33 0.40 1.06 0.05 12 06-14-11 0.63 0.73 0.05 0.63 0.82 0.09 0.64 0.81 0.15 0.57 0.88 0.28 0.45 1.00 0.33 0.40 1.05 0.07 13 07-12-11 0.67 0.72 0.05 0.62 0.82 0.09 0.63 0.81 0.15 0.57 0.87 0.27 0.45 0.99 0.32 0.40 1.05 0.07 14 07-19-11 0.63 0.71 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.99 0.32 0.39 1.04 0.07 15 08-09-11 0.67 0.71 0.05 0.61 0.82 0.09 0.63 0.80

0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 16 09-13-11 0.66 0.71 0.05 0.60 0.81 0.08 0.62 0.79 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 17 10-04-11 0.73 0.71 0.05 0.61 0.81 0.09 0.62 0.79 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 18 10-11-11 0.76 0.71 0.05 0.61 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 19 10-18-11 0.67 0.71 0.05 0.61 0.81 0.09 0.62 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 20 11-03-11 0.78 0.71 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.98 0.32 0.39 1.03 0.07 Note: 7-d ICs = 7-day 25% Inhibition concentration. An estimation of the concentration of potassium chloride that would cause a 25% reduction in Pimephales growth for the test population.

CT = Central tendency (mean IC55).

S = Standard deviation of the IC,, values.

Laboratory Control and Warning Limits Laboratory control and warning limits were established using the standard deviation of the IC, values corresponding to the 10th and 25th percentile CVs. These ranges are more stringent than the control and warning limits recommended by USEPA for the test method and endpoint.

S*s = Standard deviation corresponding to the lt'h percentile CV. (Saja = 0.12)

S,,, = Standard deviation corresponding to the 2 SP percentile CV. (SA, = 0.21)

USEPA Control and Warning Limits S7o= Standard deviation corresponding to the 75m percentile CV.(SA,* = 0.38)

S_.= Standard deviation corresponding to the 9 0 ' percentile CV. (Sane = 0.45)

CV= Coefficient of variation of the IC25svalues.

USEPA. 2000. undem~tandingand Accounting for Method VariabilityinWhole EffluentTodicity Applications Under the National Poflutant DischargeElimination Program. EPA-833-R-O-003.

US Environmental Protection Agency, Cncinnati, OH.

Fie: ppk*crit083i.xlsx

,amed by.J St Renewed by-.C

Page 4 of 5 ETS

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Species: Pimephales promelas PpKCICR Test Number: __7_q Daily Chemistry: ________________________

I- Day I Analyst Concentration IParameter I pH (S.U.)

DO (mc/L)

*.5 - av Conductivity (jimhos/cm)

CONTROL Alkalinity (mg CaCO3 /L)

Hardness (mg CaCO 3/L)

Temperature (0c)

DO PH (sg/.)

(SU. 1;lp

___a . 7 **

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190 ~o 450 mg KCIIL Conductivity (4*mhos/cm) 1100 I0q0 1090 Temperature - ,. W 600 mg KCI/L Conductivity 1 0 (gimhos/cm) )30 I5qO Temperature - -:i & "

(0C)

DO (mg/L) ' d ' ' °' 1 750 g Kl/LConductivity 1100119OO O 750KCILm(nhos/cm) mg Temperature -q . I, 2q,1 2. 2 "V. V4.q pH (S.U.) I,',1i',"aic 13'.14,1(0*.)*

, 3 900 Mg KCIIL Conductivity Temperature 22q .' "

i.q. 0 j..,

(°C) pH (S.U.)

DO (mg/L)

Conductivity 1050 mg KCI/L

([tmhos/cm) 2-1000 Temperature

(°C' STOCK Conductivity

_(jmhos/cm) I Initial Final Initial 11 Final I Initial j SOP AT21 - Exhibit AT2 1.1, revision 06-01-11 PaI'ge B7 of 10 E pen ri 1My Kelley E.Keenan Initials:

lETS Page 5 of 5 Eqr41Emsman TibtingSolution n.c Species: PimephalesPromelas PpKCICR Test Number: ZAI Day 3 4 5 6

__ __ _Analyst ALA MA IM11(L I v 111O T 1 F Concentration Parameter Conductivity (gmhos/cm) 30( 3O0 CONTROL Alkalinity 11.

(1111 CaCO 3/L)

Hardness \

(mg CaCO 3/L) ' 0; Temperature WA

_____ _____ (OC) pH (S.U.) 0.o5 .o el .00 .Q, A Conductivity IOOO 450 ing KCIIL (ji00 m 10 Temperature C(C)

C -usA t,.'4 2{q. 4, . ,4Q ..

. pH (S.U.) 61,03 {,* ¢I ),O eA", f*.f

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600OgKCI/L 1(00 1+ 'ITo-to Conductivity (iimhoslcm)

Temperature 600 mng KCI/L 0

(( 0)mo/cm) 13'0O 5"40 1*O M. at Q~

pH (S.U.) 0,0V q3/t S.ot "___,_

750 mg KCI/L Conductivity (pimhos/cm) 15l Temperature "40 , 24.S* "j..6 (0c) 1...__ 2.

pH (S.U.) 10.I _ __ .* . 4 j j13 .01 -

900 mg KCL onductivity (gnihos/cm) 100 90(3 TO o) ** ". .*

DO (mg/L> ,44*

Temperature , . . -, , .

_pH (S.U.) 10.1 ..4 t DO (mg/L).0 (Oc) 1050 mg KCI/L Conductivity AoD'lI Temperature zq~i~q.~ zq10 i. 214. 9 {

T- - - ,- - - -

Initial m-InitialIm - nitial .inm Final Initial Final i -

Initial ~

Final SOP AT21 -Exhibit AT21.1, revision 06-01-11 "IM n 'd*)'-vi.q; Kelley E. Keenan Initials:

-Ceriodaphnia dubia

Chronic Reference Toxicant Control Chart Environmental Testing Solutions, Inc.

I I I I I I I I I I I I d I I eIv I IS I I 1.14 USEPA ControlLimits (+/-2 StandardDeviations) 1.12 -

1.10 ............ ............... ........................................ .....

1.08 . ..................

1.06 1.04 1.02 I I I

I I I i

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2.5 I I I I ! I I l I I I I I I ! I f I I USEPA Warning and ControlLimits ( 7 5 th and 9 0 th Percentile CVs)

U 2.0 1.5 0 9e U' 1.0 0.5 ................................................................. e..... e ...........

r--

I I I I I I I I I I I I I I I i I I I I 1.4 I I I I I I I I I I I I I I I I I I I I Laboratory Warning and Control Limits (10th and 25th PercentileCVs) 1.3

. .................................................... a...................

1.2 1.1 1.0 0.9 ................................. a.....C....*........ee.. ... e..........

I I I I I I I I I I f I I I I I I 0.8 Test date

-- 7-day IC 2s = 25% inhibition concentration. An estimation of the concentration of sodium chloride that would cause a 25% reduction in Ceriodaphniareproduction for the test population.

Central Tendency (mean IC 2 5)

Warning Limits (mean IC 25 +/-SA-10 or SA75)

......... Control Limits (mean IC 25 +/- Sk25' SA.90, or 2 Standard Deviations)

Graphs generated from associated excel spreadsheet.

Page 89 of 102 Excelspreadsheet ered by: J. Sumner Independent review by Kelley E. Keenan Reviewed by: 7 initials:P,

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"ETS l,'Ceriod aphnia dubia r

io

-X Chronic Reference Toxicant Control Chart M. ( Environmental Testing Solutions, Inc.

m State and USEPA Laboratory Laboratory USEPA USEPA a Test number Test date 7-day ICa CT s Control Umits S&, Warning UmLits Ss Control Limits S. Warning Limits SAM- Control Limits CV (g/L Naci) (g/L NaCI) CT-2S CT+zS CTa-S.. CT + a'.

CT-S. CT+Sý CT- Sk7. CT + S,7 . CT-Sm CTa+S 1 07-13-10 1.07 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.74 0.01 2 08-03-10 1.06 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.74 0.01 3 09-14-10 1.07 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 4 10-05-10 1.08 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 5 10-05-10 1.08 1.07 0.01 1.05 1.10 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.74 0.01 6 11-02-10 1.09 1.07 0.01 1.05 1.10 0.09 0.99 1.16 0.18 0.89 1.26 0.48 0.59 1.56 0.67 0.41 1.74 0.01 7 12-07-10 1.06 1.07 0.01 1.05 1.10 0.09 0.99 1.16 0.18 0.89 1.26 0.48 0.59 1.56 0.67 0.41 1.74 0.01 8 01-18-11 1.06 1.07 0.01 1.05 1.10 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.56 0.66 0.41 1.74 0.01 9 02-08-11 1.06 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.74 0.01 10 03-08-11 1.05 1.07 0.01 1.05 1.10 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.74 0.01 11 04-05-11 1.06 1.07 0.01 1.05 1.09 0.09 0.98 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 12 04-05-11 1.07 1.07 0.01 1.05 1.09 0.09 0.98 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 13 05-03-11 1.07 1.07 0.01 1.05 1.09 0.09 0.98 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 OA1 1.73 0.01 14 06-07-11 1.05 1.07 0.01 1.05 1.09 0.09 0.98 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 15 07-12-11 1.06 1.07 0.01 1.04 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 16 08-09-11 1.06 1.07 0.01 1.04 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 .0.66 0.41 1.73 0.01 17 09-13-11 1.06 1.07 0.01 1.04 , 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 18 10-04-11 1.07 1.07 0.01 1.04 , 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 19 10-11-11 1.06 1.07 0.01 1.05 1.08 0.09 0.98 1.15 0.18 0.88 1.25 0.48 0.59 1.55 0.66 0.40 1.73 0.01 20 11-08-11 1.05 1.07 0.01 1.05 1.08 0.09 0.98 1.15 0.18 0.88 1.25 0.48 0.59 1.54 0.66 0.40 1.73 0.01 Note: 7-d IC2, : 7-day 25% inhibition concentration. An estimation of the concentration of sodium chloride that would cause a 25% reduction in Ceriodaphnia reproduction for the test population.

CT = Central tendency (mean IC2s).

S= Standard deviation of the IC., values.

Laboratory Control and Warning Limits Laboratory control and warning limits were established using the standard deviation of the IC25values corresponding to the 10th and 25th percentile CVs. These ranges are more stringent than the control and warning limits recommended by USEPA for the test method and endpoint.

Siu, = Standard deviation corresponding to the et? percentile CV. (SAID= 0.08) 5* = Standard deviation corresponding to the 25 percentile CV. (S,,5 = 0.17)

USEPA Control and Warning Limits

$,. = Standard deviation corresponding to the 75" percentile CV. (S,7s = 0.45)

S,.= Standard deviation corresponding to the 90e percentile CV. (S.,o = 0.62)

CV = Coefficient of variation of the IC25values.

USEPA. 2000. Understanding and Accounting for Method Variability in Whole Effluent Toxicity Applications Under the National Pollutant Discharge Elimination Program. EPA-833-R-00-003. US Environmental Protection Agency, Cincinnati, OH.

File: CdNaCICR 110811.xAsx Entered by'. J..Wpmner Reviewed by&__

Ceriodaphniadubia 0-ETS Chronic Reference Toxicant Control Chart 5 Environmental Testing Solutions, Inc. Precision of Endpoint Measurements 40

I I I I I II I I I I I I I I I I 0 35 - . ... .. ... . .. .. . . . . .. . .. .. .... .. . . . . .. . .. ... .. .

30 0

Lu 25 T 20 Lu 0

U 15 Minimum Acceptance Criteria (> 15.0 offspring per surviving female)

I i I i i i i I i i i i i i I I I I I 40 North Carolina Acceptance Limit (< 40.0%)

'~.2

.* ! 30 Kentucky Acceptance Limit (< 30.0%)

. 20 10 * . .......................................

50 I I I I .I I I I I I I I I I I USEPA Upper PMSD Bound (90'h percentile < 47.0%)

40 30 20 10 S. ................ . - I I I I .. ..... I I I I n

V Test date

Control Reproduction, Coefficient'of Variation (CV), or Percent Minimum Significant Difference (PMSD) PMSD is the minimum significant difference between the control and treatment that can be declared statistically significant.

-- -- Central Tendency (mean Control Reproduction, CV, or PMSD)

......... Control Limits (mean Control Reproduction, CV, or PMSD +/-2 Standard Deviations)

Graphs generated from associated excel spreadsheet.

Pa oe.91 of enlendent 102by Kelley E. Keenan evew Excel spreadsheet entered by: J. Sumner Initials: Pr Reviewed by: N

Precision of Endpoint Measurements

.ETS Environmental Testing Solutions, Inc.

Ceriodaphniadubia Chronic Reference Toxicant Data Test Test date Control Control Mean number cr CV CT MSD PMSD CT Survival Reproduction for Control Mean for Control

(%) (offspring/female) Reproduction (%) Reproduction (%) for PMSD (%)

(offspring/female) CV (%)

1 07-13-10 100 31.6 6.6 2.4 7.6 2 08-03-10 100 28.2 29.9 .4.8 5.7 2.3 8.0 7.8 3 09-14-10 100 32.9 30.9 4.7 5.3 1.8 5.3 7.0 4 10-05-10 100 30.2 30.7 8.7 6.2 3.0 10.0 737 5 10-05-10 100 30.8 30.7 7.6 6.5 2.0 6.5 7.5 6 11-02-10 100 30.5 30.7 4.0 6.0 2.2 7.3 7.5 7 12-07-10 100 32.2 30.9 4.9 5.9 2.0 6.2 7.3 8 01-18-11 100 31.9 31.0 4.8 5.8 2.0 6.3 7.2 9 .02-08-11 100 30.7 31.0 6.3 5.8 2.3 7.4 7.2 10 03-08-11 100 32.6 31.2 6.9 5.9 1.8 5.4 7.0 i1 04-05-11 100 32.8 31.3 4.8 5.8 2.0 6.0 6.9 12 04-05-11 100 33.0 31.5 5.9 5.8 2.2 6.7 6.9 13 05-03-11 100 30.9 31.4 4.5 5.7 1.9 6.1 6.8 14 06-07-11 100 28.9 31.2 .6.7 5.8 2.5 8.7 7.0 15 07-12-11 100 32.2 31.3 6.0 5.8 2.0 6.2 6.9 16 08-09-11 100 28.3 31.1 5.0 5.8 1.9 6.8 6.9 17 09-13-11 100 32.1 31.2 8.2 5.9 1.9 5.8 6.8 18 10-04-11 100 32.8 31.3 6.1 5.9 2.1 6.5 6.8 19 10-11-11 100 31.3 31.3 3.0 5.8 2.1 6.6 6.8 20 11-08-11 100 29.0 31.1 7.6 5.9 1.8 6.1 6.8 Note: CV = Coefficient of variation for control reproduction.

th Lower CV bound determined by USEPA (101 percentile) = 8.9%.

Upper CV bound determined by USEPA (90t percentile) = 42%

MSD = Minimum Significant Difference PMSD = Percent Minimum Significant Difference PMSD is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Lower PMSD bound determined by USEPA (l 0 th percentile) =13%.

Upper PMSD bound determined by USEPA ( 9 0 th percentile) = 47%.

CT = Central Tendancy (Mean Control Reproduction, CV, or PMSD)

USEPA. 2000. Understanding and Accounting for Method Variability in Whole Effluent Toxicity Applications Under the National Pollutant Discharge Elimination Program. EPA-833-R-00-003. US Environmental Protection Agency, Cincinnati, OH.

USEPA. 2001a, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes 1 and 2-Appendix. EPA-821-B-01-004 and EPA-821-B-01-005. US Environmental Protection Agency, Cincinnati, OH.

File: CdNaCICR_11081 .xlsx Entered by: J. Sugner P.AQ e2tof1enntreiew y Kelley E.Keenan Reviewed by:

Initials:

S Page 1 of 6 6

00*ETS Sodium Chloride Chronic Reference Toxicant Test (EPA-821-R-02-013 Method 1002.0)

Species: Ceriodaphniadubia CdNaCICR #: t2?'l Dilutionpreparationinformation: Comments:

NaCI Stock INSS number: IW Irii.s Stock preparation: 100 g NaCIIL:

Dissolve 50 g NaCI in 500 mL Milli-Q water.

Dilution prep (mr/L) 600 800 1000 1200 1400 Stock volume (mL) 9 12 15 18 21 Diluent volume (mL) 1491 1488 1485 1482 1479 Total volume (mL) 1500 1 1500 1500 1500 1500 Test organism source information: Test information:

Organism age: < 24-hours old Randomizing template color:

Date and times organisms were born wo--t\ cri "o To O 2. Incubator number and shelf between: location: 7-31 Culture board: - 1 A.-I Replicate number: 1 2 3 4 5 6 7 1 9 10,/ WT batch:

Culture board cup number: b 6 110 It I12 S t U "1.UIVA _.__

Transfer vessel information: H S,. S.U. Temperature = S-.O°C Selenastrum batch: "

Average transfer volume (mL): A. 0'4%5 ---

Daily renewal information:

Day Date Test initiation and feeding, MHSW Analyst renewal and feeding, or batch used termination time o i\-o'.i\i oOrx: '1-o4-i*IX

5 6 I11-1-,-%-!rI clb

o. ItO--tl'0 Controlinformation: Acceptance criteria Summary of test endpoints:

%of Male Adults: 07. < 20% 7-day LC50 )jqOo

% Adults having 3d Broods: 1 M7. 80% NOEC too

%Mortality: Q7. 20% LOEC 1000 Mean Offspring/Female: . -, > 15.0 offspring/female ChV aigA F-/jCV:" < 40.0 % ICs, Pew. of 102 SOP AT14 - Exhibit AT14.1, revision 06-01-11

-In oenae*tjevlew By Kelley E.Keenan Initials: =

J. Page 2 of 6 aETS Species: Ceriodaphniadubia CdNaCICR #: 12t1 CONTROL Survival and ReproductionData Replicate number Day 1 2 3 4 5 6 7 8 9 10 I Young produced O 0) 0 i0 0 0UO 0 1 '

2 Adult mortality Young produced Adult mortality JL C) 60

%. '% HI + I 1II_ ,*_T[

'. =

3 Young produced Adult mortality L

__0 a I'-

c5T0~2 lL- '1 '- 1 4 Young produced~ 5 1L i --- f-+/-TA -L Adult mortality ' - I'--" '--I 'I--I-_ I --

5 Young produced [ [ J I I IJ I: __

6oa 6 Adult mortality Adult mortaity produced Young produced TYoung I

[

0.. .

C.)"I I_ __

I

\.

" I C) '

I***

C i 0_)0 k

\.

01 t1'-

'k-2.C) 0 U 1 10

\-

'20 2

I "-_

- t._

't__.

c.)

To]a young produced 2A zJ a- -Z -4 LI Final Adult Mortality L -C- =_ C C. :- L Z. '.

Tafor 3ouBroods - 4 y 7 t".ý -_ '? *> *- Z*

Note: Adult mortality (L = live, D = dead), SB = split brood (single brood split between two days), CO = carry over (offspring carried over with adult during transfer).

Concentrationt

% Mortality: 0.W Mean.Offsprinemale: ".,0 600 mg NaCiJL Survival and ReproductionData Replicate number Day 1 2 3 4 5 6 7 8 9 10 I Young produced C) p p p p Adult mortality_ _. __

__]1J _ L

' - _L--

A d u lt m o rta lity __

0) Q 0 L_ '- _

-1 6 3 Young produced Adult mortality I 01c1 L II 3

5

, Young produced Adult mortality Adult mortality Young produced I

0%

T I Q_

%A to...-

~

_ %.-I LA 1 C jL C 1%_

12 I11 I _-

Ij~

C1I 1+

It.

6 6 You. prdcd0 Adl mortality Young produce Vi_~ 'L- __

1Q

'-IL 0

i (2)

'- 'H -

o -

I0LL

ý.I . *.(

Total young produced j 3o __ .3 -n 3'L 'Z 'A z Final Adult Mortality % I. I \.- \ý I_

ANote: Adult mortality (L = live, D = dead), SB = split brood (single brood split between two days), CO = carry over (offspring carried over with adult during transfer).

Concentration: '

%Mortality:

Mean Offspring/Female: A.2-I% Reduction from Control: 0-.617. 1 Page 94 of 102 SOP AT14 - Exhibit AT14.1, revision 06-01-11 Ind-epen gt review by Kelley E. Keenan Initials____

Page 3 of 6

ETS Species: Ceriodaphniadubia CdNaCICR #: 171 800 mg NaCI/L Survival and ReproductionData Replicate number Day 1 2 1 3 4 5 6 7 8 9 10 1 Young produced 0 1..C..... 1_.*..0 0 0.0 C..

0

___Adult Adult mortality mortalit t__ %,- I \,- I_ I_ \ - . - ,_- ".- V-2 Young produced 1C1I .. I C)~ ).oQ.e..o Adult motlty L-I L- I-- I_ -_ \- _ ___ %__ _

3 Young prod uced 0 0 CT0 cc)

Adult mortality _I %.- _*'i- '- I, J'--L _---

4 5

__IAdultYoung produced mortality Young produe I

toIJII.

\*,

%j 2.I I..

I' l

___~

~..to in.1i ..

N~ 1 %A_

I,*.-

61 Adult mortality Young produced C) ' V_. 1 0101

]___01010 Adult mortality 7 Young produced - - - %4 - 1 - -

Total young produced -3 12A n z2. 30 7_jk Final Adult Mortality J' - -

Note: Adult mortality (L = live, D = dead), SB = split brood (single broqd split betwedn two days), CO = carry over (offspring

-- I '- ..

carried over with adult during transfer).

Concentration:

%Mortalit: 0".

Mean Offspring/Female: U. 01,

% Reduction from Control: o .'S 7.

1000 mg NaCI/L Survival andReproductionData Replicate number Day _ 1 2 3 4 5 6 7 1 8 9 10 I

! Young produced Adult mortality \

Q F L

0 6L--1I-I L0I .Il 0 1 I t- _ 4 01__"

t-- I-=-

f 2 Young produced" Adult mortality ,.- L II, _ _

"I L- -

3 Young produced 0 1

_ Adult mortality . L L- - "- L U"L L..

4 Young produced \3 - __ L4 ___

Adult mortality - = L-. =. = =

5 Young produced ai a t to _I0 k" 0" Adult mortality L.- L_

= L..I L..- L._

6 IYoung produced n n 0 l0 (OIi I' Total young produced .- , * " - * ". "-* - ".1 Final Adult Mortality - . -

Note: Adult mortality (L = live, D = dead), SB = split brood (single brood split between two days), CO = carry over (otfspring carried over with adult during transfer).

Concentration:

% Mortality: 0.

Mean Offspring/Female: Z1,

% Reduction from Control: !,.9 I I Pa e 95 of 102 Indapendetrview by Kelley E. Keenan Initials: 1C-SOP ATI4 - Exhibit AT14.1, revision 06-01-11

Page 4 of 6 0ETS Z ErMMW 1ý~Wcmmnn1lSu ..

Species: Ceriodaphniadubia CdNaCICR #: 211-1 1200 mg NaCI/L Survival and ReproductionData Replicate number Day 1 2 3 4 5 6 7 8 9 10 2

Adult mortality Young produced f j. \

C0

' . L.- ' . ____ .. _ -

Adult mortalityI 1-- I -- "I I" "

3 1 Young producedI Adult mortality L. j ...

1. -

CI)I

.'_ \- '\

e. I I I'- '---

I 4 Young produced ____ ____ ____ 1___ _

Adult mortality ___1 __- It*_____

5 Young produced 1 LAT I LA 1 1I 6 [Young producedC)cC 0 )

Adult mortality \_ I ___ \-- V._

°-

7 Young produced t t -

Total young produced * . ' '

_

_ _

Final Adult Mortality \Z 9

\--.I I_-- _ I Note: Adult mortality (L = live, D = dead), SB = split brood (single brood split between two days), CO = carry over (offspring carried over with adult during transfer).

Concentration: I

% Mortality: 67.

Mean Offspring/Female: 8. 1

% Reduction from Control: "12.11.

1400 mg NaCIIL Survival and Reproduction Data Replicate number Day _ 11 2' 3 4 5 6 7 8 9 10 1 Young produced C01)O 0O, 0 A 0 0 o JI1.,,i0L..

l)

Adult mortality k-1..- \--- %-- \_ _- _. _

2 Youngproduced IL i 0IrF110 ' -I 3_J Young produced __

__ Adult mrtality __1 __ 0__ _ _ __ _ _

4 Young produced Adult mortality .. f ,.1~-I [ L J_- 0k- -JL-- 1I 0 I E%...-_

6 5 __.

Young produced Yugproduced __

C* I -

7 Young produced 0 C ~ ~ ~ .Q.0,-

Total young produced o . o ' i 0 6 Final Adult Mortality t X.- X?W * -_ T. _..

Note: Adult mortality (L = live, D = dead), SB = split brood (single brood split between two days), CO = carry over (offspring carried over with adult during transfer).

Concentration:

% Mortality::7.

Mean Offspring/Female: I0.

I% Reduction from Control: 1'11. 21.7.

Se 96 of 10 SOP AT14 - Exhibit AT14.1, revision 06-01-11

Ini*L~en Initials: r:eview 2yKelley E.Keenan V____

i* CDC t )

O:ETS (51 Environmental Testing Solutions, Inc.

Control Verification of Ceriodaphnia Reproduction Totals 1000 mg NaCMIL i5.

m Re plicate number Total Re plicate number Total Day 2 3 4 5 6 7 8 9 10 Day 2 3 1 4 5 6 7 8 9 10 m 1 0 0 0 0 0 0 0 0 0 1 0. 0 1 0 0 0 0 0 0 0 0 0 10 0 0

C, 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 4 5 5 4 4 3 4 4 4 4 4 41 4 3 4 3 3 4 4 4 4 4 4 37 5 9 9 13 10 10 11 12 9 10 10 103 5 9 9 11 10 10 9 11 10 10 10 99 6 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0 7 15 13 16 14 14 14 16 13 16 15 146 7 12 14 13 13 15 12 14 14 14 13 134 Total 29 27 33 28 27 29 32 26 30 29 290 Total 24 27 27 26 29 25 29 28 28 27 270 600 mg NaCI/L 1200 mg NaCI/L Re licate number Re licate number Tota Day 1 2 3 4 1 5 1 L. 7 8 9 10 Day 1 2 3 4 5 6 7 8 . 10 Total I 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 -0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 4 4 5 4 4 5 4 4 4 4 3 41 4 3 2 0 0 0 3 0 2 0 0 10 5 10 9 12 10 10 11 12 12 10 9 105 5 0 0 4 1 4 0 3 1 2 2 17 6 0 0 0 0 0 0 0 0 0 0 0 6 2 0 0 0 0 5 0 0 0 0 7 7 15 15 14 15 16 14 16 13 15 13 146 7 6 5 5 4 7 0 6 5 3 6 47 Total 29 29 30 29 31 29 32 29 29 25 292 Total 11 7 9 5 11 8 9 885 8 81 800 mg NaCI/L 1400 mg NaCI/L 0mg Da0 a Replicate number Total Re plicate number Total Day 1 2 3 4 5 6 7 8 9 10 Day ! 2 3 14 5 6 7 8 9 10 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 4 0 0 0 0 1 0 0 0 0 0 0 4 4 3 4 4 5 5 4 4 4 4 41 5 0 0 0 2 0 0 0 0 0 0 2 5 11 10 11 12 11 10 10 10 10 9 104 6 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0 7 15 14 14 14 13 IF 15 13 16 15 144 7 0 2 0 0 3 0 0 5 Total 30 27 29 30 29 30 29 27 30j28 289 Total 0 2 0 2 1 0 0 3 0 0 8 File: CdNaCICR_110811.xlsx Entered by: J. Surpner Reviewed by:

ur M 0ET 7amr 4 .

n io m na etn oui nI c

Ceriodaphniadubia Chronic Reference Toxicant Test EPA-821-R-02-013, Method 1002.0 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Test number: CdNaCICR #127 Test dates: November 08-15, 2011 Concentration Replicate number Survival Average reproduction Coefficient of Percent reduction from (mg/L NaCI) 1 2 3 4 5 6 7 8 9 10 (%) (offspringlfemale) variation (%) control (%)

Control 29 27 33 28 27 29 32 26 30 29 100 29.0 7.6 Not applicable 600 29 29 30 29 31 29 32 29 29 25 100 29.2 6.2 -0.7 800 30 27 29 30 29 30 29 27 30 28 100 28.9 4.1 0.3 1000 24 27 27 26 29 25 29 28 28 27 100 27.0 6.0 6.9 1200 11 7 9 5 11 8 9 8 5 8 100 8.1 25.7 72.1 1400 0 2 0 2 1 0 0 3 0 0 100 0.8 141.9 97.2 Dunnett's MSD value: 1.766 MSD = Minimum Significant Difference PMSD: 6.1 PMSD = Percent Minimum Significant Difference PMSD is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Lower PMSD bound determined by USEPA (10d' percentile) = 13%.

Upper PMSD bound determined by USEPA (9 0* percentile) = 47%.

Lower and upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET Interlaboratory Variability Study (USEPA, 2001a; USEPA, 2001b).

USEPA. 2001a, 2001b. Final Report: Interiaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes I and 2-Appendix. EPA-821-B-01-004 and EPA-821-B-01-005.

US Environmental Protection Agency, Cincinnati, OH.

File: CdNaCICR_110811.xisx Table populated from associated 'Verification of Ceriodaphnia Reproduction Totals" spreadsheet.

Spreadsheet entered by: J. Sumner Reviewed by:

Statistical Analyses SETS

!1 EnvronmentNi stsngSoluthflim .

Cenodaphnla Survival and Reproduction Test-Reproduction Start Date: 1118/2011 Test ID: CdNaCICR Sample ID: REF-Ref Toxicant End Date: 11/15/2011 Lab ID: ETS-Envir. Testing Sol. Sample Type: NACL-Sodium chloride Sample Date: Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Ceriodaphnia dubia Comments:

Conc-mg/L 1 2 3 4 5 6 7 8 9 10 D-Control 29.000 27.000 33.000 28.000 27.000 29.000 32.000 26.000 30.000 29.000 600 29.000 29.000 30.000 29.000 31.000 29.000 32.000 29.000 29.000 25.000 800 30.000 27.000 29.000 30.000 29.000 30.000 29.000 27.000 30.000 28.000 1000 24.000 27.000 27.000 26.000 29.000 25.000 29.000 28.000 28.000 27.000 1200 11.000 7.000 9.000 5.000 11.000 8.000 9.000 8.000 5.000 8.000 1400 0.000 2.000 0.000 2.000 1.000 0.000 0.000 3.000 0.000 0.000 I lUil*llgllll. UIIL{dlI*IUIIIIUU 1-Tallea Isotonic Conc-mg/L Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Mean N-Mean D-Control 29.000 1.0000 29.000 26.000 33.000 7.624 10 29.100 1.0000 600 29.200 1.0069 29.200 25.000 32.000 6.211 10 -0.259 2.287 1.766 29.100 1.0000 800 28.900 0.9966 28.900 27.000 30.000 4.143 10 0.129 2.2l7 1.766 28.900 0.9931

1000 27.000 0.9310 27.000 24.000 29.000 6.048 10 2.590 2.287 1.766 27.000 0.9278

1200 8.100 0.2793 8.100 5.000 11.000 25.667 10 27.065 2.287 1.766 8.100 0.2784

.1400 0.800 0.0276 0.800 0.000 3.000 141.912 10 36.519 2.287 1.766 0.800 0.0275 Auxiliary Tests Statistic Critical Skew Kurt Kolmogorov D Test indicates normal distribution (p > 0.01) 0.92959 1.035 -0.0957 0.26595 Bartlett's Test indicates equal variances (p = 0.29) 6.19973 15.0863 Hypothesis Test (1-tall, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnetts Test 800 1000 894.427 1.76577 0.06089 1605.2 2.98148 9.8E-45 5, 54 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point mglL SD 95% CL Skew IC05 932.105 76.6855 729.9 1004.59 -2.1990 IC10 1008.57 17.9052 955.121 1019.19 -3.1403 IC15 1023.97 6.4276 1010.14 1033.74 -0.1062 1.0-IC20 1039.37 6.02078 1026.79 1048.8 -0.0754 0.9-lIC25 1054.76 5.68364 1042.67 1063.88 -0.04001 0

. 1100.95 5.21302 1090.81 1110.54 0.0871 0.8-IC40 IC50 1131.75 5.41741 .1121.38 1141.94 0.1709 0.7:

0.6 C00.5-0.41 I .3-0 S00 1000 1500

.Dose mg/L Dose-Response Plot 1-tail, 0.05 level of significance File: CdNaCICR_11081 I.xlsx Entered by: J Smiler Reviewed by:

Page 99 of 102 Indeilsend: eview by Kelley E. Keenan Initials: ______

Ceriodaphniadubia Chronic Reference Toxicant Test D WControl Limits Determination v1 *( Environmental Testing Solutions, Inc.

m State and USEPA Laboratory Laboratory USEPA USEPA Test number Test date 7-day IC, CT S Control Limits SA Warning Limits 5

L&M Control Umits S&", Warning Limits So Control Limits CV S(g/L NaCI) (g/L NaCI) Cr-ns ar+2s r . C+ s~ cr-sj -sL

-. n+ s,,.

c CT- S,. CT + S.. a-CT.SA CT + SAM.

1 07-13-10 1.07 2 08-03-10 1.06 1.06 0.01 1.05 1.08 0.09 0.98 1.15 0.18 0.88 1.24 0.48 0.59 1.54 0.66 0.40 1.72 0.01 3 09-14-10 1.07 1.07 0.01 1.05 1.08 0.09 0.98 1.15 0.18 0.88 1.25 0.48 0.59 1.55 0.66 0.42 0.41 1.73 0.01 4 10-05-10 1.08 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.73 0.01 0.41 5 10-05-10 1.08 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.74 0.01 6 11-02-10 1.09 1.07 0.01 1.05 1.10 0.09 0.99 1.16 0.18 0.89 1.26 0.48 0.59 1.56 0.67 0.41 1.74 0.01 7 12-07-10 1.06 1.07 0.01 .05 1.09 0.09 0.99 1.16 0.18 0.89 1.26 0.48 0.59 1.56 0.67 0.41 1.74 0.01 0.41 8 01-18-11 1.06 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.74 0.01 0.41 9 02-08-11 1.06 1.07 0.01 1.05 1.09 0.09 0.99 1.16 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.74 0.01 0.41 10 03-08-11 1.05 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.73 0.01 0.41 11 04-05-11 1.06 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.73 0.01 0.41 12 04-05-11 1.07 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.73 0.01 0.41 13 05-03-11 1.07 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.73 0.01 0.41 14 06-07-11 1.05 1.07 0.01 1.04 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 1.73 0.01 15 07-12-11 1.06 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 16 08-09-11 1.06 1.07. 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.89 1.25 0.48 0.59 1.55 0.66 0.41 1.73 0.01 17 09-13-11 *1.06 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.88 1.25 0.48 0.59 1.55 0.66 OA1 1.73 0.01 18 10-04-11 1.07 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.88 1.25 0.48 0.59 1.55 0.66 OA1 1.73 0.01 19 10-11-11 1.06 1.07 0.01 1.05 1.09 0.09 0.98 1.15 0.18 0.88 1.25 0.48 0.59 1.55 0.66 0.40 1.73 0.01 20 11-08-11 1.05 1.07 0.01 1.05 1.08 0.09 0.98 1.15 0.18 0.88 1.25 0.48 0.59 1.54 0.66 0.40 1.73 0.01 Note: 7-d IC5 = 7-day 25% inhibition concentration. An estimation of the concentration of sodium chloride that would cause a 25% reduction In Ceriodophnio reproduction for the test population.

CT= Central tendency (mean IC25 ).

S = Standard deviation of the IC,, values.

Laboratory Control and Warning Limits Laboratory control and warning limits were established using the standard deviation of the IC,, values corresponding to the 10th and 25th percentile CVs. These ranges are more stringent than the control and warning limits recommended by USEPA for the test method and endpoint.

S,., = Standard deviation corresponding to the 10 percentile CV. (SASS= 0.08)

SA.s = Standard deviation corresponding to the 25h percentile CV. (Si,5 = 0.17)

USEPA Control and Warning Limits S&T7= Standard deviation corresponding to the 75e percentile CV. (S*,, = 0.45)

S.= Standard deviation corresponding to the 90" percentile CV. (SAao= 0.62)

CV = Coefficient of variation of the IC4 values.

USEPA. 2000. Understanding and Accounting for Method Variability in Whole Effluent Toxicity Applications Under the National Pollutant Discharge Elimination Program. EPA-833-R-00-003. US Environmental Protection Agency, Cincinnati, OH.

FM CdNaCICR_ 11811.,1 EntaM by. J n Re',, by.

lETS Page 5 of 6 Species: Ceriodaphniadubia CdNaCICR #: M.1 Daily Chemistry:

_ _[_ _ _Day

_ __ __ _ _ _ _ _0 1 2 Concentration Parameter "

pH(.. t* 0S I-A 1 9900 (gmhos/cm)

Conductivity t ,, l , "

CONTROL Alkalinity (mg CaCO 3/L) w4.1 Hardness (Mg CaC0 3/L) c~

Temperature "N* fit

_ __ _ ( 0 c) TA_ __ __ _ _ _ _

pH(S.U.)

pn 3su)

".t

,.~ 1.,l L \. *,1 A",

r~**r* qq 600 mg NaCi/L Conductivity 1o 600 mgNa~l/L(jimhos/cm) 0L Temperature (0c)_14ATA_(oc) ". _ t -, -.

pH (s.U.) "1,01 1-.01 to 1.1,,.121.

4 "~q 800 mg NaCVL Conductivity P+60 G40 (grnhos/crn)

Temperature IS.0 *2A.& -MSA 1 .

1000 mg NaCI/L Conductivity (oc)hos/cm) _ *00 Temperature zo .."__.-_

___(0c)_I__S 0 .0( "

1200 mg NaCVL Conductivity L.O

. ~ L . .. ... . . . . . .. . ... .

Temperature 260~*

____I___ ( 0 c) z~

DH (S.U.)

DO (mg/L) 1400 mg NaCI/L Conductivity :bo II +-0f 1.6.qq0 [ -.

(jimhos/cm) I Temperature (0C)

STOCK IConductivity 1 10.5=

I (Amhos/cm)

Initial 1 Final I Initial 11 Final I Initial 11 Final 4 it

,'q .9 w.* SOP AT14 - Exhibit AT14.1, revision 06-01-11 P e 01 ofIn ewyelley E. Keenan Initials:W

Page 6 of 6

ETSrn Species: Ceriodaphniadubia CdNaCICR #: 121 Day 3.4 5 6 Concentration Parameter pH (S.U.) AbIS..0.'o Conductivity (gmhos/cm) '30t 3 to 309 CNRL Alkalinity A~mlm CONTROL (mg CaCO3 L)

Hardness (mg CaCO 3/L)

Temperature - ti" -M.0 ck %It

.,L -'.* %.c. -..

, L. .s\ x** %

pH (S.U.) 10AM80 3O 600 mg NaCI/L Conductivity iý"30 (5eo (gmhos/cm) .75(1 Temperature -4.0 2LSZ IS .. S L I

I I

800 mg NaCl/L 1000~~D mg aCIL pHo(S.U.)

Conductivity (1IChosncm)

Temperature

_o__

PH (S.U.)

dutiit L* 00 V

__IN__

6.61 T*.-

24

.09L.

Pro

.15o so 13-03 1,04_____

I I (jimhos/cm)

Temperature LOS 7--4

{

',1 oO a

I 1000 mg NaCIIL Conductivity I 1400 mg NaCI/L Conductivity -0

-(-0-c)-

I Temperature "2.qh Initial 11

-j Final Ilb Initial II Final 2q.1-S Ilb - Initial II 7

Final Ilb -

2.4.

Initial II

"-S A Final II lb ii lb - lb -

Parage enit 1.0J tewoe f12 E. Keenan SOP ATI4 - Exhibit AT14.1, revision 06-01-11 alsend wt y Ke&lley

PERMII-IEE NAMEIADDRESS (Include Facility Name/Location if Different) NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) FMJOR Form Approved.

Name TVA - SEQUOYAH NUCLEAR PLANT DISCHARGE MONITORING REPORT (DMRJ (SUBR 01) OMB INo. 2040-0004 Address P.O. BOX 2000

- -_ (INTEROFFICE OPS-5N-SSQ. -TN0026450 I103 G F - FINAL SODDY - DAISY, TN 37384 PERMIT NUMBER I DISCHARGE NUMBER LOW VOL WASTE TREATMENT POND Facility TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY MONITORING PERIOD EFFLUENT I YEAR! MO DAY I YEAR MI DAY ATTN: Brad Love From 1 11 1 To T T-30L 11ii NO DISCHARGE [ --

NOTE: Read1 inqtructinne hafnm rn'mnhafinn thin fnr, PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO. FREQUENCY SAMPLE EX OF TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS PH SAMPLE

7 8 12 0 13 /30 GRAB MEASUREMENT 1 00400 1 0 PERMIT ** 6 9 SU THREE/ GRAB EFFLUENT GROSS REQUIREMENT MINIMUM MAXIMUM WEEK SOLIDS, TOTAL SUSPENDED SAMPLE * *10 12 19 0 2/30 GRAB MEASUREMENT 00530 1 0 PERMIT 30 100 MGIL TWICE/ GRAB EFFLUENT GROSS REQUIREMENT MO AVG DAILY MX MONTH OIL AND GREASE SAMPLE <9 <12 19 0 2 / 30 GRAB MEASUREMENT 00556 1 0 PERMIT 15 20 MGIL TWICE/ GRAB EFFLUENT GROSS REQUIREMENT MO AG DAILY MX MONTH FLOW, IN CONDUIT OR THRU SAMPLE 1.149 1.279 03 ... 0 30/30 RCORDR TREATMENT PLANT MEASUREMENT 50050 1 0 PERMIT Req. Mon. Req. Mon MGD SEE RCORDR EFFLUENT GROSS REQUIREMENT MO AG DAILY MX PERMIT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT NAME/TITLE PRINCIPAL EXECUTIVE OFFICER i Certify under penalty of law that this document and all attachments were prepared under my " TELEPHONE DATE direction or supervision in accordance with a system designed to assure that qualified personne John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the person or persons who manage the system, or those persons directly responsible for gathering the Vice President information, the information submitted is, to the best of my knowledge and belief, true, accurate, 423 843-7001 11 12 09

,,euoya,, Site Vie President 1

and complete. I am aware that there are significant penalties for submitting false information, SIGNATURE OF PRINCIPAL EXECUTIVE including the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER YEAR MO DAY TYPED OR PRINTED I CODE LUMMEIN I UAEXPL NAI'I.I IUFUt- ANY VIULA I IUNS (Reference all attachments here)

An O&G sample was collected and sent for analysis on 11/16/2011. The sample results were 17 mg/L O&G. SQN shipped the backup sample collected on 11/1612011 for confirmatory analysis and the results were < 6.3 mglL. SQN believes the original sample was contaminated by the laboratory. The daily maximum reported is an average of the two samples collected on 11/16/2011.

Page 1 of I

~PA EPA FOrE (REV 3153) 3320-1 (REV Form J~ZU-1 3199) Previous editions Previous may be editions may be used used Page 1 of 1

PERMIIT-EE NAMEIADDRESS (Include FacilityName/Location if Different) NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR Form Approved.

Name TVA - SEQUOYAH NUCLEAR PLANT DISCHARGE MONITORING REPORT (DMR)

(SUBR 01) OMB No. 2040-0004 Address P.O. BOX 2000 j -NTEROFFICE E OPS-5N-SQN). NI TN026450 110 G I F- FINAL SODDY - DAISY TN.337384 PERMIT NUMBER NDISCHARGENUMBERI RECYCLED COOLING WATER Facii TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY MONITORING PERIOD EFFLUENT I YEARI MOIDAYI Froml -Il 1 11 01 I YEA IX1RM AY I NO DISCHARGE ATTN: Brad Love ToNOTE:Read instructions before completinq this form.

PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO. FREQUENCY SAMPLE EX OF TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS TEMPERATURE, WATER DEG. SAMPLE 04 CENTIGRADE MEASUREMENT 04 00010 1 0 PERMIT * *******

REPORT DEG C CONTIN CALCTD EFFLUENT GROSS VALUE REQUIREMENT DAILY MX UoUS TEMPERATURE, WATER DEG. SAMPLE

CENTIGRADE MEASUREMENT 04 00010 Z 0 PERMIT 30.5 DEG C CONTIN CALCTD INSTREAM MONITORING REQUIREMENT DAILY MX UOUS TEMP. DIFF. BETWEEN SAMP. & SAMPLE 04 UPSTRM DEG.C MEASUREMENT 00016 1 0 PERMIT 5 DEG C CONTIN CALCTD EFFLUENT GROSS VALUE REQUIREMENT DAILY MX UOUS FLOW, IN CONDUIT OR THRU SAMPLE 03 TREATMENT PLANT MEASUREMENT 03 50050 1 0 PERMIT Req. Mon. MGD CONTIN RCORDR EFFLUENT GROSS VALUE REQUIREMENT DAILY MX UOUS CHLORINE, TOTAL RESIDUAL SAMPLE **19 MEASUREMENT 50060 1 0 PERMIT 0.1 0.1 MG/L Five per CALCTD EFFLUENT GROSS VALUE REQUIREMENT MO AVG DAILY MX Week TEMPERATURE - C, RATE OF SAMPLE CHANGE MEASUREMENT 04 82234 1 0 PERMIT 2 DEG C ******** CONTIN CALCTD EFFLUENT GROSS VALUE REQUIREMENTUOUS SAMPLE MEASUREMENT PERMIT REQUIREMENT NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my TELEPHONE DATE direction or supervision in accordance with a system designed to assure that qualified personnel John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the person or persons who manage the system, or those persons directly responsible for gathering the S 0 h ite Vice President information, the information submitted is, to the best of my knowledge and belief, true, accurate, 423 843-7001 11 12 09 Sequoyah Site Vice President and complete. I am aware that there are significant penalties for submitting false information, SIGNATtfRE OF PRINCIPAL EXECUTIVE TYPED OR PRINTED including the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER YEAR MO DAY TYPED ORIPRINTED CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

No Discharge this Period EPA Form 3320-1 (REV 3199) Previous editions may be used Page 1 of 1

PERMIT-EE NAME/ADDRESS (Include FacilityName/Location if Different) NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR Form Approved.

Name TVA - SEQUOYAH NUCLEAR PLANT DISCHARGE MONITORING REPORT (DMRJ (SUBR 01) OMB No. 2040-0004 Address P.O. BOX 2000

- (INTEROFFICE OPS-5N-SQ.N)- TN026450 I 110 T F -FINAL SODDY - DAISY TN 37384 PERMIT NUMBER DISCHARGE NUMBER RECYCLED COOLING WATER Fa.cility TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY MONITORING PERIOD ]EFFLUENT I YEAR M DAY I YEARI MO I DAY From l 1111 101I NO DISCHARGE **

ATTN: Brad Love To IlI 11 30 NOTE: Read instructions before completinq this form.

PARAMETER QUANTITY OR LOADING [ QUALITY OR CONCENTRATION NO. FREQUENCY SAMPLE EX OF TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS IC25 STATRE 7DAY CHR SAMPLE 23 CERIODAPHNIA MEASUREMENT TRP3B 1 0 0 PERMIT 43.2

PERCENT SEMI COMPOS EFFLUENT GROSS VALUE REQUIREMENT MINIMUM ANNUAL IC25 STATRE 7DAY CHR SAMPLE

PIMEPHALES MEASUREMENT 23 TRP6C 1 0 0 PERMIT 43.2

PERCENT SEMI COMPOS EFFLUENT GROSS VALUE REQUIREMENT MINIMUM ANNUAL SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my direction or supervision in accordance with a system designed to assure that qualified personnel TpTELEPHONE DATE John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the person or persons who manage the system, or those persons directly responsible for gathering the eq aite Vice President information, the information submitted is, to the best of my knowledge and belief, true, accurate, 423 843-7001 11 12 09 Sequoyah Site Vice President and complete. I am aware that there are significant penalties for submitting false information, SIGNATUR-E OF PRINCIPAL EXECUTIVE including the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER YEAR MO DAY TYPED OR PRINTED CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

No Discharge this Period EPA Form 3320-1 (REV 3199) Previous editions may be used Page 1 of 1

PERMITTEE NAMEIADDRESS (Include Facility Name/Location if Different) NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR Form Approved.

Name TVA - SEQUOYAH NUCLEAR PLANT DISCHARGE MONITORING REPORT (DMAfRJ (SUBR 01) OMB No. 2040-0004 Address P.O. BOX 2000

.... ..INTEROFFICE OPS-5N-SQN) TN0026450 118 G F - FINAL SODDY - DAISY1N._373 84 . PERMIT NUMBER DISCHARGE NUMBER WASTEWATER & STORM WATER Facilty. TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY MONITORING PERIOD EFFLUENT I YEARI MO . DAY I YEARI MO DA ATTN: Brad Love Froml 11 11 101 1 Tol il 1 30 NO DISCHARGE l

NOTE: Read instructions before completinq this form.

PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO. FREOUENCY SAMPLE EX OF TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS OXYGEN, DISSOLVED (DO) SAMPLE **

MEASUREMENT 00300 1 0 PERMIT 2 MGIL TWICE/ GRAB EFFLUENT GROSS REQUIREMENT MINIMUM WEEK SOLIDS, TOTAL SUSPENDED SAMPLE 19 MEASUREMENT 19 00530 1 0 PERMIT 100 MG/L TWICE/ GRAB EFFLUENT GROSS REQUIREMENT DAILY MX WEEK SOLIDS, SETTLEABLE SAMPLE 25 MEASUREMENT 00545 1 0 PERMIT

MUL ONCE/ GRAB EFFLUENT GROSS REQUIREMENT DAILY MX MONTH FLOW, IN CONDUIT OR THRU SAMPLE 03* **

TREATMENT PLANT MEASUREMENT 03 50050 1 0 PERMIT Req. Mon. Req. Mon. MGD ONCE/ ESTIMA EFFLUENT GROSS REQUIREMENT MO AVG DAILY MX BATCH SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT SAMPLE MEASUREMENT PERMIT REQUIREMENT NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my ,d') 7 TELEPHONE DATE direction or supervision in accordance with a system designed to assure that qualified personnel John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the person or persons who manage the system, or those is, topersons the bestdirectly responsibleand for gathering information, the information submitted of my knowledge belief, true,the accurate, StViePresident 423 843-7001 11 12 09 Sequoyah Site Vice President and complete. I am aware that there are significant penalties for submitting false information, SIGNATUFOF PRINCIPAL EXECUTIVE I including the possibility of fine and imprisonment for knowing violations- OFFICER OR AUTHORIZED AGENT AREA NUMBER YEAR MO DAY TYPED OR PRINTED I CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

During this reporting period, there has been no flow from the Dredge Pond other than that resulting from rainfall.

EPA Form 3320-1 (REV 3199) Previous editions may be used EPage 1 of 1

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