Text

Discharge Monitoring Report for August 2015
	ML15258A023
Person / Time
Site:	Sequoyah
Issue date:	09/10/2015
From:	John Carlin; Tennessee Valley Authority
To:	Morgan C; Office of Nuclear Reactor Regulation, State of TN, Dept of Environment & Conservation, Div of Water Resources
References
	TN0026450
	Download: ML15258A023 (102)
	v • d • e

Tennessee Valley Authority, Post Office Biox 2000, Soddy Daisy, Tennessee 37384.2000 September 10, 2015 Ms. Christina Morgan Tennessee Department of Environment and Conservation Division of Water Resources William R. Snodgrass Tennessee Tower 312 Rosa L. Parks Avenue, 11th Floor Nashville, Tennessee 37243

Dear Ms. Morgan:

TENNESSEE VALLEY AUTHORITY (TVA) - SEQUOYAH NUCLEAR PLANT (SQN) - NPDES PERMIT NO. TN0026450 - DISCHARGE MONITORING REPORT (DMR) FOR August 2015 Enclosed is the June 2015 Discharge Monitoring Report for Sequoyah Nuclear Plant. Toxicity at outfall 101 was sampled August 9 - 14, 2015. The toxicity report is enclosed. There were no exceedances during the monitoring period. If you have any questions or need additional information, please contact Millicent Garland by email at mrmoore@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 in formation, including the possibility of fine and imprisonment for knowing violations.

Enclosures cc (Enclosures):

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

PERMrITTEE NAMEIADDRESS (Include Facility Name/Location if Different)

Name TVA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 iNT~EROFCE OPS-5.N-SQ.N}_

)

SOD DY__-

DAISY. TN N.37384_

Fac~ili_...

A -S..EQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY ATTN:Milllcent Garland NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

DISCHARGE MONITORING REPORT (DMR)

It~65 MAJOR (SUBR 01)

Form Approved.

OMB No. 2040-0004 I

~MONITORING PERIOD F - FINAL DIFFUSER DISCHARGE EFFLUENT

- NO DISCHARGE

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No closed mode operation. The following injections occurred: Flogard MS6236 (max caIc. conc. 0.03 mg/L - limit 0.2 mg/L), Spectrus CT 1300 (max calc. conc. 0.03 mg/L - limit 0.05 mg/L)

EPA Form 3320-1 (REV 3199)

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

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

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SO DDY - DAISY, TN.37384 Faciity TVA - SQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY ATTN:Millicent Garland NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

MAJOR Form Approved.

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EPA Form 3320-t (REV 3i99)

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PERMIT:TEE NAME/ADDRESS (Include Facility Name/Location if Different)

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- - - - 2 DEG C CON'rIN CALCTD EFFLUENT GROSS VALUE REQUIREMENT DAILY MX uous SAMPLE MEASUREMENT PERMIT REQUIREMENT NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that thia document and all attachmenta ware prepared under my TELEPHONE DATE John T. Carlin peraonnel properly gather and evaluate the information aubmitted. Baaed on my inquiry of the peraon or peraons who manage the system, or those persona directly responsible tor gathering StViePeietthe information, the information aubmitted is. to the beat of my knowledge and belief, true, 423 843-7001 15 09 09 Sit VcePreidntaccurate, and complete, I am aware that there are significant penaltiea for aubmittirng false SIGNA'T R"E F PRINCIPAL EXECUTIVE information, including the possibility of fine and imprisonment for knowing violations.

OFFICER OR AUTHORIZED AGENT AREAI NUMBER YEAR MO DAY TYPED OR PRINTED

_____________________________CODE COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference al/ attachments here)

No Discharge this Period EPA Form 3320-I (REV 3/99)

Previous editions mat be used PaaP. 1 nf 1

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

Name TMA - SEQUOYAH NUCLEAR PLANT Address P.O. BOX 2000 IJNTEROFFICE OPS-hN-SCjN)

SODDY - DAISY, TN. 37384_

Facility TVA - SEQUOYAH NUCLEAR PLANT Location HAMILTON COUNTY A'l-N:Millicent Garland NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR DISCHARGE MONITORING REPORT (DMR)

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OFFI(*R-AUTHORIZED AGENT AREAI NUMBER YEAR MO DAY TYPED OR PRINTED

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

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

Previous editions maybe used Paoe 1 of 1

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the information, the information aubmitted is, to the best of my knowledge anid belief, true, eI~W 423 843-7001 15 09 09 Site Vice President accurate, and complete. l am aware that there are significant penalties for submitting false SIGN TU*F,*

PRINCIPAL EXEC UTIVE information, including the possibility of fine and imprisonment for knowing violations.

OFEFICE,0* AUTHORIZED AGENT AREAI NUMBER YEAR MO DAY TYPED OR PRINTED CD COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

During this reporting period, there has been no flow from the Dredge Pond other than-that resulting from rainfall. No Discharge this Period EPA Form 3320-1 (REV 3/99)

Previous editions may be used P*n* 1 nf t

TENNESSEE VALLEY AUTHORITY TOXICITY TEST REPORT INTRODUCTION / EXECUTIVE

SUMMARY

Report Date: September 03. 2015

1. Facility / Discharger: Sequovah 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): 1,5792

6. Receiving Stream: Tennessee River (TRM 483.6)

7.

1Q10: 3491

8. Outfall Tested: 101

9. Dates Sampled: August 09-14. 2015

10. Average Flow on Days Sampled (MGD): 1793. 1776. 1780

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

12. Test Dates: August 11 - 18,.2015

13. Test Type: Short-term Chronic Definitive

14. Test Species: Fathead Minnows (Pimeyhales promelas)

Daphnids ( Ceriodap~hnia dubia)

15. Concentrations Tested (%): Outfall 101: 10.7, 21.4,.42.8. 85.6. 100 Intake: 10__0 Pimephalespromelas: UV treated Outfall 101: 10.7, 21.4,.42.8. 85.6. 100 UV treated Intake: 10__0

16. Permit Limit Endpoint (%): Outfall 101:I1C25 = 42.8%

17. Test Results: Outfall 101: Pimephales promelas: IC_2

_ 5> 100%

Ceriodaphnia dub ia: IC25 > 100%

UV treated Outfall 101: Pimephales promelas: IC25 > 100%

Page 1 of 95

18. Facility

Contact:

Millicent Garland Phn#:(2)8361 Phone #: (423) 843-6714

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

20. Lab

Contact:

Jim Sumner Phone #I: (828) 350-9364

21. TVA

Contact:

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

22. Notes: Exposures to samples collected August 09 - 14, 2015 from Outfall 101 resulted in no toxic effects to fathead minnows or daphnids. The resulting IC25 values, for both spec~ies, were > 100 percent. Exposure daphnids to intake samples resulted in no significant difference from the control during this study period.

Growth was significantly reduced in minnows exposed to intake samples.

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 95

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

°)

(mg/L)

Last Used By 08-109-15 0600t 08-10-15 1205 1.8,1l.3t

<0.10 08-11-15 1121 101 08-109-15 0700 to08-12-15 1022 08-09-15 0720 to 08-11-15 1121 Intake 0-01060 08-10-15 1205 1.7

<0.10 081-5 02 08-12-15 0600 08-14-15 1022 08-11-15 0722 to 08-13-15 1021 Intak 08-12-15 0622 08-12-15 1300 1.2,13

<0.10 08-14-15 1022 08-1115 072 to08-15-15 1028 10 08-3-1ake0t 08-14-15 1306 1.27O.7

<0.10 081-5 02 08-14-15 0600 08-17-15 1021 08-15-15 1028 Intak 08-13-15 0718 to 08-14-15 1306 1.9,07

<0.10 08-16-15 1025 08-14-15 0618 08-17-15 1021

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 101 and intake were warmed to test temperature (25.0 +/- I1.0°C) in a warm water bath.

Aliqiuots 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 95

Pimephales prom elas Test Organisms:

Ceriodaphnia dub ia In-house Cultures

1. Source:

Aquatox, Inc.

2. Age:

18.17 - 19.35 hours4.050926e-4 days 0.00972 hours 5.787037e-5 weeks 1.33175e-5 months old

< 24-hours old Test Method Summary_:

1. Test Conditions:

Static, Renewal Static, Renewal

2. Test Duration:

7 d ays_

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: 1_f0

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:

1._00 1

08-11-15 1121 ET 08-11-15 1110OET 08-18-15 1035 ET 08-18-15 1017ET Mean = 24.7°C (24.2 - 25.2°C) 08-11-15 0900 ET 08-18-15 0803 ET Mean = 24.9°C (24.7 - 25.3°C)

Test Temperature: UV-Treated Outfall 101:

Mean =24.8uC (24.3 - 25.3°C)

9. Physical / Chemical Measurements:

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

10. Statistics:

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

Page 4 of 95

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 11 - 18, 2015 using effluent from Outfall 101.

Test Percent Surviving Solutions

(___

time interval used - days)

(% Effluent) 1 2

_J 3 4 [ 5 6

7 Control 100 100 100 100 100 100 100 10.7%

100 100 100 100 100 100 100 21.4%

100 100 98 98 98 98

.98 42.8%

100 100 100 100 100 100 100 85.6%

100 100 100 100 98 98 98 100.0%

100 100 98 98 95 95 95 Intake 100 100 100 100 100 88 88 TestSoluionsMean Dry Weight (mag)

TestSoluions(replicate number)

(% Effluent) 1 I

2 I

3

] 4 I¶ Mean Control 0.714 0.568 0.752 0.738 0.693 10.7%

0.655 0.742 0.691 0.707 0.699 21.4%

0.671 0.708 0.596 0.678 0.663 42.8%

0.607 0.659 0.660 0.790 0.679 85.6%

0.614 0.716 0.728 0.662 0.680 100.0%

0.762 0.644 0.707 0.662 0.694 Intake 0.527 0.576 0.545 0.635 0.571 IC25 Value: > 100%

Calculated TU Estimates: < 1.0 TUc*

Permit Limit: 42.8%

Permit Limit: 2.3 TUc 95% Confidence Limits:

Upper Limit: NA Lower Limit: NA

TUa = 100/LC5 0: TUc =100/IC25 Page 5 of 95

TOXICITY TEST RESULTS (see Appendix C for Bench Sheets)

(Genus species)

(Type / Duration)

Conducted August 11 - 18. 2015 using effluent from Outfall 101.

Percent Surviving Test

________(time interval used - days)_____

Solutions

(% Effluent) 1 2

3 4

567 Control 100 100 100 100 100 100 100 10.7%

100 100 100 100 100 100 100 21.4%

100 100 100 100 100 100 100 42.8%

100 100 100 100 100 100 100 85.6%

100 100 100 100 100 100 100 100.0%

100 100 100 100 100 100 100 TestSoluionsReproduction (#young/female/7 days)

Test Solutions

~Data_(replicate number)

(%Effluent) 11 2 3[I4 5 1_6 7

8 9]I10 IMean Control 30 30 29 28 29 30 33 31 30 31 30.1 10.7%

33 30 32 33 30 31' 32 30 31 30 31.2 21.4%

34 33 32 31 32 31 30 32 31 33 31.9 42.8%

34 33 35 30 31 32 31

33.

31 31 32.1 85.6%

35 34 35 34 35 33 37 34 34 35 34.6 100.0%

35 36 35 35 35 36 35 37 36 35 35.5 IC25 Value: > 100%

Calculated TU Estimates: < 1.0 TUc*

Permit Limit: 42.8%

Permit Limit: 2.3 TUc 95% Confidence Limits:

Upper Limit: NA Lower Limit: NA__________________

TUa = 100/LC5 0: TUc = 100/IC25

.Page 6of 95

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 11 - 18. 2015 using water from Intake Percent Surviving Test

________(time interval' used - dys)

Solutions 1

1 4

I 6

Control 100 100 100 100 100 100 100 Intake 100 100 100 100 1100 100 100 TestSoluionsReproduction (#young/female/7 days)

TestSoluionsData (replicate number)

(%Effiuent) 1 2

3 I4 j5 6 I7 I8 9

101 ~Mean Control 30 28 29 28 30 29 30 30 30 27 29.1 Intake 34 37 38 36 36 35 38 33 32 34 35.3 IC25 Value: > 100%

Calculated TU Estimates: < 1.0 TUc*

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

Upper Limit: N._A_

Lower Limit: NA

TUa =100/LCs0: TUc =100/IC 25 Page 7 of 95

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 11 - 18. 2015 using effluent from UV Treated Outfall 101.

Test Percent Surviving Solutions

________(time interval used -_days)___

(% Effluent) 11 2

[3 4

5 161 7

Control 100 100 100 100 100 100 100 10.7%

100 100 100 100 100 100 100 21.4%

100 100 100 100 100 100 100 42.8%

100 100 100 100 100 100 100 85.6%

100 100 100 100

.100 100 100 100.0%

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

Test Solutions (replicate number)

(% Effluent) 1

[

2 3

[

4 j¶ Mean Control 0.785 0.659 0.655 0.747 0.712 10.7%

0.648 0.588 0.726 0.724 0.672 21.4%

0.712 0.767 0.619 0.824 0.731 42.8%

0.655 0.738 0.747 0.753 0.723 85.6%

0.811 0.803 0.815 0.715 0.786 100.0%

0.737 0.713 0.869 0.809 0.782 Intake 0.694 0.743 0.689 0.670 0.699 IC25 Value: > 100%

Calculated TU Estimates: < 1.0 TUc*

95% Confidence Limits:

Upper Limit: N__AA Lower Limit: NA

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

Species Date Time I Duration I Toxicant I Results (IC25)

Pimephalespromelas August 11 - 18, 2015 1100 7-days KC1 0.81 g/L Ceriodaphnia dubia August 04 - 11 2015 0935 7-days NaC1 1.06 g/L*

Page 8 of 95

0 01 PHYSICAIJCIIEMICAL

SUMMARY

Water Chemisty Mean Values and Ranges fir Pimephalespromelas and Ceriodaphnia dubia Tests, Non-hreated SequoyahNuclear Plant (SQN) OutJ~l1 101 performed August 11-18, 2015.

Test Sample ID Temperature (°C)

Dissolved Oxygen (rag/L) pH (S.U.)

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

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

Chlorine (rag/L)

Cnrl 24.8 24.5 7.8 7.7 7.75 7.52 318 60 86 24.7-24.9 24.3 - 24.7 7.6 7.9 7.0 8.0 7.63 -

7.98 7.39-7.68 305 -

327 59 61 84

-88 0.%

24.8 24.4 7.8 7.7 7.74 7.48 301 24.8 - 25.0 24.2 -

24.6 7.7 7.9 6.9 8.0 7.67 -

7.79 7.35 -

7.67 291 315 2.%

24.9 24.5 7.9 7.7 7.74 7.46 287

_____24.8

- 25.0 24.3 - 24.7 7.8 8.0 6.9 8.0 7.68 -

7.78 7.32 - 7.65 280 -

301 43.2%

24.9 24.5 7.9 7.8 7.74 7.47 258 24.8 - 25.0 24.3

- 24.6 7.8 8.0 7.0 8.1 7.68 -

7.80 7.29 - 7.65 253 263 8.%

24.9 24.5 7.9 7.8 7.73 7.45 206

______24.8 25.0 24.3

- 24.7 7.8 8.1 6.9 8.1 7.67 - 7.79 7.23

- 7.68 200 -

209 10.%

24.9 24.6 7.9 7.7 7.73 7.46 187 70 73

< 0.10

_____24.8

- 25.1 24.3 - 24.7 7.8 8.1 6.5 8.1 7.67 - 7.80 7.20

- 7.69 181 192 67

-73 72

-76

<0.10 -

<0.10 Inae 24.9 24.5 8.0 7.8 o 7.73 7.55 189 69 72

< 0.10 24.7 -

25.2 24.3

- 24.6 7.8 8.1 6.9 8.1 7.67 - 7.79 7.41

- 7.70 184 -

191 65

-73 72 72

<0.10 -

<0.10 Cnrl 24.8 25.0 7.8 7.9 7.75 7.67 318 60 86

_____24.7 24.9 24.8 - 25.2 7.6 7.9 7.6 8.1 7.63 - 7.98 7.54 - 7.77 305 -

327 59

-61 84 88 108.

24.8 25.0 7.8 7.9 7.74 7.69 301 24.7 -

24.9 24.8 -

25.1 7.7 7.9 7.6 8.1 7.67 - 7.79 7.62 - 7.76 291 315 21.6%

24.9 25.0 7.9 7.9 7.74 7.69 287 216%

24.7 -

25.0 24.8 - 25.2 7.8 8.0 7.6 8.1 7.68

- 7.78 7.63 - 7.76 280 -

301 42%

24.9 25.0 7.9 7.9 7.74 7.69 258 4"°0 24.8 -25.0 24.8 -25.3 7.8

-8.0 7.7

-8.1 7.68

-7.80 7.63 -7.80 253

-263 84%

24.9 24.9 7.9 7.9 7.73 7.70 206

_____24.8

- 25.0 24.8 - 25.0 7.8 8.1 7.8 8.1 7.67 - 7.79 7.63 -

7.81 200 -

209 10.%

24.9 25.0 7.9 7.9 7.73 7.70 187 70 73

< 0.10 10.°0 24.8 - 25.0 24.8 -

25.1 7.8 8.1 7.8 8.1 7.67 - 7.80 7.63 - 7.83 181 192 67

-73 72 76

<0.10 -

<0.10 Inae 24.9 25.0 8.0 8.0 7.73 7.72 189 69 72

< 0.10

____24.8 25.0 24.9 -

25.1 7.8 8.1 7.8 8.1 7.67 - 7.79 7.44 - 7.83 184 -

191 65

-73 72 72

<0.10 -

<0.10 Overall temperature (eC Pimephales promelas Ceriodaphnia dubia Average 24.7 24.9 Minimum 24.2 24.7 Maximum 25.2 25.3

tol U'

PHVSICAIJCttEMICAL

SUMMARY

Water Chemistry Mean Values and Ranges forPimephalespromelas Test, UV'-treated SequoyahNuelear Plant (SQN) Ouffal1 101 peribrrr~d August 11-18, 2015.

Test Sample ID Temperature ("C)

Dissolved Oxygen (rag/L) pH (S.U.)

Conductance Alkalinity Hardness

Total Residual Initial Final Initial Final Initial Final

@tmhos/cm) (mg/'L CaCO 3) (mg/L CaCO 3) Chlorine (mg/L)

Cnrl 24.9 24.5 7.8 7.7 7.74 "

7.57 316 60 84 24.8 - 24.9 24.3 -

24.7 7.7 8.0 6.9 8.0 7.68

- 7.80 7.33 7.71 307 -

327 59

-60 82 86 0.%

24.9 24.5 7.9 7.7 7.74 7.53 304 24.8 - 25.0 24.4 -

24.7 7.8 8.0 6.9 8.1 7.59 - 7.84 7.32 -

7.71 294 -

316

-25.0 24.6 7.9 7.7 7.76 7.53 292 21.6%

43.2%

25.0 24.5 7.9 7.7 7.77 7.51 262

_____24.8

- 25.1 24.3 -

24.7 7.8 8.0 7.0 8.1 7.68 - 7.86 7.27 -

7.71 255 -

269 86.4%

25.0 24.6 7.9 7.7 7.77 7.51 209 24.8 - 25.1 24.3 -

24.7 7.8

-8.1 7.0 8.1 7.69 - 7.87 7.28 -

7.71 202 217 10.%

25.1 24.4 7.9 7.7 7.78 7.50 191 69 73

< 0.10

_____24.9

- 25.3 24.3 -

24.6 7.8

-8.1 6.9 8.1 7.70 -

7.88 7.28 7.71 183 198 67 71 72

-76

<0.10 -

<0.10 Inae 25.1 24.5 8.0 7.7 7.79 7.56 191 69 72

< 0.10

____25.0

- 25.2 24.4 - 24.6 7.8

-8.1 6.6 8.1 7.70 -

7.88 7.39 - 7.71 183 203 67 71 72

-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 Pimephalespromelas 24.8 24.3 25.3

SUMMARY

/ CONCLUSIONS Exposures to samples collected August 09 - 14, 2015 from Outfall 101 resulted in no toxic effects to fathead minnows or daphnids. The resulting IC2s values, for both species, were > 100 percent. Exposure daphnids to intake samples resulted in no significant difference from the control during this study period. Growth was significantly reduced in minnows exposed to intake samples.

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 95

Appendix A ADDITIONAL TOXICITY TEST INFORMATION

SUMMARY

OF METHODS

1. Pimephales promelas Tests were conducted according to EPA-82 1l-R-02-0 13 (October 2002) using' four replicates, each containing ten test organisms, per treatment. Test vessels consisted of 500-mL plastic disposable cups, each containing 250-mL of test solution.

2. Ceriodaphnia dubia Tests were conducted according to EPA-82 1-R-02-0 13 (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 I MODIFICATIONS TO TEST PROTOCOL

1. Pimephales promnelas None

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

1. Pimephales promelas None

2. Ceriodaphnia dubia

.None Page 12 of 95

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-2000.

4. Dissolved oxygen was measured by SM 4500-0 G-2001.

5. The pH was measured by SM 4500-H+ B-2000.

6, Conductance was measured by SM 2510 B-1997.

7, Alkalinity was measured by SM 2320 B-1997.

8, Total hardness was measured by SM 2340 C-1997.

9, Total residual chlorine was measured by ORION 97-70-1977.

.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 IC25 values in g/L KCI or NaC1.

2.

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

Sodium Chloride (NaC1 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 95

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-821l-R-02-013 (October 2002).

3. Standard Methods for the Examination of Water and Wastewater. 2 2nd Edition, 2012.

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

Page 14 of 95

Sequoyah Nuclear Plant Biomonitoring August 11 - 18, 2015 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 95

Table B-i1. Sequoyah Nuclear Plant Outfall 101 Diffuser Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion and Mollusks, During Toxicity Test Sampling, February 6, 2005 -August 14, 2015

Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-PF H-130M Nalco 1H-150M Hypochlorite mgiL mg/L mg/L mg/L mg/b mg/b 73551 mg/b mg/b TRC Phosphate Copolymer DMAD)

Azole Quat mg/L Quat TRC

____EO/PO J

02/06/2005

<0.0042 0.028 0.010-----

02/07/2005

<0.0116 0.028 0.010 0.007-02/08/2005

<0.0080 0.028 0.010-----

02/09/2005

-0.0199 0.028 0.010-----

02/10/2005

<0.0042 0.028 0.0 10-----

02/11/2005

-0.0155 0.028 0.010 0.007-06/05/2005

-0.0063-------

06/06/2005' 0.0043 0.037 06/07/2005

-0.0103 0.037 06/08/2005

-0.0295 0.037 06/09/2005

-0.0129 06/10/2005

-0.0184-------

07/17/2005

-0.0109 0.026 0.009-07/18/2005

-0.0150 0.026.

0.009

0.036 07/19/2005

-0.0163 0.026 0.009 0.036 07/20/2005

-0.0209 0.026 0.009 0.014 0.036 07/21/2005

-0.0242 0.026.

0.009 07/22/2005

-0.0238 0.054 0.018 0.014-10)/30/2005

-0.0068 10/31/2005

-0.0112------

11/01/2005

-0.0104 0.035 11/02/2005

-0.0104 0.036 11/03/2005

-0.0117 0.036 11/04/2005 0.0165 0.035 11/14/2005

-0.0274-------

11/15/2005

-0.0256-------

11/16/2005 0.0234 11/17/2005

-0.0231 11/18/2005

-0.0200-------

11/19/2005

-0.0116-------

Page 16 of 95

Table B-i1. Sequoyah Nuclear Plant Outfall 101 Diffuser Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion and Mollusks, During Toxicity Test Sampling, February 6, 2005 -August 14, 2015 f Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-PF H-130M Nalco H-150M TMSW mg/b TRC Phosphate Copolymer DMAD Azole Quat mg/b Quat mg/L

_ J TIC

_____EO/PO IPhosphate 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

-0.023 8

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 "1

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 17 of 95

Table B-1. Sequoyah Nuclear Plant Outfall 101 Diffuser Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion and Mollusks, During Toxicity Test Sampling, February 6, 2005 -August 14, 2015 Date Sodium Towerbrorm PCL-PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSW Hypochlorite mg/b 222 mg/b mg/b

-PF mg/L mg/b 73551 CT*1300 mg/L 101 mg/C Phosmgh Cooye MD Aoe Qa Eg/LO Quat Phosphate mgbRRCm/

Coopoh me DAEAoiPuaOm/

mg/b Quat pmg/b

_____________ate

_____j_____

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 18 of 95

Table B-i. Sequoyah Nuclear Plant Outfall 101 Diffuser Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion and Mollusks, During Toxicity Test Sampling, February 6, 2005 - August 14, 2015 Date Sodium Towerbrom PCL-PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSW Floguard Hypo-mg/L 222 mg/b mgiL

-PF mg/b mg/b 73551 CT1300 mg/b 101 MS6236 chlorite TRC mg/b Copoly-DMAD Azole Quat mg/b mg/b Quat mg/b mg/b mg/b Phos-mer 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 0.04 05/05/2011 0.0179 0.04' 05/06/2011 0.0089 11/06/2011 0.0168 11/07/2011 0.0225 11/08/2011 0.0141 11/09/2011 0.0239 11I/10/2011 0.0242 11I/11/2011 0.0231 05/06/2012 05/07/20 12 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 08/15/20 12 0.0279 0.028 0.029 08/16/2012 0.0076 0.029 08/17/2012 0.0446 0.032 05/12/20 13 0.0099 05/13/2013 0.064 05/14/2013 0.0091 0.039 0.064 05/15/2013 0.0096 0.039 0.064 05/16/2013 0.0229 0.032 05/17/2013 0.0063 0.032 09/15/2013 0.03 09/16/2013 0.0072 0.0379 0.03 09/17/2013 0.0107 0.036 0.0379 0.03 09/18/2013 0.0217 0.036 0.0379 0.03 09/19/2013 0.0172 0.03 09/20/2013 0.0173 0.03 Page 19 of 95

Table B-1. Sequoyah Nuclear Plant Outfall 101 Diffuser Discharge Concentrations of Chemicals Used to Control Microbiologically Induced Corrosion and Mollusks, During Toxicity Test Sampling, February 6, 2005 -August 14, 2015 Date Sodiumr 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/b mg/b 73551 CT1300 mg/b 10,1 MS6236 chlorite TRC mg/b Copoly-DMAD Azole Quat mg/b mg/b Quat mg/b mg/b mg/b Phos-mer EO/PO Quat Phos-Phosphate TRC phate

____phate 05/04/2014 0.0118 05/05/2014 0.0112 05/06/20 14 0.0096 05/07/2014 0.0164 05/08/2014 0.0235 05/09/2014 0.0110 09/07/2014 09/08/2014 0.04---

09/09/2014.

0.04 09/10/2014 0.04 09/11/2014 0.0070 09/12/2014 0.0074 08/09/2015 08/10/2015 0.0195 0.03 08/11/2015 0.0275 0.03 08/12/2015 0.0213 0.03 08/13/2015 0.0192 0.03 08/14/2015 0.0182 0.03 Page 20 of 95

Sequoyah Nuclear Plant Biomonitoring

Augustl11-18, 2015 Appendix C Chain of Custody Records and Toxicity Test Bench Sheets Page 21 of 95

lient: TVA Environmental Testing Solution, Inc.

351 Depot Street.

Name: Sequoyah MP Toxicity

.Number: Nq/A Asheville, NC 28801 Sampled: Sequoyah NP Phone:

828-350-9364 NPDES Number: TN0026450 EcA i5ac~4

~

LA~O~S ~

to

~vd&~

~o I uS' ov~

Fax:

828-350-9368 Collected Sample Description

.*-1IJ l-I Instructions: Clients should fill in all areas except those in the "Laboratory Use" block. Bionionitoring samples are preserved by storing them at 60C and shipping them in ice. The hold time for each sample is 3 6 hours6.944444e-5 days 0.00167 hours 9.920635e-6 weeks 2.283e-6 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 ovteright on Friday vi'a FedEx or UiPS must be marked for Saturday delivery or they will not arrive until the following Monday.

Whole Effluent Sample Receipt Log Page__*

Sample temperatuze performed using Sample Rereiung Thermoameten~ SN 130580151 O

-Ii

'I' SOP 64 - Exhibit 64.2, revision 0 1-03-12

BIOMONITORING CHAIN OF CUSTODY RECORDPae1o1 Page 1

of 1

U U

~ient: TVA...

roject Name: Sequoyah NP Toxicity

.O.Number: N/A

~iiiySampled: Sequoyal,,NP NPDES Number: TN0026450 Co-let.

Environmental Testing Solution, Inc.

351 Depot Street.

Asheville, NC 28801 Phone:

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

FedEx UPS Bus Client Other(specify):

Sc:r'xi. ?

_[/¢ General Comments:

i Fax:

828-350-9368 Field Ietfcto Sample Description ColcinDt/ie CNtainer Fo SQN-101-TOX Comp X/tt gig/

2 (2.5Sgal)

Sample Custody - Fill In From Top Down

-it. L¢*mi-1 sa*.

1t*,N 6A*&0*S

tI*_A.'t*.1 RlnuseBy(Signature):

Date/Time Received By (Signature):

¢sbCt*a Date/Tunet

____~

S

~

f3 i,

iS 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 bold time for each sample is 36 hours4.166667e-4 days 0.01 hours 5.952381e-5 weeks 1.3698e-5 months front 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.

m II

Page **

Whole Effluent Sample Receipt Log

Sampletemperstura performed using Sample ReceivngmTermometer: SN 1305801.51 U'

0

-6D Date lime

Received Received

Sample Project Sample Received Received by from Temp. (°e) number number Sample name and description

State, Comments 08-12-15 1002 K. Keenan Fed - Ex 0.5 10848 150812.01 ALCOA - 005 NC 08-12-15 1002 K(. Keenan Fed - Ex 0.6 10849 150812.02 Apex WRF.

NC

.08-12-15 1002 K. Keenan Fed - Ex 2.7 10850 150812..03

Daikin Ap~pl~ied Americas, Inc.

NC 08-12-15 1002 K. Keenan Fed - Ex 0.7 10851 150812.04 Dallas WWTP NC 08-12-15 1002 K. Keenan Fed - Ex 0.5 10852 150812,.05 McGuire NS -005 NC 08-12-15 1002 K. Keenan Fed - Ex 0.4 10853 150812.06 Craven County Wood Energy NC 08-12-15 1002 K.K Keenan Fed - Ex

0.7 10854 150812.07 Elementis NC 08-12-15 1002 K. Keenan Fed - Ex 1.1 10855 150812.08 Wood lake Yacht Club NC 08-12-15 1002_

K. Keenan Fed - Ex 0.7 10856 150812.09 MeGuire N*S

- 00:1 NC 08-12-15 1002 K. Keenan Fed - Ex 1.7 10857 150822.10 McGuire NS -002 NC 08-1!2-15 1300 3J.

Sumner TVA Courier 1.5./.1.3 10842 "150812

.11 TVA - Sequoyah.Nuclear Plant -,101 TN' 08-12-15

.1300

1. Sumner TVA Courier 1.2.

10842 150812.12 TVA - Seguoyah Nuclear Plant - Intake

TN SOP G4 - Exhibit G4.2, revision 01-03-1:2

ffIOMONflTORING CHAIN OF CUSTODY RECOR.LD Page_ 1_of 1

,tlent: TVA Environmental Testing Solution, Inc Delivered By (Circle One):

¢Dlloject Name: Sequoyah NqP Toxicity 351 Depot Street.

FedEx UPS Bus Client

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

L),(o *l'Zt*

mclt Sape:Sqoa P2880 1

General Comments:

NPDES Number: TN002645O Phone:

828-350-9364

.TjIe%

bi. -

a@o J

.,+d j*r'OI 77 Coil~

~'

Fax:

828-350-9368

- 5a*.*,

Field Identification /

t/Comp

]lolctn ae/meC tier Flow A*<.*:e+.,-l1Ea-+ie-.o 1

++I:

Sanmple Description Number &

(M GD)

Rain Event?.

,@.*+

+:*..

%:+;' :;,*":

Volmne

~~(Mark as Appropriate)

t"'

ba,-: at.oi"y-':Us-e:+

3:

YesecedI f Yes, No Trace

-.A r n.V:: Tmp

-B""'

":"it:e Appear SQNAOI-TOX Co,,,pn'

,(.g2<+a)>

,4.[7<

i::+'

2-

...... i SQN-INT-TOX Comp

'-13-I5" W-I'-b-J 1 (2.5 gal)

'NA I

V

+.*::=,&

L !+;::

Sample Custody - Fill In From T*op Down

,mb t,.Jt'*

4 S

tJ-Pt

+*!,.

9~4A

.e t

a

cI Relinquished By (Signature):

Date/Time Received By (Signature):

.tatlim 7-*

.ol 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 k(e. 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.

@IT Page____.

Whole Effluent Sample Receipt Log

Sample temperature performed using Sample Receiving Thermometer: SN 130580151 0

0I CO "Date Time Received "Received

Sample Project Sample Sapl.nmean.dsc..io..a..C.mnt Received Received by from Temp. (0C) number number 0.8-1,4-15 0950 K. Keenan Fed - Ex 0.5 10848 150814.01 ALCOA - 005 NC 08-14-15 0950 K. Keenan Fed - Ex 1.6

10849 150814.02 Apex WRJF N.C 08-14-15 0950 K. Keenan Fed - Ex 1.1 10850 150814.03 Dalkini Applied Americas, Inc.

NC 08-14-15 0950 K. Keenan Fed - Ex 1.0 10851.

150814.04 Dallas WW,,P TiC 08-14-15 0950.

K. Keenan Fed - Ex 0.7 10852 150814.05 Mc:Guire NS -005

,NC, 08-14-15 0950 K. Keenan Fed - Ex 1.0 10853 150814.06 Craven County Wood Energy NC 08-14-15 0950 K. Keenan Fed - Ex 2.0 10854 150814.07 Elementis NC 08-1L4-15 1306 J1. Sumner TVA Courier 1.7/0.7 10842 150814.08 WTA - Seguoyah Nucl~ear Plant - 101 TN 0.8-14-15 1306 J. Sumner TWA Courier 1.9 10842 150814.09 TWA - Seguoyah Nudlear Plant - Intake TN SOP G4 - Exhibit 64.2, revision 01-03-12

Page 1 of 6 Chronic Whole Effluent Toxicity Test. (EPA-82:I-R-02-013 Method 1000.0)

Species: Pimeo~hals oromelas Client: Tennsse!ieVaeyAuthority Facility: Sequovah Nuclear Plant NPDES #: TN0026450 Project #:

1t *;'&..

County: Hamilton Outfiulh 101.

Dilution preparation information:

Contments:

Dilution prep (%)

10.7 21.4 [42.8 85.6 100' E. ffluent volume (mL),.

267.5 535 1070 2140 2500 Diluent volume (mL) 2232.5 1965 I1430 I360 0

Total volujne (niL) 2500 2500 2500 2500 2500 Test organism information:

j..

Tetinformation:J Orgismage t*] %x*

'AK,*5

wt s Randmzgtepte n..~ n-nfl nYnPcnrnon1~rn~

I

.nn

.tar~

n-...w K,nnhlflatflr numnpr unir 1

-I an~,nzao t~=bL*n~wnAa were born between:

shelf location:

Organism source:

"ATOX Batch Pp: Ot.4*-6 Antemia CHM number:.j CHIM836 Drying inform ation for weight

_______________________determination:

Transfer bowl information:

pH

-=

S.U.

Date / Time in oven:

G&. tltI, Temperatre_=_______°C Jnitial oven temperature:

t**,.

Average transfer volume:

0.1749 mL Date I Time out of oven:

.t]

Final oven temperature:

t

___Total drying time:

Daily feeding and renewal information:

Day Date Morning feeding Afternoon feeding Test initiation, Sample numbers used MHtsW renewal, or batch termination used Time Analy*st Time Analyst

.Time

.Analyst

,Outfall 101 Intake S

6 08-17-15 08-18-15 a *\\I AL a

Control information:

Acceptanc orter a Summary of test endpOints:

% Mortality.

Ot. -

o2%

7-day" LCSB Average weih per initial larvae:

0 NOEC TU'.

Average weight per surviving larva~e:.0 AA°t

> 0.25mg/larvae LOEC

) I D7.

ChV

~' Ic~7.

IC25

[

>roO1.~

SOP AT20 - Exhibit AT20.3, revision 11-01-14

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Page 3 ofr6 Species: Pimevheles vromelas Client: TVA / Sequoyah Nuclear Plant. Outfall 101. NON-treated Date: 08-11-15 Survival wad Growth Data Day 42.8%

85.6%

-100%

M N

O 0 P

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S T

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5diI91 IS-&I I

5.t B *Pan + Larvae weight (rag) r~

Date:

e* **t-C= Larvae weight (rag) =B-A Analyst:

Weight per initial number of larvae (nmg)

Analyst:. _

9 D"

0"

(*° th" 1° Average weight per Percent reduction larvae (nag) ig unusually large, d&r = decanted and returned, w = woun~ded.

C mme......s:.

SOP AT20 -Exhibit AT20.3, revision 11-01-14

S6 ;o !,£ efed 417-1O-1I UO~s!^OJ 'v'OzJiv !q~'px

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d l

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'p~op = p.'.xuIp = o :sopoo vxuuommo 1.L l

1$' 0

(%)

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'I' TVA / Sequoyah Nuclear Plant, Outfail 101 Non-treated August 11-18, 2015 Pimephaleapromelas Chronic Wbole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Project number.

10842 C'otarwtesllae%)

Itllsj htalunlaluberel )IncabJez Ieolse A=Fae..elgbt (erg)

=P~an+L~rr~e 1~cawedgbt(ess Welsht/Saevlving MeawrelgitI Coedtekate~tetfvarI We~gh!tlettllianmber Memansarelost leMtedght/lalsth Ca!Ietcktef hecetlieau4fmer jear..

wetghtelsn

=A-B nnmferretn'g~)] Suan gimanulrerof er*~

of larne(erg

(%1 rnberoxflerva.,ee tlrhte Cseoendh ronfrol (%)

A 10 10

'16.10 "23.24

'7.'14 0.714

-0.714

t otrl B

10 10 16.08 21.76 5.68 0.568

0.693 12.2 0.7528-100 06312Ntapial Ctrl C

10 I 0_____

15.75 23.27 7*52 0.752 10.0.735222e plial D

10 10 16.19 23.57 7.38 0.738 0.738.

E" 10 10 12.92 19;47 6.55 0.655 0.655 0.%

F

,10 10 16.84 24.26 7.42 0.742 0.699 5.2 069210.

.9 62-.

107 010 16.11 23.02 6.91 0.6910.9 0.7078______

214 0

10 16.06 22.02 5.96 0.596

.630.70

7.

.637.

M10 0

16.45 22.52 6.07 0.607 0.607 2.% N 0'10 15.65 22.25

.6.50 0.660 0.679 11.5 0.659 100.0 0.679 11.5 2.0 P

1010 6.10 24.0 7.0 07900.790

______10 9

51 12

.4060.614

__,6__R 1010 15.58

..22.74 7.16 0.716 0.697.

4.4 0.716 97.5 0.680 7.7 1.9 S

10 10 15.17 22.45 7.28 0.728 0.728 T

10 10 15.15

.21.77 6.62 0.662 0.662...

U 10 10 14.95 22.57 7.62 0.762 0.762 100%

V 10 10 15.85 22.29 6.44 0.644 0.732 8.5 0.644 95.0 0.694 7.6

.0.1 W

10 9

14.94 22.01 7.07 0.786 0.707 X

10 9

15.81!

22.43 6.62 0.736 0.662 Y

10 7

15.80 21.07 5.27 0.753 0.527 100% Intakse Z

10 9

15.19

.20.95 5.76 0.640 0.658 9.057 87.5 0.571 8.3 17.6 AA 10

____9 14.32 19.77 5.45 0.606 0.54.5 EB 10 10 16.09 22.44 6.35 0.635 0.6.15 Ootfat1101:

D)unnett~n 8851 value:.

PMSD:

Daunnattra88SD valuee PMSD1:

.0.1041.

15.0 0.0944 13.6 0451 =

Mininrumn Significant Diffoerenc PMSD -"

Per'cent Minimmum Signif~cunt Difference PMSD is a teasaren ofltest precision. The FMSD is the minimum percent difference between the cont'ol and treatment that can be declared staitically sianillcant in a whole efflluent toxicity lest.

Lower PMSD boend determidned by USEPA (10th percentile) - 12%.

Upper 9045D bound determrined by USEPA (90th percentlile) = 30%.*

Lower and upper PMSD bounda were determined fines the 10th and 90th perenotile, respectively, of" PMSI3 data froas EPAas WET Interlabrar-ory Variability Study (USEPA, 2001 a USEPA, 2001 b).

UdSEFSA. 2001a, 2U00T. Final Report: Interlaboratory Variability Study of EPA Shor-~tcem Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes 1 and 2-Appendirc, EPA-82 1-B-01-004 end EPA-821-B-01-005. US Environmental Protection Agency. Cincinnati, OH.

Ftle: sqnl0O.08 t115d ata*xMsx' Entered by:. J. Spmnrm~

Reviewed by:..

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated August 11-18, 2015 Statistical Analyses Er C

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

ijrsi*.'J Test ID:

PpFRCR Sample ID:

"rVA/I SQN, Outfall1101 EndDat:

Lb I:

ES-rwi. Tst~g St.

Sample Tpe:

DMR-Disctnarge Monitorlng Report SSample Date: *s-~t-2d15 Protocol: FWCHR-EPA-821-R-02-013 Test Speciea:

PP-Pimephales promelaa Conmments:

Non-treated Conc-%

t 3

4 D-Control 0.7140 0.5680 0.7520 0.7380 10.7 0.5550, 0.7420 0.69*10 0.7070 21.4 0.6710 0.7080 0.5960 0.6780 42.8 0.6070 0.6590 0.6600 0.7900 86.6 0.6140 0.7160 0.7280 0.8620 100 0.7620 0.6440 0,7070 0.6620

Intake 0.5270 0.5760 0.5450 0.6350 Transform: Untransfornied 1-Tailed Isotonic Conc-%

Mean N-Mean Mean Mmn Max CV%

N t-Stat Critical MSD Mean N-Mean D-Control 0.8900 1.0000 0.6930 0.5500 V.7520 12.236 4

0.6959 1.0000 10.7 0.6988 1.00.83 0.8968 0.6550 0.7420 5.168 4

-0.133 2.410 0.1041 0.6959 1.0000 21.4 0.6633 0.9571 0.6633 0.5960 0.7080 7.180 4

0.669 2.410 0.1041 0.6790 0.9767 42.8 0.6790 0.9798 0.6790 0.6070 0.7900 11.492 4

0.324 2.410 0.1041 0.6790" 0.9757 85.6 0.6800 0.9812 0.6800 0.8140 0.7280 7.726 4

0.301 2.410 0.1041 0.8790 0.9757 100 0.6938 1.0011 0.6938 0.6440 0.7620 7.590 4

-0.017 2.410 0.1041 0.6790 0.9757 Intake 0.8708 0.8236 0.5705 0.5270 0.6360 8.300 4

Auxiliary Testa StatistIc Cditical Skew Kurt Shrapiro-Wiik's Test indicates normal distribution (p > 0.01) 0.98441 0.884

-0.2563 0.05031 Bartlett's Test indilcates equal variances (0 = 0.75) 2.65818 16.0863 Hypothesis Teat (1-tall. 0.05)

NOEC LOEC ClhV TU MSDu MSDp

MSS, MSE F-Prob df DunnettasTeat 100

>100 1

0.10409 0.1502 0.00069 0.00373 0.98457

5. 18 Treatments vs D-Controi Linear Interpolation (280 Resamples)

Point SD 95% CL(Exp)

Skew IC05

>100 ICIO

>100 1C15

>100 1.0 IC20

>100 u.9 IC25

>100 1C40

>100 0.8 1050

>100 0.7

== 0:.6 0.5.

0.4.

o.31 0.21 0.11 0.0

-°1*

-0,1 5O8 100 150 Dose %

Dose-Response Plot 1-tail, 0.05 level of significance Ind.r.,d.y re~v.wbv v.11ev r. b..,.n Page 33 of 95 File: eqnl01..08111I5deta~xlsx Ontered by: I. 5~nmrer Reviewed by.

TVA / Sequoyah Nuclear Plant, Outfall1101 - Intake Non-treated August 11-18, 2015 "ET I*

Envinronntalra! *tlng Solutiont, Inc, Statistical Analyses Larval Fish Growth and Survival Test-7 Day Growth Start Date:

aV*:..i:'*./

Test iD: PpFRCR Sample I D:

TVA /SQN, Outfall1101-Intake End Date:

Lab ID:

ETS-Envir. Testing Sal.

Sample Type:

DMR-Dlscharge Monitoring Report Sample Date: Auut 01 Proto*:9l: FVWCHR-EPA-821-R-02-013 Test Species:

PP-Pimepheles promelas Comments:

Non-treated Conc-%

1 2

3 4

D-Control*

0.7140 0.5680 0.7620 0.7380 10.7. 0.6550 0.7420 0.6910 0.7070 21.4 0.6710 0.7080 0.5960 0.6750 42.8 0.6070

'0,6590 0.6600 0.7900

85.6 0.6140 0.7160 0.7280 0.6620 100 0.7620 0.6440, 0.7070 0.6620 Intake 0.5270 0.5760 0.5450 0.6350 Transforrn: Untranstormed 1-Tailed Conc-%

Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD D=Control 0.6930 1.0000' 0.6930 0.5680 0.7520 12.236 4

10.7 0.6988 1.0083 0.6988 0.6550 0.7420 5.168 4

21.4 0.6633 0.9571 '0.6633 0.5960 0.7080 7.180 4

42.8 0.6790

0. 9798 0.6790

.0.6070 0.7900

11.492 4

85.6 0.6800 0.9812 0.6800-0.6140 0.7280 7.726 4

100 0.6938 1.0011 0.6938 0.6440 0.7620 7.590 4

Intake 0.5708 0.8236 0.5708 0.5270 0.6350 6.300 4

2.517 1.943 0.0944.

Auxiliary Tests Statistic Critical Skew Kurt Shaepiro-Wilk's Test indicates normal distribution (p > 0.01) 0.90738 0.749

-1.0985 1.0071 F-Test indicates equal variances (p = 0.36) 3.20404 47/.4683 Hypothesis Test (I-tail, 0.05)

-MSDu MSDp MSB MSE. Ff-Prob df Homoscedastic t Test indicates significant differences 0.09437 0.13618 0.02989 0.00472 0.04546 1., 6 Treatments vs D-Control Dose-Response Plot.

0.9 0.8 0.7

0.6 2 0.5 0.4

0.3 0.2 0.1 1-tail, 0.05 level of significance Pag 34 of 95 Pile: sqnlO1._Og I lSdata.xlsx Entered by: J. Sumner Reviewed by:

TVA / Sequoyah Nuclear Plant, Outfall 101 - Intake Non-treated August 11-18, 2015 SEnvlmnmnualTesting Solutions. In c.

Statistical Analyses

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

jEji:*

Test ID:

PpFRCR Sample ID:

TVA / SQN, Otll101 -Intake End Date:

Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date:

o*',4.

Protocol: FWCHR-EPA-821-R-02-013 Test Species:

PP-Pimephaies promelas Comments:

Non-treated.

Cone-%

1

2.

3 4

D-Control 1.0000 1.0000 1.0000 1.0000 10.7 1.0000 1.0000 1.0000 1.0000 21.4 1.0000 0.9000 1.0000 1.0000 42.8 1I.0000 1.0000 1.0000.

1.0000 85.6 0.9000 1.0000 1.0000 1.0000 lop 1.0000 1.0000 0.9000 0.9000 Intake 0.7000 0.9000 0.9000 1.0000 Transform: Arcsin Square Root 1-Tailed Conc-% '

Mean N-Mean Mean Min Max 011%

N t-Stat Critical MSD D-Control 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4

10.7 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4

21.4 0.9750 0.9750 1.3713 1.2490 1.4120 5.942 4

42.8 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4

85.6 0.9750 0.9750 1.3713 1.2490 1.4120 5.942 4

100 0.9500 0.9500 1.3305 1.2490 1.4120

.7.072 4

intake 0.8750 0.8750 1.2253 0.9912 1.4120 14.199 4

2.146 2.353 0.2047 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.78118 0.749

-0.8127 3.70059 Equality of variance cannot be confirmed Hypothesis Test (1-taiI, 0.05)

MSDu

MSDp, MSB MSE F-Prob df HeteroscedastictTestindicates no significant differences 0.10142 0.10402 0.06971 0.01514 0.07551 1,86 Treatments vs D-Control Dose-Response Plot 0.9 0.8 0.7

'*0.6 S0.5 a 0.4 0.3 0.2 0.1 0-1-tail, 0.05 level of significance CD 0

o 01 N

~

CD E

File: sqnl01..081 ] 15data~xlsx Entered by:. J, Sumner Reviewed by:.-4 Page 35 of. 95

'-*"*'*Page 5 of 6 Species: Punenhales Dromelas Client: TVA / Sequovah Nuclear Plant. Outfall 101. NON-treated Daily Chemistry:

Date: 08-11-15 Day I (Analyst identified for each day, performed oH, D.0. and conductvity measurements only.)

.Analyst I

2 H

Concentration

[Parameter pH (S.U.)

DO (ma/L Conductivity (umhos/cm)

CONTROL NON-treated 5--

Alkalinity (nrn CaCO*IL)

U-

Hardness (mc CaCO~iL)

Temperature (0C) i o H (S.U) 7~.*

7.

7 LS*.c DO (ma./L) 10.7%

Conductivity tw(nhos/cma)

Temperature (0C)

IoH (S.U.)

DO (ma/L) 21.4%

Conductivity (umhos/cm)

[ Temnperature (CC)

'U

'U pH (S.U.)

DO (rmgIL) 42.8%

Conductivity (trahos/cmn)

'*Temnerature (0C) fpH (S.U.)

856 DonducivityC 86%2ot*

__________*Temperature

(°C)

,,S*O pH (s.u.)

Ig

[

DO (ma/L) 100%

(m CaCO,/L)

Hardness_

Ti chlorine (ma!!L)

[

O. lbU

Temperature (°C) h-LpW_(s4.)

tDO (ma/Ut 100%

Intake C*Alkalinity

Hardness

]

IT chlorine (maglL) ]

',l.

ic7E

-I~jE Initiatue(C~

.l If FinalS Inb4iti II Final H InWita II Fia

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

AIlsnity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet

. ~~Total residual chlorine was performed on non-treated Outfall 1 01 and intake samples.,

Page 36 of 95 sop AT20 - Exhibit AT20.3, revision 11-01-14

L

-***Pag 6 of 6 Species: Phmephales promelas

... Client: TVA I Sejuoyah Nuler lat0ufai 01NONtreated I

(Analyst identified for each day, performied' De:08-11-15 Day pH, D.O.and conductivity maeasurements onlyj)

Analyst Concent-IParameter tration' pH (S.U.)

DO mnd~.)

Conductivity Aii!!ihoslcm)

CONTROL NON-treated

Alkalnnity A&CaCO*fL)

A~gCaCO*/L)

-u

-~~-

Temnerature (U 0 )

pHa (s.u.)

DO (mn/L3

-le.-7%--

11

~

~-'

-~

I Ii~.~i

~-' '~'

- Temperature (U 0 )

.-l.",S LLI.R p

o -

DH(.u.*

"7_:,7*

.0 *

'~. ~r 21.4%"

I DO(mgIL)

Conductivity I (limbos/cm)

I[

I *Temnerntnre (Ufl I[ I 4

iH (S.U.)

il I lit. -.ff 42.8%

Conductivity

_(tmuniosleni)'

IZ~4L Ia 7

7,

7A.-1.,.'W

Temoerature (UC)

I

--- F "l

[I

[I pH (S.U.)

DO (m' /fL 85.6%

Codctvt I

oo

Tremneratnre (Uc)

'l3.i~ II

-4 pH (s.u.)

1I3."'2J LDo (mg/)

Conductivity 100%

TR en oihos/ (mg)

L "2 'a 111.

DR (S.U.)

[D.(mg/L) 100%

Intake Conductivity

TR chlorine (mag/L) nitial Fnl Initial Final Initial Finnaliia R.w.

i

,.y*eperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table locatedA on Page 1.

[ ' 57*]fflal nity, hardness and total residual chlorine perflormed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet

k [*~y*.Total residual chlorine was performned on non-treated Outfall 101 and Intake samples.

Pa~i 37 of 95

.SOP AT20 -Exhibit AT20.3, revision 11-01-14

a.

S Envlm'onmental Testing Sokattans, Inc, TVA I Sequnyab Nuclear Plant, Outfall 101 - Non-treated August 11-18, 2015 Pimephalespromelas Chronic Whole Effluent Toxicity Test

EPA-821-R-02-013, Method 100040 Daily Chemical Analyses Project nqmber:.

10842 Con1centratlon Param~eter Dm[i Dm2 m3 D4 8DaS Dm p______

(SU)

Iiil Fna nta Fia Iial Fnl ntal Fal Initial Final Initia Final Inia Fnl H (fl7.7 762 7.6 760 7.7 739 7.9 742 7.68 7.48 7.711 7.6

.5 74 DO (mnlL) 78 79 79 80 78 70

.6 79 7.8 8.0

7.

77 Conductivity (Omnbos/emn) 32 0735 1

326 7,327 318.327L Control Alkalinity (agI CaCOO) 59 6

61 Hardness (mgfL CaCO,)

86*

4

~

.88 Temperature *C)2.

46

2.

46 47

2.

48

2.

24.8 24.6 24.81 244 2.8 44 pH (811) 7"7 76

.4 75 77

.5 77

.9 7.71 7.48 7.78!

7.6

.9 73 DO (nmalL) 78 79 79 80 79 69 77 79 7.8 8.0 7.9 7.78,.

10.7%

Conductivity (jimbos/em) 3(

(

0 9

311 291 302 r____

emperature (0C)

2.

244 28 243 48 2.6 49 2.5 24.8 24.5 24.81

24.

248 42

_H (SU) 7.8 75" 75 133 75 72 7.70 7.46

'7718 26 277.3 214

_DO (mnL 78 80 79 80 79 69.8.0 7.8 8.0 8.0 78

7.

292 1od 283y(pnhscm.

2880301

-t 280 2[q*

28

_Ternperatur ICO 50

2.

48

2.

49

2.

49

2.

24.8 24.5 24.8

24.

248 44 pH (81)76 76

.8 7.1 77..2 7

3 7.70 7.46 7.80 7.65 777 74 428%

OfinmL) 79 80 80 80 79

.0 781 7.8 8 0 8.0o.8 78 4,%

Coaduytivi cwpm 26v

.5

/ebm) 263 256'

_______Temperature

('C) 50

2.

24.8 24.5 24.91 24.4 29 25

2.

4 49

2.

49 2.

pH(SU) 76 7.0 77 7.1 77 7.3 70 36 7.67.

7.36 7.79[

7.68 777 74 DO (mig/L)

7.

9 81 7.9 8.0 8.1.

7.

78 85.6%

ConductivIty (gtmhos/em) 20 0

z 0

0 0

209 t'*

209 09 jI,

_______Ternperature

C

2.

45

2.

45

2.

44 29 2

24.8 24.7 24.91

24.

49 2.

pa (SU')

76 7.0 77 7.2 77 7.0 77 737 7.67 7.42 7.'80!

7.6 7.8

.4 8.D81

.0 8.m79

.5

7.

81 7.9 8.0 8....

7.8 Conductivity (gmhos/ern) 19

.8686811921819 100%

Alklnity (mgIL CaCOs) 6967.73..

Hardnes (zng/, CaCO3) 767272 Total Residual Cholorie (malL)

<01

<.0<0.10

_______Temperature (oC) 251 2..48

2.

49

2.

50

2.

24.8 243*

24.91

2.

4.

4 pn (S11) 7.9 76

.9 75 77

.1 77

.6 7.67 7.53 7.7 7.7'779."

DO (mgIL) 80

.1 80 81 80 69 79.1 7.9 8.0]

81 7.8 7.87"

Conductivity ('mnhos/cmn) 189 18 8

8 191 '.::+.i'.

191

++'

191 100% Intake Ailcallnity ng/L CaCO,)

65.69

-73 trness (migILCaCO%)

7272--72 Total Residual Chlorine (malL)

<01

<0.1

<0.10

~t~Yih

,______Temperature

('C)

2.

245 29 2.4 49 2.5 50 2.6 24.9 24.6

24.

245 2.7 2.

File: sqni 0l_08ili5chem.xtsx Entered by: J. SItiner Reviewed by:

'""*'*°*"Page 1 of 7 Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1002.0)

Species: Ceriodavhnia dubia Client: Tennessee Valley Authority Facility: Seauoyah Nuclear Plant NPDES #: TN0026450 Project #:

i *'.

County: Hamilton Outfall: 10._1 IDilution preparation information:...

.Comments:

Dilution prep (%)

I10.7 I 21.4 42.8 I 85.6 I 100 Efiluent volume (mL) 267.5 535 1070 2140.2500 Diluent volume (miL) j2232.5 1965 I 1430 I 360 I. 0 Total volume (mL) 2500 2500 2500.

2500 2500 7 "o -- "..

.*F Test orpanLsmn sourc~e information:

Test information:

1.

i&---.............-

Ol'Ranism age:

' 2.-110o1r5 Old Kendonuzng template color:

between:

Date and times organisms were born O4e%O *

(

Incubator number and shelf Culture board:

. location Replicate number:

11213 l 4i5161[711 91 10 Culture board cup number: "*I*It

' 'L.,[,b~.L1..

"1 YWT batch:

~*t Transler vessel information:

. H * "* 14. S.U.. Ternperature = 3t.

'C Slnsrmbth

%g[

Averae transfer volume (nmL:

0.0 121,,mL

,,S.......

atc

..x 7 "-

t

¢..

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

08-1-15

__________OtAl

,$oli.o\\

IOL 2

08-13-15

,-1____ I~o *".1 10 I' t.*L 4 08-14-15 Og[ "/.

0.10-i$

I$@*tl2.O4

~o t~l, d 5

08-615-15

~

a:t.,o-t$. tSo*,'I.ot 1%eeo.0 k/

6 08-17-15 Ot4fr~~t~

cl~t 7

08-18-15

.. 1 Control information:

Summary of test endpoints:

SControl-i Control-2 Acceptance criteria

% of Male Adults':....

(57.

,7..20%

7-day LCs5 9

7

% Adults having 3'dBroods:

itl1 bb'

IOi,

>.80%

NOEC

% Mortality:..

C7.

- 20%

LOEC

">IO)jl, Mean Offspring;/Fernale:

.\\

"Ž**

>15.0 offspring/female ChIV

Page 39 of 95 SOP ATI 1 - Exhibit AT1 1.2, revision 11-01-14

m**.***=;kPage 2 of 7 Species: Ceriodaphnia dubia Client:.TVA/ISeau~oyah Nuclear Plant,.Outfall1101 CONTROL-1...

Surv, Date: 08-11-15 ivai and Reproduction Data Replicate number

Day, 1

2

....3.....4.

5

.16

,7

,"'8...

9 10

. 1

[_Yngrodee c* {*

O C

{

C) j"""

0 1*

)

lAdult mortality L.. k--

L...

-.. LI I____

g__

2/ *yogprodneed O4 f o

]

jO 4o I

dut orait-

_4_[ Youngproduced A.....

ŽL[I"

' **LJ'** "1'%

I~ dn m__i

'! I

'*- I l

. jl *1~ >-l* '

Adult mortality I_

-1__'-

6 Young produced A

tl Ll O

0 I*.1*

I i

-c 0

"I" I--

C'

-~\\

C-'I Adult mortality.

IITotal young produced I 'A* I*. *-

I

'o SFinal Adult Mortality 1+/

I. '-

g X for 3wi Broods ZI.

Z Z*

N~ote: Adalt moartedlt (L -lOva.D-dead), Sn -split broad (atagl broad spilt bewen two days). CO - cny over (offsprta aredove wt et dardngtra e).

[ Concenatratlion:[

7

[% Mortality

[Mean Ofs tin male:

I CONc: 10.7%

Survival and Reproduction Data Day___

1 2

3 4

5 6 17T8 9

i0 I Autmraty I....

X_.-

1*

zt.i. L.{t.

2it Young produced

.f O...)

C b il*

l Adultnmortality L

J]......

5 [ Young produced 1..

SIAdnult ortality. F T[

L" 6 Youn prdue /1 jk.

4%-."*-%

'j JL--

O 11 I

Adult mortality

~

L 1i- ~.J-._~__

Yovung produced

~t*

J% t* It if i*

I' Iq 1% W Total young produced

}

!1 J

si, si

o Final Adult Mortality L.

W L

L L.

wans o

te:Aot mnortaiy tb a Ive, U = 000aa), omn

= spar aroota(sngle arosol spait asaweos two says). LU -- C '",over (otsapraa earnedl over w~fr a~lts duratrausnater)

Concentration:

% Mortality:0' Mean Offspring/Female:

I *o' I

% Reduction from Control-i:

'3o'*

Sad.paad.nr R.a5.wb~

Page 40 of 95 SOP AT11I - Exhibit AT1 1.2, revision 11-01-14

ia~aPage 3 of 7 Species:. Ceriodaphinia dubia Client: TV'A / Sequoyah Nuclear P~l.ant. Outfall 101' Date: 08-11-15, CONC: 21.4%

Survrval and.Reproduction Data*

i:

i" i:

1 Replicate number'

Day, 1

2 3

4 5

6

-7[

8 9110 1.[Young prodnced "j fl

)

C

)

JflilC)

(*

C)

C)IC SAduit mortality 1

17 1

...1*

2 Young produced

].*

[

)

I *.I

. 1( i 3 I Youngprodcdn*

r 3J0 I

IO 0

O Adult mortality

'---I L.--

I-'1' 4

[Young produced

~.

,I TT

_[j-I1 Adl oraiy _ j j'-

1.II'I-I.....

6 Young produced C) 0 03 0

C 0*

I',

l

..ot'l h'

i J-I.

Total youngproduced I '* I

. I Final Adult Mortality

%--... *., t I

Note: Adult morlaliy (1. = live, D = dead), Su =split brod (struir brood split between tWO days), CO jr *o~ver (offepinn; carded over with adult dumagtronster).

IConcentration:

%M*ortality:

SMean Offspring(Female:

% Reduction from Control-i:

.Lu**

CONC: 42.8%

Sur, ival and Reproduction Data 1

Young produced 0

Adult mortality

~

3 Young produced J

Adult mortality F

0

~ ~

Fnl Adult mortality J

~L 5

Yot:A u

ngt prort L=*w*D=dueac*S bod (sigl brood pLJ t 1'LIee VtwdysO

'~ ove (ofping n Ied ovrwt dl 6dgtes Concentration:

% Mortality:

7,.

SMean Offspring/Female:.

']

i. _

% Reduction from Control-1" SOP ATIi] -Exhibit ATl 1.2, revision 1 1-01-14

'age 41 of 95

Page 4 of 7 Species: Ceriodaphnia dubia Client: TVA,,/Sequoyah Nuclear Plant. Outfall 101 CONe: 85.6%

Survival and Reproduction Data Date: 08-11-15

_ocYoung producedf....j..

j

-Ii SAdult mortality

.J

]

I' I

-I I

__L.

2 1_1 Young produced..

{*

dl otlt

,.X "

)~

L C

JAdult mortality 4..4 3Young produced 1II1fC

~

Adult mortality

-. I

-.-- I I I IAdult mortality

I"

%- ].

j...

[.

f t

[ *

[___

6 Young produced hi *10 I O I t31C*

01l0 v

,r I

4-I Adult mortality

+.

J. __

7 Youngproduced I

%~I i"' ~ ~

1'5 I I 1~l V~I I~ I Final Adult Mortality IL4*

.j

..(

I

%-I~

1 Note: Adalt malrality (L refve.

D dead), SB =split broad (siagl br'cd split betw~eeIwoday), CO=c

-ay ovla~ngurr~antd oar with adultd tngranafor.

Concentration:

Mean Offspring/Fmale:I

% Reduction from Control-I:

CONC: 100%

S urvival and Reproduction Data I

____________Replicate number____

11"...2 3...,4~

6 7

s 9.11o 1

oung produced 0

n'

- 0

[Adultrmortality IL

-W1

",J,*---

[Youpgproduced I ff1C I O) KI

..C I

ab

' - Adult mortality

~

_ Adult mortalityf........

4 YonArducedrt~t Jx-.I...

W-tx--

x-I I x.I '1i*I S-I*

~[ Adult mortality I___

__1_

__[*

1 a-j 7"Young produced 1

t*

t l*

q it:l

'Final Adult Mortality

J 7

1.....

Tf

/,vo*; ItODlX lU*JIlallty Qb -- llVi* L*

Ql*g.I, *)D = Spill DI*OO t SlllglO Ofoon Spln ooP,,vg,011

[wo I:la'J8 h Uu

Gal* : *er *.oxlapml* *)T[*] OVer "*JUl [tOWl OUTfflg (('alSSl*"}.I Conuznnration:"

% Mortality:

0.,

Mean Offspring/Female; I

% Reduction from Control-i:,

7*

IndoOWdoat bsd.o. by VIkyO.t'rn.o.

Page 42 of 95 SOP ATiI - Exhibit ATI 1.2, revision 11-01-14

'° Page 5 of 7 Species: Ceriodaphnia dubia Client: TVA / Sequoyah Nuclear Plant, Outfall 101 CONTROL-2 Survival and Reproduction Data Date: 08-11-15 j___________Replicate number Day 1

2 3

4 5

6 718 9

10 SYoung produced LO f I'

I

<oi 0I..-..*,- "

__Adult mortality K'I*

t'= i -fl*

Z 1 4

Young produced b

b I*'-*I 5

Young produced

$*[*

I*

I *-I1% I*.

I*

l*'*l.*i v-B Adult mortality

= a T

-. 4-..--

-~-

I -i-.---

F -~-,.-----

2-Is L.)I L) 1 1 T2 1 L) 1.J 1 Li I ~...i Adult mortality L_.

L_.

'L__

L..

L I

Iq k..I k_-I

=.'...i

i.

I I

I Total young produced

'2 S

j "Lt a*

.o 0

Xfor 3rM Broods

.j..*

,'*..,_m....

N.ote: Adult siortelity (L f live 1 D deed), SB* split brood (sinole brood split between two daya),

CO - wiscry over (offsprina tu*rned over with adult during tre~ssfhr).

SConcentration:

%Mortali:

I 1.

Me~an Offspring/Eemale" CONe: 100% Intake Survival and Reproduction Data Replicate number_______

Day 112 3 14 5

6 7 T8 9

10 "I'Young produced[

1*

T

-I Adult mortalit s

.4t*

j Adl

-I

__Young produced

.J...Q.0 0 0 0 1 L li Adult mortality J \\...

\\...

~

1

.4 J Young produced

" I

'I1 N 1i*

L t,- s.l*..

"W'L Adult mortalityIk4 W

C

'1--

II Young produced f

5 I 1 5

i i

T I

7[

Y ouogproducedl!,

I r4 to iO I<

O I

I O "10 I~

%)

I d~mmly.O

I

I *-I

-I t-I t-I

Ii LIL Total young produced s

1

8 " *'

,-' 3Aq Final Adult Mortality

..... -- '.--L-1L_

Wwe: Adult mortality (1 llive. D = dead), SB split brood (single brood split betvwia two days). CO = carty~

r (boe r

iaofsn curled over with adult d u,,rin*trnfe I58.p.,d.,t teilewby 1 0.y E. K.uwlI tk~

Page 43 of 95 Concentration:

% Mortality:

)1 SMean Offspring/Female:

I

.% Reduc~tion from Controlo2:

2 IJ.

SOP ATI 1 - Exhibit AT1 1.2, revision 11-01-14

TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated August 11-18, 2015 a* En~mnmenltaI! "rus;~Sol utJoti Inc_

Verification of Ceriodaphnia Reproduction Totals Control-i 24 ACt 0

ton Day 1 ~jjTta1 1

0 010 0

2

-3 0

0~ 010 0TIT~~

0 3I4 5141414 314 33 5

13 11 10 1.2 10 12 12fi13Th1i 11 I114 6

0 0

0

.0 0

0 0 J0j0j0 0

J 7 4 16 15 13 15 14 17 1151171161 152 Total 30 30 279 28 29 30 33 J31J3013113011 10.7%

Dy Replicate number Total Da 3

4 5

6 7

6 9

10 x

oT0 0

0 0

2 0

0 0

0 0' 0 0

0 0.

3 0

0 0

.4 4

4 4

3 3

4 3

3 4

3 35 5

1.2 11 13 12 12 II 12 10 10 1]2 115 6

0 0

0 7

17 15 IS 18 15 16 17 17 17 15 162 Total 33 30

-32 33 3,0 3,1 32 30 31 30 312 21.4%

Re iliate numberTol Day 1

2 3

4 5

6 7i S

9 10 Toa 1

0 0

0

0. 0 0

0 0

0 2

0 0

0

-3 0

0 0

0 0( 0 0

0

.0 4

4 Il3 4

4 5

4 4

4, 3

4 39 5~

1 312 13 10 10 111I2121312 118 6

0 0

0I 0

0 0

0 7

17 18 15 17 1716 1416 1517 162 Total

34. 33 32 31

,32 31 30' 32 31 33 319 42.5%

Day 1

2 3

4 5

6 7

S 9

10 Tol 1

0 0

.0

2.

0 0

0 3

0 0

0

0 0

"6 0

0 0

0.0 7

17 17 19 15 16 15 18 15 15 15

'162 Total 34 33 35

30.

31 32 31 33 31 31 321 DyReplicate number Tota Dy 1

2 3

4 5

6 7

8 9

10

1.

0 0

0 2

0 0

0~

3 0

0 0

4 4

5 5

5 4

4 5

6 4

4 46 5

13 13 22 14 12 12 1.3 13 13 13 128 6

0 0

0 7

18 16, 18 15 19 17 19 15 17 18 172 Total 35 34 3

4.

3 33 37 34 34 35 4

1 0

0 0

7i*

0 0

0 3

0

0.

0 0

.0 0

I0 0

0 0

0 4

6 4

4 5

5 5

4 4

5 4

46 5

12 14 14 13 14 14 12 14 12 13 132 6

0 0

0 7

17 18 17 171I617 19 19 19 18 177 Total 35 36 35 35 35 36 35 37 36 35 355 Control-2 Day 1

2 3

4 5

6 7

8 9

10' oa 1

0 0

0 2

0 0

0

0.

0 0

0" 3

0 0

0

0.

0 0

0 5

11 11 10 12 11 12 1.0 10 13 10 110 6

"0 0

0 0

0 7

16 14 16 13 15 14 16 6 13 14 147 Total 39 23 29 28 30 29_ 30 30 30 27 291 Relicate numberTol 1

0 0

0 2

0 0

0 3

0 0

.0 0

0 0

0 4

5 6

5 4

4 5

5 4

4 6

48 5

12 13 13 14 13 14 14 12 13 11

,129 6

0 0

0 1

17 18 20 18 19 16 19 17 15 17 176 Total 34 37

.38 36 36 35 38 33 32 34 353

File: sqnlDl_0l8111SaataxJsx Entered byr J. Sumner Reviewed by:

A 0

CD Cl'

... Environmental Testing Solutions. inc.

TVA / Sequoyah Nuclear Plant, Outfall 1101 Non -treated August 11-18, 2015 Ceriodaphnkz dubia Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1002.0 Quality Control Verification of Data Entry, Calculations, and Statistical Analyses Projectl numbern 10842 Concentration Repl~iae number Survival Average reproduction Coefficktte Po~nut reduetlee from

(%4) 1 "2

3 4

5 6

7 8

9 10

(¶4)

(ollspriaglfemnale) vartifiCK (%)

carontr (%)

Control - 1 30 30 29 28

'29 30 33 31 30 31 100 30.1 4.6 Not applicable 10.7%

33 30 32 33 30 31 32 30 31 30 100 31.2 3.9

-3.7 21.4%

34 33 32 31 32 31 30 32 31 33 100 31.9 s8

-6.0 42.8%

34 33 35 30 31 32 31 33 31 31 100 32.1 5.0

-6.6 8.5.6%

35 34 35 34 35 33 37 34 34 35 100 34.6 3.1

-15.0 100%

35 36 35 35 35 36 35 37 36 35 100 35.5 2.0

-11.9 Control.- 2 30 28 29 28 30 29 30 30 30 27 100 29.1 3.8 Not applicable 100% Intake 34 37 38 36 36 35 38 33 32 34 100 35.3 5.8

-21.3 Outfall 101:

Dannett's MISD value:

PMSD:

Intake:

Dunnett's MSD value:

PMSD:

11254 4.2 1.280 4.4 MSD =

Minimum Signfcant Difference PMSD =

Percent Minimum Significant Dilterence 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 whale effluent toxicity test.

Lower PMSD bound determined by USEPA (tO0' percentile) =13%.

Upper PMSD bound determined by USEPA (90"' percentile) = 47%.*

Lower and upper PMSD bounds were determnined from the l10th and 90th percentile, respectively, of PMSD data firon EPA's WET Interlaborutory Variability Study (USEPA, 2001a; USEPA, 2001b).

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. Cincinnat, OI. L USEPA. 2001is, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes 1 and 2-Appendix. EPA-821-B-01-004 and EPA-821-B.0lI-005.

US Environmental Protection Agency, Cincinnati, 011 File: sqn101_081lil1data~xlsx Tablepopulated from associated 'Verifcation of Ceuiodaphnia Reproduction Totals' spreadsheet.

Sprenasheet entered by: J. Sumner Reviewed by:.,6 S....

TVA / Sequoyah Nuclear Plant, Outfall 101 Non-treated August 11-18, 2015

Statistical Analyses

'0T 0

0 r1bfmmll~lSul~n Ceriodaohnia Survival and Reproduction Test-Reproduction Start Date:

811112015 Test ID:

CdFRCR Sample ID:

[WA 1SQN, Outfall1101 End Date:

5/18/2015 Lab ID:

ETS-Envir. Testing Soi.

Sample Type:

DMR-Dlschlarge Monitoring Report Sample Date: August2015 Protocol: FWCHR-EPA-821-R-02-013 Test Species:

CD-Ceriodaphinla dubia Comments:

Non-treated Cons-%

1 2

3 4

5 6

7 8

9 10 Control-i 30.000 30.000 29.000 28.000 22.000 300O00 33.000 3.Z,*.

"fl.000 31.000 Control-2 30.000 20.000J 29.000 28.000

=,u.u0 29.000 3u.u00 30.000 30.000 27.9000 10.7 33.000 30.000 32.000 33.000 30,000 31.000 32.000 30.000 31.000 30.000 21.4 34.000 33.000 32.000 31.000 32.000 31.000 30.000 32.000 31.000 33.000 42.8 34.000 33.000 35.000 30.000 31.000 32.000 31.000 33.000 31.000 31.000 85.6 35.000 34.000 35.000 34.000 35.000 33.000 37.000 34.000 34.000 35.000 100 35.000 35.000 35.000 35.000 35,000 38.000 35.000 37.000 36.000 35.000 Intake 34.000 37.000 38.000 38.000 38,000 35.000 38.000 33.000 32.000 34.000 Transform: Untransformed Cons.%

Mean N.Mean Mean Bin Max CV%/

N Control-i 30.100 1.0344 30.100 28.000 33.000 4,553 10 Control-2 29.100 1.0000 29.100 27.000 30.000 3.782 10 10.7 31.200 1.0722 31.200 39,000 33.000 3.940 10 21.4 31.900 1.0982 31.900 30,000 34.000 3.753 10 42.8 32.100 1.1031 32.100 30.000 35.000 4.969 10 85.8 34.800 "1.1890 34.600 33.000 37.000 3.107 10 100 3.5,50 1.2199 35.500 35.000 37.000 1.992 10 Intake 35.300 1.2131 35.300 32.000 38.000 5.829

. 1[0 1-Tailed Isotonic t-Stat Crltical MSD Mean N-Mean 32.687 1.0000

-2.000 2.287 1.254 32.587 1.0000

-3"282 2.287 1.254 32.557 1.0000

-3.647 2.287 1.284 32.557 1.0000

-8.208 2.287 1.284 32.587 1.0000

-9.847 2.287 1.254 32.567 1.0000 Koimogorov D Teat indicates normal distribution (p > 0.01)

Bartiett's Test indicates equal variances (p = 0.34)

The control means are not significantly different (r = 0.09)

Hyvpothesis Test (1-tall, 0.OS)

NOEC LOEC Ci Dunnett's Test 108

>100 Treatments vs Control-I Statistic Critical Skew Kurt 0.90242 1.035 0.57339

-0.005 6.68798 15,0863 1.79828 2.10092 TU MSDu MSDp MSB MSE F-Prob df 1

1.254 0.84166 42.7057 1,5037 5,4E-14 5, 54 rtv Linear Interpolation (200 Resamples)

Point SD 95% CL Skew IC05

>100 IC10

>100 IC15

>100 1.0-IC20

>1000.

1C25

>100 o.6 1040

>100 0.7 iC50

>100 0.6 a0.4-S0.3" 0.1"

-0.1

-0.2

-0.3 "

0 50 100 150 Dose %

Dose-Response Plot 1-tail, 0.05 level of significance R.~4~w by ISU.u E. Keen.,

Page 46 of 95 1diis squlOl 0811 l~dat,.xisx Entered by: 3. Sumner Re'dewed b: e

TVA / Sequoyah Nuclear Plant, Outfall 101 - Intake Non-treated August 11-18, 2915

'Er S

Statistical Analyses Ceriodaphnia Survival and Reproduction Test-Reproduction Start Date:

8/11112015 Test ID:

CdFRCR Sample ID:

TVA/ SON, Outfall 101 - Intake End Date:

818182015 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring R~eport Sample Date: August 2015 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 30.000 29.000

.28.000 29.000 30.000 33.000 31.000 30.000 31.000 Control-2 30.000 28.000 29.000 28.000 30.000 29.000 30,000 30.000 30.000 27.000 10.7 33.000 30.000 32.000 33.000 30.000 31.000 32.000 30.000 31.000 30.000 21.4 34.000 33.000 32.000 31.000 32.000 31.000 30.000 32.000 31.000 33.000 42.8 34.000 33.000 35.0.00 30.000 31.000 32.000 31.000 33.000 31.000 31.000 85.6 35.000 34.000 35.000 34.000 35.000 33.000 37.000 34.000 34.000 35.000 100 35.000 36.000 35.000 35.000 35.000 36.000 35.000 37.000 36.000 35.000 Intake 34.000 37.000 38.000 36.000 36.000 "35.000 38.000-33.000 32.000 34.000

'Transform:

Untransformed 1-Tailed Conc-%

Mean N-Mean Mean Mini Max CV%

N t-stat Critical MSD Control-I 30.100 1.0344 30.100 26.000 33.000 4.553 10 Control-2 29.100 1.0000 29.100 27.000 30.000 3.782

.10 10.7 31.200 1.0722 31.200 30.000 33.000 3.940 10 21.4 31.900 1.0962 31.900 30.000 34.000 3.753 10 42.8 32.100 1.1031 32.100 30.000 35.000 4.969 10 85.6 34.600. 1.1890 34.600 33.000 37.000 3.107 10 100 35.500 1..2199 35.500 38.000 37.000 1.992 10 Intake 35.300 1.2131 35.300 32.000 38.000 5.829 10

-8.403 1.734 1.280 Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01 )

0.95757 0.868

-0.2237

-0.356 F-Test indicates equal variances (p = 0.08) 3.49541 6.541 09 The control means are not significantly different (p = 0.09) 1.79926 2.10092 Hypothesls Test (l-tall, 0.05)

MSDu MSDp MSB MSH.

F-Prob dft Homoscedasti t Test indicates no signlficant differences 1.2795 0.04397 192.2 2.72222 1.2E-07 1, 18 Treatments vs Control-2 Dose-Response Plot 1-tail, 0.05 level of significance bd.p*,d,~t R.~.wI~y elley E. K..,,*,,

Page 47 of 95 File: sqn1 01.08111 5data.xlsx Entered by: J. S~jsnr Reviewed by:

0 Page 6 of 7 Species: Ceriodaohnia dubia Client: TVA /Sepuovyah Nuclear Plant. Outfall 101 Dally Chemistry:

Date: 08-11-is Day

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

0 I

1 I

2 Analvst

_* IIb

/V6 I &

II Nb I TV"_jII *.

-~

U Conceutration j Parameter lI (i S.U1 i127d

-I -~---~--'-----4I-~-~------

DO (malL)

II CONTROL Conductivity j __________

(,nnboslcm)

Afl{~jlinfty (mg CaCO3IL)

Hardness (mg CaCO~lL) 7,70 7.q 307 b\\

Temnerature (¢Cq

1m aH (S.U.)

"*. In*i.

I1 7.7*

O 7;7*

II 7.. &_q - I.....

r DO (mgJL) 10.7%

Conductivity I

GO

Temperature (°C)

l'$1mLm*m.

nH (S.U.)

21.4%

Conductivity

~

(~tnihoslcmn)

~

I~A1 L

Temuerature (*C1 L~. I pH (S.U.)

DO (mgIL) 42.8%

Conductivity

(iimboslcm) 2Aj~

Tewperature (0C) 1.$.0 b'L. i pH (S.U.)

DO (mglL) 85.6%

Conductivity

Tcmperature (0C)

~5L.

,A.

'1 Ab y_&__ 7.7*

_ IJ. M n...*

.0I nHl (S.U.'

-~~.;q 1L7.

JL DOo

,0

"--=

Conductivity t(

Alkalinity1 100%

(mg CaCO3/L) t

Hardness I*!*'I2

TRl chlorine (raglL)

Conductlvity 100%

Alkalinity

]

Intake (rag CaCO3IL)

Hardness

T'R chorine (mg/L)

0.

0 Initia~fif=

=l Finalf*

Initial Final*

Initial Final

Temperatures perforned 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 identilied o~n the bench sheet specific for each analysis and transcribed to this bench sheet by:

J~4.,.hy Ik~

Pagle 48 of 95 SOP AT!I - Exhibit AT1 1.2, revision 11-01-14

Page 7 of 7 Species: Cerlodaphnla dub/a Client: TVA / Seciuovah Nuclear Plant. Outfall 101 Date: 08-11-15 I I ill r

,,L

J',J g

D.

I H

I i

5 Date:

8-11-1 Concen-tration Parameter U pH (S.U.)

Conductivity Oimhoslcm)

CONTROL

Alkalinity (mg CaC0 3/L)

Bardness (mug CaCch/L)

Temnerature I

I i

,---*-.-t i

ii ir" a

-=

pH (S.U.)-

I....

II %7.7 I 7.7D9 II 7.C_.?

I 7-7 DO (rn~/L)

~

II II-II

n m*e I

JJLffta 1 LondIzctisttv-I-(umhoe/emni a

~

Temnerature t°Q)

'., A IM.9 DO(m~/L) 21.4%

Conductivity

.ŽoI

(~mhosIcm) ii oH (S.U3l DO (mn/Li R.~n

~?. I

.74 42.8%

Conductivity

~ -~t.,-,--

-~----

I

[.;

Temperature (°C) a.AA Itt m*..$

LJ

-~)l.

0 iL 9

9 II

~.-

I~ -

I ~

41.~

U iJH (s.U.)

74.&.

,7 ?

7.1.6

J "-*

if' r*

) - Lv*." -*

_- "" W II -- 'e-

Ik------t-z--J-.*---

DO (rig/L) 85.6%

Conductivity (i*nnhos/cm)

A'iLiL 7.1'

ŽL2

Temnerature (°C*

"%I\\% *

",4,r

-L*

-I

-~

U ~

pH (8.U1.1 1,,,7II 7.7Li I

7, e,4 I

r*" L. -

It" II

-JT, DO (regA) 100%

SConductivity

Alkalinity (rag CaCO3/L)

Hardness

TR chlorine (mag/I) oH (S.U.',

p Final/

Initial Final DO (ma/LI 100%

Intake

[,oq I

Alkalinity

Hardness

TR chlorine (rag/L)

Temperature*C*_

.0 _

L J

RoC4,,~

b~

jc.IIov L ~

Pag Initial II

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

, AC,*$ ;

SOP1 ATI 1-Exhibit AT1 1.2, revision 11-01-14

=It TVA / Sequoyah Nuclear Plant, Outfall 101 - Non-treated August 11-18, 2015 Ceriodaphuic duhia Chronic Wihole Effluent Toxicity Test EPA-821-R-02-013, Method 1002.0 Daily Chemical Analyses Project number.

10842 Concentration Parameter Ba0P

]

Pa 3 Pa D

D a

pH_____

(SU)__________

nta Fia Initial-Final nial Fal Initial Final Initial Fia Intl Fnl Iiil Fnl 9 H(I)77

.4 7.63 7.65 7.0 76

.98 7.63 7.68 77

.1 75

.5 77 DO (nig/L) 7.78

7.

8.0

7.

76 7.6

8.

7.8

8.

79 79 77 77 Cnrl Conduzctivity jjjiuumhos 32 307 30 317 32632 18'Zi&7

_______Temperature (C) 24.249 48 25.1 24.8

24.

24.8 48

2.

52 248

2.

49 2.

InH (SIU) 7.671 7.761 7.741 7.681 7.731 7.681 7.741 7.6 21 7.711 7.721 7.781 7.63 7.19 I 7.72 10.7%

[Conductivity (jzmobskmr)

Ij 307-W 300 1 301 294)

3'1- '

.2911nŽ....

302j *Ž-iit

[Temnerature too' 24.71 25.11 24.81 24.81 24.91 25.11 24.91 24.91 24.9 25.01 24.71 24.8 24.91 25.0 1 *IH (SU) 3 7.68 7.76 7781 7.69.

7.73 7.67[

7.75 76q3 7.70 7.731 7.78[

7.64 7.771 7.71 214 IOm/L 7.8[

7.8 79 8.0...

7.9L_

7.6 7.8 80]

78 81 80o 7.9 7.8l 7.7 Lfem=eratnreCc J

24.7 21_*!

2491 24.8 29 2

.1 2.9.

4.4 49 24 4

50 2.

nfl (QYTh 7 681 7 RDI 7

gl 7 7N 7 7] I 7_65 I "7731 76 31 7_701 7731 7.80t 7.64 777 7_71 20 ~"'~'~

i 80 8.0

'7:9

7. 7J 78 7.81 8.ij tO

_______I 'A

.....I

..- I

.4-, r~~i

~

'.- I 4L.8%

r[nudiiptfrhitv fmmha~clomul 264 rin h **;'

l----I".......

'1 -(

I

-I I

" I I

remnernenre fCn 24.81 25.01 24.91 24.9 24.91 24.91 24.81 24.8 25.01 24.9 nil (Sin 7.681 7.811 7.7 8!

7.72 7.70[

763 7.67 7.73 7

76 j 7.J 7 71 7,,,

.70 791 80o 79j 8.1 8.11

.1 78 85.6%

flfl (inSt.)

w I-

.vuI u.IfMI lflyll*

Uf'ilflU 4'" *LUR I

Temneratnre gOC' a

I--

I DO (onr/I, I.-

Conductivity (,mhosn/cm) 100%

Alkalinity (mg/I. CaL-tO,)

I I

I Hlardness (mgIL CaCti3)

Total,*Reiul,* Chlorine (ma./I) 25.01 25.11

24.

251 24.9 2.

24.9 24.91 24.8

. I I

I 7.67

"~l~**

H

~'J~"'

j

~

I

.1 I

"1*

HA C

.n Tt~mntrahurs (

0t"*

OH (SUfl DO (mall.,)

u * %--el,--,"

l le'vna,el m i m Ih*.-; 4'*t,- dl',a m h *IBm%

100% Intake

,.uflUW*443.svaz.now~t~nmI L

Alkaflnty (mgtL CaCOi3 )

FHardness (mtg/L CaCOi3) j Total Residual Chlorine (ma/fL) 7.71 7.44 7.67 7.80!

7.771 7.811 7.79I 7.74I 7.9 8.0 7.9 8.1 8.1[

8.0 7.81 8.0 184 i

191 I91 l

191'.

2.

250 25.0 24.9l 25.01 24.91 24.8[

25.1l remoerature CoL-I I

1 r

I I

I II I

I I

" I 1

" I File: sqn101l_8111 5chem.xlax Entered by:. J. Suqiner Reviewed by:

SET$

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

Species: Pimevhales [Womelas Client: Tennessee Valley Authority Facility: Sea uovah Nuclear Plant NPDES #: TN0026450 Project #:

IA)LO'A2-County: H~amilton Outfall: 10_1 SDilution preparation information:

Comm~ents:*

Diluti~on prep (%,)

10.7 21.4 I42.8

" 8,5.6

-l100 Each concentration was UV-treated

.Effluent volume CrnL) 267.5 535 1070 2140 2500

'for 2 minutes to remove pathogenic1 Diluent vouei(nin.(L) 2232.5 1965 1430

.360 0

Interferences.

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

[.....

Test information:

isg~nisma e:

f*'

L*.\\V* *Qt.O60 Randomizing template: 1Ne~a*t a

1 JW i ° were born between:

________________shelf location:_____

Organism source:

AToX Batch Pp:" *iO"*

Aremia CHM number: JCHM836 Drying information for weight

________________determination:

a Transfer bowl information:

pH =

"%.'a*

S.U.

Date / Time in oven:

I________t

_________Temperatizr~e

-. "gA.

°C Initial oven temperature:

la "C

'C,@,*

Average transfer volume:

0.1749 niL Date / Ti*m~e out of oven:.I.¢ lV Final oven tempjerature:

_____________________Total drying time:

7tg1gt 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 Analy'st Outfall 101

]

Intake 2

08-13-15 wr A

AO9

. 1

)b4f

)

L

?

6 08-17-15 u

tS6* tq.0g

~I08-18-15 1.1*

Control inorination:

Acceptance criteria Summary of test endoints:

% Mortality:

)*.<20%

7-day LCso

_______o Average weightper initial larvae:

.1%*L NOEC Average weight per surviving larvae:

0.YL,.

> o.25mg/larvae.

LOEC

>1!001.

ChIV

> lIc0i.

IC2 s

%) tt~t~7 j i L

51 of 95 51 of95SOP AT20 - Exhibit AT20.3, revision 11-01-14

Page 2 of 6 Species: Pimephales Dromnelas Client: TVA / Seauovah Nuclear Plant. Outfall 101,

_1V.-treated Date: 08-11-15 Surv'ival and Growth Data Day CONTROL 10.7%

21.4%

ai A

B Cl D E F

,G H

I J

K L

0 10 10 10 I 10 10 10 10 10 10 10 10 10 It )oI t '0i 10 IOl 1r t t*

to tO tO lO

~ IU 10t0 10 It) 1*

(010O10 t0

'0 (0

10

'U.

l0 (0

'U

'0 s

I*

rb

,t

,o0 to l(

Io EQ 1 0 10 t

tO 0

6 I01tO D1t0,O 1(O(tO 0,01 O.O 7I t)

I) 10

'Oit JO EQ1 0

10 tO 10 tO.1)

Date:

o qA

-.l tp122.-\\

3.'

Z,..,~*l

'I~

Analyst: s Weight per initial number of larvae (mg)

= C I Initial number of larvae Hand calculated.

Analyst:~

(E7 0%

1%

"I

'p p

1%

V N

0%

'S

'V.

Average weight per Percent reduction initial number of from control (%)~ 0,'1V"L-i -

a -

p a a

s.i~a1.

c6.-v%\\

- "*..'*7.

I h ~

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

Comments:

Page 52 of 95 Page52 o 95SOP AT20 - Exhibit AT20.3, revision 11-01-14

--*,*Page 3 of 6 Species: Pinephales prom elas Client: TVA / Sewwoyah Nuclear Plant. Outfall1101, UV-treated Date: 98-11-15 Survival and Growth Data Day 42.8%

-85.6%

100%

M N

0 P.

Q

.R S

T U

V W

X 010 10 10 10 10 10 10 10 10 10 10 10 it)

LIt) iIO 10,/)i 1Q. 0 (010 10 2

it o

tO tO lo 0 10(1)t' tOl tO-3____

(0 OID'0 IO O,01l01/

Of0'0

' 0 I

A 1

4 4

4~

~

1 I

l-----t.+/-...

I-~-*

10 (0

10 10

/0 IL)

'U~ '~

(0

/O lO iO I0

/0 iO) lO I.J s___

__0 IQ 11)O tO t

II 0 i 0 /

O

/Q/0 t

'0 0/

6_________(

lO IO D 11)

¢0

/0 I)

/1) (0 10 10 lO 71jc

'1 IQ3

'0 tO 10 so io i

,0 t0

/0 A =Pan weight (mg)

Drate color ode:-

B = Pan + Larvae weight (mag)

Date:

[q t:*-

lj.SL~

tLJ

~.I2qS

~5ZO~Z.2 2.~

M~

C = Larvae weight (rag) -- B -A Analyst:

Weight per initial number of larvae (mg)

C C/rnitial number of larvae HAndalcuated 1 iL...*

Average weight per.

Percent reduction initial number of.

from control (%)

0 1."*'

7o 0.l"Iog[

iO1o*7 0."1t.

- '.A7 larvae (mg)

.o...oes...la,.

ed g bgs klem=misns scS nuulysal Ig =unusually large, d&r =decanted and returned, w -- wounded.

SCommentIs:

SOP AT20 - Exhibit AT20.3, revision 11-01-14

~

Page 4of 6 Species: Phrney~hales promelas, Client: TVA / Sequoyah Nuclear Plant Outffall 101,

,UV-treated Date: 08-11-15 Day 100% Intake 1 0 1 0 10 I10 I

0 l0 IQt lb t: 1> 10 Io S4

, t

'V I0 I O (U

6 lL*[ 10 10 [/0 A =f Pan weight (mg)

Tray color e:: cf_11rC2e n*

r~a Date:

B =Pan +Larvqle, weigbt(rag)

Date:

Ljj.......

C =Larvae weight (rmg) = B - A Band calculated.

1q I

t 0

.~q 6.[

Analyst:

Weight per initial number of larvae (rag)"

= C /Initial number of larvae

$**[

01/

Hfind calculated.

.4_

P

,% ID Analys:

o' I*

Average weight per Perecent redaction iniianme f

from control (%)

6 * **

,, *1*.

larvae (mg)_______

Co~rm.ent 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.

C o m m...............

e SOP AT20 - Exhibit AT20.3, revision 11-01-14

SEnvironmental Testing Solutions, Inc.

TVA/ISequoyah Nuclear Plant, Outfall 101 TV-treated August 11-18, 2015 Pimephalesprometos Chronic Whole Effluent Toxicity Test EPA-821-R-02-0 13, Method 1000.0 Quality Control Verification of Data Entry, Calculations, andt Statistical Analyses Protect anumh~m 10842 Ceu~qieUA latiza~beorF~dlmb~~e*

,PuwfFhtml0B~mt-]auaeI,=v________

Nd forr Coinp1

~

hl' efqmofsdollWe~e stensosl nesr-ri al Lngtllb o.r~qel~eQ Ptcarno rC taJva

,reb (e*,g)(

--A-B umer~roitce e(eoe)

Surnlei~tgltebeer O

t Mm

'a~'eo of urine(tiq

(%)

Itnmb,, Ot ttrroe rnlto. lO-,toto

,stts! (51)

A 10 10 14.13 21.98 7.85 0.785 0.785 onrl

.B 10 10 16.14 22.73 6.59 0.659 0.712 9.1 0.635 (2trl 10 10 15.85 72.40 6.55 0.655 0.659

0.

0.71.2 9.1 Not applicable D

.10 10 14.95 22.42 7.47 0.747 0.747 E

10

,10 15.59 22.07 6.48 0.648

.4 107%_____0_

10 16.75 22.63 S.88

.580.672 9.9 1.810D.0 0.672 9.9 5.6 1.%

G 1 0 10 16.41 "23.67 7.26 0.726 0,2

_______O 10 15.5' 72.77 7.24 0.724 0.724 I

10 10 16.43 23.55 7.12 0.712 0.712 21,4%

1" 10 10 16.37 24,.04 7.67 0.767 0.3190.767 t0.i1.9-7 K

10 10 16.57 22.76 6.19 0.619 6.619 L

10 10 16.36 24.60 0.24 0.824 0.824 61 10 10 15.04 22.59 6.55

'0.655 0.655 4.%

N 10 t0 15.27 22.65 7.38 0.738

.73"6.3 0.738

0.

.2 6.3 1 0

10 10 15.05 23.32 '

7.47 0.747 0.747 P

10 10 15.97 23.50 7.5.3 0.753 0.753

.....Q...

10 10 15.97 24.08 8.11 0.811 0.811 8.%

R 10 10 13.84 21.07 8.03 0.80

.8B.

.03 n

.0 078 61-1.

S 10 10 16.335 24.50 8.15

0.

0815 07 610.8l5 0-0,86.-0.

,, I 10 10 14.40 21.55 7.15 0.715 0.715 U

10 50 13.61 20.98 7.37 0.737 0.737 100%

V.

10 10 15.09 22.22 7.13 0.713 0.782 9.1 0.7 13 100.0 0.782 9.1

-9.9 W

10 10

,16.40 23.09 8.69 0.869 0.869 X

10

,10 15.94 24.05 8.039.

0.809 0.809 Y

10 10 16.607 23.01 6.94 0.694 0,694 100%TInt~ale Z

10 10 16.40 23.63 7/.43 0.743 0.659 4.4 0.743 130,.0 0.699 4.4 L8 Ak 10 10 15.00 21.89 6.89 0.689' 0.689 BB________

D 10 10 15.93 22.63._

"6.70 0.670 0.670 Dunnest's M8SD v'alise Danaeti's MSD valuer PMdSD; 0.1114 15.7 0.0699 MOD =

Mininnun $ignifitont Difrnce PMSD =

Percent Mianimue Significant Difference PMSI) is a meenure oft'est tnncisiae. The PMSD is the mmemeun percent difference between the control and treatment that can be declared ataliticnily sisgnificant in o whole eftlent toxicity test.

Lower PMSD bosmd deternuined by UflEFA (Iothpercentile) = 1.2%.

Upper PMSD botrnd determined lsy USEPA (90th pernt'o.nile) = 30%.

Lower end tupper PMSD bmmds were drctnrisnd from the I10th and 90th percentile, respcetivey, of PMSD data from *EPA'S WET lntedlaboratory Vardabhity Stdy (SEPA.,

2003la; USEPA, 2001bh).

USEPA. 2001a, 2001b. P'mat Reportlo Iterlaboratosy Vex~abiht~y Study of EPA Short-tent, Chronic and Acute Whl~oe Efiluent TosicityTest Methords, Vohones l and 2-Appendix. E1'A-821-n-01-004 end EPA-821-nB-01-o0. US Environmenztal Protect*ion Agency, Cioharai, OH.

File: sq~n10 O 111.5llfdala-uv.glax Entered by:. J. Suppe Reviewed by:

.TVA / Sequoyah Nuclear Plant, Outfall1101 U V-treated E T$

August 11-18, 2015

~Statistical Analyses

nlonm.ntu Teh1ag ohumonl,Inr.

Larval Fish Growth and Survival Test-7 Day Growth Start fiste":

8 "~'z;*:;;

Test ID:

PpFRCR Sample ID:

"WA / SQN, Outfall 101 End Date:

ri/i2j;*2*.

Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Disch~arge Monitoring Report Sample Date: Aid-g-i*r2"15 Protocol: FWNCHR-EPA-821-R-02-01 3 Test Species:

PP-Pimephaies prmmolas Comments:

UV-treated Cone-%

1 2

3 4

0-Control 0.7850 0.6590 0.6550 0.7470

10.7 0.6480 0.5880 0.7260 0.7/240 21.4 0.7120 0.7670 0.8190 0.8240 42.8.

0.86550 0.7380 0.7470 0.7530 85.8 0.8110 "0.8030 0.8150 0.7150 100 0.7370 0.71 30 0.8690 0.8090 Intake 0.5940 0.7430 0.8890 0.6700 Transform: Untransformed 1-Tailed Isotonic Cone-%/

Mean N-Mean Mean Min Max CV%

N t-Stat Criticel MSD Mean. N-Mean fl-Control 0.7115 1.0000 0.7115 0.8550 0,7850 9.113 4

0.7341 1.0000 10.7 0.8715 0.9438.

0.6715 0.5880 0.7260 9.897 4

0.885 2.410 0.1114 0.7341 1.0000 21.4 0.7305 1.0267 0.7305 0.6190 0.8240 11.947 4

-0.411 2.410 0.1114 0.7341 1.0000 42.8 0.7233 "1.0165 0.7233 0.6550 0.7630 6.349 4

-0.254 2.410 0.1114 0.7341 1.0000 85.6 0.7860 1.1047 0.7860 0.7150 0.8150 6.055 4

-1.612 2.410 0.1114 0.7341 1.0000 100 0.7820 1.0991 0.7820 0.7130 0.8690 9.068 4

-1.525 2.410 0.1114 0.7341 1.0000 Ijitaire 0.6990 0.9824 0.8990 0.6700 0.7430 4.449 4

Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wiills Test indicates normal distribution (p >O.01) 0.94413 0.884

-0.2521

-1.0075 Bartlett's Test indicates a ual variances (p= 0.91*

1.5502 15.0883 Hy/pothesis Test (1-tail, 0.05)

NOEC LOEC ChiV TU MSDu MSDp MSS MSE F-Prob df Dunnett'sTest 100

>100 1

0.1114 0,15657 0.00784 0.00427 0.18624 5, 16 Treatments vs D-Control Linoar Interpolation (200 Resamples)

Point S'D 95% CL(Exp)

Skew 0C05

>100 IC10

>100 IC15

>100 1.0 IC20

>100 0.8 1C25

> 100 o.a I040

>100 I050

>ioo 0.7 0.8 0,

0.4 S0.3 re0.2, 0.1, 0.0

-0.1

-0.2 0

50 100 150 DOSe %

Dose-Response Plot

O 06 T1-tail, 0.05 level of significance 0.5 0 0.4 0.2 0.1 File: sqnIl 01_1 llSdits-uv.,dsc Entered by: J. Sumner Recviewed by:

Page 56 of 95

TVA / Sequoyah Nuclear Plant, Outfall 101 - Intake U V-treated August 11-18, 2015 Envronmental Testing Solutions. Inc."

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

_'-i:*.

Test ID:

PpFRCR SamplelID:

"rVAISQN, Outfalll101 -Intake End Dae:

1j Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

DMR-Discharge Monitoring Report Sample Date: A'i~i~t2;*"5 Protocol: FVVCHR-EPA-821 -R--02-013 Test Species:

PP-Pimephaies promelas Comments:

Nan-treated Conc-%

1.

2 3

4 0-Control 0.78560 0.6590 0.6550 0.7470 10.7 0.6480 0.5880 0.7260 0.7240

21.4 0.7120 0.7670 0.6190 0.8240 42.8 0.6550 0.7380 0.7470 0.7530 85.6 0.8110 0.8030 0.8150 0.7150 100 0.7370 0.71 30 0.8690 0.8090 Intake 0.6940 0.7430 0.6890 0.6700 Transform: Untransformed 1-Tailed Cone-%

Mean N-Mean Mean Min Max CV%

N t-statr Critical MSD 0-Control 0.7115 1.0000 0.7115 0.6550 0.7850 9.113 4

10.7 0.6715 0.9438 0.6715 0.5880 0.7260 9.897 4

21.4 0.7305 1.0267 0.7305 0.6190 0.8240 11.947 4

42.8 0.7233 1.0165 0.7233 0.6550 0.7530 6.349 4

65.6 0.7860 1.1047 0.7860 0.71 50 0.8150 6.055 4

100 0.7820 1.0991 0.7820 0.71 30 0.8690 9.068 4

Intake 0.6990 0.9824 0.6990 0.6700 0.7430 4.449 4

0.348 1.943 0.0698 Auxiliary Tests Statistic critical Skew Kurt Shapiro-Wiik's Test indicates normai distributio~n (p > 0.01 )

0.9396 0.749 0.32208 -1.1983 F-Test indicates equal variances (p = 0.26) 4.34562 47.4683 Hy.,othesis Test (1-tail, 0.05)

MSDu MSDp MSB MSE F-Prob df Homoscedastic tTest indicates no significant differences 0.06987 0.0982 0.00031 0.00259 0.73997 1, 6 Treatments vs 0-Control Dose-Response Plot I

0.9 0.8

'0.7

0.6

0.5 o 0.4 0.3 0.2 0.1 1-tail, 0.05 level of significance bd.p~8oot

~

~v Page 57 of 95 File: sqnl 01..08115data-uv.xlsx Entered'by: J. Sumner Reviewed by:.

~

Page 5 of 6 Species:.Pimehales vromelas Client: TVA / Sequoyah Nuclear Plant. OutfalI 101. UV-treated Daily Chemistry:

Date: 08-11-15

'I(Analyst identified for each day. v*fre *,DO and conductvity measurements only.)

Analyst Concentration

~H (SU.)

DO (ma/

~

Conductivity (umhos/cm'*

CONTROL UV-treated 4

S*Ha-rd ness S*Tempe~rature (0C) km S 10.7%

nil (S.U.'I conaoeuvny

~

A, I 'W~c-1~i~77t

Temperature (0C)

"t.H..

I ' "L*, O "ii.,'.

"tH.*t A

~I h ~

(1,t1 7, (a#

17,.,q" 7,'.')

211.4%

DO (mg/b)

IiiI~IZI1i 7

I 7..9 ii

,,~

Conductivity

~

I

- ~'Temperature ~'C) li.L *

'LL.1 i'"16.0 LVo

-IS.0 S

U I

-~..

clH (s.u.*

~----~-----

-=-=-

0 I

I -~*=--------1I---

DO (rag/L) 1-is 42.8%

Conductivity

'Terneperatnre

)

85.6%

Conductivity 2~

Conductivity 100

Alialinty L

Hardnes

TR chlorine (rag/L)

£'.~O=

Conductivity 2

100%

Alkalinity t-

intake (tu C aC. 3/L).*

Hardness S*TR chlorine (rmg/L)

.0-l sal Initial""

Final Inta ia I

I Initial II Fii d*~,peratures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal lnfobrmation table located on P*age 1.

o"-.*hlati ity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet T otal reida chloin wa perorme on n on-rae Outal 10 a, o

nak sao.mples.

age 58 of 95

'sop AT20 - Exhibit AT20.3, revision 1 1-01-14

Page 6 of 6 Species: Pimeyhales pr'omelas Cli. t TVA / Se u yah Nuclear Plant Outfall1101* UV-treated Date" 08-41-15 Day Analyst.

="

Concen-Parameter fratiofl I

Hardnress o',,,~(tg caco3(L)

\\&

Temperature (OQ)

.1Q" t

"I.0

".6 l.'

-O*m

/-)

-_.4.,

,*Temperature (°Op

'*-S'o--]

.0 "A

1.

21.4%

Conduetivity.3 DO (*Tmgeatre°)

T"~0**1

).

" 2 DT horil(mgit )

0

&0

..']*Temperature

("C) w1

-",'L

'U

"'$.O L.'

"*.S 4

*p-D(S.U.)

.,7"

2.

77*'£ l#

Conductivity

,(lT

.,AIntiaalia..,

Iniia Total~AT reib al¢lori*e (mg/b)e o ontete utfl I 01adItkesmls 9___

ofmeaue(C A

95 SOP~..T0 xhbi A2.3, reiinl

-11

TVA / Sequoyah Nuclear Plant, Outfall 101 - UV-treated August 11-18, 2015 pimephalespromnelas Chronic Whole Effluent Toxicity Test EPA-821-R-02-013, Method 1000.0 Daily Chemical Analyses Projeet mnmber.

10842 wU C on cen tration P aram eterD a,D a ID 2D a 3a 4D a *D 6

pH (SUfl 7.71 7.66 7.80 7.60 7.76 7.33 7.69[

7.70 7.68 7.51 7.74 7.71 7.77 7.51 DO (m/L 7.8 7.8 8.0 7.9 7.8 6.9 7.8[.

. 7.9 7.8 8.0 8.0 7.9 7.7 7.2 Conductivity (-roes/er) 325 310 313 307 317~

313~ 1 327 i6*,

Control Alkalinity (mig/L CaCO3) 60 9

60

/

_______Temaperature

(°C) 24.8 24.6 24.9 24.4 24.8 24.61 24.91 24.7 24.9 24.3 24.9 24.6 24.9 24.6

_________SU)_____

7.70 7.68 7.84 7.61 7.80 7.321 7.691 7.44 7.59 7.50 7.77 7.71 7.8,1 7.45 TDO (nigfL) 7.8 7.8 8.0 7.9 7.8

6.

7.8*

7.9 7.9 8.1 8.0 7.9 7.8 7.2 10.7%

Conductivit mahoslcm) 310,:e***.*z.

3001 300

294

.301 306.2 316 J

_______Temaperature

CC) 24.8 24.5 25.0 24.6 24.9 24.7 25.0 24.4 24.9 24.6 25.0 24.4 24.9 24.5

__SU)7.9 763_7.85 7.60...

.0 7.37

.681 7.43 7.6 7.5 7.8 7.71 7.82 7.44 D (m/)7.8 7.9 8.0 7.9 7.8 6.9 7.91 8.0 7.9 8.1 8.0 7.9 7.8 7.2 21.4%

Conductivit (mhos/km) 293 *L!,*[

287

-~

289 28'3 304!

29l0 297 *q-*gg

_______Temperature C°C) 24.8 24.7]

25.0 24.6 25.0 24.5 25.0[

24.4 24.9 24.7 25.0 24.6 25.0 24.6 pH (SU)

"7.68 7.66 7.86 7.55 7.80 727 7.68 7.44 7.69 7.51 7.82 7.71 7.83 7.45

DO (ragL) 7.8 7.9 8.0 7.9 7.8 7.0 7.9*.

80 7.9 81I 8.0 8.0 7.8 7.2 42.8/%

Codciiy(lma/m 26~7 Yo 258 259 255 269 263

,.i 264

______Temnee--nreCC 24.8 4

5 4

25"1 24.4 24.9 24.5 25.0 24.6 25.0 24.6 nil

(~TD 7_68 7.66 7.87 7.6U 7.81 7.2*

7.7{)

Y.qU 7.70 1.3U 1.62, 1.11 1.53 I.q':t 85.6%

DO (mn/L) 7.8__

M 7.9I 8.11 7.91 "7.8

7.

8

.4 7.91 8.11

8. 8.0 1

8.0

7.

7.2 Temineratnre (0C-*

25.01 24.7 25.1 24.6 4-I I

nil f(I'T) 7.811 7.28 7.71 4-

. I DO (muIL*

~1-Conductivity (ttmhos~cnm) 100%

Alaint (malL Ca.

-I-flairdnesa (rangS, CaCO3 )

'q'otal Residual Chlorine (mW/L) remera Ir rC

.3-nl [,tgT ]I 2198 186 186 67

.*m 697 25.0 24.5 25.0 24.5 25.0 24.6[

8.0 7.9 8.1 8.0 8.1 7.9 7.2 193 190 198

/

<0.10 19 19 24.6 25.1 24.4 25.1 24.4 25.1 24.6 00 (mi'IL~

v Condgncfivitv (mumhlcms/m 100% Intake Alkalnity (nm/L CaCO 3) hardness (malL CaCO3) remnerature (°C)

I I

r l

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

File: sqnlO1._.O8111 5chem-UV.xlsx Entered by. J. Sumner Reviewed by:

t' Eeiwimun ental Testlng 5dmu'*niInr,.

Page ____

Page 1

of 2*-

Analyst ~

Date analyzed ~

Alkalinity (SM 2320 B-1997)

Matrix: Water, RL --1.0 mg CaCO2/L Titrate samples to pH = 4.5 SJ.U Time completed L -0 Z.

X]

l1trant lZnormnih and multdili~r dpltoriinidionm I

IpH of Normality Normality (A') of H2zSO4 pH Factor or Multiplier SDeionized Titrant check Begin End Total

-- (5 ml Na2CO3 x 0o05)/E

= (N x 50000)/100 ml sample Swater reference standard ml ml ml 0.25/E

= N x 500

=14.5 S.U.

number number

.. E)

(aceeptable range-- 0.0180 - 0.0220)

__.___ttq t*

o.i i,2.2, 12...

,O.02 "

,,10.2 rReference standard True value Sample

.Alkalinity (MV)

% RS=MV/TVx 1D0 S

number (TV) volume Begin End Total Multiplier (mng CaCOs/L)

(acceptable range 1(mug CaCOsiL)

(ml) ml ml ml

= 90 to110%)

Duplicate sam ple precision:

Sample Alkalinity J

% RPDj=

Sample Sample ElD volume Begin End Total Multiplier (mg CaCOa/L) t(S -D)/I(S+DYt2]}x 100I number (m1) ml ml ml ace tberange =

10%)

Duplicate (B)

"I***

l*,

j D

.01#*

Matrix sp,,ke reecovery:-

Reference standard Spike value Sample (spike alkalinity (A) number (SV?)

volume Begin End Total Multiplier (mug CBCO3IL)

(ma CaC*iL)

(ml) mi mlm.

I II I

I Sample alkalinity (B)

IMeasured spike value (MV)

% R=MV/SVx 100 I

I

.foran...,,P ranG.

I (ma CaCO/L,)

=

';Sto.12S%

I 1~

02.02.

.qamnip measunrements.,

Sample volume Begin End Total Alkalinity Sample number Sample ID (m)ml ml ml Multiplier (m CaCOIL) 1.O0

8.o qf.*9 e.O8AUM!

£4....

t¶,

t,.

f IiX*

z

.0 J__

Reviewed by Date reviewed:

O&.l*,'1I Page 61 of 95 Page61 o 95SOP C6 -Exhibit C6.1. revision 01-01-13

TE-'r """...

F I -'..

I, Em'enm~ta MTesting SolIWon shic.

Date aaye iT~rant normnality and rm Page _____

Page 2 -of 2 UltinlUer determinatim Alkalinity (SM 2320 B-11997)

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

Time completed pH of Normality Normality (Al) of 112S04 pH Factor or Multiplier

.. Tirnt cheek Begin End Total

=(5 nl Na2CO3 x 0.05)IE

= (Ni 5 0008)1100 ml Sample water reference standard mii mii!-

T.2/

N 0

--4.5 S.U.

number number (E)

(acceptable range = 0.0

-gl02*

tt,,

Laboratory c omrol standard:________________________

Reference stnad True, value SapeAlkalinity (MV)

% RS =MV/ITV x 100 number (TV) volume Begin End.

Total IMultiPlier (rag CaCOsIL)

.(acceptable range

.(rag CaCOa/L)

(ml) ml ml nml

= 90 to 110%)

Duplicate sanm l~e precision:,.____

Sample

" Alkalinity E

%RWD--

Sample Sample ID volume Begin jEnd Total Multiplier (nig CaCO3/)

((S - D) /f(s+D)*2]} x l00 number (ml) mi ml ml (acceptable ra__ge =-__10%)

Mariplie ecve(B:

llfrnesadr pk ~le Smi pk laiiy Anumber (SV) volume Begin End Total Multiplier (rag CaCOsIL) 1~

ample-algallnil*'-(lfg__ [---.lweasur.ea_*pi* wv)

[

/* * = ff*v / *v x *u*

I fln*

I I

I.h::

I-~ampIe~anca1mnazy4n1ueaaureaspaIeimma4.mY4.......I

~ it = IVIVI ~V I lUll U

I (rag CaCO3/L)

MV =A -B (mc CaCOSL)

I (acceptable range

= 75 to 125%)

I Sampl measurements:

'Sample volume Begin End Total Alkalinity

_Sample number Sample ID (ml) nl ml ml Multiplicr (ra CaCOIL) 1.I-2-

3zI

t**~z*,t*

O*

.*\\O~

3 *

e.*

.3*(

,'c

,N.

oe

... 3 50.*..

,'1,) 3.

J Reviewed by:

EZi~IIZ Date reviewed:~ i U*'i-IIII[

Page 62 of 95 SOP C6 -Exhibit C6.1. revision 01-01-13

I...

I

EnvlnmenottuITosting SolutmIonslc.

Page (oZ.

Hardness (SM 2340 C-1997)

RL =1.0 mg CaCO3/L Date analyzed J*

j Titran~t normaikv and vmultplier determination:

Page

of J Time completed [, jj0j Titrant Normality check rBegin End Total Normality (N) of EDTA pH Factor or Multiplier reference standard ml ml nil

-- 0Z(E, (N x 59009) 59 nil sample number number CE)

(acceptable range = 0.0180 - 0.0220) I N x 1000 itJ 126i o.0 io~o

o.O

.02.70

,o.oj

[Reference standard True value Sample Hardness (MV) "% ItS = MV TV x 109 number (TV) volume Begin End Total Multiplier (mng CaCOsIL)

(acceptable range (ragCaCOs/L) sm) l mi jml I

0t 1%

Dpite Ver lo" Samplel J

ardness I % R P DI-Sample Sample ID

[volume IBegin [End ITotal Multiplier (mg CaCOsIL)

[ (

54-D)ID)f2]}.xl00 number ml ml ml Matrix spike recovery,:

Reference standard Spike value' Sample Spike hardness (A) '

number (SV) volume Begin End Total jMultiplier(rgCO/)

nubr (rag CaCOsIL) ml) lm l

jml COIL I

Sample hardness (1B)

(rag CaCOsIL)

Measured spike value (MV)

MVf=A - B

% R =MV/ SV x 100 (acceptable range

= '75 to 125%l I

I.

p4 QO., 67 I

I Sapemeasurements:-

S"Sample volume Begin End Total Hardness Sample number Sample ID

____(m.

ml salj.ml Mutlier (ran CaCOs/L)

TV =ND Bsoldn bO=0 X)a0/L 0.0 o

, 04

003 OMY-.Io,*

,.(i '1.3 S

.x.,s,

]

, 3O~

4,a i

___3 8o

su*

r\\ 4

....... z.%

4.

Note: ff >l~ml of titrant is used, sample must be diluted.

Reviewed by:

LUZ Date reviewed 0 *-

Page 63 of 95 Page63 o 95SOP C7 - Exhibit C7.1. revision 01 13

"ST S

SEnvironmetl Tedting !coutions, hIn..

Page

3*

Page ____

Hardness (SM 2340 C-1997)

RL = 1.0 ing CaCQ3/L Analyst f 7.

raiat normailty and mnultiplier deters, Time initiated FC*

[T Time completed L

~

I Laboratory control standard:

Reference standard True value Sample IHardness (MV)

-% RS = MVi/ TV x 100

"number I (TV) volume Begin IEnd ITotal Multiplier (rmg CaCOs/L)

(acceptable range (ragCaCO.,/L) I Jl) mjl ml ml 90 to 110%)

I**.t36',.,40 5o

0 I~ *o Zo.o

.10.,...

06I)-

Duplicate sam lie precision:

Sample Hardness

%RPD =

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

((S - D) /j(S+-D)/2} x 100 number

f (ml)

ml ml" ni

____l_____

Matrix spike recovery:

Reference standard Spike value ISample Spike hardness (A) number (SV) volume Begin End Total Multiplier (mag CnCOaIL),

I(ragCnCO*iL) j*m!l*

ml m ml I

Sample'ihard'ness (B)

Measured spike value (MfV)

% R = MV/ISV x 100 (rag CaCOz/L)

MV -- A - B (acceptable range

__.,_____(____g CaCOaIL)

--75 to 125%)

Sample measurements:

Sample volume Begin End Total Hardness Sample number Sample Il)

(ml) ml ml ml Multiplier (rgCC*L I-ND Blan.'

.5 U U mL a i*.x*

_*.J,*o,,

A2.*

l.6 3

L 212. J4_,,,

Note: If' >15ml of titrant is used, sample must be diluted.

Reviewed by:.

Date reviewed [O*.*

Page 64 of 95 SOP C7-Exhibit C7.I. revision 01-01-13

Page.*t Page i____

of I Envirnmenltal TbitIng SDlution*,Int I

I Total Residual Chlorine (O1UON-1977)

Matrix: Water, RL =OJ10mg/t Meter: Accumet Model AR25 p11/Jon Meter Analyst Date analyzed 1o{

/

leide reagent:

/t*

I I

0.10 rnglL 1.00 mng/L

'sflOfye~ted.rt~inc'- -=r2&tn~

m 1

Note: For samples with a residual c-hlorine of > 1.0 rag/L, the samples must be diluted to be within the calibration range.....="""

Laboratorj' control standard:

Reference standard

.True value'(TV)"

Measured value (MIV)

% RS -MV/

T

,i100.*

number (rog/L)

(mg/L)

(aceptaiLble rr..a~uge.= 90 to.1101%),

Duplicates smle preision:

Sample I.Sample ID

[Sample characteristics

[Residual chlorine RPD- = (S D) i)[(S+D)12fl x beO number

L-Sample measurements:

Sample Sample ID "Sample characteristics ResidUal chl0rine number nmL)

~~~~Reagent Blank

0 O

1 11 I

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

Reviewed by E tZZIZ..I Date reviewed. [I 0

.!5IJ*

SOP C8 -Exhibit C8.I, revision 01-01-13

.. Page 65 of 95

S...: Page. q

ag.L of.

.:/

':'" :. :" "-;[; i;. "

i." Total Residual Chlorine (ORION-l[977)

":i-i.

Matrix: Water, Rb -0.10 mglL

- .,.*:,.,,*i.

Meter: Accumet Model.AR25 pH/Ion Meter

.1h 0dide:

. +,,,.

i

"" i;.,,,,,*.

. r,

S..0.10 mgI]L 1.00 mg1L I..

ij :IVS I..3 147IV.~3 L~A~

I.V 2*.*

.I.1,hO. :::4...*03 i *

v

.. v

o*

,i

.,,.**i.:,,.o.*.'

i-

',*-I¸.

FSg.~

.0 gL the mailsrust be dfiuted io be within the calibra 4,::*......

bw~lieate samp~le precision:

.4[

Duplicate Samle measarements:..

V,,

Ua 1..

Sam~ple nunmber

-Sample ID I

Sample characteristics eaj ent Blank LTP~

te, (kci Dar4

":4::'

.a ---

~

4 414i%

i'tLLu,L~4 J

~

I ~

(~u~e

cdo, (je&e

.lar.rk~icft.:

\\ *.

,d.

r Ldeo*

I~~~.iPL~~O

~

0(lPCe~

," "..' :i I

I I

Ctur I

-I m

ll-Note: All samples were analyzed in excess of EPA recommended holding time (15 minutes) unless other*rwise noted..:.-'".....

o

' :.: :., *:i:

(ac,..,

e:;1,il* le aage; 9 t1....:%

Review" ed.

1: .

by

ll*.::.*,~;7"7 Date:..::

re".vi'e ed....

S

--rOP..

C8 Exh.ibit-- C,-.8.I rev.-**;*ision 01.0.1 Page,6 o 95

rage _.J.

Page __

o i~~- ~Eb nnuntal Testing Solutions, Inc.

k Analyst 7~7 Date analyzed ~

[*otal Residual Chlorine (ORION-1977)

Matrix: Water, RI = 0.10 mg/L Meter: Accumet Model AR25 pH/Ion Meter 0.10 mn/L

[

1.00 mEIL (snetd an 00mV* I

,tandaid. numiber (mY)

I

" I

'1 II I i'r 1

i I

I

" lift--

llml ii iI rsamples with a residual chlorine of > 1.0 ing/l, the sples mustb oebe within the calibration range.

i-

.*L aboratoTp eontrol standard:......

(Jb (5 I Ii ',"

I In flnnlieatR s~amnIA nrac!ISIoA:

Sample Sample ID Sample characteristics Residual chlorine

%RPD = {(S - D)/I[(S+D)12]} x 100

] number D~upliat D*

4)*"

.j *l Samlemesurements:..

Sample

-Sample ID Sample characteristics Residual chlorinei number t*L.

~~~~~Reagent Blank

.*t" Dd. VeiIe~, L. -

z ze q3 McG-hIge 001 I~O Color d~qv

~ati~

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

Laboratory~control standard:

Reference standard True value (TV)

.Me asured value (MV)

% RS=fiMV /TVx I00 fvs

~~05

.number (an./)

(mg/L)

(acceptable range = 90 to 110%)I Reviewed by t'U(

Date reviewed l

!t SOP C8 - Exhibit C8.1, revision 01-01-13 Page 67 of 95

Page of "BnuvronwentaI Testintg Solutlon;, mt.

Total Residual Chflorine (OmION-1977)

Mat'ix: Water, RL = 0.10 mg/L Meter: Accumet Model AR25 pH!Ion Meter

'i*!:

Analyst 7

7i

Date analyzed fI--0"**t 3

14odide reagent: IAI]*

Acid reagent:

,hRe r,

i 0.10 mg/L 1.00 mg/L mV Change (suggested range = 26 to 30 mV) I

..Millivolts (mV),

,'..,+.

'l[

l lll ll*

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

Laboratory control stun dard:

Reference standard True value (TV)

IMeasured value (MV)

% IRS MV/T"V x 100 number (ra~g/L)

I(rag/L)

(acceptable raage=ff 90 to 110%)1 D lctesm le recision :

[-am-e Smple ID

.Sample chrateisi s

aml eaaeite Residual chornhPDori-n-eI(+D12)x 0

number accptbl r manIe,

1 Samagent Blans"re, ents:

Note: All samples were analyzed in excess of EPA recommended holding time (1 5 minutes) unless otherw~ise noted.

Laboratory control standard:.....

[

Referenceestandard True value (TV)

Measured value (MV)

% RS--MV /TV x100 1 number (mgIL) -

(mg/L)

(acceptable range = 90 to 110%)

Reviewed by LZ tIII IIII Date reviewed [..O...d$-.j......

SOP C8 - Exhibit 08.1, revision 01-0.1-13 Page 68 of 95

Sequoyah Nuclear Plant Biomonitoring August 11 - 18, 2015 Appendix D Reference Toxicant Test and Control Chart Page 69 of 95

() Environmental Testing Solutions, Inc.

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

1.0 0.9 0.8 0.7 I

I I

J j

I I

I USEPA Control Limits (+/- 2 $tandard Deviations)

I I

III I

I 1

I I

0.56I 0.5 1,4 S1.2 m>.0.8 S 0.6 I',

UEAWarning and Control Limits (5ad90 hPercentl CVs)

I I

I II I

I I

t I

1 I

I II l&

,,, A

.'rI I

I i

I I

Ii I

I I

f Ii J

i 1.2 -

Laboratory Warning and Control Limits (10t and 25h Percentile O~s) 1.0 GA t

r*

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

Central Tendency (mean IC25)

Warning Limits (mean IC25 + SA10 or SA.75)

Sraphsgensrmtedf...

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

Rxevl:ewdhte~edbw;.:me spruadsheet.

Page 70 of 95

-J Envlronmental Testing Solutions, Inc.

0 Pimephales promelas Chronic Reference Toxicant Control Chart CO Test number Test date 7-day ICs CT

.(g/L KdI (g/2KCI)

State and USE S

Control Umll cT-25 CT.

1.

2.

1 1

1 1(

06-08-14 0.76 0.76 0.05 0.659 0

2 06-10-14 0.79 0.76 0.05 0.65 0.1 3

06-19-14 0.76 0.76 0.05 0.65 0.1 07-08-14 0.74 0.76 0.05 0.67 0.5 08-05-14 0.86 0.75 0.05 0.66 01i 6

08-12-:14 0.82 0.77 0.05 0.67 01*

7 09-09-14 0.81 0.77 0.05 0.57 0.1 g

10-07-14 0.77 0.78 0.05 0.69 01i 9

11-04-14 0.82 0.78 0.05 0.69 0.1 0

"12-02-14 0.74 0.78 0.04 0.69 0.1

2.

01-13-15 0.76 0.78 0.04 0.70 ThI

2.

01-21-25 0.81 0.78 0.04 0.70 0.2 LS 02-OS-IS 0.74 0.78 0.04.

0.69 0.1 L4 02-03-25 0.78 0.78 0.04 0.69 0.2 L5 04-07-15 0.81 0.78 0.04 0.71 0.2 L6 05-05-15 0.82.

0.78 0.04 0.71

. 0.1 17 06-09-15 0.82 0.78 0.04 0.71 0.2 18 07-14-15 0.78 0.76 0.04 0.71 0.2 19 09-04-15 0.88 0.79 0.04 0.71 0.2

.0 08-11-195 0.82.

0.75 0.03 0.72 0.1 Note:

7-d lC* = 7-day 25% inhibition contentration. An estimation oftho cs CT= Central tendency (mean IC~sl.

S =Standard deviation of the lC~n values.

Laboratory Control and Warning limits Laboratory control and warning limits were establshed usir recommended isy USEPA for the test method and endpoint.

5a.so = Standard deviation corresponding to the 10"' percentile CV.

5A.25

- Standard deviation correspondring to the 256 percentile CV.

USEPA Control and Warning Limits S.s= Standard deviation corresponding to the 75*~percentlle CV,

$.so= Standard deviation corresponding to the 90* percentile CV.

CV = Coefficiernt of variation of the ICan Values.

A O5 2S 0.09

~6 0.09 0.03 0.09

~6 0.09

~7 0.03.

7 0.09

~7 0.09

~7 0.09 i7 0.09 0.09 0.09

)6 0.09 0.09

)6 0.09 6

.0.09 6

0.09 6

0.09

6 0.09 Laboratory Warning Umits CT -S*

Cr + S* 0 Laboratory SA.ns Control Umits CT-S*.

C'+ S*

USE PA SA.TS Warning Limits CT - 5S.n5 CT +/- SA.7s USEPA S*s Control Limits CV CT-S*.

CT+ Spre 0.66 0.67 0.66 0.67 0.67 0.68 0.68 0.68 0.68 0.69 0.69 0.69 0.69 0.69 0.89 0.69 0.69 0.69 0.69 0.70 0.85 0.85 0.85 0.85 0.86 0.86 0.87 0.67 0.87 0.87 0.67 0.88 0.87 0.67 0.88 0.85 0.88*

0.98 0.88 0.89 0.18 0.60 0.16 0.60 0.16 0.60 0.18 0.60 0.16 0.60 0.16 0.61 0.18 0.61 0.2.6 0.61 0.2.8 0.61 0.2.6 0.61 0.16 0.62 0.16 0.62 0.16 0.62 0.16 0.61 0.2.6 0.62 0.16 0.62 0.16 0.62 0.16 0.62 0.16 0.62 0.17 0.62 0.91 0.29 0.92.

0.29 0.91 0.29 0,92 0.29 0,93 0.29 0.93 0.29 0.94 0.29 0].64 0.30 0.84 0.30 0.94 0.30 0.94 0.30 0.95 0.30 0,94 0.30 0.94 0.80 0.95 0.30 0.95 0.30 0.95 0.30 0.95 0.34 0.95 0.30 0.96 0.30 0.47 0.47 0.47 0.47 0.47 OA8 0.48 0.46 0.48 0.49 0.48 0.49 0.48 0.48 0.49 0.48 0.49 0.49 0.49 0.49 1.04 1.04 1.04 1.05 1.06 1.08 1.07 1.07 1.07 1.07 1,08 LOS 1.06 1.07 LOS8 1.08 1].08 1.06 1.08 1.09 0.34 0.42 0.34 0.42 0.34 0.42 0.34 0.42 0.34 0.42 0.35 0.42 0.35.

0.43 0,3.5 0.43 0.35 0.43 0.35 0.43 0..96 0.48 0.35 0.43 0.35 0.43 0.35; 0.43 0.35 0.4.8 0.35 0.43 0.35 0.43 0.35 0.43 0.35 0.43 0.26 0.43 1.10 0.07 1.2.0 0.07

.1.10 0.07 1.10o 0.09 1.11 0.07 1.12 0.07 1.12 0.07 1.13 0.06 1.13 0.05 1.13 0.08 1..13 0.05 1.13 0.05 1.13 0.05 1.13 0.05 1.14 0.05 1.2.3 0.05 1.2-4 0.05 1.1.4 0.05 1.2.4 0.05 1.1.5 0.04 ncentration of potassium chloride that wouid cause a 25% redaction in Pir-nephales growth for the test popsuiation.

the stsedard deviation oftheilC,~ vlues corresponding to the 10th and 25th percentlileCVs. These ranges are more stringent than the control and warning limits (SAIS= 0.12)

(SAsn = 0.21)

(SAsS = 0.35)

(S~so= 0.45)

UStPA. 2509ULnderntandlngand Arosantingfar Method Variability inWhole EffentTosxcltAppliratilnos Ilderthe National Polautant Disoharge Elimina~ien Program. tPA-933-R-50-e3, US tnvironmental Protection Agency, ndinnati, OH.

File: pphc tcr_0ll 8

f15sx Entered byc.J_.umner Reviewed b

SEnvironmental Testing Solutions, Inc.

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

1.25 e;~1.00 I,-

0.750 o0.25 I I I

I I

I I.

1 1

1 I

I I

I ".1 USEPA Acceptance Criteria (> 0.25 mg per surviving larvae)

I I

I i

i I

I I

i I

i I -,

04.. =

C O 0

30 20 I

i I

I I

i 1 -

- I -

I I

I -

I Kentucky Acceptance Limit (< 30.0%)

-. I I

I I

1 I

I I

I I" I I

I I

10 0

I I I

r I

I I

I

I I

I I

30 USPAUpper PMSD Bound (90&h percentile <30.0%)

20 10.'

I.

I I,

I,

I I

I I,

I I

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)

.u,.,

Control Limits (mean Control Growth, CV, or PMSD _+ 2 Standard Deviations)

"*%*Page72 of 95 Grmphs genoerted from eseocTated.ecel spreadsheet.

Excel spreidsheet entered by: J. Sumner Rev~ewld bm:

0

, Enel,......

tin S, t

uon,, I,,°....

Chronic Control Control Mean Test number Test date Suval Got

)f Endpoint Measurements mrephales promelas Reference Toxicant Data clr cv CT MSD PMVSD CT for Control Growth

(%)

(mg/larvae)

(m/ave

(%)

for Control Growth CV (%)

(%)

forPMSD (%)

1 2

3 4

5 6

7 8

9 10 11 12 13 14 15 16 17 18 19 06-03-14 06-10-14 06-19-14 07-08-14 08-05-14 08-12-14 09-09-14 10-07-14 11-04-14 12-02-14 01-13-15 01-21-15 02-03-15 02-03-15 04-07-15 05-05-15 06-09-15 07-14-15 08-04-15 100 100 100 100 100 100 100 100 100]

100 100 100 100 100 100 97.5 100 100 100 inn 0.672 0.723 0.715 0.631 0.560 0.694 0.622 0.617 0.627 0.712 0.551 0.540 0.585 0.6.07 0.520 0.536 0.534 0.556 0.539 I, *;Aflt 0.697 0.703 0.685 0,660 0,666 0.660 0,654 0.651 0.657 0.648 0.639 0.635 0.633 0.625 0.620 0.615 0.611 0.607 6.2 10.3 5.9 8.3 9.0 2.6 11.7 11.6 5.6 8.2 8.7 10.9 13.7 5.5 4.3 13.0 2.5 4.6 11.9 8.2 7.5 7.7 7.9 7.0 7.7 8.2 7.9 7.9

'8.0 8.2 8.7 8.4 8.2 8.5 8.1 7.9 8.1 0.10 0.11 0.09 0.06 0.07 0.11 0.10 0.11 0.05 0.09 0.06 0.08 0.08 0.07 0.06 0.07 0.05 0.10 0.07 14.2 15.9 13.2 9.0 13.3 15.6 15.7 17.2 8.6 12.6 10.8 14.7 13.5 12.2 12.1 12.7 10.2 17.5 12.1 15.0 14.4 13.1 13.1 13.5 13.8 14.2 13.6 13.5 13.3 13.4 13.4 13.3 13.2 13.2 13.0 13.3 13.2 Note:

CV = Coefficient of variation for control growth.

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

Upper CV bound determined by USEPA (90th 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 end treatmentthat can be declared statistically significant In a whole effluent toxicicy test.

Lower PMSD bound determined by USEPA (l0'm percentile) =12%.

Upper PMSD bound determined by USEPA (90C percentile) 30%.

CT -- Central Tendancy (mean Control Growth, CV, or PMSO)

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. 200Ta, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-termn Chronic and Acute Whole EffluentToxicity Test Methods, Volumes l and 2 Appendix. EPA-g21-B-01-O04 and EPA-g21-B-01-005.. US Environmental Protection Agency, Cincinnati, OH.

Page 73 of 95 File: ppkclcr_081 115.xlmt Entered by:.1. Sutmner Reviewed by:

°0T 0@

Page 1 of 6 Potassium Chloride Chronic Reference Toxicant Test (EPA-821-R-02-013 Method 1000.0)

Species: Pimephales promelas PpKCICR Test Number: 3041 Dilation preparation infor~ma~tion:'..

Comments:

KCI Stock INSS number:.

TINSS s"AQ________________

Stock preparation:

50 g KCII:......

_____________________Dissolve 50$KIi MliQwtr Dilution prep (r. g/L) 300 450 600 75019001 1050 Steek volume.(mL) 6 9

12 77577 18 21V~F ____________________

Diluent volume (mL) 94 991 988 985 982________________97______

Totalivolume (mL) 1000'100" 1000 1000 1iJ2000

.1000 p

Test org~anism in formation:

ITest information:

-~

Date and times organisms were born between:

~O~I Incubator number and' shelf location:

Organism source:

ATOX Batch Pp: O!*t *.i*

Artemia CHM number:

j CHM836 Drying information for weight

_____________________ _______________________determination:

Transfer vessel information:

pH = "* d'a S.UJ.

Date / Tinme in oven:

I o-llt¢i' io~4q

____________Temperature.

.R

"*a*

°C.

initial oven temperature: I

!hIC~.

Average transfer volume:

0.1749 nmL Date / Time out of oven:

el *1 bt Final oven temperature:

I' Daily feeding and renewal information:..

Day Date Morning feeding Afternoon feeding Test initiation, renewail, or termination MHSW batch used Time AnalyVst Time Analyst Time Analyst 2

08-11-15 4b4{

JS'*,

o 4-(

t*l"I 1

08-12-15

.4.t*

A"t"b*

I O3

Ct.O-K 08-14-15 (Y0*

H l'O I lOO&

g

.CaIgI" S 1) 6 08-17-15 AU' LDOU*

7 08-18-15 o "

v V.

Control information:

Acceptace criteria Sunmary of test endpoints:.

% Mortality:

01[.

_<20%

7-day LCso

'?. to SAverageweight perinitial larvae:

oiO.I0 NOEC t

Average weight per surviving larvae:

O-¶aCO0

>_0.25 rag/lIrvae LOEC ChV

!,;'tO. R CIIV(Ji1 74 of 95 74 of 95SOP AT2I - Exhibit AT21.l, revision 11-01-14

Fflnvlmn mwrtjahliTgdmfn

uZo n* Ir Page 2 of 6 Spiecies: Pimephales promelas PpKCICR Test Number: 30._4 Survival and Growth Data Day Control r_

300 m* KCIIL 450 mg KCI/L A

B" C

[D E

F G

H I

J K

L 2

i0 10 li0 o

I

/O tO t IQO

!001 3

(0 1o 1o~ ~6jto

/0 1O J10 I0 l0

/O

/0 R~l~.tZ II~

I 4

I0 to to I0 t '

/0 10

'O (0

'0

/0 go ty I l (0t10 JO tO (o 0 1l*

1O0 /0 ~

6 I

t l10 10 to0 1D 0

L.o 10

/0 l0 7

tb

/(t j1 tI 10 I0 t

l*

O A=pan weight (mg)a eu.

Date:

B-=Pan +Larveweih (ra) 33 Dat:

t e:*--

-,-t--

C = Irvae weight (mg) =B-A I-I I

I~~~1fl~f.

1 -

~

U Hand calculated.

Analysft:

1.11..

S. ~%

i?

t..;

f :

I

Weight per initial number of larvae (mig)

= C /Initial number of larvae Hand calculated.

Analyst.:

N ci.)

Q.

0 N

ft,

'V N

0 c#

0*

0" C,

Average weight per initial Percent number of larvae (rag) reduction

_______________________rom coto SoA*'6'5

-*A,7 1 0.*'

.7 Comment codes: c = clear, d =dead, fg =fungus, k = killed, m = missing, sk = sick, sin = unusually small, Ig =unusually lar'ge,.d&r = decanted arnd returned, w = wounded.

~ omment.

SOP AT2l - Exhibit AT21,1, revision 11-01-14

0T Gn@ n,~Iet~o~~,Ic Page 3 of 6 Species: Pimephales Dromelas PpKCICR Test Number: 30_4 Xurvival and Growth Data Day 600 m* KCIIL.

750 mg KCI/L

__900 mg KCI/L M

N 0

P; Q

R S

T U

V W

X 010 10 10 10 10 10 '

10 10 10 10 10 10 1

IiQ 1Ot10 1010

/__ t_

___/

2 I) i',(

tO ibIOl tI1IO i 10 1g 0

3 1t0 10 10 l0 10 10 4

I-4~-----4 I--------t~------~l 4

10 I0 10 7

....,1*

W i

A =Panweight (,ag), *s Tray color code::

111

-ne,.~iA I~

4i~I'~

I~~llr~

~

Date: 0* a Dat:..

C = Larvae 'weight (mc* = B-A I ~

js~~f 4~l ~I5.1I~.~I3A' h.ai¸~'

Hand calculated.

Analyst:.

Weight per initial number of larvae (nig)

SC / Initial number of larvae I

a a

I

[

n

.a Average weight per initial Percent number of larvae (iag) reduction 0

q..*

1],

O S' o0.V?'.

O" °*1 t'-Io.*[]

from

___ontrol_

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

Comments:

ie 76 of 95 SOP AT2I - Exlbbit AT2l1., revision 1 1-01-14

i"'*

1 i

i.

i i

En~vlr en! t~TedIllrehg tow.eQ Inc..

Page 4 of 6 Species: Pimenhales n~romelas PpKCICR Test Number: 30__4 i.J A = Pan weight (rag~1 Tray color co~c::

I[,hq-H*

Analyst:

lvi Date: OR-tfl-I*

B =Pan +Lar~ateweighit (rg)

Analyst:

J Date:

{*

C=fiLarvae weight (ng) -B-A AaytHand.

calculated.

A Weight per initial number of larvae (rag)

- C /Initial numnler of larvae A

lytHand calculated.

Average weight per initial Percent number of larvae (mig) rcductioun Ifrom control

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

SOP AT2I - Exhibit AT21.1, revision 1 1-01-14

-Environmental Testing Solutions, Inc.

Pimephales promelas Chronic Reference Toxicant Test EPA-821-R-02-013, Method 1000.0 Quality Control rerificatlon of Data Entry, Calculations, and Statistical Analyses Test number:

304 Test dates; Anust* 10-18, 2015 comtnbllroe(mg/LKXI] R~pOSct. enrtol numhef Steelremberef A= po~we.Fz1(mg)

B:pen~aPa rue.

SceIght (el =

Wall~tI~oeone M~un V/suredini ceo)elT[ofodrldlon Wleihtl/InP.Jl numbref 610,undref l{*i Menuen~ahtJ'Iel.

6.fn1dl.,temr/lltlen Pemntremu~f ste ieee

" loe.

nmrl~t mel 8.8 ncabreftereflrseeleg enrmbereFlerr(mg) I6ll~'m~e eamgl nurnberrol(ureo

16) eneeol {p4 A

10 10 16.05

. 21. 2 5.07 0_357 0.537 B

10 10 16.19 22.*

8.34 0.634 0,600 7.8 0.634 100.0 0.603 7.8 Not applicable c

13 10.

18.28

22. 60 5.90 0-530 0_330.

10 10 313.43

19. 0 8.37 0.637 0.837 6

153 10 16.41 23.*

5.72 0.672 0.672 30 10 10 16.35

23. 86.31 0.631 o,63

5.

0.631 100.0 0.633 5.4

-3.6 (0

10 10

18.12 22.d1 5.03 0.339 0.366 14 10 10 2.5.66 722 6.40 0.640 0.630 I

10 10 18.19 22.,*

6.14 0.614 0.614 430 1

10 Ir 16.55 2.1*65 0.583057 5,6 0.353

100.0 0.667 5.6 5.4 K

10

10 16.33 21.9 5.59 0-569 0-559 L

10 10 15.01 20,,,

5.43 0.543 0.343 M

00 10 16.70 22.37

.67 0.567 0.587 6010 N

10 9

1.6.07 21.46 5.39 0-593 0.805 6,0 0.539 93.0 0.574 4.7 4.3 o

10

10 25.67 21.43 6.01.

0.601 3.801 P

10 9

34.24 20.$3, 5.839 0.854 0.589_______

-Q 10 8

55.43 20-*

4.87 0,603 0.487 Pi 108 1.7 20J*

5.3080.630 3,

.63 80.0 0.535 7.1)

+/-0.7 5

10 9

16.20 2L2.

5.72 0,636 0.572 T

10 9

14.05 20,2 5.,7 0,619 0.557 U

10 5

13.84 19 1 3.47 0.694 0.347' 903 V

10 S

15.06

16. 4 3.16 0.636 0.623 10.2 0.318 67.6 0.357 8.7 40.6 W

10 7

16.23

20. *1 3.78" 0.540 0.378 "X.

10 8

15.77 19.1 3.86 0.643 0286 Y

10

- 2 15.59 16,9

,0.90 0.850 0.030 1050

.=

10 1

035.20 16./J 0.95 0.350 0.569

+/-9.6 0.035 12.5, 0.0,5 17.,

86.5%

PA 10

2.

16.85 17.6) 0.73 -

0.730 0.075 8

10 1

15.26 15.*

0.85 0.660 0,066 Ounnett'.s MOO value:

PM312:

0.0530 9.7 MSD=

pMSD =

Minimum b8i~ntcant Difference Percent Minimnem Significa nt Difference PMSD (so /I1eusure of test precision. The PMSD is the minimum percent differenrce between the control and treatmient that nan be declared atatisticc~ly significant in a w oeeffluent tonicity 5630.

Lower PtslD bound determined by USOPA (10th percentile) =1:.

Lower andlupper pMSD bounds were detetrnined from tire 10th and 30th pertentile. respectively. of PIMSO data from EPA's WtTlnterlahoratery Variabiliny Study (USEPA. 2061a; USEPA, 200lb).

USEPA. 2000a, 2001b. Final Report; Interlahoratory Variability Study of EPA shorE-term Chronic a rd Acute Whole Eiffuent Toxicity Test Methods, Volumes :1 nd 2-Appendit. EPA-821-8-01-004 and EpA-821-B-O:1-005. US Eneironmental Protection Agenlcy, Chmcinncl, OH.

File: ppkclcr_03111l5.:,dsn Snteredi,1n.

2. Sumner Reeiewed hy,:*

0ET 0

Larval Fisen Urowtn and Survival Teat-7 uay survival Start Date:

811112015 Test I D:

PpKCICR Sample ID:.

REF-Ref Toxicant End Date:

811812015 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

KCL-Potasslum chloride Sample Date:

Protocol: FWCHR-EPA-821-R-02-013 Teat Species:

PP-Pimephalea promelas Commenta:

Conc-mgJlL 1

2 3

4 D-Control 1.0000 1.0000 1.0000 1.0000 300 1.0000 1.0000 1.0000 1.0000 450 1.0000 1,0000 1.0000 1.0000 600 1.0000 0.9000 1.0000 0.9000 750 0.8000 0.8000 0.9000 0.8000 900 0.5000 0.5000 0.7000 0.6000 1050 0.2000 0.1000 0.1000 0.1000 Transform: Arcsln Square Root Rarnk 1-Tailed Number Total Cona-mgllL Mean N-Mean Mean Mini Max CV%

N Sum Critical Reap Number D-Control 1.0000 1.0000 1,4120 1.4120 1.4120 0.000 4

0 40 300 1.0000 1.0000 1.4120 1.4120

1.4120 0.000 4

18.00 10.00 0

40 450 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4

18.00 10.00 0

40 600 0.9500 0.8500 1.3305 1.2490 1.4120 7.072 4

14.00 10.00 2

40

'750. 0.8500 0.8500 1.1781 1.1071 1.2490 6.954

4 10.00.

10.00 6

40

=900 0.5750 0.5750 0.8820 0.7854 0.9912 11.405

'4 10.00 10.00 17 40

1050 0.1250 0.1250 0.3572 0.3218 0.4638 19.851 4

10.00 10.00 35 40 Auxlllary Tests Statistic Cditical Skew Kurt Shapiro-Wilk's Teat indicates normal distribution (p > 0.01) 0.9 0.896 0.47889 -0.2624 Equality of variance cannot be confirmed Hypothesis Teat (1-tall, 0.05)

NOEC LOEC ChV TU Steel's Many-One Rank Teat 600 750 670.82 Treatments vs D-Control Maximum Likelihood-Problt Parameter Value SE=

95% Fiduclal LImlts Control Chi-Sq. Critical P~value Mu S~gma Iter Slope 11.9904 1.7058 8.64743 15.3334 0"

5.01278 9.48773 0.28599 2.95064 0.0834 3

Interceot

-30.379 5.01458

-40.208

-20.551.

TSCR Point Prob its mgIL 95% Fiducial Limits EC01 2.674

,570.98 481.208 631.152 ECOS 3.368 650.8165 74.964 701.582 ECI0 3,718 597,842 531,366 743.265 EC15 3.964 731,476 671.845 773.549 E020 4,158 759.359 705,217 799.222 E025 4,326 784.127 734.488 822.685 EC40 4.747 850.177 808.951" 890.149 ECS0 5.000 892.583 652.812 938.541 EC60 5.253 937.061 894.778 993.824 EC75 6.674 1015.99 962.914 1100.54 6080 5.842 1049.13 989.896 1147.7 685

.036 1089.12 1021.63 1206.03 EC9O 6.282 1141.62 1062.27 1284.54 EC98

.8.645 1224.11 1124.44 1411.74 E;C90 7.326 1395.26 1248.96 1688.09 1.0 0.9-aO.6*

I*0.4-0.3-;

0.2-0.0 10 100 1000 10000 Dose mg/L.

Dose-Response Plot

,1 0.9 0.8 0.7

'*0.8 (I 0,5 0.3 0.2 0.1 I..

Page 79 of 95I File: ppko~cr_05111l5.xlsx Enteral by: J. S~iner Reviewed by:

"",,nd%'mnrmertlflhsfng Saivflann%

In, Statistical Analyses Larval Fish Growth and Survival Teat-? Day Growth Start Date:

8/1112015 Test 1D: PpKCICR Sample ID:

REF-Ref Toxicant End Data:

8118/2015 Lab ID:

ETS-Envir. Testing Sol.

Sample Type:

KOL-Potasalum chloride Sample Data:

Protocol: FVVCHR-EPA-921-R-02-013 Test Species:

PP-'Pimephales promelas Com ments:

Conc-mgIL 1

2 3

4 D-Control 0.5370 0.6340 0.5900 0.6370 300 0,6720 0.6310" 0.5890 0.6400 450 0.6140 0.5530 0.5590 0.5430 600 0.5670 0.5390 0.6010 0.5890 750 0.4870 0.5250 0.5720 0.5570 900 0.3470 0.3180 0.3780" 0.3860 1050 0.0900 0.0950 0.0730 0.0650 Transform: Untransformed 1-Tailed Isotonic Conc-mgIL Mean N-Mean Mean Min Max CV%

N t-Stat Critical MSD Mean N-Mean D-Control 0.5995 1.0000 0.5995 0.5370 0.6370.

7.820 4

0.5163 1.0000 300.

0.6330 1.0559 0.8330 0.5890 0.6720 8.404 4

-1.324 2.290 0.0580 0.6163 1.0000 450 0.5675 0.9462 0.5673 0.5430 0.6140 5.616 4

1.274 2.290 0.0580 0.5705 0.9260 800 0.5740 0.9575 0.5740 0.5390 0.6010 4.748 4

1,007 2.290 0.0580 0.5705 0.9260 760 0.5353 0.8928 0.5353 0.4870 0.5720 7.038 "4

0.6353 0.8686 900 0.3573 0.5959 0.3573 0.3180 0.3860 8.707 4

0,3573 0.5797 1050 0.0810 0.1351 0.0810 0.06800 0.0950 16.958 4

0.0810 0.1314 Auxiliary Teats Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.96241 0.944

-0.261

-0.6765 Dartlettas Test indicates equal variances (p = 0.83) 0.86885 11.3449 Hypothesi$ Test (l-tall, 0.05)

NOEC LOEC ChV Tru MSDu MSDp MSB MSE F-Prob df Dunnett's Test 600

>600 0.05796 0.09668 0.00355 0.00128 0.05716 3, 12 Treatments vs D-Control Linear Interpolation (200 Resamples)

Point mql/L SD 95% CL(Exp)

Skew IC05 I101 IC15 IC20 IC25 IC40 1C00 401.30 89.61 339.92 615.13 1.6308 8657.64 98.87 286.82 820.20

-0.9283 759.64 27.79 625.04

.801.91

-1.3380 785.60 14.63 735.32 823.90

-0.5965 611.57 12.60 770.97 847.30

-0.2674 889.47 10.93 866.23 920.03

-0.1488 926.67 8.76 904.09 946.21

-0.3354 1.0T 0.9 0.8 0.7 G.6 I

a 0.4' 0.3' 0,2' 0.1' 0.0, t

~1-

-0.2 0

,800 1000 1500 Dose mgIL Dose-Response Plot 0.8 0.7 0.6

0.5 0.4 093 0.2 0.1 0

Page 80 of 95 1-tall, 0.05 level of significance o

a a

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nlnena~~fgS~~o*lc Page 5 of 6 Species: Pimephales promelas PpKCICR Test Number: 30 Daily Chemistry:

Day

v. tnerformed toH. D.0. and conductivity measurements only.)

(Analyst identified for each d,"

o_

1 2

Analyst i

- I A/&

ILA/6

/

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Concentration Parameter nIlv (S.U3

-~-~

IILI~

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'-/oq&

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~

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II A

Conductivity

&unhnslcm)

Mkahlnjty (mg CaCO3IL)

!~

CONTROL 367 II7.4 1

"Herflflooc V

(inn CaCO3IL)

Temperature (0C)

",k I

Li.Io 4

~

I.

pH (SU.)

7. _'/.

'/.92*

7 7~

300 mag KCI/L DO" "m

L I

f

.11 "

I*L..

....~i~

Conductivity____

011 Qiahscm

Temperature (0C)

'A.S.

13~A 1341~

I#zs' p

41

3.

pH (s.u.)

DO (maglL) 450 mg KCIIL Conductivity llIt:a 17, j

Temperature (°C)

"U4. A -ii ~iA.S I 2~4A l

pH (S.U.)

i-"i.z i.x l7~

7,8

.'b mV 4-~---~--il Do (mgfL) 600 mg KCI/L Conductivity I,-* -

7,,.

-I-

t-mtnnqln cr/m-------

-It

Temperature (°C) i-

[.*

D.(O~n) 72 7.8 7,

7.$o q*

750 mg KC!JL Condtviy

.Temnperature (C°C

.1

5A-*

",.q-.

"/*0Ao.,

pHn(s.u.)

q 7.~

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Tempeatr/L

()

"7W-7.

.0

7.

"~

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,_ Temperature (°C) 2*

"A'

,7.

"IA. to

]l,.

.1

'V L.

Initial Final Initial Final Initial Final hdoknity and hardness performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by:"_...*prtrspromda h ieo etiiitornwlo emnto yteaaytietfe nteDiyRnwlIfraintbelctd*

ae1 Page 81 of 95 SOP AT21 - Exhibit AT21, 1, revision 11-01-14

t Ervlmn m! *Teitngg o~tu

Ita*~j Page 6 of 6 Species:,Pimephales Dromnelas PpKCICR Test Number:. 304 Y

Day (Analyst identified for each day. n~erformed nil. D.O. and conductivity measurements onlv.A 3

4

5.

6 Analyst 11114 I IV.

IIlA AI/t /LW

/L 6 I1 MA Conenraio IParameter IDH (s.u.)

i~Sjg~~7,Lfl I 7,*Z' II 7.*

I 5 L I

I!."

DO (mtogL.)

CONTROL Conductivity

(~crnhosIcm)

Ajl~ahlijjty (mg CaCO3/L)

Aliartiness (mg CaCO3IL) a.'0*

II TA n

7..,,"

5Teiperatire "t4.l 300 mg KCI/L Conductivity 450mg KCil*

Condnctivity I*Oq il*

.l

)

o

)

Temperature

~4 A

~

a.

-t*'

.t' DH (S.U.)

"J.PW "7.JJU 1.'7L1*

1,q"7"

a.

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I-DO (rag/L) 600 ng K IIL Conductivity I.

II I "-

7.* ;.17

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-, emperamr-e-----L-*L-I[,---tl---*n:l--.

-*mttt-II.---L,-*4 *i-J Loci I'

~j U

DO, (rgC

)

750 migKCl/L Conductivity iil/

poH(S.U).

70 7.

90s0 mg KCILr Conductivity 00...

Temperature

"*.0

%*3

.S %q X

q

~,

p H(S...

),

T*nitissl Final Initial II F*inal iInitial II Final I

Tnitial. II mino1 I

ahE

.t peratures performed at the time of test initiation, renewal or terraination by the analyst identified in the Daily Renewal Informaation table located pn Page 1.

IIknity and hardness performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by:

Page 82 of 95 SOP AT21 - Exhibit AT21.1, revision 11*-01-14

SEnvironmental Testing SoluLtions, Inc.

Ceriodaphnia dubia Chronic Reference Toxicant Control Chart

.1.14 1.12 1.10 1.08 1.06 1.04 1.02 2.5 2.0

~USIEPA ControlI Limits (+/- 2 Standard Deviations)-

I' I

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I EUSEPA Warning and Control Limits (75h and 90t Percentile CVs)

f...

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I I

I u3.6 Test date 7-day IC25 = 25% inhibition concentration. An estlination of the concentration of sodium chloride that would cause a 25% reduction in Ceriodaphnia reproduction for the test population.

.Central Tendency (mean IC25)

Warning Limits (mean IC25 +/- SA.l0 or SA75)

. ~~Control Limits (mean IC25 + SA* SA.90' or" 2 Standard Deviations)

Page 83 of 95 G~llhsgonoroted from assoojatod excel *m*s~ted t.

Extol spradoheet ernerd by: J. Sumner Reevaow by:

w 0

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Ceriodaphnia dubia Chronic Reference Toxicant Control Chart Test number Test date 7-day iC*

CT Is/L Nat'i)

(gIL NaQi) 12 3

4 5

6 7

- 8 9

10 11 12 13 14 15 16" 17 18 19 20 02-04-14 03-04-14 04-08-14 05-08-14 06-03-14 07-08,-14 08-05-14 09-09-14 10-07-14 10-07-14 11-04-14"

" 12-02-14 01-13-15 02-03-2-5 03-03-15 06-07-15 05-05-15 06-09-15.

07-14-15 08-04-15 1.06 1.07 1.05 1.05 1.05 1.07 1.08 1.07 1.08 1.08 1.07 1.08 1.08 1.07 1.06 1.06 1.07

£.08 1.05 1.06 1.08 1.06 1.06 1.06

£.06 L0S 1.06 1.05 1.08 1.105 1.OB 1.05 1.06 1.08

£06 1.05

1.06 1.06

£107

£.07 State and USEEi S

Control Unilts Cr-2S Cr42 0.01

£*04

£.08 0-01 1.04

£.06 a.0 1.04

£.06 0.01 1.04

£.08 O.oi 1.04 1.08 0.01 1.04 1.03 0.01 1.04 1.08 0.01 1.04 1.08 0.01 1.04

. 1.08 0.01 1.04

1.08

.0.01 1.04 1.08 0.01 1.04 1.08 0.01 1.04 1.08 0.01 1.04 1.08 0.01 1.04 1.08 0.01 1.04.

1.08 0.01 1.05 1.08 0.01

£.04 1.08 0.01

£.05 '

1.08 0ao1

£05 1.08 Laboratory Warning Limits Cr-SL*

Cr +5I.

Laboratory SA.*,

Control Limits CT"- Skis CT +SA.S USIPA Sus-warning Uimits CT-S~,*s C1"+S$*.l 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.09 0.08 0.09 0.09 0.09 0.08 0.09 0.09 0.09 0.09 0.09 0.09 0.98 0.88 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.98 0.88 0.88 0.98 0.88 0.98 0.98 0.88 0.08 0.98 0.98 1.14 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 1.15s 1.15 1.15 1.15 1.15 U08PA S==

Control Umits CT. Ski C1"+S5.*

0.1.8 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 0.13 0.18 0.18 0.18 0.18 0.18 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 0.88 0.88 0.88*

0.88 0.88 o.8.8 0.88

£.24 1.24

£.24

£24

£.24

£.24 1.24 1.24 1.24" 1.24 1.24 1.25 1.25 1.25 1.25 GAS8 0.48 0.48 0.48 CAB8 0GAB 0.48 OA48 0.48 0.48 0.48.

0-48 0.48 0.48 0.43 0.48 0.48 0.48 0.48 0.48 0.58 0.58 0.58 0.58 0.58 0.58 0.58 0.58 0.58 0.58 0-58 0.58 0.58 0.59 0.59 0.59 0.50 0.58 0.59 0.59

£.54

£.54 1.54 1.54 1.54.

1.54 1.54 1.54 1.54

£.54 1.54 1.54

£.54 1.54 1.54 1.54 1.54 1.54 1.54 1.55 0.,6 0.66 0.66 0.66 0.66 0-86 0.66 0.66 0.66 0.66 0.86 0.86 0.66 0.66 0.68 0.866 0.66 "0.566 0.66 0.66 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0A40 0.40 0A40 0.40 0.40 0.40 0.40 0.40 1.72 1.72 1.72 1.72 1.72 1.72 1.72

1.72 1.72 1.72 1.72 1.72 1.72 1.72 1.72

£.72 1.72 1.72 1.73 1.73 CV 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.0:1 0.01 0.01 0.01 0.01 Note:

7-dl IC5= = 7-day 25% inhibition concentration. An estimation oft CT = Central tendency (mean IC,5.

S = Standard deviation of the ICzs values.

Laboratory Contbrol and Warning Limits Laboratory control and warning limits were establish ec warning limits recommended by USEPA for the test roe 5*== Standard deviation corresponding to the 5*= Standard deviation corresponding to the ie concentration of sodium chloride that would cause a 25% reduction in Ceriodophrnio reproduction for the test population.

using the standard deviation of the ICs values corresponding to the 10th and 2.5th percentile CVs. These ranges are more stringent than the control and Itrod and endpoint.

1011 percentile CV. (S5* = 0.081 2551 percentile CV. (5S,= 0.17i 75? percentile CV. (5v = 0;45l 9011 percentlie CV. (SA.O = 0.62}

USEPA Control and Warning Limits 5

A.7s5= Standard deviation corresponding to the S4,soQ= Standard deviation c~orresponding to the CV/= Coefficient of variation of the ICes values.

USEPA. 2000. UndarStanding and Actountilg for Method Variability In Whole Effluer tlToxirdty Applications Under the National Pollutant Discharge Elimination Program. EPA-833-R-00-003. US Environmental Protection Agency, Cincinnati, OiH.

Fila: CdNaClOR_080415,xdsx Entered by: J. Sumner Reviewed by:.4m.

"ET

" Environmental Testing Solutions, inc.

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

0 hi

4 e-40 35 30 25 20 15 I

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North Carolina Acceptance Limit (< 40.0%)

Kentucky Acceptance Limit (< 3 0.0%)

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

40 0* 30 S20 10 :

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I U.i Test date

-- e--- 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)

-Page 85 of 95 Graphs gernerted'f rm Rssocfated excel spr*ea*dsheet.

Excel spreadsheet entered by: J. Sumner Reviewed by:

4~Environmental Testing S*olutions, Inc.

Precision of End point Measurements Ceriodaphnia dubia ChronicReference Toxicant Data Testnubr Test date Control Control Mean Survival Reproduction (36)

(offspring/female) 1 2

3 4

5 6

7 8

9 10 11 12 13 14 15 16 17 18 02-04-14 03-04-14 04-08-14 05-06-14 06-03-14 07-08-14 08-05-14 09-09-14 10-07-14 10-07-14 11-04-14 12-02-14 01-13-15 02-03-15 03-03-15 04-07-15 05-05-15 06-09-is ATIAA_

C4_

100 100 100 100 100 100 100 100 100 100*

100 100 100 100 100 100 100 100 Inn/'

30.7 31.3 32.4 31.3 34.6 34.5 34.0 32.6 29.7 32.1 2.9.8 33.1 32.7 32.5 32.4 32.3 31.7 29.7 CT for Control Mean Reproduction (offspring/female) 31.0 31.5 31.4 32.1 S32.5 32.7 32.7 32.3 32.3 32.1 32.2 32.2 32.2

.32.2 32.3 32.2 32.1 2,, A CVa for Control (36)

Reproduction CV(36)

MSD PMSD (36) for PMSD (36) ar 3.5 3.4 6.7 5.2 3.9 3.1 5.0 4.6 4.8 3.1 5.0 6.4 3.8 3.9 6.0 4.6 5.6 2.8

'"FIA 3.4 4..5 4.7 4.5 4-3 4.4 4.4 4.5 4.3 4.4 4.6 4.5 4.5 4.6 4.6 4.6 4.5 1.9 1.8' 2.1 1.7 2.3 1.7 2.3 2.0" 2.2 2.1 1.8 2.0 1.8 1.8 1.9 1.8 1,8 1.4 6.2 5.9 6.5 5.5.

6.6 4.9 6.8 6.1 7.4 6.6 6.2 6.2 5.6 5.6 5.8 5.5 5.8 4.8 6.0

6.2 6.0 6.1 5.9 6.0 6.1 6.2 6.2 6.2 6.2 6.2 6.1 6.1 6.1 6.1 6.0 20 08-04-15 1O00 31.0 32.0 3.0 4.4 1.6 5.0 5.9 Note:

C'V =Coefficient of variation for control reproduction.

Lower CV bound determined by US EPA (10Th percentile) =8.9%.

Upper C'V bound determined by USEPA (90"' percentile) = 42%

MSD =Minimum Significant Difference PMSD = Percent Minimum Significant Difference PMSD is a measure of test precision. The PM~SD is the minimum percent difference between the control and treatment that can be declared statistically slanificant In a whole effluent toxicitv test.

Lower PMSD bound determined by USEPA (10"' percentile)= 13%.

Upper PMSD bound determined by USEPA (g0t' percentile}) = 47%6.

aT = Central Tendancy (Mean Control Reproduction, CV, or PMSD)

USEPA. 2000. Understanding and Accounting for Method Variability in Whoie Effluent ToxicityApplications Under the National Pollutant Discharge Elimination Program. EPA-833-R-V0-003. US Environmental Protection Agency. Cincinnati, OH.

USEPA. 2001a, 200Tb, Final Report: Interiaboratory Variabilit~y Study of EPA Short-tern, Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes l and 2-Appendix. EPA482I-R-01-004 and EPA-821-B-01-O05. US Environmental Protection Agency, Cincinnati, OH.

v, File: CdNaCICROSO841S.xlsx E~ntered by: 3. Sumner P age 86 of 95 Reviewdby:___

T, 0

Page 1 of 6 Sodium Chloride Chronic Reference Toxicant Test (IEPA-821-R-02-013 Method !002.0)

Species: Ceriodaphnia dubia CdNaCICR #: 17_.7 Dilution preparation information-Comments:

NaCI Stoc~k INSqS number:

INSS &

Stock preparation:

100 g NaCIIL:

Dissolve 50 g NaC1 in 500 mL Milli-Q water.

Diuinpe rgL 0

800 1000.

1200 1400 Sto]k o 1

tt e

i)J.~

1 2 1

15 I

18 I

21 Dilent volume ImL

.91 1488 "1485 1482 1479 Totl olme(m). 150

,1500=

1500 1500.

,,1500 STe'st organism source information:.

Test in~formation:

0 ganism.age:

< 24-hours old Randomizing leaplate color I5 I

Date and times organisms were born oL@#rla *t.

'L5 Incubator number and shelf between:

loctiouz t Culture board: J'G'*-21%J A

Replicatenumber:

1 2 I3 l4 I5 6

7 Is 9

10 YoW*

batch:

Cultureboardoupnumrber:

"g '4I*

Ii Ira It. %t VL ?ltl 1t**,$.

Transfer vessel informoation:

pH ='1,

S.U:.. Ternperature = L,

0C Sejenastirw batch:

Ave eragansfer volum e (mL:

0.0121 reL i.A Daily renewal information:

r Day Date Test initiation and feeding, MHSW Analyst renewal and feeding, or batch used termination time 0 ~ ~~08-04-15

~

b,

(

1 AQ.fl~I ~

dtt p

I..

I, I --- E-a'4" -

-I-----,

I-

,.i 08 o-07-15

g5**-s*

4 o08o-15 O&So oA* oq-I S

,i 7

08-11-15 SControl information:

Acceptance criteria ISummar of lest endpoints:1

% of Male Adults:

'<20%

j"7-day LCso

> i'.0,.

% Adults having 3MaBroods:

LOCb1,

>80% "'

NOEC 0

% Mortality:

C)7...

g20%

LOEC j

t)

Mean Offspring/Female:

3(. 0.

>_ 15.0 of'pring/female'

" ChV jq~

t~dlpt~dntan

Page 87 of 95 SOP ATI4I - Exhibit AT14. 1, revision 11-01-14

°aT a

A Page 2of 6 Species: Ceriodaphnia dubia

CONTROT, CdNaCICR #: 17_.7.

Survival and Reoroduction Data

__Replicate number Davy

_1 1

II 7

8 9

1 1

onproduced O ~C)

__I Adult mortality 1

L.

s._.

g.,

2 Yonngproduced I 0I

-6 C)

I_

10 Adu...t mortality o, j

°

° I

3 Young produced 0

0

______IungIKprodu__ed Adult mortality I

.-- q*--

I

[*-'I Ttlyoung pro~dued

'L-

TL b

V0* *-*

_ina A dultMortality I I

I I'-

- I k.

Ioe Adult mortality

'-=

lie

-da) IB sptbro (sigl brodspi b'-Ias)

O=cn oe ofpi g Fiarre ov ihadult Mo rtaity transfer).

w o

%M*orta& ty C1 I.

IMean Offspring/Female:

I *t. 0 I "xI 600 m~NaCI/L Survival and Reproductiorn Data.*

1

______ Replicate number Da-y 1

121314 15161-7 f8 19 1 0 1

Young produced 0 I0 I*

0I Adult mortality L__

L_

L=.

L_

L._

k_.

L...

L_..

a

~

i-I.

2 Young produced 61010 0

0 Ci I OI <3 I Adult mortality

____ TAdult mortality jt.

I I

L.I"'.""

ii'

____3 5

ItL Youngprodueed I

31i, 3~i,4 7

...Irw.

T ougrdu ccmdtlly i

-.1. L

'6

"'i L

,I I '... I ---" [ I. I

'*-

6 Young produced.JC 0

Z 01C O

i t

Adulitmorli,,ty C4

,-s-I'I

\\L_'

-ik

-Ii.

71 Y oungproduced tii I'1

S Iq.

ut,

ib 1*,

t "I I Total young produced s"-

5 3g I 3',

Final Adult Mortality

-w,

-C,,.--,,

i,,.

L i""

,I. -

1,,.,,,,

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

IConcentration:

% Mortali:

flMean Offspring/Female:

, 31.

% Reduction from Control:

"2* io SO P ATl14 - Exhibit A T 14.1, revision I 1-01 -14 88 of 95

4* ~

Page 3 of 6 7 ":

i' i

i 1]

i i

I

.Species: Ceriodaphnia dubia 800 i NaCJ/L CdNaCICR #: 17._f7 Survival and Reproduction Data S~~~~Replicate number Day,,,,___2________7___9 I

Adult mortality

1.

L.

3 Youngproduce IC) of

=

1*l*

C I._aI(

Adultmortlity I

1-I'..*-I.. *.

__ _ Adu.ltortait,

- I Y A~tor~t

_ i*lqI*I,

¥ung produced I

I W1 I

AFWV 6

Y oung produced 1*

(**0 CLO CL _1 lT 0 l 0~

____l Adult Mortality

""1.

L

' II

...Y ug.r.

ue..f I

I ll Note: Adult mortality (L = live, D - dead), SB --split brood (single brood split between two days), CO e arly over (offsring carried over with adult during transfer).

% Mortality 0'..

IMean.0.ff~pring(/Female:

I &A*.

% Reduction from Control:

.7

!1000 mg NaCI/L Survival and Reproduction Data "1

Replicate number 1

Young produced

('3 t*

~ -J ~

4

,~-....

4 Adult mortality

'-~

L_

L.-

L._

LI L..

L..

1... I,..-.

U a~I~I~ee-n~

2 Young produced I C) I (

oicr 0101 CI c%(*Io 0

I I

I.

I. -

1 Y~-

I -~I=--- I

I'-*~--

I.-

~---

i i...;

Adult mortality

-..I

-----I

---I-L...

i i i

.IYoung produced

,O e

'oiZ.

(

I.I Aduit mortalityf

--I "*- I

-TC9-

--,- I.--

Adult mortality !

L,-,

LL.,I-L_

L,-4

__[

Young produced [~z~ J~

.+~

Total young produced

"/-j "2"

3.

I o

21.*

)

j 3- 0 Final Adult Mortality C. CT "L_

i..

_ ~

Note: Adult mortality (L = live, D =dead), SB -*split brood (single brood split between two days), CO = carry over (offpring carried over with adult during transftr).

IConcentration:

% Mortality 07 iiMean Offspring/!Female:

Iq S% Reduction from Control: I

.2.o SOP AT14 - Exhibit AT14.1, revision 1 1-01-14 89 of 95

Species: Ceriodaphnia dubia 1200 m~NaCI/t Page 4of6 CdNaCICR #: 17._7 Survival and Reproduction Data I

___Replicate number Day

_I 2

3*

4 5

6 7

S 9

10 1

[

oung poduced 0

"0 010

__}Adult mortality Young produced

[

(

C){

I I -*.

A Adult mortality j[~=='-

3If Young produced 1 1..~....

..~j0 4if Youugproduced J 15

]

I.'--~I

' -- T

'¢ ]

I

,*l SAdult mortality J,

\\...1\\..

---L*-

I I_- 1

'*-Il*-

5 Youngproduced

- ~l 11 l

1+/-l-e -

J-11 ' __- -' I _

To6 youngproduced

[

0

.A*,.

X f0

]O

___a AdultrMo rta lit, i -

I I

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

Concenration:~

% Mortality:

AMean Offspring/Female:

I

% Reduction from Control:

12.5...,3***

1400mg NCI/LSurvival and Reproduction Data

_.__Relca ume 1

Young producedd,I

It%

I*

O I 0 I 0 I 0*0*0 I*II l

l I It I

I 1

~-l

~

i1 I ______

i

.~

.L -~-

I -~-

2 Young produced ci oI 01 el cIo*l0I 0 r*

Adult mortllty oungpr--oId--Ie---I

--l*

.* I___ I__

l_

L

Adult mortality

__i.-

' I_

4 Yousng produced jC j0 0

.I)

C

Q.Q f

ii Adult mortality

-__7

'-*.-I

'- 1_ -

i Young produced

...... I*20 0

2

.I I

__ -Adult mortalityj LjL_[

C.J L

'..Is.

Yngproduced -__

I0ji i

.. jf I

Adult

__ortality~.

~'C Total young produced

.I...

~

1 inlAdult Mortalit C

C

-c I Note: Adult mortality (L =live, D = dead). SB =split brood (single brood split between two days), CO c arry over (otispring carried over with adult during transfer).__________________

Concentration:"

I% Mortality: *(*

Mean Offspring/Female:

"2 II% Reduction from Control: I 2T..q 7 SOP AT14 - Exhibit ATI 4.1, revision 11 14

0 A

1t Environmenltal Testilng Solutions, Inc.

0ii Control

Ve rification of Ceriodaphnia Reproduction Totals

Day 1

2 Re3caenmbr4 Total 1I 0

0 0

0 1

0 2

0 0

0 000 01 0

0

-3 0

0 0 0o 0

0 4

4 3

4 4S4 34 4

4 39 5

12 12 13 101 11 1

1 1

3 118 6

0 0 0 o

0 0

7 16 I5 15 171 15 14 17 15 1

153 Total 32 30 32 31 31 3I0 3

0 3

310 D y R e *lica e num ber Total 1a 2-3 I4 5

6 7

8 9

1

.I "0 -

1 0

0

0.

0 0

0.0 0

0 0

2 0 ' 0 0"

0 0

_0 00 0

3 0

00O0 0

0 0

0 00 0

5 2

12 13 12 13

.11 11 121 3

1 120 6

0, 0'

0 0

0 o

0 01 0

o 7

17 17 15 16 14 i1615 1511 1

157 Totl 34, 33 32 33 30 32 30 31 3

30 319 800 mg NaCIJL--

Day

~~~Re l1icate number Toa Da ]2 3

4 5

6 7_

I 9ot10 1

0 0

0 0 0

0 0

0 2

0I 0 o

0l 0 0

0 0o 0

3 0

0 0

0 0

4 5

4 4

3 4

54 4

4 41 5

12 12 12 11I 10 13 I12 10 2

12 116 6

0 0

0i 0

0 0 0 0

0 7

16 11 14, 14 I15 14 17 151 15 152 Total 33 33 30 28I 29 32 "33 29 1

31 309 1000 mg NaCi/L Re liicate number Day 1

2 3

4 5

6 7

,, 8 9

10 Total 1

0 0

0,0J 2

0 0

0

3.

0 0

0 4

4 3

4 4

5 4

5 5

4 4

42 5

10 10 11

.13 10 12 13 10 10 12 111 6

0 0

0 00..0 0

0 a

7 14 14 16 13 16 14 12 15 13 14 141 Total 28 27 31 30 31 30 30 30 27 30 294 1200 mgNaCI/L DyReplicate nuinber

-~Total Da i

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 4

3 5

3 2

4 2

3 3

3 2

30 6

0 0

0 *10*

0 0

23 7

2' 2

5 6

0 4

7 0

0 5

31 Tota 10 11 10 8

7 6

10 6

9J7 84' 1400 mg NaCiLL Replicate numberToa Day 1

2 3

4 5

6 7.*8 9

Tota 1

0 0

0 2

0 0

0 0"

0 0

0 3

0 0

0 4

0 0

0 2

0 0

1 0

0 0

3 5

0 0

0 3

5 6

0 0 '.0 0

0 0

0

.0 0

7 2

1 1

0 0

3 1

1 2

3 14 Total 2

1 1

2 2

3 2

1 5

3]

22 File: CdNaClCR_080415.xlsx Entered by: J, Sumner Reviewed by:

1%)

0~

Environmental Testing Solutions, Inc.

CerJ odaphnia dubia Chronic Reference Toxicant Test EPA-821-R-02-013, Method 1002.O Verificati Quality Control 3fn of Data Entry, Calculations, and Statistical Analyses Test number:

Test dates:

CdNaCICR #177 Auut 41121 Concentration Repli rate number

_Survival Average reproduction Coefficient of Percent reduction from (mg/L NaCI)

I 2

3 4

56 7

8 9

10

(%)

(offsprlng/female) variaton (%)

control ('A)

-*i I

Control 32 30 32 31 31 30 30 32 30 32 100 31.0 3.0 Not applicahle 600 34 33 32 33 30 32 30

-31 34 30 100 31.9 5.0

-2.9 800 33 33 30 28 29 32 33 29 31 31 100 30.9 6.0 0.3 1000 28 27 31 30 31 30 30 30 27 30 100 29.4 5.1 5.2 1200 io it t0 a

6 10 6

9 7

100 8.4 21.9 72.9 1400.

2 1

1 2

2 3

2 1

5 3

100 2.2 55.9 92.9 Dunnett's MSD value:

PMSD:

1.563 5.0 MSD Miim PMSD =

Perceni PMSD signific Lower:

Upper:

Lower.

Variabl USEPA. 2001a, 200lb. Final Report: Interlabo story Variability Study of EPA US Environmental Protection Agency, Cincinnati, OH-.

Minmm Significant Difference is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically ant in a whole effluent toxicity test.

'MSD bound determined by USEPA (10t? percentile) =13%.

'MSD bound determined by USEPA (90lh percentile) = 47%.

md upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET Interlaboratory lity Study (USIEPA, 200 [ a; USEPA, 200I b).

Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes 1 and 2-Appendix. EPA-821-B-01-004 and EPA-821-B3-01-005.

File: CdNaCICR_080415.xlsx Table populated from associated "Verification of Cerdodaphnia Reproduction Totals" spreadsheet Spreadsheet entered by: J. Sumner

~Reviewed by::_Af_.

0 0 F iO lIItlb flSI5 bI f.

Statistical Analyses Ceriodaphnla Survival and Re0roduction Test-Reproduction

.Start Date:

8/412016 End Date:

8/11112015 Sample Date:

Comments:

Conc-mgIll 1

2 D.Control 32,000 30.000 600 34.000 33.000 800 33.000 33.000 1000 28.000 27.000 1200 10.000 11.000 1400 2.000 1.000 Test ID:

CdNaCICR Sample ID:

REF-Ret Toxicant Lab iD:

ETS-Envlr. Testing Sol.

Sample Type:

NACL-Sodlum chlioride Protocol: FWCHR.EPA-821.R-02-013 Test Species:

CD-Ceriodsphnla dubia 3

4 5

6 7

8 9

10 32.000 31.000 31.000 30.000 30.000 32.000 30.000 32.000 32.000 33.000 30.000 32.000 30.000 31.000 34.000 30.000 30.000 26.000 29.000 32.000 33.000 29.000 31.000 31.000 31.000 30.000 31.000 30.000 30.000 30.000 27.000 30.000 10.000 8,000 7.000 6.000 10.000 6.000 9.000 7.000 1.000 2.000 2.000 3.000 2.000 1.000 8.000 3.000 Transform: Untransformed Conc-mnI1L Mean N-Mean Mean Mmn Max GV%

N D-Contrcl 3t.000 1.0000 31.000 30.000 32.000 3.041 10 600 31,900 1.0290 31.900 30.000 34.000 5.000 10 800 30.900 0.9968 30.900 28.000 33.000 5.997 10 "1000 29.400 0.9484 29.400 27.000 31.000 5.121 10 "1200 8.400.

0.2710 8.400 6.000 11.000 21.679 10 "1400 2.200 0.0710 2.200 1.000 5.000 55.876 10 1 -Tailed Isotonic t-.Stat Critical MSD Mean N-Mean 31.450 1.0O000

-1.316 2.287 1.563 31.450 1.0000 0.146 2.287 1.563 30.900 0.9825 2.340 2.287 1.563 29.400 0.9348 33.057 2.287 1.563 8.400 0.287t 42.125 2.287 1.563" 2.200 0.0700

.Auxiliary Tests Statistic Critical Skew Kurt Kolmogorov D Teat indicates normal distribution (p > 0.013 0.85666 1.035

-0.0915 -0.9489 Bartlett's Test indicates equal variances (p~ = 0.40) 5.13552 15.0863 Hypothesis Test (t-tail. 0.05)

NOEC LOEC CliV TU MSDu MSDp MSB MSE F-Prob df Dunnett'sTest 800 "1000 894.427 1.58333 0.05043 1778.588 2.33704 0.OE+00 5, 54 Treatments vs D-Control.

Point IC05 IC10 IC1 5 1C20 1C25 IC40 iC50 Unear Interpolation (200 Resamples) mg/L SD 95% CL Skew 936.333 69.1013 771.977 1003.81

-1.1921 1010.43 5.81691 1002.55 1016.58

-3.2768 1025.4 4.06401 1017.88 1033.34

-0.2997.

1.0]-

1040.38 3.87909 1033.32 1046.14

-0.3096 0,9]

1055.36 3.75027 1048.63 1062.99

.0.3195 j.8 1100.29-3.74598 1093.57 1107.3

-0.32110.

1130.24 4.05478 1122.81 1137.99

-0.28085 0.7-.

0.6-0.

l" 0.3-0.2-0.1t Dose.Response Plot-0.1~~

0 500 1000 1600 Dose mgIL Dose-Response Plot 1-tail. 0.05 levei of significance File.: CdNaCICR..HO4l5.xlsx Entered by: 3. Stunner Itaviewed by:__

93 of 95

ET Eng~ IrO~n Page 5 of 6 Species: Ceriodaphnia dubia CdNaCICR #: 1771 1)ipChemistry Day I(Analys identified fbr each d, performed pH, D.O. and cond~uctivi* measurements oniy)

A II I

2 4

Analyst I #/]/4 Concentration Parameter pH (S.u.)

DO (rag/L)

Conductivity (usnhoslcm)

CONTROL

.4

Aflalinity S*Hardness I(rag CaCOv'L)

'Temperature (°C) pH (s.u.)

7?.q II17.20:E I7.#I

-zIii-IJ~u I600 mg NaCI/L DO (mg/L)

II -,,

[Conductivity I~L~

'Teminerature (°C) pH (SU.})

' 800 mg NaCiIL Con-i etivl I

"*Temperature (-

I "C).

I I *%s lY pH (S.U.)

I 2--

!1 nfl

~

-~ I)

-9 I~

I

/*i *,,,t,

--F

I -U

'IiJU /'j-J i11II

¢1 1000 mug NaCI/L Cond ctivIIy-

no

Temperature (GC)

II 1AA

oIII 6

6 I --

I*

f pH (s.U.)

7.S*

~qj i~

El DO (raglL) 1200 mg NaCI/L

.5 JI~~.s Conductivity I ~fl7'~

j~

(tzmhoslcmn) vv'-

Temlperature (°C) 7A4~

")..*.*I, II t"Jh* 2

__1 __ _ _

9 I

II~4

~T'~I~

pH (S.U,)

7,4 0

-L,' I I7.

I

1 l

" " I --

II - *-* 2

_ r -*

fl.----

DO (ingle) 1400 mg NaCIIL rConductivity npmos/cm)

"1_ °O LZkJLI

~

Temnerature (oC)

-lii. ~

I

,I Initial I

J~ial

Tem~peratures performed at the time oftest 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, analysis and transcribed to this bench sheet by:

L*

ST

'0 ~l~mftfb~g*bl, Page 6 of 6 Species: Ceriodaphnia dubia CdNaCICR #: 177 Day Anltietified for each da, erformed H,'.. an odcii maureet n

3 4__._5

... _6 DO (mg/)

,v CONTROL

Alkainmity

{L-*

600 mg NaCI/L Conductivity

~

,j

Temnperature A

s.

~

pH (.U.)

__-"-___-" _'---'l**

~

l*-.

800 mug NaCI/L Codciv0°t-'*

'*,zo8' I

-t Q

_ ~(i ic so) f...

pH (S.U.)

II *.*4 I! "*,*

,I ":J.*2 II *.*

!,1a3 II'.*

I DO (ma/Lt a*

-*--*-i!*

41:

1000 mg NaCI/L Conductivity (jxmhs/cm) 7..* 0 "a.*.O I ~

J 7-*

..J

Ternperature (0C)

S1

.I 1K Z2-1 II II.

~

'ii ~4 pH (s.u.)

U *,*

I "4*k I q*

I "-1'iZ DO (rag/b) 1200 ing-NaCI/L Conductivity L$ ~O

.6 1~W~

-' t IL oHL-tI~q

.Temperature (OQ).*

0-

-II h-

.t I

I 1

I-

, -l11*

DO.(mg/)

_i~

-*--]

Initial Final initial lI Final Initial Final initial Final bTemperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Intfomaafion table located on Page 1. Alkalinity and hardness performed by the analyst identified on thQ bench sheet specific for each

p

~analysis and transcribed to this bench sheet by:

4 a~

o[95 SOP AT14 - Exhibit ATl4. 1, revision 11-01-14

v t e Letter Sequence Request
TAC:MF5366, (Approved, Closed)
Initiation Request, Request Acceptance... Supplement Administration Questions Answers 11 September 2015 Results Approval Other:
MONTHYEAR2015-10-23 [Table View]

ML15258A023

Text

Dear Ms. Morgan:

SUMMARY

Contact:

Contact:

Contact:

SUMMARY

SUMMARY

SUMMARY

SUMMARY

SUMMARY

'*-

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