Text

NPDES Permit No. TN0026450 - Discharge Monitoring Report (DMR) for September 2012 and Revised DMR for August 2012
	ML12286A328
Person / Time
Site:	Sequoyah
Issue date:	10/10/2012
From:	John Carlin; Tennessee Valley Authority
To:	Morgan C; Office of Nuclear Reactor Regulation, State of TN, Dept of Environment & Conservation, State of TN, Div of Water Pollution Control
References
	TN0026450
	Download: ML12286A328 (108)
	v • d • e

Tennessee Valley Authority, Post Office Box 2000, Soddy Daisy, Tennessee 37384-2000 October 10, 2012 Ms. Christina Morgan Tennessee Department of Environment and Conservation Division of Water Pollution Control Enforcement & Compliance Section 6 th Floor, L & C Annex 401 Church Street Nashville, Tennessee 37219

Dear Ms. Morgan:

TENNESSEE VALLEY AUTHORITY (TVA) - SEQUOYAH NUCLEAR PLANT (SQN) - NPDES PERMIT NO. TN0026450 - DISCHARGE MONITORING REPORT (DMR) FOR SEPTEMBER 20"12 AND REVISED DMR FOR AUGUST 2012 Enclosed is the September 2012 Discharge Monitoring Report for Sequoyah Nuclear Plant. The toxicity report and revision to Discharge Number 101T, Biomonitoring for Outfall 101 for August 2012, are also attached. There were no exceedances during the monitoring period. If you have any questions or need additional information, please contact Brad Love by email at bmlove@tva.gov or by phone at (423) 843-6714.

I certify under penalty of law that this document and all attachments were prepared under my direction or supervision in accordance with a system designed to assure that qualified personnel 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.

Sequoyah Nuclear Plant Enclosures cc (Enclosures):

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

PERMIIrEE NAME/ADDRESS (Include Facility Name/Location if Differentj-Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 I[NTEROFFICE OPS-5N-SQ.N SODDY - DAISY, TN 37384 Facilj.:, TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR DISCHARGE MONITORING REPORT (DMR)

(SUBR 01)

TN0026450 t101 G

F-FINAL PERMIT BEDISCHARGE NUMBE DIFFUSER DISCHARGE MONITORING PERIOD EFFLUENT Form Approved.

OMB No. 2040-0004 F

r YARm I Mr DAY I T EAo l MO I DAY From 1 2-- 0 0

To 1 12 109 1301 NO DIS iSCHARGE NOTE: Read instructions before completing this form.

PARAMETER FREQUENCY SAMPLE OF TYPE QUANTITY OR LOADING QUALITY OR CONCENTRATION NO.

EX FREQUENCY OF ANALYSIS SAMPLE TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS TEMPERATURE, WATER DEG.

SAMPLE 41.3 0

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SAMPLE 29.1 04 0

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CONTI CALCTD EFFLUENT GROSS REQUIREMENT DAILY MX CIHR NUOUS SAMPLE MEASUREMENT PERMIT REQUIREMENT NAM1_J'rITILE 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. Cadin properly gather and evaluate the information submitted. Based on my inquiry of the person or

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OFFICER OR AUTHORIZED AGENT AREA NUMBER MO DAY TYPED OR PRINTED I

CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

No closed mode operation. Veliger monitoring data is included as an attachment. The following injections occurred: 1. Floguard MS6236 (max. calc. conc. was 0.057mg/L-limit 0.2mgIL) 2.

Biodetergent 73551 (max. calc. conc. was 0.038mg/L-limit 2.Omg/L) 3. Spectrus CT1300 (max. calc. conc. was 0.037mg/L-Iimit 0.050mg/L)

EPA Form 3320-1 (REV 3199)

Pirevious editions may be used Page I of I

Mean # of Water Mean# of Water NOTES: %

Sample Date

% Settlers Sample Date Asiatic Temp. ('C)

LOCATION SUB LOCATION Gravid Asiatic COLLECTED BY CClamsm3 TClam 01/03/2012 14 100 26 01/03/2012 0

26 1-25-545 PKS 01/10/2012 01/17/2011 01/24/2012 01/31/2012 02/07/2012 02/14/2012 02/21/2012 02/28/2012 03/06/2012 03/13/2012 03/20/2012 03/27/2012 04/03/2012 04/10/2012 04/18/2012 May 2012 June 2012 July 2012 August 2012 September 2012 0

0 0

1623 229 79 326 0

0 0

1.3 0

20 5

9 10 13 17.6 12 8.3 26.5 11.1 11.7 13 14.6 17.2 18 22 18.8 01/10/2011 01/17/2011 01/24/2012 01/31/2012 02/07/2012 02/14/2012 02/21/2012 02/28/2011 03/06/2012 03/13/2012 03/20/2012 03/27/2012 04/03/2012 04/10/2012 04/18/2012 0

0 0

9 10 13 17.6 12 8.3 26.5 11.1 11.7 13 14.6 17.2 18 22 18.8 RCW 1-ISV-24-1234 1-25-545 1-25-545 1-25-545 1-24-1234 1-25-545 1-ISV-24-1234 1-ISV-24-1234 1-ISV-24-1234 1-ISV-24-1234 1-ISV-24-1234 1-ISV-24-1234 1 -ISV-24-1234 1-1SV-24-1234 WBE PB WDT CR BB WE CR WBE WBE WBE WBE WBE PB PB MJW No Samples Collected No Samples Collected No Samples Collected No Samples Collected No Samples Collected

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

N+/-ame TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 (INTEROFCE OPS-5N-SQN.)

SODDY - DAISY TN.37384 Facil_ y_

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

MAJOR For DISCHARGE MONITORING REPORT (DMR)

(SUBR 01)

OM TN0026450 F-FINAL PERMIT NUMBER DISCHARGE NUMBER BIOMONITORING FOR OUTFALL 101 MONITORING PERIOD EFFLUENT rm Approved.

BI No. 2040-0004 Froml 1 109011 Tol11 09L L 30 NO DISCHARGE liZ ]

NOTE: Read instructions before completing this form.

PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO.

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TELEPHONE DATE direction or superyision 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 President 43 8.-0!

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4.3 land complete. I am aware that there are significant penalties for submitting false information, SIGhTJp:F PRINCIPAL EXECUTIVE T

including the possibility of fine and imprisonment for knowing violations.

OFFICeR'OR AUTHORIZED AGENT AREA NUMBER MO DAY

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CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

Toxicity was not sampled in September 2012.

EPA Forim 3320-1 (REV 3199)

Previous editions may be used Page 1 of 1

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

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

.f.-.NTEROFFICE OPS-5N-SQON)

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

MAJOR Form Approved.

DISCHARGE MONITORING REPORT (DMR)

(SUBR 01) 0MB No. 2040-0004 TN024 1103 G

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423 843-7001 12 10 10 S

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PRINCIPAL EXECUTIVE including the possibility of fine and imprisonment for knowing violations.

SI ICE -OR AUTHORIZED AGENT AREA NUMBER TYPED OR PRINTED I CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all aftachments here)

EPA Form, 3320-1 (REV 3I9S)

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

Previous editions may be used Page 1 of 1

PERMII-TEE NAME/ADDRESS (Include Facility Name/Location if Different)

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(SUBR 01)

TN0026450 110 G F - FINAL PERMIT NUMBER DISCHARGE NUMBER RECYCLED COOLING WATER MONITORING PERIOD

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DEG C CONTIN CALCTD EFFLUENT GROSS VALUE REQUIREMENT DAILY MX uous 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 dlirection or supervision in accordance with a system designed to assure that qualified personne John T. Carlin property 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 Seiuovah Site Vice President nformation, the information submitted is, to the best of my knowledge and belief, true, accurate, E_

423 843-7001 12 PN PAn and amp;.;

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O OR AUTHORIZED AGENT AREA NUMBER TY OR PRINTED I 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

PERMIIrEE NAMEIADDRESS (Include Facility Name/Location if Diffement)

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

--- LNTEROFFICE OPS-5N-SQ_.N)

SODDY - DAISY. TN 37384 Faciljiy TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON co UNTY.-

ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

DISCHARGE MONITORING REPORT (DMR)

TN0026450 11 PERMIT NUMBER DISCHARGE R

MONITORING PERIOD YEAR MO I DAY I YEAR MO DAY From J12 09 101 Tol 121091301 MAJOR (SUBR 01)

F - FINAL RECYCLED COOLING WATER EFFLUENT Form Approved.

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PARAMETER QUANTITY OR LOADING I

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No Discharge this Period EPA Form 3320-1 (REV 3199)

Previous editions may be used Page I of I

PERMI-rEE NAME/ADDRESS (Include Facility NamelLocation if Different)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 (NTEROFFICE OPS-5N-SQN).

SODDY - DAISY TN 37384 Facilijy TVA - SEQUOYAH NUCLEAR PLANT Location HAMILT.ON COUNTY ATTN: Brad Love NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

DISCHARGE MONITORING REPORT (DMRJ TNO026450 118 G PERMIT NUMBER DISCHARGE NUMBERJ MONITORING PERIOD SYEAR I MO I DAY T o A

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(SUBR 01)

OMB No. 2040-0 F - FINAL WASTEWATER & STORM WATER EFFLUENT NO DISCHARGE

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004 PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO.

FREQUENCY SAMPLE EX OF TYPE AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS OXYGEN, DISSOLVED (DO)

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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 sident information, the information submitted is, to the best of my knowledge and belief, true, accurate.

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NUMBER YEAR MO DAY TYPED OR PRINTED CO COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all aftachments 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 Page 1 of I

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

N+/-ame TVA - SEQUOYAH NUCLEAR PLANT Adddress P.O. BOX 2000

-JLNTEROFFCE OPS-5N-SQN)

SODDY - DAISY, TN_373_84 Fac.lli' TVA - SEQUOYAH NUCLEAR PLANT L

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MONITORING PERIOD

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NOTE: Read Instructions before completing this form.

004 PARAMETER QUANTITY OR LOADING QUALITY OR CONCENTRATION NO.

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TELEPHONE DATE d-irection 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 ice President 4 %

information, the information submitted is, to the best of my knowledge and belief, true, accurate, 423 843-700i 1 i.

Sequoyah Site Vice President and complete. lam aware that there e sign..n"t fcf Subrinting iai-, informajion, SIGNAJIE OF PRINCIPAL EXECUTIVE I

including the possibility of fine and imprisonment for knowing violations.

OFFICER OR AUTHORIZED AGENT AREA NUMBER MO DAY TYPED OR PRINTED CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

Toxicity sampling began on August 12 and ended on August 17. This is the revised August 2012 DMR data set for 101T. The toxicity report is attached.

EPA Form 3320-1 (REV 3199)

Pmvious editions maybe used Page 1 of I

TENNESSEE VALLEY AUTHORITY TOXICITY TEST REPORT INTRODUCTION / EXECUTIVE

SUMMARY

Report Date: September 17, 2012

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: 3,.491

8. Outfall Tested: 101

9. Dates Sampled: August 12-17, 2012

10. Average Flow on Days Sampled (MGD): 1741.838, 1748.488, 1747.571

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

12. Test Dates: August 14 - 21, 2012

13. Test Type: Short-term Chronic Definitive

14. Test Species: Fathead Minnows (Pimrehales 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: IC,5 = 43.2%

17. Test Results: Outfall 101: Pitnephales promelas: IC 25 > 100%

Ceriodaphnia dubia: IC25 > 100%

UV treated Outfall 101: Pimephales promelas: IC, 5 > 100%

Page 1 of 99

18. Facility

Contact:

Brad Love Phone #: (423) 843-6714

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

20. Lab

Contact:

Jim Sumner Phone #: (828) 350-9364

21. TVA

Contact:

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

22. Notes: Outfall 101 samples collected August 12 - 17, 2012, 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 2 of 99

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 08-12-12 0645 to 08-13-12 1540 0.8, 0.7t

<0.10 08-14-12 1213 08-13-12 0545 08-15-12 1115 08-12-12 0705 to 08-14-12 1213 Intake 08-13-12 0605 08-13-12 1540 0.9

<0.10 08-13-12 0605 08-15-12 1115 101 08-14-12 0700 to 08-15-12 1553 1.9 1.9t

<0.10 08-16-12 1114 08-14-12 0700to 08-16-12 1114 Intake 08-15-12 0620 08-15-12 1553 2.0

<0.10 08-16-12 1114 08-16-12 00to 08-18-12 1112 101 08-17-12 0600 08-17-12 1338 2.2, 2.2 t

<0.10 08-19-12 1115 08-17-12 0600 08-20-12 1114 08-16-12 0710to 08-18-12 1127 Intake 08-16-12 0715 08-17-12 1338 2.4

<0.10 08-19-12 1115 08-17-12 0615 08-20-12 1114

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

4. Sample Manipulation: Samples from Outfall 10 1 and intake were warmed to test temperature (25.0 +/- 1.0°C) 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 99

Pimephales promelas Test Organisms:

Ceriodaphnia dubia In-house Cultures

1. Source:

Aquatox, Inc.

2. Age:

20 - 20.22 hours2.546296e-4 days 0.00611 hours 3.637566e-5 weeks 8.371e-6 months old

< 24-hours old Test Method Summary:

1. Test Conditions:

Static, Renewal Static, Renewal

2. Test Duration:

7 days Until at least 60% of control females have 3 broods 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 I

08-14-12 1200 ET 08-14-12 1213 ET 08-21-12 1106 ET 08-21-12 1122ET Mean = 24.7°C (24.3 - 25.1 0C) 08-14-12 1148 ET 08-21-12 1053 ET Mean = 24.9°C (24.6 - 25.30C)

Test Temperature: UV-Treated Outfall 10 1:

Mean = 24.8°C (24.2 - 25.1 0C)

9. Physical / Chemic Measurements:

al 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.

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

10. Statistics:

Page 4 of 99

TOXICITY TEST RESULTS (see Appendix C for Bench Sheets)

1.

Results of a Pimephales promelas Chronic/ 7-day Toxicity Test.

(Genus species)

(Type / Duration)

Conducted August 14.- 21, 2012 using effluent from Outfall 101.

Test Percent Surviving Solutions (time interval used - days)

(% Effluent) 1 2

3 1 4

[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%

100 100 100 100 100 100 100 100.0%

100 100 100 100 100 100 100 Intake 100 100 100 100 100 100 95 Mean Dry Weight (mg)

Test Solutions (replicate number)

(%1Effluent) 1 2

J 3

4 I

Mean Control 0.583 0.615 0.592 0.544 0.584 10.8%

0.535 0.669 0.686 0.649 0.635 21.6%

0.608 0.559 0.626 0.686 0.620 43.2%

0.582 0.563 0.487 0.614 0.562 86.4%

0.614 0.555 0.563 0.690 0.606 100.0%

0.532 0.673 0.506 0.571 0.571 Intake 0.537 0.569 0.551 0.687 0.586 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/LC50: TUc = 100/IC25 Page 5 of 99

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 August 14 - 21, 2012 using effluent from Outfall 101.

Percent Surviving Test (time interval used - days)

Solutions

(% Effluent) 6 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 Test Solutions Reproduction (#young/female/7 days)

Test Soluetis Data (replicate number)

(% Effluent) 1 2 J3 ] 4 I 5 6 [ 7 1 8 [ 9 110 [Mean Control 30 33 29 33 31 28 34 32 30 32 31.2 10.8%

36 34 33 33 35 35 34 34 31 32 33.7 21.6%

36 31 33 31 30 37 35 33 32 30 32.8 43.2%

37 34 34 31 31 31 37 34 34 35 33.8 86.4%

34 34 36 34 35 33 34 35 36 37 34.8 100.0%

38 37 37 34 32 38 35 39 37 38 36.5 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 = 1 00/LC5 0: TUc = 100/IC25 Page 6 of 99

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 August 14 - 21, 2012 using water from Intake Percent Surviving Test (time interval used - days)

Solutions 1

1 5

61 1(%

Effluent) 1 2

13 4

5 Control 100 100 100 100 100 100 100 Intake 100 100 100 100 100 100 100 Test Solutions Reproduction (#young/female/7 days)

(%Effluent)

Data (replicate number) 1 2

3 4 [ 5 [ 6 7

8 9

10 Mean Control 30 34 34 31 32 29 32 31 29 31 31.3 Intake 33 29 33 33 33 36 33 32 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 = 1 00/LC50: TUc = 100/IC25 Page 7 of 99

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 August 14 - 21, 2012 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%

100 100 100 100 98 98 98 100.0%

100 100 100 100 100 100 100 Intake 100 100 100 100 100 100 100 Mean Dry Weight (mg)

Test Solutions(rpiaenm

)

(% Effuent)replicate number)

(%Effluent) 1

[

2 3

] 4 Mean Control 0.510 0.488 0.587 0.553 0.535 10.8%

0.673 0.559 0.631 0.594 0.614 21.6%

0.599 0.524 0.547 0.582 0.563 43.2%

0.592 0.516 0.548 0.588 0.561 86.4%

0.509 0.527 0.551 0.448 0.509 100.0%

0.550 0.565 0.526 0.494 0.534 Intake 0.62.1 0.573 0.647 0.482 0.581 IC 25 Value: > 100%

Calculated TU Estimates: < 1.0 TUc*

95% Confidence Limits:

Upper Limit: NA Lower Limit: NA

TUa = 100/LC5 0: TUc = 100/IC25 REFERENCE TOXICANT TEST RESULTS (see Appendix A and D)

Species Date Time 1 Duration I Toxicant I Results (IC25) I Pimephales promelas August 14 - 21, 2012 1225 Ceriodaphnia dubia August 07 - 14, 2012 1045 KC1 0.81 g/L NaCl 1.08 g/L Page 8 of 99

(0 (D

(0 0

1%

to PHYSICAL/CHEMICAL

SUMMARY

Water Chemistry Mean Values and Ranges for Pitnephales promelas and Ceriodaphnia dubia Tests, Non-treated Sequoyah Nuclear Plant (SQN) Outfall 101 performed August 14-21, 2012.

Test Sample ID Temperature (C)

Dissolved Oxygen (mgfL)

OH (S.U.)

Conductance Alkalinity Hardness Total Residual Initial Final Initial Final Initial Final (mtmhos/cm)

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

Chlorine (mglL)

Control 24.8 24.7 7.7 7.9 7.55 7.50 311 62 89 24.7 -

24.9 24.6 -

25.1 7.6 -

7.9 7.8 8.0 7.43 -

7.67 7.36 - 7.59 302 -

323 62 62 88 92 10.8%

24.8 24.6 7.9 7.8 7.52 7.46 291 24.7 -

25.0 24.3 -

25.0 7.7 8.0 7.6 8.0 7.44 -

7.62 7.33 -

7.55 285 298 21.6%

24.8 24.6 7.9 7.8 7.54 7.44 278 24.7 - 25.0 24.4 - 24.9 7.7 -

8.0 7.6 -

8.0 7.44 -

7.62 7.33 -

7.55 272 -

283

4.

24.8 24.6 7.9 7.8 7.53 7.44 252 432 13 24.7 -

25.0 24.4 -

24.8 7.7 8.0 7.6 8.0 7.45 -

7.61 7.34 -

7.52 246 -

259 24.8 24.6 7.9 7.8 7.51 7.42 198 S

86.4%

24.7 - 25.0 24.3 - 24.8 7.6 -

8.0 7.6 -

7.9 7.45 - 7.60 7.31 -

7.52 192 -

205 100.0%

24.9 24.6 7.9 7.9 7.51 7.46 182 71 72

< 0. 10 24.8 - 25.0 24.4 - 24.8 7.7 -

8.1 7.7 -

8.1 7.42 - 7.60 7.32 - 7.57 177 -

186 70 72 71 74

<0.10-

<0.10 Intake 24.9 24.6 8.0 8.0 7.51 7.45 182 69 73

< 0.10 24.7 - 25.0 24.4 - 24.7 7.7 -

8.2 7.9 -

8.0 7.43 -

7.61 7.33 - 7.54 176 -

186 68 70 72 74

<0.10-

<0.10 Control 24.8 25.1 7.7 8.0 7.55 7.51 311 62 89 24.7 - 24.9 25.0 -

25.2 7.6 -

7.9 7.8 -

8.2 7.43 -

7.67 7.28 -

7.59 302 -

323 62 62 88 92 10.8%

24.8 25.0 7.9 8.0 7.52 7.51 291 24.6 - 25.0 24.8 -

25.2 7.7 -

8.0 7.8 -

8.2 7.44 -

7.62 7.29 -

7.59 285 -

298 2!1 24.8 25.0 7.9 8.0 7.54 7.52 278 24.7 - 25.0 24.8 -

25.1 7.7 -

8.0 7.8 8.2 7.44 -

7.62 7.30 -

7.60 272 -

283 4.%

24.9 24.9 7.9 8.0 7.53 7.52 252 x

43.2%

24.7 - 25.0 24.8 -

25.3 7.7 -

8.0 7.9 -

8.2 7.45 -

7.61 7.30 -

7.61 246 -

259 8

24.9 25.0 7.9 8.0 7.51 7.51 198 86.4%

24.7 -

25.1 24.8 -

25.2 7.6 8.0 7.9 8.2 7.45 -

7.60 7.28 -

7.61 192 205 100.0%

25.0 24.9 7.9 8.0 7.51 7.52 182 71 72

< 0.10 24.8 -

25.1 24.8 -

25.2 7.7 8.1 7.9 8.3 7.42 - 7.60 7.29 -

7.63 177 -

186 70 72 71 74

<0.10-

<0.10 Intake 25.0 24.9 8.0 8.1 7.51 7.52 182 69 73

< 0.10 24.8 -

25.1 24.7 -

25.1 7.7 8.2 8.0 -

8.2 7.43 -

7.61 7.30 -

7.64 176 -

186 68 70 72 74

<0.10-

<0.10 Overall temperature (C)

Pimephales promelas Ceriodaphnia dubia Average 24.7 24.9 Minimum 24.3 24.6 Maximum 25.1 25.3

0,,

O

-I' (0

PHYSICAIJCHEM ICAL SUM MARY Water Chemistry Mean Values and Ranges for Pitnephalespromelas Test, UV-treated Sequoyah Nuclear Plant (SQN) Outfall 101 performed August 14-21, 2012.

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 (ng/L)

Control 24.8 24.7 7.9 7.8 7.59 7.49 304 61 84 24.8 -

24.9 24.5 - 24.9 7.8 -

8.1 7.6 -

7.9 7.50 -

7.65 7.34 -

7.61 298 -

316 61 62 84 84 10.8%

24.9 24.7 7.9 7.8 7.60 7.48 291 24.8 -

25.0 24.5 - 25.0 7.9 -

8.0 7.6 -

7.8 7.50 -

7.66 7.34 -

7.57 282 -

298 24.9 24.7 7.9 7.8 7.59 7.48 279 21.6%

21.6%

24.8 -

25.1 24.6 - 24.8 7.9 -

8.0 7.7 7.9 7.50 - 7.66 7.34 -

7.58 271 284 C,

24.9 24.7 7.9 7.8 7.58 7.47 254 43.2%

&1____

24.8 -

25.1 24.6 -

24.8 7.9 8.0 7.7 7.9 7.49 -

7.66 7.33

- 7.58 246 258 86.4%

25.0 24.7 8.0 7.8 7.56 7.46 203 24.9 -

25.1 24.3 -

24.8 7.9 -

8.1 7.5 7.9 7.48 -

7.63 7.33 -

7.57 197 -

207 100.0%

25.1 24.6 8.0 7.8 7.55 7.45 184 70 72

<0.10 25.0 -

25.1 24.2 -

24.7 7.8 -

8.3 7.5 7.9 7.47 -

7.62 7.31 -

7.55 178 -

188 70 70 72 72

<0.10 -

<0.10 Intake 25.0 24.6 8.1 7.9 7.54 7.46 184 70 72

< 0. 10 24.8 -

25.1 24.5 -

24.9 7.9 -

8.2 7.7 8.0 7.46 - 7.61 7.32 -

7.58 178 -

191 70 70 71 72

<0.10 -

<0.10

Note: Total residual chlorine was performed on non-treated Outfall 101 and Intake samples.

Overall temperature (C)

Average Minimum Maximum Pimephales promelas 24.8 24.2 25.1

SUMMARY

/CONCLUSIONS Outfall 101 samples collected August 12 - 17, 2012, 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 99

Appendix A ADDITIONAL TOXICITY TEST INFORMATION

SUMMARY

OF METHODS

1. Pimephales promelas Tests were conducted according to EPA-82 I -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 l-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. Pinephales 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 99

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 2510 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-821-R-02-013. Any known deviations were noted during the study and are reported herein.

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

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

2.

Standard Toxicant: Potassium Chloride (KCI crystalline) for Pimnephalespromelas.

Sodium Chloride (NaCl 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 99

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 l-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 99

Sequoyah Nuclear Plant Biomonitoring August 14 - 21, 2012 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 99

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 - August 17, 2012 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-H-130M Hypochlorite mg/L mg/L mg/L mg/L PF mg/L mgIL 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 1

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 I

I 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 1

0.016 0.067 0.023 0.020 Page 16 of 99

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 - August 17, 2012 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 01/31/2000 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 02/06/2000

< 0.002 0.027 0.009 I

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 1

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 I

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 1

Page 17 of 99

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 - August 17, 2012 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 01/02/2002 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 01/07/20021

<'0.0089 0.024 0.014 0.006 1

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 1

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 I

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 1

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

<0.01 18 0.026 0.020 01/15/2003

<0.0063 0.026 0.020 0.009 01/16/2003

<0.0034 0.026 0.020 01/17/20031

<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 Page 18 of 99

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 - August 17, 2012 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 1

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 10/10/2003 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 99

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 - August 17, 2012 Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-PF H-130M Nalco t-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 11109/2004

<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

-1 0.04 3 111/12/2004 1 <0.0253 0.048 1

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 I

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 I

Page 20 of 99

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 - August 17, 2012 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 mgiL 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 99

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 - August 17, 2012 Date Sodium Towerbrom PCL-PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSW Hypochlorite mg/L 222 mg/L mg/L A-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 05/12/10 0.0171 0.039 05/13/10 0.0041 0.039 05/14/10 0.0099 0.039 Page 22 of 99

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 - August 17, 2012 Date Sodium Towerbrom PCL-PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSW Floguard Hypo-mg/L 222 mg/L mg/L

-PF mg/L mg/L 73551 CT1300 mg/L 101 MS6236 chlorite TRC mg/L Copoly-DMAD Azole Quat mgiL mg/L Quat mg/P,

mgP mg/L Phos-meC EO/PO Quat Phos.- Phosphate TRC phate phate 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 0-0.04 05/05/2011 0.0179 0.04 05/06/2011 0.0089 11/06/2011 0.0168 11/07/20.11 0.0225 11/08/2011 0.0141 11/09/2011 0.0239 11/10/2011 0.0242 11/11/2011 0.0231 05/06/2012 05/07/2012 05/08/2012 0.041 05/09/2012 0.0145 0.041 05/10/2012 0.0298 0.041 05/11/2012 0.0174 08/12/2012 0.029 08/13/2012 0.0256 0.028 0.037 0.029 08/14/2012 0.0209 0.037 0.029 08/15/2012 0.0279 0.028 0.029 08/16/2012 0.0076 0.029 08/17/2012 0.0446 0.032 Page 23 of 99

Sequoyah Nuclear Plant Biomonitoring August 14 - 21, 2012 Appendix C Chain of Custody Records and Toxicity Test Bench Sheets Page 24 of 99

BIOMONITORING CHAIN OF CUSTODY RECORD Page _1_

of 'I U

Rent: TVA lRkject Name: Sequoyah NP Toxicity I%. Number: N/A Facility Sampled: Sequoyah NP NPDES Number: TN0026450 Collected By: pw-k% 1 ej r Environmental Testing Solution, Inc.

351 Depot Street.

Asheville, NC 28801 Phone:

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

FedEx UPS Bus Client Other (specify):

General Comments:

roolý 5*fYo

'-a

-f,P/1

.4+. 0 CCo, o,

.Fax:

828-350-9368 Container

.0.

Field Identification Grab/Comp.

Collection Date/Time&

Flow Rain Event?

Sample Description V.

CVolume (MGD) as Appropriate Laboratory Use Collected(

R

....~-.

s n hT

. E Tg.N r

.. iP". ".

p rIf Y es A rrival T emp.

B y T im e A ppear-

_______tt~~MW Inches Number ance 8 " 12 "t1-3 J-2I qi**.

Ii*

SQN-101-TOX Comp Lj 2(2.5gal) 1'I/

A o2/1,oi 17'_.

SQN-INT-TOX Comp 1

.x-Ot63 1(2.5gal)

)2?ol;: -.! i i

0Q Sample Custody - Fill In From Top Down.

wCturr se' iwpr-t,*'c. SAmAkes. ae.cewebl, Relinquished By (Signature):

Date/Time Received By (Signature):

Date/Time,

WLC.Pt A

0I-I*o t'v

-///

.o4:.0S 0

E 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.

MIS 1.11

ýIfl w

Whole Efflue'

-)ample Receipt Log Page.Z

Sample temperature performed usingSample Receiving Thermometer. SN 6338 0

(0 Date Time Received Received

Sa mple' Project Sample Received Received by from Temp. ( 0C) number number Sample name and description State Comments 08-13-12 1540 J. Sumner Sonic Delivery 0.8,0.7 8207 120813.01 TVA/ SON Outfall 101 TN 08-13-12 1540 J. Sumner Sonic Deliver 0.9 8207 120813.02 TVA / SQN Intake TN SOP G4 - Exhibit G4.2, revision 01-03-12

BIOMONITORING CHAIN OF CUSTODY RECORD Page 1

of 1.

2ent: TVA Environmental Testing Solution, Inc.

Delivered By (Circle One):

Wlject Name? Sequoyah NP Toxicity 351 Depot Street.

FedEx UPS Bus Client RA). Number: N/A Asheville, NC Other (specify):

2880 General Comments:

Facility Sampled: Sequoyah NP 28801 Teneral Comments:

NPDES Number: TN0026450 Phone:

828-350-9364

,,ke a

00,Jo, Collected By: t, 0 fl"Ž*.

Fax:

828-350-9368 Container A

Field Identification Number &

Flow RainEent?

FildIdntfiaton/

Grab/Comp.

Collection Date/Time Rain Event?

Sample Description Volume (MGD)

.""aLabspratoo aU)se Collected (Mark as Appropriate)

Start!~~

EndYes If Yes, No Trace ETS Log Arrival Tempp.

By, Time*

Appear-

i*W

° t ?

?YrJ.Str E

Inches

Number, ance SQN-101-TOX Comp 0700A 6"

0.

I" 2(2.5gal)

Y/*

l

[*

0 sQN-LNT-TOX Comp Wt d)o tv o Uve" 1(2.5 gal)

__Sample Custody - Fill In From Top Down 4

uokmR s

-s J

r5 RAE1 i.*.

Relinquished By (Signature):

Date/Time Received By (Signature):

Date/Time AtU

ýt".eS.

-T A

,hr

/03-e Skl._c C'r 6oC 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.

CTlS Whole EfflueK _.,ample Receipt Log Page 12>

~Iiii (0

0 (0(0

Sample temperature performed using Sample Receiving Thermometer SN 6338 Date Time Received Received

Sample Project Sample Received Received by from Temp. (0C) number number Sample name and description State Comments 08-15-12 0945 K. Keenan Fed - Ex 1.3 8215 120815.01 CORR ERI - AAF McQuay International NC 08-15-12 0945 K. Keenan Fed - Ex 0.7 8216 120815.02 Statesville Analytical - Cleveland WWTP NC 08-15-12 0945 K. Keenan Fed - Ex 5.5 8217 120815.03 United Water - Enfield WWTP NC 08-15-12 0945 K. Keenan Fed - Ex 5.5 8218 120815.04 United Water-Scotland Neck WWTP NC 08-15-12 0945

. Keenan Fed - Ex 0.3 8219 120815.05 Craven County Wood Energy NC 08-15-12 0945 K. Keenan Fed - Ex 0.7 8220 120815.06 Duke Energy - McGuire NS - Outfall 002 NC 08-15-12 0945 K. Keenan Fed - Ex 0.6 8221 120815.07 Progress Energy - Shearon Harris E & E NC 08-15-12 0945 K. Keenan Fed - Ex 0.5 8222 120815.08 Carolina Beach WWTP NC 08-15-12 0945 K. Keenan UPS 2.0 8223 120815.09 Morehead City WWTP NC 08-15-12 1553 J. Sumner TVA Courier 1.9/1.9 8207 120815.10 WA - SQN - 101 TN 08-15-12 1553 J. Sumner TVA Courier 2.0 8207 120815.11 ]TVA - SQN - Intake ITN I

SOP G4 - Exhibit G4.2, revision 01-03-12

BIOMONITORING CHAIN OF CUSTODY RECORD Page _1-of 4lent: TVA Wject Name: Sequoyah NP Toxicity Q*.

Number: N/A cc Facility Sampled: Sequoyah NP NPDES Number: TN0026450 Environmental Testing Solution, Inc.

351 Depot Street.

Asheville, NC 28801 Phone:

828-350-9364 FedEx UPS Bus Client Delivered By (Circle One):

Other (specify):__________

General Comments:

Q t- 0,( &,a/

7-V Collected By: Iw.;'f r6 n3in.p Fax:

828-350-9368 Container 1

8.',O.

N um ber &

Flow Rain Event?..:....

Field Identification /

Grab/Comp.

Collection Date/Time VumeRain Event?.M.D.

Sample Description Volume (MGD)

(Mark as Appropriate)

LaboratoryUsc Collected*(Mark.as..ppr-p..iate.

Laborator

.Use tW-:-

Sta...End

,t"

I Stets

Yes, No T

ETS Log Arrival Temp.

By

Time, Appearý-

Start__

End_ _

Yes Inches No Trace "Number ance.

SQN-101-TOX Comp 0 70o 67 Oo vr 2(2.5gal)

/

1 SQN-INT-TOX Comp O,- ec 0 C /j*.E 1(2.5 gal) 2 Sample Custody-Fill In From Top Down 4 CuV-abj Sf$t.5 1wrtA-SAKPLt' ft*1C.VCb Ji Relinquished By (Signature):

Date/Time Received By (Signature):

Date/Time

.SAM-s4 46t S-Pv.

Date/Tim 15e,-

qr 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.

~0 0

CC (0

X ýrs 11 --

Whole Efflue( '.ýample Receipt Log Page 12S

Sample temperature performed uslngSample Receiving Thermometer. SN 6338 Date Time Received Received

Sample Project Sample Received Received by from Temp. C0C) number number 08-17-12 0950 K. Keenan Fed - Ex 0.6 8215 120817.01 CORR ERI - AAF McQuay International NC 08-17-12 0950 K. Keenan Fed - Ex 0.6 8216 120817.02 Statesville Analytical - Cleveland WWTP NC 08-17-12 0950 K. Keenan Fed - Ex 2.0 8217 120817.03 United Water - Enfield WWTP NC 08-17-12 0950 K. Keenan Fed - Ex 2.0 8218 120817.04 United Water - Scotland Neck WWTP NC 08-17-12 0950 K. Keenan Fed - Ex 0.7 8219 120817.05 Craven County Wood Energy NC 08-17-12 1130 J. Sumner Murphy Courier 0.8 8226 120817.06 Duke Energy - McGuire NS - Upper Composite NC 08-17-12 1130 J. Sumner Murphy Courier 0.8 8226 120817.07 Duke Energy - McGuire NS - Lower Composite NC 08-17-12 1338 J. Sumner TVA Courier 2.2/2.2 8207 120817.08 TVA - SQN - Outfall 101 TN 08-17-12 1338 J. Sumner TVA Courier 2.4 8207 120817.09 TVA - SON - Intake TN SOP G4 - Exhibit G4.2, revision 01-03-12

TS Page Iof 6 Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephales promelas Client: Tennessee Valley Authority Facility: Sequoyah Nuclear Plant NPDES #: TN0020168 Project #:

A iJ1 County: Rhea Outfall: 101 Dilution preparation 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

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

Test information:

Organism age:

21) &ts O.z Randomizing template:

JeLLOQ.

Date and times organisms Oa

rh Incubator number and were born between:

shelf location:

Organism source:

AYb)( O k P e-Artemia CHM number:

LIA 6S2 Drying information for weight determination:

Transfer bowl information:

pH = -1. to 0 S.U.

Date / Time in oven:

06o..

1I%

Temperature = "S.*,

0C Initial oven temperature:

lb, C Average transfer volume:

Date / Time out of oven:

0. MLOck Final oven temperature: J Total-drying time:

".t*4 Daily feeding and renewal information:

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 2

-L I% O. 14-r 0w5 VLOI 11.01 1701.01l.2 O&F161 5

W-1R. 11-M IS Oh s A,

I %

__1*__ _'

I 0.16-m 2

0l- ____'_.

_____$_5

__ _\\___

l t*01l 0

5.

o-i'l1 6

0g**121. '-

g,-Ift 11.09 I 7.0,f n. do, a6--i7'IL u-.4 -

Ito(,

Control information:

Acceptance -riteria Summary of test endpoints:

% Mortality:

07,

< 20%

7-day LCsO

> I W7.

Average weight per initial larvae:

n3.SU W

NOEC 1CCW7.

Average weight per surviving larvae:

0. Sq

> 0.25mg/larvae.

LOEC

> 1007.

ChV

>1/MO.

IC 25 I

>1___T...

(I age 31 of 99 SOP AT20 -Exhibit AT20.3, revision 07-01-12

Page 2 of 6 Species: Pimephales promelas Client: TVA / Sequoyah Nuclear Plant, Outfall 101, Non-treated Date:

0&. 1q.- I,."

Survival and Growth Data Day CONTROL 10.8%

21.6%

/A B

C D

E F

G H

I J

K L

0 10DI(C/

0

/

/bo 10 o'o lio o/0o'ro0

__01_

t(0 /0 tol/0

/0

.10 /

/0 to(0 0

2

/0 to 10 lQ10 Ito (0

to t

10 to 10 (o

SIQ0 10 10 10 to 10 10 10 10 10 'o

/0 4_______

10 1o NO0 1 to Ito(

10 10 )

t0 o

10 to 2

/0

/0 10 10

/0 /0

/0

/0 /0

'0

/'0 (0

6

/0

/0 t0 10 (0 10 I0 10 (0

/0

/6 ID 6

11) to Io 'i o 101 10o fo o

t0 to

'ro 10 7

1o t)

/0 t

0 1O

/lb C) 1 10 CO

/

A = Pan weight (mg)

Tray color code::

L.;!,%*ý

t Analyst

wIl.y AA 13.-s 14.z4 13.Z4 14.10 13.B, I3.6 13-38 14.zs lZ.'e IS.

14.14 Date:

OA.n-.t-z B = Pan + Larvae weight (mg)

Analyst:

3"Lt Lo*o

%J.73 7.o.16 1I6V tq.q5 Zo.SS Z.Sq 1q.17 z2*33 19Sq 11,93 "u.S-Date:

09-zl.17.

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

S M 14-1.

SAI 6

6A0

.09 svi

(".m. 6..&6 Analyst:

k

_III Weight per initial number of larvae (mg)

= C / Initial number of larvae Hand calculated.

Analyst:

eV 0%

No

'I i'~p 4.

Average weight per Percent reduction initial number of from control (%)

0.614 larvae (mg) p -

i -

0. ts

-&.&7.

0.ADULD

- *.*a7.

I 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:

ge 32 of 99 SOP AT20 - Exhibit AT20.3, revision 07-01-12

I II I

I I

J.

I VI p

I w

I JETS Page 3 of 6 Species: Pimephales promelas Client: TVA / Sequovah Nuclear Plant. Outfall 101. Non-treated Date:

9O&.

Iq-I*.--

Survival and Growth Data Day 43.2%

86.4%

100%

M N

0 P

Q R

S T

U V

W X

0 0

to I olo to l

to to to IO U~

(

~

1,C) to 1

O 10( to to 10 10 / /0

/(0 I0 (

0 I)10 /)

2 j0 to to 10 10 to 10 1)0 to 0 o 0

10 3

I) to I

10 00 00

/0

/0to 0

0 to 0/0 4

t r/ I0 10 o 1

/0 t01 10o/

0

/0(0 5

ID 1

10 10 /0 10 10 10 (0

(0 10 6

M0 10 10 1 O tj ib Ib JO (0 "0 10 7

/) lb

(

1 1(

)

b 10 0 101 /

0

'o 0 to A = Pan weight (nag)

Tray color code:: Li-;,,k

I Analyst

-l hi.*

13.(.O I4.7 14.31 ml,.4 1

13.* )i

, '3.* o

3. 10 13.3m i4so Date: _______

B = Pan + Larvae weight (ing)

Analyst:

3Iq.a.

tq.3 iq.3z ZoL&5 1S

.'I 2

q.q z.

t..,qz Il C

J' 703r160 I

W.zI Date:

oft.27.I-t C = Larvae weight (mg) = B - A Anls:Hand calculated.*t 5.Z7. S.I

.tl 'A1.14..

6.1%;

S.%S SAI*

6.1%0 S..S2 I.."S S.0'6,.'1%

Anal yst J+/-ý Weight per initial number of larvae (rag)

C /Initial number of larvae Hand calculated.4%1*

4N Analyst'

%Y l"

0 Average weight per Percent reduction initial number of from control (%)

o.

%L'.-

?.L1

0.

O.'606

-S1.

".7.

larvae (Mg)

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

Comments.

Sage 33of 99 SOP AT20 - Exhibit AT20.3, revision 07-01-12

ETS Page 4 of 6 Species: Pimephales promelas Client: TVA / Sequoyah Nuclear Plant, Outfall 101, Non-treated Date:

Day 100% Intake Y

Z AA BB 0

C0 Io to0 10 1

I0 (o to 10 2

1) to 10 10 3

(Q '/0 /0

/0

/D1 /0,/0 5

/o0

/0

/0 t0 6

(o 0 10 (0

A =Pan weight (ng)

Tray color code::

V Analyst:

M"

'14.o 14.11 1'4.24 Date:

Of.t2.

B = Pan + Larvae weight (mg)

Analyst:

M

.2.

Z0.47 T..7-L1.11 Date:

oS-z7..

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

S.S'" ',.i

..i h otj Analyst-Weight per Initial number of larvae (mg)

= C / Initial number of larvae Hand calculated.

Average weight per Percent reduction initial number of from control (%)

0 *-96 0A 7.

larvae (mg)

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

Comments:

ge 34 of 99 SOP AT20 - Exhibit AT2O.3, revision 07-01-12

,E Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated August 14-21, 2012 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 n umber.

8207 Not fnr Compliance Assessment. Interal Laboratoru OC Coeatratia(%)

Replickte nlitnambered Flnatoumberftla,-ae APa welght(mg)

BPaa+Larvae Larvae wetghtrt(m)

Wdght /Surviitng Mea iget I Coeffilaetntuarraia Weight/ti-lai mahber MMaa adval Mea weightInlal Cliteataf P-er1a eed.dm f.

larvae wseight (.0g)

-A-0 nberoftaa (rag)

SvMngtumbero" tMa' ea alarva(rg)

(*A) 0a--re-rhlauea arao (5u..es ooa rsI (%)

lourvartag) a~dktA (01g) aeao.irt A

10 10 14.47 20.30 5.83 0.583 0.583 a

10 10 13.58 19.73 6.15 0.615 0.615 Control C

10 10 14.24 20.16 5.92 0.592 0.584 5.1 0.592 100.0 0.584 5.1 Not applicable D

10 10 13.24 18.68 5.44 0.544 0.544 E

10 10 14.10 19.45 5.35 0.535 0.535 1"%

F 10 10 13.86 20.55 6.69 0.669 0.669 100.0 0.635 10.7

-8.8 G

10 10 13.68 20.54 6.86 0.686 0.686 H

to 10 13.38 19.87 6.49 0.649 0.649 1

10 10 14.25 20.33 6.08 0.608 0.608 J

10 10 12.95 18.54 5.59 0.559 0.559 K

10 10 15.67 21.93 626 0.626 0.626 L

10 10 14.97 21.83 6.86 0.686 0.686 M

10 10 13.50 19.32 5.82 0.582 0.582 43.2%

N o0 10 13.60 19.23 5.63 0.563 0.562 9.6 0-563 100.0 0.562 9.6 3.8 0

10 10 14.75 19.62 4.87 0.487 P

10 10 14.31 20.45 6.14 0.614 0.614 1

Q*

10 10 12.64 18.78 6.14 0.614 0.614 R

10 10 13.87 19.42 5.55 0.555 0.555 S

10 10 13.79 19.42 5.63 0.563 0.563 T

10 10

.14.52 21.42 6.90 0.690 0.690 U

10 10 13.50 18.82 5.32 0.532 0.532 V

10 10 13.70 20.43 6.73 0.6730.7 100%

W 10 10 13.37 18.43 5.06 0.506 0.571 12.9 0100.0 0.71 12.9 2.2 X

10 10 14.50 20.21 5.71 0.571 0.571 Y

10 8

13.84 19.21 5.37 0.671 0.537 100%Intake Z

10 10 14.78 20.47 5.69 0.569 0.620 11.2 0.56995.0 06 1

.7

-0.4 AA 10 10 14.71 20.22 5.51 0.551 BB 10 10 14.24 21.11 1

6.87 060.687 0.6 Outfall 101; Duanett's MSD value:

PMSD:

intake Dunnetas MSD.value:

PMSD:

0.0994 17.0 0.0726 12.4 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 (10th percentile) = 128/6.

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. 2001 b. Final Report: Interlaboratoty Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity To. Me hn4,_ Vc zt I

n. 2-Appemii. EPA-t2 t-8-01-004 and EPA-821-B-01-005. US Enviromnental Protection Agency. Cincinnati, OR File: sqn101_081412data.xlsx Entered by. J. Sumner Reviewed by: G

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated August 14-21, 2012 Statistical Analyses aETS 6

Larval Fish Growth and Survival Test-7 Day Growth Start Date:

8114/2012 Test ID:

PpFRCR Sample ID:

VA I SQN 101 End Date:

8/21/2012 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Dlscharge Monitoring Report Sample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species:

PP-Pimephales promelas Comments:

Non-treated Conc-%

1 2

3 4

D-Cbntrol 0.5830 0.6150 0.5920 0.5440 10.8 0.5350 0.6690 0.6860 0.8490 21.6 0.6080 0.5590 0.6260 0.6860 43.2 0.5820 0.5630 0.4870 0.6140 86.4 0.6140 0.5550 0.5630 0.6900 100 0.5320 0.6730 0.5060 0.5710 Intake 0.5370 0.5890 0.5510 0.6870 Transform: Untransformed 1-Tailed Isotonic Conc-%

Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD Mean N-Mean D-Control 0.5835 1.0000 0.5835 0.5440 0.6150 5.089 4

0.8127 1.0000 10.8 0.6348 1.0878 0.6348 0.5350 0.6860 10.744 4

-1.242 2.410 0.0994 0.6127 1.0000 21.8 0.6198 1.0621 0.6198 0.5590 0.6860 8.465 4

-0.879 2.410 0.0994 0.6127 1.0000 43.2 0.5615 0.9623 0.5615 0.4870 0.6140 9.607 4

0.533 2.410 0.0994 0.5835 0,9524 86.4 0.6055 1.0377 0.8055 0.5550 0.8900 10.256 4

-0.533 2.410 0.0994 0.5835 0.9524 100 0.5705 0.9777 0.5705 0.5080 0.6730 12.860 4

0.315 2.410 0.0994 0.5705 0.9312 Intake 0.5860 1.0043 0.5860 0.5370 0.6870 11.706 4

Auxiliary Tests Statistic Critical Skew Kurt Stlapiro-Witk's Test Indicates normal distribution (p > 0.01) 0.98195 0.884 0.03805 -0.4693 Bartletts Test Indicates equal variances (p = 0.82) 2.21405 15.0863 Hypothesis Test (1-tall, 0.05)

NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnetts Test 100

>100 1

0.09943 0.17041 0.00332 0.0034 0.4689 5,18 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point SD 95% CL(Exp)

Skew IC05 87.934 IC10

>100 IC15

>100 1.0 IC20

>100 0.9 lC25

>100 0.8 IC40

>100 IC50

>1000.7 0.6

, 0.4 0.2 0.1 0.0

-0.2 0

50 100 150 Dose %

Dose-Response Plot UZ.5 0.7 0.8 0.5, 0

80.4 0*.3.

0.2 0.11 1-tall, 0.05 level of significance 8

m i~4.

6 o

0 S_

(tage 36 of 99 File: sqn101_08l412dt a.xlsx Entered by. J. Eumner Reviewed by: -V

TVA / Sequoyah Nuclear Plant, Intake Non-treated August 14-21, 2012 ErrvtlronmentalTestlig Solutlonsý Inc.

Statistical Analyses Larval Fish Growth and Survival Test-7 Day Growth Start Date:

811412012 Test ID:

PpFRCR Sample ID:

TVA / SQN 101 - Intake End Date:

8/21/2012 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species:

PP-Pimephales promelas Comments:

Non-treated Conc-%

1 2

3 4

D-Control 0.5830 0.6150 0.5920 0.5440 10.8 0.5350 0.6690 0.6860 0.6490 21.6 0.6080 0.5590 0.6260 0.6860 43.2 0.5820 0.5630 0.4870 0.6140 86.4 0.6140 0.5550 0.5630 0.6900 100 0.5320 0.6730 0.5060 0.5710 Intake 0.5370 0.5690 0.5510 0.6870 Transform: Untransformed 1-Tailed Conc-%

Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD D-Control 0.5835 1.0000 0.5835 0.5440 0.6150 5.069 4

10.8 0.6348 1.0878 0.6348 0.5350 0.6860 10.744 4

21.6 0.6198 1.0621 0.6198 0.5590 0.6860 8.465 4

43.2 0.5615 0.9623 0.5615 0.4870 0.6140 9.607 4

86.4 0.6055 1.0377 0.6055 0.5550 0.6900 10.256 4

100 0.5705 0.9777 0.5705 0.5060 0.6730 12.860 4

Intake 0.5860 1.0043 0.5860 0.5370 0.6870 11.706 4

-0.067 1.943 0.0726 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.88547 0.749 1.36005 1.98359 F-Test indicates equal variances (p = 0.20) 5.37752 47.4683 Hypothesis Test (1-tail, 0.05)

MSDu MSDp MSB MSE F-Prob dl' HomoscedastictTest indicates no significant differences 0.07258 0.12439 1.2E-05 0.00279 0.94881 1,6 Treatments vs D-Control Dose-Response Plot 0.8 0.7-0.6 0.5 0

0.3 0.2 0.1 0O 1-tail, 0.05 level of significance 8

Nq N

(I age 37 of 99 File: sqn10I_081412dataxlsx Entered by: J. Sumner Reviewed by: I F

Page 5 of 6 Species: Pimephales promelas Client: TVA / Sequovah Nuclear Plant Outfall 101. Non-treated Daily Chemnsfr:

Date:

ot'4,V*.-

I r

I Day I

(Analyst identified for each d. performed PH D.O. and conducvit measurements only d

t'[

,D.O. and conductivity measurements onli 0

T Analyst MI 1

I 2

=--4 Concentration

[Parameter I nH(S.U.)

I DO (ma/L)

IConductivity CONTROL [

CONonTR

Alkalinity Non-treated (mg CaCO3/L)

Z Hardness em CaRCOZ.)'

Temperature (0C) pH (S.U.)

DO (meL/

10.8%

Conductivity (inmhos/cm) 1 0

q7 V14 VA.1%

_LS.0 7.C?-

1 "4!6 1 47 D~E I 7. Si II

Temperature (M)

S U

nH (S.U.)

J DO (mF_/)

-7.7 21.6%

Conductivity IS (gmnhos/cm)

_20 I

IS

Temnerature (*0)

I_*.

II tuq.

I

~

II I~

-I I

pH (S.U.)

-741' 1.z 1 i7.

So 751l t

II* -

DO (mg/L)

-'--I

.1. ~

43.2%

Ts Conductivity.

(gmhos

)

Temperature (0C)

"1,"1.1 I

...,.1tLI U

U __________

a PH (S.U.)

7.31l 86.4%

Conductivity z6

&+/-mhos c)

I6 9.01..

255 7-55 1.51 7.51 1

.9.0

%-b.

Temperature (00)

"u4.'! H "LL.I.3h

-L-

-1

'V+I DH (S.U-

"7.C5 I1 -7.'IZ I.

4 Z.

DO (mFJL) 100%

Conductivity (pinhos/c)

Alkalinity

Hardness (mng CaCO3IL) i

TR chlorine (mg/L) 0O. 1 E 9.'

_10 o.I'D T 0ISI

Temperature (0C)

PH (S.U.)

DO (ma/)2 Conductivity (iimhos/cm) 100%

Alkalinity Intake (Ing 03L)

Hardness 1

(Ing Chlorine (m/L)

TR chlorine (mg/L)

0. to II I

r*

7-9Z I

-- Ik-----

I St

'0.104

".Il t.

I I

Te'm".

U I

" I° 5 II M

S U

I Initial Final Initial Final Initial Final

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1.

Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet SOP AT2O - Exhibit AT2O.3, revision 07.-01-12

ý7jage38 of 99

.j~.

ETS Spec Cliei Page 6 of 6 ies: Pimephales promelas nt: TVA / Secuo ah Nuclear Plant. Outfall 101. Non-treated Date:

c* t."i'A l

Dae 1

.1 I

I I

I Day (Analyst identified for each day. performed pHi, D.O. and conductivity measurements onlv.,

7 iB Analyst F-4 I6 Concen-Parameter traon__

PH (S.U.)

DO (mg/L)

I 7.Co 5

I II q.*l'l II II

~1.~

I 1..'

II CONTROL Non-treated Conductivity 0

(iimhos/crn) 80

Alkalinity (Mg CaCO3/L)60

Hardness

(-g CaCO3/L)

31S

-7 St "310 37.3 II I

Temperature (0C)

%A I.

I.

71 IP I

-L*'

I

-,/Ia.

I

~**

41.

I

~*-.

~

pH (S.U.)

/;iJS*'l" 7.41

-7. 45 7-51 "7..S

1. QS 7.3 3 10.8%

DO (mg/L)

'6'0____

Conductivity 2`65a..

us Z%5*7t

Temperature (*C)

'MA

.1. t IM- !6 14.1)

"I, k

'1%. 2 i

i p

PH au.)

T

-7.

-7.57.

1.33 1

.,7 SL-7.i DO nutagv 7.9

-7 21.6%

Conductivity z st pH (S.U.)

i' "7.£1.

7.~l

-7.51

-7.SL I ".C;

'.z ":4 i DO (mnA/L' 43.2%

Conductivity (gmhos/cm V4m 1 -7.4 7.7.5.

Temperature 4-nH (S.U.)

7-4.3 DO (mF/L) 86.4%

Conductivity O~mhos/cm) 0-O

~

11.c I77 6"

"14 1.94.

-7C

.aQQ

'7_441

-797

.M

-7.S I

TemperatuJre C*C) 6 S

i U -

DR (S.U.)

DO (mR/L) 100%

Conductivity

Alkalinity (mg CaCO3IL)

Hardness (Mg CaCO3/L)

TR chlorine (mg/L)

Temperature (*C) i 21 l7'~

'Vt-

-wI

~O. /0 II I

2*.D

-U 100%

Intake Conductivity

Alkalinity i

Hardness

(*ag CaCO3/L)

TR chlorine (mg/&)

"IW

...a.1

-*-a.

V.I.I I*4t.-.

I

.I Iitl Final I

Initial F15I I nritial1 Final InI6tial IF Finial

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1.

Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet SOP AT20 - Exhibit AT20.3, revision 07-01-12 i

age 39 of 99

(D*

0O SEiTS b

EnvironmentalTestig Solution,' Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated August 14-21, 2012 Pimephalespromelas Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Daily Chemical Analyses Project number:

8207 Day 6 Concentration IParameter I

.~

I I

pH (SU)

Control DO (mg/L)

Conductivity (imhos/cm Alkalinity (mg/L CaCO3)

Hardness (mg/L CaCO3)

Initial F Inal Initial I

Final Initial I

Final I nit 7.49 7.49 7.60 745 7.67 75 7.6F 7.8 7.6 78 7.6 7.

302

,315 310 WE 62 88W fl I

Vi...l I

A h enpraur(~)74 246 29 4724 27 28 27 4

4 27 21 247 2.

H(SU) 762 744 744 752 745 752 757 432 5

4 5

5

.5 73

______Temperature (C) 24.7 2446 2509 24.7 24.81 2467 2498 2497 24.8 24.6 24.7 2024.8 24.31 PH(SU) 7.62 7.44 7.44 7472 7.5 7.50 7.59 7.43 7.52 7335 7.5215 7.55 7.33 DO (mX/L) 7.7 79 8.0 7.8 7.9 789 790 7.6 7.9 768 80 787.8 8.0 210.6%

Conductivity (jLmhos/cm) 297__

294___

294__

285__

285_298

______Temperature (CC) 24.7 24.4 25.0 2467 24.8 24.6 24.9 24.9 24.8 2456 24.7 2480 24.9 24.4 pH (SU) 7.61 7.42 7.45 7507 7.51 7.50 7.59 7.42, 7.51, 7.37 7.51 7.52 7.56 7.34 4.% DO (mxnL) 7.7 78.0 9

8.0 7.8 7.9 7.6 7.9 7.6 8.0 7.8 7.8 8.0 2.% Conductivity (gmhos/cm) 280__

283__280 279 278 276 282 Temperature CC) 24.7 24.4 25.0 24.6 24.8 24.8 24.9 24.6 24.8 24.5 24.71 24.8 24.9 24.4 pH (SU) 7.60 7392 7.45 7.51 7.51 7.48 7.58 7.42 7.451 7.34 7451 7.52 7.53 7.,314 86O4% DO(

L) 767 7.8 8.0 7.9 8.0 7.8 8.0 7.6 7.8 7.7 8.0 7.8 7.8 7.9 4.% Conductivity (IRmhos/cni) 254_255 195 2492 24 241~6 2I5~

_______Temperature CC) 24.7 24.43 25.0 24.6 24.8 24.6 24.9 24.6 24.8 24.7 24.7 2468 24.9 24.5 piH (SUY) 7.60 7.42 7.42 7.54 7.53 7.49 7.57 7.49 7.44 7.38 7.44 7.57 7.54 7.32 DO (mg[L) 7.7 7.8 8.0 8.1 8.1 7.9 8.0 7.7 7.8 7.7 8.0

78.

7.8 7.9 8.% Conductivity (,Imhos/cm) 183 183 186 181 1796 177 183

_______Temperature (OC) 24.8 24.4 25.01 24.7 25.0 24.6 25.0 24.8,

2.

24.

7 24.

8 24.7 24.9 24.5 HIE (SU) 7.61.

7.42 7.44

-7.50 7.52 7.50 7.58 7.4 7.43 7.39 7.434.4 75 7.33.

DO (mg/L) 7.7 79 80 8.0 8.2 8.0 8.10.

7.9 7.9 8.0 8.7.

.0 C'onductivity (jnhos/cm) 183 186 185 kJd 182 181 17678 100% Itk Alkalinity (mgtL CaCO3) 6826 70 7

Hardness (mg/L CaCO3) 72 74 72 Total Residual Chlorine (niL

<0.10

.<0.10

<0.10 ITemperature CC)

2.

24.

4 24.9 24.7 25.0 24.6 25.0

2.

25.0 24.7847

2.

24.9 24.7 File: sqn101_081412chem.xls Entered by: J. Sumnner Reviewed by: A

Page 1 of 7 OETS Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1002.0)

Species: Ceriodaphnia dubia Client: Tennessee Valley Authority Facility: Sequoyah Nuclear Plant NPDES #: TN0026450 Project #:

&,Ldl County: Hamilton Outfall: 101 Dilution preparation information:

Comments:

Dilution prep (%)

10.8 21.6 43.2 86.4 I100 Eftluent volume (mL) 270 540 1080 2160 2500 Diluent volume (mL) 2230 1960 1420 3406 0

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

Test information:

Oranism age:

< 24-hours old Randomizing template color.

6'0 $.b Date and times organisms were born ol't

" i2..O-04o0 lb C Incubator number and shelf between:

location:

7-6a-Culture board:

a2 It

r.

locaton Replicate mnuber 1

2 3

4 1S-6 17 8

9 10 Culture board cup number [.

I

-W I U M Mt YWT1batch:

1.U Transfer vessel information:

PH = -611 S.U.

Temperature = L C

Seleasm batch:

Average transfer volume (mL):

05

.S 6.

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 ot.N-r IN?

O -I t.rLo041

%2 tOO it.Q_.

1 ~ ~

~

1S 2ri~r I2.oels.0%

Xtt~

2 o1.n..0' O40 oIq'.

1T-tjOtt. IU t lo t. I It 4

Os.lo. I"L 17_.oeI1.QJ I"

IS Itt S 1 doI1.11.

iOS"_

1_9 11-__I,-

'L_

a %

091t-l.ot L.

60 1 6

S-20.r01 A~l IL. I 1'P l1% Rs 1--61.)I ni I.

7 Of" Lt-1-2 h

I Control information:

Summary of test endpoinils:

Control-1 Control-2 Acceptance criteria

% of Male Adults:

07-07,

< 20%

7-day LC5s

>00Q1.

% Adults having 3'( Broods:

1001

fool,

Ž80%

NOEC 1001,,

% Mortality:

17.

07.

< 20%

LOEC

>1861.

Mean Offspring/Female:

31.7.

1

,31.3 15.0 offspring/female ChV

> 10.

,% CV:

(,'A"27.

I S.6#7.

< 40.0 %

IIC25

') 100 1J.

(

age 41 of 99 SOP ATI I - Exhibit AT 11.2, revision 07-01-12

ETS Species: Ceriodaphnlia dubta Client: TVA / Seauovah Nuclear Plant, Outfall 101 CONTROL-1 Surv.

Page 2 of 7 Date: __

-I_

ival and Reproduction Data Replicate number Day 1

2 3

4 5

6 7

8 9 110 1

Young produced 0

C)

C D 0

0 1

Adult mortality

\\-

k..

I 2

Young produced l

0 1

1 1

I-Adult mortality J

... [

- le

-I-3 Young produced 0

I C

Adultmortality I

_ -1 I

U I

- f 4

Young produced C.,

1

,[

O__

Adult mortality

[

U

5 Young produced

%0i 1 I

Final Adult Mortali ty t..

I Xfor3 u

Broods 1

0-0 0

0 0

O I

Note: MeAdult morta litve D = dead), SB = epit brood (single brood

%plit between two days), CO cariy oere (offcpnng earied over with adult durig trasfer).

tenOfping/emale:

_',L CONg: 10.8%

Survival and Reproduction Data 1 ______

Replicate number________

Day produce 2

3 4

6 7

8 9

0 1JYoung prd__

__[*

O Adult mortality

21[Young produced 0

0 0I QI j

CL 10 C

JAdult mortality VwV *IL.tL

_UJ.

j.._

... 1L

U

]

No te:ng rdueAdult mortalit(L=

fiv.

D"- ead B spli bro snl ro pltbtentody), CO - carr over (f(n care vrwt dl uigtase) 4 Youn produced O..c.

0 0

0 J Adult mortality M_

.rai I

Young produced 0N 3

0 C

Adult mortality jZ[__

1.

I.

C 2

Young produced C)6 0t 1 a.i IAdult mortality I

-C]~

Lj\\-

~C U

I 6

Young produced n

0) 0

.0

.c.10 m

F i

__fAdult nortality.I 2

L-I~P1_~~~H L-tL4k 4

Young produced

-6 tre T

It4 I

Adult___

mo talityt..

t Totalyoung produced 3 S J S I

IL Final Adult Mortality

.U -

T U..

Finot Adult Mortality

\\

C -

-eCo L.-

Qr o

Note: Mdull mortality (L - live. D - dead), SB -eplit brood (single brood split betwoeen two days), CO -cam over (offispijer caruied over with adult during rasfr Concentration:

i I

age 42 of 99

% Mortality:

Mean Offspring/Female:

S'1

% Reduction from Control-1:

-. O7 SOP ATI I - Exhibit ATI 1.2, revision 07-01-12

0ETS Page 3 of 7 Species: Ceriodaphnia dubia Client: TVA / Sequovah Nuclear Plant. Outfall 101 CONC: 21.6%

Suri' Date: ______I__"__L_

ival and Reproduction Data Replicate number Day 1

2 3

4 5!

6 7

8 9

10 1 Young produced 0

C_ 1f 0 0 1 0) 0 _

2 Young produced 0

0 0____

JAdult mortality U

.L.__

3J*Young produced W

0 0

0 Adult mortality I

I L-I'.--1 4

Young produced

.S S

S.

w a

I I

SAdult mortality RE Yonpone I *'- I I I___

I t'- I_'l

[

5 Youngproduced I I.

[

Adult mortality

-I'

'I UI' 6

Youug produced 1

1C) 0 0

C)

I cc >

7_..

Adult mortality I U

7]

Young produced 40 1-15

~~'

Total young produced I

3 I

I ?

Final Adult Mortalit Note: Adult mortality (L li D -dead), SB -split brood (sge brood split between two days), CO ca over (offsorimit carried over with adult durinatranafer).

Concentration:

% Mortality:

07.

Mean Offspring/Female:

,32, 1

% Reduction from Control-1:

l7.

CONw: 43.2%

Survival and Reproduction Data Replicate number Day 1 1 2

3 4

5 6

7 8

9 10 1

edYoung produce 0

)

(

].Adult mortality [12.

7&7 1.

U.

L.. I L..

=.

LL...

Adult mortal'yit 2

Young produced I C

Q 1 5 1

0 1

1 4

on rdcdAdult mortality I

L L.

I_ --.

I t._

t I

I..

2 Young produced I1 l

()

I C I Cz Adult mortality L

L 4

Young produced SC) kA 10 S1 6...oJ~

1 So.I 4C S~..

__[Adult mortality C_ C...I

~

I 4

Yung produced

%I L

Adult mortality L

L __

UU.

6

Young produced 1

)

C)

I Adult mortality L-1 A-f --/

7]

Young produced 1

l 1

Total young produced

-.b Iq -&- 1311 31 a% 1 :31 13'4 13" 1 as k_

L..

L_

K_ I C

I_

L_

Notef

.4

_/

UIUSI.ffWUPt JUIUY.

SUU)J.

-A - an k

flIý i V~

O Ot OU.'. H I Concentration:

-1

% Mortality:

Mean Offspring/Female:

3.

% Reduction from Control-I:

7.

SOP ATI I - Exhibit ATI 1.2, revision 07-01-12 (Iage 43 of 99

A!i-0 S*ETS a0 Page 4 of 7 Species: Ceriodaphnia dubia Client: TVA / Seauoyah Nuclear Plant Outfall 101 CONC: 86.4%

Surv, Date:

0'1 1+_17._

ival and Reproduction Data 1

Replicate number Day 1

2 3

4 T 6

17 18 9

10 I

Young produced C

)0 C

)

C

_J Adult mortality Fk 4_

L, 0

LJ L*

L_

[

1 Youngproduced A1 r..

0,"I 0 Q I 0 Q

Adult modaity

\\._I \\_,

=T T.

I °

=

3 EYoung produced 1 01 D.Q I c>.

o

.....Q.

Adult mortalityI,

J L.. I_

4 1 Young produced I

.utmortality

_L 7 Young produced i,_,,\\I 1"

'J I'-1

'L Adult mortality I--I N" I

I 1

6 Young produced C)

I Adult mortality i

[I' I° I1

-I a I-7 Youngproduced IS

%4% 14 lt0 Total young produced "A

S4 6

""S X1 Final Adult Mortality

--1

\\-

\\_

C I

Note: Adult mortality (L - live, D-dead), SB -split brood (single brood split between two days), CO - can over (offspri carried over with adult dourim transfer.

Concentration:

% Mortality:

7.

Mean Offspring/Female:

% Reduction from Control-1:

-I.S7.

CONC: 100%

Survival and Reproduction Data Replicate number '_"

Day 1

2 3

4 5

6 7

8 9

10 Young produced1 Adult mortality 1-2

_ I' L_ I.'_'

L I L_

L-IL--

2 Young produced C) c))

Adult mortality L

_[0 [

10 I I

o 3

AYunglproduiedt 1 L3 C) 1 0 1 [

1 0 1

- 1 Adult mortality I_

I-I L_

I L_

I*-

4.

Young produced Adult mortaliyI I V[ '1 1 '-

J

_l

__I 5

IYoungproducedI-I I

VWI Cm J1 ('

Adult mortality I 1_ x-I x---I I

l I_ -ILI 6

Young produced

[

l o0 0_0__

_i 0.

Adult mortality I_

L k

L-I-----

7]

Young produced iI 4e I

1 i 1 9, 19 1 -1 "7

.O it 19 Total young produced U

T 31 3-1 3

. 3

,8 36 YI 31 Z 8 Final Adult Mortality "C.._ -c.

N.

- L_

-L_

C.

C AW*s: Adult mortality (L i-live, D - dead), Sn - split brood (single brood split between two days). CO = car over (offssrint carred over with adult durint transer).

i Concentration:

i

% Mortality:

Mean Offspring/Female:

% Reduction from Control-1:

1.0Z SOP ATI 1 - Exhibit ATI 1.2, revision 07-01-12

age 44 of 99

ETS 6

U~m*TstfDfm t sawl=*M Page 5 of 7 Species: Cerlodaphnia dubia Client: TVA / Seauovah Nuclear Plant. Outfall 101 CONTROL-2 Surv Date:

O.t'it.

ival and Reproduction Data Replicate number Day-1 2

3 4

5 6

7 8

9 110 I Young produced 2

-Yugpodcd 0

0___

C)

C 2

Young prdue 1010_oi,..C.[o..

0.1..

Adult mortality

\\L L..J-

.\\J_

j.__

3 ~F r_

L 0

4 Young produced d*t 1=

Adult mortality L_

- L I i-I

-. I 5

Young produced

IC)

I___

10 10 I'*3 A dult mortality I~ %.

L 1

_j L_~

%[..

6

[Young produced 0

T01 0

1 Adult mortality J__

7 Youngproduced Total young produced A".".

Final Adult Mortality JC:

I I

X for 3Y Broods

y.

Y.

5

.2-.

5Z 5Z Note: Adult mortality (L - live, D dead). SB = split brood (single brood split between two days), CO - carry over (offspring carried over with adult durin transfer).

I Ccent.ration:

_Mortal__:

_I IMean Offspring/Female:

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

2 3

4 5

6 7

8 9

10 1

Young produced 1Q5 1 C)

I _

(.2 0

0 L I Adultmortality L.

2 Young produced 0

0 Adult mortality

).. -

I

'[-

1 Lc L.

3 Young produced mrai O

p 1

_OI O

cI.

4 IAdult mortality

-,Ij_,,..

L I kI I'.

L_

4 Young produced L

I I1 SI S1i[S ISI I I A dolt mortality Q

L IL 6

Young produced [

1 T! -W I..

I

A__

dolt mortality [

[~I~*I-

-I'_~i'

_=IL.

61 Young produced [C)j1c01TOK1 0

1 ZII0iC A__

dult mortality

.~L L

7 Young produced [6 IS 10 1

.~

~

-1 Total young produced 31 3

ac3 3-6 "3,

32. 1 31-

'Aq Final Adult Mortality

\\_

L LZ

=

-C7

ý Nvote: Adult mortality (L -= lte. D dud), Sa - spit brood (tingle blood split between two days),

) -c an over t(oftain carred over with adult duttna transfer).

Concentration:

% Mortality:

07.

Mean Offspring/Female:

% Reduction from Control-2:

I SOP AT1I1 - Exhibit AT 11.2, revision 07-01-12 I

age 45 of 99

(ia!

(0 (0

0:T a

E&trn*"nsrtTeUng SIWUmInc,. c TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated August 14-21, 2012 Verification of Ceriodaphnia Reproduction Totals Control-1 Day Re plicate number Total D

1 2 03 4 0 6 0 7

0 9

10 2

0 0

0 2

0 0

0 3

0 0

0 4

5 4

4 5

5 5

5 6

4 6

49 5

111 11 10 "12 10 9

12 11 11 11 108 L6 0

0 0

0 0 0

0 0 M 0

0 7

14 18 15 16 16 14 17 15 15 15 155 Total 30 33 233 3

31 28 34 32 30 32 312 10.8%

DyRe plicate number Total 1

2 3

4 5

6 0

7 0 0

9 10 1

0 0

0 2

0 0

0 3

0 0

0 4

L5 6

4 4

3 6

6 4 L6 5

5 3

51 1

13 12 11 12 12 10 1

12 12 12 1 19 3

0 0

0 0'

0 1

0 00 0 0 0

o0o0o 0

0 7

18 16 18 17 17 19 17 16 14 15

,167 Total 36 34 33 31 30 37 34 33 32 30 337 21.6%

Day

~Re plicate numberToa Dy 1 2

3 4

5 6 17 18 19 110 Toa 1

0 0

0 2

0

.0-0 0

0 0

0 50 0

0 0

3 0

0 0

4 5

5 5

5 6

5 5

4 4

4 48 5

13

!11 12 10 10 13 13 13 11 13 119 6

10 10 0

0 0

7 118 1 15 F6 16 14 19 17 1-6 1-7 1

6 Total 136 131 33 131 30 137 _L5

_L3 32 130 1328 43.2%

DyRe plicate number Total S 1 0

0 0

0-0 0

0 0

0 10 0

2 0

0 0

3 0

0-0 0

0 0

0 4"

1 2

3 4

5 1 6 7 1 85 1 1 5 0 5

13 12 12 12 11 11 12 14 12 12 121 6

0 0

0 7

18 17 17 15 15 15 19 15 18 18 6

Tot-s-,

M 1 541 M i M13131 371=

413r]

31133s 86.4%

Replicate number Total Day 1

2 3

4

_5 6

7 8

9 10 1

0 0-0 0

0 0

0 2

0 0

0 3

0 0

.0 0

0 0

0 4

5 7

5 5

4 5

6 5

6 6

54 S

13 12 12 11 12 12 14 1L2 13 13 124 6

0 0

0 7

16 15 19 18 19 16 14 18 17 18 170 Total 34 34 36 34 35 33 34 35 36 37 348 100%

Replicate number Total Day 1

2 3

4 5

6 7

8 9

10 T

0 0

0 2

0 0

0 2

0 0

0 3

0 0

0 7

1818 19 16 15+19 17 20 18 19 179 Total 38 37 37 34 32 38 35 39 37 38 365 Control-2 TalRe D3icate number Total S 1 2_

3 4

5 1 6 7

8 19 1

10 1

0 1 0

0 0

2 0

0 0

3 0

0 0

4 4

5-5 5

6 4

4 4

45 5

10 12 13 12 10 10 10 13 11 12 113 6

0 0

0 7

16 17 16 114 16 15 118 14 14 15 155 Total 30 34 134 131 132 129 132 131 129 131 313A" 100% Intake DyRepDlicate number Total Ia 2_ _

4 5

6 7 18 19 1101 1

0 0

0 2

0 0L 0

0 0

3 0

0 0

-4 6

4 6

5 5

7 5

5 4

6 53 5

11 12 12 r13 10 14 12 12 13 11 120 6

0 0 I0 0

0 0

0.

0 0

7 1_6 p C-* 15 1 1 US 1* 15 M 16 S 1s 19 1 17 1 159 Total 1I3 3

313 213 41 33 File: sqnlO1_O81412data.xlsx Entered by. J. Sumner Reviewed by V

0 Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated August 14-21, 2012 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:

8207 Concentration Replicate number Survival Average reproduction Coefficient of Percent reduction from

(%)

1 2

3 4

5 6

7 8

9 10

(%)

(offspring/female) variation (%)

control (%)

Control - 1 30 33 29 33 31 28 34 32 30 32 100 31.2 6.2 Not applicable 10.8%

36 34 33 33 35 35 34 34 31 32 100 33.7 4.4

-8.0 21.6%

36 31 33 31 30 37 35 33 32 30 100 32.8 7.6

-5.1 43.2%

37 34 34 31 31 31 37 34 34 35 100 33.8 6.7

-8.3 86.4%

34 34 36 34 35 33 34 35 36 37 100 34.8 3.5

-11.5 100%

38 37 37 34 32 38 35 39 37 38 100 36.5 6.0

-17.0 Control - 2 30 34 34 31 32 29 32 31 29 31 100 31.3 5.6 Not applicable 100% Intake 33 29 33 33 33 36 33 32 36 34 100 33.2 6.0

-6.1 Outfall 101:

Dunnett's MSD value:

PMSD:

Intake:

Dunnett's MSD value:

PMSD:

2.021 6.5 1.459 4.7 MSD =

Minimum Significant Difference PMSD =

Percent Minimum Significant Difference PMSD is a measure oftest 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 (90"h 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, 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 ofEPA 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, OIL File: sqn101_081412data.xlsx Table populated from associated "Verification of Ceriodaphnia Reproduction Totals" spreadsheet.

Spreadsheet entered by: J. Surpner Reviewed by:

7

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated August 14-21, 2012

ET Statistical Analyses Ceriodaphnla Survival and Reproduction Test-Reproduction Start Date:

811412012 Test ID: CdFRCR Sample ID:

TVA / SQN 101 End Date:

8/21/2012 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species:

CD-Ceriodaphnla dubia Comments:

Non-treated Conc-%

1 2

3 4

6 6

7 8

9 10 Control-1 30.000 33.000 29.000 33.000 31.000 28.000 34.000 32.000 30.000 32.000 Control-2 30.000 34.000 34.000 31.000 32.000 29.000 32.000 31.000 29.000 31.000 10.8 36.000 34.000 33.000 33.000 35.000 35.000 34.000 34.000 31.000 32.000 21.6 36.000 31.000 33.000 31.000 30.000 37.000 35.000 33.000 32.000 30.000 43.2 37.000 34.000 34.000 31.000 31.000 31.000 37.000 34.000 34.000 35.000 86.4 34.000 34.000 36.000 34.000 35.000 33.000 34.000 35.000 36.000 37.000 100 38.000 37.000 37.000 34.000 32.000 38.000 35.000 39.000 37.000 38.000 Intake 33.000 29.000 33.000 33.000 33.000 36.000 33.000 32.000 36.000 34.000 Transform: Untransformed Rank 1-Tailed Isotonic Conc-%

Mean N-Mean Mean Min Max CVQ/%

N Sum Critical Mean N-Mean Control-1 31.200 0.9968 31.200 28.000 34.000 6.193 10 33.800 1.0000 Control-2 31.300 1.0000 31.300 29.000 34.000 5.645 10 10.8 33.700 1.0767 33.700 31.000 36.000 4.435 10 140.00 75.00 33.800 1.0000 21.6 32.800 1.0479 32.800 30.000 37.000 7.578 10 122.00 75.00 33.800 1.0000 43.2 33.800 1.0799 33.800 31.000 37.000 6.660 10 136.50 75.00 33.800 1.0000 86.4 34.800 1.1118 34.800 33.000 37.000 3.532 10 151.00 75.00 33.800 1.0000 100 36.500 1.1861 36.500 32.000 39.000 5.954 10 150.50 75.00 33.800 1.0000 Intake 33.200 1.0807 33.200 29.000 36.000 5.991 10 Auxillary Tests Statistic Critical Skew Kurt Kolmogorov D Test Indicates non-normal distribution (p <= 0.05) 0.98421 0.895

-0.0927

-0.4597 Bartiletts Test indicates equal variances (p = 0.35) 6.58769 15.0863 The control means are not significantly different (p = 0.91) 0.12078 2.10092 Hypothesis Test (1-tall 0.06)

NOEC LOEC ChV TU Steel's Many-One Rank Test 100

>100 1

Treatments vs Control-1 Linear Interpolation (200 Resamples)

Point SD 96% CL Skew ICO5

>100 IC10

>100 IC15

>100 1.0 iC20

>100 0.9 IC25

>100 0.8 IC40

>100 0.7 IC50

>100 0.6 0.5 c0.4 10.3 0.2 0.1 0.0

-0.1

-0.2

-0.31 0

50 100 150 Dose %

Dose-Response Plot 45..

40 35 330 S25.

S20

= 15, 10l 5.

u g

N, 8

Cil C;

o~

Co (tage 48 of 99 File zqn101 081412dmetxte Entered by., SJ ter Reviewed by. -- F

TVA / Sequoyah Nuclear Plant, Outfall 101

. Non-treated August 14-21, 2012 0.:T SEnvinonn,.ntal Tentlfg Solutions.

Ins.

Statistical Analyses Analysis used for PMSD calculation only.

Ceriodaphnia Survival and Reproduction Test-Reproduction Start Date:

8/14/2012 Test ID: CdFRCR Sample ID:

TVA/SQN 101 End Date:

8/21/2012 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species:

CD-Ceriodaphnia dubia Comments:

Non-treated Conc-%

1 2

3 4

6 6

7 8

9 10 Control-1 30.000 33.000 29.000 33.000 31.000 28.000 34.000 32.000 30.000 32.000 Control-2 30.000 34.000 34.000 31.000 32.000 29.000 32.000 31.000 29.000 31.000 10.8 36.000 34.000 33.000 33.000 35.000 35.000 34.000 34.000 31.000 32.000 21.6 36.000 31.000 33.000 31.000 30.000 37.000 35.000 33.000 32.000 30.000 43.2 37.000 34.000 34.000 31.000 31.000 31.000 37.000 34.000 34.000 35.000 86.4 34.000 34.000 36.000 34.000 35.000 33.000 34.000 35.000 36.000 37.000 100 38.000 37.000 37.000 34.000 32.000 38.000 35.000 39.000 37.000 38.000 Intake 33.000 29.000 33.000 33.000 33.000 36.000 33.000 32.000 36.000 34.000 Transform: Untransformed 1-Tailed Conc-%

Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD Control-1 31.200 0.9968 31.200 28.000 34.000 6.193 10 Control-2 31.300 1.0000 31.300 29.000 34.000 5.645 10 10.8 33.700 1.0767 33.700 31.000 36.000 4.435 10

-2.828

2.287 2.021 21.6 32.800 1.0479 32.800 30.000 37.000 7.578 10

-1.810 2.287 2.021 43.2 33:800 1.0799 33.800 31.000 37.000 6.660 10

-2.941 2.287 2.021 86.4 34.800 1.1118 34.800 33.000 37.000 3.532 10

-4.072 2.287 2.021 100 36.500 1.1661 36.500 32.000 39.000 5.954 10

-5.995 2.287 2.021 Intake 33.200 1.0607 33.200 29.000 36.000 5.991 10 Auxiliary Tests Statistic Critical Skew Kurt Kolmogorov D Test indicates non-normal distribution (p <= 0.05) 0.98421 0.895

-0.0927 -0.4597 Bartlett's Test indicates equal variances (p = 0.35) 5.58769 15.0863 The control means are not significantly different (p = 0.91) 0.12078 2.10092 Hypothesis Test (1-tall, 0.05)

NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnett's Test 100

>100 1

2.02145 0.06479 32.12 3.90741 7.9E-06 5, 54 Treatments vs Control-1 I

age 49 of 99 File: sqnlOl 081412data.:dsx Entered by: J. Sumpner Reviewed by: --*7

TVA / Sequoyah Nuclear Plant, Intake Non-treated August 14-21, 2012 a

ETS Statistical Analyses

Ceriodaphnia Survival and Reproduction Test-Reproduction Start Date:

8/14/2012 Test ID: CdFRCR Sample ID:

TVA / SQN 101 - Intake End Date:

8/2112012 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species:

CD-Ceriodaphnia dubia Comments:

Non-treated Conc-%

1 2

3 4

5 6

7 8

9 10 Control-1 30.000 33.000 29.000 33.000 31.000 28.000 34.000 32.000 30.000 32.000 Control-2 30.000 34.000 34.000 31.000 32.000 29.000 32.000 31.000 29.000 31.000 10.8 36.000 34.000 33.000 33.000 35.000 35.000 34.000 34.000 31.000 32.000 21,6 36.000 31.000 33.000 31.000 30.000 37.000 35.000 33.000 32.000 30.000 43,2 37.000 34.000 34.000 31.000 31.000 31.000 37.000 34.000 34.000 35.000 86A 34.000 34.000 36.000 34.000 35.000 33.000 34.000 35.000 36.000 37.000 100 38.000 37.000 37.000 34.000 32.000 38.000 35.000 39.000 37.000 38.000 Intake 33.000 29.000 33.000 33.000 33.000 36.000 33.000 32.000 36.000 34.000 Transform: Untransformed 1-Tailed Conc-%

Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD Control-I 31.200 0.9968 31.200 28.000 34.000 6.193 10 Control-2 31.300 1.0000 31.300 29.000 34.000 5.645 10 10.8 33.700 1.0767 33.700 31.000 36.000 4.435 10 21.6 32.800 1.0479 32.800 30.000 37.000 7.578 10 43.2 33.800 1.0799 33.800 31.000 37.000 6.660 10 86.4 34.800 1.1118 34.800 33.000 37.000 3.532 10 100 36.500 1.1661 36.500 32.000 39.000 5.954 10 Intake 33.200 1.0607 33.200 29.000 36.000 5.991 10

-2.258 1.734 1.459 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.05) 0.91798 0.905

-0.1715 0.33113 F-Test indicates equalvariances (p.= 0.73) 1.2669 6.54109 The control means are not significantly different (p = 0.91) 0.12078 2.10092 Hypothesis Test (1-tall, 0.05)

MSDu MSDp MSB MSE F-Prob df Homoscedastic t Test indicates no significant differences 1.45886 0.04661 18.05 3.53889 0.03657 1,118 Treatments vs Control-2 Dose-Response Plot

'1" 40 35 30 25 L 20 15 10 5

.0 4

1-tail, 0.05 level of significance C?

8 C14 COe4 2

J9.G=

age 50 of 99 File: sqnl 01_0814 2data.xlsx Entered by: J. Sumnner Reviewed by:

ETS 6E~M-MITngSwmonth.,

Page 6of 7 Species: Ceriodaphnia dubia Client: TVA / Seauoyah Nuclear Plant, Outfall 101 Dailv Chemistr:

Date:

0 1 t-I+t..-

I.

Day

',performed pH D.O. and conductivity measurements only.)

(Analyst identified for each de 0

1 I

2 Analyst Ij~

ITS

~

SV Concentration jParameter pH (S.U.)

-?.CC II 7. s 7.143 11 51 1 '7.55 I

w DO (mg/L)

CONTROL Conductivity (ginhos/cm)

Alkalinity I (mg CaCr 3/L)

Hardness (Mng CaCO3/L)

'36C 313 1

at.

'a.

.0 S

-Y.CIL 07.q4 0

.4 7508

(

E

~46 S

Temperature (QC)

U DH (S.U.)

DO (me*J "7

.II 10.8%

Conductivity (gimhos/cm)

.1 0,L. It

-- 7

TemDerature (00)

M-4.1 a

a ~

I--*-

II I

II.

.pH (S.U.)

?7.C=

-7.qi B 2S7, S17.Si DO (mfiL) 21.6%

Conductivity (iunhos/cm)I

-7.17 1 73

-Loa

-LA-1

_15.

-1.

'7.4 9 SW~

Temperature (PC)

S.

oH (S.U.)

a -~-------4I L

43.2%

i-.-.-..--,

DO (mulL) 7.7 Conductivitk f

Sq (Aihmos/cm)

ZL 9.s; 1-55

Temnerature (C*

IlA-1

.4 - V.

14-

-1 1-L--

pH (S.U.)

7.60

-7~

7.46

]

S DO~g/)

7A' 7.__

86.4%

Conductivity Tempoerature C0C)

IN t

pH (S.U.)

-7.qq 7.47-

-1.C3 1.0, DO (mnwU 100%

Conductivity (Iunhos/cm) I

Alkalinity (Mg CaCO3/L)

T

.*Hardness (MigCaCO 3/Q)1*.

choie m1 0.1M

Temperature (00)

PH (S.U."

DO im 7.1 (pinhos/cin)

UIS 100%

Alkalinity Intake mg

Hardness

TR chlorine (mg/L) 0.00

- Temperature CC) 2S.0 15.1 R".I 1.t Initial Final Initial Final Initial Final

'Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1. Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by: __

SOP ATI I - Exhibit ATi 1.2, revision 07-01-12 R.C.wby g~0 1 K-rn ypge 51 of 99

~*

ETS Page 7 of 7 Species: Ceriodaphnia dubia Client: TVA / Seauovah Nuclear Plant. Outfall 101 Date:

O0.IL 1...

A*I:"*

Dae Ot I

L I

Day (Analyst identified for each day. performed pHi D.O. and conductivity measurements only.)

I V

Analyst I Yvj,,-.11 S Ic31 f3Z.

11 43 Concen-

. tration Parameter

-I PH (S.U.)

DO (mg/L)

Conductivity (umhos/cmr) 7A2 CONTROL

Alkalinityt (mg CaCO3/L)

Hardness (mg CaCO3/L)____

315 7AM 310

-sZ3

Temperature (QC)

pm

.7 1Lý t DH (S.U.)

'JA~.

10.8%

Conductivity I:2e 71S 65_

(iimhoslcm) v r

m

TemDerature (T)

-- A.Ve L.4~

I "..m..t PH (s.u.)

7.O7.S?

-7.94

-7.5Z DO (mg/L) 7A Ts C

21.6%

Conductivity

-2 (pmhos/cmn)

Z

______*Temperature (IC)

ir3,

'L0 LAt k s

I0

-u.. -L f

~

L.

PH (s.u.)

67.51

.S'-

7.1 7

7. SC 73 DO mglL) 7!

7

.0

7. V 43.2%

Conductivity (grnhos/cmn)

MCA V40 7,4

_____*Temperature (C) it~

1.'

-1 24 tC

. g

z vH (S.U.)

'4.F* 1-7.41 1 Z-19 11 -7.S3 1 -1.4 Li 1 7.SS.

A 7.93 V -7.22 DO (mg/L) 86.4%

Conductivity (gmnhos/cm) 07 1

13.01F th 7-4

TeM "erature (C

i S

pH (S.U.)

12q'

-1.6

-7~

76-54 t7Z DO (mc/L)

I.

I I

I 00%

I.

Conductivity

Alkalinity

Hardness

TR chlorine (mg/L)

Temperature ('C) a a-100%

Intake PH (S.U.)

DO (migUL Conductivity (gmhos/mn)

Alkalinlty (mg CaCO3QL

ardness (Mg CaCO0/L)

TR chlorine (mg/L)

Temperature (QC

_ ntil Fna niil Final IInitial Final IInitial 11Final a

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1. Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by:

SOP ATI 1 - Exhibit ATi11.2, revision 07-01-12 m*

'-"4

'-'4-'?

r__

ge 52 of 99

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated August 14-21, 2012 "E T S Ceriodaphnia dubia Chronic Whole Effluent Toxicity Test

0EPA-821-R-02-013, Method 1002.0 (1 0 0@

Daily Chemical Analyses to 0EnTs (a Environmental Testing Solutions. Inc.

Project number:

8207 Concentration IParameter Da 0 Da7 Day2 Da 3 Initial _7 Final IInitial I Final IInitial_

2 Final 1Initial Ia

.9 oH (SU) rDO (mffIL)

Conductivity (pnhoslcm)

I Control Alkalinity (maLL CaCO) 9--

310 62 88 24.9 Hardness (mg/L CaCO3)

Temperature (C) 24.71 25.11 24.91 25.0 25.21 24.91 25.2 9 -

9*-

9~

-, ~9

.9-

~.9 9-I -

~9 -

I -

pH (SU) 7.621 7.441 7.441

' 7.58 7.451 7.591 7.571 7.58 I

.. J A

A DO Ima/l,*

7-7 7-K K'O KO 70 N11 Nf0

70 10.8%

DO Arm-/T I 77 78 80 so 79 8 1 80 79 Conductivity (ILmhos/cm) 2970M 294 1_*Mff=

294 L%ý

ý 285 2 8 5 Temnerature (0I0 24.71 24.91 24.91 25.2 24.91 25.01 25_oi 25.11 24R 25

11 24~

94121 9421 250 pH (SU) 7.62 7.43 7.44 7.59 7.51 7.59 7.59 7.60 7.52 7.54 7.52 7.56 7.55 7.30 21.6%

DO(mg/L) 77 78 80 81 79 80 79 79 79 81 80 79 7.8 8.2 Conductivity (pnhos/cm) 280 283 4

280 273 272 273 ?

_____28 Temperature(70 247 249 249 250 249 250 250 251 248 250 247 248 24.8 25.0 pH(SUJ) 7.61 7.44 7.45 7.61 7.51 7.59 7.59 7.58 7.51 7.54 7.51 7.56 7.56.

7.30 4.2%

DO(mgL) 7.7 7.9 8.0 8.1.

8.0 8.0 7.9 7.9 7.9 8.1 8.0 7.9 7.8 8.2 Conductivity (pmhos/cm) 254 r 255 @

W 255 249 9 248 7r 246 259 Temperature CC) 247 24.8 25.0 24.9 25.0 25.0 25.0 24.8 24.8 24.8 24.7 24.8 24.8 25.3 PH (SU) 7.60 7.43 7.45 7.61 7.51 7.58 7.58 7.61 7.45 7.53 7.45 7.55 7.53 7.28 8.% DO (mg/L) 7.6 7.9 8.0 8.2 8.0

.8.0 8.0 7.9 7.8 8.1 8.0 7.9 7.8 8.2 Conductivity (pnhos/em) 203 203 194 192 196 196 205 Temperature (oC) 248 25.0 2.0 24.9 25.0 25.1 25.1 251 24.8 24.8 247 249 24.8 25.2 pH (SU)

!7.60 7.44 7.42 7.63 7.53 7.57 7.57 7.61 7.44 7.54 7.44 7.55 7.54 7.29 DO (mz/L) 7.7 7.9 806 8.2 8.1 7.9 8.0 7.9 7.8 8.1 8.0 7.9 7.8 8.3 Conductivity (wnhos/cm) 183 183 186 181 179 183 100%

Alkalinity (mg/L CaCO3) 72 Hardness (mg/L CaCO3 )

72 71 74 Total Residual Chlorine U

m<0.10

<0.10

<0.10 Temperature (00 24.9 25.0 25.0 24.9 25.0 24.9 25.1 24.8 25.0 24.8 24.8 25.0 24.9 25.2 pH (SU) 7.61 7.45 7.44 7.64 7.52 7.57 7.58 7.58 7.43 7.56 7.43 7.56 7.54 -

7.30 DO (mg/L) 7.7 8.11 8.0 8.2 8.2 8.0 8.1 8.0 7.9 8.2 8.0 8.0 7.8.

8.2 Conductivity (pmhos/cm) 183 186 185 182 181 176 1%

184 100% Intake Alkalinity (mg/L CaCO3) 68 687 Hardness (mg/L CaCO3 )

72 74 72!

I jKesidual Unlorine jmtL)

< 0A0

<0.10 l

<0.101 Temperature (C) 25.0 24.91 25.01 25.11 251 24.9

2.

24.71 24.91 24.7

24.

24.7 24.94 25.1 File: sqn101_081412chem.xls Entered by: J. Sumner Reviewed by: 4

I I

Ii I

I

ETS a~-~

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

Species: Pimephales promelas Client: Tennessee Valley Authority Facility: Sequoyah Nuclear Plant NPDES #: TN0020168 Project #:

g14"I County: Rhea Outfall: 101 Dilution prep 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:

-fz 0,

oL. oLb Randomizing template:

t Date and times organisms 01 t I l-.

1 C)

Incubator number and were born between:

shelf location:

Organism source:

ATby,. 60 pTCI

., oC 1i3--L Artemia CHM number:

f.1 Drying information for weight determination:

Transfer bowl information:

pH= 'Wo.tO S.U.

Date / Time in oven:

vt',

n-t'.oo Temperature = "l..

0C Initial oven temperature:

10C Average transfer volume:

Date / Time out of oven:

o0"J.. t*,

0'

""'"~

Final oven temperature:

Total drying time:

-1A 46 Daily feeding and renewal informiation:

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

O-15.101-1StL co I's%%4 L

% r.

ilt9.01 to L0IS.I L

%-..%I 3

61.11.1-1.

co-t)

I t'tQ

%UlsI?-~t.

16 k 'Loftm. II L

0-1+1E]

4 af-kt" 1L-o*1t iq%

.L.

ll.

170011. 0e 1208T 1+/-a1.0 5

tA d1Si 9 11 A

II2091-.O0 I ti20ai.ol 6

i OU'1-nan 71 Control information:

Acceptance criteria Summary of test endpoints:

% Mortality:

-<20%

7-day LCso

) I w7.

Average weight per initial larvae:

NOEC W

1 7.

Average weight per surviving larvae:

O. S l

>0.25mglarvae LOEC I tOol.

ChV 1*0 1067.

IC 2 5 5

(

1007.

age 54 of 99 SOP AT2O - Exhibit AT20.3, revision 07-01-12

Page 2 of 6 Species: Pimephales promelas Client: TVA / Sequoyah Nuclear Plant, Outfail 101, UV-treated Date:

o atwi-IM-Survival and Growth Data Day CONTROL 10.8%

21.6%

,_A B

C D

E F

G H

I J

K L

01

) It to to it) 0 t /C0t I t O C to) to 0

1 o

10 3 10 U

1O

/0

/0 10 /01 t0 2

I0 10 o

to 10 110

/0

/)0 10 3

/0 b 0 103 to 0

/0 10 10 10 IC 10 4

1C)

IQ C 10

/o (0 10/0 toIC to0

/0

. lo 1

o

/0 /0_i /00IC 1c M

Q 6

10 101(0 to/0 /0 (0 /0 10 (0 '0 /0 7

IlV

/*

10 to, 0

3 10 0 0

to

/0' (0 A = Pan weight (mg)

Tray color code::

Analyst:

146 1--..

s 1-3 t4.4z 4m.v-8 14.7%{,

As

3.

.4.l4.

orp Is.

Date:

08.06.1t-B = Pan + Larvae weight (mg)

Analyst:.

1`.

I".i6 aq.zs tl.36

.Z 1

2Loo06 2.o.21 Zas57 IS.1 dt Z0.34 l.qs q.5$5 1-113 Date:

op.z.7. -I C - Larvae weight (mg) = B - A Hand calculated.

S tO S

A

'sC

(-1 S.S 6,O

.'b% Is N4 A s.'Il S.aIi

.0-Analyst:,_

Weight per initial number of larvae (mg)

= C / Initial number of larvae Hand calculated.

.1 Analyst:

I y

N 49 0*

0

'9

'0 0*

4 0'

Average weight per Percent reduction initial number of from control (0le) o.6'!S U -

U -

- tqA7.

a i

-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.

(Iage 55 of 99 SOP AT20 - Exhibit AT20.3, revision 07-01-12

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

OI.i'A-VL.

Survival and Growth Data Day 43.2%

86.4%

100%

,_M N

0 P

Q R

S T

U V

W, X

0 0Q 1D Ito I ri to to 10 C) t) to

/0 2

00 10 10 (C 10 (0 10 to i0 to1

[ 0 to I_ Q10 101 0

10 IO

)

1 to 1O 0

to00 3

I to I to 10O

( to /

0 10 I

/0 /0

/ ) / 0 4

I)

/0 10

/C)

/0

/

0 ItI 10

/0

/0 IC) /I

)

5

/0 /0 10 /0 10

/0 /0

,1A /01

/0

/0 6

to to 10 to to to 1

0t 11

/0 t O1 0 77__

(o 10 t0

1) t 01 1

I*

/0

/b

/6Q A = Pan weight (mg)

Tray color code::

L

-L&,L 1l.-

Analyst-In 34D I.'

13.*.14 135.77' 15.A 13-1&*

14.8z

,4.3s IL-.3 13.Uf ILI.

Date: _

_n_..__-

B = Pan + Larvae weight (mg)

Analyst:

-MR 7-_.oo 1L63 1_0I I41.041 18_.36

_._6

_q.47 tq___

16.1_ IS 17._7_

Date:

o8.zI7 C = Larvae weight (mg) = B - A Hand calculated.

SA.'L S~iI. 1 6

t S.t t

S.d% S.A-S..z% 4.1t S S60 SL.6 7

2.J,

.sc'i Analyst:

Weight per initial number of larvae (mg) 1 C/Inltial number of larvae Hand calculated.

V,%*

i e

Analyst.

C o"

v" U"

0" L Average weight per Percent reduction initial number of from control (%)

o--.)7.

.¢OP.

q..&T.

0, S

.- ).

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

Comments:

(f age 56 of 99 SOP AT20 - Exhibit AT2O.3, revision 07-01-12

Page 4 of 6 Species: Pimephales promelas Client: TVA / Seciuovah Nuclear Plant. Outfall 101. UV-treated Date:

O*.L M I

I I

Day 100% Intake Y

Z AA BB 0

to10 10

(

S/00 2

to ot 3

I b t o t o t

4

/0 to (0 10 5

1

_)/0 t

0 6

(0 (0 t0 10 7

/

i 10 to, (0

A = Pan weight (71g*

Tray color code::

4 Analyst:

lit.

i.1 Date:

O_.__

B = Pan + Larvae weight (nag)

Analyst-J-r 2

Z

.3 Date:

0O.z7.1z C = Larvae weight (mg) = B - A Hand calculated.

(o L' L Analyst-Weight per initial number of larvae (mg)

= C./ Initial number of larvae Hand calculated.

Average weight per I Percent reduction initial number of from control larvae (mig)

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.

(Iage 57 of 99 SOP AT20 - Exhibit AT20.3, revision 07-01-12.

(D Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 UV-treated August 14-21, 2012 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.

8207 Not for Compliance Assessment, Internal Lablratory QC Conenatratio()

Repfle letlalauamberf mI al-mber-aF an. A=P-uw eght(mg)

B-Paa+Lar-se Larewelght(mg)

Weight/SonIhag Me-aweight/

Coeffictafvarlatlon Weight/lldtd lnumber Meanerrival Merwreghtl/talt CeltTdeletof Peent reductonfmthan larvae weightnOg)

-A-B mumberaftervue(mg)

Surtvirgotreof ateklrw -b oPSot tae(eag)

(0) humberoflarvee varitdion g

e0ovtn (00) t-r) m5.r... (%)

(ea 00 ee ra=i,,.)

A 10 10 12.88 17.98 5.10 0.510 0.510 B

10 10 14.40 19.28 4.88 0.488 0.488 Control C

10 10 15.49 21.36 5.87 0.587 0.535 8.3 0.587 100.0 0.53S 8.3 Not applicable D

10 10 14.63 20.16 5.53 0.553 0.553 E

10 10 13.33 20.06 6.73 0.673 0.673 10.8%

F 0

10 14.62 20.21 5.59 0.559 0.614 8.0 0.559 100.0 0.614 8.0

-14.9 G

10 10 14.26 20.57 6.31 0.631 0.631 H

10 10 12.84 18.78 5.94 0.594 0.594 1

10 10 14.35 20.34 5.99 0.599 0.599 2

10 10 13.19 18.43 5.24 0.524 0.524 K

10 10 14.06 19.53 5.47 0.547 6.0.547 L

10 10 13.11 18.93 5.82 0.582 0.582 M

10 10 14.08 20.00 5.92 0.592 0.592 N

10 10 13.47 18.63 5.16 0.516 0.516 432%

0 O1 35 90

.80580.561 6.4 058100.0 0.561 6.4

-5.0 0

10 10 13.53 19.01 5.48 0.548 0.548 P

10 10 13.17 19.05 5.88 0.588 0.588 Q

10 10 13.27 18.36 5.09 0.509 0.509 86.4%

R 10 10 13.69 18.96 5.27 0.527 0.521 4.5 0.527 975 0.509

&.6 4.8 S

10 10 13.96 19.47 5.51 0.551 0.551 T

10 9

14.82 19.30 4.48 0.498 0.448 U

10 10 14.33 19.83 5.50 0.550 0.550 V

10 10 12.73 18.38 5.65 0.565 0.565 100%

V ot 27 83

.50550.534 5.8 O5 100.0 0-534 5.8 0.1 W

10 10 13.24 18.50 5.26 0.526 0.526 X

10 10 12.76 17.70 4.94 0.494 0.494 Y

10 10 15.22 21.43 6.21 0.621 0.621 100% Intake Z

0 10 14.61 20.34 5.73 0.573 0.573 100.0 0.581 12.5

-8.7 AA 110 14.37 20.84 6.47 0.647 0.647 BB 10 10 14.39 19.21 4.82 0.482 10.482 Outfafl 101:

Dunnett's MSD value:

PMSD:

Intake:

Dunuett's MSD value:

PMSD:

0.0685 12.8 0.0826 15.5 MSD =

Minimum Significant Difference PMSD =

Percent Mirinium Significant Difference PMSD is a mensum 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 deter-mined by USEPA (I 0th 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 Interiaboratory Variability Study (USEPA, 2001 a; USEPA, 2001 b).

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, O1.

File: sqnl01 081412data-uv xlsx Entered by: J. Sumner Reviewed by:

TVA / Sequoyah Nuclear Plant, Outfall 101 UV-treated E

S August 14-21, 2012

.Statistical Analyses EFIonIt frgmetlTl*SolutO n sfle.

Larval Fish Growth and Survival Test-7 Day Growth Start Date:

8/14/2012 Test ID:

PpFRCR Sample ID:

TVAI SQN 101 End Date:

8/2112012 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species:

PP-Pimephales promelas Comments:

UV-treated Conc-%

1 2

3 4

D-Control 0.5100 0.4880 0.5870 0.5530 10.8 0.6730 0.5590 0.6310 0.5940 21.6 0.5990 0.5240 0.5470 0.5820 43.2 0.5920 0.5160 0.5480 0.5880 86.4 0.5090 0.5270 0.5510 0.4480 100 0.5500 0.5650 0.5260 0.4940 Intake 0.6210 0.5730 0.6470 0.4820 Transform: Untransformed 1-Tailed Isotonic Cone-%

Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD Mean N-Mean D-Control 0.5345 1.0000 0.5345 0.4880 0.5870 8.269 4

0.5744 1.0000 10.8 0.6143 1.1492 0.6143 0.5590 0.6730 7.973 4

-2.807 2.410 0.0685 0.5744 1.0000 21.6 0.5630 1.0533 0.5630 0.5240 0.5990 6.009 4

-1.003 2.410 0.0685

'0.5630 0.9802 43.2 0.5610 1.0496 0.5610 0.5160 0.5920 6.414 4

-0,933 2.410 0.0685 0.5610 0.9767 86.4 0.5088 0.9518 0.5088 0.4480 0.5510 8.649 4

0,906 2.410 0.0685 0.5213 0.9075 100 -0.5338 0.9986 0.5338 0.4940 0.5650 5.806 4

0,026 2.410 0.0685 0.5213 0.9075 Intake 0.5808 1.0865 0.5808 0.4820 0.6470 12.504 4

Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.94963 0.884

-0.1675 -1.1562 Bartlett's Test indicates equal variances (p = 0.97) 0.84084 15.0863 Hypothesis Test (1-tall, 0.05)

NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnett'sTest 100

>100 1

0.06848 0.12811 0.00527 0.00161 0.02854 5, 18 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point SD 95% CL(Exp)

Skew IC05 59.875 ICIO

>100 IC15

>100 1.0 IC20

>100 0.9 IC25

>100 0.8 IC40

>100 0.7 IC50

>100 0.6 0.5 0.4 0 0.4 S0.3 j

0.2 0.1 0.0

-0.1

-0.2

-0.3 0

50 100 150 Dose %

Dose-Response Plot 0.8 0.6.

0.6 1-tail, 0.05 level of significance 00.-(:30.3 0.2 0.1 01 V

D 0

6 File: sqnl 01081412data-uv.xlsx Entered by: J. Su er Reviewed by:

Sage 59 of 99

TVA / Sequoyah Nuclear Plant, Intake UV-treated August 14-21, 2012 0:T 1

21 lfmfra Tetn Sttonj. Inc.

Statistical Analyses Larval Fish Growth and Survival Test-7 Day Growth Start Date:

8/14/2012 Test ID: PpFRCR Sample ID:

TVA/ SQN 101 - Intake End Date:

8/21/2012 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date: August 2012 Protocol: FWVCHR-EPA-821-R-02-013 Test Species:

PP-Pimephales promelas Comments:

UV-treated Conc-%

1 2

3 4

D-Control 0.5100 0.4880 0.5870 0.5530 10.8 0.6730 0.5590 0.6310 0.5940 21.6 0.5990 0.5240 0.5470 0.5820 43.2 0.5920 0.5160 0.5480 0.5880 86.4 0.5090 0.5270 0.5510 0.4480 100 0.5500 0.5650 0.5260 0.4940 Intake 0.6210 0.5730 0.6470 0.4820 Transform: Untransformed 1-Tailed Conc-%

Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD D-Control 0.5345 1.0000 0.5345 0.4880 0.5870 8.269 4

10.8 0.6143 1.1492 0.6143 0.5590 0.6730 7.973 4

21.6 0.5630 1.0533 0.5630 0.5240 0.5990 6.009 4

43.2 0.5610 1.0496 0.5610 0.5160 0.5920 6.414 4

86.4 0.5088 0.9518 0.5088 0.4480 0.5510 8.649 4

100 0.5338 0.9986 0.5338 0.4940 0.5650 5.806 4

Intake 0.5808 1.0865 0.5808 0.4820 0.6470 12.504 4

-1.088 1.943 0.0826 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.95559 0.749

-0.6317

-0.2536 F-Test indicates equal variances (p = 0.44) 2.69933 47.4683 Hypothesis Test (1-tail, 0.05)

MSDu MSDp MSB MSE F-Prob df Homoscedastic t Test indicates no significant differences 0.0826 0,15453 0.00428 0.00361 0.31833 1, 6 Treatments vs O-Control.

Dose-Response Plot 0.8 0.7 0.6 f;

0..

1-I

-tail, 0.0 5 level 0.4 of significance 0.3 0.2 0.1 0

co 0

c; dz File: sqnI01_081412dEtta-uv.xlsx Entered by: J. Sumner Reviewed by age 60 of 99

I*ETS Spec Clien Dai Page 5 of 6 ies: Pmneyhales promelas it: TVA / Sequoyah Nuclear Plant, Outfall 101. UV-treated Chemnstrv:

Date:

IDay (Analyst identified for each day. performed pH. D.O. and conductivity measurements only.) 1 I,

m Analyst 0

I 1

I 2

Concentration

[Parameter I DH (S.U.)

I DO (me/L)

Conductivity (Ujmhos/cm'l CONTROL UV-treated

Alkalinity (mg CaCO3/L)

Hardness (Mg CaCO3/L)

I.,v......

m*

T emuperatureUL

a IL, 66 11~ wt -1 pH (S.U.)

7I.CC

'7'f 7.5a 1S 1C DO (mg/L) 7,_

1 7.q 10.8%

Conductivity vqzBZIS (pimho sLq 7.4

_______*Temperature C0C)

ILAI VA-I

'L -*

(

pH (S U.)

'7.CC 1-747 7.50 IZ C

DO (mgfL)

V 1

7.4 7

21.6%

Conductivity (jimhos/cm) 7-9Zg4 e

______*Temperature (C)

-L4 1.

'8 11 pH (S.U.)

7.4S 7.Llq I -i'Ll l7.C ftQ~~L DO (mg/L) 43.2%

Conductivity (nhsfcmn)

Temnlerature ('C) a a

PH (S.U.)

Z52

-8.

C3'7.Lo DO (mp/L) 86.4%

Conductivity (jpmhocm

Temperature (QC)

S S ~

~u tv ii~

PH ru.

I DO (ma/L)

I" 100%

Conductivity (iunhos/em)

-Alkalinity (mg CaC0iIL) m

Hardness S(me CaMO/L)

-I

TR chlorine (mg/L)

?ICZ

-q3 6 7..9 0

~~7.4579 1'. 0

-S.

?_.

Co7.6

.5 (O.I6 I

Temperature (OC)

DH (S.U.)

DO (ma/L)

Conductivity (pimhos/m

Alkalinity (mg CaCOs/L) 100%

Intake

Hrdness"

______I (mg CaCO3lL)

Ij

TR chlorine (mg/L)

-(o.

Temperature (Q "t... IS 0.0

! I Final Initial Final Initial Final

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table locad onPi Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to ths be

. Total residual chlorine was performed on non-treated Outfall 101 and Intake samples.

SOP AT20 - Exhibit AT20.3, revisd:n07-1 age 61 of 99

I

&M-ei Page 6 of 6 Species: Pbnephales promelas Client: TVA / Seaunah Nuclear Plant. Outfall 101. UV-treated Date:

O&lLt.-.

Day (Analyst identified for each day, erformed PK DO. and conductivi measurements only.

3 4

5 Concer-Parameter trati0n pH (S.U.)

-,7 1.S, 7.0 7."

77.

7 Conductivity CONTROL (jirnhoslcm)

UV-treated

Alkalinity (mg CaC03/L

Hardness (Mg CaCO3/L)

2.

IA-.

2.4-i

_________*Temperature (O*

PH (s.u.)

7i

.8 74

.67 7.7 C1(

.3 DO (mnW

-7.

I I

K V

10.8%

Conductivity li.

TemperatureC)

I S.O C

n.

a..

4.1 "M4 PH, U.)

7.

_ o___

C

.3 DO (mgpL) 0

-7.

1 21.6%

Conductivity (11mhos/cm)2-p s

-_*Temperature (Q)

11.

IIs'.

o I.

I pH(S.U.)

-7 54 DO (mg/L).'8" 7.9 7..I9.0 1.9 7.9...

43.2%

Conductivity (gxmbos/cmn)

LC-L17

_____*Temperature (OC)

Is -

L &

~

~U.0 if'.

PH (S.U.)

7.

7.603

.*I 1

7.

7.3 DO hlorin1

.mA 7.j.

86.4%

Conductivity

Temperature CC)

LS..

I.S__0

2.

V.t 7_ S-.0 Conductivity

-Ios/cm 167 1%

Alkalinity 1

(%mg CaCO3/L 0

Hardness

TR chlorine (rag/L)*

- Temperature (IQ)

"I-%.

2LS PH (S.U.)

7.47LI i~

7A.2 sq 7.

DO mglL)

I 7..0.97.

dq Conductivity Whos/cm) 176 (Sol 100%

Alkalinity10 Intake (mg CaCOs/

Hardness (Mg CaCO31L)1-

TR chlorine (mg&L) 0

____*Temperature

Q IS.

1.(.f 2q.

11.Z S I

I

.7 s

Initial Final Initial 11 Final I

Initial 11 Final I

Initial I

Final

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1.

Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bcnch sheet Total residual chlorine was performed on non-treated OutflI 101 and Intake samples.

SOP AT220- Exhibit AT20.3, revision 07-01-12.

\\*jP~yage 62 of 99

0 cc Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 - UV-treated August 14-21, 2012 Pimephalespromelas Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Daily Chemical Analyses Project number:

8207 Concentration Parameter Da 0 Day I Day 2 Da Da y4 Day 5 Day 6 Initial Final Initial Final Initial Final Initial Final Initial Final Initial.

Final Initial Final PH (SU) 7.65 7.49 7.50 7.59 7.62 7.53 7.59 7.47 7.56 7.40 7.59 7.61 7.62 7.34 DO (rneIL) 7.8 7.8 7.9 7.8 7.9 7.9 8.0 7.8 8.0 7.6 8.1 7.9 7.9 7.8 cnrl Conductivit (Imhos/cm) 302 309 1302 303 299

,298 929 M31610 Cnrl Alkalinity (mglL CaCO 3) 62 61 61-Hardness (mg1L CaCO3) 84

.j84 RE2784

_______Temperature COC)

24.8 247 24.8 24.7 24.8 24.9 24.9 247 24.9 24.8 24.8 24.6 24.9 2.

PH (SU) 7.66 7.48 7.50 7.56 7.63 7.52 7.62 7.45 7.58 7.41 7.57 7.57 7.61 7.34 DO (mg/L) 7.9 7.8 7.9 7.8 7.9.

7.

8.0 7.8 8.01

!7.6.

7.9 7.8 7.9 7.8 0.% Conductivity (Iwnboslcm) 294 298 293__

291__

285__

282__ ffV2X_

_______Temperature (90 24.9 24.5 24.8 24.7 24.9 25.0 25.0 24.7 25.0 24.8 24.8 24.7 24.9 24.5 PH (S11) 7.66 7.47 7.50 7.52 7.62 7.52 7.63 7.45 7.56 7.46 7.57 7.58 7.61 7.34 2.% DO (mal) 7.9 7.7 7.9 7.9 7.9 7.8 8.0 7.8 7.9 7.8 8.0 7.8 7.9.

7.9 2.%

Conductivity (pmhnos/ci) 281__

284__

280_

277_27_27_28

_______Ternperature CC) 24.9 24.81 24.8 24.7 24.9

-24.8 25.1 24.8.

25.0 24.8 24.9 24.7 25.0 24.6 PH(Sim 7.66 7.45 7.49 7.54 7.62 7.52 7.62 7.45 7.55 7.411 7.54 7.58 7.61 7.33 DO (mgtL) 7.9 7.7 7.9 7.9 8.0 7.9 8.0 7.8 7.9 7.8 8.0 7.8 7.9 7.9 4.% Conductivity (pinhostcm) 255

~

258 257 255__

246__

249__

256__

_______Temperature

(*C) 24.9 24.7 24.8 24.7 24.9 24.81 25.1 24.8 25.0 24.6 24.9 24.8 25.0 24.6 PH (SU) 7.63 7.43 7.48 7.56 7.59 7.50 7.61 7.45 7.50 7.38 7.49 7.57 7.59 7.33 DO (nmg/L) 7.9 7.7.

7.9 7.9 8.1 7.9 8.1 7.7 8.0 7.5 7.9 7.8 7.9 7.9 84%

Conductivity (ismhos/cm) 204 2JM

=07 20'7 ~.

'204 197 98_

-207__

_______Temnperature (0C) 24.9 24.7 24.9 24.7 24.9 24.8 25.1 26 25.0 24.71 24.9 24.8 25.0 24.3 PH (SI1) 7.62 7.43 7.48 7.54 7.591 7.49 7.61 7.2 7.50 7.41 7.47 7.57.59 7.31F DO (mWL 8.0 7.7 7.8 7.9 8.31 7.9 8.1 788.1 7.5 8.0 788.0 7.9 Conductivity (gmhos/em)

16188, 18'6 182 178 181

.187

[N M

100%

Alkalinity (mg/L CaCO3) 70 70

'70 Hardness (mgfL CaCO3) 72 72 72

Total Residual Chlorine (mg/L)

<010

<0.10<01

_______Temperature COC) 25.0 246 25.0 24.6 25.1 24.7 25.1 24.6 25.1 24.7 25.0 24.7 25.1 24.2 PH (SlY) 7.60 7.42 7.48 7.55 7.57 7.53 7.61 7.43 7.47 7.42 7.461 7.58 7.59 7.32 DO (mg%)

.8.10 7.7 8.0 7.9 8.2 8.0 8.1 7.8 8.1 7.7 8.0 7.9 7.9 8.0 Conductivit (Iwhos/cm) 186

-IO 191 184 182 &

.YJ M

178 178 189 MWMN 100% Intake Alkalinity (mgfL CaCO3) 70' 70 7

Hardnesa (mg/L CaCO3) 71 72 7

ýTotai Residuai Chiorine 'es.1w 1W.1

<01

<0. 10 Temp erature (0) 25.1

2.

250 24.61 25.0 24.9 25.1 24.61 24.9 2.48 2.512.

NOte:.otal residual chlorie was performed on non-treated Outlall 10l and Intake samples.

File: sqnl01_081412chem-UV.xls Entered by: J. Sumner Reviewed by:

b

.ETS SEtwimnmentBITesting Solutions, Inc.

Page -7 Page I

of 3 Time initiated 15 0, Time completed ict Analyst

]

Date analyzed Ei.iK.ZZ Thtrant normalitv and multinlier detennin atlos Alkalinity (SM 2320 B)

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

a no and T...

p11 of Normality Normality (N) of H2SO4 pH Factor or Multiplier Deionized Titrant check Begin End Total f (5 ml Na2CO3 x 0.05)/E (N x 50000)/ 100 ml sample water reference standard ml ml ml

= 0.25/E

= Nx 500

= 4.5 S.U.

number number (E)

(acceptable range = 0.0180 -0.0220) 5.1

%0%q~

494 it oNSo 10119 0.0

~

o 1o.3

-1k tote: A*-

Laboratory control standard:.

Reference standard True value Sample Alkalinity (MV)

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

(acceptable range (Ing CaCO31L)

(ml) ml ml ml

=90 to 1!0%)

io3t" 100 100 it. I tl.*

V,.9 10.3 9q 1.90,%

Duplicate saf pie precision:

Sample Alkalinity

%RPI) =

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3IL)

((S - D) /[(S+r)g2-) x 100 number (ml) ml ml ml (acceptable range=.+ 10%)

Duplicate (B)

D Matrix spike recovery:

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

(mg CaCO3IL)

(mi) ml ml ml

-fN$I tO37 sO I0L>

Z7.7

-$3.S 16.g (o.3 Iio 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%)

'-19

¶ :oL.

Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID (ml) ml ml ml Multiplier (Ing CaCOL) 00 a

04"AL H

lSLO t 30,S 14.S 6.0 103 (OZ

.I..t

.0.0 6.o

.o 001 GaA4i6L G.)

117.3 11.3 1_

"Oe.lmIffl, 17.3 LS.3 11.0 1

110

__.___r___,

Z1.3 31.t -

o.q I

U0 GO 35.1.

4S.7 C-5 CZ)

Go MAC,-

'H.

I 1

100 0.0 Co G.o 0_

Reviewed by:

I IJX Date reviewed:

00t (.

Page 64 of 99 SOP C6 -Exhibit C6.1. revision 06-01-11

.ETS 1 Endonmtnjle In Page

-7 t Page

7.

of 3 Analyst I F Date analyzed Iog.vI.]t 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 normalltv and multiplier determination:

pH of Normality Normality (N) of H2S0 4 pH Factor or Multiplier DI)O"RI"T- -- Ttrant--_ft iýn End Total

= (5 ml Na2CO 3 x 0.05)/E

= (Nx 50000)/100 ml sample water reference

-standard ml

= 0f,5/E ffi Nx 500

= 4.5 S.U.

number number (E)

_0220)

Laboratory control standard:.

Reference standard True value Sample Alkalinity (MV)

%RS=MV/TVx 100 number (TV) volume Begin End Total Multiplier (mg CaCOsIL)

(acceptable range (119 CaC3/L)

(ml) ml ml ml

= 90 to 110%)

I,,

0 3-100 100 7.9 q5

.3 Duplicate sam pie precision:

Sample Alkalinity

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

{(S - D) /l(S+l)/2]) x 100 number ml ml ml (acceptable rarge = I 10%)

Duplicate (B)

D 1,1 3 A 7Z 72 Matrix spike recovery:

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

(me CaCO3/L)

(mln) mli ml ml V-15S 1037 too J

3o.9

-3q.o s.Z

) 1io.3 17o Sample alkalinity (B)

Measured spike value (MV)

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

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

-75 to 125%)

72 q.oy.

Sample measurements:

Sample volume Begin End Total Alkalinity Sample number Sample ID (ml) ml Iml ml Multiplier (me CaCO3L) ao0jt1.tO

&f/A'

)

IS. 0 nu 31.o qZ.4 3.4 ftjO1 76 I-LoS

.o&

4 LCLAI 45A 3.5 73

ot I-of "Tam st~o')

I krT

.o0 3.3 3.3 7

I't_.

l

-I I.

1_

3._

G4.6 3.3 7

I____r

_. 04 C.

6.

to.0 3.'4 70 I7,01i 1 -GI AQIiba i0 1___

10.0 11.4 3.4 70

z~o~1.O 4/13._

4.

16.1.

3.4 C~MO I~

V_____

O23.,f

%I___

7b Reviewed by:

I [

7I Date reviewed: [o et,'l.17-Page 65 of 99 SOP C6 - Exhibit C6. 1. revision 06-01-11

ETS En~viistmenitakiting Sltos n

Page 7Z Page 3

)f _.

Analyst I

Date analyzed [i.1i.1 I

Alkalinity (SM 2320 B)

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

Time initiated Time completed Z

ra 011.112.Z T7trant normallhN, and multiplier determination:

pH of Normality Normality (N) of H2SO4 pH Factor or Multiplier

"~m---T-itroat. check Begin End Total

= (5 ml Na2CO 3 x 0.05)/E

= (Nx 50000)l 100 ml sample water reference ai

ý ml ml

= 0.25/E

= N x S0

=4.5 S.U.

number number.

(acceptable range = 0.0180 - 0.0220)

Laborator control standard; Reference standard Truevalue Sample Alkalinity (MV)

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

Volume Begin End Total Multiplier (mg CaCO3/L)

(acceptable range (mg CaCO3L)

(ml) ml ml ml 90 to 110%)

-X~ss io37 100 10O Z.3. C 3.1

%.5 10.3

'"to Duplicate sam i precision:

Sample Alkalinity

[

%RPD=

Sambe S

V) volume Begin End Total Multiplier (mgCaCO (mg

-D)IRS-D)fl) x 100 Matrix spike recovr:____________

Rfrnce standard Spike value Sample LutiperSpike alkalinity (A) nubr(SV) volume Begin End Total Multiplier (mg CaCO3/L)

(rag CaCO3/L)

(ml) ml ml ml

&sS to17.

103.5 4.7

42.

e-5 \\o.

170 Sample alkalinity (B)

Measured spike value (MV)

%R=MV/SVxl0 (ra1 CaCO3/L)

= 75 to 125%)

-70.

100 Sample measurements:.

.=

1%

Sample volume Begin End Total Alkalinity Sample number Sample ID (mi) ml ml ml Multiplier (mg CaCO/L) 1S)'x RO3

_1_6 4..

1 3.,4 4t1 o

7o

a.

o I f(4Ao)

I JoQ o.,

1.0 0.9 4

r2L of 1%

-a I.o

__c 3.4 lxatR, \\

q~q 7.1 3.5 3,'C; as.....

Reviewed by:

[IZ*

I Date reviewed: F06t_,9 Page 66 of 99 SOP C6 - Exhibit C6. 1. revision 0 6-01-11

AIM SErnvronmentalTesting Solutions, Inc.

Time initiated& ".

Time completed Total Hardness (SM 2340 C)

RL = 1.0 mg CaCOA/L Analyst [3 Date analyzed [o.g.i.1 Tltrant normalitv and multiblier determination:

Titrant Normality check Begin End Total Normality (N) of EDTA.

pH Factor. or Mutiplier reference standard ml ml ml

= 0.21E

= (N x 50000)/.$1). m'inIipM'0le6 number number (E)

(acceptable range = 0.0180 - 0.0220)

-Ni 66.0 r:.K 418 Iiss 10 3 0.0 10o.

o.

o.ojq iq.c....

LaboratorY control standard:

Reference standard True value Sample Hardness (MV)

% RS=*V/ TV

100 number (TV) volume Begin End Total Multiplier (iag CaCO3/L)

(acceptable range (mag CaCO3/L)

(ml) ml ml ml

= 90 to 110%)

)oqS 40 502 l2.7L!2.7 2-I eli s,

Duplicate sam ple precision:

Sample Hardness

%RPI Sample Sample ID volume Begin End Total Multiplier (mg CaCO3lL)

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

% Z 14.-

4-7 S

14.

_---------_N oejLDI~

uplicate (B) 7 7

.D 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 9145's 1041 4o W

M~7 z3.7 C-S KC~

3 Sample hardness (B)

Measured spike value (MV)

%-R=M/SVxl10 (Iag CaCO3/L)

MV = A-B (acceptable range (Iag CaCO3/L) 75 to 125%)

Sample measurements:

Sample volume Begin End Total Hardtiess Sample number Sample ID (ml) ml ml ml Multiplier (ag CaCO3L)

TV = ND Blank (should be = 0 mg CaCO3 /L)

D 0.0 0.0 0..

ND

-11_,

-4 z3.7-7-7. '4 L.5 S.I__

)zo~s-I 7.t7.7

.37

".4

'.7 et.44m PAA 3Zi

.4 3.5 1.1 08-m12.i-4 frh%4SIL' 0

33.5 37.c Li.:s 9_

____________3?.S i Z. I 4.3s 64 o

4Z.1

'46.'4 L1.3 114 6.S~.1 1UN-4 h

O 5W got_

0.0 3.7 3.7 iZO1SIOZ._

_3.

1.3

3.

71 1_______

_____7.~

CeI k

1 11

_SI___1-.q_

741 N*geil6 OlU* is used, sample must be diluted.

Reviewed by: I e

]

Date reviewed I081+tZI nl........

L I.

EnvrIoirnfeftltTeatlngSolutIans, Inc.

Page 3q Page IZ of 3 Time initiated [7 C Time completed

-Z._

Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO3/L Analyst a Date analyzed[~ 3

]

Titrant normality and mult~ipier determination:

I Titrant Normality check Begin End Total Normality (N) of EDTA pH Factor or Multiplier reference ldll al i-

= 0.2E

-= (N x 50000)/ *50 ml sample number number rE)

(accepta 0.0220)

=Nx 11000 I

Laboratory control standard:

Reference standard True value Sample Hardness (MV)

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

(acceptable range (Ing CaCO3/L)

(ml) ml ml ml

= 90 to 110%)

VMS$ 16ki 40 50

11.

M 13.1 K.

qC "57

-6. **

Duplicate sam ple precision:

Sample Hardness

%RFD Sample Sample ID volume Begin End Total Multiplier (Ing CaCO3/L)

{(S - D) /[(S+:D)/2]) x 100 number (ml) ml ml ml s-2otio*i.-I TV SWi wT I d) 1C. 7t 32....

119 7Z.

Duplicate(B)

.'7 D

Z-Matrix spike recover:

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

(mg CaCO3sL)

I ml mi ml xj,3,ss 10q S-4c, 1C.-7 "t-ZA 5.1*

1t.C10 Sample hardness (B)

Measured spike value (MV)

%R=MV/SVx 10 (mag CaCOd/L)

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

= 75 to 125%)

"/2.9 S.; 01.0 Sample measurements:

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

TV--N-a Cs -11.t, mz.

(should be = 0 mg CaCO3/L)

_0_

.3 1-4..

41.q "5 111111111.-r 7z.

i aLoA.V,01

-TVAS JIOIUU 1 q.q 33.C 3.7 7Z.

I.39tljO 1

2-3S.C 37.'j 7%.

_-Z.O__

_O_

-3 W. 3

"'f.4 3.7 7Z

_________Z.

'44.c

~413 3.7 1

M v~a~l1.O¶ 30.0 3S.'?

3.7 vuLOSIa4.0 f

AUOV aJ 1____

3.1 1A LA' 1___t Ll I is used, sample must be diluted.

Reviewed by: "

w.

Date reviewed q ' I

]

SOP C7 - Exhibit C7. 1. revision 06-01-11

~

Envfronme~ii~sigSolutionsj nc.

Page o

Page 3

of 3 Time initiated Time completed

-'I Total Hardness (SM 2340 C)

RL = 1.0 mg CaCO 3/L Analyst nId 5ili d

Date analyzed [I *q IIIIIII Titant normnality and multipler deltermination:

Titrant Normality check Begin End Total Normality (N) of EDTA pH Factor or Multiplier erIeffft-5

,,i ml ml

= 0.2/E

= (N x 50000)l 5O ml sample number number (E)

(acceptable range -

=Nx 1000 ~1 j

Sample volume Begin End Total Hard ness Sample number Sample ID (ml) ml nml ml Multiplier (mg Ca(O0/L)

(should be = 0 mg CaCOJL)

,1.,,10.0' F.LLW.&

IL.

AD

-7.7 I1.o 3.3 Iq.C

,_s vt-o*%

t.0 J/4

-3' 4iI1.6 15.5

'is P

>4jf~l~tiW*t is used, sample must be diluted.

Reviewed by:

I]

Date reviewed

-n iII]

SOP C7 - Exhibit C7.1. revision 06-01-11

Page qz Page 1

of_

o

'PETS EnvironmentallbongSo~lgOutionls, 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

T%3 Date analyzed Oi. 1 -I.

Calibration:

I(odidereagent:

T"Nr 514C Acid reagent: I tSri I

0.10 img/L 1.00 mg/L I

Slope Reference standard number

-of 1.06

-mg/L, the saCples I

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

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 10%)

rj~ss Io*-t 0.50 o.Szq log.-

Duplicate sample precision:

Sample Sample 11D Sample characteristics Residual chlorine

%RPD = [(S - D) I[(S+D)/21) x 100 number I(MgL (acceptable range =:i: 10%)

o*'4. E PGLAhjctb S10t1I..

1d' 4, DuplicateD____

6 8 4V Sample measurements:

sample Sample 10D Sample characteristics Residual chlorine number Reagent Blank 1.-

DOI?

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

Laboratory control standard:

Reference standard Trevau (TV)

IMeasured value (MV)

% RS MV/TV x 100 number I

mg/L)

I(mg/L)

(acceptable range = 90 to 110%)

12 *.s

\\

1064 0.50 0.471 Reviedr.

.y ZII, Reviewed by

" *\\'"

]

Date reviewed SOP C8 -Exhibit C8. 1, revision 06-01-11 Page 70 of 99

Page

-tS Page of i

".ETS 6 Envlmnmental Testing Solutlons6 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 [0 Date analyzed 061.lC.

I,odide reagent: F sic Acid reagent: [

Jz L slP Calibration:

0.10 mg/L I

1.00mg/L

]

Slope I

rReference standard number

-X14SS lOC 1

0 o

-4.ocY 771 Note: For samples with a residual chlorine of > 1.0 mg/L., the samples must be diluted to be within the calibration range.

Laboratory control standard:

Reference standard True value (TV)

Measured value (MV)

%RS=MV/TVx1001 number (mg/L)

(mg/L)

(acceptable range = 90 to 110%0)

CArSs 10CL 0.50 O.53%

IoG.q]

Duplicate sample precision:

Sample Sample ID Sample characteristics Residual chlorine

%RPD = {(S -D) /[(S+D)/2]) x 100" number (mg/L)

(acceptable range = k 10%)

1,70916 5

1 03"'llIftt S (-6.0017-Duplicate I

-. oC1 S Sample measurements:

Sample Sample ID Sample characteristics Residual chlorine number Reagent Blank r 0.00?6 k

eLsg..__.___---_

v2A&tlS.. to c..6SQWti.

no\\

CcAr,clear lZ 08.1~

C.. A-CO).(~

co (4 r c(er, pqr4Les L.O.00 t53 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 1

number (nmg/L)

(11g/L)

(acceptable range = 90 to 110%) I Z.4 1 C.1 0.50

o. cs q*.o%

Reviewed by Date reviewed SOP C8 - Exhibit C8. 1, revision 06-01-11 Page 71 of 99

Page 7

Page

_ of I "ETS Env*ronmentalTestlng 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 a's Date analyzed o*e.1.lZ Calibration:

[

Iodide reagent: [I t s7i7c Acid reagent:- [-*ItI-

-0.10 mg/L 1

1.00 mR/L 7

Slope Reference standard number I Irtss 10C4 1

r.qac.

Note: For samples with a residual chlorine of > 1.0 mg/L, the samples must be diluted to be within' the calibration range.

Laboratory control standard:

Reference standard True value (TV)

Measured value (MV)

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

(mg/L)

(acceptable range = 90 to 110%

J Djss 1o0q 0.50 o.SZo i0'.o%

J Duplicate sample precision:

Sample Sample ID Sample characteristics Residual chlorine

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

Duplicate D

0

'.001 co

.tM Sample measurements:

Sample Sample ID Sample characteristics Residual chlorine number

,__m__/L_

Reagent Blank

" 0o. oo07 1 M000____

fl0

-5 A~

aaTAAbcolor, clce'r 12,086~

18 0 h

wo%

lh Poils: -Se1au, Jee.r-f Lo.06 4Lf j9J%96q 6

¶'IqA SQ~va 1

tot.003fy

'L, rtAce flcotorcd,,efe.

jPPAJ&S L~c3 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)

IMeasured value (MV)

%RS=MVITVx 100 number I(mgIL)

I(Mg/L)

(acceptable range = 90 to 110%)

16C4 i

0.50 Oaiqcit.I I

Reviewed by Date reviewed LJA0-LLJ SOP CS - Exhibit C8. 1, revision 06-01-11 1Page 72 of 99

Sequoyah Nuclear Plant Biomonitoring August 14 - 21, 2012 Appendix D Reference Toxicant Test and Control Chart Page 73 of 99

E SEnvironmental Testing Solutions, Inc.

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

1.0 I

I I

I

~

I I

I USEPA Control Limits (+ 2 Standard Deviations)

I I

1 7--

0.9 F-0.8 0.7 0.6 k....................................'

0.5 [-

I II I

I I

1.4 1.2 1.0

.J S0.8

>1 0.6

'ý 0.4 USEPA Warning and Control Limits (75 and 90 Percentile CVs) 1.4 1.2 I

I I

I

. I I

I I

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

I I

I 1.0 0.8 0.6 0.4 Test date

-.------ 7-day IC., = 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 IC25 +/- SA.10 or 5A75)

Control Limits (mean IC25 +/- 5 A.25, SA.90' or 2 Standard Deviations)

Graphs generated from associated excel spreadsheet.

Excel spreadsheet entered by: J. Sumner Reviewed b.

k

ýý age 74 of 99

0 Environmental Testing Solutions, Inc.

Test number

. Test date 7-day IC T

cT L Kga)

(g/L Kd)

Pimephales promelas Chronic Reference Toxicant Control Chart State and USEPA Laboratory Control Umits Sw Warning ULmits S3.s CT-2S CT+2S CT-SAs CTa+SA1 Laboratory Control ULmits SK"S CT - S.

CT + SA..

USEPA Warning Umits S,..

CT - S*.

CT + S&s USEPA Control Umits CT-S&9 CT+ Sa.g CV 1

2 3

4 5

6 7

8 9

10 11 12 13 14 15 16 17 18 19 20 08-09-11 0.67 09-13-11 0.66 10-04-11 0.73 10-11-11 0.76 10-18-11 0.67 11-08-11 0.78 12-08-11 0.77 02-07-12 0.77 02-14-12 o.74 03-06-12 0.64 03-06-12 0.68 04-12-12 0.71 05-08-12 0.77 05-15-12 0.71 06-05-12 0.72 06-12-12 0.77 07-10-12 0.76 07-17-12 0.73 08-07-12 0.70 08-14-12 0.81 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.72 0.71 0.71 0.71 0.71 0.72 0.72 0.73 0.05 0.62 0.80 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.80 0.08 0.62 0.79 0.15 0.56.

0.86 0.27 0.44 0.05 0.61 0.81 0.09 0.62 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.62 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.44 0.05 0.62 0.81 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0.44 0.05 0.62 0.81 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0AS 0.05 0.63 0.82 0.09 0.64 0.81 0.15 0.57 0.88 0.28 0.45 0.98 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.99 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.99 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.99 0.32 0.39 0.99 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.98 0.32 0.39 0.99 0.32 0.39 0.99 0.32 0.40 1.00 0.33 0.40 1.03 0.07 1.03 0.07 1.03 0.07 1.03 0.07 1.03 0.07 1.03 0.07 1.03 0.07 1.03 0.07 1.04 0.07 1.03 0.07 1.03 0.07 1.03 0.07 1.04 0.08 1.04 0.08 1.03 0.07 1.03 0.07 1.03 0.07 1.04 0.07 1.04 0.07 1.05 0.06 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 IC2s).

S = Standard deviation of the IC25values.

Laboratory Control and Warning Umits Laboratory control and warning limits were established using the standard deviation of the lCm3values 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.sA = Standard deviation corresponding to the 10"' percentile CV. (SAM = 0.12) 5a.us = Standard deviation corresponding to the 2 5P percentile CV. (S&2 = 0.21)

USEPA Control and Waming Umlts SA.7 = Standard deviation corresponding to the 7 5t percentile CV. (S&., = 0.38)

Sa~ss= Standard deviation corresponding to the 9 0" percentile CV. (SAga = 0.45)

CV= Coefficient of variation of the IC2s values.

USEPA. 2000. Understanding and Aceunting for Method Variability in Whole Effluent Toicty Applications Under the National Pollutant 0Dscharge Elimination Program. EPA-833-8-09-003.

US Envlronmental Protection Agency, Cininnati, OH.

File: ppktci_081412Jdsxt Entered by:. J. Sumner Reviewed by. 99-

0 0

Environmental Testing Solutions, Inc.

I ".

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

-- 1.00 l2

0.75

-0.50 0 t0 E0.25 I

I I

I C

(

0 m

I s

USEPA Acceptance Criteria (> 0.25 mg per surviving larvae)

I I I

I I

.- I I

I I

F --

30 C

Kentucky Acceptance Limit (< 30.0%)

20

' a.

20 30 20 W. a-10 USEPA Upper PMSD Bound (90'h percentile < 30.0%)

012 0..

10 I

I I

III I

I I

I_

Test.d.......ate 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)

ýýage 76 of 99 Graphs generated from assodate1 excel spreadsheet.

Excel spreadsheet entered by: J :umner Reviewed by:

Precision of Endpoint Measurements SPimephales promelas EnvironmentalTestinggSolutions,.In.

Chronic Reference Toxicant Data Test number Test date Control Control Mean cv Cr MSD PMSD CT Survival Growth for Control Growth for Control W

for PMSD K

(mg/larvae)

N Growth CV (%)

1 08-09-11 100 0.662 6.6 0.07 10.6 2

09-13-11 100 0.848 0.755 6.7 6.7 0.12 13.6 12.1 3

10-04-11 100 0.768 0.759 16.8 10.0 0.13 16.9 13.7 4

10-11-11 i00 0.681 0.740 4.3 8.6 0.07 9.9 12.8 5

10-18-11 100 0.693 0.730 4.8 7.9 0.07 10.0 12.2 6

11-08-11 100 0.792 0.741 10.3 8.3 0.12 15.6 12.8 7

12-06-11 100 0.738 0.740 15.7 9.3 0.12 15.9 13.2 8

02-07-12 100 0.662 0.730 10.5 9.5 0.09 13.4 13.2 9

02-14-12 97.5 0.658 0.722 5.2 9.0 0.07 10.1 1.2.9 10 03-06-12 100 0.854 0.735 3.5 8.4 0.09 10.8 12.7 11 03-06-12 100 0.873 0.748 5.1 8.1 0.10 11.4 12.6 12 04-12-12 97.5 0.706 0.744 7.0 8.0 0.06 8.7 12.2 13 05-08-12 100 0.613 0.734 8.1 8.1 0.08 13.1 12.3 14 05-15-12 100 0.697 0.732 5.9 7.9 0.07 9.5 12.1 15 06-05-12 97.5 0.596 0.723 3.8 7.6 0.06 9.4 11.9 16 06-12-12 100 0.723 0.723 6.0 7.5 0.11 15.6 12.2 17 07-10-12 100 0.641 0.718 5.2 7.4 0.10 14.9 12.3 18 07-17-12 100 0.638 0.713 10.9 7.6 0.07 11.4 12.3 19 08-07-12 97.5 0.660 0.711 6.7 7.5 0.06 8.7 12.1 20 08-14-12 100 0.533 0.702 5.4 7.4 0.07 13.1 12.1 Note:

CV = Coefficient of variation for control growth.

Lower CV bound determined by USEPA (10d' percentile) = 3.5%.

Upper CV bound determined by USEPA (90td 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 (10th percentile) = 12%.

Upper PMSD bound determined by USEPA (901P percentile) =30%.

Cr = 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-O03. 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-821-8-01-005. US Environmental Protection Agency, Cincinnati, OH.

File: ppkclcr_0814]2.xlsx Entered by: J. Sumner age 77 of 99 Reviewed by: __

ETS Env~roarnentalTn~tngSouinIs Page I of 5 Potassium Chloride Chronic Reference Toxicant Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephales promelas PpKCICR Test Number: _1.

Dilution preparation information:

Comments:

KC1 Stock INSS number:

114sý

_o__

Stock preparation:

50 g KCI/L:

Dissolve 50 g KCI in 1-L MilHi-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:

__".oAX_____

0._3t)

Randomizing template:

Date and times organisms oa-b. r oiv. o Incubator number and were born between:

shelf location:

Organism source:

t Artemia CHM number:

CJltt L.

Drying information for weight determination:

Transfer vessel pH =

"1 eO*

S.U.

Date / Time in oven:

e6&t-

-a-i't information:

Temperature =

0,A 0C Initial oven temperature:

It Average transfer volume:

Date / Time out of oven:

fL.fl.

la

0.

'L "A

Final oven temperature:

1,

Total drying time:

mg-A*

Daily feeding and renewal information:

Day Date Morning feeding Afternoon feeding Test initiation, renewal, MHSW or termination batch used Time Analyst Time Analyst Time Analyst 0

__-_L

%So*0.

VL.S 090.0-02

,-IS.. I.

IMlt 0(3 1*t A(

It'~

ms

.) *t-IL.

4 at-I fl. I?..

01A1 0116 Mh~.

5__ 08 -Jr.r lgQ-I-61c AA~

4 j...

loPat-6 0110-mi..

O100 Vs oa 117. r%

at-16 I-).

-724 iot,

-21 Control information:

Acceptance criteria Summary of test endpoints:

% Mortality:

IT.

< 20%

.7-day LCs

&U. 'L(,

Average weight per initial larvae:.

O.-31 N

NOEC 00 Average weight per surviving larvae:

(.

1 2

! 0.25 mg/larvae LOEC

":! 0 ChV (016. no IC25 I

d1S (Iage 78 of 99 SOP AT21 -Exhibit AT21.1, revision 06-01-11

.ETS (b

Envtmnn-r0ITestlng Solutlom In Page 2 of 5 Species: Pimephales promelas PpKC1CR Test Number:

W)$_

Survival and Growth Data Day Control 450 mg KC[/L 600 m KCI/L

-.A B

C D.

E F

G H

I J

K L

0 101 0 1to It O

tO to 0 to I

/(

O /1

/0 1

10 10 to 1 10to 10 t

10 1.0

£

0)

/0 2

10 tO tO 1O to /o to

/O /O to /0 (O

3 (o

t (

(t 0

10 to t to 10 10 10 10 (0 4__

_1 10 10 10 10

l.

tO 0

qI&

1'A (0

/0 5

/0 /0

/0 t0 t0 /0 /0

/0 C

)

O 0

6

/

i U

10 "0

I1 I

/0 1

)

to(0 IO 7

tO_/0

_0 10

'O 1O0 to 10 'o'i to /0 A = Pan weight (mg)

Tray color code::

91;t, Analyst:.

J*

130*.*

4 i.q 1 48 '. 1 3.3 138 143 14.4 l t12. 1f.91 I

a.&Ij I3 ;

-&.4-2 Date:

0R.o1.t_

B = Pan + Larvae weight (mg)

Analyst:

40 1017 19 53 7A46 Iq.-3q 14.66 I'l.

Zo.ll V-50 19.50

.o.31 14.40 I9.1Z Date:

o 194.7..

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

S

,4

5.

s.

ie

.1 I.,.E1 AA*

q.L 6i.'* S.S S

(j.16 Analyst:

Weight per Initial number of larvae (mg)

= C /Initial number of larvae Hand calculated.

Analyst:

0*

1/2 A,

'V

'4 0*

A,

'4 C,

0 1/2 0

0 IV Average weight per initial Percent number of larvae (mag) reduction.

from control 0.%

MI_

L.7 0

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

Comments.

I age 79 of 99 SOP AT21 - Exhibit AT21. 1, revision 06-0 1-11

SS*ETS Page 3 of 5 Species: Pimephales promelas PpKCICR Test Number-. -IS Survival and Growth Data Day 750 mi KCIL 900 mg KCI/L 1050 mg KCI/L M

N 0

P Q

R S

T U

V NV X

0

/D t0o 0

10 10 to I0 /0

/0 /0 /o

/0 4_b

_ t' i,

_"z 2_

20 M :~ to 0

1-

-Qa.~'

3 1C0 0%

Ilk l_.

cj** -

u

-i 5-I',x 6

A = Pan weight (mg)

Tray color code::

Analyst:

13s.

[4B17.

1 3.18 1zS 4.-.s*.

13.42 IZ.

U

14. it1.05 13 IS.

13.O&

13.1f Date: __

B = Pan + Larvae weight (mg)

Analyst:

,,.gj q.'a..

l rTBq f

17.

ZS I

i7.1O tq.7*

Date:

OfE't,.z C = Larvae weight (mg) = B - A Hand calculated.

qLii 4),s S.11. 3.1t. S,'S t*,'s aoA O.*S IlS&

Analyst:

Weight per initial number of larvae (mg)

= C / Initial number of larvae Hand calculated.

4.

.O 0

Analyst:...

A I'

Average weight per Initial Percent number of larvae (mg) reduction

.O.U from control io.

Comment codes: c = clear, d = dead,.fg = fungus, k = killed, m = missing, sk = sick, sm - unusually small, 1g = unusually large, d&r = decanted and returned, w = wounded.

V Comments:

ge 80 of 99 SOP AT'21 - Exhibit AT21.1, revision 06-01-11

(D 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 numben 255 Test dates:

August 14-21, 2012 cowetrbniIneM/LXM Itp!na MW hnumbrnrerof Reinrmehnof A.P-Wft (nfq 0 Pes-tan Lan-e.

rnia5j:

Wdlit/Sul*veitn MAffi-W61ht/Sunung. Ca.errM ofnmtirton iuh1tlInffm.nunrbnrof M..rurcta(ll )MuNJ ihti~klfth Co.:Mfetnfvnitticn Per-tredunaonofm I-banta nelet ns A-0

'-abWenfhme.

lea) urenrhmolme. ee mtv ane.lief number ofiur.

)

mnb)l (%j

-.- 4 m) (%)

(-1d A

10 10 13.63 19.27 5.64 0.564 0.564 Control B

10 10 13.46 18.53 5.07 0.507 0.533 5A 0.507 100.0 0.533 5A Not applicable C

10 10 14.89 20.40 5.51 0.551 0.551 D

10 10 14.13 19.24 5.11 0.511 0.511 E

10 10 13.88 19.66 5.78

. 0.578 0.578 450 F

10 10 13.43 19.19 5.76 0.576 0.607 5.9 0.576 100.0 0.607 5.9

-13.9 G

10 10 14.54 20.81 6.27 0.627 0.627 H

10 10 15.02 21.50 6.48 0.648 0.648

.1 10 9

13.81 18.50 4.69 0.521 0.469 600 J

10 9

14.69 20.32 5.63 0.626 0.586 8.2 0.563 95.0 0.558 11A K

10 10 13.62 19.40 5.78 0.578 0.578 94.5 L

10 10 1342 19.62 6.20 0.620 0.620 M

10 8

13.12 17.89 4.77 0.596 0.477 750 N

10 9

14.56 19.24 4.68 0.520 0.547

7.

0.468 87.5 0.478 5.2 10.5 O

10 9

13.78 18.31 4.53 0.503 0.453 P

10 9

12.86 17.98 5.12 03569 0.512 Q

10 5

14.59 17.69 3.10 0.620 0310 900 R

10 6

13.72 17.28 3.56 0-593 0.607 4.4 0.356 57.5 0.348 12.4 34.8 5

10 7

12.18 16.23 4.05 0.579 0.405 T

10 5

14.11 17.30 3.19 0.638 0.319 U

10 1

12.85 13.36 0.51 0.510 0.051 1oso V

10 2

13.13 14.76 1.58 0.790 0.650 30.5 0.158 7.5 0.052 142.6 90.2 W

10 0

0.00 0.00 0.00 0.000 0.000 X

10 0

0.00 0.00.

0.00 0.000 0.000 Dunnett's MSD value:

PMSD:

0.0699 13.1 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 toxcity test.

Lower PMSD 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 Interlaboretory Variability Study (USEPA, 2001a; USEPA, 2001b).

USEPA. 2001a, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent ToxicityTest Methods, Volumes land 2-Appendix. EPA-821-1-01-004 and EPA-821-8-01-005. US Environmental Protection Agency, Cincinnati, OH.

Filme ppkcicr -_081412.xlsx Entered by: J. Sumner Reviewed b. _4

ET Statistical Analyses Larval Fish Start Date:

8/14/2012 Test ID:

PpKCICR End Date:

8/2112012 Lab ID:

ETS-Envii Sample Date:

Protocol: FWCHR-l Comments:

Conc-mglL 1

2 3

4 D-Control 1.0000 1.0000 1.0000 1.0000 450 1.0000 1.0000 1.0000 1.0000 800 0.9000 0.9000 1.0000 1.0000 750 0.8000 0.9000 0.9000 0.9000 900 0.5000 0.8000 0.7000 0.5000 1050 0.1000 0.2000 0.0000 0.0000 3rowth and Survival Test-7 Day Survival Sample ID:

REF-Ref Toxicant

r. Testing Sol.

Sample Type:

KCL-Potassium chloride EPA-821-R-02-013 Test Species:

PP-Pimephales promelas Transform: Arcsin Square Root Conc-mg/L Mean N-Mean Mean Min Max CV%

N D-Control 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4

450 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4

600 0.9500 0.9500 1.3305 1.2490 1.4120 7.072 4

750 0.8750 0.8750 1.2138 1.1071 1.2490 5.846 4

'900 0.5750 0.5750 0.8820 0.7854 0.9912 11.405 4

1050 0.0750 0.0750 0.2757 0.1588 0.4636 53.294 4

Rank 1-Tailed Sum Critical 18.00 10.00 14.00 10.00 10.00 10.00 10.00 10.00 10.00 10.00 Number Total Resp Number 0

40 0

40 2

40 5

40 17 40 37 40 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.93302 0.884 0.39148 0.28113 Equality of variance cannot be confirmed Hypothesis Test (1-tall, 0.05)

NOEC LOEC ChV TU Steel's Many-One Rank Test 600 750 670.82 Treatments vs D-Control Trimmed Spearman-Karber Trim Level EC50 95% CL 0.0%

5.0%

10.0%

902.19 867.55 938.21 1.0 20.0%

909.49 869.16 951.70 Auto-7.5%

898.26 863.27 934.67 0.9 0.8 0.7 0.6 0 0.5 0.4 0.3-0.2 0.1, 0.0, 1

10 100 1000 10000 Dose mgIL Dose-Response Plot 1 1 0.9 0.8 0.7 0.6 030.5 S0,4 0.3 0.2 0.1

age 82 of 99 File: ppkclcr.0814l:t.xlsx Entered by:.. Sumner Reviewed by:

ETS 0 nl~~?a ~~g.oulkI(

Statistical Analyses Start Date:

8/1412012 End Date:

8/21/2012 Sample Date:

Comments:

Conc-mglL 1

2 D-Control 0.5640 0.5070 450 0.5780 0.5760 600 0.4690 0.5630 750 0.4770 0.4580 900 0.3100 0.3560 1050 0.0510 0.1580 Larval Fish Growth and Survival Test-7 Day Growth Test ID: PpKCICR Sample ID:

REF-Ref Toxicant Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

KCL-Potasslum chloride Protocol: FWCHR-EPA-821-R-02-013 Test Species:

PP-Pimephales promelas 3

4 0.5510 0.5110 0.6270 0.6480 0.5780 0.6200 0.4530 0.5120 0.4050 0.3190 0.0000 0.0000 Transform: Untransformed Conc-mg/L Mean N-Mean Mean Min Max CV%

N D-Control 0.5333 1.0000 0.5333 0.5070 0.5640 5.353 4

450 0.6073 1.1388 0.6073 0.5760 0.6480 5.924 4

600 0.5575 1.0455 0.5575 0.4890 0.6200 11.433 4

750 0.4775 0.8955 0.4775 0.4530 0.5120 5.244 4

900 0.3475 0.6517 0.3475 0.3100 0.4050 12.430 4

1050 0.0523 0.0980 0.0523 0.0000 0.1580 142.558 4

1-Tailed t-Stat Critical

-2.307 2.180

-0.756 2.180 MSD 0.0699 0.0699 Isotonic Mean N-Mean 0.5703 1.0000 0.5703 1.0000 0.5575 0.9776 0.4775 0.8374 0.3475 0.6094 0.0523 0.0916 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test Indicates normal distribution (p > 0.01) 0.94803 0.805

-0.6553 0.58729 Bartlett's Test Indicates equal variances (p = 0.40) 1.85289 9.21035 Hypothesis Test (1-tall, 0.05)

NOEC LOEC ChV TU MSDu MSDp MSB MSE F.Prob df Dunnett's Test 600

>S00 0.06992 0.13112 0.00569 0.00206 0.11571 2,9 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point mg/L SD 95% CL(Exp)

Skew IC05 629.55 44.05 422.75 677.99

-1.2511 IC10 683.02 39.17 471.16 745.11

-1.6458 IC15 736.48 32.27 546.98 788.00 -2.5024 1.0 IC20 774.58 14.39 725.29 814.40 0.0156 08 IC25 807.48 14.81 783.34 852.44 0.1502 0.8 IC00 902.72 14.24 850.75 928.37

-0.5101 IC50 931 6i9 908R 905.24 9-5694 01-578 0.7-0.6 0.5 CO 0.4 I0.3 0.2~

500 1000 1500 Dose mg/L Dose-Response Plot 1.04 0.3 0.2-1-tall, 0.05 level of significance (V

age 83 of99 Vile: pplciattl1l2.xIs Entered by: Jl. Suniner Remiwed byr.-

00ITSý Envlmonment.Tesfn Sltl o I Page 4 of 5 Species: Pimephales promelas PpKC1CR Test Number: _IS Daily Chemistry:

Day Aaddtified for each da erformed D.O. and conductivity measurements onlyv.

Concentration Parameter pH (S.U.5) 77.*

Ss.*.

D O (m g/L)".

7...

Conductivity 0

o CONTROL

Alkalinity (mg CaCO3/L)

O-

" *Hardness (mg CaCO3IL) i '1

,____*Temperature (OCD.

VA.

A.

. it pH (S.U.)

.7.73".

DO (mg/L) 7.7 77 450 mg KCI/L Conductivity

Temperature (IC)

"k.4. -

i.

S pH (S.U.)

1.*S i.s7 l.o 7-4SZ.

7oI DO (mg/L) 14

7.

7.8 600 mg KCIIL Conductivity mhos/cm)

(C 133

%360 bV N

Temperature (0QD -

C

7.

-3

4.

DO (mg/L)

"7.7 "7.0 1

750 mg KCIIL Conductivity (gmhos/cm) 100 1 CAO

Temperature (op

14. 46 T41 pH (S.U.)

a

.5?-4

.C DO (mg/L) 7.9 900 Ing KCIIL LConductivity

(~mhoslcm)

Is1&

Temperature (0) l'4q

-Lq.14.

2.

ii

.ýq pH (S.U.)

'711 II7c:;

1 -7.-q I ~113 1 : -18 11 14.--

1 DO (mg/L) 1050 mg KCI/L Conductivity (jLmh sc)

-T Z O70

Temperature (IC)

STOCK Conductivity

-'qo0 (Rmho Initial II Final I Initial II Final I Initial 11 Final I\\

Ix

Temperatures perfobmed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page Alkalinity and hardness performed by the analyst. identified on the bench sheet specific for each analysis and transcribed to this bench sheet by:

age 84 of 99 SOP AT21 -Exhibit AT21.1, revision 06-01-11

OET E~wlmmeul'uTesung

&olIw~lo.In Page 5 of 5 Species: Pimephales promelas PpKC1CR Test Number:

21K Day (Analyst identified for each day, performed pH, D.O. and conductivity measuriements only.)

3 4

5

_____Analyst am zru3 S

0

sf V

Concentration Paramter pH (S.U.)

7.CO 7.,7 V...

".I DO(mg/L) 7..

i i.Ci Conductivity (jinhoscm)31 CONTROL

Alkalinity (nig CaCO3/L) 12

Hardness ing CaCO3/L)

Temperature pH (S.U.)

.7.7.I 7.4 71o 7,L..

7.l

.S DO (mg/L) 450 mng KCI/L Conductivity (l'imhos/cm)

O)

Temperature pH(S.U.)

O3.-+

-7.S7 7.70

-.7..77

-i.

itS 7.7.

1 DO (mgiL) 7.

50 nmg KCIL Conductivity

-(gmhos/m) 1

Temperature U

pH (S.U.)

-3 Q

y.

7.77

.73:77 1i I

_._7'.

DO (mg/L) 7.__

.7.'7 900 mg KCI/L Conductivity hTempe r

.,6

Temperature i*.o

-iM.

2.M.1 *L*.I. 0

-Ll.6.

-A,.

i,.44

-uA.S pH (S.U.)

-a, 7.C-'

-i.'iq 7

Lj 7.1

-7.-

7.7q 7.$

DO (mg/L)

-7 17.3 7,C

7.

goo0mg KCIIL Conductivity41tO

Temperaturei"!

S A..

L.l. ','

"1.

(Oc) 0M

-6 4

Initial I Final. I Initial I Final I Initial 11 Final I Initial I Final I

U U

0, i

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located qa Page 1.

Alkalinity and hardfiess performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by: 7 V

(f age 85 of 99 SOP AT21 - Exhibit AT21.1, revision 06-01-11

Ec Environmental Testing Solutions, Inc.

Ceriodaphnia dubia Chronic Reference Toxicant Control Chart 1.14 1.12 1.10 -

1.08 1.06 1.04 1.02 2.5 I

I I

l I

I I

USEPA Control Limits (k 2 Standard Deviations)

I I

t II I

I I

zJ:

N, 2.0 1.5 1.0 0.5 1.4 1.3 I

I I

I Laboratory Warning and Control Limits (10th and 2 5 th Percentile CVs) 1.2 H-1.1 1.0 -

0....

7-0.9 0.8 I

II I

Ifi fI~

I I~ L I

I INN NN NN NN NN -IN NN INN N'111 NN N$ N$ 0 N$ -'s N, Ns

-o WNs Ok"',

NO-N" OSA Test date 7-day IC25 = 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 IC2s)

Warning Limits (mean IC 2 5 1 SA1 0 or SA.75)

Control Limits (mean IC25 + SA 25, SA90' or 2 Standard Deviations) age 86 of 99 Graphs generated fromassociated excel spreadshleet.

Excel spreadsheet entered by: J. Sumner Reviewedby fY

.ETS 8 Environmental Testing Solutions, Inc.

Ceriodaphnia dubia Chronic Reference Toxicant Control Chart Test number Test date 7<day IC2, CT (g/L NaCI)

(WJL NaCI) 1 2

3 4

5 6

7 8

9 10 11 12 13 14 15 16 17 18 19 20 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 12-06-11 01-10-12 02-07-12 03-13-12 04-10-12 04-10-12 05-08-12 05-08-12 06-05-12 07-10-12 08-07-12 1.07 1.07 1.05 1.06 1.06 1.06 1.07 1.06 1.05 1.06 1.08 1.08 LOS 1.07 1.07 1.06 1.05 1.05 1.07 1.08 1.07 1.07 1.07 1.07 L07 1.07 1.07 1.07 1.07 1.06 1.07 1.07 1.07 1.06 1.06 1.06 1.06 1.06 1.06 1.06 State and USEPA S

Control Limits SAM CT-2S CT+2S 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 1.04 1.09 0.09 0.01 1.04 1.09 0.09 0.01 1.04 1.09 0.09 0.01 1.05 1.08 0.09 0.01 1.05 1.08 0.09 0.01 1.04 1.08 0.09 0.01 1.04 1.09 0.09 0.01 1.04 1.09 0.09 0.01 1.04 1.09 0.09 0.01 1.05 1.08 0.09 0.01 1.05 1.08 0.09 0.01 1.05 1.06 0.09 0.01 1.05 1.08 0.09 0.01 1.05 1.08 0.09 0.0.1 1.05 1.08 0.09 0.01 105 1.08 0.09 Laboratory Warning Umits CT - S&.

CT

SX0 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0-98 0.98 0.98 0.98 0.98 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 1.15 1.15 1.15 1.15 1.15 215 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 0.18 Laboratory SA25 Control Limits CT-SAý CT + Sz USEPA SAL3 Warning Limits CT-SA-s CT+ SA.7 USEPA S,.O Control Limits CT - S,,

CT+Sjue CV 0.89 0.89 0.89 0.89 0.89 0.89 0.89 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 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 1.25 1.24 1.24 1.24 1.24 1.24 1.25 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 0.48 O.48 0.48 0.48 0.48 0.48 0.48 0A8 0A8 0.48 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 0.59 O.59 0.59 0.59 0.59 0.59 0.59 0.58 0.58 0.59 0.59 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.55 1.54 1.54 1.54 1.55 1.55 1.54 1.54 1.54 1.54 1.54 1.54 1.54 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 0.66 0.66 0.66 0.66 0.66 0.66 0.66 0.41 0.41 OA1 0.41 0.41 0.41 0.41 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 1.73 1.73 1.73 1.73 1.73 1.73 1.73 1.73 1.73 1.72 1.73 1.73 1.73 1.72 1.72 1.72 1.72 1.72 1.72 1.72 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 0.01 Note:

7-d c1, = 7-day 25% inhibition concentration. An estimation of the concentration of sodium chloride that would cause a 25% reduction in Ceriodaphnlo reproduction for the test population.

CT = Central tendency (mean IC-s).

S = Standard deviation of the ICn values.

Laboratory Control and Warning Umits Laboratory control and warning limits were established using the standard deviation of the ICE 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 Sý. = Standard deviation corresponding to the 10e percentile CV. (SAe. = 0.08)

$*j, = Standard deviation corresponding to the 2P percentile CV. (SA25 = 0.17)

IJSEPA Control and Warning Umits SAJ. = Standard deviation corresponding to the 75h percentile CV. (S&75 = 0.45)

SeA"= Standard deviation corresponding to the 9 0e percentile CV. (S,

= 0.62)

CV = Coefficient of variation of the IC, values.

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

File: CdNca0CCRO8Ol72icdsx Entered by: J. Sumnjr Reviwemd by-.:

ET:

Environmental Testing Solutions, Inc.

Ceriodaphnia dubia Chronic Reference Toxicant Control Chart Precision of Endpoint Measurements 40 00 0I.'

0 00

U M

35 30 25 20 15 I

I I

J I

I I

i I

I Minimum Acceptance Criteria (> 15.0 offspring per surviving female)

I I I

I I

I

o40

.0 O 30 o-20

~10 50 North Carolina Acceptance Limit (< 40.0%)

Kentucky Acceptance Limit (< 30.0%)

i!!...................-..........................................

40 30 20 10 11

..I I

I I

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

S...................

I I

V Ns 'N'

,'SAI, 'bx'ý I I.

O'vs sqýv

, 'N SN NN

-N'* 1'ý-

NAN N-N"' %A-" 0'*

OINI -

o1b bg-" "b"i "'b" ANý%

NTI

'bT lo, lo,

'o-1 Q'S" 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)

I 4-

ý f -nlet

ý age 88 of 99 Graphs generated from associate I excel spreadsheot Excel spreadsheet entered by:..;umner Reviewed by:

lPrecision of Endpoint Measurements

.E T S 1Ceriodaphnia dubia IChronic Reference Toxicant Data Environmental Testing Solutions, Inc.

Test Control Control Mean Test date CT CV CF MSD PMSD

(.Ti number Survival Reproduction for Control Mean for Control (N)

(offspring/female)

Reproduction (K)

Reproduction

(%)

for PrVlSD (%)

(offspring/female)

CV (%)

1 04-05-11 100 33.0 5.9 2.2 6.7 2

05-03-11 100 30.9 32.0 4.5 5.2 1.9 6.1 6,4 3

06-07-11 100 28.9 30.9 6.7 5.7 2.5 8.7 7.2 4

07-12-11 100 32.2 31.3 6.0 5.8 2.0 6.2 6.9 5

08-09-11 100 28.3 30.7 5.0 5.6 1.9 6.8 6.9

.6 09-13-11 100 32.1 30.9 8.2 6.1 1.9 5.8 6.7 7

10-04-11 100 32.8 31.2 6.1 6.1 2.1 6.5 6.7 8

10-11-11 100 31.3 31.2 3.0 5.7 2.1 6.6 6.7 9

11-08-11 100 29.0 30.9 7.6 5.9 1.8 6.1 6.6 10 12-06-11 100 29.3 30.8 5.8 5.9 1.9 6.5 6.6 11 01-10-12 100 32.4 30.9 5.1 5.8 2.4 7.4 6.7 12 02-07-12 100 31.7 31.0 4.2 5.7 2.1 6.5 6.7 13 03-13-12 100 31.1 31.0 7.2 5.8 1.9 6.1 6.65 14 04-10-12 100 29.4 30.9 6.8 5.9 2.4 8.0 6.7 15 04-10-12 100 30.3 30.8 7.1 6.0 2.7 8.8 6.13 16 05-08-12 100 31.7 30.9 6.1 6.0 1.9 6.1 6.13 17 05-08-12 100 32.4 31.0 5.1 5.9 2.0 6.1 6.3 18 06-05-12 100 32.1 31.1 4.0 5.8 2.3 7.1 6.8 19 07-10-12 100 31.2 31.1 4.7 5.8 2.0 6.3 6.18 20 08-07-12 100 29.2 31.0 3.5 5.6 1.9 6.6 6.7 Note:

CV = Coefficient of variation for control reproduction.

Lower CV bound determined by USEPA (10' percentile) = 8.9%.

Upper CV bound determined by USEPA (90'h 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 (10th percentile) = 13%.

Upper PMSD bound determined by USEPA (90th percentile) = 47%.

Cr = 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 Discharlge 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 l and 2-Appendix. EPA-821-B-01-004 and EPA-821-B-01-005. US Environmental Protection Agency, Cincinnati, OH.

File: CdNaCICRO080712.xlsx Entered by: J. Smnner (t

age 89 of 99 Reviewed by: _*-

0ETS Page I of 6

/'

i Sodium Chloride Chronic Reference Toxicant Test (EPA-821-R-02-013 Method 1002.0)

Species: Ceriodaphnia dubia CdNaCICR #:

I.*

Dilution preparation in ormation:

Comments:

NaCI Stock INSS number.

ltyA.

__n_

Stock preparation:

100 g NaCI/L:

Dissolve 50 g NaCI in 500 mL Milli-Q water.

Dilution prep (mg/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 1500 1500 1500 1500 Test organism source information:

Test information:

Organism age:

< 24-hours old Randomizing template color Date and times organisms were bom mGL.01t U. 01% M toW Incubator number and shelf between:

location:

Culture board:

Replicate number:

2 3 )4 15 16 17 a

9 10 YWT batch:

Culture board cup number-I 'I 1

oi 6% 110 1-010_________

Transfer vessel infbrmation:

pH=

i".61 S.U1 Temperature = 2q.t *C Selenastrum batch:

Average transfer volume (mL):

0.0 51

-A Daily renewal information:

Day Date Test initiation and feeding, MHSW Analyst renewal and feeding, or batch used termination time 0

ot.-.(t.

tOi-oI -OsrL 2

0.f_____-a

.___5 oiq1S 04-0-h11-~ 4..

6.

.O

-. 1"I.

CA *.\\,

t 0 Control information:

Acceptance criteria Summary of test endpoints:

% of Male Adults:

0

.20%

7-day LC50 51400

% Adults having 3rd Broods:

INA 80%

NOEC 000

% Mortality:

f7.

520%

LOEC I*

t3 Mean OffspringFemale:

21

> 15.0 offspring/female ChV 101K.

%CV:

_,S7.1

<40.0 %

IC 25 161 (f

age 90 of 99 SOP AT14 - Exhibit AT14.1, revision 06-01-11

S6ETS Page 2 of 6 Species: Ceriodaphnia dubia CONTROL

/

CdNaCICR #:

Vbt Survival and Reproduction Data II

- I....

Replicate number Day 1n 11 2

3 4

5 6

7 8

9 10 21 Young produced I....bi..l

. 1l o o

I 600 1.~.

Adult mortality I

%. I t-L L-i -

Youngprodueed n

in II n 0-Adult mortality [__

.-'%-I L- ~II~ t-- I %- 'I

% -- I FL I--

3 Young produced Adult mortalitg

.*q a

I Adult mortality Tot5l young produced "T..."1 Final Adult Mortality CT f '

i._.._.

C_

X for ung roods C

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

Cnetao:

OT A % Mortality:

Mean Offs rin eFmale:

[ ",1.Lt-600 NaCI/L Survival and Reproduction Data I

~"

Replicate number 4

%S I'S Day' 1d 2

3 4

- 6

[ 8 9

10 Toa Iyoung produced 3

v%

1w ft

__IL1

-LI Adult mortality 17

_N

_7 2

Y oungproduced jJ ol lo

)o.. *I() Q.

I o N Adult mortality I.LIt b-o (l.s I

brodI s.p.li b--1 t1i Young producedl 1

10 0

t r Ins0 I

r I_ Admlt mortality LFWT*.

JC]--

t I

[T7I I '-

I-1 I

,4 Young produced i~ S 113"1 I*"I*

1*I I '.

I Adult mortality 1

I I

'I%

Mot ality

[107.

Adultmortallty 'I__l.

JMe 1

.f.

iJ.

n 1+/-1

[

60 mVug NC/

uvvladRproducto Data:*

Iq Day 1

J 1__

314 1

8_

9 10 It Young produced j

oi 0

3 0

"5 1

20 l__ Adult mortality IA~

L I

L-I ['Lr L L L.._

L i-L 2 Young produced (n

4

-0II Toa Young produced 1.0~3

~~

b 2

Final Adult Mortality TJ -

t.

Note: Adult mortality (L = live, D L

dead), SB = split brood (single brod split between two days), CO = carry over (offspring carried over with a dluring ansfer).

Concentration:

% Mortality:

07.

Mean Offspring/Female:

0o.2-.

% Reduction from Control:

-.a 7.

SOP AT14 - Exhibit ATI4.1, revision 06 11

%p ge 91 of 99

6ETS Page 3 of 6 Species: Ceriodaphnia dubia 800 ma NaCI/L CdNaCICR #: I_&

Survival and Reproduction Data Replicate number Da y

1 2

3 4

5 6

7 8

9 10 I

Young produced 0

0..

1) 0 0C1 0

_ Adlmortality[

~

N.-

I~

2 Young produced Adult mortality a

s..

3 oung produced 0

)

0J O0 01 0

Adult mortality

- I I I ___________________________

4 Young produced

"%I t,

Adult mortality S

Young produced 103J

~I~Ii 17-1 06I__ 2.

C Adult mortality

.... *q-.

6 oung produced

__S_

Adult mortality I

I 7

Young produced Total young produced z

"%..w B0%

'Lt Final Adult Mortality X__

j_ _

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

Concentration:

% Mortality:

07.

Mean Offspring/Female:

.30A

% Reduction from Control:

k 17.

1000 mg NaCI/L Survival and Reproduction Data Replicate number Day 1

2 3

4 5

6 7

8 9

1 10 1

Young produced

]

I-i Adult mortality t,.-*_

I 2oa Young produced o*

n}

b 6

nalAdult mortality.L L

I U-.-

UI. L,-

3f Young produced C) I e)

Cp oe Adult mortality W

t L Iro I

LrIoL spItw w

rr i

a4 Young produced

JA s6 I I Adult mortality JL

%-~1

.~J

.J...

I 5

Young prdue 0C 1_ '1,t 10 IP I1.

[Adult mortality V_

.6 Young produced ti

0.

0 0

0 Adult mortality X4_

I__ LJ__%_I

'- I V-- I

-I 7 Young producedJ %S 'S

~

IS 16 17'.

IS, i-Final Adult Mortality 7-k L -'

Note: Adult fliortality (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:

10.q Mean Offspring/Female:

24.q age 92 of 99 I% Reduction from Control:

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

0ETS Page 4 of 6 Species: Ceriodaphnia dubia 1200 me NaCI/L CdNaCICR #: _

Survival and Reproduction Data Replicate number 1

Young produced Q

Qt III Adult mortality 1

.3 4

5 6

7 8

9 10 1

Young produced 1

Adult mortality A.--'-__

-P I'

1_-'-

3 Yugpoue0-n 0_l

_J Adult mortality I~I

_it C4CJ 4

Young produced I.*jZ*

1 2I

,o I.-

L j I I

__IAdult mortality U J J'

I'-

L 5

oung produced I

I I

_ Adult mortality. I 1'".-

['-

"FTIL-"

Adult mortality 7

Young produced 5

i --

I

-1

%A Total young produced jA I1S 12-66 1

16 Final Adult Mortality

-Q-

%?

N

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

Concentration:

% Mortality:

0?.

.Mean Offspring/Female:

%.L

% Reduction from Control:

SI'&.

1400 mg NaCI/L Survival and Reproduction Data Replicate number Day 1 1 2

3 4

5 6

7 8

9 10 1I Young produced Adult mortality I L

QL CL C

1 E. t 2

Yongproduced cc)

I rr

()

ia SAdult mortaliayt y*

_I--I*

I I I L _

3 Young producedI 0.

O I

I

'4 Young produced Adult mortality 3

Young produced Adult mortality I IU I.

LI_

u,._

1 L.

L.... i 6

Young produced 0J.JA I~

IL t

~

1.. 0.~

Adult mortalityj l

.4U L

IL L

U j

7 Young produced

. X 1

Total young produced Lk A

10S01 L j C)

Final Adult Mortality Note: Adult mortality (L = live, D =dead), SB = split brood (single brood split between two days), CO = carry over (offspring age 93 of 99 Concentration:

Mortality:

07.

Meann Offilspring/Female:

1,,.0 I% Reduction from Control:

71.

$1.4 SOP AT14 - Exhibit AT14. 1, revision 06-01-11

!E TS F nvlnrnmental Testing Solutions, Inc.

Verification of Ceriodaphnia Reproduction Totals Control Day Replicate number Total Dy 1

21 3 4

5 6

7 8

9 10 1

0 0

0 2

0 0

0 3

0 0

0 4

4 5

4 4

5 5

6 4

5 5

47 5

11 0

10 10 10 10 12 10 10 10 93 6

0 10 0

0 0

0 10 7

15 13 15 15 14 15 13 16 13 13 142 Total 30 28 29 29 29 30 31 30 28 28 292 600 mg NaCI/L Day Replicate number Total ay 1

2 3 -4 5

,6 7

8 9

1 oa 1

0 0

0 2

0 0

0 3

0 0

0 4

5 5

4 6

6 4

6 5

5 5

51 5

11 0

10 10 13 11 11 10 13 10 99 6

0 11 0

0 0

0 11 7

14 13 14 14 1

1 4

13 13 141 Total 30 29 28 30 33 31 33 29 31 28 302 800 iNg aCi/L Day Replicate number Total 1

2 3

4 5

6 7

8 9

10 1

0 0

0 2

0 0

0 3

0 0

0 4

5 5

4 4

4 5

4 6

6 5

48 5

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

0 0

00 0

7 14 14 14 13 13 16 15 17 13 14 143.

301 Total 30 30 28 28 27 33 29 36 31 29 301 1000 mg NaCI/L Replicate number Total Day 1

2.

3 4

6 7

8 19 1 T10 1

0 0

0 2

0 0

0 3

0 0

0 4

5 4

6 4

4 4

4 3

4 4

42 5

0 12 11 11 10 10 10 10 10 10 94 6

11 0

0 0

11 7

15 15 13 15 12 14 13 13 13 14 137 Total 31 31 30 30 26 28 27 26 27 28 284 1200mg NaCLa Day Re cate number Total I

1 2

3 4

5 6

7 8

9 10 1

0 0

0 2

0 0

0 0 10

.0 0

0 0

3 0

0 0

4 3

1 2

2 3

2 1

3 3

3 23 5

7 0

6 4

5 8

5 6

9 5

55 6

0 9

0 0

9 7

4 6

5 3

5 2

7 4

4 5

45 Total 14 16 13 9

13 12 13 13 16 13 132 1400mni NaCIIL 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

1 1

1 2

2 3

3 1

19 5

0 0

2 3

0 2

4 1

2 2

16 6

1 3

0 0

2 0

0 0

6 7

2 2

1 1

1 3

2 3

1 3

19 Total 4

6 4

6 5

8 8

7 6

6 60 File: CdNaCICR_080712.xlsx Entered by: J. Sumner Reviewed by:

CA 0

too Ceriodaphnia dubia Chronic Reference Toxicant Test EPA-821-R-02-013, Method 1002.0 is Environmental Testing Solutions, Inc.

Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Test number:

Test dates:

CdNaCICR #138 August 07-14,2012 Concentration Replicate number Survival Average reproduction Coeffident of Percent reduction from (mg/L NaO) 1 2

3 4

5 6

7 8

9 10

(%)

(offspringfemale) varIatton (%)

control (%)

Control 30 28 29 29 29 30 31 30 28 28 100 29.2 3.5 Not applicable 600 30 29 28 30 33 31 33 29 31 28 100 30.2 6.0

-3.4 800 30 30 28 28 27 33 29 36 31 29 100 30.1 8.9

-3.1 1000 31 31 30 30 26 28 27 26 27 28 100 28.4 6.9 2.7 1200 14 16 13 9

13 12 13 13 16 13 100 13.2 15.1 54.8 1400 4

6 4

6 5

8 8

7 6

6 100 6.0 23.6 79.5 Dunnett's MSD value:

PMSD:

1.929 6.6 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 PMSD bound determined by USEPA (I0 th percentile) = 13%.

Upper PMSD bound determined by USEPA (90'h 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. 2001a, 200lb. 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: CdNaCICR_080712.xlsx Table populated from associated 'Verification of Cedodaphnia Reproduction Totals" spreadsheet.

Spreadsheet entered by: J. Sumner Reviewed by:

1

SETS 6

nWrwrmWeazft oou~wwIm Statistical Analyses Ceriodaphnla Survival and Reproduction Test-Reproduction Start Date:

8/712012 Test ID:

CdNaCICR Sample ID:

REF-Ref Toxdcant End Date:

8/14/2012 Lab ID:

ETS-Envlr. Testing Sol.

Sample Type:

NACL-Sodium chloride Sample Date:

Protocol: FWCHR-EPA-821-R-02-013 Test Species:

CD-Cariodaphnia dubla Comments:

Conc-mgIL 1

2 3

4

.6 6

7 8

9 10 D-Control 30.000 28.000 29.000 29.000 29.000 30.000 31.000 30.000 28.000 28.000 600 30.000 29.000 28.000 30.000 33.000 31.000 33.000 29.000 31.000 28.000 800 30.000 30.000 28.000 28.000 27.000 33.000 29.000 36.000 31.000 29.000 1000 31.000 31.000 30.000 30.000 26.000 28.000 27.000 26.000 27.000 28.000 1200 14.000 16.000 13.000 9.000 13.000 12.000 13.000 13.000 16.000 13.000 1400 4.000 6.000 4.000 6.000 5.000 8.000 8.000 7.000 6.000 6.000 Transform: Untransformed Conc.mg/L Mean N-Mean Mean Min Max CV%

N D-Control 29.200 1.0000 29.200 28.000 31.000 3.537 10 600 80.200 1.0342 30.200 28,000 33.000 6.005 10 800 30.100 1.0308 30.100 27.000 36.000 8.921 10 1000 28.400 0.9726 28.400 26.000 31.000 6.884 10

'1200 13.200 0.4521 13.200 9.000 16.000 15.067 10

'1400 6.000 0.2055 6.000 4.000 8.000 23.570 10 Rank 1-Tailed Sum Critical 121.00 75.00 111.50 75.00 92.00 75.00 55.00 75.00 55.00 75.00 Isotonic Mean N-Mean 29.833 1.0000 29.833 1.0000 29.833 1.0000 28.400 0.9520 13.200 0.4425 6.000 0.2011 Auxiliary Tests Statistic Critical Skew Kurt Kolmogorov D Test indicates non-normal distribution (p <= 0.01) 1.17686 1.035 0.53863 0.89662 Bartlett's Test indicates equal variances (p = 0.14) 8.40619 15.0863 Hypothesis Test (1-tail. 0.05)

NOEC LOEC ChV TU Steers Many-One Rank Test 1000 1200 1095.45 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point ma/L SD 95% CL Skew IC05 1000.77 44.7051 868.009 1015.87 -1.6465 IC10 1020.39 9.61319 991.422 1034.38

-1.3975 IC15 1040.02 7.68096 1020.26 1053.25

-0.3510 1.o-IC20 1059.65 7.19436 1041.68 1071.64 -0.3336 0.9 IC25 1079.28 6.83735 1063.08 1090.45

-0.3157 IC40 1138.16 6.71529 1123.99 1148.74

-0.2616 0.8 IC50 1177.41 7.42545 1162.25 1189.63 -0.2048 0.7 0.6 0.4 0.3 0.2 0.1.

0.0

-0.1 1 0

500 1000 1500 Dose mg/L Dose-Response Plot 30 25 20 10 5

(fage 96 of 99 File: CdNaCICR0807I2.xlsx Entered by: J. !;unmer Reviewed by. _-_

"ETS (a

Environmentl Tesoti ons, Inc.

Statistical Analyses Ceriodaphnia Survival and Reproduction Test-Reproduction Start Date:

8/7/2012 Test ID: CdNaCICR Sample ID:

REF-Ref Toxicant End Date:

8/14/2012 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:

Used for PMSD calculation only.

Conc-maiL 1

2 3

4 6

6 7

8 9

10 D-Control 30.000 28.000 29,000 29.000 29.000 30.000 31.000 30.000 28,000 28.000 600 30.000 29.000 28.000 30.000 33.000 31.000 33.000 29.000 31.000 28.000 800 30.000 30.000 28.000 28.000 27.000 33.000 29.000 36.000 31.000 29.000 1000 31.000 31.000 30.000 30.000 26.000 28.000 27.000 26.000 27.000 28.000 1200 14.000 16.000 13.000 9.000 13.000 12.000 13.000 13.000 16.000 13.000 1400 4.000 6.000 4.000 6.000 5.000 8.000 8.000 7.000 6.000 6.000 Transform: Untransformed 1-Tailed Conc-mg/L Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD D-Control 29.200 1.0000 29.200 28.000 31.000 3.537 10 600 30.200 1.0342 30.200 28.000 33.000 6.005 10

-1.186 2.287 1.929 800 30.100 1.0308 30.100 27.000 36.000 8.921 10

-1.067 2.287 1.929 1000 28.400 0.9726 28.400 26.000 31.000 6.884 10 0.948 2.287 1.929

1200 13.200 0.4521 13.200 9.000 16.000 15.067 10 18.969 2.287 1.929

1400 6.000 0.2055 6.000 4.000 8.000 23.570 10 27.505 2.287 1.929 AuxillaryTests Statistic Critical Skew Kurt Kolmogorov D Test indicates non-normal distribution (p <= 0.01) 1.17686 1.035 0.53863 0.89662 Bartletts Test indicates equal variances (p = 0.14) 8.40619 15.0863 Hypothesis Test (1 -tail, 0.05)

NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob CIf Dunnett's Test 1000 1200 1095.45 1.92879 0.06605 1109.51 3.55741 1.6E-38 5,54 Treatments vs D-Control

age 97 of 99 File: CdNaCICR_0807 [2.xlsx Entered by: J. Sumner Reviewed by: _A-

I.,.

.....:.e.5

,.:6 Page 5 of 6 Species: Ceriodaphnia dubia Daily Chemistrv:

CdNaCICR #:

IVb "Day (Aayst identified for each dayr performed pHl, D.O. and conductivity measure'ments only.

Concentration Parameter PH (s.u.)

[

Conductivity (gmihos/cm) 0 CONTROL

Alkalinity (Ing CaCO3/L)

Hardness (mg CaCO3 L)"

Temperature (*C).

A*

1¢4. 'i 1

pH (S.U.)

1.7C

.23 I DO (mg/L) 7.7 9" D 600 mg NaCIIL Conductivity (tuhhos/cm) 14t__'_13_

Temperature(*C)

-Lq A U_0 C) pH (S.U.)

M.U 147 DO (mgIL) 7.1 800 mg NaCI/L Conductivity

Temperature (9C) 1I4.0,

. 0

1. 0 all pH (S.'U.)

-7.77____

1000 mg NaCI/L Conductivity

Temperature (IC)

IS..0

,SO "S._,_

"-s.

pH (S.ff.)

"7,77/ f

.*4 I**Z 7.*7.S 0

8._

1200 mg NaCI/L Conductivity I *Temperature (OC) 75,.0

.1.,

I S.

pH (S.U.)

7.90 "4.91

~

I -

DO (mg/L) 1400 mg NaCIIL Conductivity o

(g+/-mhos/c)

Temperature (SC)

STOCK Conductivity t

1~

-gno/m Initial I1 1

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1. Alkalinity and hardness performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by:

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

-em.-,

b~,wby Kt.,EKnm e 98 of 99

S S0ETS E~maIT~OgSOI**b~

Page 6 of 6 Species: Ceriodaphnia dubia CdNaCICR #:

I b t" Day (Analyst identified for each day, performed PK-D.O. and conductivity measurements on l,.

3 4

5 6

4Analyst zPl orBMjoa Concentration Parameter pH (S.u.)

"-CS 7.S 1.S

. S

.5 7.i

't.

DO (mg*L) 77 0

7..

Conductivity (tunhos/cm)3o31

Alkalinity CONTROL

Aklinity (Ing CaCO3/L) t 2

Hardness (Ing CaC03/L)

Temperature 0.0

-o A 'L L Vi.A.0.*\\

.Lblu aAA DO (mg/L) 7.q S600 mg NaCIIL Conductivity I~)___

Temperature t j

'*z..

8, pH (S.U.)

.75

-7.'06 196

.2.1. 2.+/-*I

-z.

DO(mg/L)

.o

-.q 2

800mg NaCI/L Conductivityo

,o m

gmhos/cm

Temperature "q*

S LO "t,*

7:t L,

I) MY

_T_

pH (S.U.)

/1 -1P I.7.-77 I 7.93

'7.2 1 -7 It 1~ 1. -1. 1 A.n 7.7"L U *-'E~-4-'---- II 4 __________

4 4I-.L~~'---4 M.Z

.11 DO (ma/L)

W-4 11 1000 mg NaCI/L Conductivity r

(jImhostcm) 1

__6

_0 i

03O "U.0 u1.5

Temperature (00)

-0.1 11 -LS. 1) 11%S.% I pH (S.U.)

"7.2I

".Lq 7.7' 811."

".Z 7.7 DO (rag/L) 7=

)-*

7.

1200 mng NaCL/L Conductivity (ginhos/cmn) 9141

Temperature

.. PH (S.U.)

13--

-7*

/q 7.q1"j

{

DO (mg/L).

7.q 8.o 1400 mg NaCIIL Conductivity

Temperature

.,L.

Lq.*

-Let

-Lq.

"T.. 1 1

¢CIL Initial Final I-Initial 11 Final I Initial 11 Final I Initial 11 Final I

U a

r U

m

.0

Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1. Alkalinity and hardness performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by:

Z.

ge 99 of 99 SOP ATI4 - Exhibit ATI4.1, revision 06-01-11

ML12286A328

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ML12286A328

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Dear Ms. Morgan:

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