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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 prepared under 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 gathering the 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, re c

sident SoIoyah Nuclear Plant Enclosures cc (Enclosures):

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

PERMITIEE NAME/ADDRESS (Include Faciliht Name/Location if Different)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000

..L.N1TEROFFICE OPS-5N-SQN SODDY - DAISY TIN 37384 Facility TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON CQUNTY ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

DISCHARGE MONITORING REPORT (DMR)

TN0026450

ý 101 G PERMIT NUMBER IDISCHARGE NU ER MAJOR Form Approved.

(SUBR 01)

OMB No. 2040-0004 F - FINAL DIFFUSER DISCHARGE EFFLUENT NO DISCHARGE [j j NOTE: Read instructions before completinq this form.

MONITORING PERIOD YEAR MO I DAY I YEAR I MO DAY From 11 11 011 To 1

11 3o 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 PERMIT Req. Mon.

MGD CONTI RCORDR EFFLENT ROSSREQUIREMENT 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 I

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

. (

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)

Previous editions 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 12/14/2010 12/22/2010 12/29/2010 01/04/2011 01/11/2011 01/18/2011 01/25/2011 02/02/2011 02/08/2011 02/15/2011 02/22/2011 03/01/2011 03/08/2011 03/16/2011 03/23/2011 03/30/2011 04/06/2011 04/08/2011 04/20/2011 May 2011 June 2011 July 2011 August 2011 Sept 2011 October 2011 11/20/2011 11/29/2011 6

0 0

20 0

0 22 0

0 18 45 21 100 0

0 0

0 100 0

0 0

100 100 100 23 10 10.5 26 13 22 9.5 23 10 9

23 10 11 11 11 11 12 15 15.5 16 12/07/2010 12/14/2010 12/22/2010 12/29/2010 01/04/2010 01/11/2010 01/18/2010 01/25/2011 02/02/2011 02/08/2011 02/15/2011 02/22/2011 03/01/2011 03/08/2011 03/16/2011 03/23/2011 03/30/2011 04/06/2011 04/08/2011 04/20/2011 11/20/2011 11/29/2011 0

0 0

0 23 10 10.5 26 13 22 9.5 23 10 9

23 10 11 11 11 11 12 15 15.5 16 1-25-545 1-25-545 1 -ISV-24-1234 1-25-545 1-25-545 1-25-545 1-ISV-24-1234 1-25-545 1-25-545 1-25-545 1-25-545 1-25-545 1-ISV-24-1236 1-ISV-24-1236 1-ISV-24-1234 1-ISV-24-1234 1-ISV-24-1236 1-ISV-24-1234 1-1 SV-24-1236 1-1 SV-24-1236 PB RS WE WDT PB RS CR WDT PB MJW MLW PB PB WE MLW MLW MLW HMW WAW/PB PB No Samples Collected No Samples Collected No Samples Collected No Samples Collected No Samples Collected No Samples Collected 0

0 0

0 14 14 18 0

14 14 1-ISV-24-1236 1-25-545 BC

PERMITTEE NAME/ADDRESS (Include Facility Name/Location if Different)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 AINTfEROFFICE OPS-5N-SQdN)

SODDY - DAISYTN. 37384 Facilty TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

DISCHARGE MONITORING REPORT (DMR)

E TN0026450

][101 T

PERMIT NUMBER

]

DISCHARGE NUMBER]

YEA M

MONITORING PERIOD YEAR MO DAY I IYEARI MO DAY Froml 111 11 1 01] Tol 111 11 3in MAJOR Form Approved.

(SUBR 01)

OMB No. 2040-0004 F - FINAL BIOMONITORING FOR OUTFALL 101 EFFLUENT 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 0

1 / 180 COMPOS CERIODAPHNIA MEASUREMENT 23 TRP3B 1

0 PERMIT 43.2 PERCENT SEMI COMPOS EFFLUENT GROSS REQUIREMENT MINIMUM ANNUAL IC25 STATRE 7DAY CHR SAMPLE

>100.0 23 0

1/180 COMPOS PIMEPHALES MEASUREMENT 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 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 0*ic 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, SIGNFIýE OF PRINCIPAL EXECUTIVE ncluding 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)

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

EPA Form 3320-1 (REV 3199)

Previous editions 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 (Pimephales promelas)

Daphnids (Ceriodaphnia dubia)

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%

Ceriodaphnia dubia: IC_5 > 100%

UV treated Outfall 101: Pimephales promelas: 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 Intake 110711 0610 11-07-11 1339 0.6

<0.10 11-09-11 1140 11-07-11 0610 11-09-11 1140 11-08-Il 0655 to 11-10-11 1148 101 11-09-11 1230 0.9, 0.7'

<0.10 11-09-11 0555 11-11-11 1139 11-08-11 0705 to 11-10-11 1148 Intake 11-09-11 0605 11-09-11 1230 1.0

<0.10 11-11-11 1139 11-09-11 0605 11-11-11 1139 11-10-11 O655to 11-12-11 1149 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

Test Organisms:

Pimephales promelas Aquatox, Inc.

20.33 - 20.58 hours6.712963e-4 days 0.0161 hours 9.589947e-5 weeks 2.2069e-5 months old Ceriodaphnia dubia In-house Cultures

< 24-hours old

1. Source:

2. Age:

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 Moderately Hard Synthetic

3. Control / Dilution Water: Moderately Hard Synthetic

4. Number of Replicates:

4

5. Organisms per Replicate: 10

6. Test Initiation: (Date/Time)

Outfall 101 UV Treated Outfall 101

7. Test Termination: (Date/Time)

Outfall 101 UV Treated Outfall 101

8. Test Temperature: Outfall 101:

10 1

11-08-11 1235 ET 11-08-11 1220ET 11-15-11 1146ET 11-15-11 1130ET Mean = 24.8°C (24.3 - 25.1 °C) 11-08-11 1011ET 11-15-11 1006ET Mean = 24.9'C (24.8 - 25.2 0C)

Test Temperature: UV-Treated Outfall 101:

Mean = 24.80C (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 Piniephales promelas 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 Ceriodaphnia dubia 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/IC25 Page 6 of 102

TOXICITY TEST RESULTS (see Appendix C for Bench Sheets)

2.

Results of a Ceriodaphnia dubia 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 = I 00/LC50: TUc = 100/IC 25 Page 7 of 102

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

3.

Results of a Pimephales promelas 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)

(%replicate number)

(%Eflen) 1

]

2 3

]

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)

Pimephales promelas November 08 - 15, 2011 1245 7-days KC1 0.78 g/L Ceriodaphnia dubia November 08 - 15, 2011 1000 7-days NaCI 1.05 g/L Page 8of 102

0 PHYSICAL/CHEM ICAL

SUMMARY

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

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 CaCO3)

Chlorine (nig/L)

Control 24.8 24.6 7.7 7.5 8.02 7.80 311 62 89 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 10.8%

24.9 24.6 7.7 7.4 7.99 7.77 297 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.

24.9 24.7 7.7 7.4 7.99 7.79 259 43.2%/

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'-

210 100.0%

25.0 24.7 7.8 7.4 7.94 7.85 188 73 77

< 0.10 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 Intake 24.9 24.6 7.9 7.5 7.98 7.87 187 73 77

< 0.10 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 Control 24.8 25.0 7.7 7.7 8.02 7.97 311 62 89 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 41 24.9 25.0 7.7 7.8 7.99 7.99 259 z

43.2%

,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 25.0 24.9 7.7 7.8 7.97 8.03 205 86.4%

24.9 -

25.0 24.8 -

25.1 7.6 -

7.8 7.6 -

8.0 7.84 -

8.09 7.94 -

8.09 201 210 100.0%

25.0 24.9 7.8 7.8 7.94 8.04 188 73 77

< 0.10 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 Intake 25.0 24.9 7.9 7.8 7.98 8.08 187 73 77

< 0. 10 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)

Pimnephales promnelas Ceriodaphnia dubia Average 24.8 24.9 Minimum 24.3 24.8 Maximum 25.1 25.2

(D 0

NJ PHYSICALJCHEMICAL

SUMMARY

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

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 CaCO3) (mg/L CaCO3)

Chlorine (mg/L)

Control 24.9 24.6 7.8 7.5 7.99 7.76 310 61 88 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 10.8%

24.9 24.7 7.8 7.4 7.99 7.78 296 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 25.0 24.6 7.8 7.5 7.97 7.86 206 86.4%

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 -

211 100.0%

25.1 24.6 7.9 7.5 7.94 7.84 190 73 77

<0.10 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 Intake 25.0 24.6 7.9 7.5 7.98 7.86 187 73 77

< 0.10 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 Overall temperature (C)

Pimephales proinelas Ave rage 24.8 Minimum 24.4 Maximum 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. Pitnephales promelas 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. Ceriodaphnia dubia 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. Pimephales promelas None

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

1. Pimephales promelas None

2. Ceriodaphnia dubia 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 IC25 values in g/L KC1 or NaCl.

2.

Standard Toxicant: Potassium Chloride (KC1 crystalline) for Pimephales promelas.

Sodium Chloride (NaCI crystalline) for Ceriodaphnia dubia.

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 1 st 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 09/09/1998 0.003 0.031 0.011 09/10/1998 0.014 0.060 0.021 09/11/1998 0.013 0.055 0.019 09/12/1998

< 0.001 0.044 0.015 09/13/1998

< 0.001 0.044 0.015 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 08/21/1999 0.022 0.068 0.024 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 0.020-12/17/2000

< 0.0036 0.025 0.020 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 01/05/2002 0.0042 01/06/2002 0.0042 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 01/12/2003

<0.0035 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 08/03/2003

<0.0050 08/04/2003

<0.0050 0.058 0.020 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: 11 IV£ Fax:

828-350-9368 Fil dniiainNumber Flow

,,::.*.,:,.,~*g*.

.*o,,,..,z...

prab/Comp.

Collection DateTime Volume (MGD)R F d DentictionVolle (Mark as Appropriate)

Yate lieif

Yes, No Trace 1Sg.

wyen 4

SQN-TNT-TOX Comp f, 4/7//7/0 o

"(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

Sample temperature performed using Sample Receiving Thermometer: SN 6338.

I:ETS 4'.Wue Whole Effluent Toxicity Sample Receipt Log-to Date Time Received Received Sample Project Sample Sample name and description State Comments received received by from temperature (*C) noumber

_ 1, 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 S*-*

5 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 /// 6 9-.

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.

.m 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 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 S11-09-11 1007 K. Keenan Fed - Ex 0.5 7507 111109.19 WashiPgtoneWrP NC ShearonHarrisPlantNC 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 Ptject Name: Sequoyah NP Toxicity P4. Number: N/A Facility Sampled: Sequoyah NP NPDES Number: TN0026450 Environmental Testing Solution, Inc.

351 Depot Street.

Asheville, NC 28801 Phone:

828-350-9364 Delivered By (Circle One):

FedEx UPS Bus Client E

e Other (specify):

General Comments:

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

_z~~r Sample Custody - Fill In From Top Down

'4 Sep&S i zr SO-f'.es tlec-ew'i es Relinquished By (Signature):

Date/Time Received By (Signature):

Date/Time

-5A~tA1 I

g o

C_

X/

i i O

E

~~~it t I__

~t 1

~

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 mm E I~

r~

Tho~m~

Page _ S

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

Whole Effluent Toxicity Sample Receipt Log ian in As Date Tone Received Received Sample Project Sample Sample name and description State Comments received received by from temperature (*C)*

number 1-I1-I I 0956 K. Keenan Fed - Ex I.1 7496 111111.01 Bladenboro WWTP NC I1-11-11 0956 K. Keenan Fed - Ex 0.6 7497 111111.03 Progress Energy Carolinas - Cape Fear S.E.U taie NC I11III 0956 K. Keenan Fed - Ex 0.5 7498 111111.04 W

V*est negyw - allnas - ae erP S ptemItk NC I 1-11-11 0956 I Keenan Fed-Ex 0.5 7498 111111.04 CiPr ofgGastuniae

-Darlas ww-rP FearS._F._NC I1-1-I1 0956 K. Keenan Fed-Ex 0.7 7499 111111.05 UnitedWater-EnfieldWWTP NC 1-1111 0956 K. Keenan Fed-Ex 0.5 7500 1 111111.06 Progress Ener*y Carolinas - Lee Steam Plant NC 11 0956 IC Keenan Fed-E.

0.6 7501 111111.07 Duke Enrgy Corporation - Marsh.all Steam Station NC I -I1I11 0956 K. Keenan Fed -Ex 0.7 7502 111111.08 United Water - Scotland Neck WWTP NC I1-1111 0956 K_ Keenan Fed-Ex 0.4 7511 111111.09 ProgessEnery Carolinas-ShearonHarrisF-&E-Center NC S1-11-11 1003 K. Keenan UPS 0.3 7490 111111.10 Microbac-Rockingh WWTP NC 11-11-11 1003 K. Keenan UPS 0.3 7491 111111.11 Microbac - Roseboro WWTP NC I1-t1-11 1003 K. Keenan UPS 0.3 7492 111111.12 Microbac - J.P. Stevens - Wagram WWTP NC Il-Il I1 1032 K. Keenan Dash Courier 1.0 7508 111111.13 OWASA -Mason Farms WWTP NC 1I-11 11 1235 J. Sumner M.rphy's Courier 27 7512 111111.14 Duke'oer Corporation - McGuire Nuclear Station - 1.9 Compusite NC I1-11-11 1220 J. Sumner TVA Courier 2.111.7 7477 H111111.15

,TVAT-uoahNuerPlant-Oudll 101 I1-11-11 1220 J. Sumner ITVA Courier, 2.0 7477 111111.16 TVA - Sequoyah Nuclear Plant -Intake TN SOP G4 - Exhibit 04.2, revision 06-29-09

pr 0ETS SEngionmentalTestingSolutl Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephales promelas A.

Page 1 of 6 Client: Tennessee Valley Authority Facility: Sequoyah Nuclear Plant 006 NPDES #: W44 0 *60 TW o021'so *0 Project#:

I f"t County: ai4*w4

.3 Outfall: 101 Dilution prep aration 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 Drying information for weight determination:

Transfer bowl information:

pH =

1.-0 S.U.

Date / Time in oven:

V-\\

C)

Temperature = 2q.

G°C Initial oven emperature:

O0 'C.

Average transfer volume:

Date / Time out of oven:

.t,.

il2.A Final oven temperature:

t o..

Total drying time:

"-L4geS Daily feeding and renewal information:

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

%.k, hOG*

,h tl0l~l ttea..

-0+-IIA 2

'i-f'-l

.~.4..J +/-...6

_k I'.o I-I1 WO'l1.

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 It..

IG,,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: Pimephales promelas Client: TVA / Sequovah Nuclear Plant, Outfall 101, Non-treated Date:

Ii-os "I Survival and Growth Data I

I 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 1 I /C '

o /0 3

10 10 10 (0 10 10 10

/

1

'0 10 10 /0 Io D o 10 0 1 o 1 10 M I0 C) 10 to 5

10 10 0o /0

/0 /o 10 1o

/0 t0 /0 /0 6_I0

/0 to /0

/O'

"/C

'0 /0

/0

/0 t0 o

to I~1 to lo If/0 to 1

0 A = Pan weight (mg)

Tray color code::

LI;jt\\ 1p,.

Analyst:

MM

'I.'i.

%SA..

d l

14*

I-461.L 4'

.t 11

.(-4 130; 14.%

, 14.s(.

,q, Date:

t1O Tr -

I

.I I

Il B = Pan + Larvae weight (mg)

Analyst:

M_

Date:

1.1q.1 l o.1%

Ilq4. ui

"..V3

.1 t..1%

.7 2.4.4,, 7.o.'AL 2..

Iq.o 5.4o Z,. rl vA.i4 C - Larvae weight (mg) = B - A Hand calculated.

bA q sl

(

'r 1.1 I

-1.S 1,

.13

i.,q4 16-1,

&M, 1.60 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 Average weight per Percent reduction Initial number of from control()

01 0 larvae (mig)

I I

~

I a

i 0.1,41 1,:-7, 0-1LIO Al.7.

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.

i I

I Comments:

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

Page 3 of 6 IETS t

lutions, Inc.

Species: Pimephales promelas 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

/0 I

0 o

0

/0 10 t0 -0 30,0 t0o 0

10

/0

,0 1O

/0

'0

/0

,0 10

/0

/0 /0 2

J0

/Q

/0

'o 10

'00

/0

/0to 3

to to

/C6 1'0 1C)

IQ C)

/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__

t0 o0 lb 10

(

,I

, to l

lo A = Pan weight (mg)

Tray color code::

L.,-,v Analyst: _*.4*

M'.

(,t is.44 I

1. to 4

-.4 14.1.,

I t.4 lt.'

l1.52 It1.". I -.4 14.l1 Date:

\\__"__

B = Pan + Larvae weight (mg)

Analyst: ________.

4______

Date:

11.____

it_._-1_

U li 1,o.o*$

UA1.t, Z1L.,.

7L.4

-A4

'L.Ll

-4.5 e Z4.-

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

Al0

.1 14 4,l

" 0 ll.

QO A

Analyst:

Weight per Initial number of larvae (mg)

C / Initial number of larvae Hand calculated.

IIP qO 4 I

QJ°o* % J Analyst:

-o 6"

g)

[*

()

O" ci 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.

iIm ens Comments:

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

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

it-w "I\\

Day 100% Intake Y

Z AA BB 0

1L C

0 /10 1

/0 10 2

/0 I0 t

_________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 e34 8l

,ng penin revie~w~y Kelley E. Keenan Initials:

SETS ni Environmental Testing Solutions, Inc.

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 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses 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 1-a berof 1

m-u l (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:

Dunnet's MSD value:

PMSD:

Intake:

Dunnetl's MSD value:

PMSD:

0.1249 16.2 0.0914 11.9 MSD =

Minimum Significant Difference PMSD =

Percent Minimum Significant Difference 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.

Lower PMSD bound determined by USEPA (10th percentile) - 12/0.

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, PlnAe.tg14 i*94 Kelley E. Keenan Initials: _

File: sqn10lIlOlldata.xlsx Entered by: J. S ueer Reviewed by:

T

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 0.6' C: 0.5 0 0.4 0.3 0.2 0.1 0

C i1-tail, 0.05 level of significance o

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

SET'S F

Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated November 08-15, 2011 Pimephales promelas Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Daily Chemical Analyses Project number:

7477 Concentration Parameter Da 0 Da I Da 2 Da 3 Da 4 Da 5 Da 6 Initial IFinal IInitial IFinal Initial IFinal iInitial IFinal IInitial I

Final IInitial IFinal IInitial IFinal Control Alkalinity (mg/L CaCO3)

Hardess (mg/L CaCO3)

Temperature (C 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 PH (SU 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 DO m 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 10.8%

Conductivi hos/cm 303 300 300 296 293 295 293 Tem erature C) 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 PH (SU 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 DO m 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 21.6%

Conductivi hos/cm 286 286 283 284 283 2

281 Tmeaue24.9 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 Tern eratnre C

2.9 2.6 2.1 2

2.0 2.8 2.1 2.7 2.9 2.5 2.9 24 2.8 2.5 43.2%

Conductivi hos/cm)

Tern erature C) 25.1 24.8 25.1 24.8 25.0 24.7 25.1 24.5 24.9

24.

24.9 24.6 24.8 24.7 8.%

Conductivit (imhos/em) 0 0

0

ý 0

1 0

Temperature OC)2.

246

2.

245 2.1 2.1

2.

247

2.

245 2.1 46 2.845 Conductivit (phos/cm) s1018

ý191 16 100%

Alkalinity (mgfL CaCO3)

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

Tern erature C)

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 24.6 100% Intake Alkalinity (mg/L CaCO3)

Hardness (mg/]L CaCO3)

Total Residual Chlorine (mg/L)

-Temperature *CC)2.1 2.1 2.1

2.

50 47 49 45 2.1 2.1 2.1 2.1 2.1 2.

File: sqn101_110811chem.xls Entered by: C. Johnson Reviewed by:

4

I I

I p

Page 5 of 6 "ETS I EnvirnmentalTesting Solutions, Inc.

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

Date:

ll-Le* \\

I Dav I

Analyst Concentration IParameter Conductivity himhos/ern)

CONTROL Non-treated 4

Alkalinity (mgn CaCOSU) a Hardness (mc CaCOd/L)

Temperature (0C0 a

ý pH (S.U.)

DO (m-/L) 10.8%

Conductivity (Aimhos/cm)

Temperature (0C0 4

I PH (S.U.)

t

~

DO (mg/L) 21.6%

Conductivity (amhos/cm)

Temperature (°C) 43.2%

Conductivity imhos/cm)

Tem Merature (0C) 300o 300 I

00oo IS-,

1qo

+.o L*-

.la L.Q L.

1S.0 0 00 4-S t¢.

2S. k U-0 ne Tl nH (S.U)'

.4.M DO (rag/L) 86.4%

Conductivity Temperature (°CQ 111.1.9 7 -pH (S.U.)

q Conductivity Alkalinity 100~~_(mg CaCO3 "L)

Hardness T-(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 100%

Intake Conductivity (gmilos/cm)

Alkalinity (Mg CaC0 31L)

Hardness (Mg CaCo 3/L)

TR chlorine (mg/L) 40.10 Temnerature (ofl I

2(4.

I.-

I 1i 0

Initial II 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 oETS i

EnvironmentalTesting Solutions, Inc.

Species: Pimephales promelas Client: TVA / Seaunuvah Nuclear Plant. Outfall 101. Non-treated Date:

it I I

Day I

Analyst F 3

I 11 A r~ir I A Pe-r" Concen-Parameter tration I

pH (S.U.)

DO r(rag/L)

Conductivity (umhos/cm)

CONTROL Non-treated Alkalinity (me CaCOJL) 34-0 I. bznvý II -6, I

q---"-I "A Hardness (me CaCO,/L) 4.1 0.

S

-LI.6 Temperature (0C)

II

,,A, a

a nH (S.U.)

II 3-4C. I

~

II pH (S.U...)

r m

DO (mg/L) 4A'#II 10.8%

Conductivity rI (gmhos/cm)

W-LA6 I sao 1 i-+¶~o Temperature (0C)

M.0 II a

a ~

pH (S.U.)

DO (mg/L) 21.6%

Conductivity

[

Qirnhos/cm) a. [(,

I U

0. 04 elTemnerature (°C*

L¶.%I I

Temperature (Q a

DH (S.U.)

4.iO 9 10 I

^%.LA II

_ A to 00 ~i DO (mi/L) 43.2%

Conductivity (umhos/cm)

IS I-I0

1 Temperature (MC) a 4

nH (S.U.)

.t II % '.

OH (S.U.)

DO (m&/L) 86.4%

Conductivity

(,jmhos/em)

=A0(o "LAp O.f Ua3TO

" a.j 1q.

,9

_i A

J S1 IQ Wo"

.3

'20 Temperature (0C0 "t., \\ II I

I ~

pH (S.U.)

w0 rII A~-

DO (mg/L)

I "-9~

100%

Conductivity (tgmhos/cm)

Alkalinity (mg CaCO 3IL)

Hardness (mg CaCO 3/L)

TR Chlorine (mg/L)

Temperature ('C) p DH (S.U.)

I n ii I

DO (mgiLn 2,1-195 ij

t 7Q-S

-. z-...-

100%

Intake Conductivity mgrhosfcmn Alkalinity nm CaCO3/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: Ceriodaphnia dubia Client: Tennessee Vialley Authority Facility: Sequoyah Nuclear Plant NPDES #: TN0026450 Project #:

NL131 County: Hamilton Outfall: 101 Dilution preparation information:

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 organism source 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 between:

Inlocation:

n Culture board:

I A- 011 A.i oato:2 Replicate number:

1 2 I3 4 1 5 7 1 9 1-9 1 10 YTbth Culture board cup number. I I'. I O1 "

". Z,32.

YWT batch:

I

1-1153I 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 H__

I-04 *-

if A

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.

.\\ I0 I. -

,,*L 4

11-12.11 1O__

a1-_

_._n I_ ___

__L 111111.16" ii -

I&

iIt l,tS t

l.16 h

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 3rd Broods:

1 007..

10:7*

> 80%

NOEC I007.

" Mortality:

0,()I,

- ! 20%

LOEC 5 1067.

Mean Offspring/Female:

?. I 21.. (a 15.0 offspring/female ChV j

1*ii7.

%CV:

5-0.

".37.

< 40.0%

IC25

>_too'.

Pl#AVed#k )fe1 by Kelley E. Keena n Initials:

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

lETS Page 2 of 7 Species: Cerlodaphnia dubia Client: TVA / Seqluoyah Nuclear Plant. Outfall 101 CONTROL-1 Surv Date:

Wi-1A ival and Reproduction 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

Young produced 0

i 1

I T3-Iac Adult mortality L_

\\.

)

C

%)I*.-

4 Young produced

'S

'[%

Adultmortality 11],_i0__

5 oung produced 1 16iQ1.S2%0 I

I C) 1 C) C%

C:)1 6

Young produced D

0 0

Adult mortality I*1**I**]**

I*";***I" I'

Total young produced W

It V-%

5.g t

o0

?

s o*

Final Adult Mortality

_C7

\\.

s_

ordBos.

X for 3' Broods 5c-.

x.

t-5Z7.-"*

-7 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 and Reproduction 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

C-'

i'-1' L_-

I V__I 4

Young produced Adult mortality j1 l

+/- __

Young produced 1(0l 17 t Il (31 Adult mortality

[_*

1, L..l.

U C7*.

dut_

_ _ _L I

Young produce 0

0 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),

-cO an over (offspring carned over 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 Ir-nQtqa i_

Kelley E. Keenan

.1Initials:

JETS Page 3 of 7 Species: Ceriodadhnia dubla Client: TVA / Sequoyah Nuclear Plant, Outfall 101 CONC: 21.6%

Survi Date: _

_i_

ival and Reproduction Data 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 t

_V.

2 Young produced I

I]1

)

... [ Adult mortality t.

2 Youngproduced L

i.

.]

Q, Adult mortality 4

Young produced t

I C

L-l..

1

_ Adult mortality 5 Young produced il Zf 1YIIVp I

2 Z

Adult mortality 75-Young produced I" ti.

"0 [ 't.LiA I Ii I ~I aI l

1 I *T I

61 Young produced Id C 01 ]0IOD[

0 Q

_ Adult mortalty j

J

~

j'..

-I' cI'I~

Is, TYoung produced t-

.16 I'll Ill Is i

Total young produced 3

5

o 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*

CONC: 43.2%

Survival and Reproduction Data Replicate number Day 1

2 3

1 4 1 5 6

7 8

9 10 1

Young produced 0_

1 0' I

5 (J

j(

0 Adult mortality L

I.--

L L_

2 Young produced 0 -'

)

C)

Adulmortality 3

Y Youngproduced C,

G 0

1 In Adult mortality 4

Young produced 4

(11 1 WL S__

Adult mortality L

[

L.

U.

L-J 5

Young produced It 2JJJ1 I

I~I.I..

Adult mortality M

6

Yung produced 01_

L i =1 K Adult mortality

%--~

J \\-j\\-'-

=- t--

7 Young produced I I 20 IS i

I I*

" I *t Total young produced 34 Z

1 3

3

.0

.3S 5-6 3S

'Final Adult Mortality

%t L.

]a L.

LL.

I I

I Note: AdUlt mortality (L = eive, D = dea), StB = split broad (single brood split between two days), Cu = ca Pa

_e 43 of 10 e

In 'penu*,K reviewoy Kelley E. Keenan Iýnitials:

ovseprJtspnnrg canm over with adult durng transfer).

Concentration:

% Mortality:

G ".

Mean Offspring/Female:

Ab.I

% Reduction from Control-I:

-A.17a SOP AT 11 - Exhibit ATI 1.2, revision 06-01-11

SET

ndmnnmlmdTe.*fla SrUnt,.

Page 4 of 7 Species: Ceriodaphnia dubia Client: TVA / Sequoyah Nuclear Plant, Outfall 101 CONC: 86.4%

Suryi Date:

It-04F11 ival and Reproduction Data rs~v D

nInt t nulmber Cp, *[L IlU*Ubl 1

2 3

4 N

6 7

S 9

10 Da 14 2

3-Young produced Q

10 0

1 0 L*)

0 Adult mortality

\\

1 I '..

2 Young produced 1C1)f 0

0 0

SAdultmortality

\\.-

l

____ltI_._EKI L...

3 Young produced I p

p O T

O 0

I_

W I 0I

_ Adult mortality I 4_i..I LI

-I I

I-I 4

Y oung produced It16.l I l t

- lt T l I l i [ L

_ Adultmort*itylI I-1

1+-I L--I 5

Young produed I c

.l I l

. I A '2. 1 10 2. lO I Adult mortality

'-I _'I I

"-V---

6 Adult mortality

__~

- K ]

5I I.

7 Young produced 14l 16 La za

-VI Total young produced 810 J,q s.A 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 and Reproduction 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 I A

L i 6 0

Adult mortality r

L_

l*._

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

Qi L-I 10 "2 C 3___

T o ta l y o u n g p r o d u c e d j

4 3 0 %.36 3

1' I

15 S-S 3 -6*s 3 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: Ceriodaphnla dubia Client: TVA / Sequoyah Nuclear Plant, Outfall 101 CONTROL-2 Survi Date:

Itok"It.

ival and Reproduction Data Replicate number Day 1

2 3

4 5

6 7

8 9

10 Adult mortality I.L..

L

~_

2 Young produced C

2 Young produced 0

0 c

0 0 0

1 0

_J Adult mortality L****

I__ I'-_

I_

I__

L I

I Adult mortlity I L-L I

I[Ll %.- Il~

I Young produced

=o 31

_ Adultmortality 5

[ Young produced I

b I Adult mortality I

-i

-J ---_

kII 6

Young produced 1%

[

0.

t 94 I

1,5

%-6 6 Yunpoue Adult mortality *'-,,

.. J

'~_'-"

I '-

'7

[Young produced t

,t j t*

t I

t¢ I I

Total young produced 71 1 11 7

n

- 3 I 2*

2.7 Final Adult Mortality

Ž1. ']+/-1 X for 3r Broods Note: Adult mortality (L - live, D - dead), SB split brood (single brood split between two days), CO = carry over (offspring canTied over with adult during transfer).

Concentration:

% Mortalit:w Mean Offsprinlemnale:

21.I

" I CONC: 100% Intake Survival and Reproduction Data Replicate number Day 1

2 3

4 5

6 7

8 9

10 1

Young produced C) 0n Adult mortality 1-L

)

L JLJ L(

L'(

L(

.2 Young produced 13 C

0 1~ 0 1 0 I0 I0 I

Adult mortality I

L_

L L",

I k

I 3

Young produced l

0.

0 1*CO ls Adult mortality ILl LI L'-

Young produced 1 %

1 1

IJ.L 7F q

Adult mortality I_

I I

Q I

T I

5 Young produced Adult mortality L

I --

I L IL I 7

]Young produced l'*

Z o

\\%C lo'I ttA I

Total young produced T

I j

I 3

I 2' Final Adult Mortality L..

?

I

'- I k

k-.--

A.-

,. -A-U t o

Iy k

ý

.10 O...-

M

.I.--

-,a oc 111 I

Concentration:

% Mortality:

01-Mean Offspring/Female:

33.. L

% Reduction from Control-2:

o.

70.

SOP ATI 1 - Exhibit AT 11.2, revision 06-01-11 erfla

'! La l'94Kelley E. Keenan Initials:

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated November 08-15,2011 CD~

CD

-X(S 00ETS

)

E ontralTesthVSol-t loerV IS Control-I Verification of Ceriodaphnia Reproduction Totals Day 2

'licate number Total 1 8 1 9 1 10 0

0 10 101 0

0 0

0 1t 0

flIAI l

lf0 tt 0O0OI 0

4 5

3 3

3 4

5 3

3 36 5

9 11 11 10 1I 10 10 9

9 10 100 6

0 0

0 7

16 13 15 15 14 15 12 16 17 13 146 Toa 0

2L7 2 29 28 26 30 29 26 282 10.8%

Day Re plicate number Total 21 2

3 4

5 6

7 8

9 10 1

0-0 0

0 0

2 0

0 0

3 0

0 0

4 4

5 4

3 4

5 4

4 4

4 41 5

ll.

10 11 10 12 11 11 13 10 10 109 6

0 0

0 l, 7 17 N17 15 1

14 19 116 15 19 17 167 Total 32 321 30 32 23 31 3

2 33 31 317 21.6%

Dy IRe*

ticate number 8

0 Total Day 1

2 3

4 5

6 7

8 9

10 1

0 0

0 2

0 0

0 3

0 0

0 4

5 4

4 4

4 5

4 4

J 4

q41 5

12 10 12 11 10 10 12 10 13 10 110 6

0 0

0 7

18 20 15 20 18 16 15 19 195 15 164 Total 34 31 302 32 31 35 33 31 29 315 43.2%

DyRep~icate number 9

0 Total D a 1

2 1 3 4

5 1 6 7

8 9

1 I

0 0

0 2

0 0

0 3

0 0

0 4

4 6

4 4

4 5

5 4

5 5

46 5

12 12 10 10 10 11 10 12 12 12 111 6

0 0

-0 0

0 0

0 7

18 20 15 20 18" 1 7 1 '5 19 19 18 179 Total 34 r8-2-9 34 r2 r3 SU 3.1

_21 35 36 86.4%

Replicate number Total Day 1

2 3

4 0

6 7

8 9

1 0 2

0 0

0 2

0 0

0 3

0 0 -0 0

0

-0 0

0 0

4 5

6 5

4 4

4 5

4 6

5 48 5

12 13 11 11 12 0

1 11 13 13 118 6

0 0

0 7

19 16 18 19 18 20 20 17 19 18 184 Total 36 35 34 34 34 34 37 32 3L 36 350 100%

DyReplicate number Total D

1Y 1

2 3

4 5

6 7

8 9

10 1

0 0

0 2

0 0

0 3

0 0

0 0- 0 0

0 0

0 4

5 4

5 5

6 5

5 4

4 4

47 13 12 13 12 10 12 12 11 11 12 118 6

0 0

0 7

16 19 21 19 18 19 18 20 18 18 186 Total 34 35 39 36 34 36 35 35 33 34 351 Control-2 DyRep)licate number Total DY 1 213 4

5 6

7 8

9 10 H

0 0

0 1

0 0

0 3

0 0

0 4

3 3

4 4

4 3

3 3

33 5

10 10 9

10 12 10 10 10 10 11 102 6

0 0

0 7

14 14 14 13 14 16 15 13 14 14 141 Total 27 27 27 27 30 29 28 26 27 28 276 100% Intake DyReplicate number Total Day 1

2 3

4 5

6 7

8 9

10 1

0 0

0 2

0 0

0 3

0 0

0 "0

0 4

4 3

4 5

5 5

3 5

4 4

42 5

11 13 13 12 12 11 12 11 11 11 117 6

0 0

0 7 o 17 20 15 1

' 15 17 17 15 19 19 173 1 Total 132 136 132 361 32 133 32 31 34 34 332 File: sqn101_110811data.xlsx Entered by: J. Surrper Reviewed by:

E S

(D Environmental Testing Solutions, Inc.

on GJ Concentration TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated November 08-15, 2011 Ceriodaphnia dubia Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1002.0 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Project number:

7477 Replicate number 4

S 6

Survival Average reproduction

(%)

(offspring/female)

Coefficient of variation (%)

Percent reduction front control (%)

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:

Dunnett's MSD value:

PMSD:

Intake:

Dunnett's MSD value:

PMSD:

1.946 6.9 1.156 4.2 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) = 13%.

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 0rF Tn 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 REntered by: J. Suker Initials:

.,ee Rýeedb

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

Avbo nmeM&Th 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.

30 25

&20 15 10 5.

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

1,.1111111ýý

.I 1-tail, 0.05 level of significance u

8 8

GO C (C4 Nt 0

m 0

0 C~

C Plr@Ppn 1+95 119; Kelley E. Keenan Initials: _

File: sqnlOl_110Blldata.xlsx Entered by: J. S u er Reviewed by:

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

Project number:

F5 I

Concentration IParameter Day0 I

Day I

Day 2 Initial I Final I Initial I

Final I

Initial I

a PH (IS 8.021 8.05 7.871 7.90 DO (mp/L)

Conductivity (pmhos/cm)

Control L

Alkalinity (mg/L CaCO3) 80 77 87 Hardness (mg/L CaCOM) 4 DOm L

10.8%

Conductlvi hos/cm rem erature C

DO m

21.6%

Conductivi hos/cm DOm L

43.2%

Conductivi hos/cm 86.4%

Conductivi hos/cm Condnctivi hos/cm 100%

Alkalinity (mg/L CaCO3)

Hardness (mg/L CaCO3)

Total Residual Tem rature

°C DO m

Conductivi hos/cm 100%

Intake Alkalinity (mg/L CaCO3)

Hardness (mg/L CaCO3)

Total Residual Chlorine (m*L)

Tem erature C

.... ~I

~

emperature IL) 24 20 4,82,,24.

24.9 2,5 Z

4 4.51 24.9 z5.:z 2 24 249 24.81 25.11 24.91 25.01 PH 7.87 802

.05 7.871 7.87 80 8.071 7.971 7.96 803.06 80 8.03 79 DO (mg/L) 7.9 79.779 7

7.

6 7.7 7.6 76.8775 7.67.

10.8%

Conductivit (pho/em) 303 300 300 296 293 295 293

_________Temperature CC) 4 24.9 252.20 24.8 29 24.8 24.82.

249 50 249 24.

8 25.0 2.

pH (SU 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 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 (junhos/em) 286 28,6 28(3 284 281 278 281

_______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) 7.88 8.02 8.06 7.89 7.88 8.0 8.10 800 7.97 85 7.96 804.05 79 DO (n91L) 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 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 DO (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%

Conductivity fjsmhos/cm) 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 7.7179 7.

8.01 7.7 7.8 7.71 7.

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

Alkalinity (nsg/L CaCO 3 )

73 37 Hardness (mgfL CaCO 3 )

77 77 Total Residual Chlorine (mg/L)

'01

<0.10

<0.1

_______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 8 8.0 80 7.991 8.121 7.871 8.101 8.041 8.22 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 7.71 7.7 Conductivity

(~Imhos/cm) 18 18 18 17186ý 1886 100% Intake Alkalinity (mg/L CaCO 3 )

73 37 Hardness (mg/L CaCO 3 )

79 77 Total Residual Chlorine (mr/L)

I

<0.1 I01 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:

ETS

1 Tnlll~l S~I~q4aI~nc Page 6 of 7 Species: Ceriodaphnia dubia Client: TVA / Sequovah Nuclear Plant, Outfall 101 Daily Chemistry:

Date:

1l1-0i9 I

Day I

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

DO (mg/L)

Conductivity (pmhos/cm)

CONTROL Alkalinity (g

CaC03/L)

Hardness (mg CaCOv/L)

Temperature (°C) 21.6%

Conductivity ze*O PH (S.U.)

DO (mg/L) 43.2%

Conductivity (larnhos/cm)

Temperature ('C) 1 pH (S.U.) wQ 86.4%

DO (mgIL)

Conductivity (jiihos/cm)_

Temperature (°C)

DH (S.U.)

1-90 DO (m/L) 1.11 100%

Conductivity IQ--

&+/-mhos/cm)______

Alkalinity (mg CaCO3/L)

Hardness (Mg CaCo3/L)

TR chlorine (mgfL) 0.i Temnerature (°C)

-2&.8 i

a pH (S.U.)

I-.A1 100%

Intake DO (mg/L)

IN0 Conductivity I £f1-Alkalinity (mg CaCQ3/L)

Hardness (mg CaCh 3/L)

TR chlorine (mg/L) 40..0 Temperature (°C)

U-.0 U

U

-initial 11 Final I

Initial 11 Final I

Initial 11 Final i

IntiAns:

v i~ q& Kelley E. Keenan Initials: _

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

iETS Species: Cerlodaphnia dubia Client: TVA / SeQuoyah Nuclear Plant, Outfall 101 Date:

11-&,111 I

I m

Day I

K I

Day I

I I

"5 I

5 I

Analyst Concen-jParameter I tratin pH (S.U.)

DO (mg/L)

.v Conductivity (gmhos/cm)

-I-CONTROL

.[Alkalinity (m, CaCOUL&

Wv i

_ _4 4

Hardness (mg T aCt3/L)

Temnerature ('C)

I nn(5~ TI~

PH (SU).

DO (ma*/U)

DO (mg/L) 10.8%

Conductivity (iimhos/cm) a.

Temperature (*C)

I I* ~-Ii-nlli (* T T (S )

.U.

U DO (mg/L)

-r 21.6%

Conductivity (umhos/cm)

Temerature (°C)

IpH (S.U.)

0.10, I DO (mg/b) 43.2%

Conductivity (gTmhos/cm)

F1,3_

_________Temperature

('C) 1-1-

01.

S.0 njH (*q.TTa pH (S ).

DO (mr_/LI DO (mWL) 86.4%

Conductivity (gtmhos/cm) a.

Temnerature (°C)

-emperaure

C nit (Q T T "a PH (SU).

DO (mp/L)

I 100%

Conductivity (jmhos/cm)

Alkalinity Hardness TR Chlorine (mg/L)

Temoerature (0 )

I I

I pH (S.U.)

DO (rag/L) 100%

Intake Conductivity (jimhos/cm)

Alkalinity (Mg CaCO3/L)

Hardness (mg CaCO3/L)

TR chlorine (mg/L)

Temperature (QC) m I

I-I I

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

ETS

, EnvlronmentalTesting Solutions. Inc.

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

Species: Pimephales promelas JW Page 1 of 6

%I Client: Tennessee Valley Authority Facility: Sequoyah Nuclear Plant NPDES #: WN0,0269A 114 0

,7J.0 Project #:

-'"AV County:

4eA A-*.vb*J Outfall: 101 Dilution pre aration information:

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:

no LAN Randomizing template:

&.0l Date and times organisms t -0"1-11 06.0 Incubator number and were born between:

shelf location:

Organism source:

A,1W U-rck ac i-k".i Artemia CHM number:

Drying information for weight determination:

Transfer bowl information:

pH =

1. 1 C0 S.U.

Date / Time in oven:

Temperature =

I 0C Initial oven temperature:

t' Average transfer volume:

Date / Time out of oven:

0. I ' l, -

Final oven temperature:

Total drying time:

Daily feeding and renewal information:

IVOU 0

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 0

,I l*.

W -#I A..

1 1 Vz. 7o A,. lnI 1 0. 01 j luai**,

IV04 11A I

I I-Og-110

.0 0 At

iqoo A.s A1 l

nl '. o

t 1*i,Oa.

-L llo q-6+

11 1

,I -,o'i ohoo j-

'4 to0 4AL.

t\\LS '

,,,t0,.o t

,\\,

l.-o.. il -*o -il 5__

O L

L

..4N i.t1

_, Is 1111 11.16, 11-o0 0 6

1 V 14-d Al.

WL

\\I.

M' I L200 :ý I t, I4 ii11111 I It-01-lI C o n tro l in fo rm atio n :

\\........

A cceptance criteria S u m m ary of test en dp o in ts:

% Mortality:

n p7.

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 r yew'Kelley E. Keenan Initials: _ _ _

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

"ETS EnvironmentalTesting Solutions, Inc.

Page 2 of 6 Species: Pimephales promelas 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 10

/0

/0 (0

10

/0-10

/0

(,) /0

/o

/0 3

60 0

C

/0 I*

o j

10 10 c

10 10 1" )

10 4ID 10 to

/0 /

1 0

/M

/I 1(3 t0. /6 (0

5 0

1 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 123.S'o..

1'/4 1,.t Date:

1.t*lk.-L C = Larvae weight (mg) = B - A H and calculated.

6.. 1 "h i(

I'.

&" I a. 21 - i. q '4 1.2.(e 6 S*

1. 2.i 2 1.S/

,..1 -

Analyst:

-/-

Weight per initial number of larvae (mrg)

= C /Initial number of larvae Hand calculated.

Analyst:

rio 0*

0' 0*

~0 0

~0 tY 4ý*L"t3%

01 01 0*

AA" No Average weight per-Percent reduction o.

initial number of from control ()

I 0 2i larvae (mug)

I a

h P a I

(.617-1..'7.

0.101

. 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

I 0ETS EnvZonmentaITtng Solutions, nc.

Page 3 of 6 Species: Pimephales promelas Client: TVA / Sequoyah Nuclear Plant, Outfall 101. UV-treated Date:

t1"O4-)l Survival and Growth Data Day 43.2%

86.4%

100%

10

10) 10

/

o c

Jto /0 o

to ID /

/Io /D 10

/0 JO 10

'0

,0 10 10 10 2

/0 /0 i

C

/0 C I00 0

10

/D

/0 10 f 0 23/

/O D O

10 '

0 1 r IO to IO C O /I

(

10

(

4 to

/( 0 /D

/

/0 Ib I0 VIA lo

/0

/0 10 5

it' 1(0 1 0

.tO C

D I O

) I )

(

/ (

/0 1 D 6

/0

/

/0

/0.

/ol0 o

/

00

/0 C0 7

I f3 C) 1(j t

to A0.-

1011 /D

/G I o A - Pan weight (mg) o' Tray color code::

l aA zt.

Analyst:

1__

__1_._2_

i 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)

Analyst:

M4 It-l*

A L..

t. 6.A Z.

Date:

1.1"'11

&ejs t -5uI 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 Hand calculated.

'O Analyst:

{*

O" r"

q*

'" )"

("o

)'

-A-

%19

\\ r-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:

PiRp AnRnv9?y Kelley E. Keenan Initials:

U SOP AT20 - Exhibit AT20.3, revision 04-01-09

I I

"ETS

.0 EnvlronmentafTesting Solutions. Inc Page 4 of 6 Species: Pimephales promelas Client: TVA / Sequoyah Nuclear Plant, Outfall 101, UV-treated Date:

I-04I-11 Day 100% Intake

__Y Z

IAA BB 0

tO00o o

10//

0 1

/D0 2

/0 3/(

4/

/0 5

to 1

o 6

7

/0 A f Pan weight (mg)

Tray color code::

Analyst:

kJ

s l

.1.50 Date:

tO4*-*t2 B = Pan + Larvae weight (mg)

Analyst:

2_Ot

_o

?4

(.q Date:

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

1.-1q,

Analyst: _

Al',

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_______

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:

l n ntTlvin Kelley E. Keenan Initials: ___=__

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

I EL toEnvironmental Testing Solutions, Inc.

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 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Prloect numbaer 7477

'a SNot for Compliance Aýssessrmnente.at Laboratlr 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 MeweightI Cdfideatetvalfatloa Weight/ blitni amber Me rslvat Me-welghtlInial Coerlent or Perceni.dsajl froai 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 Control B

10 10 13.89 21.07 7.18 0.718 0.724 4.7 0.718 100.0 0.724 4.7 Not applicable Control C

0 10 12.95 20.28 7.33 0.733 0.733 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 G

10 10 13.23 19.70 6.47 0.647 4100.0 0.672 6.6 7.

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 21.6%

O 10 10 14.69 21.98 7.29 0.729 0.703 6.7 0329 100.0 0.703 6.7 2.8 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 R

10 10 14.08 20.56 6.48 0.648 0682 11.3 97.5 0.665 133 8-1 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 0.835 100%

V 10 10 13.32 20.08 6.76 0.676 0714 11.3 676100.0 0.714 113 13 W

10 10 13.97 20.62 6.65 0.665 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 0.744 100% Intake Z

10 10 12.84 21.05 8.21 081 0.761 6.4 100.0 0.761 6.4

-5.2 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; Dunnetf's MSD value:

PMSD:

Intake:

Duonett's MSD value:

PMSD-0.1004 13.9 0.0575 7.9 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 determuned 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 Intehraboratuory Variability Study (USEPA, 2001a; USEPA, 2001 b).

USEPA. 2001a, 2001b. Finial Report: 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 "ETS I Env nmenti Testing r

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 0.6810 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 0.6 0.5 09'0.4

0.3 0.2 0.1 0_

1-tail, 0.05 level of significance CO4 1

o -O B

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

PM VW~~~

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 IC20

>100 0.9 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

0.5i

" 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'

7 Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 - UV-treated November 08-15, 2011 Pimephales promelas Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Daily Chemical Analyses 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 lHardness (mng/L CaCO3) 7 Total Residual Chlorine - WL

<0.1

<0.10

_______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:

III SI I

00ETS Environmental Testing Solutions, Inc.

Page 5 of 6 Species: Pimephales promelas Client: TVA / Seauoyah Nuclear Plant, Outfall 101. UV-treated Daily Chemistry:

Date:

11i04-1 Day 0

1 2

ýZ 2

0-0 1 IODT I CW 11 ICOT IeD CW 11_m Analyst Concentration IParameter PH (S.U.)

4.9r4-

+W)

-I-OMM-11 q-.1-3 I *A-A 11 431H DO (mr/Li CONTROL UV-treated Conductivity (jimhos/cm) 51 Alkalinity Hardness (mg CaCO3/L)

Temperature (°C)

Ts. 0 pH CSU..IA 10.8%

Conductivity (jgmhos/cm)

Temp erature (C)Q t~'

21.6%

Conductivity (4.mhoscm)

___________Temperature (0C)

).S.0-43.2%

Conductivity (pgnhos/cm)

_________Temperature (QC IIS.0 021o 1.0 4 4

,-:th I

-. ot (0 di

~ILU 11100 1 R

I m.o 7A L-I 'M 2S0

!".o pH (S.U.)

DO (mg/L) 86.4%

Conductvity Temperature (°C)

'L.

I.

Conductivity (g+/-mhos/cm)

Alkalinity 10M%

C 3/0 Hardness (Mg CaCO3IL)

TR chlorine (mg/L)

Temperature (0C Z.

Conductivity (jimhos/cm) 100%

Alkalinity i"

Intake (mg CaCO3/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:

0 "DnETS I1 EvrnmetlTsig olutions, tne..

Page 6 of 6 Species: Pimephales promelas 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 i

I I

I CONTROL UV-treated Hardness (me CaCOSL'i (Mg CaC03/1-)

Alkalinity

(*m CaCOe/YU Temperature (°C)

I j

10.8%

Conductivity Temhos/cm)

C emperature C0C)TU 1H (S.U.)

9 -~--I:

7' DO (mrg]L) 21.6%

Conductivity (umhos/cm)

Temperature (°C)

U S ~

pH (S.U.)

DO (mg/L)

I n

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) 9-100%

Conductivity

(;tmhos/c.)

Alkalinity (mg CaCO3/1.)

Hardness (mig CaCO3IL)

TR Chlorine (mg/L)

Temperature (0C)

I r

I I

II ii

.- 7 pH (S.U.)

DO (mg)L)

I

%.k II -

I,'9,1 II T'.W I 9F.F II q'*>

I

~ ~~~~~

if 1

I

+-

kT-

=

100%

Intake Conductivity (gmhos/cm)

Alkalinity (mg CaCO3/L)

Hardness (mig CaC03/L)

TR chlorine (nmg/L)

Temperature (°C)

-'5 ieI 2~.l N. 6 1 7.4L21 Fnl I Initil Thminal Initial Fina-l Initial 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 I

Analyst F7 Date analyzed Calibration:

Ic odide reagent:1 T--

IIIII, 1 Acid reagent:

TOI=L4, Iio

!I 0.10 mg=L, 1.00 mgaL S

s lope MRe~ference standard number

/

.N$-O 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 ND0 Cfl rn Loe o7 2

I 111~.06 PCs, Dmsdhiae.

Jxor'. 204a 001%

nnoio1 1VA S %n 101 LAX_______

111l1(.CTZ-3 640 1 q~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 control standard:

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 I

Reviewed byI V

Date reviewed SOP C8 - Exhibit C8.1, revision 06-01-11 Page 63 of 102

Page t

Page i

of _

S En TS 41 Environmental Testing Soluflonrn Inc.

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 Date analyzed

10. It Calibration:

Iodide reagent: I ' 7 I.4 Acid reagent: I*,I

/o I

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 control standard:

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 Duplicate sample precision:

Sample SampleSEI)e Sample characterstic Residual chlorine

%RPD = {(S - D) /[(S+D)/2]) x 100 number (mg (acceptable range = + 10%)

/ Duplicate D Z 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 rio,~x a,...t4,[,-

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.-

0.50 O.S KF G

t6.

o401.

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

iNP XIi Page _1 Page Iof

?.

ETS 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 Date analyzed Calibration:

Iodide reagent: LI*.*

IIZI Acid reagent:

I-S-4,4o 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.

I Laboratory control standard:

[

Reference standard True value (TV)

Measured value (MV)

% RS = MV / TV x 100 number (mg/L)

(mg/L)

(acceptable range = 90 to 110%)

-4,

0.50 o.0,, c

.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(

4-A 0.2000%

A lilt 14, icp-uit a..r peg.~

411cY4 l~llLIL ~0.

6.-.

4,

_____1 0.2 cor).-L14 AM 0351 Note: All samples were analyzed in excess of EPA recommended holding time (15 minutes) unless otherwise noted.

Laboratory control 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 Analyst f" 7,1-7 Date analyzed Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO3/L Titrate samples to pH = 4.5 S.U.

Time initiated j[qq Time completed I S I

J_."....

Duplcate sam pe precision:

Sample Alkalinity

%RPD Sample Sample ID volume Begin End Total IMultiplier (Ing CaCO3/L)

[(S -D) /[(S+D)/21) x 100 number (ml) ml ml I

ml.

M

/

(acceptable range 110%)

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 Sample alkalinity (B)

Measured spike value (MV)

% R= MV / SV x 100 I

I (ing CaCO3/L)

MV = A - B (acceptable range (mg CaCO3/L)

-75 to 125%)

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,.

3.*

10

/

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 Reviewed by:

E=

Date reviewed:

-"q" II Page 66 of 102 SOP C6 - Exhibit C6.1. revision 09-01-09

SEnvironmental Testing Soludons, on Page Page

2. of J; Time initiated [Id..IL,,

]

Time completed [

ZZ-Analyst 7Pkil'i,,-

I Date analyzed I i-, iq.I I

I Titrant nornmality and multinlier determinatioi Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO 3/L Titrate samples to pH = 4.5 S.U.

PH of Normality Normality (N) of H2SO4 pH Factor or Multiplier Deionized Titrant check Begin End Total

= (5 ml Na2 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 control standard:

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 Duplicate sample precision:

Sample Alkalinity

%RPD =

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

((S - D) /[(S+D)/211 x 100 number (ml) ml ml ml (acceptable range=L 10%)

I-d5*

jUgt.

6.0

'44

'f-1

1) 10-1

S M 4, Duplicate (B)

D j~,_____

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

_426

,7..

"7S 111112-01 o.,6 7.5

,5s 7C

,I I I 1 1-0 I ll11Z. 07---

Mitl~t VIC.7 IV* 1 1'.0 II Reviewed by:

Date reviewed: I I\\- 1411 Page 67 of 102 SOP C6 - Exhibit C6.1. revision 09-01-09

.j.

Environmental Testing Solutions, Ise.

Page

'If Page 3

of5 Analyst 3u Date analyzed 11..11 Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO3/L Titrate samples to pH = 4.5 S.U.

Time initiated Time completed

]

Titrant normaliy and multiplier determination:

pH of Normality Normality (A) of H2S0 4 pH Factor or Multiplier Deionized Titrant check Begin End Total

= (5 nil Na2CO3 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

Laboratory control standard:

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, U.

OI9 i.6%

Duplicate sa= e precision:

Sample Alkalinity

%RPD =

Sample Sample ID volume Begin End Total Multiplier (mng CaCO3/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)

(mg CaCO3/L)

(ml) ml il mi l

%'tS Isal 10:0 so 0~

t,0 i

$.J I'-5 16.1 l t 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%)

ZZO q

Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID (mi) ml mi ml Multiplier (Ing CaCO3/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 "U3 L.X S (

,i) 2 C0 odeAO.

3.

1 L

S.5 Itzo 1 lllI0M,-

.*0

r.

c.Ij

(.0_)_

_1_

111 a1-,0S Cit i..q a(,

=o000 I'i A11.&olI ujteq eALor

-L S

I&

'~aI.S C&1) i I

i i

Reviewed by:

I2ZZ Date reviewed:

I 711 -ii' Page 68 of 102 SOP C6 - Exhibit C6. 1. revision 09-01-09

3 nTeSo

)

EnvironmentalTestIng Solutions, Inc.

Page IZ Page _ __of 5

Time initiated [

[

Time completed LZI

]

Analyst 37 4 7b Date analyzed nI.i I

]

TifrnnI nn~rnmnlsu aNd inItln far,Ln'..,.~itt..

Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO 3/L Titrate samples to pH = 4.5 S.U.

pH of Normality Normality (N) of H-2S0 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)

Laboratory control standard:.

Reference standard True value Sample Alkalinity (MV)

%RS=MV/TVx 100 number (TV) volume

Begin, End Total Multiplier (rag CaCO3/L)

(acceptable range (mg CaCO3/L)

(ml) ml ml ml

=90 to 110%)

X Xts s ol' 100 100

g.

S 3

.o.q q7 qT. 0 Duplicate sam, 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 = k 10%)

Duplicate (B) lam..

C, S D

f-C Matrix spike recovery:

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

(mg CaCO3 L)-

ml mi 1 "m ml ml Xt-16s aj Soo 10 43 11,3 1

,06 Sample alkalinity (B)

Measured spike value (MV)

% R = MV / SV x 100 (rag CaCO3/L)

MV = A - B (acceptable range (mg CaCO3IL)

= 75 to 125%)

Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID m ) t.,%

ml ml ml Multiplier (Ing CaCO3/L)i 1.-1.4L 1lL oi T11-S7

-o\\

i,,1

-w io.&C i'.i 5.C CIO)

"6.)1w 1 )

s1'~.

of

~

It.C N i.

C CZI7 unnot

-ix

+/-

5W__ t7o% t.f

-3.3 I n o 0i.l ZI Iwo6 e.C 73 I11o.I U.

71 Z IC 3.4 73 I I nI I al. 67

-CIA,)

r8 loZ4 3.4 73

I 11aoI.22-C X'_

9"l.

3.C "73

3*

o,¢ 3*,l

&¢ 73 iiI I.o t

tok 5.

I I

I Reviewed by:

Date reviewed: I 11.14-W/

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

S

-* EnvironmentaliTesting Solutions, inc.

Page

'13 Page

.9 of 5 Analyst Date analyzed ý l

Tihrant nnriralift and,nultlnlt'r de'iprmlnatln Alkalinity (SM 2320 B)

Matrix: Water, RL = 1.0 mg CaCO3/L Titrate samples to pH = 4.5 S.U.

Time initiated Time completed

-...... no

.. and Tr

inadol, pH of Normality Normality (A) of [12SO4 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

/

OT.O__

b.

II.l~

Reviewed by:

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

Envronrne.nt Testtng Solutions In.

Page 73 Page I

of

.I Time initiated ois' Time completed rGo' Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO3/L Analyst

i 1

Date analyzed

11. 1.11 Titrant normalitv 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 -.0.0220)

= N x 1000 Laboratory control standard:

Reference standard True value Sample Hardness (MV)

% RS = MV / TV x 100 number (TV) volume Begin End Total Multiplier (mg CaCO3/L)

(acceptable range (lig CaCO3/L)

(ml) ml ml ml

=90 to 110%)

40 50 16.1 I,1 2 I2.

0o

00.8%

Duplicate sa le 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 I

Li I ~

Duplicate (B)

Z.

D0________

Matrix spike recover, :

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

-f(ing CaCO 3/L)

(ml) ml mi ml 0W 1

"I Io s

qI i*Io Ill) 111

4.

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 CaCO3/L)

TV = ND Blank (should be = 0 Mg CaCO3/L) 0.I 0

,0 D.o ii'S O.-0'\\-1i A Z,

-I__1 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 P tteef*4 &I1 02 8

ant is used, sample must be diluted.

Reviewed by:

Dtriw Date reviewed 0 -00 Ib SOP C7 - Exhibit C7. 1. revision 09-01-09

3 Environmentll Testing Solutions. In.

Page ?q Page 2

of_

Time initiated '4 I77 Time completed L ~

Analyst

.E$

Date analyzed

' 11.1-5. 1 III Titrant normality and multinlier determination:

Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO 3/L 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 x I00 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 50 2

.J I1.O 1

.o C-0

'40 Sooo. Vo Duplicate samwle 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)

(mg CaCO3/L)

(ml)

Ml ml ml j

IM53s qtq 40 6o*0 15S.

ZIA1 CAq lies 130 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-27 U.57

.L

'S I1111o0,03 a,.c7. C 3A7 r.I EI 10 it,11OR. 01 J,

So 31.4

.4.3 5.4 Ito.

l

0. l

.'0{*5.4 I1.

&&1

.110.

______1 0-

-\\>.

, (I')

_o S

. 13.4 1.5_

30 it 11 to. 7'*

..'64 Ca ICA.Z IM1 1

.. 4 C7.,~ I I

I 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

EnlronmantilTestilng Solutions. Inc.

Page 75 Page 3

of -4 Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO 3/L Analyst I IIII Date analyzed 51-i3.-i.

Time initiated fýq'

]

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 Laboratory control 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 to rag%)

111 40 50 1q.G t1.1 7.o)

I11.S 14 too.O%

Duplicate sample precision:

F Sample Hardness

%RPD Sample Sample ID volume Begin End Total Multiplier (mg CaCO 3/L)

((S - D) /(S+D)/21) x 100 Jinumber

.A!!

ml ml ml I10 4 Duplicate (B) 0446~ 13?.

D 10 Matrix spike recovery:

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

(mg CaCO3/L)

(ml) ml ml ml

%N65 It Io

_*0

_7._

_q.q 7.3 1q. f MOD Sample hardness (B)

Measured spike value (MV)

% R = MV I SV x 100 (mg CaCO3/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 S

o1.-,3--A i

5

3. 7.9 1

.,,.S0 itllL.0.6 DA

.CF 00%

15

.1 40'l 3.9 1 So it l10t.,:1

) ftv*

&v*l qq~ v.

4.,,

93!

II-,i"L A;0 z.6 3.1 3.A 77 nZ fl0.2 it"

-4v

'sA 77 111j1F(

1PS tWr 14J C%

%117 1 17

'1.0 174 01,0'%.,

22-1__ 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

)

Envifomental Testing Solutio Inc.

Page 7C Page 9t of q

Time initiated Time completed Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO3/L Analyst M

Date analyzed Ii.11i131 1

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 n 0.0220) -N x 1000 Laboratory control standard:

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 Duplicate sam 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 -W K SW 10k 1

tJ so 31.2 3 5A

'4.0 7

lil It A_

_L&___6(R1__.

7 I111 0t (9

.T4 s*.-

wrU'

.9 *lq75 III

.,ft 2-23.-q 37, 3.q 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

EiTS

(~Environmental Testing Solutions, Inc.

Pimephales promelas Chronic Reference Toxicant Control Chart Organism Source: Aquatox, Inc.

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

° 0.6 0.5 1.4 1.2 1.0 M 0.8 I_

0.6 0.4 I

I I

I USEPA Warning and Control Limits (75th and 90th Percentile CVs)

I I

I I

I I

i 1.4 1.2 1.0 I

I I

Laboratory Warning and Control Limits (10th and 25th Percentile CVs) oo.'.............................o..........o...................o....

0.8 0.6 0.4 jog" oxý 0&0, 01W 01%" 0VW aso os"'J' 0 00 00 *A. 0,31V 01'.0', ow"', 0ý0 %P&

PAN 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 d bV: J. Sumner 25 -A.25' A.90' Reviewed loy:

vi Kelley E. Keenan Initials: _ _"-

OETS.

l* 5 Environmental Testing Solutions, Inc.

rD Test number Test date 7-day W, CT m

(g/L KCI)

(/L KCI)

Pimephales promelas Chronic Reference Toxicant Control Chart

K (D

1 2

3 4

5 6

7 8

9 10 11 12 13 14 15 16 17 18 19 20 12-07-10 01-18-11 01-25-11 02-08-11 02-15-11 03-08-11 03-08-11 03-29-11 03-31-11 05-03-11 06-07-11 06-14-11 07-12-11 07-19-11 08-09-11 09-13-11 10-04-11 10-11-11 10-18-11 11-08-11 0.77 0.78 0.70 0.69 0.68 0.71 0.70 0.75 0.75 0.78 0.76 0.63 0.67 0.63 0.67 0.66 0.73 0.76 0.67 0.79 0.72 0.73 0.73 0.72 0.72 0.72 0.72 0.72 0.72 0.72 0.72 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71 State and USEPA S

Control Limits Sý CT-2S CT+2S 0.04 0.64 0.80 0.09 0.04 0.64 0.81 0.09 0.04 0.64 0.81 0.09 0.04 0.64 0.81 0.09 0.04 0.64 0.80 0.09 0.04 0.64 0.80 0.09 0.04 0.64 0.80 0.09 0.04 0.64 0.80 0.09 0.04 0.64 0.80 0.09 0.04 0.64 0.80 0.09 0.04 0.64 0.80 0.09 0.04 0.63 0.80 0.09 0.04 0.63 0.80 0.09 0.05 0.62 0.80 0.09 0.05 0.62 0.80 0.09 0.05 0.61 0.80 0.08 0.05 0.61 0.81 0.09 0.05 0.62 0.81 0.09 0.05 0.61 0.81 0.09 0.05 0.61 0.82 0.09 Laboratory Warning ULilts CT-S.uo CT+S, 4, 0.64 0.64 0.64 0.64 0.64 0.64 0.63 0.63 0.63 0.63 0.63 0.63 0.63 0.63 0.63 0.62 0.62 0.63 0.63 0.63 Laboratory SAs Control Lihmits CT-S O.

CT+S.

USEPA SA.7s Warning Limits CT-SLS CTO+S&7 USEPA SAAM Control Umits CTr-SLI Cr + SA&.

CV 0.81 0.82 0.81 0.81 0.81 0.81 0.81 0.81 0.80 0.80 0.81 0.80 0.80 0.80 0.80 0.79 0.80 0.80 0.80 0.80 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.57 0.57 0.57 0.57 0.57 0.57 0.57 0.57 0.57 0.57 0.57 0.56 0.56 0.56 0.56 0.56 0.56 0.56 0.56 0.56 0.88 0.27 0.88 0.28 0.88 0.28 0.87 0.27 0.88 0.27 0.87 0.27 0.87 0.27 0.87 0.27 0.87 0.27 0.87 0.27 0.87 0.27 0.86 0.27 0.87 0.27 0.86 0.27 0.86 0.27 0.86 0.27 0.86 0.27 0.86 0.27 0.86 0.27 0.86 0.27 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.44 0.44 0.44 0.44 0.44 0.44 0.44 0.44 0.44 1.00 0.33 1.00 0.33 1.00 0.33 1.00 0.33 1.00 0.33 1.00 0.33 0.99 0.32 0.99 0.32 0.99 0.32 0.99 0.32 0.99 0.32 0.99 0.32 0.99 0.32 0.98 0.32 0.98 0.32 0.98 0.32 0.98 0.32 0.99 0.32 0.98 0.32 0.98 0.32 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.39 0.39 0.39 0.39 0.39 0.39 0.39 0.39 0.39 1.05 0.06 1.06 0.06 1.05 0.06 1.05 0.06 1.05 0.06 1.05 0.06 1.04 0.05 1.04 0.05 1.04 0.05 1.04 0.05 1.04 0.05 1.04 0.06 1.04 0.06 1.03 0.06 1.03 0.07 1.03 0.07 1.03 0.07 1.03 0.07 1.03 0.07 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 IC2s 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.

$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 5th percentile 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 Variability In Whole Effluent Toxiity Applications under the National Pollutant Discharge Elimination Program. EPA-833-R-O0-003.

US Environmental Protection Agency, Cncnnati, OH.

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

Environr 1.25

"" 1.00 0.75 E- 0.50 0 b 0.25 30

'II eTS nental Testing Solutions, Inc.

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

I I

I

' l.

I I

I USEPA Acceptance Criteria (> 0.25 mg per surviving larvae)

I I

I i I

I I

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

I I

Kentucky Acceptance Limit (< 30.0%)

I i

i I

I I

i I

I I

I i

i 10 0

I I

I a.

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

20 10 0

0 I I SI 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)

"i'$n6ft9viY6, Kelley E. Keenan Initials:

Graphs generated from associated excel spreadsheet.

Excel spreadsheet ented by: J. Sumner Reviewed by:

Precision of Endpoint Measurements S

Pimephales promelas Environmental*esoudon, Inc Chronic Reference Toxicant Data Control Test number Test date Control Survival Control Mean Growth Cr CV CT MSD PMSD Cr for Control Growth

()

(m/larvae (mg/larvae) for Control Growth CV (%)

(%)

for PMSD (%)

1 2

3 4

5 6

7 8

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

Fraae 7.91 KeeK0 epend*7ve "oy Kelley E. Keenan Initials:W File: ppkclcr 11081 1.xlsx Entered by: J. Su ner Reviewed by:

Ir ES SEnv,ommn nnal Testig So talons, Inc.

Potassium Chloride Chronic Reference Toxicant Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephales promelas PpKCICR Test Number: ZL HIM Dilution preparation information:

Comments:

KCI Stock INSS number:

lt4% 1000 Stock preparation:

50 g KCI/L:

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 information for weight determination:

Transfer vessel pH =

1.60 S.U.

Date / Time in oven:

\\

IO information:

Temperature =

"4,4 0C 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, Daily feeding and renewal information:

Day Date Morning feeding Afternoon feeding Test initiation, renewal, MHSW or termination batch used Time Analvyt Time Analyst Time Analyst o

i1-0,&~tI

U O*

\\:%

O l0,t n Wadi-I I W06

>4 v'oo ItS%

il-o,4,- A 2

q-t*t j¢0

4 v-tx I2'cO tl-oq-tI g*

5 6

IIl'[

"11--,l l

~~

-s" I1"0. 11 7

uSRA" I

l I

t100 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

I ChV t(7.')a IC25 "I

IllS.-

Pange 80 of 1 0 K

Inite penae revlewy Kelley E. Keenan initals:

l SOP AT21 - Exhibit AT21.1, revision 06-01-11 V

cC I

ET l1*

a~rnme~htIIl~ng Samo.utm Inc Page 2 of 5 Species: Pimephales promelas 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 1 0]

/0 t 0 to

/0 /0

/0 1 0

/0

/010 fl 0I l /

/0

/0 to 1

2 i0

/ 0

/0 I(0

/0

/I(0 /0 C 0

'o0 '0

'0 3

0 0

60 /0 U0 to 10/0(0 I0

/0 4

/0 0 010

/

/0

/D 0

1) / i

/1 0 10 6

1 to t0 t

o1o b, /0' 1 o 1

0 t

o 9o 7

A=Paneightmg)o10 0

C) 0 II0

/Q (0 /0 00 60 t9 A yPan weight (m:g)

Tray color code::

Dal.

&I..

Analyst:

9_________

1~. 4.L1A 1.44 It.S4 l&'$9 v6. Se, I S.4.

IUz4 M..1& 14.0o i6.t.4 Date: __.____

_I B = Pan + Larvae weight (mg)

Analyst:

M4____________

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.

c7.*

1

".0

/

,'S *e.

4.'14L a rL

".1 o.0 t.do 6

o3 i

S.65 Analyst:

Weight per initial number of larvae (mg)

= C / Initial number of larvae Hand calculated.

Analyst:

A 0

'I 0D 0*

Average weight per initial Percent number of larvae (mg) reduction from control 0."t2' M%

0. 8b

-, q 7.

0.68 t

,.o7.

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 KflO E Inpender

~le~w y Kelley E. Keenan Initials:

i7W

S 0

0ETS EnvhnmantalTestln 9

In,.

Page 3 of 5 Species: Pimephales promelas PpKCICR Test Number: __A I

I I

I A = Pan weight (nmg)

Tray color code::

W.r_-

Analyst:

Date:

1o-iAG.

1.11It-I 1

'.S 411] I 4.,$

,s.C* l4.,t*

14.'st I*.

%ý A 1.,'s 1,5. 94 B = Pan + Larvae weight (nag)

A nalyst: ____1 4___1

.__1 4__1_,____

Date:

11-_1_11 I___

Iq.si* lq.4' tt.*

.'L q l6.4i

4.1c I4.,*

14.4 C = Larvae weight (mg) = B - A Handecalculated.

{,(0

-.b6

($.UlJ 6.O0 7.t

-. D#,, 3,44 2.AS 1.-11 O.PH" 0.MO Analyst:

Weight per initial number of larvae (mg)

= C /Initial number of larvae Hand calculated.

Analyst:

1W

Y r"

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

(Environmental Testing Solutions, Inc.

Pimephales promelas 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 Test dates:

November 08-15, 2011 Confi(nWL KiI]

RK t Hs hnhlida lberof Rnennml baof A-P-nam Ml.tnsa) n-* P., L.

ta.aaetiniaj-

"aW/iS* I nln M-ma)5.iht lnln Coefldimt uvdlittan W.lIedb/1nidt1nuanIra I amimihslfN Mwn 5htIhdstiA CntildaJWt*oi m

t P

r-dsdlnfoms hAnS.

wd* 5*snis A-B o-beraofhlase.(.

nansamof enn. (5.61 W5 ben.. ins nsllbeolbe I11 anabel (s)

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:

PMISD:

0.1234 15.6 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 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 _*

1ý Envi r - meta [ T em tng Solutions, Inc.

Statistical Analyses 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 ECOI 2.674 570.178 485.278 827.787 0.9 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 P. n e n4tof1 Initials:en rview bV Kelley E. Keenan Initials:

File: ppkclcr 11081 l.xlsx Entered by: J. Sunner Reviewed by:

-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 0.7-0.6-C 05-0.4:

0.3-0.2-0.1 -

0.0

-0.1

/

500 1000 1500 Dose mg/IL 0

Dose-Response Plot 1-tail, 0.05 level of significance PiNInniatln efiOB~y Kelley E. Keenan Initials:

File: ppkclcr I 1051 I.Ae Entered by: J. Sumner Reviewed by :*

'. 1. -

01) 0 0SoET 0

Pimepholes promelas Chronic Reference Toxicant Test 1" E T S Control Limits Determination Environmental Testing Solutions, Inc.

(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 2SP 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 IC25s values.

USEPA.

2000. undem~tanding and Accounting for Method Variability in Whole Effluent Todicity Applications Under the National Poflutant Discharge Elimination Program. EPA-833-R-O-003.

US Environmental Protection Agency, Cncinnati, OH.

Fie: ppk*crit083i.xlsx

,amed by.J St Renewed by-.C

ETS

1*

E..vhnlnutuI Tes g SoNlortz Inc.

Page 4 of 5 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 CaCO3/L)

Temperature (0c)

PH (SU.

___a

-+A5 190 DO (sg/.)

1;lp 7

.b

'*)

~o 450 mg KCIIL Conductivity (4*mhos/cm) 1100 I 0q0 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

750 mg KCILm(nhos/cm)

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) 1050 mg KCI/L Conductivity

([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 1M y Kelley E. Keenan Initials:

lETS Eqr41 ms E

man TibtingSolution c

n.

Species: Pimephales Promelas Page 5 of 5 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 CaCO3/L)

Hardness

\\

(mg CaCO 3/L) 0; Temperature WA (OC) pH (S.U.)

0.o5 el

.o

.00

.Q, A

Conductivity IOOO 450 ing KCIIL (ji00 10 m

Temperature C(C)

C

-usA t,.'4 2{q. 4,

,4 Q pH (S.U.)

61,03 eA",

{,*

¢I

),O f*.f

I',

600OgKCI/L 1(00 1+

'ITo-to Conductivity 600 mng KCI/L

( mo/cm) 13'0O 1*O 5"40 Q~

(iimhoslcm)

Temperature M. at (0

0) pH (S.U.)

0,0V q3/t S.ot 750 mg KCI/L Conductivity (pimhos/cm) 15l Temperature "40,

24.S *

"j..6 2.

(0c) 1...__

pH (S.U.)

10.I 4

j j13

.01 DO (mg/L>

,44*

TO o) 900 mg KCL onductivity (gnihos/cm) 100 90(3 Temperature (Oc)

_pH (S.U.)

10.1

.. 4 t

DO (mg/L).0 1050 mg KCI/L Conductivity AoD'lI Temperature zq~i~q.~

zq10

i.

214. 9

{

T-Initial Initial nitial Final I

Initial Final Initial Final m -

m -

.inm i -

~

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.

1.14 I

I I

IS d

eIv USEPA Control Limits (+/- 2 Standard Deviations)

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1.12 -

1.10 1.08 1.06 1.04 1.02 I

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USEPA Warning and Control Limits (75th and 9 0 th Percentile CVs) 0 9e e.....

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I Laboratory Warning and Control Limits (10th and 25th Percentile CVs) a...................

a.....C....*........ee..

e..........

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I 1.2 1.1 1.0 0.9 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 Ceriodaphnia reproduction for the test population.

Central Tendency (mean IC2 5)

Warning Limits (mean IC25 +/-SA-10 or SA75)

Control Limits (mean IC 25 +/- Sk25' SA.90, or 2 Standard Deviations)

Page 89 of 102 Independent review by Kelley E. Keenan initials:P, Graphs generated from associated excel spreadsheet.

Excel spreadsheet ered by: J. Sumner Reviewed by: 7

"ETS

W o.Q Io l,'Ceriod aphnia dubia ior -X Chronic Reference Toxicant Control Chart M.

(

Environmental Testing Solutions, Inc.

m State and USEPA Laboratory Laboratory a

Test number Test date 7-day ICa CT s

Control Umits S&,

Warning UmLits Ss Control Limits (g/L Naci)

(g/L NaCI)

CT-2S CT+zS CTa-S..

CT +

a'.

CT-S.

CT+Sý USEPA S.

Warning Limits CT-Sk7.

CT + S,7.

USEPA SAM-Control Limits CT-Sm CTa+ S CV 1

2 3

4 5

6 7

8 9

10 11 12 13 14 15 16 17 18 19 20 07-13-10 08-03-10 09-14-10 10-05-10 10-05-10 11-02-10 12-07-10 01-18-11 02-08-11 03-08-11 04-05-11 04-05-11 05-03-11 06-07-11 07-12-11 08-09-11 09-13-11 10-04-11 10-11-11 11-08-11 1.07 1.06 1.07 1.08 1.08 1.09 1.06 1.06 1.06 1.05 1.06 1.07 1.07 1.05 1.06 1.06 1.06 1.07 1.06 1.05 1.07 0.01 1.05 1.09 0.09 0.99 1.07 0.01 1.05 1.09 0.09 0.99 1.07 0.01 1.05 1.09 0.09 0.99 1.07 0.01 1.05 1.09 0.09 0.99 1.07 0.01 1.05 1.10 0.09 0.99 1.07 0.01 1.05 1.10 0.09 0.99 1.07 0.01 1.05 1.10 0.09 0.99 1.07 0.01 1.05 1.10 0.09 0.99 1.07 0.01 1.05 1.09 0.09 0.99 1.07 0.01 1.05 1.10 0.09 0.99 1.07 0.01 1.05 1.09 0.09 0.98 1.07 0.01 1.05 1.09 0.09 0.98 1.07 0.01 1.05 1.09 0.09 0.98 1.07 0.01 1.05 1.09 0.09 0.98 1.07 0.01 1.04 1.09 0.09 0.98 1.07 0.01 1.04 1.09 0.09 0.98 1.07 0.01 1.04,

1.09 0.09 0.98 1.07 0.01 1.04,

1.09 0.09 0.98 1.07 0.01 1.05 1.08 0.09 0.98 1.07 0.01 1.05 1.08 0.09 0.98 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.16 0.18 0.89 1.15 0.18 0.89 1.15 0.18 0.89 1.15 0.18 0.89 1.15 0.18 0.89 1.15 0.18 0.88 1.15 0.18 0.88 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.26 0.48 0.59 1.26 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.25 0.48 0.59 1.55 0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.41 1.56 0.67 0.41 1.56 0.67 0.41 1.56 0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.41 1.55 0.66 OA1 1.55 0.66 0.41 1.55 0.66 0.41 1.55

.0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.41 1.55 0.66 0.40 1.54 0.66 0.40 1.74 0.01 1.74 0.01 1.73 0.01 1.73 0.01 1.74 0.01 1.74 0.01 1.74 0.01 1.74 0.01 1.74 0.01 1.74 0.01 1.73 0.01 1.73 0.01 1.73 0.01 1.73 0.01 1.73 0.01 1.73 0.01 1.73 0.01 1.73 0.01 1.73 0.01 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 IC25 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.

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 IC25 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.

File: CdNaCICR 110811.xAsx Entered by'. J..Wpmner Reviewed by&__

0-ETS 5

Environmental Testing Solutions, Inc.

Ceriodaphnia dubia Chronic Reference Toxicant Control Chart Precision of Endpoint Measurements 0

0 Lu Lu 0U T

40 35 30 25 20 15

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Minimum Acceptance Criteria (> 15.0 offspring per surviving female)

I i

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40

'~.2 30 20 10 North Carolina Acceptance Limit (< 40.0%)

Kentucky Acceptance Limit (< 30.0%)

50 40 30 20 10 n

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I I I USEPA Upper PMSD Bound (90'h percentile < 47.0%)

S.

I I I

I.......

I I

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)

Pa oe.91 of 102 enlendent evew by Kelley E. Keenan Initials:

Pr Graphs generated from associated excel spreadsheet.

Excel spreadsheet entered by: J. Sumner Reviewed by:

N

.ETS Environmental Testing Solutions, Inc.

Precision of Endpoint Measurements Ceriodaphnia dubia Chronic Reference Toxicant Data Test Test Test date number Control Survival Control Mean Reproduction

(%)

(offspring/female) 1 2

3 4

5 6

7 8

9 10 i1 12 13 14 15 16 17 18 19 20 07-13-10 08-03-10 09-14-10 10-05-10 10-05-10 11-02-10 12-07-10 01-18-11

.02-08-11 03-08-11 04-05-11 04-05-11 05-03-11 06-07-11 07-12-11 08-09-11 09-13-11 10-04-11 10-11-11 11-08-11 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 31.6 28.2 32.9 30.2 30.8 30.5 32.2 31.9 30.7 32.6 32.8 33.0 30.9 28.9 32.2 28.3 32.1 32.8 31.3 29.0 cr for Control Mean Reproduction (offspring/female) 29.9 30.9 30.7 30.7 30.7 30.9 31.0 31.0 31.2 31.3 31.5 31.4 31.2 31.3 31.1 31.2 31.3 31.3 31.1 CV CT for Control

(%)

Reproduction CV (%)

MSD PMSD

(%)

for PMSD (%)

CT 6.6

.4.8 4.7 8.7 7.6 4.0 4.9 4.8 6.3 6.9 4.8 5.9 4.5

.6.7 6.0 5.0 8.2 6.1 3.0 7.6 5.7 5.3 6.2 6.5 6.0 5.9 5.8 5.8 5.9 5.8 5.8 5.7 5.8 5.8 5.8 5.9 5.9 5.8 5.9 2.4 2.3 1.8 3.0 2.0 2.2 2.0 2.0 2.3 1.8 2.0 2.2 1.9 2.5 2.0 1.9 1.9 2.1 2.1 1.8 7.6 8.0 5.3 10.0 6.5 7.3 6.2 6.3 7.4 5.4 6.0 6.7 6.1 8.7 6.2 6.8 5.8 6.5 6.6 6.1 7.8 7.0 737 7.5 7.5 7.3 7.2 7.2 7.0 6.9 6.9 6.8 7.0 6.9 6.9 6.8 6.8 6.8 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.

P.AQ e2tof1 enntreiew y Kelley E. Keenan Initials:

File: CdNaCICR_11081.xlsx Entered by: J. Sugner Reviewed by:

S 6

00*ETS Page 1 of 6 Sodium Chloride Chronic Reference Toxicant Test (EPA-821-R-02-013 Method 1002.0)

Species: Ceriodaphnia dubia CdNaCICR #:

t2?'l Dilutionpreparation information:

Comments:

NaCI Stock INSS number:

I W 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.U IVA 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 6

I1-,-

clb ItO--tl'0 5

11- %-!rI

o.

Control information:

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-/j CV:"

< 40.0 %

ICs, Pew.

of 102

-In oenae*tjevlew By Kelley E. Keenan Initials:

=

SOP AT14 - Exhibit AT14.1, revision 06-01-11

J aETS Page 2 of 6 Species: Ceriodaphnia dubia CONTROL CdNaCICR #:

12t1 Survival and Reproduction Data I:

Replicate number Day 1

2 3

4 5

6 7

8 9

10 I

Young produced O

0) 0 i0 0

0UO 0

Adult mortality 1 '

- I 1

2 Young produced C) 60 Adult mortality JL

%. HI

'% + '-, I_

I

,*_T[

=

3 Young produced

__0 a

c5 T0~2 Adult mortality L I'-

lL-

'1 1

4 Young produced~

5 1L i

f-+/-TA -L Adult mortality I'--"

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k

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6oa Young produced 0..

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0

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'20 c.)

Adult mortaity I

I_ \\.

I***

\\.

U 1

\\-

I

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6 TYoung produced [

I i 0 01 10 2 To]a young produced 2A zJ a-

-Z

-4 LI Final Adult Mortality L -C-

=_

C C :-

L Z.

Tafor 3ou Broods 4

y 7 ý t ".

'?

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:

W 0.

Mean.Offsprinemale:

".,0 600 mg NaCiJL Survival and Reproduction Data 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

A d u lt m o rta lity L_

_L--

3 Young produced

0)

Q 0

-1 6

Adult mortality I

01 c1 L II 3

Young produced

%A Adult mortality I

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1+

5 Young produced T

0% Q_

to LA 1

12 11 I C1I It.

Adult mortality

%.-I C

C Ij~

~

jL 1%_

6 You. prdcd0 1Q

(

i

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'-IL

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Young produce 0

ý.I o

.(

Total young produced j

3o

.3

-n 3'L

'Z

'A z Final Adult Mortality I_

I.

I

\\.-

\\ý 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 SOP AT14 - Exhibit AT14.1, revision 06-01-11 Page 94 of 102 Ind-epen gt review by Kelley E. Keenan Initials____

ETS Page 3 of 6 Species: Ceriodaphnia dubia 800 mg NaCI/L CdNaCICR #:

171 Survival and Reproduction Data 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 mortalit Adult mortality t__

%,- I

\\,- I_

I_ \\

V-2 Young produced 1C1 I

C)~

I

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3 Young prod uced 0

0 CT0 cc)

Adult mortality

_I

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4 Young produced

%j 1 %A_

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Young produe toIJII.

2.I I'

~..

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in.1i Adult mortality

]___

61 Young produced C)

' V_.

1 0101 01010 Adult mortality 7

Young produced

%4 1 Total young produced

-3 1 2A n

z 2.

30 7_j k Final Adult Mortality J'

-- I Note: Adult mortality (L = live, D = dead), SB = split brood (single broqd split betwedn two days), CO = carry over (offspring 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 and Reproduction Data Replicate number Day 1

2 3

4 5

6 7

1 8

9 10 I

Young produced F

Q 0

0 1

01__"

! Adult mortality

\\

L I

t 4

t-- I-=-

2 Young produced" 6 1 L0I

.Il II, Adult mortality L-- I-I L

"I L-3 Young produced f 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

SOP ATI4 - Exhibit AT14.1, revision 06-01-11 I

Pa e 95 of 102 Indapendetrview by Kelley E. Keenan Initials:

1C-

0ETS Z ErMMW 1ý~Wcmmnn1lSu Page 4 of 6 Species: Ceriodaphnia dubia 1200 mg NaCI/L CdNaCICR #:

211-1 Survival and Reproduction Data Replicate number Day 1

2 3

4 5

6 7

8 9

10 Adult mortality f j.

\\

L.-

2 Young produced C0 Adult mortalityI 1--

I --

" I I" "

3 1 Young producedI CI)I I

e.

I Adult mortality L. j

1. -

'\\

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:

"12.1 1.

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 l)

Adult mortality k-1..-

J L..

\\---

\\_

-I 2

Youngproduced 1.,,i o I 0 IL i

IrF1 0 10 3_J Young produced Adult mrtality

__1

__ 0__

4 Young produced 0

f

[

1 0 Adult mortality k-

,.1~-I L

J_

-JL--

I I E%...-_

5 Young produced C* I 6

Yugproduced 7 Young produced 0

0,-

C

~

~.Q.

Total young produced o

o i

0 6

Final Adult Mortality t

X.-

W T.

X?

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.

SOP AT14 - Exhibit AT14.1, revision 06-01-11 Se 96 of 10 Ini*L~en r:eview 2yKelley E.Keenan Initials:

V____

i* CDC O:ETS t

)

(51 Environmental Testing Solutions, Inc.

Control Verification of Ceriodaphnia Reproduction Totals i5.

m m

0 C,

Re plicate number Total Day 2

3 4

5 6

7 8

9 10 1

0 0

0 1 0.

0 2

0 0

0 3

0 0

0 4

5 5

4 4

3 4

4 4

41 5

9 9

13 10 10 11 12 9

10 10 103 6

0 0

0 7

15 13 16 14 14 14 16 13 16 15 146 Total 29 27 33 28 27 29 32 26 30 29 290 600 mg NaCI/L Re licate number Day 1

2 3

4 1 5 1 L.

7 8

9 10 I

0 0

0 2

0 0

0 3

0 0

0 4

4 5

4 4

5 4

4 4

4 3

41 5

10 9

12 10 10 11 12 12 10 9

105 6

0 0

0 7

15 15 14 15 16 14 16 13 15 13 146 Total 29 29 30 29 31 29 32 29 29 25 292 800 mg NaCI/L Re plicate number Total Day 1

2 3

4 5

6 7

8 9

10 1

0 0

0 2

0 0

0 3

0 0

0 4

4 3

4 4

5 5

4 4

41 5

11 10 11 12 11 10 10 10 10 9

104 6

0 0

0 7

15 14 14 14 13 IF 15 13 16 15 144 Total 30 27 29 30 29 30 29 27 30j28 289 1000 mg NaCMIL Re plicate number Total Day 2

3 1 4 5

6 7

8 9

10 1

0 0

0 10 0

2 0

0 0

3 0

0 0

4 3

3 4

4 4

4 37 5

9 9

11 10 10 9

11 10 10 10 99 6

0 0

0 7

12 14 13 13 15 12 14 14 14 13 134 Total 24 27 27 26 29 25 29 28 28 27 270 1200 mg NaCI/L Re licate number Tota Day 1

2 3

4 5

6 7

8 10 Total 1

0 0

0 2

0 0

-0 0

0 0

0 3

0 0

0 4

3 2

0 0

0 3

0 2

0 0

10 5

0 0

4 1

4 0

3 1

2 2

17 6

2 0

0 0

0 5

0 0

7 7

6 5

5 4

7 0

6 5

3 6

47 Total 11 7

9 5

11 8

9 885 8

81 1400 mg NaCI/L Da0 0mg a

Replicate number Total Day 2

3 14 5

6 7

8 9

10 1

0 0

0 2

0 0

0 3

0 0

0 4

0 0

1 0

0 0

0 5

0 0

0 2

0 0

2 6

0 0

0 7

0 2

0 0

3 0

0 5

Total 0

2 0

2 1

0 0

3 0

0 8

File: CdNaCICR_110811.xlsx Entered by: J. Surpner Reviewed by:

ur M0ET 7amr 4

n io m na etn oui nI c

Ceriodaphnia dubia Chronic Reference Toxicant Test EPA-821-R-02-013, Method 1002.0 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Test number:

Test dates:

CdNaCICR #127 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:

PMSD:

1.766 6.1 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 (10d' percentile) = 13%.

Upper PMSD bound determined by USEPA (90* 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:

SETS

!1 EnvronmentNi stsngSoluthflim Statistical Analyses 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 Conc-mg/L Mean N-Mean Mean Min Max CV%

N D-Control 29.000 1.0000 29.000 26.000 33.000 7.624 10 600 29.200 1.0069 29.200 25.000 32.000 6.211 10 800 28.900 0.9966 28.900 27.000 30.000 4.143 10

1000 27.000 0.9310 27.000 24.000 29.000 6.048 10

1200 8.100 0.2793 8.100 5.000 11.000 25.667 10

.1400 0.800 0.0276 0.800 0.000 3.000 141.912 10 1-Tallea Isotonic t-Stat Critical MSD Mean N-Mean 29.100 1.0000

-0.259 2.287 1.766 29.100 1.0000 0.129 2.2l7 1.766 28.900 0.9931 2.590 2.287 1.766 27.000 0.9278 27.065 2.287 1.766 8.100 0.2784 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

IC40

. 1100.95 5.21302 1090.81 1110.54 0.0871 0.8-IC50 1131.75 5.41741.1121.38 1141.94 0.1709 0.7:

0.6 C00.5-0.41 I.3-0 S00 1000

.Dose mg/L 1500 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: ______

Ceriodaphnia dubia Chronic Reference Toxicant Test D

WControl Limits Determination v1

(

Environmental Testing Solutions, Inc.

m State and USEPA Laboratory Laboratory Test number Test date 7-day IC, CT S

Control Limits SA Warning Limits 5

L&M Control Umits S&",

S(g/L NaCI)

(g/L NaCI)

Cr-ns ar+2s r

C+ cr-sj s~

-sL c n+ s,,.

USEPA Warning Limits CT-S,.

CT + S..

USEPA So Control Limits a-CT.SA CT + SAM.

1 2

3 4

5 6

7 8

9 10 11 12 13 14 15 16 17 18 19 20 07-13-10 08-03-10 09-14-10 10-05-10 10-05-10 11-02-10 12-07-10 01-18-11 02-08-11 03-08-11 04-05-11 04-05-11 05-03-11 06-07-11 07-12-11 08-09-11 09-13-11 10-04-11 10-11-11 11-08-11 1.07 1.06 1.07 1.08 1.08 1.09 1.06 1.06 1.06 1.05 1.06 1.07 1.07 1.05 1.06 1.06

1.06 1.07 1.06 1.05 1.06 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07.

1.07 1.07 1.07 1.07 0.01 1.05 1.08 0.09 0.01 1.05 1.08 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.10 0.09 0.01

.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.04 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.09 0.09 0.01 1.05 1.08 0.09 0.98 0.98 0.98 0.99 0.99 0.99 0.99 0.99 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 1.15 1.15 1.15 1.16 1.16 1.16 1.16 1.16 1.15 1.15 1.15 1.15 1.15 1.15 1.15 1.15 1.15 1.15 1.15 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.88 0.88 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.88 0.88 0.88 0.88 1.24 1.25 1.25 1.25 1.26 1.26 1.25 1.25 1.25 1.25 1.25 1.25 1.25 1.25 1.25 1.25 1.25 1.25 1.25 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 1.54 1.55 1.55 1.55 1.56 1.56 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.54 0.66 0.66 0.66 0.66 0.67 0.67 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.40 0.42 0.41 0.41 0.41 0.41 0.41 0.41 0.41 0.41 0.41 0.41 0.41 0.41 0.41 OA1 OA1 0.40 0.40 1.72 1.73 1.73 1.74 1.74 1.74 1.74 1.74 1.73 1.73 1.73 1.73 1.73 1.73 1.73 1.73 1.73 1.73 1.73 CV 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 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 7 5e 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: Ceriodaphnia dubia CdNaCICR #:

M.1 Daily Chemistry:

_ _[_

_Day

_0 1

2 Concentration Parameter pH(..

t*

0S I-A 1 9900 Conductivity (gmhos/cm) t l

CONTROL Alkalinity (mg CaCO3/L) w4.1 Hardness (Mg CaC0 3/L) c~

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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

~

L 1200 mg NaCVL Conductivity L.O Temperature 260~*

____I___

(0 c) z~

DH (S.U.)

DO (mg/L) 1400 mg NaCI/L Conductivity (jimhos/cm)

I :bo II

+-0f 1.6. 0 qq [ -.

Temperature (0C)

STOCK IConductivity I (Amhos/cm) 1 10.5=

Initial 14 itFinal I Initial 11 Final I Initial 11 Final

,'q.9 w.*

SOP AT14 - Exhibit AT14.1, revision 06-01-11 P

e 01 ofIn ewyelley E. Keenan Initials:W

I II II I

I I

rn

ETS Page 6 of 6 Species: Ceriodaphnia dubia 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 pH (S.U.)

10AM80 3O ck

.,L

%It

%.c.

L.

.s\\

x**

600 mg NaCI/L Conductivity iý"30 (5eo (gmhos/cm)

.75(1 Temperature

-4.0 2LSZ IS..

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pHo(S.U.)

Pro

.15o 800 mg NaCl/L Conductivity (1IChosncm)

L* 00 so Temperature V

T*.-

24

_o__

__IN__

PH (S.U.)

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.09L.

13-03 1,04_____

1000~~D mg aCIL dutiit 1000 mg NaCIIL Conductivity (jimhos/cm)

LOS

',1 oO Temperature 7--4 a

{

1400 mg NaCI/L Conductivity

-0 Temperature "2.qh

-j.

2q.1-S 7

2.4.

"-S A

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Initial 11 Final I Initial II Final I Initial II Final I Initial II Final I lb lb ii lb -

lb -

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PERMII-IEE NAMEIADDRESS (Include Facility Name/Location if Different)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 (INTEROFFICE OPS-5N-SSQ.

SODDY - DAISY, TN 37384 Facility TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

FMJOR Form DISCHARGE MONITORING REPORT (DMRJ (SUBR 01)

OMB I

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30/30 RCORDR TREATMENT PLANT MEASUREMENT 50050 1

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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 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 1

I CODE LUMMEIN I EXPL UA NAI'I.

I IUF Ut-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.

~PA FOrE J~ZU-1 (REV 3153)

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

Previous editions may be used Page 1 of 1

PERMIIT-EE NAMEIADDRESS (Include Facility Name/Location if Different)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 j

E NTEROFFICE OPS-5N-SQN).

SODDY - DAISY TN.337384 Facii TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

MAJOR DISCHARGE MONITORING REPORT (DMR)

(SUBR 01)

NI TN026450 110 G I F-FINAL PERMIT NUMBER NDISCHARGENUMBERI RECYCLED COOLING WATER MONITORING PERIOD EFFLUENT Form Approved.

OMB No. 2040-0004 I YEARI MO IDAYI Froml -Il 1 11 01 I YEA R

AY I NO DISCHARGE IX1 M 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

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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

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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 Facility Name/Location if Different)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000

- (INTEROFFICE OPS-5N-SQ.N)-

SODDY - DAISY TN 37384 Fa.cility TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR DISCHARGE MONITORING REPORT (DMRJ (SUBR 01)

TN026450 I

110 T F -FINAL PERMIT NUMBER DISCHARGE NUMBER RECYCLED COOLING WATER MONITORING PERIOD

]EFFLUENT Form Approved.

OMB No. 2040-0004 I YEAR M

DAY I

YEARI MO I DAY From l 1111 101I To IlI 11 30 NO DISCHARGE 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

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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 TpTELEPHONE 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 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)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000

..INTEROFFICE OPS-5N-SQN)

SODDY - DAISY1N._373 84 Facilty.

TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

MAJOR DISCHARGE MONITORING REPORT (DMAfRJ (SUBR 01)

TN0026450 118 G F - FINAL PERMIT NUMBER DISCHARGE NUMBER WASTEWATER & STORM WATER MONITORING PERIOD EFFLUENT Form Approved.

OMB No. 2040-0004 I YEARI MO.

DAY I YEARI MO DA 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*

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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 persons directly responsible for gathering the StViePresident 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, 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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