Ms. Christina Morgan Division of Water Pollution Control ... · Ms. Christina Morgan Tennessee...

Tennessee Valley Authority, Post Office Box 2000, Soddy Daisy, Tennessee 37384-2000

October 10, 2012

Ms. Christina MorganTennessee Department of Environment

and ConservationDivision of Water Pollution ControlEnforcement & Compliance Section6 th Floor, L & C Annex401 Church StreetNashville, Tennessee 37219

Dear Ms. Morgan:

TENNESSEE VALLEY AUTHORITY (TVA) - SEQUOYAH NUCLEAR PLANT (SQN) - NPDESPERMIT NO. TN0026450 - DISCHARGE MONITORING REPORT (DMR) FOR SEPTEMBER 20"12AND REVISED DMR FOR AUGUST 2012

Enclosed is the September 2012 Discharge Monitoring Report for Sequoyah Nuclear Plant. Thetoxicity report and revision to Discharge Number 101T, Biomonitoring for Outfall 101 for August2012, are also attached. There were no exceedances during the monitoring period. If you have anyquestions or need additional information, please contact Brad Love by email at [email protected] orby phone at (423) 843-6714.

I certify under penalty of law that this document and all attachments were prepared under mydirection or supervision in accordance with a system designed to assure that qualified personnelproperly gather and evaluate the information submitted. Based on my inquiry of the person orpersons 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. Iam aware that there are significant penalties for submitting false information, including the possibilityof fine and imprisonment for knowing violations.

Sequoyah Nuclear Plant

Enclosurescc (Enclosures):

Chattanooga Environmental Field Office U.S. Nuclear Regulatory CommissionDivision of Water Pollution Control Attn: Document Control DeskState Office Building, Suite 550 Washington, DC 20555540 McCallie AvenueChattanooga, Tennessee 37402-2013

PERMIIrEE NAME/ADDRESS (Include Facility Name/Location if Differentj-

Name TVA - SEQUOYAH NUCLEAR PLANT

Address P.O. BOX 2000- I[NTEROFFICE OPS-5N-SQ.N

SODDY - DAISY, TN 37384Facilj.:, TVA - SEQUOYAH NUCLEAR PLANTLocation HAMILTON COUNTY

ATTN: Brad Love

NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJORDISCHARGE MONITORING REPORT (DMR)

(SUBR 01)

TN0026450 t101 G F-FINAL

PERMIT BEDISCHARGE NUMBE DIFFUSER DISCHARGE

MONITORING PERIOD EFFLUENT

Form Approved.

OMB No. 2040-0004

F r YARm I Mr DAY I T EAo l MO I DAYFrom 1 2-- 0 0 To 1 12 109 1301 NO DIS iSCHARGE

NOTE: Read instructions before completing this form.

PARAMETER FREQUENCY SAMPLEOF

TYPEQUANTITY OR LOADING QUALITY OR CONCENTRATION NO.EX

FREQUENCYOF

ANALYSIS

SAMPLETYPE

AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS

TEMPERATURE, WATER DEG. SAMPLE ** 41.3 0 30/30 RCORDRCENTIGRADE MEASUREMENT

04

00010 1 0 PERMIT *** Req. Mon. DEG. C. CONTI CALCTD

EFFLUENT GROSS REQUIREMENT DAILY MAX NUOUS

TEMPERATURE, WATER DEG. SAMPLE 29.1 04 0 30/30 MODELDCENTIGRADE MEASUREMENT

00010 Z 0 PERMIT 30.5 DEG. C. CONTI CALCTD

INSTREAM MONITORING REQUIREMENT DAILY MX NUOUS

TEMP. DIFF. BETWEEN SAMP. & SAMPLE 3 04 0 30/30 CALCTDUPSTRM DEG.C MEASUREMENT

00016 1 S PERMIT -**- 3 DEG. C. CONTI CALCTD

EFFLUENT GROSS REQUIREMENT DAILY MX NUOUS

FLOW, IN CONDUIT OR THRU SAMPLE 1761 03* * 0 30/30 RCORDRTREATMENT PLANT MEASUREMENT 03

50050 1 0 PERMIT Req. Mon. MGD CONTI RCORDR

EFFLUENT GROSS REQUIREMENT DAILY MAX NUOUSCHLORINE, TOTAL RESIDUAL SAMPLE * 0.017 0.032 19 0 24 / 30 GRAB

MEASUREMENT

50060 1 0 PERMIT 0.1 0.1 MGIL FIVE PER CALCTD

EFFLUENT GROSS MO AVG DAILY MAX WEEKTEMPERATURE - C, RATE OF SAMPLE 0 62 0 30)30 CALCTDCHANGE MEASUREMENT

82234 1 0 PERMIT * 2 DEG **wewe** ***,,* CONTI CALCTD

EFFLUENT GROSS REQUIREMENT DAILY MX CIHR NUOUSSAMPLE

MEASUREMENT

PERMIT

REQUIREMENT

NAM1_J'rITILE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my TELEPHONE DATEdirection or supervision in accordance with a system designed to assure that qualified personnel

John T. Cadin properly gather and evaluate the information submitted. Based on my inquiry of the person or [ Pepersons who manage the system, or those persons directly responsible for gathering the 423 8i-7 President i ninformation, the information submitted is, to the best of my knowledge and belief, true, accurate. 423 843-7011 1 n

- .and complete. I am aware that there are significant penalties for submitting false information, SI E OF PRINCIPAL EXECUTIVE

including the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER MO DAYTYPED OR PRINTED I CODE

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

No closed mode operation. Veliger monitoring data is included as an attachment. The following injections occurred: 1. Floguard MS6236 (max. calc. conc. was 0.057mg/L-limit 0.2mgIL) 2.Biodetergent 73551 (max. calc. conc. was 0.038mg/L-limit 2.Omg/L) 3. Spectrus CT1300 (max. calc. conc. was 0.037mg/L-Iimit 0.050mg/L)

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

Mean # of Water Mean# of Water NOTES: %Sample Date % Settlers Sample Date Asiatic Temp. ('C) LOCATION SUB LOCATION Gravid Asiatic COLLECTED BYCClamsm3 TClam

01/03/2012 14 100 26 01/03/2012 0 26 1-25-545 PKS01/10/201201/17/201101/24/201201/31/201202/07/201202/14/201202/21/201202/28/201203/06/201203/13/201203/20/201203/27/201204/03/201204/10/201204/18/2012May 2012June 2012July 2012

August 2012September 2012

00000000000

162322979

326

00000000000

1.30

205

91013

17.6128.3

26.511.111.713

14.617.21822

18.8

01/10/201101/17/201101/24/201201/31/201202/07/201202/14/201202/21/201202/28/201103/06/201203/13/201203/20/201203/27/201204/03/201204/10/201204/18/2012

000000000000000

91013

17.6128.3

26.511.111.713

14.617.21822

18.8

RCW1-ISV-24-1234

1-25-5451-25-5451-25-5451-24-12341-25-545

1-ISV-24-12341-ISV-24-12341-ISV-24-12341-ISV-24-12341-ISV-24-12341-ISV-24-12341 -ISV-24-12341-1SV-24-1234

WBEPB

WDTCRBB

WECR

WBEWBEWBEWBEWBE

PBPB

MJWNo Samples CollectedNo Samples CollectedNo Samples CollectedNo Samples CollectedNo Samples Collected

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

N±ame TVA - SEQUOYAH NUCLEAR PLANT

Address P.O. BOX 2000-_ -_ (INTEROFCE OPS-5N-SQN.)

SODDY - DAISY TN.37384Facil_ y_ A - SEQUOYAH NUCLEAR PLANT

LoCation HAMILTON COUNTY

ATTN: Brad Love

NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR ForDISCHARGE MONITORING REPORT (DMR) (SUBR 01) OM

TN0026450 F-FINAL

PERMIT NUMBER DISCHARGE NUMBER BIOMONITORING FOR OUTFALL 101

MONITORING PERIOD EFFLUENT

rm Approved.

BI No. 2040-0004

Froml 1 109011 Tol11 09L L 30 NO DISCHARGE liZ ]NOTE: Read instructions before completing this form.

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

AVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS

IC25 STATRE 7DAY CHR SAMPLE MonitoringCERIODAPHNIA MEASUREMENT Not-Required 23

TRP3B 1 0 PERMIT *"***** eee*** 43.2 PERCENT SEMI COMPOS

EFFLUENT GROSS REQUIREMENT MINIMUM ANNUAL

IC25 STATRE 7DAY CHR SAMPLE ** Monitoring 23PIMEPHALES MEASUREMENT Not RequiredTRP6C 1 0 PERMIT 43.2 * PERCENT SEMI COMPOS

EFFLUENT GROSS REQUIREMENT MIMINUM ANNUAL

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMIT

REQUIREMENT

NAMEFTITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my . •TELEPHONE DATEdirection or superyision in accordance with a system designed to assure that qualified personnel

John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the.person orpersons who manage the system, or those persons directly responsible for gathering the President 43 8.-0! tinfonnation the information submitted is, to the best of my knowledge and belief, true, accurate. .. 4.3

land complete. I am aware that there are significant penalties for submitting false information, SIGhTJp:F PRINCIPAL EXECUTIVET including the possibility of fine and imprisonment for knowing violations. OFFICeR'OR AUTHORIZED AGENT AREA NUMBER MO DAY

__"c'_____________AUTHORIZEDAGET________R YEEARJ OIDýITYPED OR PRINTED I CODE


Toxicity was not sampled in September 2012.

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

PERMITTEE NAME/ADDRESS (Include Facility Name/Location if Different)Name TVA - SEQUOYAH NUCLEAR PLANTAddress P.O. BOX 2000

.f.-.NTEROFFICE OPS-5N-SQON)SODDY -DAISY. TN 37384

Facilityy TVA - SEQUOYAH NUCLEAR PLANTLocation HAMILTON COUNTY

ATTN: Brad Love

NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES) MAJOR Form Approved.DISCHARGE MONITORING REPORT (DMR) (SUBR 01) 0MB No. 2040-0004

TN024 1103 G F-FINAL

PERMIT NUMBER DISCHARGE NUMBER LOW VOL. WASTE TREATMENT POND

m•IIIITnRINA PFRI"nn I EFFLUENT

I YEAR Mo "D oEAR I DAY

From[ 12 0 I1 I To 1 12 i 09 30 NO DISCHARGE [ *] *

NOTE: Read instructions before completing this form.PARAMETER QUANTITY OR LOADING I QUALITY OR CONCENTRATION NO. FREQUENCY SAMPLE

SEX OF TYPE


PH SAMPLE * 7 9 12 0 19/30 GRABMEASUREMENT

00400 1 0 PERMIT 6 .*9 SU THREE/ GRAB

EFFLUENT GROSS REQUIREMENT MINIMUM MAXIMUM WEEK

SOLIDS, TOTAL SUSPENDED . SAMPLE '9 14 19 0 2/30 GRABMEASUREMENT

00530 1 0 PERMIT 30 100 MGIL TWICE/ GRAB

EFFLUENT GROSS REQUIREMENT MO AVG DAILY MX T MONTH

OIL AND GREASE SAMPLE <1 <1 19 0 2/30 GRABMEASUREMENT

00556 1 0 PERMIT 15 20 MG/L TWICE) GRAB

EFFLUENT GROSS REQUIREMENT MO AVG 'DAILY MX MONTH

FLOW, IN CONDUIT OR THRU SAMPLE 0.948 1.134 0 30)30 RcORDRTREATMENT PLANT MEASUREMENT

03

50050 1 0 PERMIT Req. Mon. Req. Mon MGD. SEE RCORDREFFLUENT GROSS REQUIREMENT MO AVG DAILY MXEFFLUENTAIGROSS PERMIT.

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my TELEPHONE DATElirction or supervision in accordance with a system designed to assure that qualified personnel

John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the person orpersons who manage the system, or those persons directly responsible for gathering the President

Q Ok sit. •;ic P•ree;,de information, the information submitted is, to the best of my knowledge and belief, true, accurate, a 423 843-7001 12 10 10S . ." and complete. I am aware that there are significant penalties for submitting false information, SIA E PRINCIPAL EXECUTIVE

including the possibility of fine and imprisonment for knowing violations. SI ICE -OR AUTHORIZED AGENT AREA NUMBER -TYPED OR PRINTED I CODE

COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all aftachments here)

EPA Form, 3320-1 (REV 3I9S) Previous editions may be used Page 1 of 1EPA Foffn 3320-1 (REV 3199) Previous editions may be used Page 1 of 1

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

Name TVA - SEQUOYAH NUCLEAR PLANTAddress P.O. BOX 2000

S•- J[NTEROFFICE OPS-5N-SQN)SODDY - DAISY TN _37384___..

Fai _•,( TA - SEQUOYAH NUCLEAR PLANTLocation HAMILTON COUNTY

ATTN: Brad Love

NATIONAL POLLUTANT DISCHARGE ELIMINATION. SYSTEM (NPDES) MAJORDISCHARGE MONITORING REPORT (DMR)

(SUBR 01)

TN0026450 110 G F - FINAL

PERMIT NUMBER DISCHARGE NUMBER RECYCLED COOLING WATER

MONITORING PERIOD • I EFFLUENT

Form Approved.OMB No. 2040-0004

I YEAR MO I DA YEAR IDAY NO DISCHARGE F]From1 12 09L101 Tol 1[ 1j09 1j301 -- NOTE: DCHRGe i

NOTE: Read inlstrJctionls before completing this form.



TEMPERATURE, WATER DEG. SAMPLE 04CENTIGRADE MEASUREMENT 04

00010 1 0 PERMIT * ****** REPORT DEG C CONTIN CALCTD

EFFLUENT GROSS VALUE REQUIREMENT. DAILY MX UOUS

TEMPERATURE, WATER DEG. SAMPLE 04CENTIGRADE MEASUREMENT 04

00010 Z 0 PERMIT * 30.5 DEG C CONTIN CALCTD

INSTREAM MONITORING REQUIREMENT DAILY MX UOUS

TEMP. DIFF. BETWEEN SAMP. & SAMPLE 04UPSTRM DEG.C MEASUREMENT - 04

00016 1 0 PERMIT 5 DEG C CONTIN CALCTD

EFFLUENT GROSS VALUE REQUIREMENT DAILY MX UOUS

FLOW, IN CONDUIT OR THRU SAMPLE 03TREATMENT PLANT MEASUREMENT 03

50050 1 0 PERMIT Req. Mon. MGD * * * CONTIN RCORDR

EFFLUENT GROSS VALUE REQUIREMENTDAILY MXCHLORINE, TOTAL RESIDUAL SAMPLE 19

MEASUREMENT

50060 1 0 PERMIT *0.1 0.1 MOIL Five per CALCTD

EFFLUENT GROSS VALUE REQUIREMENT -MO AVG DAILY MX Week

TEMPERATURE - C, RATE OF SAMPLECHANGE MEASUREMENT 04

82234 1 0 PERMIT 2 DEG C *********. ** CONTIN CALCTD

EFFLUENT GROSS VALUE REQUIREMENT DAILY MX uousSAMPLE

MEASUREMENT

PERMITREQUIREMENT

NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my I/ TELEPHONE DATEdlirection or supervision in accordance with a system designed to assure that qualified personne

John T. Carlin property gather and evaluate the information submitted. Based on my inquiry of the person orpersons who manage the system, or those persons directly responsible for gathering the . Vice President

Seiuovah Site Vice President nformation, the information submitted is , to the best of my knowledge and belief, true, accurate, E_ 423 843-7001 12 PN PAnand amp;.; awaw .... iht mhere are significarn penalties tor submitting false information, S-I--- E OFPRNCPA -EECUTIVE

including the possibility of fine and imprisonment for knowing violations. O OR AUTHORIZED AGENT AREA NUMBER

TY OR PRINTED I CODECOMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

No Discharge this Period

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

PERMIIrEE NAMEIADDRESS (Include Facility Name/Location if Diffement)

Name TVA - SEQUOYAH NUCLEAR PLANT

Address P.O. BOX 2000--- LNTEROFFICE OPS-5N-SQ_.N)

SODDY - DAISY. TN 37384

Faciljiy TVA - SEQUOYAH NUCLEAR PLANT

Location HAMILTON co UNTY.-

ATTN: Brad Love

NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM (NPDES)

DISCHARGE MONITORING REPORT (DMR)

TN0026450 11PERMIT NUMBER DISCHARGE R

MONITORING PERIODYEAR MO I DAY I YEAR MO DAY

From J12 09 101 Tol 121091301

MAJOR

(SUBR 01)

F - FINAL

RECYCLED COOLING WATER

EFFLUENT

Form Approved.

OMB No. 2040-0004

** NO DISCHARGE [ý_ *]


PARAMETER QUANTITY OR LOADING I QUALITY OR CONCENTRATION NO. FREQUENCY SAMPLEI __ OF TYPEAVERAGE MAXIMUM UNITS MINIMUM AVERAGE MAXIMUM UNITS ANALYSIS

IC25 STATRE 7DAY CHR SAMPLE

CERIODAPHNIA MEASUREMENT 23

TRP3B 1 0 0 PERMIT *******a 43.2 * PERCENT SEMI COMPOSREQUIREMENT

EFFLUENT GROSS VALUE MINIMUM ANNUAL

IC25 STATRE 7DAY CHR SAMPLE 23

PIMEPHALES MEASUREMENT 23

TRP6C 1 0 0 PERMIT *** 43.2 * PERCENT SEMI COMPOS

EFFLUENT GROSS VALUE REQUIREMENT MINIMUM ANNUAL

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMIT

REQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

NAME/TITLE PRINCIPAL EXECUTIVE OFFICER. I Certify under penalty of law that this document and all attachments were prepared under my TELEPHONE DATEdirection or supervision.in accordance with a system designed to assure that qualified personnel

John T. Carlin property gather and evaluate the information submitted. Based on my inquiry of the person orpersons who manage the system, or those persons directly responsible for gathering the 7--sqo l•a" IteVice PresidentS q o a SieVc Prsd n information, the information submitted is, to the best of my knowiledge and belief, true, accurate, LL "F J.. .. .423 843-7001 12 10 10

Sequoyah Site Vice President and complete. hem a•re tha there r" . , , .fnSREoh n t to d , ee s41includin th os --_.. , . ....... Siu lR ..... PRN IA EX CU IV Iu41t, (• II•~

including the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER YEARMO DAY

TYPED OR PRINTED CODECOMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)

No Discharge this Period

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

PERMI-rEE NAME/ADDRESS (Include Facility NamelLocation if Different)Name TVA - SEQUOYAH NUCLEAR PLANTAddress P.O. BOX 2000

- - - (NTEROFFICE OPS-5N-SQN).SODDY - DAISY TN 37384

Facilijy TVA - SEQUOYAH NUCLEAR PLANTLocation HAMILT.ON COUNTY

ATTN: Brad Love


DISCHARGE MONITORING REPORT (DMRJ

TNO026450 118 GPERMIT NUMBER DISCHARGE NUMBERJ

MONITORING PERIODSYEAR I MO I DAY T o A M I PAYFrom 12T og Ioi I To l 12 109 1301

MAJOR Form Approved.

(SUBR 01) OMB No. 2040-0

F - FINAL

WASTEWATER & STORM WATER

EFFLUENT

NO DISCHARGE [ "*


004



OXYGEN, DISSOLVED (DO) SAMPLEMEASUREMENT **19

00300 1 0 PERMIT 2 MGIL TWICE/ GRAB

EFFLUENT GROSS REQUIREMENT MINIMUM WEEK

SOLIDS, TOTAL SUSPENDED SAMPLEMEASUREMENT 19

00530 1 0 PERMIT 100 MGIL TWICE/ GRAB

EFFLUENT GROSS REQUIREMENT DAILY MX WEEK

SOLIDS, SETTLEABLE SAMPLEMEASUREMENT 25

00545 1 0 PERMIT * * MUL ONCE/ GRAB

EFFLUENT GROSS REQUIREMENT DAILY MX MONTH

FLOW, IN CONDUIT OR THRU SAMPLE 03TREATMENT PLANT MEASUREMENT 03

50050 1 0 PERMIT Req. Mon. Req. Mon. MGD ."*ONCE/ ESTIMA

EFFLUENT GROSS REQUIREMENT MO AVG DAILY MX BATCHSAMPLE

MEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

NAME/TITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my 1 TELEPHONE DATEdirection or supervision in accordance with a system designed to assure that qualified personnel

John T. Carlin properly gather and evaluate the information submitted. Based on my inquiry of the person orpersons who manage the system, or those persons directly responsible for gathering the sident . -information, the information submitted is, to the best of my knowledge and belief, true, accurate. AA-7flI , u ,IUStVcPein n-_ C'..!cm. ziwm t', ;ha-i atu signiiicant penalties for submitting false information, sIGNA'r-RE OF PRINCIPAL EXECUTIVE /including the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT A NUMBER YEAR MO DAY

TYPED OR PRINTED COCOMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all aftachments here)

During this reporting period, there has been no flow from the Dredge Pond other than that resulting from rainfall.

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

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

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

-JLNTEROFFCE OPS-5N-SQN)

SODDY - DAISY, TN_373_84Fac.lli' TVA - SEQUOYAH NUCLEAR PLANT

L _ocr HAMILTON COUNTY

ATTN: Brad Love


DISCHARGE MONITORING REPORT (DMRJ

TNO02645010TF PERMIT NUMBýER PDISCHARGEI N'UMBER]

MONITORING PERIODýYEAýMO DAY L RMFroml 12108 101 1 To 1 12 1 08 1 31

MAJOR Form Approved.

(SUBR 01) OMB No. 2040-0

F - FINAL

BIOMONITORING FOR OUTFALL 101

EFFLUENT

NO DISCHARGE [LI]NOTE: Read Instructions before completing this form.

004



IC25 STATRE 7DAY CHR SAMPLE >100.0 23 0 1I180 COMPOSCERIODAPHNIA MEASUREMENT 23

TRP3B 1 0 PERMIT *e****** -- ** 43.2 * PERCENT SEMI COMPOS

EFFLUENT GROSS REQUIREMENT MINIMUM ANNUAL

IC25 STATRE 7DAY CHR SAMPLE >100.0 23 0 1/180 COMPOSPIMEPHALES MEASUREMENT

TRP6C 1 0 PERMIT 43.2 '** PERCENT SEMI COMPOS

EFFLUENT GROSS REQUIREMENT MIMINUM ANNUALSAMPLE

MEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

SAMPLEMEASUREMENT

PERMITREQUIREMENT

NAMEITITLE PRINCIPAL EXECUTIVE OFFICER I Certify under penalty of law that this document and all attachments were prepared under my I TELEPHONE DATEd- irection or supervision in accordance with a system designed to assure that qualified personnel

John T. Carlin properly gather and •evaluate the information submitted. Based on my inquiry of the person orpersons who manage the system, or those persons directly responsible for gathering the ice President 4 %information, the information submitted is, to the best of my knowledge and belief, true, accurate, 423 843-700i 1 i.Sequoyah Site Vice President and complete. lam aware that there e sign..n"t fcf Subrinting iai-, informajion, SIGNAJIE OF PRINCIPAL EXECUTIVE Iincluding the possibility of fine and imprisonment for knowing violations. OFFICER OR AUTHORIZED AGENT AREA NUMBER MO DAY

TYPED OR PRINTED CODE


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

EPA Form 3320-1 (REV 3199) Pmvious editions maybe used Page 1 of I

TENNESSEE VALLEY AUTHORITYTOXICITY TEST REPORT

INTRODUCTION / EXECUTIVE SUMMARYReport Date: September 17, 2012

1. Facility / Discharger: Sequoyah Nuclear Plant / TVA

2. County / State: Hamilton / Tennessee

3. NPDES Permit #: TN0026450

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

5. Design Flow (MGD): 1,579

6. Receiving Stream: Tennessee River (TRM 483.6)

7. 1Q10: 3,.491

8. Outfall Tested: 101

9. Dates Sampled: August 12- 17, 2012

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

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

12. Test Dates: August 14 - 21, 2012

13. Test Type: Short-term Chronic Definitive

14. Test Species: Fathead Minnows (Pimrehales promelas)Daphnids (Ceriodaphnia dubia)

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

Pimephalespromelas: UV treated Outfall 101: 10.8, 21.6, 43.2, 86.4, 100UV treated Intake: 100

16. Permit Limit Endpoint (%): Outfall 101: IC,5 = 43.2%

17. Test Results: Outfall 101: Pitnephales promelas: IC 25 > 100%Ceriodaphnia dubia: IC25 > 100%

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

Page 1 of 99

18. Facility Contact: Brad Love Phone #: (423) 843-6714

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

20. Lab Contact: Jim Sumner Phone #: (828) 350-9364

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

22. Notes: Outfall 101 samples collected August 12 - 17, 2012, showed no toxic effectsto fathead minnows or daphnids. The resulting IC25 values, for both species, were> 100 percent. Exposure of minnows and daphnids to intake samples resulted in nosignificant difference from the controls during this study period.

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

Page 2 of 99

METHODS SUMMARY

Samples:

1. Sampling Point: Outfall 101. Intake

2. Sample Type: Composite

3. Sample Information:

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

Sample Time (ET) Time (ET) Temp. TRC* Time (ET)ID Collected Received (°C) (mg/L) Last Used By

101 08-12-12 0645 to 08-13-12 1540 0 .8, 0 .7t <0.10 08-14-12 121308-13-12 0545 08-15-12 1115

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

08-13-12 0605 08-15-12 1115

101 08-14-12 0700 to 08-15-12 1553 1.9 1.9t <0.10 08-16-12 111408-14-12 0700to 08-16-12 1114

Intake 08-15-12 0620 08-15-12 1553 2.0 <0.10 08-16-12 111408-16-12 00to 08-18-12 1112

101 08-17-12 0600 08-17-12 1338 2 .2, 2 .2 t <0.10 08-19-12 111508-17-12 0600 08-20-12 1114

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

08-17-12 0615 08-20-12 1114

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

4. Sample Manipulation: Samples from Outfall 10 1 and intake were warmed to test temperature(25.0 ± 1.0°C) in a warm water bath.

Aliquots of Outfall 101 and Intake samples were UV-treated through a40-watt Smart® UV Sterilizer (manufactured by Emperor Aquatics,Inc.) for 2 minutes.

Page 3 of 99

Pimephales promelasTest Organisms:

Ceriodaphnia dubia

In-house Cultures1. Source: Aquatox, Inc.

2. Age: 20 - 20.22 hours old < 24-hours old

Test Method Summary:

1. Test Conditions: Static, Renewal Static, Renewal

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

Moderately Hard Synthetic3. Control / Dilution Water: Moderately Hard Synthetic

4. Number of Replicates: 4

5. Organisms per Replicate: 10

6. Test Initiation: (Date/Time)Outfall 101

UV Treated Outfall 101

7. Test Termination: (Date/Time)Outfall 101

UV Treated Outfall 101

8. Test Temperature: Outfall 101:

10

I

08-14-12 1200 ET08-14-12 1213 ET

08-21-12 1106 ET08-21-12 1122ET

Mean = 24.7°C(24.3 - 25.1 0C)

08-14-12 1148 ET

08-21-12 1053 ET

Mean = 24.9°C(24.6 - 25.30C)

Test Temperature: UV-Treated Outfall 10 1: Mean = 24.8°C(24.2 - 25.1 0C)

9. Physical / ChemicMeasurements:

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

Statistics were performed according to methods prescribed by EPAusing ToxCalc version 5.0 statistical software (Tidepool ScientificSoftware. McKinnevville. CA).

10. Statistics:

Page 4 of 99

TOXICITY TEST RESULTS (see Appendix C for Bench Sheets)

1. Results of a Pimephales promelas Chronic/ 7-day Toxicity Test.(Genus species) (Type / Duration)

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

Test Percent SurvivingSolutions (time interval used - days)

(% Effluent) 1 2 3 1 4 [6 7Control 100 100 100 100 100 100 10010.8% 100 100 100 100 100 100 10021.6% 100 100 100 100 100 100 10043.2% 100 100 100 100 100 100 10086.4% 100 100 100 100 100 100 100100.0% 100 100 100 100 100 100 100Intake 100 100 100 100 100 100 95

Mean Dry Weight (mg)Test Solutions (replicate number)(%1Effluent) 1 2 J 3 4 I Mean

Control 0.583 0.615 0.592 0.544 0.58410.8% 0.535 0.669 0.686 0.649 0.63521.6% 0.608 0.559 0.626 0.686 0.62043.2% 0.582 0.563 0.487 0.614 0.56286.4% 0.614 0.555 0.563 0.690 0.606100.0% 0.532 0.673 0.506 0.571 0.571Intake 0.537 0.569 0.551 0.687 0.586

IC 25 Value: > 100% Calculated TU Estimates: < 1.0 TUc*Permit Limit: 43.2%

Permit Limit: 2.3 TUc95% Confidence Limits:

Upper Limit: NALower Limit: NA

*TUa = 100/LC50 : TUc = 100/IC25

Page 5 of 99


2. Results of a Ceriodaphnia dubia Chronic/ 7-day Toxicity Test.(Genus species) (Type / Duration)

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

Percent SurvivingTest (time interval used - days)

Solutions(% Effluent) 6

Control 100 100 100 100 100 100 10010.8% 100 100 100 100 100 100 100

21.6% 100 100 100 100 100 100 100

43.2% 100 100 100 100 100 100 100

86.4% 100 100 100 100 100 100 100

100.0% 100 100 100 100 100 100 100

Test Solutions Reproduction (#young/female/7 days)Test Soluetis Data (replicate number)(% Effluent) 1 2 J3 ] 4 I 5 6 [ 7 1 8 [ 9 110 [Mean

Control 30 33 29 33 31 28 34 32 30 32 31.2

10.8% 36 34 33 33 35 35 34 34 31 32 33.7

21.6% 36 31 33 31 30 37 35 33 32 30 32.8

43.2% 37 34 34 31 31 31 37 34 34 35 33.8

86.4% 34 34 36 34 35 33 34 35 36 37 34.8

100.0% 38 37 37 34 32 38 35 39 37 38 36.5

IC 25 Value: > 100% Calculated TU Estimates: < 1.0 TUc*Permit Limit: 43.2%

Permit Limit: 2.3 TUc95% Confidence Limits:


*TUa = 1 00/LC5 0: TUc = 100/IC25

Page 6 of 99


2. Results of a Ceriodaphnia dubia Chronic/ 7-day Toxicity Test.(Genus species) (Type / Duration)

Conducted August 14 - 21, 2012 using water from Intake

Percent SurvivingTest (time interval used - days)

Solutions 1 1 5 611(% Effluent) 1 2 13 4 5

Control 100 100 100 100 100 100 100

Intake 100 100 100 100 100 100 100

Test Solutions Reproduction (#young/female/7 days)

(%Effluent) Data (replicate number)1 2 3 4 [ 5 [ 6 7 8 9 10 Mean

Control 30 34 34 31 32 29 32 31 29 31 31.3Intake 33 29 33 33 33 36 33 32

IC 25 Value: > 100% Calculated TU Estimates: < 1.0 TUc*Permit Limit: N/A

Permit Limit: N/A95% Confidence Limits:


*TUa = 1 00/LC50: TUc = 100/IC25

Page 7 of 99

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

3. Results of a Pimephales promelas Chronic/ 7-day Toxicity Test.(Genus species) (Type / Duration)

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

Test Percent SurvivingSolutions (time interval used - days)

(% Effluent) 1 ] 2 3 4 5 6 [ 7

Control 100 100 100 100 100 100 100

10.8% 100 100 100 100 100 100 100

21.6% 100 100 100 100 100 100 100

43.2% 100 100 100 100 100 100 100

86.4% 100 100 100 100 98 98 98

100.0% 100 100 100 100 100 100 100

Intake 100 100 100 100 100 100 100

Mean Dry Weight (mg)Test Solutions(rpiaenm )(% Effuent)replicate number)

(%Effluent) 1 [ 2 3 ] 4 Mean

Control 0.510 0.488 0.587 0.553 0.53510.8% 0.673 0.559 0.631 0.594 0.614

21.6% 0.599 0.524 0.547 0.582 0.56343.2% 0.592 0.516 0.548 0.588 0.56186.4% 0.509 0.527 0.551 0.448 0.509100.0% 0.550 0.565 0.526 0.494 0.534

Intake 0.62.1 0.573 0.647 0.482 0.581

IC 25 Value: > 100% Calculated TU Estimates: < 1.0 TUc*

95% Confidence Limits:Upper Limit: NALower Limit: NA

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

REFERENCE TOXICANT TEST RESULTS (see Appendix A and D)

Species Date Time 1 Duration I Toxicant I Results (IC25) IPimephales promelas August 14 - 21, 2012 1225

Ceriodaphnia dubia August 07 - 14, 2012 1045

KC1 0.81 g/L

NaCl 1.08 g/L

Page 8 of 99

(0(D(001%to

PHYSICAL/CHEMICAL SUMMARYWater Chemistry Mean Values and Ranges for Pitnephales promelas and Ceriodaphnia dubia Tests, Non-treated Sequoyah Nuclear Plant (SQN) Outfall 101

performed August 14-21, 2012.

Test Sample ID Temperature (C) Dissolved Oxygen (mgfL) OH (S.U.) Conductance Alkalinity Hardness Total Residual

Initial Final Initial Final Initial Final (mtmhos/cm) (mg/L CaCO3) (mg/L CaCO3 ) Chlorine (mglL)

Control 24.8 24.7 7.7 7.9 7.55 7.50 311 62 8924.7 - 24.9 24.6 - 25.1 7.6 - 7.9 7.8 - 8.0 7.43 - 7.67 7.36 - 7.59 302 - 323 62 - 62 88 - 92

10.8% 24.8 24.6 7.9 7.8 7.52 7.46 29124.7 - 25.0 24.3 - 25.0 7.7 - 8.0 7.6 - 8.0 7.44 - 7.62 7.33 - 7.55 285 - 298

21.6% 24.8 24.6 7.9 7.8 7.54 7.44 27824.7 - 25.0 24.4 - 24.9 7.7 - 8.0 7.6 - 8.0 7.44 - 7.62 7.33 - 7.55 272 - 283

4. 24.8 24.6 7.9 7.8 7.53 7.44 25243213 24.7 - 25.0 24.4 - 24.8 7.7 - 8.0 7.6 - 8.0 7.45 - 7.61 7.34 - 7.52 246 - 259

24.8 24.6 7.9 7.8 7.51 7.42 198S 86.4%

24.7 - 25.0 24.3 - 24.8 7.6 - 8.0 7.6 - 7.9 7.45 - 7.60 7.31 - 7.52 192 - 205

100.0% 24.9 24.6 7.9 7.9 7.51 7.46 182 71 72 < 0. 1024.8 - 25.0 24.4 - 24.8 7.7 - 8.1 7.7 - 8.1 7.42 - 7.60 7.32 - 7.57 177 - 186 70 - 72 71 - 74 <0.10- <0.10

Intake 24.9 24.6 8.0 8.0 7.51 7.45 182 69 73 < 0.1024.7 - 25.0 24.4 - 24.7 7.7 - 8.2 7.9 - 8.0 7.43 - 7.61 7.33 - 7.54 176 - 186 68 - 70 72 - 74 <0.10- <0.10

Control 24.8 25.1 7.7 8.0 7.55 7.51 311 62 8924.7 - 24.9 25.0 - 25.2 7.6 - 7.9 7.8 - 8.2 7.43 - 7.67 7.28 - 7.59 302 - 323 62 - 62 88 - 92

10.8% 24.8 25.0 7.9 8.0 7.52 7.51 29124.6 - 25.0 24.8 - 25.2 7.7 - 8.0 7.8 - 8.2 7.44 - 7.62 7.29 - 7.59 285 - 298

2!1 24.8 25.0 7.9 8.0 7.54 7.52 27824.7 - 25.0 24.8 - 25.1 7.7 - 8.0 7.8 - 8.2 7.44 - 7.62 7.30 - 7.60 272 - 283

4.% 24.9 24.9 7.9 8.0 7.53 7.52 252x 43.2%24.7 - 25.0 24.8 - 25.3 7.7 - 8.0 7.9 - 8.2 7.45 - 7.61 7.30 - 7.61 246 - 259

8 24.9 25.0 7.9 8.0 7.51 7.51 198• 86.4%___ 24.7 - 25.1 24.8 - 25.2 7.6 - 8.0 7.9 - 8.2 7.45 - 7.60 7.28 - 7.61 192 - 205

100.0% 25.0 24.9 7.9 8.0 7.51 7.52 182 71 72 < 0.1024.8 - 25.1 24.8 - 25.2 7.7 - 8.1 7.9 - 8.3 7.42 - 7.60 7.29 - 7.63 177 - 186 70 - 72 71 - 74 <0.10- <0.10

Intake 25.0 24.9 8.0 8.1 7.51 7.52 182 69 73 < 0.1024.8 - 25.1 24.7 - 25.1 7.7 - 8.2 8.0 - 8.2 7.43 - 7.61 7.30 - 7.64 176 - 186 68 - 70 72 - 74 <0.10- <0.10

Overall temperature (C)Pimephales promelasCeriodaphnia dubia

Average24.724.9

Minimum24.324.6

Maximum25.125.3

0,,

O-I'(0

PHYSICAIJCHEM ICAL SUM MARYWater Chemistry Mean Values and Ranges for Pitnephalespromelas Test, UV-treated Sequoyah Nuclear Plant (SQN) Outfall 101

performed August 14-21, 2012.

Test Sample ID Temperature (C) Dissolved Oxygen (mg/L) pH (S.U.) Conductance Alkalinity Hardness *Total Residual

Initial Final Initial Final Initial Final (pmhos/cm) (mg/L CaCO3) (mg/L CaCO3 ) Chlorine (ng/L)

Control 24.8 24.7 7.9 7.8 7.59 7.49 304 61 8424.8 - 24.9 24.5 - 24.9 7.8 - 8.1 7.6 - 7.9 7.50 - 7.65 7.34 - 7.61 298 - 316 61 - 62 84 - 84

10.8% 24.9 24.7 7.9 7.8 7.60 7.48 29124.8 - 25.0 24.5 - 25.0 7.9 - 8.0 7.6 - 7.8 7.50 - 7.66 7.34 - 7.57 282 - 298

24.9 24.7 7.9 7.8 7.59 7.48 27921.6%21.6% 24.8 - 25.1 24.6 - 24.8 7.9 - 8.0 7.7 - 7.9 7.50 - 7.66 7.34 - 7.58 271 - 284C, 24.9 24.7 7.9 7.8 7.58 7.47 25443.2%

&1____ 24.8 - 25.1 24.6 - 24.8 7.9 - 8.0 7.7 - 7.9 7.49 - 7.66 7.33 - 7.58 246 - 258

86.4% 25.0 24.7 8.0 7.8 7.56 7.46 20324.9 - 25.1 24.3 - 24.8 7.9 - 8.1 7.5 - 7.9 7.48 - 7.63 7.33 - 7.57 197 - 207

100.0% 25.1 24.6 8.0 7.8 7.55 7.45 184 70 72 <0.1025.0 - 25.1 24.2 - 24.7 7.8 - 8.3 7.5 - 7.9 7.47 - 7.62 7.31 - 7.55 178 - 188 70 - 70 72 - 72 <0.10 - <0.10

Intake 25.0 24.6 8.1 7.9 7.54 7.46 184 70 72 < 0. 1024.8 - 25.1 24.5 - 24.9 7.9 - 8.2 7.7 - 8.0 7.46 - 7.61 7.32 - 7.58 178 - 191 70 - 70 71 - 72 <0.10 - <0.10

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

Overall temperature (C) Average Minimum MaximumPimephales promelas 24.8 24.2 25.1

SUMMARY /CONCLUSIONS

Outfall 101 samples collected August 12 - 17, 2012, showed no toxic effects tofathead minnows or daphnids. The resulting IC25 values, for both species, were> 100 percent. Exposure of minnows and daphnids to intake samples resulted in nosignificant difference from the controls during this study period.

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

Page 11 of 99

Appendix A

ADDITIONAL TOXICITY TEST INFORMATION

SUMMARY OF METHODS

1. Pimephales promelas

Tests were conducted according to EPA-82 I -R-02-013 (October 2002) using fourreplicates, each containing ten test organisms, per treatment. Test vessels consisted of 500-mL plastic disposable cups, each containing 250-mL of test solution.

2. Ceriodaphnia dubia

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

DEVIATIONS / MODIFICATIONS TO TEST PROTOCOL

1. Pinephales promelas

None


None

DEVIATIONS / MODIFICATIONS TO PRETEST CULTURE OR HOLDING OF TESTORGANISMS

1. Pimephales promelas

None


None

Page 12 of 99

PHYSICAL AND CHEMICAL METHODS

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

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

3. Temperature was measured by SM 2550 B.

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

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

6. Conductance was measured by SM 2510 B.

7. Alkalinity was measured by SM 2320 B.

8. Total hardness was measured by SM 2340 C.

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

QUALITY ASSURANCE

Toxicity Test Methods: All phases of the study including, but not limited to, samplecollection, handling and storage,. glassware preparation, test organismculturing/acquisition and acclimation, test organism handling during test, andmaintaining appropriate test conditions were conducted according to the protocol asdescribed in this report and EPA-821-R-02-013. Any known deviations were notedduring the study and are reported herein.

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

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

2. Standard Toxicant: Potassium Chloride (KCI crystalline) for Pimnephalespromelas.Sodium Chloride (NaCl crystalline) for Ceriodaphnia dubia.

3. Dilution Water Used: Moderately hard synthetic water.

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

Page 13 of 99

REFERENCES

1. NPDES Permit No. TN0026450.

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

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

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

Page 14 of 99

Sequoyah Nuclear Plant BiomonitoringAugust 14 - 21, 2012

Appendix B

Diffuser Discharge Concentrations of Total Residual Chlorine,

Diffuser Discharge Concentrations of Chemicals Used toControl Microbiologically Induced Corrosion and Mollusks

During Toxicity Test Sampling

Page 15 of 99

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) DischargeConcentrations of Chemicals Used to Control Microbiologically Induced Corrosion

Mollusks, During Toxicity Test Sampling,March 12, 1998 - August 17, 2012

Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat- H-130MHypochlorite mg/L mg/L mg/L mg/L PF mg/L

mgIL TRC Phosphate Copolymer DMAD mg/L QuatTRC Azole

03/12/1998 0.01603/13/1998 0.01503/14/1998 0.01303/15/1998 0.03003/16/1998 0.01303/17/1998 0.02003/18/1998 0.018 - - - - - -

09/08/1998 0.015 - 0.014 0.005 - - 0.02109/09/1998 0.003 - 0.031 0.011 - - -

09/10/1998 0.014 - 0.060 0.021 - -09/11/1998 0.013 - 0.055 0.019 - -

09/12/1998 < 0.001 - 0.044 0.01509/13/1998 < 0.001 - 0.044 0.01509/14/1998 0.008 0.044 0.015 102/22/1999 < 0.001 - -

02/23/1999 0.00502/24/1999 0.00902/25/1999 0.01202/26/1999 0.00802/27/1999 < 0.00102/28/1999 < 0.001 - - I I08/18/1999 - 0.015 0.069 0.024 0.00608/19/1999 - 0.012 0.068 0.024 - -

08/20/1999 - 0.023 0.070 0.024 - 0.12008/21/1999 - 0.022 0.068 0.024 - -08/22/1999 - 0.022 0.068 0.024 - -

08/23/1999 - 0.025 0.068 0.024 0.006 -

08/24/1999 - 1 0.016 0.067 0.023 0.020 -

Page 16 of 99

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations ofChemicals Used to Control Microbiologically Induced Corrosion Mollusks, During

Toxicity Test Sampling,March 12, 1998 - August 17, 2012

Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat- H-130MHypochlorite mg/L mgiL mg/L mg/L PF mg/L

mg/L TRC Phosphate Copolymer DMAD mg/L. QuatTRC Azole

01/31/2000 -< 0.002 0.026 0.00902/01/2000 - 0.011 0.026 0.028 -

02/02/2000 - 0.028 0.026 0.009 0.00602/03/2000 - 0.008 0.027 0.009 - -

02/04/2000 - 0.006 0.027 0.009 0.005 0.10902/05/2000 -< 0.002 0.027 0.009 - -02/06/2000 - < 0.002 0.027 0.009 - I07/26/2000 -< 0.0057 0.055 0.019 - -

07/27/2000 - 0.019 0.055 0.019 -07/28/2000 - 0.0088 0.053 0.018 0.004 0.10807/29/2000 -< 0.0088 0.055 0.019 - -07/30/2000 -< 0.0076 0.055 0.019 -07/31/2000 -< 0.0152 0.055 0.019 0.00608/01/2000 - < 0.0141 0.055 0.019 0.005 112/11/2000 - 0.0143 0.025 0.020 0.00512/12/2000 - 0.0092 0.025 0.020 0.00512/13/2000 -< 0.0120 0.025 0.020 - -

12/14/2000 -< 0.0087 0.025 0.020 - -

12/15/2000 - 0.0120 0.025 0.020 0.005 -

12/16/2000 - < 0.0036 0.025 0.020 - -

12/17/2000 -< 0.0036 0.025 0.020 - -

08/26/2001 - 0.017 0.06 0.021 0.006 -

08/27/2001 - <0.0096 0.06 0.021 0.005 - 0.02108/28/2001 - <0.0085 0.06 0.021 - --

08/29/2001 - <0.0094 0.059 0.020 0.005 - 0.02108/30/2001 - <0.0123 0.06 0.021 0.005 -08/31/2001 - <0.005 0.059 0.020 - -

11/25/2001 - <0.0044 -- -

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

11/27/2001 - 0.0137 0.023 0.019 0.007 -

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

11/29/2001 - 0.0132 0.024 0.02 0.007 -

11/30/2001 - < 0.0043 0.024 0.02 -- I12/09/2001 - <0.0042 - -12/10/2001 - <0.004212/11/2001 - <0.0104 - - -12/12/2001 - 0.0128 0.024 0.02 0.008 -

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

12/14/2001 - 0.0134 0.024 0.02 0.007 - 1

Page 17 of 99




mg/L TRC Phosphate Copolymer DMAD mg/L QuatTRC Azole

01/02/2002 -< 0.0079 0.023 0.02 0.00601/03/2002 -< 0.0042 0.023 0.014 -

01/04/2002 - 0.0124 0.024 0.014 0.00901/05/2002 -< 0.0042 - - -

01/06/2002 - < 0.0042 - - -

01/07/20021 - <'0.0089 0.024 0.014 0.006 102/24/2002 - < 0.004 - - -

02/25/2002 -< 0.004 0.023 0.023 -

02/26/2002 - 0.0143 0.023 0.023 0.00702/27/2002 -< 0.0041 0.023 0.023 -

02/28/2002 -< 0.0041 0.024 0.00803/01/2002 - < 0.0041 0.024 0.008 105/05/2002 - - - -

05/06/2002 - - 0.058 0.02 0.01405/07/2002 - - 0.058 0.02 0.01505/08/2002 - - 0.056 0.019 -

05/09/2002 - - 0.057 0.02 0.01405/10/2002 - 0.056 0.019 -- I08/04/2002 - <0.0058 - - --

08/05/2002 - <0.0058 0.053 0.018 - 0.02508/06/2002 - 0.0092 0.053 0.018 - -

08/07/2002 - <0.0107 0.055 0.019 0.00708/08/2002 - <0.0061 0.055 0.019 -

08/09/2002 - 0.0152 0.054 0.018 0.008 110/06/2002 - <0.00497 - - -

10/07/2002 - 0.0153 0.054 0.018 0.00910/08/2002 - <0.0092 0.054 0.018 0.00710/09/2002 - 0.0124 0.053 0.018 0.00910/10/2002 - 0.0134 0.054 0.018 0.00910/11/2002 - <0.0042 0.054 0.018 -

01/12/2003 - <0.0035 - - -

01/13/2003 - <0.006 0.025 0.019 0.00901/14/2003 - <0.01 18 0.026 0.02001/15/2003 - <0.0063 0.026 0.020 0.00901/16/2003 - <0.0034 0.026 0.02001/17/20031 - <0.0034 0.026 0.009 -

04/06/2003 - <0.0073 - - -

04/07/2003 - <0.0189 - 0.021 -

04/08/2003 - <0.0117 - 0.021 -

04/09/2003 - <0.0139 - 0.021 0.01604/10/2003 - <0.0113 - 0.021 0.01804/11/2003 - <0.0073 - 0.022 - - -

Page 18 of 99

Table B-1 (continued). Sequoyah Nuclear Plant Diffuser (Outfall 101) DischargeConcentrations of Chemicals Used to Control Growth of Microbiologically Induced

Bacteria and Mollusks, During Toxicity Test Sampling,March 12, 1998 - August 17, 2012


mg/L TRC Phosphate Copolymer DMAD mg/L QuatTRC Azole

06/15/2003 -< 0.0045 - - - -06/16/2003 -< 0.0037 0.057 0.020 - - 0.02206/17/2003 - < 0.0048 0.041 0.014 - - 0.02406/18/2003 - < 0.0048 0.041 0.014 - - 0.02406/19/2003 - < 0.0085 0.058 0.020 - - 0.02506/20/2003 - < 0.0048 0.058 0.020 1 - 0.02508/03/2003 - <0.0050 - - -

08/04/2003 - <0.0050 0.058 0.020 -08/05/2003 - <0.0051 0.057 0.020 - 0.02508/06/2003 - <0.0084 0.057 0.020 - 0.02508/07/2003 - 0.0129 0.057 0.020 - 0.02408/08/2003 - 0.0153 0.057 0.020 0.009 - -

10/05/2003 - <0.0043 0.057 0.020 - - -10/06/2003 - <0.0043 0.057 0.020 - - 0.02510/07/2003 - <0.0090 0.057 0.020 - - 0.02510/08/2003 - <0.0106 0.057 0.020 - - 0.02510/09/2003 - 0.0181 0.026 0.022 - - 0.02510/10/2003 - 0.0183 0.026 0.024 0.009 -

02/01/2004 - 0.0093 0.027 0.009 -02/02/2004 - <0.0034 0.026 0.00902/03/2004 - <0.0034 0.026 0.009 -02/04/2004 - 0.0124 0.026 0.009 0.00902/05/2004 - <0.0034 0.026 0.009 - -02/06/2004 - 0.0105 0.026 0.009 0.010 - -

05/04/2004 - <0.0123 0.026 0.019 - - 0.02505/05/2004 - <0.0144 0.026 0.014 0.009 - 0.02505/06/2004 - <0.0146 0.037 0.013 - - 0.02505/07/2004 - 0.0227 0.058 0.020 0.009 - 0.02505/08/2004 - 0.016 0.060 0.021 - - -05/09/2004 - <0.0104 0.058 0.020

07/04/2004 - 0.0217 0.057 0.019 -

07/05/2004 - <0.0085 0.057 0.020 0.00907/06/2004 - <0.0077 0.058 0.020 - - 0.03107/07/2004 - 0.0252 0.056 0.019 - - 0.03107/08/2004 0.0223 0.057 0.019 0.009 - -

07/09/2004 0.0182 0.057 0.020 0.009 -

Page 19 of 99



Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-PF H-130M Nalco t-150MHypochlorit mg/L mg/L mg/L mg/L mg/L mg/L 73551 mg/L

e TRC Phosphate Copolymer DMAD Azole Quat mg/L Quatmg/L EO/POTRC

11/07/2004 - <0.0187 0.000 0.014 - -

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

11109/2004 - <0.0233 0.048 0.016 - - 0.04111/10/2004 - <0.0149 0.047 0.016 - - 0.04111/11/2004 - <0.0149 0.049 0.017 -1 - 0 .04 3

111/12/2004 - 1 <0.0253 0.048 1 0.017 0.04202/06/2005 <0.0042 0.028 0.010 -02/07/2005 - <0.0116 0.028 0.010 - 0.00702/08/2005 - <0.0080 0.028 0.010 I02/09/2005 - 0.0199 0.028 0.01002/10/2005 - <0.0042 0.028 0.01002/11/2005 - 0.0155 0.028 0.010 0.00706/05/2005 -0.0063 -06/06/2005 - 0.0043 - - 0.03706/07/2005 - 0.0103 - - 0.03706/08/2005 - 0.0295 - - 0.03706/09/2005 - 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 0.03607/19/2005 - 0.0163 0.026 0.009 - 0.03607/20/2005 - 0.0209 0.026 0.009 - 0.014 0.03607/21/2005 - 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 - - 0.03511/02/2005 - 0.0104 - - 0.03611/03/2005 - 0.0117 - - 0.03611/04/2005 - 0.0165 - - 0.035

11/14/2005 - 0.0274 - - -

11/15/2005 - 0.0256 -

11/16/2005 - 0.0234 --

11/17/2005 - 0.0231 -

11/18/2005 - 0.0200 --

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

Page 20 of 99

Table B-1 (continued). Sequoyah Nuclear Plant Diffuser (Outfall 101) DischargeConcentrations of Chemicals Used to Control Growth of Microbiologically Induced

Bacteria and Mollusks, During Toxicity Test Sampling,March 12, 1998 - August 17, 2012

Date Sodium Towerbrom PCL-222 PCL-401 CL-363 Cuprostat-PF H-130M Nalco H-150M MSWHypochlorite mg/L mg/L mg/L mg/L mg/L mg/L 73551 mg/L 101

mgiL TRC Phosphate Copolymer DMAD Azole Quat mg/L Quat mg/L

TRC EO/PO Phosphate11/12/2006 - 0.0055 - - -

.11/13/2006 - 0.0068 -- 0.03711/14/2006 - 0.0143 -- 0.03711/15/2006 - 0.0068 -- 0.03711/16/2006 - 0.0267 -- 0.03711/17/2006 - 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 - - - - 0.01505/29/07 - - - - 0.036 0.01505/30/07 - 0.0084 - - 0.017 0.036 0.01505/31/07 - 0.0103 - - - 0.036 0.01506/01/07 - 0.0164 - - 0.017 0.036 0.01506/02/07 - 0.0305 - - - - 0.015

12/02/07 - 0.0241 - - -

12/03/07 - 0.0128 - -

12/04/07 - 0.0238 - -

12/05/07 - 0.0158 - -

12/06/07 - 0.0162 - -

12/07/07 - 0.0175 --

04/13/08 - 0.0039 -

04/14/08 - 0.0124 -

04/15/08 - 0.0229 -04/16/08 - 0.0143 -04/17/08 - 0.0120 -04/18/08 - 0.0149 -

10/26/08 - 0.0260 -10/27/08 - 0.0151 - 0.017 -

10/28/08 - 0.0172 - - 0.041 -10/29/08 - 0.0154 - 0.018 0.041 0.03010/30/08 - - - - 0.041 0.03010/31/08 - 0.0086 -- 0.041 0.030

Page 21 of 99

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations ofChemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test

Sampling,March 12, 1998 - August 17, 2012

Date Sodium Towerbrom PCL- PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSWHypochlorite mg/L 222 mg/L mg/L A-PF mg/L mg/L 73551 CT1300 mg/L 101

mg/L TRC mg/L Copolymer DMAD Azole Quat mg/L mg/L Quat mg/LTRC Phosph EO/PO Quat Phosphate

ate02/08/09 - 0.0197 - - - - 0.017 - -02/09/09 - 0.0237 - - - - 0.017 - -02/10/09 - 0.0104 - - - 0.021 - -02/11/09 - 0.0155 - - - 0.017 - -02/12/09 - 0.0106 - - - 0.017 -

02/13/09 - - -

05/10/09 - 0.0129 - - -05/11/09 - 0.0415 -- 0.0446 -

05/12/09 - 0.0053 - - 0.0396 -

05/13/09 - 0.0049 - - 0.0396 -

05/14/09 - <0.0141 - - 0.0397 -

05/15/09 - <0.0160 - - -

11/15/09 - 0.025 -- - -

11/16/09 - 0.0152 -- - -

11/17/09 - 0.0255 -- - -

11/18/09 - 0.0306 - -.11/19/09 - 0.0204 - -- -

11/20/09 - 0.0093 - --

05/09/10 - 0.0192 - --

05/10/10 - 0.0055 - --

05/11/10 - 0.0100 - - - 0.039 -05/12/10 - 0.0171 - - - 0.039 -05/13/10 - 0.0041 - - - 0.039 -05/14/10 - 0.0099 - - - 0.039 -

Page 22 of 99

Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations ofChemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test

Sampling,March 12, 1998 - August 17, 2012

Date Sodium Towerbrom PCL- PCL-401 CL-363 Cuprostat H-130M Nalco Spectrus H-150M MSW FloguardHypo- mg/L 222 mg/L mg/L -PF mg/L mg/L 73551 CT1300 mg/L 101 MS6236chlorite TRC mg/L Copoly- DMAD Azole Quat mgiL mg/L Quat mg/P , mgPmg/L Phos- meC EO/PO Quat Phos.- PhosphateTRC phate phate

10/31/1011/01/10 0.012211/02/10 0.011211/03/10 0.016311/04/10 0.010711/05/10 0.0132

05/01/2011 - -

05/02/2011 - 0.0405/03/2011 -- 0.0405/04/2011 - 0.0155 0- 0.0405/05/2011 - 0.0179 - 0.0405/06/2011 - 0.0089 - - -

11/06/2011 - 0.016811/07/20.11 - 0.022511/08/2011 - 0.014111/09/2011 - 0.023911/10/2011 - 0.024211/11/2011 - 0.0231

05/06/2012 - -

05/07/2012 -

05/08/2012 - - 0.04105/09/2012 - 0.0145 - 0.04105/10/2012 - 0.0298 -- 0.04105/11/2012 - 0.0174 -

08/12/2012 - - - - 0.02908/13/2012 - 0.0256 - 0.028 0.037 - 0.02908/14/2012 - 0.0209 - - 0.037 - 0.02908/15/2012 - 0.0279 - 0.028 - - 0.02908/16/2012 - 0.0076 - - 0.02908/17/2012 - 0.0446 - 0.032

Page 23 of 99


Appendix C

Chain of Custody Records andToxicity Test Bench Sheets

Page 24 of 99

BIOMONITORING CHAIN OF CUSTODY RECORD Page _1_ of-1-'I U

Rent: TVA

lRkject Name: Sequoyah NP Toxicity

I%. Number: N/A

Facility Sampled: Sequoyah NP

NPDES Number: TN0026450

Collected By: pw-k% 1 ej r -

Environmental Testing Solution, Inc.

351 Depot Street.

Asheville, NC

28801

Phone: 828-350-9364

Delivered By (Circle One):

FedEx UPS Bus Client

Other (specify):

General Comments:

roolý 5•fYo '-a -f ,P/1 .4+. 0 CCo, o,

.Fax: 828-350-9368

Container .' .0.Field Identification Grab/Comp. Collection Date/Time& Flow Rain Event?Sample Description V. CVolume (MGD) as Appropriate Laboratory Use

'__ ___ Collected(,_ R ... .~-. s n hT . E Tg.N r . . iP". " . . .... . ." " p rIf Y es A rrival T emp. B y T im e A ppear-

_______tt~~MW __ Inches Number ance8 " 12 "t1- 3 J- 2I qi••. -:: Ii• . ':•.• -,: :::::;:•: ,:•,:, ': . ...

SQN-101-TOX Comp " Lj 2(2.5gal) 1'I/ A o2/1,oi _•-._ 17'_. .

SQN-INT-TOX Comp 1 •.x- Ot63 ' 1(2.5gal) )2?ol;: -.! i i . 0Q

Sample Custody - Fill In From Top Down . " wCturr se' iwpr-t,•'c. SAmAkes. ae.cewebl,

Relinquished By (Signature): Date/Time Received By (Signature): Date/Time , WLC.Pt & . A

• 0I-I•o t'v -/// .o4:.0S 0 E

Instructions: Clients should fill in all areas except those in the "Laboratory Use" block. Biomonitoring samples are preserved by storing them at-6°C and shipping them in ice. The hold time for each sampleis 36 hours 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 shippedovernight on Friday via FedEx or UPS must be marked for Saturday delivery or they will not arrive until the following Monday.

MIS1.11 ýIfl w

Whole Efflue' -)ample Receipt Log Page .Z

*Sample temperature performed usingSample Receiving Thermometer. SN 6338

0

(0

Date Time Received Received *Sa m ple' Project Sample

Received Received by from Temp. ( 0C) number number Sample name and description State Comments

08-13-12 1540 J. Sumner Sonic Delivery 0.8,0.7 8207 120813 .01 TVA/ SON Outfall 101 TN08-13-12 1540 J. Sumner Sonic Deliver 0.9 8207 120813 .02 TVA / SQN Intake TN

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

BIOMONITORING CHAIN OF CUSTODY RECORD Page 1 of 1.

2ent: TVA Environmental Testing Solution, Inc. Delivered By (Circle One):

Wlject Name? Sequoyah NP Toxicity 351 Depot Street. FedEx UPS Bus Client

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

2880 General Comments:Facility Sampled: Sequoyah NP 28801 Teneral Comments:

NPDES Number: TN0026450 Phone: 828-350-9364 ,, ,,ke a 00 ,Jo,

Collected By: t, 0 fl"Ž•. Fax: 828-350-9368

Container " .. A .. ,.Field Identification Number & Flow RainEent?FildIdntfiaton/ Grab/Comp. Collection Date/Time Rain Event?

Sample Description Volume (MGD) .""aLabspratoo aU)seCollected (Mark as Appropriate)

.. .,. . Start!~~ EndYes If Yes, No Trace ETS Log Arrival Tempp. By, Time• Appear-•i•W ° • • ' • ..t ? ?YrJ.Str E Inches _.._. Number, :'_""" ance

SQN-101-TOX Comp 0700A 6" 0. I" 2(2.5gal) Y/* l [• 0 " - ! .

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

,__ __ ___ __Sample Custody - Fill In From Top Down 4 uokmR , s -s J r5 RAE1 i.•.Relinquished By (Signature): Date/Time Received By (Signature): Date/Time AtU ýt".eS.

-T A ,hr-1- /03-e Skl._c C'r6oC

Instructions: Clients should fill in all areas except those in the "Laboratory Use" block. Biomonitoring samples are preserved by storing them at 6°C and shipping them in ice. The hold time for each sampleis 36 hours 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 shippedovernight on Friday via FedEx or UPS must be marked for Saturday delivery or they will not arrive until the following Monday.

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

~Iiii(0

0

(0(0

*Sample temperature performed using Sample Receiving Thermometer SN 6338

Date Time Received Received *Sample Project SampleReceived Received by from Temp. (0C) number number Sample name and description State Comments

08-15-12 0945 K. Keenan Fed - Ex 1.3 8215 120815 .01 CORR ERI - AAF McQuay International NC08-15-12 0945 K. Keenan Fed - Ex 0.7 8216 120815 .02 Statesville Analytical - Cleveland WWTP NC08-15-12 0945 K. Keenan Fed - Ex 5.5 8217 120815.03 United Water - Enfield WWTP NC08-15-12 0945 K. Keenan Fed - Ex 5.5 8218 120815.04 United Water- Scotland Neck WWTP NC08-15-12 0945 . Keenan Fed - Ex 0.3 8219 120815 .05 Craven County Wood Energy NC08-15-12 0945 K. Keenan Fed - Ex 0.7 8220 120815 .06 Duke Energy - McGuire NS - Outfall 002 NC08-15-12 0945 K. Keenan Fed - Ex 0.6 8221 120815 .07 Progress Energy - Shearon Harris E & E NC08-15-12 0945 K. Keenan Fed - Ex 0.5 8222 120815 .08 Carolina Beach WWTP NC08-15-12 0945 K. Keenan UPS 2.0 8223 120815 .09 Morehead City WWTP NC08-15-12 1553 J. Sumner TVA Courier 1.9/1.9 8207 120815 .10 WA - SQN - 101 TN08-15-12 1553 J. Sumner TVA Courier 2.0 8207 120815 .11 ]TVA - SQN - Intake ITN I


BIOMONITORING CHAIN OF CUSTODY RECORD Page _1- of -1-

4lent: TVA

Wject Name: Sequoyah NP Toxicity

Q•. Number: N/AccFacility Sampled: Sequoyah NP

NPDES Number: TN0026450

Environmental Testing Solution, Inc.

351 Depot Street.

Asheville, NC

28801

Phone: 828-350-9364

FedEx UPS Bus Client

Delivered By (Circle One):

Other (specify):__________

General Comments:

Q t- 0,( &,a/ 7-V

Collected By: Iw.;'f r6 n3in.p Fax: 828-350-9368

Container '.- 1 8.',O.N um ber & Flow Rain Event?..:.... . ..: •..-::.•.... .. ... :..:: . ..... . ..Field Identification / Grab/Comp. Collection Date/Time VumeRain Event?.M.D.Sample Description Volume (MGD) (Mark as Appropriate) LaboratoryUsc

Collected•(Mark.as..ppr-p..iate. ::Laborator .Use

tW-:- " Sta...End ,t" .. • •I Stets Yes, No T ETS Log Arrival Temp. By Time, Appearý-______________ . _____ Start__ End_ _ _ ___._._ Yes Inches No Trace "Number .- ". ance .

SQN-101-TOX Comp 0 70o 67 Oo vr 2(2.5gal) / 1

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

Relinquished By (Signature): Date/Time Received By (Signature): Date/Time .SAM-s4 •

46t S-Pv. Date/Tim 15e,- ..--..- qr

Instructions: Clients should fill in all areas except those in the "Laboratory Use" block- Biomonitoring samples are preserved by storing them at 6°C and shipping them in ice. The hold time for each sampleis 36 hours 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 shippedovernight on Friday via FedEx or UPS must be marked for Saturday delivery or they will not arrive until the following Monday.

~0

0

CC(0

X ýrs-11 --Whole Efflue( '.ýample Receipt Log - Page 12S

*Sample temperature performed uslngSample Receiving Thermometer. SN 6338

Date Time Received Received *Sample Project Sample

Received Received by from Temp. C0C) number number08-17-12 0950 K. Keenan Fed - Ex 0.6 8215 120817 .01 CORR ERI - AAF McQuay International NC08-17-12 0950 K. Keenan Fed - Ex 0.6 8216 120817.02 Statesville Analytical - Cleveland WWTP NC08-17-12 0950 K. Keenan Fed - Ex 2.0 8217 120817.03 United Water - Enfield WWTP NC08-17-12 0950 K. Keenan Fed - Ex 2.0 8218 120817 .04 United Water - Scotland Neck WWTP NC08-17-12 0950 K. Keenan Fed - Ex 0.7 8219 120817 .05 Craven County Wood Energy NC08-17-12 1130 J. Sumner Murphy Courier 0.8 8226 120817 .06 Duke Energy - McGuire NS - Upper Composite NC08-17-12 1130 J. Sumner Murphy Courier 0.8 8226 120817 .07 Duke Energy - McGuire NS - Lower Composite NC08-17-12 1338 J. Sumner TVA Courier 2.2/2.2 8207 120817 .08 TVA - SQN - Outfall 101 TN08-17-12 1338 J. Sumner TVA Courier 2.4 8207 120817 .09 TVA - SON - Intake TN


:TSPage Iof 6

Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1000.0)Species: Pimephales promelas

Client: Tennessee Valley AuthorityFacility: Sequoyah Nuclear PlantNPDES #: TN0020168Project #: A iJ1

County: RheaOutfall: 101

Dilution preparation information: Comments:Dilution prep (%) 10.8 21.6 43.2 86.4 100Effluent volume (mL) 270 540 1080 2160 2500

Diluent volume (mL) 2230 1960 1420 340 0

TOW volume (mL) 2500 2500 2500 2500 2500

Test organism information: Test information:Organism age: 21) &ts O.z Randomizing template: JeLLOQ.Date and times organisms Oa • rh " Incubator number andwere born between: shelf location: __.,

Organism source: AYb)( O k P e- - Artemia CHM number: LIA 6S2Drying information for weightdetermination:

Transfer bowl information: pH = -1. to 0 S.U. Date / Time in oven: 06o.. 1I%Temperature = "S.•, 0C Initial oven temperature: lb, C

Average transfer volume: Date / Time out of oven: ______

0. MLOck Final oven temperature: J 'Total-drying time: ".t*4

Daily feeding and renewal information:

Day Date Morning feeding Afternoon feeding Test initiation, Sample numbers used MHSWrenewal, or batchtermination used

Time Analyst Time Analyst Time Analyst Outfall 101 Intake

2 -. . -•-L I% O. 14-r

0w5 VLOI 11.01 1701.01l.2 O&F1615 W- 1R. 11- M IS Oh s A, I % __1•__ _' • I . 0.16- m2 0|- ____'_. ______ _____$_5 " __ _\___ •l t•01l • '. 0 5. o-i'l1

6 0g**121. '- g,-Ift 11.09 I 7.0,f n. do, a6--i7'IL- - u-.4 -Ito(,

Control information: Acceptance -riteria Summary of test endpoints:% Mortality: 07, < 20% 7-day LCsO > I W7.Average weight per initial larvae: n3.SU W NOEC 1CCW7.Average weight per surviving larvae: 0. Sq > 0.25mg/larvae . LOEC > 1007.

ChV >1/MO.IC 25 I >1___T...

(I age 31 of 99

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

of 6

Species: Pimephales promelasClient: TVA / Sequoyah Nuclear Plant, Outfall 101, Non-treated Date: 0&. 1q.- I,."

Survival and Growth DataDay CONTROL 10.8% 21.6%

/A B C D E F G H I J K L

0 10DI(C/ 0 / /bo 10 o'o lio o/0o'ro0__01_ _ t(0 /0 tol/0 /0 .10 / /0 to(0 0

2 /0 to 10 lQ10 Ito (0 to t 10 to 10 (oSIQ0 10 10 10 to 10 10 10 10 10 'o /0

4_______ 10 1o NO0 1 to Ito( 10 10 ) t0 o 10 to2 /0 /0 10 10 /0 /0 /0 /0 /0 '0 /'0 (0

6 /0 /0 t0 10 (0 10 I0 10 (0 /0 /6 ID

6 11) to Io 'i o 101 10o fo o t0 to 'ro 107 1o t) /0 t 0 1O /lb C) 1 10 CO /

A = Pan weight (mg)Tray color code:: L.;!,%•ý ;t

Analyst: wIl.y AA 13.-s 14.z4 13.Z4 14.10 13.B, I3.6 13-38 14.zs lZ.'e IS. 14.14Date: OA.n-.t-z

B = Pan + Larvae weight (mg)Analyst: 3"Lt Lo•o %J.73 7.o.16 1I6V tq.q5 Zo.SS •$ Z.Sq 1q.17 z2*33 19Sq 11,93 "u.S-Date: 09-zl.17.

C = Larvae weight (mng) = B - A

Hand calculated. S M 14-1. SAI 6 6A0 . .09 svi (".m. 6..&6Analyst: _ k _III

Weight per initial number of larvae (mg)= C / Initial number of larvae

Hand calculated.Analyst: __

eV 0% No 'I i'~p 4.Average weight per Percent reductioninitial number of from control (%) 0.614larvae (mg)

- . - p - i - * -9- . -- i -

0 . ts -&.&7. 0.ADULD - •.•a7.

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

Comments:

ge 32 of 99

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

III

III*J.

I

V

IpI

w

I

JETSPage 3 of 6

Species: Pimephales promelasClient: TVA / Sequovah Nuclear Plant. Outfall 101. Non-treated Date: 9O&. Iq- I*.--

Survival and Growth DataDay 43.2% 86.4% 100%

M N 0 P Q R S T U V W X0 0 to I olo to l to to to IO U~ ( ~ 1,C) to1 O 10( to to 10 10 / /0 /(0 I0 ( 0 I)10 /)

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

3 I) to I 10 00 00 /0 /0to 0 0 to 0/0

4 t r/ I0 10 o 1 /0 t01 10o/ 0 /0(0

5 ID 1 10 10 /0 10 10 10 (0 (0 10

6 M0 10 10 1 O tj ib Ib JO (0 "0 107

/) lb ( 1 1( ) b 10 0 101 / 0 'o 0 toA = Pan weight (nag)Tray color code:: Li-;,,k ;I

Analyst: -l hi.• 13.(.O I4.7 14.31 ml,.4 1 13.• )i , '3.• o . 3. 10 13.3m i4soDate: _______ ,_____

B = Pan + Larvae weight (ing)Analyst: 3Iq.a. tq.3 iq.3z ZoL&5 1S .'I 2 q.q z . t..,qz Il C J' 703r160 I W.zIDate: oft.27.I-t

C = Larvae weight (mg) = B - AAnls:Hand calculated.•t 5.Z7. S.I .tl 'A1.14.. 6.1%; S.%S SAI• 6.1%0 S..S2 I.."S S.0'6 ,.'1%

Anal yst J±ý

Weight per initial number of larvae (rag)C /Initial number of larvae

Hand calculated.4%1* 4NAnalyst' %Y . l" 0 ..

Average weight per Percent reductioninitial number of from control (%) o. %L'.- ?.L1 0. O.'606 -S1. ".7.larvae (Mg)

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

Comments.

Sage 33of 99


:ETSPage 4 of 6

Species: Pimephales promelasClient: TVA / Sequoyah Nuclear Plant, Outfall 101, Non-treated Date:

Day 100% IntakeY Z AA BB

0 C0 Io to0 10

1 I0 (o to 10

2 1) to 10 10

3 (Q '/0 /0 /0

/D1 /0,/05 /o0 /0 /0 t06 (o 0 10 (0

A =Pan weight (ng)Tray color code:: VAnalyst: M" '14.o 14.11 1'4.24Date: • Of.t2.

B = Pan + Larvae weight (mg)Analyst: M .2. Z0.47 T..7-L1.11Date: oS-z7.. .•.

C = Larvae weight (mg) = B - A

Hand calculated. S.S'" ',.i . .i h otj

Analyst-

Weight per Initial number of larvae (mg)= C / Initial number of larvae

Hand calculated. "

Average weight per Percent reductioninitial number of from control (%) 0 •-96 0A 7.larvae (mg) __

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

Comments:

* ge 34 of 99

SOP AT20 - Exhibit AT2O.3, revision 07-01-12

,E

Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101Non-treated

August 14-21, 2012

Pimephalespromelas Chronic Whole Effluent Toxicity TestEPA-821-R-02-013, Method 1000.0

Quality ControlVerification of Data Entry, Calculations, and Statistical Analyses

Project n umber. 8207,._ _ _ _ _ Not fnr Compliance Assessment. Interal Laboratoru OC

Coeatratia(%) Replickte nlitnambered Flnatoumberftla,-ae APa welght(mg) BPaa+Larvae Larvae wetghtrt(m) Wdght /Surviitng Mea iget I Coeffilaetntuarraia Weight/ti-lai mahber MMaa adval Mea weightInlal Cliteataf P-er1a eed.dm f.larvae wseight (.0g) -A-0 nberoftaa (rag) SvMngtumbero" tMa' ea alarva(rg) (*A) 0a--re-rhlauea arao (5u..es ooa rsI (%)

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

A 10 10 14.47 20.30 5.83 0.583 0.583a 10 10 13.58 19.73 6.15 0.615 0.615Control C 10 10 14.24 20.16 5.92 0.592 0.584 5.1 0.592 100.0 0.584 5.1 Not applicable

D 10 10 13.24 18.68 5.44 0.544 0.544

E 10 10 14.10 19.45 5.35 0.535 0.5351"% F 10 10 13.86 20.55 6.69 0.669 0.669 100.0 0.635 10.7 -8.8

G 10 10 13.68 20.54 6.86 0.686 0.686H to 10 13.38 19.87 6.49 0.649 0.649

1 10 10 14.25 20.33 6.08 0.608 0.608J 10 10 12.95 18.54 5.59 0.559 0.559K 10 10 15.67 21.93 626 0.626 0.626L 10 10 14.97 21.83 6.86 0.686 0.686

M 10 10 13.50 19.32 5.82 0.582 0.58243.2% N o0 10 13.60 19.23 5.63 0.563 0.562 9.6 0-563 100.0 0.562 9.6 3.8

0 10 10 14.75 19.62 4.87 0.487P 10 10 14.31 20.45 6.14 0.614 0.614 1

Q• 10 10 12.64 18.78 6.14 0.614 0.614R 10 10 13.87 19.42 5.55 0.555 0.555S 10 10 13.79 19.42 5.63 0.563 0.563T 10 10 .14.52 21.42 6.90 0.690 0.690

U 10 10 13.50 18.82 5.32 0.532 0.532V 10 10 13.70 20.43 6.73 0.6730.7

100% W 10 10 13.37 18.43 5.06 0.506 0.571 12.9 0100.0 0.71 12.9 2.2

X 10 10 14.50 20.21 5.71 0.571 0.571

Y 10 8 13.84 19.21 5.37 0.671 0.537100%Intake Z 10 10 14.78 20.47 5.69 0.569 0.620 11.2 0.56995.0 06 1 .7 -0.4

AA 10 10 14.71 20.22 5.51 0.551BB 10 10 14.24 21.11 1 6.87 060.687 0.6

Outfall 101;Duanett's MSD value:PMSD:

intakeDunnetas MSD.value:PMSD:

0.099417.0

0.072612.4

MSD = Minimum Significant DifferencePMSD - Percent Minimum Significant Difference

PMSD is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Lower PMSD bound determined by USEPA (10th percentile) = 128/6.Upper PMSD bound determined by USEPA (90th percentile) = 30%.

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

USEPA. 2001a. 2001 b. Final Report: Interlaboratoty Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity To. Me hn4,_ Vc zt I • n. 2-Appemii. EPA-t2 t-8-01-004 and EPA-821-B-01-005. US Enviromnental Protection Agency. Cincinnati, OR

File: sqn101_081412data.xlsx

Entered by. J. SumnerReviewed by: G


August 14-21, 2012

Statistical AnalysesaETS6

Larval Fish Growth and Survival Test-7 Day GrowthStart Date: 8114/2012 Test ID: PpFRCR Sample ID: VA I SQN 101End Date: 8/21/2012 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Dlscharge Monitoring ReportSample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelasComments: Non-treated

Conc-% 1 2 3 4D-Cbntrol 0.5830 0.6150 0.5920 0.5440

10.8 0.5350 0.6690 0.6860 0.849021.6 0.6080 0.5590 0.6260 0.686043.2 0.5820 0.5630 0.4870 0.614086.4 0.6140 0.5550 0.5630 0.6900100 0.5320 0.6730 0.5060 0.5710

Intake 0.5370 0.5890 0.5510 0.6870

Transform: Untransformed 1-Tailed IsotonicConc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Mean N-Mean

D-Control 0.5835 1.0000 0.5835 0.5440 0.6150 5.089 4 0.8127 1.000010.8 0.6348 1.0878 0.6348 0.5350 0.6860 10.744 4 -1.242 2.410 0.0994 0.6127 1.000021.8 0.6198 1.0621 0.6198 0.5590 0.6860 8.465 4 -0.879 2.410 0.0994 0.6127 1.000043.2 0.5615 0.9623 0.5615 0.4870 0.6140 9.607 4 0.533 2.410 0.0994 0.5835 0,952486.4 0.6055 1.0377 0.8055 0.5550 0.8900 10.256 4 -0.533 2.410 0.0994 0.5835 0.9524100 0.5705 0.9777 0.5705 0.5080 0.6730 12.860 4 0.315 2.410 0.0994 0.5705 0.9312

Intake 0.5860 1.0043 0.5860 0.5370 0.6870 11.706 4

Auxiliary Tests Statistic Critical Skew KurtStlapiro-Witk's Test Indicates normal distribution (p > 0.01) 0.98195 0.884 0.03805 -0.4693Bartletts Test Indicates equal variances (p = 0.82) 2.21405 15.0863Hypothesis Test (1-tall, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob dfDunnetts Test 100 >100 1 0.09943 0.17041 0.00332 0.0034 0.4689 5,18Treatments vs D-Control

Linear Interpolation (200 Resamples)Point % SD 95% CL(Exp) SkewIC05 87.934IC10 >100IC15 >100 1.0IC20 >100 0.9lC25 >100 0.8IC40 >100IC50 >1000.7

0.6

, 0.4

0.2

0.10.0 - -

-0.20 50 100 150

Dose %

Dose-Response Plot

UZ.5

0.7

0.8

0.5,0

80.4

0•.3.

0.2

0.11

................. . .................1-tall, 0.05 levelof significance

8m i~4. 6

o - 0 S_

(tage 36 of 99

File: sqn101_08l412dt a.xlsxEntered by. J. EumnerReviewed by: -V

TVA / Sequoyah Nuclear Plant, IntakeNon-treated

August 14-21, 2012

ErrvtlronmentalTestlig Solutlonsý Inc.

Statistical Analyses

Larval Fish Growth and Survival Test-7 Day GrowthStart Date: 811412012 Test ID: PpFRCR Sample ID: TVA / SQN 101 - IntakeEnd Date: 8/21/2012 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring ReportSample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelasComments: Non-treated

Conc-% 1 2 3 4D-Control 0.5830 0.6150 0.5920 0.5440

10.8 0.5350 0.6690 0.6860 0.649021.6 0.6080 0.5590 0.6260 0.686043.2 0.5820 0.5630 0.4870 0.614086.4 0.6140 0.5550 0.5630 0.6900100 0.5320 0.6730 0.5060 0.5710

Intake 0.5370 0.5690 0.5510 0.6870

Transform: Untransformed 1-TailedConc-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD

D-Control 0.5835 1.0000 0.5835 0.5440 0.6150 5.069 410.8 0.6348 1.0878 0.6348 0.5350 0.6860 10.744 421.6 0.6198 1.0621 0.6198 0.5590 0.6860 8.465 443.2 0.5615 0.9623 0.5615 0.4870 0.6140 9.607 486.4 0.6055 1.0377 0.6055 0.5550 0.6900 10.256 4100 0.5705 0.9777 0.5705 0.5060 0.6730 12.860 4

Intake 0.5860 1.0043 0.5860 0.5370 0.6870 11.706 4 -0.067 1.943 0.0726

Auxiliary Tests Statistic Critical Skew KurtShapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.88547 0.749 1.36005 1.98359F-Test indicates equal variances (p = 0.20) 5.37752 47.4683Hypothesis Test (1-tail, 0.05) MSDu MSDp MSB MSE F-Prob dl'HomoscedastictTest indicates no significant differences 0.07258 0.12439 1.2E-05 0.00279 0.94881 1,6Treatments vs D-Control

Dose-Response Plot

0.80.7-

0.6

0.50

0.3

0.2

0.1

0O

--------------------- --------- .. ......------------1-tail, 0.05 level

of significance

8 Nq N

(I age 37 of 99

File: sqn10I_081412dataxlsxEntered by: J. SumnerReviewed by: I F

of 6

Species: Pimephales promelasClient: TVA / Sequovah Nuclear Plant Outfall 101. Non-treated

Daily Chemnsfr:

Date: ot'4,V•.-

Ir

.

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

. . d - t'[ ,D.O. and conductivity measurements onli0 TAnalyst MI

1 I 2 =--4Concentration [Parameter

I nH(S.U.)I DO (ma/L)IConductivity

CONTROL [CONonTR *AlkalinityNon-treated (mg CaCO3/L)

Z Hardnessem CaRCOZ.)'*Temperature (0C)pH (S.U.)DO (meL/

10.8% Conductivity(inmhos/cm)

-7-1 0q7 V14

VA.1% _LS.07.C?- 1 "4!6 1 -1-47

D~EI 7. Si II

*Temperature (M)S U

nH (S.U.)J

DO (mF_/) -7.7

21.6% Conductivity IS(gmnhos/cm) _20 I IS*Temnerature (*0) I_•. • II tuq.

I -- * ~ II I~ -I - - I -

pH (S.U.) -741' 1.z -.1 i7. So 751l-- t II* - "- ,. . _ . ,,

DO (mg/L) -'--I .1. ~43.2% TsConductivity .

(gmhos )*Temperature (0C) "1,"1.1 I ...,.1tLI

U U __________ ---- aPH (S.U.) 7.31l

86.4% Conductivity z6&±mhos c) I6

9.01..

255 7-55

1.51 7.511 .9.0

%-b-74-. ..*Temperature (00) "u4.'! H "LL.I.3h

-L- -1 -- 'V+IDH (S.U- "7.C5 I1 -7.'IZ

- --- I. 4 Z.DO (mFJL)

100%

Conductivity(pinhos/c) ___

*Alkalinity

*Hardness(mng CaCO3IL) i*TR chlorine (mg/L) 0O. 1 E

9.'

_10

o.I'DT 0ISI*Temperature (0C)

PH (S.U.)DO (ma/)2Conductivity(iimhos/cm)

100% *AlkalinityIntake (Ing 03 L)

*Hardness 1(Ing Chlorine (m/L)*TR chlorine (mg/L) 0. to

II I r• -- !

7-9Z. . .. . I ---- -- Ik-----

I St

'0.104

".Il t. II *Te'm". "' U " I " I° 5 II M , S U IInitial Final Initial Final Initial Final

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

SOP AT2O - Exhibit AT2O.3, revision 07.-01-12ý7jage38 of 99

.j~.

ETSSpecCliei

Page 6 of 6

ies: Pimephales promelasnt: TVA / Secuo ah Nuclear Plant. Outfall 101. Non-treated Date: c• t."i'A ..l

Dae 1

.1

III

I

I

I Day(Analyst identified for each day. performed pHi, D.O. and conductivity measurements onlv., 7iB

Analyst F- 4 I6Concen- Parametertraon__

PH (S.U.)DO (mg/L)

I 7.Co 5 I II q.•l'l II II

~1.~ I 1..' II

CONTROLNon-treated

Conductivity 0(iimhos/crn) 80*Alkalinity(Mg CaCO3/L)60*Hardness(-g CaCO3/L)

:31S

-7 St

"310 37.3

II I

*Temperature (0C) %AI. I. 71 IP - I -L•' I -,/Ia.

- I ~** 41. I ~*-. ~

pH (S.U.) /;iJS•'l" 7.41 -7. 45 7-51 "7..S 1. QS 7.3 3

10.8%DO (mg/L) - '6'0____Conductivity 2`65a.. us Z%5*7t*Temperature (*C) 'MA •.1. t IM- !6 14.1) "I, k '1%. 2

i i ---- , - p . .. .. . . .,• " - " -- -•-- . . .PH au.) T -7. -7.57. 1.33 1 .,7 .SL-7.iDO nutagv 7.9 --7 .21.6% Conductivity z st __ _

pH (S.U.) i' "7.£1. 7.~l -7.51 -7.SL I ".C; '.z ":4i

DO (mnA/L'43.2% Conductivity

(gmhos/cm V4m1-2 - -7.4

7.7.5.*Temperature . .

4-nH (S.U.)

7-4.3

DO (mF/L)86.4% Conductivity

O~mhos/cm)

0-O ~ ~ ~ ~ . 11.c . I77 *6"

"14 1.94.-7C .aQQ '7_441 -797 .M

-7.S

I

*TemperatuJre C*C)* 6 S i U -

DR (S.U.)

DO (mR/L)

100%

Conductivity

*Alkalinity(mg CaCO 3IL)*Hardness(Mg CaCO3/L)*TR chlorine (mg/L)*Temperature (*C)

i21l7'~

'Vt--wI

~O. /0

II

I 2•.D-U

100%Intake

Conductivity

*Alkalinity i*Hardness(*ag CaCO3/L)*TR chlorine (mg/&)

"IW .. .a.1 -•-a. V.I.I I•4t.-. . -. I .IIitl Final I Initial F15I I nritial1 Final InI6tial IF Finial*Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1.Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet


i age 39 of 99

(D*

0OSEiTSb EnvironmentalTestig Solution,' Inc.

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

Pimephalespromelas Chronic Whole Effluent Toxicity Test

EPA-821-R-02-013, Method 1000.0

Daily Chemical Analyses

Project number: 8207

Day 6Concentration IParameter I .~ I -- - I

pH (SU)

Control

DO (mg/L)Conductivity (imhos/cmAlkalinity (mg/L CaCO3)Hardness (mg/L CaCO3) __

Initial F Inal Initial I Final Initial I Final I nit7.49 7.49 7.60 745 7.67 757.6F 7.8 7.6 78 7.6 7.

302 ,315 310

WE 62

88W

fl I Vi...l I

A h

_______ enpraur(~)74 246 29 4724 27 28 27 4 4 27 21 247 2.

H(SU) 762 744 744 752 745 752 757 432 5 4 5 5 .5 73

______Temperature (C) 24.7 2446 2509 24.7 24.81 2467 2498 2497 24.8 24.6 24.7 2024.8 24.31PH(SU) 7.62 7.44 7.44 7472 7.5 7.50 7.59 7.43 7.52 7335 7.5215 7.55 7.33

DO (mX/L) 7.7 79 8.0 7.8 7.9 789 790 7.6 7.9 768 80 787.8 8.0210.6% Conductivity (jLmhos/cm) 297__ 294___ 294__ 285__ 285__ 285_298

______Temperature (CC) 24.7 24.4 25.0 2467 24.8 24.6 24.9 24.9 24.8 2456 24.7 2480 24.9 24.4pH (SU) 7.61 7.42 7.45 7507 7.51 7.50 7.59 7.42, 7.51, 7.37 7.51 7.52 7.56 7.34

4.% DO (mxnL) 7.7 78.0 9 8.0 7.8 7.9 7.6 . 7.9 7.6 8.0 7.8 7.8 8.02.% Conductivity (gmhos/cm) 280__ 283__280 279 278 276 282

Temperature CC) 24.7 24.4 25.0 24.6 24.8 24.8 24.9 24.6 24.8 24.5 24.71 24.8 24.9 24.4pH (SU) 7.60 7392 7.45 7.51 7.51 7.48 7.58 7.42 7.451 7.34 7451 7.52 7.53 7.,314

86O4% DO( L) 767 7.8 8.0 7.9 8.0 7.8 8.0 7.6 7.8 7.7 8.0 7.8 7.8 7.94.% Conductivity (IRmhos/cni) 254_255 195 2492 24 241~6 2I5~

_______Temperature CC) 24.7 24.43 25.0 _ 24.6 24.8 24.6 24.9 24.6 24.8 24.7 24.7 2468 24.9 24.5piH (SUY) 7.60 7.42 7.42 7.54 7.53 7.49 7.57 7.49 7.44 7.38 7.44 7.57 7.54 7.32DO (mg[L) 7.7 7.8 8.0 8.1 8.1 7.9 8.0 7.7 7.8 7.7 8.0 78. 7.8 7.9

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

_______Temperature (OC) 24.8 24.4 25.01 24.7 25.0 24.6 25.0 24.8, 2. 24. 7 24. 8 24.7 24.9 24.5HIE (SU) 7.61. 7.42 7.44 -7.50 7.52 7.50 7.58 7.4 7.43 7.39 7.434.4 75 7.33.

DO (mg/L) 7.7 79 80 8.0 8.2 8.0 8.10. 7.9 7.9 8.0 8.7. .0C'onductivity (jnhos/cm) 183 186 185 kJd 182 181 17678

100% Itk Alkalinity (mgtL CaCO3) 6826 70 7Hardness (mg/L CaCO3) 72 74 72Total Residual Chlorine (niL <0.10 .<0.10 <0.10ITemperature CC) 2. 24. 4 24.9 24.7 25.0 24.6 25.0 2. 25.0 24.7847 2. 24.9 24.7

File: sqn101_081412chem.xlsEntered by: J. SumnnerReviewed by: A

** Page 1 of 7

OETS

Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1002.0)Species: Ceriodaphnia dubia

Client: Tennessee Valley AuthorityFacility: Sequoyah Nuclear PlantNPDES #: TN0026450Project #: &,Ldl

County: HamiltonOutfall: 101

Dilution preparation information: Comments:Dilution prep (%) 10.8 21.6 43.2 86.4 I100Eftluent volume (mL) 270 540 1080 2160 2500

Diluent volume (mL) 2230 1960 1420 3406 0

Total volume (mL) 2500 2500 2500 2500 2500

Test organism source information: Test information:Oranism age: < 24-hours old Randomizing template color. 6'0 $.bDate and times organisms were born ol't " i2..O- 04o0 lb C Incubator number and shelfbetween: location: 7-6a-Culture board: a2 It r. locaton

Replicate mnuber 1 2 3 4 1S- 6 17 8 9 10

Culture board cup number [. I -W I U M Mt YWT1batch: 1.UTransfer vessel information: PH = -611 S.U. Temperature = L C Seleasm batch:Average transfer volume (mL): 05 .S 6.

Daily renewal information:

Day Date Test initiation and feeding, MHSW Sample numbers used Analystrenewal and feeding, or batch used Outfall 101 Intake

termination time0 ot.N-r IN? O -I - t .rLo041 %2 tOO it .Q_.

1 ~ ~ ~ 1S ___ _________ 2ri~r I2.oels.0% Xtt~

2 o1.n..0' O40 oIq'. 1T- tjOtt. IU t lo t. I It

4 _____. _ _ _ _ Os.lo. I"L 17_.oeI1.QJ I" .IS Itt S 1

doI1.11. iOS"_ 1_9 11-__I,- 'L_ a % . 091t-l.ot L. 60 1

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

7 Of" Lt-1-2- h I

Control information: • Summary of test endpoinils:Control-1 Control-2 Acceptance criteria

% of Male Adults: 07- 07, < 20% 7-day LC5s >00Q1.% Adults having 3'( Broods: 1001 fool, > Ž80% NOEC 1001,,% Mortality: 17. 07. < 20% LOEC >1861.Mean Offspring/Female: 31.7. 1 ,31.3 15.0 offspring/female ChV > 10.,% CV: (,'A"27. I S.6#7. < 40.0 % IIC25 ') 100 1J.

( age 41 of 99

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

:ETSSpecies: Ceriodaphnlia dubtaClient: TVA / Seauovah Nuclear Plant, Outfall 101CONTROL-1 Surv.

Page 2 of 7

Date: __ _ __ _ -I_ _

ival and Reproduction Data

Replicate numberDay 1 2 3 4 5 6 7 8 9 110

1 Young produced 0 C) C D 0 0 1Adult mortality \- k.. %- I %-.

2 Young produced l 0 1 1 1 I- " -Adult mortality J ... [ - le _ -I-

3 Young produced 0 I C_ Adultmortality I - _ -1 I - U I - f --4 Young produced C., 1 ,[ O__ ._ .

Adult mortality [ U•5 Young produced %0i 1 •. _ I .

Final Adult Mortali ty _ _ .- t.. I .Xfor3 u Broods _ 1 0- 0 0 0 0 O INote: MeAdult morta litve D = dead), SB = epit brood (single brood %plit between two days), CO -cariy oere (offcpnng earied over with adult durig trasfer).

tenOfping/emale: _',LCONg: 10.8% Survival and Reproduction Data

1 ______ _______ Replicate number________ ___Day produce 2 3 4 6 7 8 9 01JYoung prd__ • __[• • •O • • • •_. .

Adult mortality _ _•21[Young produced 0 0 0I QI j CL 10 C

JAdult mortality VwV •IL.tL _UJ . j.._ .- ... 1L • U ...] .

No te:ng rdueAdult mortalit(L= fiv .D"- ead , B spli bro snl ro pltbtentody), CO - carr over (f(n care vrwt dl uigtase)

4 Youn produced _______ O..c. 0 0 0J Adult mortality M_ .rai "_ __ I _

Young produced 0N 3 0 C .

Adult mortality __ .. jZ[__ 1. I. C2 Young produced C)6 0t 1 a.i '_ IAdult mortality I -C]~ Lj\- ~C U I _

6 Young produced n 0) 0 .0 .c.10 m F i__fAdult nortality .I 2 L- I~P1_~~~H . L- tL4k

4 Young produced -6 tre T It4 IAdult___ mo talityt.. t

Totalyoung produced • __ 3 S J S .I IL

Final Adult Mortality .U - •. T U.. - _ _

Finot Adult Mortality \ C - - -eCo L.- Qr o •-Note: Mdull mortality (L - live. D - dead), SB -eplit brood (single brood split betwoeen two days), CO -cam over (offispijer caruied over with adult during rasfr

Concentration: i

I age 42 of 99

% Mortality:Mean Offspring/Female: S'1% Reduction from Control-1: -. O7

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

0ETS

Page 3 of 7

Species: Ceriodaphnia dubiaClient: TVA / Sequovah Nuclear Plant. Outfall 101CONC: 21.6% Suri'

Date: ______I__"__L_

ival and Reproduction DataReplicate number

Day 1 2 3 4 5! 6 7 8 9 101 Young produced 0 C_ 1f 0 0 1 0) 0 _

2 Young produced 0 0 0____JAdult mortality U . .L.__3J*Young produced W 0 0 0

Adult mortality I I L-I'.--14 Young produced .S S S. w a I I -

SAdult mortality RE___ Yonpone I •'- I I ' •- -I___ ___ '- I t'- I_'l & [ ' -,

5 Youngproduced I I . [Adult mortality __ - - - -I' 'I UI'

6 Youug produced 1 1C) 0 0 C) I cc > 7_..Adult mortality I U _

7] Young produced 40 1-15 ~~' '

Total young produced -' • • I " - ' 3 I I ? -

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

Concentration:

% Mortality: 07.

Mean Offspring/Female: ,32, 1% Reduction from Control-1: ". |7.

CONw: 43.2% Survival and Reproduction DataReplicate number

Day 1 1 2 3 4 5 6 7 8 9 101 edYoung produce 0 ) ( • ( :

].Adult mortality [12. 7&7 1. U . L.. I ... _ L.. =. LL.. .

Adult mortal'yit - ---2 Young produced I C .. Q 1 5 1 .0 1 1

4 on rdcdAdult mortality I ,. L L. I_ --. I t._ t I I..2 Young produced I1 | () I C I Cz

Adult mortality L L4 Young produced SC) kA 10 S1 6...oJ~ 1 So.I 4C S~..__[Adult mortality C_ C...I ~ I .... ' '- '

4 Yung produced %I L

Adult mortality L . L __ UU.

6 • Young produced 1 ) C) I ..

Adult mortality ___- . - . .- - L- 1 A-f --/7] Young produced 1 | 1

Total young produced -.b Iq -&- 1311 31 a% 1 :31 13'4 13" 1 ask_ L.. •- L_ K_ I _ C .I_ L_

Notef% -_ .4 - - _/ UIUSI.ffWUPt JUIUY. SUU)J. -A - an - k flIý i - V~ - O - Ot OU.'. H

I Concentration: -1% Mortality:Mean Offspring/Female: 3.% Reduction from Control-I: -* 7.

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

(Iage 43 of 99

A!i-

0S*ETSa0

Page 4 of 7

Species: Ceriodaphnia dubiaClient: TVA / Seauoyah Nuclear Plant Outfall 101

CONC: 86.4% Surv,Date: 0'1 _ 1+_17._

ival and Reproduction Data1 Replicate numberDay 1 2 3 4 T 6 17 18 9 10I Young produced C )0 C ) C_J Adult mortality Fk 4_ L, 0 LJ L• L_ [

1 Youngproduced A1 , r.. , 0,"I 0 Q I 0 QAdult modaity \._I \_, =T T. I ° =

3 EYoung produced 1 01 D.Q I .. c>. o .....Q.Adult mortalityI , J L.. I_ ,_ ,

4 1 Young produced

I .utmortality _L7 Young produced i,_,,\I _ 1" ____ 'J I'-1 'LAdult mortality I--I N" I I " %. 1

6 Young produced C)I Adult mortality i [I' I° I1 -I a I-7 Youngproduced IS %4% 14 lt0 %

Total young produced "A S4 6 .- ""S ,• . X1

Final Adult Mortality '- '- --1 \- \_ C INote: Adult mortality (L - live, D- dead), SB -split brood (single brood split between two days), CO - can over (offspri carried over with adult dourim transfer.

Concentration:

% Mortality: 7.Mean Offspring/Female:% Reduction from Control-1: -I .S7.

CONC: 100% Survival and Reproduction DataReplicate number '_"

Day _ 1 2 3 4 5 6 7 8 9 10Young produced1 ___

Adult mortality 1-2 _ I' L_ I.'_' L I L_ - L- IL--2 Young produced C) c))Adult mortality L _. _[0 [ 10 I I o3 AYunglproduiedt 1 L3 C) 1 0 1 [ _ 1 -0 1 - 1

Adult mortality I_ I- I L_ I L_ • I•-4. Young producedAdult mortaliyI I V[ '1 1 '- J _l __I

5 IYoungproducedI- I I VWI Cm J1 ('Adult mortality I 1_ x-I x---I .-- I l I_ -ILI -

6 Young produced [ l o0 0_0__ _i 0 .Adult mortality I_ L k L- - I-----

7] Young produced iI 4e I 1 i 1 9, 19 1 -1 "7 .O it 19Total young produced U T 31 3- 1 3 . 3 ,8 36 YI 31 Z 8

Final Adult Mortality "C.._ -c. N. - L_ -L_ C. -. C .•__AW*s: Adult mortality (L i- live, D - dead), Sn - split brood (single brood split between two days). CO = car over (offssrint carred over with adult durint transer).

i

Concentration: i% Mortality:Mean Offspring/Female: -% Reduction from Control- 1: 1.0Z

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

*age 44 of 99

*ETS6 U~''m•TstfDfm t sawl=*M

Page 5 of 7

Species: Cerlodaphnia dubiaClient: TVA / Seauovah Nuclear Plant. Outfall 101CONTROL-2 Surv

Date: O.t'it.ival and Reproduction Data

Replicate numberDay- 1 2 3 4 5 6 7 8 9 110I Young produced

2 -Yugpodcd _ 0 0___ C) C) C

2 Young prdue 1010_oi,..C.[o ..0.1..Adult mortality \L . L..J- .\J_ j.__3 ~F _____ r_ L 0 __4 Young produced d•t 1= _

Adult mortality L_ - - L I i- I -. I5 Young produced •IC) % -'- I___ 10 10 I'•3

_ A dult mortality I~ %. L - . .1_j L_~ -- %[..6 [Young produced 0 T01 0 1

Adult mortality J__7 Youngproduced

Total young produced .• "' • : A".". • • '

Final Adult Mortality JC: I '- ' •. _-- -- I

X for 3Y Broods y. Y. 5 .2-. 5Z 5ZNote: Adult mortality (L - live, D dead). SB = split brood (single brood split between two days), CO - carry over (offspring carried over with adult durin transfer).

I Ccent.ration:

_Mortal__: _I

IMean Offspring/Female:CONC: 100% Intake Survival and Reproduction Data

Replicate numberDay 1 2 3 4 5 6 7 8 9 101 Young produced 1Q5 1 C) I _ (.2 0 0 L "

I Adultmortality L.2 Young produced 0 0

Adult mortality •).. - I _ '[- 1 Lc _ L.

3 Young produced mrai O p 1 _OI O cI.4 IAdult mortality -,Ij_,,.. L I kI I'. _. L_4 Young produced L I I1 SI S1i[S ISI I I

A dolt mortality Q . - L IL - - -

6 • Young produced [ 1 . T! -W I.. I •A__ dolt mortality [ [~I~*I- -I'_~i' _=IL.61 Young produced [C)j1c01TOK1 0 1 ZII0iC

A__ dult mortality ______ .~L L7 Young produced [6 IS 10 1 .~ .. ~ -1

Total young produced 31 3 ac3 3-6 "3, 32. 1 31- 'AqFinal Adult Mortality \_ L .. LZ = -C7 ýNvote: Adult mortality (L -= lte. D dud), Sa - spit brood (tingle blood split between two days), ) -c an over t(oftain carred over with adult duttna transfer).

Concentration:% Mortality: 07.Mean Offspring/Female:% Reduction from Control-2: - I

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

I age 45 of 99

(ia!

(0

(0

0:T

a E&trn*"nsrtTeUng SIWUmInc,. c


Verification of Ceriodaphnia Reproduction Totals

Control-1

Day Re plicate number TotalD 1 2 03 4 0 6 0 7 0 9 102 0 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 04 5 4 4 5 5 5 5 6 4 6 495 111 11 10 "12 10 9 12 11 11 11 108

L6 0 0 0 0 0 0 0 0 M 0 07 14 18 15 16 16 14 17 15 15 15 155

Total 30 33 233 3 31 28 34 32 30 32 312

10.8%DyRe plicate number Total

1 2 3 4 5 6 0 7 0 0 9 101 0 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 04 L5 6 4 4 3 6 6 4 L6 5 5 3 511 13 12 11 12 12 10 1 12 12 12 1 19

3 0 0 0 0 0 0 0 0 0 0' 0 10 00 0 0 0 o0o0o 0 0

7 18 16 18 17 17 19 17 16 14 15 ,167

Total 36 34 33 31 30 37 34 33 32 30 337

21.6%Day ~Re plicate numberToaDy 1 2 3 4 5 6 17 18 19 110 Toa

1 0 0 0 0 0 0 0 0 0 0 02 0 .0- 0 0 0 0 0 50 0 0 03 0 0 0 0 0 0 0 0 0 0 04 5 5 5 5 6 5 5 4 4 4 485 13 !11 12 10 10 13 13 13 11 13 1196 10 10 0 0 0 0 0 0 0 0 07 118 1 15 F6 16 14 19 17 1-6 1-7 1 6

Total 136 131 33 131 30 137 _L5 _L3 32 130 1328

43.2%

DyRe plicate number Total

S 1 0 0 0 0- 0 0 0 0 0 10 0

2 0 0 0 0 0 0 0 0 0 0 03 0 0- 0 0 0 0 0 0 0 0 0

4" 1 2 3 4 5 1 6 7 1 85 1 1 5 05 13 12 12 12 11 11 12 14 12 12 121

6 0 0 0 0 0 0 0 0 0 0 07 18 17 17 15 15 15 19 15 18 18 6

Tot-s-, M -37-1 541 M i M13131 371= 413r] 31133s

86.4%Replicate number Total

Day 1 2 3 4 _5 6 7 8 9 101 0 0- 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 0 03 0 0 .0 0 0 0 0 0 0 0 04 5 7 5 5 4 5 6 5 6 6 54

S 13 12 12 11 12 12 14 1L2 13 13 124

6 0 0 0 0 0 0 0 0 0 0 07 16 15 19 18 19 16 14 18 17 18 170

Total 34 34 36 34 35 33 34 35 36 37 348

100%

Replicate number TotalDay 1 2 3 4 5 6 7 8 9 10 T

0 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 0 0

3 0 0 0 0 0 0 0 0 0 0 0

7 1818 19 16 15+19 17 20 18 19 179Total 38 37 37 34 32 38 35 39 37 38 365

Control-2

TalRe D3icate number TotalS 1 2_ 3 4 5 1 6 7 8 19 1 101 0 1 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 04 4 5- 5 5 6 4 4 4 4 4 455 10 12 13 12 10 10 10 13 11 12 1136 0 0 0 0 0 0 0 0 0 0 07 16 17 16 114 16 15 118 14 14 15 155

Total 30 34 134 131 132 129 132 131 129 131 313A"

100% Intake

DyRepDlicate number TotalIa 2_ _ 4 5 6 7 18 19 1101

1 0 0 0 0 0 0 0 0 0 0 02 0 0L 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 0-4 6 4 6 5 5 7 5 5 4 6 535 11 12 12 r13 10 14 12 12 13 11 1206 0 0 I0 0 0 0 0 0 0. 0 07 1_6 p C-• 15 1 1 US 1• 15 M 16 S 1s 19 1 17 1 159

Total 1I3 3 313 213 41 33

File: sqnlO1_O81412data.xlsxEntered by. J. SumnerReviewed by V

0



August 14-21, 2012

Ceriodaphnia dubia Chronic Whole Effluent Toxicity TestEPA-821-R-02-013, Method 1002.0



Concentration Replicate number Survival Average reproduction Coefficient of Percent reduction from

(%) 1 2 3 4 5 6 7 8 9 10 (%) (offspring/female) variation (%) control (%)

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

10.8% 36 34 33 33 35 35 34 34 31 32 100 33.7 4.4 -8.0

21.6% 36 31 33 31 30 37 35 33 32 30 100 32.8 7.6 -5.1

43.2% 37 34 34 31 31 31 37 34 34 35 100 33.8 6.7 -8.3

86.4% 34 34 36 34 35 33 34 35 36 37 100 34.8 3.5 -11.5

100% 38 37 37 34 32 38 35 39 37 38 100 36.5 6.0 -17.0

Control - 2 30 34 34 31 32 29 32 31 29 31 100 31.3 5.6 Not applicable

100% Intake 33 29 33 33 33 36 33 32 36 34 100 33.2 6.0 -6.1

Outfall 101:Dunnett's MSD value:PMSD:

Intake:

Dunnett's MSD value:

PMSD:

2.0216.5

1.459

4.7

MSD = Minimum Significant DifferencePMSD = Percent Minimum Significant Difference

PMSD is a measure oftest precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statisticallysignificant in a whole effluent toxicity test.

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

Upper PMSD bound determined by USEPA (90"h percentile) = 47%.Lower and upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET InterlaboratoryVariability Study (USEPA, 2001a; USEPA, 2001 b).

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

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

File: sqn101_081412data.xlsxTable populated from associated "Verification of Ceriodaphnia Reproduction Totals" spreadsheet.

Spreadsheet entered by: J. SurpnerReviewed by: 7


August 14-21, 2012

:ET Statistical Analyses

Ceriodaphnla Survival and Reproduction Test-ReproductionStart Date: 811412012 Test ID: CdFRCR Sample ID: TVA / SQN 101End Date: 8/21/2012 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring ReportSample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Ceriodaphnla dubiaComments: Non-treated

Conc-% 1 2 3 4 6 6 7 8 9 10Control-1 30.000 33.000 29.000 33.000 31.000 28.000 34.000 32.000 30.000 32.000Control-2 30.000 34.000 34.000 31.000 32.000 29.000 32.000 31.000 29.000 31.000

10.8 36.000 34.000 33.000 33.000 35.000 35.000 34.000 34.000 31.000 32.00021.6 36.000 31.000 33.000 31.000 30.000 37.000 35.000 33.000 32.000 30.00043.2 37.000 34.000 34.000 31.000 31.000 31.000 37.000 34.000 34.000 35.00086.4 34.000 34.000 36.000 34.000 35.000 33.000 34.000 35.000 36.000 37.000100 38.000 37.000 37.000 34.000 32.000 38.000 35.000 39.000 37.000 38.000

Intake 33.000 29.000 33.000 33.000 33.000 36.000 33.000 32.000 36.000 34.000

Transform: Untransformed Rank 1-Tailed IsotonicConc-% Mean N-Mean Mean Min Max CVQ/% N Sum Critical Mean N-Mean

Control-1 31.200 0.9968 31.200 28.000 34.000 6.193 10 33.800 1.0000Control-2 31.300 1.0000 31.300 29.000 34.000 5.645 10

10.8 33.700 1.0767 33.700 31.000 36.000 4.435 10 140.00 75.00 33.800 1.000021.6 32.800 1.0479 32.800 30.000 37.000 7.578 10 122.00 75.00 33.800 1.000043.2 33.800 1.0799 33.800 31.000 37.000 6.660 10 136.50 75.00 33.800 1.000086.4 34.800 1.1118 34.800 33.000 37.000 3.532 10 151.00 75.00 33.800 1.0000100 36.500 1.1861 36.500 32.000 39.000 5.954 10 150.50 75.00 33.800 1.0000

Intake 33.200 1.0807 33.200 29.000 36.000 5.991 10

Auxillary Tests Statistic Critical Skew KurtKolmogorov D Test Indicates non-normal distribution (p <= 0.05) 0.98421 0.895 -0.0927 -0.4597Bartiletts Test indicates equal variances (p = 0.35) 6.58769 15.0863The control means are not significantly different (p = 0.91) 0.12078 2.10092Hypothesis Test (1-tall 0.06) NOEC LOEC ChV TUSteel's Many-One Rank Test 100 >100 1Treatments vs Control-1

Linear Interpolation (200 Resamples)Point % SD 96% CL SkewICO5 >100IC10 >100IC15 >100 1.0iC20 >100 0.9IC25 >100 0.8IC40 >100 0.7IC50 >100 0.6

0.5c0.410.3

0.20.10.0

-0.1 - --0.2

-0.310 50 100 150

Dose %

Dose-Response Plot

45..

40

35

330

S25.

S20

= 15,

10l

5.

u . ...-

gN,

8

Cil C;o~ - Co

(tage 48 of 99File zqn101 081412dmetxte

Entered by. , SJ terReviewed by. -- F

TVA / Sequoyah Nuclear Plant, Outfall 101. Non-treated

August 14-21, 2012

0.:T

SEnvinonn,.ntal Tentlfg Solutions. Ins.


Analysis used for PMSD calculation only.

Ceriodaphnia Survival and Reproduction Test-ReproductionStart Date: 8/14/2012 Test ID: CdFRCR Sample ID: TVA/SQN 101End Date: 8/21/2012 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring ReportSample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Ceriodaphnia dubiaComments: Non-treated


10.8 36.000 34.000 33.000 33.000 35.000 35.000 34.000 34.000 31.000 32.00021.6 36.000 31.000 33.000 31.000 30.000 37.000 35.000 33.000 32.000 30.00043.2 37.000 34.000 34.000 31.000 31.000 31.000 37.000 34.000 34.000 35.00086.4 34.000 34.000 36.000 34.000 35.000 33.000 34.000 35.000 36.000 37.000100 38.000 37.000 37.000 34.000 32.000 38.000 35.000 39.000 37.000 38.000

Intake 33.000 29.000 33.000 33.000 33.000 36.000 33.000 32.000 36.000 34.000


Control-1 31.200 0.9968 31.200 28.000 34.000 6.193 10Control-2 31.300 1.0000 31.300 29.000 34.000 5.645 10

10.8 33.700 1.0767 33.700 31.000 36.000 4.435 10 -2.828 *2.287 2.02121.6 32.800 1.0479 32.800 30.000 37.000 7.578 10 -1.810 2.287 2.02143.2 33:800 1.0799 33.800 31.000 37.000 6.660 10 -2.941 2.287 2.02186.4 34.800 1.1118 34.800 33.000 37.000 3.532 10 -4.072 2.287 2.021100 36.500 1.1661 36.500 32.000 39.000 5.954 10 -5.995 2.287 2.021

Intake 33.200 1.0607 33.200 29.000 36.000 5.991 10

Auxiliary Tests Statistic Critical Skew KurtKolmogorov D Test indicates non-normal distribution (p <= 0.05) 0.98421 0.895 -0.0927 -0.4597Bartlett's Test indicates equal variances (p = 0.35) 5.58769 15.0863The control means are not significantly different (p = 0.91) 0.12078 2.10092Hypothesis Test (1-tall, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob dfDunnett's Test 100 >100 1 2.02145 0.06479 32.12 3.90741 7.9E-06 5, 54Treatments vs Control-1

I age 49 of 99

File: sqnlOl 081412data.:dsxEntered by: J. SumpnerReviewed by: --•7

TVA / Sequoyah Nuclear Plant, IntakeNon-treated

August 14-21, 2012

a ETS Statistical Analyses

• Ceriodaphnia Survival and Reproduction Test-ReproductionStart Date: 8/14/2012 Test ID: CdFRCR Sample ID: TVA / SQN 101 - IntakeEnd Date: 8/2112012 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring ReportSample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Ceriodaphnia dubiaComments: Non-treated


10.8 36.000 34.000 33.000 33.000 35.000 35.000 34.000 34.000 31.000 32.00021,6 36.000 31.000 33.000 31.000 30.000 37.000 35.000 33.000 32.000 30.00043,2 37.000 34.000 34.000 31.000 31.000 31.000 37.000 34.000 34.000 35.00086A 34.000 34.000 36.000 34.000 35.000 33.000 34.000 35.000 36.000 37.000100 38.000 37.000 37.000 34.000 32.000 38.000 35.000 39.000 37.000 38.000

Intake 33.000 29.000 33.000 33.000 33.000 36.000 33.000 32.000 36.000 34.000


Control-I 31.200 0.9968 31.200 28.000 34.000 6.193 10Control-2 31.300 1.0000 31.300 29.000 34.000 5.645 10

10.8 33.700 1.0767 33.700 31.000 36.000 4.435 1021.6 32.800 1.0479 32.800 30.000 37.000 7.578 1043.2 33.800 1.0799 33.800 31.000 37.000 6.660 1086.4 34.800 1.1118 34.800 33.000 37.000 3.532 10100 36.500 1.1661 36.500 32.000 39.000 5.954 10

Intake 33.200 1.0607 33.200 29.000 36.000 5.991 10 -2.258 1.734 1.459

Auxiliary Tests Statistic Critical Skew KurtShapiro-Wilk's Test indicates normal distribution (p > 0.05) 0.91798 0.905 -0.1715 0.33113F-Test indicates equalvariances (p.= 0.73) 1.2669 6.54109The control means are not significantly different (p = 0.91) 0.12078 2.10092Hypothesis Test (1-tall, 0.05) MSDu MSDp MSB MSE F-Prob dfHomoscedastic t Test indicates no significant differences 1.45886 0.04661 18.05 3.53889 0.03657 1,118Treatments vs Control-2

Dose-Response Plot

'1"

40

35

30

25

L 20

15

10

5

.0

.... ....... ......... ..4 1-tail, 0.05 levelof significance

C?

8

C14 COe4 2J9.G=

*age 50 of 99File: sqnl 01_0814 2data.xlsx

Entered by: J. SumnnerReviewed by: -

*ETS6E~M-MITngSwmonth., Page 6of 7

Species: Ceriodaphnia dubiaClient: TVA / Seauoyah Nuclear Plant, Outfall 101

Dailv Chemistr:

Date: 0 1 t- I+t ..-

I.Day

',performed pH D.O. and conductivity measurements only.)(Analyst identified for each de0 1 I 2

Analyst Ij~ ITS ~ SVConcentration jParameter

pH (S.U.) -?.CC II 7. s 7.143 11 -7-51 1 '7.55 I_ wDO (mg/L)

CONTROL

Conductivity(ginhos/cm)*Alkalinity I(mg CaCr 3/L)*Hardness(Mng CaCO3/L)

'36C 313 1

at. 'a. .0 S

-Y.CIL 07.q4 0 .4 7508 ( E ~46 S .

*Temperature (QC)U

DH (S.U.)DO (me•J -, "7 .II

10.8% Conductivity(gimhos/cm)

.1 0,L. It

-- 7

*TemDerature (00) M-4.1a a ~ - I--*- II - I - II.

.pH (S.U.) ?7.C= -7.qi B 2S7, S17.SiDO (mfiL)

21.6% Conductivity(iunhos/cm)I

-7.17 1 73

-Loa

-LA-1 _15.-1. '7.4 9

SW~*Temperature (PC)S.

oH (S.U.)- a -~-------4I - - - L

43.2%

i-.-.-..--,

DO (mulL) 7.7Conductivitk f Sq(Aihmos/cm) ZL

9.s;1-55

*Temnerature (C• IlA-1 .4 - V.14- - -1 1- L--pH (S.U.) 7.60 -7~ 7.46 ] S

DO~g/) ____ 7A' 7.__

86.4% Conductivity

________ Tempoerature C0C) IN tpH (S.U.) -7.qq 7.47- -1.C3 1.0,DO (mnwU

100%

Conductivity(Iunhos/cm) I*Alkalinity(Mg CaCO3/L) T.*Hardness(MigCaCO 3/Q)1*.________ choie m1 0.1M*Temperature (00)PH (S.U."DO im 7.1

(pinhos/cin) UIS100% *AlkalinityIntake mg

*Hardness

•TR chlorine (mg/L) 0 .00- Temperature CC) 2S.0 . . 15.1 R".I 1.t

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

SOP ATI I - Exhibit ATi 1.2, revision 07-01-12R.C.wbyg~

01 K-rn

ypge 51 of 99

~•

*ETSPage 7 of 7

Species: Ceriodaphnia dubiaClient: TVA / Seauovah Nuclear Plant. Outfall 101 Date: O0.IL 1...•A•I:"• .'.• "

Dae Ot I L

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

V

Analyst I Yvj,,-.11 S Ic31 f3Z. 11 43Concen-

. trationParameter

-IPH (S.U.)DO (mg/L)Conductivity(umhos/cmr)

7A2

CONTROL *Alkalinityt(mg CaCO3/L)*Hardness(mg CaCO3 /L)____

315

7AM

310 -sZ3

*Temperature (QC)•pm

.71Lý t

DH (S.U.) 'JA~.

10.8% Conductivity I:2e 71S 65_ __(iimhoslcm) v -. r

m

*TemDerature (T) -- A .Ve L.4~ I "..m..tPH (s.u.) 7.O7.S? -7.94 -7.5Z _____ ____

DO (mg/L) 7A Ts C21.6% Conductivity -2

(pmhos/cmn) Z______*Temperature (IC) ir3, 'L0 LAt k s I0 -u.. -L f ~ L.

PH (s.u.) , 67.51 .S'- 7.1 7 7. SC 73DO mglL) 7! 7 .0 7. V

43.2% Conductivity(grnhos/cmn) MCA V40 7,4

_____*Temperature (C) it~ 1.' -1 24 tC . . g *zvH (S.U.) '4.F* 1-7.41 1 Z-19 11 -7.S3 1 -1.4 Li 1 7.SS. A 7.93 V -7.22DO (mg/L)

86.4% Conductivity(gmnhos/cm) 07 1 13.01F th 7-4*TeM "erature (C "

i - S

pH (S.U.) 12q' -1.6 -7~ 76-54 t7ZDO (mc/L)

I.

III

00%I.

Conductivity

*Alkalinity

*Hardness

*TR chlorine (mg/L)*Temperature ('C)

a a-

100%Intake

PH (S.U.)DO (migULConductivity(gmhos/mn)*Alkalinlty(mg CaCO3QL*ardness(Mg CaCO0/L)*TR chlorine (mg/L)*Temperature (QC

_ ntil Fna niil Final IInitial Final IInitial 11Finala

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

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

m•

'-"4'-'4-'?

r__ ge 52 of 99


"E T S Ceriodaphnia dubia Chronic Whole Effluent Toxicity Test: 0EPA-821-R-02-013, Method 1002.0

(1 00@

Daily Chemical Analysesto 0EnTs(a Environmental Testing Solutions. Inc. Project number: 8207

Concentration IParameter Da 0 Da7 Day2 Da 3Initial _7 Final IInitial I Final IInitial_ 2 Final 1Initial Ia

.9 -oH (SU)

r

DO (mffIL)Conductivity (pnhoslcm) I .

Control Alkalinity (maLL CaCO)- 9--

3106288

24.9Hardness (mg/L CaCO3)Temperature (C) 24.71 25.11 24.91 25.0 25.21 24.91 25.2

9 - 9*- 9~ -, ~9 .9- ~.9 -- 9- - I - ~9 - I -pH (SU) 7.621 7.441 7.441 ' 7.58 7.451 7.591 7.571 7.58

-- I .. J A ADO Ima/l,• 7-7 7-K K'O KO 70 N11 Nf0 *70

10.8% DO Arm-/T I 77 78 80 so 79 8 1 80 79Conductivity (ILmhos/cm) 2970M 294 1_*Mff= 294 L%ý ý 285 2 8 5Temnerature (0I0 24.71 24.91 24.91 25.2 24.91 25.01 25_oi 25.11 24R 25 •11 24~ 94121 9421 250

pH (SU) 7.62 7.43 7.44 7.59 7.51 7.59 7.59 7.60 7.52 7.54 7.52 7.56 7.55 7.3021.6% DO(mg/L) 77 78 80 81 79 80 79 79 79 81 80 79 7.8 8.2

Conductivity (pnhos/cm) 280 283 4 280 273 272 273 ? _____28

Temperature(70 247 249 249 250 249 250 250 251 248 250 247 248 24.8 25.0pH(SUJ) 7.61 7.44 7.45 7.61 7.51 7.59 7.59 7.58 7.51 7.54 7.51 7.56 7.56. 7.30

4.2% DO(mgL) 7.7 7.9 8.0 8.1. 8.0 8.0 7.9 7.9 7.9 8.1 8.0 7.9 7.8 8.2Conductivity (pmhos/cm) 254 r 255 @ W 255 249 9 248 7r , 246 259Temperature CC) 247 24.8 25.0 24.9 25.0 25.0 25.0 24.8 24.8 24.8 24.7 24.8 24.8 25.3PH (SU) 7.60 7.43 7.45 7.61 7.51 7.58 7.58 7.61 7.45 7.53 7.45 7.55 7.53 7.28

8.% DO (mg/L) 7.6 7.9 8.0 8.2 8.0 .8.0 8.0 7.9 7.8 8.1 8.0 7.9 7.8 8.2Conductivity (pnhos/em) 203 203 194 192 196 196 205Temperature (oC) 248 25.0 2.0 24.9 25.0 25.1 25.1 251 24.8 24.8 247 249 24.8 25.2pH (SU) !7.60 7.44 7.42 7.63 7.53 7.57 7.57 7.61 7.44 7.54 7.44 7.55 7.54 7.29DO (mz/L) 7.7 7.9 806 8.2 8.1 7.9 8.0 7.9 7.8 8.1 8.0 7.9 7.8 8.3Conductivity (wnhos/cm) 183 183 186 181 179 183

100% Alkalinity (mg/L CaCO3) 72Hardness (mg/L CaCO3 ) 72 71 74Total Residual Chlorine U m<0.10 <0.10 <0.10Temperature (00 24.9 25.0 25.0 24.9 25.0 24.9 25.1 24.8 25.0 24.8 24.8 25.0 24.9 25.2pH (SU) 7.61 7.45 7.44 7.64 7.52 7.57 7.58 7.58 7.43 7.56 7.43 7.56 7.54 - 7.30

DO (mg/L) 7.7 8.11 8.0 8.2 8.2 8.0 8.1 8.0 7.9 8.2 8.0 8.0 7.8. 8.2Conductivity (pmhos/cm) 183 .. 186 . 185 182 . 181 176 1% 184

100% Intake Alkalinity (mg/L CaCO3) 68 687Hardness (mg/L CaCO3 ) 72 74 72!I jKesidual Unlorine jmtL) < 0A0 <0.10 . . . l <0.101

Temperature (C) 25.0 24.91 25.01 25.11 251 24.9 2. 24.71 24.91 24.7 24. 24.7 24.94 25.1

File: sqn101_081412chem.xlsEntered by: J. SumnerReviewed by: 4

IIIiII

:ETSa~-~

Page 1 of 6

Chronic Whole Effluent Toxicity Test (EPA-821-R-02-013 Method 1000.0)Species: Pimephales promelas

Client: Tennessee Valley AuthorityFacility: Sequoyah Nuclear PlantNPDES #: TN0020168Project #: g14"I

County: RheaOutfall: 101

Dilution prep aration information: Comments.Dilution prep (%/) 10.8 21.6 43.2 86.4 100 Each concentration was UV-treatedEffluent volume (mL) 270 540 1080 2160 2500 for 2 minutes to remove pathogenicDiluent volume (mL) 2230 1960 1420 340 0 Interferences.

Total volume (mL) 2500 2500 2500 2500 2500

Test organism information: Test information:Organism age: -fz 0, oL. oLb Randomizing template: tDate and times organisms 01 t I l- . 1 C) Incubator number andwere born between: shelf location: _ _ _

Organism source: ATby,. 60 pTCI ., oC 1i3--L Artemia CHM number: f.1 -Drying information for weightdetermination:

Transfer bowl information: pH= 'Wo.tO S.U. Date / Time in oven: vt', n- t'.ooTemperature = "l.. 0C Initial oven temperature: 10C

Average transfer volume: Date / Time out of oven: o0"J.. t•, *0' ""'"~ Final oven temperature: "

Total drying time: -1A 46

Daily feeding and renewal informiation:

Day Date Morning feeding Afternoon feeding Test initiation, Sample numbers used MHSWrenewal, or batchtermination used

Time Analyst Time Analyst Time Analyst Outfall 101 Intake'

0 O-15.101- 1StL co I's%%4 L % r. ilt9.01 to L0IS.I L %-..%I

3 61.11.1-1. co-t) I t'tQ %UlsI?-~t. 16 k 'Loftm. II L 0-1+1E]

4 af-kt" 1L- o•1t .%<, iq% .L . ll . • 170011. 0e 1208T 1±a1.0 '

5 tA d1Si 9 11 A II2091-.O0 I ti20ai.ol6 i OU'1-nan

71

Control information: Acceptance criteria Summary of test endpoints:% Mortality: -<20% 7-day LCso ) I w7.Average weight per initial larvae: . NOEC W 1 7.Average weight per surviving larvae: O. S l >0.25mglarvae LOEC I tOol.

ChV 1*0 1067.IC 2 5 5 ( 1007.

*age 54 of 99

SOP AT2O - Exhibit AT20.3, revision 07-01-12

of 6

Species: Pimephales promelasClient: TVA / Sequoyah Nuclear Plant, Outfail 101, UV-treated Date: o atwi-IM-

Survival and Growth DataDay CONTROL 10.8% 21.6%

,_A B C D E F G H I J K L

01 ) It to to it) 0 t /C0t I t O C to) to0 1 o 10 3 10 U 1O /0 /0 10 /01 t02 I0 10 o to 10 110 /0 /0 /)0 103 /0 b 0 103 to 0 /0 10 10 10 IC 104 1C) IQ C 10 /o (0 10/0 toIC '

to0 /0 . lo 1 o /0 /0_i /00IC 1c M Q6 10 101(0 to/0 /0 (0 /0 10 (0 '0 /07 IlV

/• 10 to, 0 3 10 0 0 to /0' (0A = Pan weight (mg)Tray color code::Analyst: 146 1-- .. s 1-3 t4.4z 4m.v-8 14.7%{, As 3. .4.|4. orp Is.Date: 08.06.1t-

B = Pan + Larvae weight (mg)Analyst:. 1`. I".i6 aq.zs tl.36 .Z 1 2Loo06 2.o.21 Zas57 IS.1 dt Z0.34 |.qs q.5$5 1-113Date: op.z.7. -I

C - Larvae weight (mg) = B - A

Hand calculated. S tO S A 'sC (-1 S.S 6,O .'b% Is N4 A s.'Il S.aIi .0-Analyst: ,_ ___

Weight per initial number of larvae (mg)= C / Initial number of larvae

Hand calculated. .1Analyst:

I

yN

49 $0*

*0

'9 '00*

40'

Average weight per Percent reductioninitial number of from control (0le) . o.6'!S

- U - - U - - U - * -- U -

- tqA7.a i -I

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

(Iage 55 of 99


of 6

Species: Pimephales promelasClient: TVA / Sequoyah Nuclear Plant Outfall 101, UV-treated Date: OI.i'A-VL.

Survival and Growth Data

Day 43.2% 86.4% 100%,_M N 0 P Q R S T U V W, X

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

2 00 10 10 (C 10 (0 10 to i0 to1

[ 0 to I_ Q10 101 0 10 IO ) 1 to 1O 0 to003 I to I to 10O ( to / 0 10 I /0 /0 / ) / 04 I) /0 10 /C) /0 / 0 ItI 10 /0 /0 IC) /I )5 /0 /0 10 /0 10 /0 /0 ,1A /01 /0 /06 to to 10 to to to 1 0t 11 /0 t O1 0

77__ (o 10 t0 1) t 01 1 I• /0 /b /6Q

A = Pan weight (mg)Tray color code:: L -L&,L 1|.-Analyst- _ In 34D I.' 13.• .14 135.77' 15.A 13-1&• 14.8z ,4.3s IL-.3 13.Uf ILI.

Date: _ _ _n_..__-

B = Pan + Larvae weight (mg)Analyst: -MR 7-_.oo 1L63 1_0I I41.041 18_.36 _ _ _._6 _q.47 tq___ ___._ 16.1_ IS - 17._7_Date: o8.zI7


Hand calculated. SA.'L S~iI. 16 t S.t t S.d% S.A- S..z% 4.1t S S60 SL.6 7 2.J, .sc'i

Analyst:

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

Hand calculated. V,%• i e ,Analyst. C - o" v" U" 0"

L Average weight per Percent reductioninitial number of from control (%) o--.)7. .¢OP. q..&T. 0, S .- ).larvae (mg

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

Comments:

(f age 56 of 99

SOP AT20 - Exhibit AT2O.3, revision 07-01-12

of 6

Species: Pimephales promelasClient: TVA / Seciuovah Nuclear Plant. Outfall 101. UV-treated Date: O•.L- 11-M I I I

Day 100% IntakeY Z AA BB

0 to10 10 (

S/002 to ot3 I b t o t o t4 /0 to (0 10

5 1 _)/0 t 06 (0 (0 t0 10

7 / i____ __ _ 10 to, (0

A = Pan weight (71g•Tray color code:: 4Analyst: lit. i.1Date: O_.__ _. _ _

B = Pan + Larvae weight (nag)Analyst- J-r 2 Z .3 "Date: 0O.z7.1z


Hand calculated. (o . L' LAnalyst-

Weight per initial number of larvae (mg)= C./ Initial number of larvae

Hand calculated.

Average weight per I Percent reductioninitial number of from controllarvae (mig)

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

Comments.

(Iage 57 of 99

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

(D


TVA / Sequoyah Nuclear Plant, Outfall 101UV-treated

August 14-21, 2012

Pimephalespromelas Chronic Whole Effluent Toxicity TestEPA-821-R-02-013, Method 1000.0


Project number. 8207Not for Compliance Assessment, Internal Lablratory QC

Conenatratio() Repfle letlalauamberf mI al-mber-aF an. A=P-uw eght(mg) B-Paa+Lar-se Larewelght(mg) Weight/SonIhag Me-aweight/ Coeffictafvarlatlon Weight/lldtd lnumber Meanerrival Merwreghtl/talt CeltTdeletof Peent reductonfmthanlarvae weightnOg) -A-B mumberaftervue(mg) Surtvirgotreof ateklrw -b oPSot tae(eag) (0) humberoflarvee varitdion g e0ovtn (00)

t-r) m5.r ... (%) (ea 00 ee ra=i,,.)

A 10 10 12.88 17.98 5.10 0.510 0.510B 10 10 14.40 19.28 4.88 0.488 0.488Control C 10 10 15.49 21.36 5.87 0.587 0.535 8.3 0.587 100.0 0.53S 8.3 Not applicable

D 10 10 14.63 20.16 5.53 0.553 0.553E 10 10 13.33 20.06 6.73 0.673 0.673

10.8% F 0 10 14.62 20.21 5.59 0.559 0.614 8.0 0.559 100.0 0.614 8.0 -14.9G 10 10 14.26 20.57 6.31 0.631 0.631H 10 10 12.84 18.78 5.94 0.594 0.5941 10 10 14.35 20.34 5.99 0.599 0.5992 10 10 13.19 18.43 5.24 0.524 0.524K 10 10 14.06 19.53 5.47 0.547 6.0.547L 10 10 13.11 18.93 5.82 0.582 0.582M 10 10 14.08 20.00 5.92 0.592 0.592N 10 10 13.47 18.63 5.16 0.516 0.516432% 0 O1 35 90 .80580.561 6.4 058100.0 0.561 6.4 -5.00 10 10 13.53 19.01 5.48 0.548 0.548

P 10 10 13.17 19.05 5.88 0.588 0.588Q 10 10 13.27 18.36 5.09 0.509 0.509

86.4% R 10 10 13.69 18.96 5.27 0.527 0.521 4.5 0.527 975 0.509 &.6 4.8S 10 10 13.96 19.47 5.51 0.551 0.551T 10 9 14.82 19.30 4.48 0.498 0.448U 10 10 14.33 19.83 5.50 0.550 0.550V 10 10 12.73 18.38 5.65 0.565 0.565

100% V ot 27 83 .50550.534 5.8 O5 100.0 0-534 5.8 0.1W 10 10 13.24 18.50 5.26 0.526 0.526X 10 10 12.76 17.70 4.94 0.494 0.494Y 10 10 15.22 21.43 6.21 0.621 0.621

100% Intake Z 0 10 14.61 20.34 5.73 0.573 0.573 100.0 0.581 12.5 -8.7AA 110 14.37 20.84 6.47 0.647 0.647BB 10 10 14.39 19.21 4.82 0.482 10.482

Outfafl 101:Dunnett's MSD value:PMSD:

Intake:Dunuett's MSD value:PMSD:

0.068512.8

0.082615.5

MSD = Minimum Significant DifferencePMSD = Percent Mirinium Significant Difference

PMSD is a mensum of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically significant in a whole effluent toxicity test.

Lower PMSD bound deter-mined by USEPA (I 0th percentile) = 12%.Upper PMSD bound determined by USEPA (90th percentile) = 30%.

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

USEPA. 2001a. 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes I and 2-Appendix. EPA-821-B-01-004 and EPA-821-B-01-005. US Environmental Protection Agency, Cincinnati, O1.

File: sqnl01 081412data-uv xlsx

Entered by: J. SumnerReviewed by:

TVA / Sequoyah Nuclear Plant, Outfall 101UV-treatedE S August 14-21, 2012

* .Statistical Analyses• EFIonIt frgmetlTl•SolutO n sfle.

Larval Fish Growth and Survival Test-7 Day GrowthStart Date: 8/14/2012 Test ID: PpFRCR Sample ID: TVAI SQN 101End Date: 8/2112012 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring ReportSample Date: August 2012 Protocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelasComments: UV-treated

Conc-% 1 2 3 4D-Control 0.5100 0.4880 0.5870 0.5530

10.8 0.6730 0.5590 0.6310 0.594021.6 0.5990 0.5240 0.5470 0.582043.2 0.5920 0.5160 0.5480 0.588086.4 0.5090 0.5270 0.5510 0.4480100 0.5500 0.5650 0.5260 0.4940

Intake 0.6210 0.5730 0.6470 0.4820

Transform: Untransformed 1-Tailed IsotonicCone-% Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD Mean N-Mean

D-Control 0.5345 1.0000 0.5345 0.4880 0.5870 8.269 4 0.5744 1.000010.8 0.6143 1.1492 0.6143 0.5590 0.6730 7.973 4 -2.807 2.410 0.0685 0.5744 1.000021.6 0.5630 1.0533 0.5630 0.5240 0.5990 6.009 4 -1.003 2.410 0.0685 '0.5630 0.980243.2 0.5610 1.0496 0.5610 0.5160 0.5920 6.414 4 -0,933 2.410 0.0685 0.5610 0.976786.4 0.5088 0.9518 0.5088 0.4480 0.5510 8.649 4 0,906 2.410 0.0685 0.5213 0.9075100 -0.5338 0.9986 0.5338 0.4940 0.5650 5.806 4 0,026 2.410 0.0685 0.5213 0.9075

Intake 0.5808 1.0865 0.5808 0.4820 0.6470 12.504 4

Auxiliary Tests Statistic Critical Skew KurtShapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.94963 0.884 -0.1675 -1.1562Bartlett's Test indicates equal variances (p = 0.97) 0.84084 15.0863Hypothesis Test (1-tall, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob dfDunnett'sTest 100 >100 1 0.06848 0.12811 0.00527 0.00161 0.02854 5, 18Treatments vs D-Control

Linear Interpolation (200 Resamples)Point % SD 95% CL(Exp) SkewIC05 59.875ICIO >100IC15 >100 1.0IC20 >100 0.9IC25 >100 0.8IC40 >100 0.7IC50 >100 0.6

0.50.4

0 0.4S0.3

j 0.2

0.10.0

-0.1-0.2

-0.30 50 100 150

Dose %

Dose-Response Plot

0.8

0.6.

0.6 1-tail, 0.05 levelof significance

00.-(:30.3

0.2

0.1

01

V D 0

6 File: sqnl 01081412data-uv.xlsx

Entered by: J. Su erReviewed by:

Sage 59 of 99

TVA / Sequoyah Nuclear Plant, IntakeUV-treated

August 14-21, 2012

0:T

1 21 lfmfra Tetn Sttonj. Inc.


Larval Fish Growth and Survival Test-7 Day GrowthStart Date: 8/14/2012 Test ID: PpFRCR Sample ID: TVA/ SQN 101 - IntakeEnd Date: 8/21/2012 Lab ID: ETS-Envir. Testing Sol. Sample Type: DMR-Discharge Monitoring ReportSample Date: August 2012 Protocol: FWVCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelasComments: UV-treated

Conc-% 1 2 3 4D-Control 0.5100 0.4880 0.5870 0.5530

10.8 0.6730 0.5590 0.6310 0.594021.6 0.5990 0.5240 0.5470 0.582043.2 0.5920 0.5160 0.5480 0.588086.4 0.5090 0.5270 0.5510 0.4480100 0.5500 0.5650 0.5260 0.4940

Intake 0.6210 0.5730 0.6470 0.4820


D-Control 0.5345 1.0000 0.5345 0.4880 0.5870 8.269 410.8 0.6143 1.1492 0.6143 0.5590 0.6730 7.973 421.6 0.5630 1.0533 0.5630 0.5240 0.5990 6.009 443.2 0.5610 1.0496 0.5610 0.5160 0.5920 6.414 486.4 0.5088 0.9518 0.5088 0.4480 0.5510 8.649 4100 0.5338 0.9986 0.5338 0.4940 0.5650 5.806 4

Intake 0.5808 1.0865 0.5808 0.4820 0.6470 12.504 4 -1.088 1.943 0.0826

Auxiliary Tests Statistic Critical Skew KurtShapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.95559 0.749 -0.6317 -0.2536F-Test indicates equal variances (p = 0.44) 2.69933 47.4683Hypothesis Test (1-tail, 0.05) MSDu MSDp MSB MSE F-Prob dfHomoscedastic t Test indicates no significant differences 0.0826 0,15453 0.00428 0.00361 0.31833 1, 6Treatments vs O-Control.

Dose-Response Plot

0.8

0.7

0.6

f;

. . . . . . . ...0.. 1-I - - - - - - - - -tail, 0 .0 5 level0.4 of significance

0.3

0.2

0.1

0co 0 c;

dz

File: sqnI01_081412dEtta-uv.xlsxEntered by: J. Sumner

Reviewed by

age 60 of 99

I*ETSSpecClien

Dai

Page 5 of 6

ies: Pmneyhales promelasit: TVA / Sequoyah Nuclear Plant, Outfall 101. UV-treatedChemnstrv:

Date:

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

Analyst0 I 1 I 2

Concentration [ParameterI DH (S.U.)I DO (me/L)

Conductivity(Ujmhos/cm'lCONTROL

UV-treated*Alkalinity(mg CaCO3/L)*Hardness(Mg CaCO3/L)

I .,v ...... . .... m•________ T emuperatureUL .. *a . IL, 66 11~ wt -1

pH (S.U.) 7I.CC '7'f 7.5a 1S 1CDO (mg/L) __ _ __ 7,_ _ __ 1 7.q

10.8% Conductivity vqzBZIS(pimho sLq 7.4

_______*Temperature C0C) ILAI VA- I 'L -* (pH (S U.) '7.CC 1-747 7.50 IZ CDO (mgfL) V 1 7.4 7

21.6% Conductivity(jimhos/cm) 7-9Zg4 e

______*Temperature (C) -L4 1. '8 _& 11pH (S.U.) 7.4S 7.Llq I -i'Ll l7.C ftQ~~LDO (mg/L)

43.2% Conductivity(nhsfcmn)

*Temnlerature ('C)a - - a

PH (S.U.)

Z52

-8. C3'7.LoDO (mp/L)86.4% Conductivity

(jpmhocm*Temperature (QC)

S S ~~u tv ii~PH ru. IDO (ma/L)

I"

100%

Conductivity(iunhos/em)-Alkalinity(mg CaC0iIL)m *Hardness

S(me CaMO/L)-I

*TR chlorine (mg/L)

?ICZ -q3 -7-6 7..90 ~~7.4579

1'. 0 -S.

?_ .Co7.6 .5(O.I6

I*Temperature (OC)DH (S.U.)DO (ma/L)Conductivity(pimhos/m*Alkalinity(mg CaCOs/L)

100%Intake

*Hrdness" "" '' .""______I (mg CaCO3lL) Ij*TR chlorine (mg/L) -(o . : "*Temperature (Q "t. . . IS 0.0 ... . ! IFinal Initial Final Initial Final

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

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

• SOP AT20 - Exhibit AT20.3, revisd:n07-1

age 61 of 99,•. ____-_._

I

&M-ei•,.. Page 6 of 6

Species: Pbnephales promelasClient: TVA / Seaunah Nuclear Plant. Outfall 101. UV-treated Date: O&lLt.- .

Day(Analyst identified for each day, erformed PK DO. and conductivi measurements only.3 4 5

Concer- Parametertrati0n

pH (S.U.) • -,7 1.S, 7.0 7." . 77. 7

ConductivityCONTROL (jirnhoslcm)UV-treated *Alkalinity

(mg CaC03/L*Hardness(Mg CaCO3/L) •2. IA-. 2.4- i

_________*Temperature (O* *

PH (s.u.) 7i .8 74 .67 - 7.7 C1( .3DO (mnW -7.

II

KV

10.8% Conductivity _ _ _ li .*TemperatureC) I S.O C n. a.. 4.1 "M4PH, U.) . 7. .- ".* _ o___ .. C .3DO (mgpL) 0 -7. 1 ..

21.6% Conductivity(11mhos/cm)2-p s

-_*Temperature (Q) 11. IIs'. o I. IpH(S.U.) -7 54 -. • .DO (mg/L).'8" 7.9 7..I9.0 1.9 7.9...

43.2% Conductivity(gxmbos/cmn) LC-L17

_____*Temperature (OC) Is -L & ~ . .. ~U.0 if'.PH (S.U.) 7. 7.603 .*I 1 7. 7.3DO hlorin1 .mA 7.j.

86.4% Conductivity

*Temperature CC) LS.. I.S__0 2. V.t 7_ S-.0

Conductivity-Ios/cm 167

1% *Alkalinity1 (%mg CaCO3/L 0

*Hardness

• TR chlorine (rag/L)•

*•Temperature (IQ) "I-%. 2LS "...PH (S.U.) 7.47LI i~ 7A.2 sq 7.DO mglL) I 7..0.97. dqConductivity

Whos/cm) 176 (Sol100% *Alkalinity10Intake (mg CaCOs/

*Hardness(Mg CaCO31L)1-*TR chlorine (mg&L) 0

____*Temperature *Q IS. 1.(.f 2q. 11.Z S I I .7 s

Initial Final Initial 11 Final I Initial 11 Final I Initial I Final*Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located on Page 1.Alkalinity, hardness and total residual chlorine performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bcnch sheet

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

SOP AT220- Exhibit AT20.3, revision 07-01-12.\•jP~yage 62 of 99

0

cc Environmental Testing Solutions, Inc.

TVA / Sequoyah Nuclear Plant, Outfall 101 - UV-treatedAugust 14-21, 2012

Pimephalespromelas Chronic Whole Effluent Toxicity Test

EPA-821-R-02-013, Method 1000.0

Daily Chemical Analyses


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

PH (SU) 7.65 7.49 7.50 7.59 7.62 7.53 7.59 7.47 7.56 7.40 7.59 7.61 7.62 7.34DO (rneIL) 7.8 7.8 7.9 7.8 7.9 7.9 8.0 7.8 8.0 7.6 8.1 7.9 7.9 7.8

cnrl Conductivit (Imhos/cm) 302 309 1302 303 299 ,298 929 M31610Cnrl Alkalinity (mglL CaCO 3) 62 .. 61 61-Hardness (mg1L CaCO3) 84 .j84 RE2784

_______Temperature COC) *24.8 247 24.8 24.7 24.8 24.9 24.9 247 24.9 24.8 24.8 24.6 24.9 2.PH (SU) 7.66 7.48 7.50 7.56 7.63 7.52 7.62 7.45 7.58 7.41 7.57 7.57 7.61 7.34

DO (mg/L) 7.9 7.8 7.9 7.8 7.9. 7. 8.0 7.8 8.01 !7.6. 7.9 7.8 7.9 7.80.% Conductivity (Iwnboslcm) 294 . 298 293__ 291__ 285__ 282__ ffV2X_

_______Temperature (90 24.9 24.5 24.8 24.7 24.9 25.0 25.0 24.7 25.0 24.8 24.8 24.7 24.9 24.5PH (S11) 7.66 7.47 7.50 7.52 7.62 7.52 7.63 7.45 7.56 7.46 7.57 7.58 7.61 7.34

2.% DO (mal) 7.9 7.7 7.9 7.9 7.9 7.8 8.0 7.8 7.9 7.8 8.0 7.8 7.9. 7.92.% Conductivity (pmhnos/ci) 281__ 284__ 280_ 277_27_27_28

_______Ternperature CC) 24.9 24.81 24.8 24.7 24.9 -24.8 25.1 24.8. 25.0 24.8 24.9 24.7 25.0 24.6PH(Sim 7.66 7.45 7.49 7.54 7.62 7.52 7.62 7.45 7.55 7.411 7.54 7.58 7.61 7.33

DO (mgtL) 7.9 7.7 7.9 7.9 8.0 7.9 8.0 7.8 7.9 7.8 8.0 7.8 7.9 7.94.% Conductivity (pinhostcm) 255 * ~ 258 .. 257 255__ 246__ 249__ 256__

_______Temperature (*C) 24.9 24.7 24.8 24.7 24.9 24.81 25.1 24.8 25.0 24.6 24.9 24.8 25.0 24.6PH (SU) 7.63 7.43 7.48 7.56 7.59 7.50 7.61 7.45 7.50 7.38 7.49 7.57 7.59 7.33

DO (nmg/L) 7.9 7.7. 7.9 7.9 8.1 7.9 8.1 7.7 8.0 7.5 7.9 7.8 7.9 7.984% Conductivity (ismhos/cm) 204 2JM =07 - 20'7 ~. *'204 197 ___ 98_ -207__

_______Temnperature (0C) 24.9 24.7 24.9 24.7 24.9 24.8 25.1 26 25.0 24.71 24.9 24.8 25.0 24.3PH (SI1) 7.62 7.43 7.48 7.54 7.591 7.49 7.61 7.2 7.50 7.41 7.47 7.57.59 7.31F

DO (mWL 8.0 7.7 7.8 7.9 8.31 7.9 8.1 788.1 7.5 8.0 788.0 7.9Conductivity (gmhos/em) 16188, 18'6 182 178 181 .187 [N M

100% Alkalinity (mg/L CaCO3) 70 .. 70 '70

Hardness (mgfL CaCO3) 72 . . 72 . 72*Total Residual Chlorine (mg/L) <010 <0.10<01

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

DO (mg%) .8.10 7.7 8.0 7.9 8.2 8.0 8.1 7.8 8.1 7.7 8.0 7.9 7.9 8.0Conductivit (Iwhos/cm) 186 -IO 191 184 182 & .YJ M 178 178 189 MWMN

100% Intake Alkalinity (mgfL CaCO3 ) 70' 70 .... 7Hardnesa (mg/L CaCO3) 71 72 7ýTotai Residuai Chiorine 'es.1w 1W.1 <01 <0. 10Temp erature (0) 25.1 2. 250 24.61 25.0 24.9 25.1 24.61 24.9 2.48 2.512.

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

File: sqnl01_081412chem-UV.xlsEntered by: J. SumnerReviewed by: b

.ETSSEtwimnmentBITesting Solutions, Inc.

Page -7

Page I of 3

Time initiated 15 0,

Time completed ictAnalyst ]

Date analyzed Ei.iK.ZZThtrant normalitv and multinlier detennin atlos

Alkalinity (SM 2320 B)Matrix: Water, RIL =1.0 mg CaCO3 /L

Titrate samples topH = 4.5 S.U.

. . . . . . .. a no and T .. . . . .... .. .p11 of Normality Normality (N) of H2SO 4 pH Factor or Multiplier

Deionized Titrant check Begin End Total f (5 ml Na2CO3 x 0.05)/E (N x 50000)/ 100 ml samplewater reference standard ml ml ml = 0.25/E = Nx 500

= 4.5 S.U. number number (E) (acceptable range = 0.0180 - 0.0220)5.1 %0%q~ 494 it oNSo 10119 0.0 ~ o 1o.3

-1k tote: A*-

Laboratory control standard:.Reference standard True value Sample Alkalinity (MV) % RS = MV / TV x 100

number (TV) volume Begin End Total Multiplier (Mg CaCO3/L) (acceptable range(Ing CaCO31L) (ml) ml ml ml =90 to 1!0%)

io3t" 100 100 it. I tl.* V,.9 10.3 9q 1.90,%Duplicate saf pie precision: ______ _____

Sample Alkalinity %RPI) =

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3IL) ((S - D) /[(S+r)g2-) x 100number (ml) ml ml ml (acceptable range=.+ 10%)

Duplicate (B) D

Matrix spike recovery: ....

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

(mg CaCO3IL) (mi) ml ml ml

-fN$I tO37 sO I0L> Z7.7 -$3.S 16.g (o.3 Iio

Sample alkalinity (B) Measured spike value (MV) %. R = MV / SV x 100(mg CaCO3/L) MV = A - B (acceptable range

(mg CaCO3/L) = 75 to 125%)

- '-19 ' ¶ :oL.

Sample measurements:Sample volume Begin End Total Alkalinity

Sample number Sample ID (ml) ml ml ml Multiplier (Ing CaCOL)

00 a •04"AL H lSLO t 30,S 14.S 6.0 103 (OZ.I..t ___ __ .0.0 6.o •.o ___ _______

001-1- GaA4i6L __ G.) 117.3 11.3 1_ ___ _______

"Oe.|mIffl, _• 17.3 LS.3 11.0 1 110

__.___r___, ____ Z1.3 31.t - o.q I U0

GO _ 35.1. 4S.7 C-5 CZ) Go

MAC,- - 'H. I 1 100 0.0 Co G.o 0_

Reviewed by: I IJX Date reviewed: 00t (.

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

.ETS1 Endonmtnjle In

Page -7 t

Page 7. of 3

Analyst I F .Date analyzed Iog.vI.]t

Alkalinity (SM 2320 B)Matrix: Water, RL = 1.0 mg CaCO3/L


Time initiated

Time completed

Titrant normalltv and multiplier determination:pH of Normality Normality (N) of H2S0 4 pH Factor or Multiplier

DI)O"RI"T- -- Ttrant--_ft iýn End Total = (5 ml Na2CO 3 x 0.05)/E = (Nx 50000)/100 ml samplewater reference -standard ml • • _ = 0f,5/E ffi Nx 500

= 4.5 S.U. number number (E) _0220)

Laboratory control standard:.

Reference standard True value Sample Alkalinity (MV) %RS=MV/TVx 100number (TV) volume Begin End Total Multiplier (mg CaCOsIL) (acceptable range

(119 CaC3/L) (ml) ml ml ml = 90 to 110%)

I,, 03- 100 100 7.9 q5 .3 _____

Duplicate sam pie precision:Sample Alkalinity %RPD

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3/L) {(S - D) /l(S+l)/2]) x 100

number _ _ _ _ _ ml ml ml (acceptable rarge = I 10%)

Duplicate (B) D 1,1 3 A 7Z- 72

Matrix spike recovery: -

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

(me CaCO3/L) (mln) mli ml ml

V-15S 1037 too J 3o.9 -3q.o s.Z ) 1io.3 17o

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

(mr CaCO3/L) -75 to 125%)

72 . q.oy.

Sample measurements:Sample volume Begin End Total Alkalinity

Sample number Sample ID (ml) ml Iml ml Multiplier (me CaCO3L)ao0jt1.tO &f/A' ) IS. 0 nu 31.o qZ.4 3.4 ftjO1 . 76

I-LoS .o& 4 ,_ LCLAI 45A 3.5 73

•ot I-of "Tam st~o') I krT .o0 3.3 3.3 7I't_. l -I I. 1_ 3._ G4.6 3.3 7

I____r _. 04 C. 6. to.0 3.'4 70

I7,01i - 1 -GI AQIiba i0 1___ 10.0 11.4 3.4 70

*z~o~1.O 4/13._ 4. 16.1. 3.4 _____

C~MO I~ V_____ O23.,f %I___ 7b

Reviewed by: I [ . 7I Date reviewed: [o et,'l.17-

Page 65 of 99 SOP C6 - Exhibit C6. 1. revision 06-01-11

* ETSEn~viistmenitakiting Sltos n

Page 7Z

Page 3 )f _.

Analyst IDate analyzed [i.1i.1 I

Alkalinity (SM 2320 B)Matrix: Water, RL = 1.0 mg CaCO3/L


Time initiated

Time completed Z ra011.112.Z

T7trant normallhN, and multiplier determination:pH of Normality Normality (N) of H2SO4 pH Factor or Multiplier

"~m---T-itroat. check Begin End Total = (5 ml Na2CO 3 x 0.05)/E = (Nx 50000)l 100 ml samplewater reference -- ai ý ml ml = 0.25/E = N x S0=4.5 S.U. number number. (acceptable range = 0.0180 - 0.0220)

Laborator control standard;Reference standard Truevalue Sample Alkalinity (MV) % RS = MV / TV x 100

number (TV) Volume Begin End Total Multiplier (mg CaCO3/L) (acceptable range(mg CaCO3L) (ml) ml ml ml 90 to 110%)

-X~ss io37 100 10O Z.3.€ C 3.1 %.5 10.3 '"to

Duplicate sam i precision:

Sample Alkalinity [ %RPD=Sambe S V) volume Begin End Total Multiplier (mgCaCO (mg -D)IRS-D)fl) x 100

Matrix spike recovr:____________

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

(rag CaCO3/L) (ml) ml ml ml

&sS to17. 103.5 4 .7 42. e-5 \o. 170

Sample alkalinity (B) Measured spike value (MV) %R=MV/SVxl0(ra1 CaCO3/L) = 75 to 125%)

-70. 100Sample measurements:. .=

1% Sample volume Begin End Total AlkalinitySample number ' Sample ID (mi) ml ml ml Multiplier (mg CaCO/L)

1S)'x RO3 _1_6 4.. 1 3.,4 4t1 o 7o

a. o I f(4Ao) I JoQ o., 1.0 0.9 4

r2L of 1% _ -a I.o __c 3.4lxatR, \ • & • q~q 7.1 3.5 3,'C;

as. ....

Reviewed by: [IZ• I Date reviewed: F06t_,9 .

Page 66 of 99SOP C6 - Exhibit C6. 1. revision 0 6-01-11

AIM

SErnvronmentalTesting Solutions, Inc.

Time initiated& ".

Time completed " .

Total Hardness (SM 2340 C)RL = 1.0 mg CaCOA/L

Analyst [3Date analyzed [o.g.i.1

Tltrant normalitv and multiblier determination:Titrant Normality check Begin End Total Normality (N) of EDTA. pH Factor. or Mutiplier

reference standard ml ml ml = 0.21E = (N x 50000)/.$1). m'inIipM'0le6number number (E) (acceptable range = 0.0180 - 0.0220) -Ni 66.0

r:.K 418 Iiss 10 3 0.0 10o. o. o.ojq iq.c .... __

LaboratorY control standard:

Reference standard True value Sample Hardness (MV) % RS=•V/ TV :100

number (TV) volume Begin End Total Multiplier (iag CaCO3/L) (acceptable range(mag CaCO3/L) (ml) ml ml ml = 90 to 110%)

: )oqS 40 502 l2 .7L!2.7 2- I eli s ,Duplicate sam ple precision:

Sample Hardness %RPI

Sample Sample ID volume Begin End Total Multiplier (mg CaCO3lL) {(S - D) /[(S+D)/21) x 100number (ml) ml ml ml

___ ___ -__ __ % Z 14.- 4-7 S 14. ____ _---------_N

oejLDI~ uplicate (B) 7 7 .D

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

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

9145's 1041 4o W M~7 z3.7 C-S KC~ 3

Sample hardness (B) Measured spike value (MV) %-R=M/SVxl10

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

(Iag CaCO3/L) 75 to 125%)

Sample measurements:

Sample volume Begin End Total Hardtiess

Sample number Sample ID (ml) ml ml ml Multiplier (ag CaCO3L)

TV = ND Blank(should be = 0 mg CaCO3 /L) D 0.0 0.0 0.. ND

-11_, -4 ' _ z3.7- 7-7. '4 L.5 S.I__

)zo~s-I 7.t7.7 .37 ".4 '.7

_______ et.44m PAA ___ 3Zi .4 3.5 1.1 ___

08-m12.i-4 frh%4SIL' 0 _ 33.5 37.c Li.:s 9_

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

o * ____ 4Z.1 '46.'4 L1.3 __.__.114

6.S~.1 1UN-4 h O 5W got_ 0.0 3.7 3.7 ____________

iZO1SIOZ._ _3. 1.3 3. _ __ 71

_______ 1_______ _____7.~ CeI k 1 11 _SI___1-.q_ 741

N•geil6 OlU• is used, sample must be diluted. Reviewed by: I e ] Date reviewed I081+tZInl........ ..... ......-- .

L

I.

EnvrIoirnfeftltTeatlngSolutIans, Inc.

Page 3q

Page IZ of 3

Time initiated [7 CTime completed -Z._

Total Hardness (SM 2340 C)RL = 1.0 mg CaCO3/L

Analyst a

Date analyzed[~ 3 ]Titrant normality and mult~ipier determination:

I Titrant Normality check Begin End Total Normality (N) of EDTA pH Factor or Multiplierreference ldll al i- = 0.2E -= (N x 50000)/ •50 ml samplenumber number rE) (accepta - 0.0220) =Nx 11000 I

Laboratory control standard:

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

(Ing CaCO3/L) (ml) ml ml ml = 90 to 110%)VMS$ 16ki 40 50 11. M 13 .1 K. qC "57 -6. *•

Duplicate sam ple precision:

Sample Hardness %RFDSample Sample ID volume Begin End Total Multiplier (Ing CaCO3/L) {(S - D) /[(S+:D)/2]) x 100number (ml) ml ml ml

s-2otio*i.-I TV SWi wT I d) 1C. 7t 32.... 119 7Z.--Duplicate(B) .'7 D Z-

Matrix spike recover: ....

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

(mg CaCO3sL) I ml mi ml

xj,3,ss 10q S- 4c, 1C.-7 "t-ZA 5.1• 1t.C10

Sample hardness (B) Measured spike value (MV) %R=MV/SVx 10(mag CaCOd/L) MV = A - B (acceptable range

(Ing CaCO3/L) = 75 to 125%)

"/2.9 S.; 01.0

Sample measurements:

Sample volume Begin End Total HardnessSample number Sample ID (ml) ml ml ml Multiplier (ag CaCO 3/L)

TV--N- -- a Cs -11.t, mz.(should be = 0 mg CaCO3/L)

. _0_ _ .3 1-4.. 41.q "5 111111111.-r 7z.

i aLoA.V,01 -TVAS JIOIUU 1 q.q 33.C 3.7 7Z.

I.39tljO 1 2- 3S.C 37.'j . 7%.

_-Z.O__ _O_ -3 _ W. 3 "'f.4 3.7 7Z

_________Z. __ _ '44.c ~413 3.7 1 M

v~a~l1.O¶ 30.0 3S.'? 3.7 ___ _______

vuLOSIa4.0 f AUOV aJ 1____ 3.1 1A LA' 1___t Ll I

is used, sample must be diluted. Reviewed by: " w. Date reviewed q ' I ]SOP C7 - Exhibit C7. 1. revision 06-01-11

~ Envfronme~ii~sigSolutionsj nc.

Page o

Page 3 of 3

Time initiated

Time completed -'I

Total Hardness (SM 2340 C)RL = 1.0 mg CaCO 3/L

Analyst nId 5ili dDate analyzed [I *q IIIIIIITitant normnality and multipler deltermination:

Titrant Normality check Begin End Total Normality (N) of EDTA pH Factor or MultipliererIeffft- 5 ,,i ml ml = 0.2/E = (N x 50000)l 5O ml sample

number number (E) (acceptable range - =Nx 1000 ~1• j

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

(should be = 0 mg CaCOJL)

,1. ,,10.0' F.LLW .& IL. AD -7.7 I1.o 3.3 Iq.C ,_svt-o*% t.0 J/4 -3' 4iI1.6 15.5 'is , ''

P >4jf~l~tiW•t is used, sample must be diluted. Reviewed by: I] Date reviewed -n iII]SOP C7 - Exhibit C7.1. revision 06-01-11

Page qzPage 1 of_ o

'PETS' EnvironmentallbongSo~lgOutionls, Inc.

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

Matrix: Water, RL = 0.10 mg/LMeter: Accumet Model AR25 pH/Ion Meter

Analyst ;T%3Date analyzed Oi. 1 -I.

Calibration:

I(odidereagent: T"Nr 514C

Acid reagent: I tSri I

0.10 img/L - 1.00 mg/L I SlopeReference standard number -of > 1.06 -mg/L, the saCples INote: For samples with a residual chlorine of > 1.0 mg/L, the samples must be diluted to be within the calibration range.

Laboratory control standard:Reference standard True value (TV) Measured value (MV) % RS = MV / TV X 100

number (mg/L) . (mg/L) (acceptable range 90 to 10%):rj~ss Io•-t 0.50 o.Szq log.-

Duplicate sample precision: ..

Sample Sample 11D Sample characteristics Residual chlorine %RPD = [(S - D) I[(S+D)/21) x 100number I(MgL (acceptable range =:i: 10%)

o*'4. E PGLAhjctb S10t1I.. 1d'4, DuplicateD____ 6 8 4V -

Sample measurements: _____

sample Sample 10D Sample characteristics Residual chlorinenumber .__ _ __

Reagent Blank 1.- _. DOI?

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

Laboratory control standard:Reference standard Trevau (TV) IMeasured value (MV) % RS MV/TV x 100

number I mg/L) I(mg/L) (acceptable range = 90 to 110%)12 •.s \ 1064 0.50 0.471 Reviedr. .y ZII,

Reviewed by " •\'" ]

Date reviewed

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

Page -tSPage _ of i

".ETS6 Envlmnmental Testing Solutlons6 Inc.


Matrix: Water, RL = 0.10 mg/LMeter Accumet Model AR25 pH/Ion Meter

Analyst [0Date analyzed 061.lC.

I,odide reagent: F € sic

Acid reagent: [ Jz L slP

Calibration:- .. 0.10 mg/L I 1.00mg/L ] Slope I

rReference standard number -X14SS lOC 1 0€ o -4.ocY 771

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


Reference standard True value (TV) Measured value (MV) %RS=MV/TVx1001number (mg/L) (mg/L) (acceptable range = 90 to 110%0)

CArSs 10CL 0.50 O.53% IoG.q]

Duplicate sample precision:

Sample Sample ID Sample characteristics Residual chlorine %RPD = {(S -D) /[(S+D)/2]) x 100"number (mg/L) (acceptable range = k 10%)

1,70916-91- 5 103"'llIftt S (-6.0017-

Duplicate I -. oC1 S

Sample measurements: .....

Sample Sample ID Sample characteristics Residual chlorinenumber _ _ _ _ _ _

Reagent Blank r 0.00?6

k eLsg..__.___---_*v2A&tlS.. to c..6SQWti. no\ CcAr,clear

lZ 08 .1~ C.. A- CO).(~ co (4 r c(er, pqr4Les L.O.00 t53


Laboratory control standard:Reference standard True value (TV) Measured value (MV) % RS =MV /TV x 100 1

number (nmg/L) (11g/L) (acceptable range = 90 to 110%) IZ.4 1 C.1 0.50 o. cs q•.o%

Reviewed byDate reviewed

SOP C8 - Exhibit C8. 1, revision 06-01-11Page 71 of 99

Page _ of I

"ETS• Env•ronmentalTestlng Solutions, Inc,


Matrix: Water, RL = 0.10 mg/LMeter: Accumet Model AR25 pH/Ion Meter

Analyst a'sDate analyzed o•e.1.lZ

Calibration:

[ Iodide reagent: [I t s7i7cAcid reagent:- [-•ItI-

-0.10 mg/L 1 1.00 mR/L 7 SlopeReference standard number I Irtss 10C4 1 • r.qac.

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

Laboratory control standard:Reference standard True value (TV) Measured value (MV) % RS = MV/TV x 100l

number (mg/L) (mg/L) (acceptable range = 90 to 110% J

Djss 1o0q 0.50 o.SZo i0'.o% JDuplicate sample precision:Sample Sample ID Sample characteristics Residual chlorine %RPD = {(S - D) /[(S+D)/2]) x 100number (acceptable range = h 10%)

Duplicate D 0 '.001 co ---- • .tM

Sample measurements: .....Sample Sample ID Sample characteristics Residual chlorinenumber ,__m__/L_

Reagent Blank " 0o. oo07 1

M000____ fl0 -5 A~ aaTAAbcolor, clce'r _____

12,086~ 18 0 h wo% lh . Poils: -Se1au, Jee.r-f Lo.06 4Lf

j9J%96q 6 ¶'IqA SQ~va 1 tot.003fy

_________ 'L, rtAce flcotorcd,,efe. jPPAJ&S L~c3



Reference standard True value (TV) IMeasured value (MV) %RS=MVITVx 100

number I(mgIL) I(Mg/L) (acceptable range = 90 to 110%)16C4 i 0.50 Oaiqcit.I I

Reviewed byDate reviewed LJA0-LLJ

SOP CS - Exhibit C8. 1, revision 06-01-111Page 72 of 99


Appendix D

Reference Toxicant Test andControl Chart

Page 73 of 99

E

SEnvironmental Testing Solutions, Inc.

Pimephales promelasChronic Reference Toxicant Control Chart

Organism Source: Aquatox, Inc.

1.0 I I I I I I I I ~ I I I

USEPA Control Limits (+ 2 Standard Deviations)I I 1 7--

0.9 F-

0.8

0.7

0.6

.- ...... ........................... k....................................'

0.5 [-

I II I I I I I I I I I I I I I I I I I1.4

1.2

1.0.J

S0.8

>1 0.6

'ý 0.4

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

......................................... ,

1.4

1.2

I I I I I I I I I I I I I I I I I I I I

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

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

............ ........................................................................................... .

........................ ......

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

1.0

0.8

0.6

0.4

Test date

-.------ 7-day IC., = 25% inhibition concentration. An estimation of the concentration of potassium chloride

that would cause a 25% reduction in Pimephales growth for the test population.Central Tendency (mean IC25)

Warning Limits (mean IC25 ± SA.10 or 5A75)

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

Graphs generated from associated excel spreadsheet.Excel spreadsheet entered by: J. SumnerReviewed b. k

ýý age 74 of 99

0 • Environmental Testing Solutions, Inc.

Test number . Test date 7-day IC T cTL Kga) (g/L Kd)


State and USEPA Laboratory

Control Umits Sw Warning ULmits S3.s

CT-2S CT+2S CT- SAs CTa+SA1

Laboratory

Control ULmits SK"S

CT - S. CT + SA..

USEPA

Warning Umits S,..

CT - S•. CT + S&s

USEPA

Control Umits

CT- S&9 CT+ Sa.g

CV

12

3456789

1011121314151617181920

08-09-11 0.6709-13-11 0.6610-04-11 0.7310-11-11 0.7610-18-11 0.6711-08-11 0.7812-08-11 0.7702-07-12 0.7702-14-12 o.7403-06-12 0.6403-06-12 0.6804-12-12 0.7105-08-12 0.7705-15-12 0.7106-05-12 0.7206-12-12 0.7707-10-12 0.7607-17-12 0.7308-07-12 0.7008-14-12 0.81

0.710.710.710.710.710.710.710.710.710.710.710.710.720.710.710.710.710.720.720.73

0.05 0.62 0.80 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.80 0.08 0.62 0.79 0.15 0.56. 0.86 0.27 0.440.05 0.61 0.81 0.09 0.62 0.80 0.15 0.56 0.86 0.27 0.440.05 0.62 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.81 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.61 0.82 0.09 0.63 0.80 0.15 0.56 0.86 0.27 0.440.05 0.62 0.81 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0.440.05 0.62 0.81 0.09 0.63 0.80 0.15 0.57 0.87 0.27 0AS0.05 0.63 0.82 0.09 0.64 0.81 0.15 0.57 0.88 0.28 0.45

0.98 0.32 0.390.98 0.32 0.390.98 0.32 0.390.99 0.32 0.390.98 0.32 0.390.98 0.32 0.390.98 0.32 0.390.98 0.32 0.390.99 0.32 0.390.98 0.32 0.390.98 0.32 0.390.98 0.32 0.390.99 0.32 0.390.99 0.32 0.390.98 0.32 0.390.98 0.32 0.390.98 0.32 0.390.99 0.32 0.390.99 0.32 0.401.00 0.33 0.40

1.03 0.071.03 0.071.03 0.071.03 0.071.03 0.071.03 0.071.03 0.071.03 0.071.04 0.071.03 0.071.03 0.071.03 0.071.04 0.081.04 0.081.03 0.071.03 0.071.03 0.071.04 0.071.04 0.071.05 0.06

Note: 7-d IC. = 7-day 25% inhibition concentration. An estimation of the concentration of potassium chloride that would cause a 25% reduction in Pimephales growth for the test population.CT = Central tendency (mean IC2s).

S = Standard deviation of the IC25values.

Laboratory Control and Warning UmitsLaboratory control and warning limits were established using the standard deviation of the lCm3values corresponding to the 10th and 25th percentile CVs. These ranges are more stringent than the control and warning limitsrecommended by USEPA for the test method and endpoint.

S.sA = Standard deviation corresponding to the 10"' percentile CV. (SAM = 0.12)5a.us = Standard deviation corresponding to the 2 5P percentile CV. (S&2 = 0.21)

USEPA Control and Waming Umlts

SA.7 = Standard deviation corresponding to the 7 5 t percentile CV. (S&., = 0.38)

Sa~ss= Standard deviation corresponding to the 9 0 " percentile CV. (SAga = 0.45)CV= Coefficient of variation of the IC2s values.

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

File: ppktci_081412JdsxtEntered by:. J. SumnerReviewed by. 99-

00


I ".


Precision of Endpoint Measurements;Organism Source: Aquatox, Inc.

-- 1.00

l2 • 0.75

• -0.500 t0

E0.25

I I I I I I I I I I I I I I C ( 0 m I s

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

- I I I I I I I I I I I I I I I I I I I -

.- I I I I I I I I I I I I I I I I I I F --

30C Kentucky Acceptance Limit (< 30.0%)

20' a. 20 :...

30

20 --.W . a-

10USEPA Upper PMSD Bound (90'h percentile < 30.0%)

012 0.. .

10

I I I I I I I I I III I I I I I I I I_.. .................. Test.d.......ate . . ... .. .,......

Test date

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

- - Central Tendency (mean Control Growth, CV, or PMSD)......... Control Limits (mean Control Growth, CV, or PMSD ± 2 Standard Deviations)

ýýage 76 of 99

Graphs generated from assodate1 excel spreadsheet.Excel spreadsheet entered by: J :umnerReviewed by:

Precision of Endpoint Measurements

SPimephales promelasEnvironmentalTestinggSolutions,.In. Chronic Reference Toxicant Data

Test number Test date Control Control Mean cv Cr MSD PMSD CTSurvival Growth

for Control Growth for Control W for PMSDK (mg/larvae) (mg/larvae) N Growth CV (%)

1 08-09-11 100 0.662 6.6 0.07 10.6

2 09-13-11 100 0.848 0.755 6.7 6.7 0.12 13.6 12.13 10-04-11 100 0.768 0.759 16.8 10.0 0.13 16.9 13.74 10-11-11 i00 0.681 0.740 4.3 8.6 0.07 9.9 12.8

5 10-18-11 100 0.693 0.730 4.8 7.9 0.07 10.0 12.2

6 11-08-11 100 0.792 0.741 10.3 8.3 0.12 15.6 12.87 12-06-11 100 0.738 0.740 15.7 9.3 0.12 15.9 13.2

8 02-07-12 100 0.662 0.730 10.5 9.5 0.09 13.4 13.29 02-14-12 97.5 0.658 0.722 5.2 9.0 0.07 10.1 1.2.9

10 03-06-12 100 0.854 0.735 3.5 8.4 0.09 10.8 12.711 03-06-12 100 0.873 0.748 5.1 8.1 0.10 11.4 12.6

12 04-12-12 97.5 0.706 0.744 7.0 8.0 0.06 8.7 12.2

13 05-08-12 100 0.613 0.734 8.1 8.1 0.08 13.1 12.314 05-15-12 100 0.697 0.732 5.9 7.9 0.07 9.5 12.115 06-05-12 97.5 0.596 0.723 3.8 7.6 0.06 9.4 11.9

16 06-12-12 100 0.723 0.723 6.0 7.5 0.11 15.6 12.2

17 07-10-12 100 0.641 0.718 5.2 7.4 0.10 14.9 12.318 07-17-12 100 0.638 0.713 10.9 7.6 0.07 11.4 12.319 08-07-12 97.5 0.660 0.711 6.7 7.5 0.06 8.7 12.1

20 08-14-12 100 0.533 0.702 5.4 7.4 0.07 13.1 12.1

Note: CV = Coefficient of variation for control growth.

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

Upper CV bound determined by USEPA (90td percentile) = 20%

MSD = Minimum Significant Difference

PMSD = Percent Minimum Significant DifferencePMSD is a measure of test precision. The PMSD Is the minimum percent difference between the control and treatment that can bedeclared statistically significant In a whole effluent toxicity test.Lower PMSD bound determined by USEPA (10th percentile) = 12%.Upper PMSD bound determined by USEPA (901P percentile) =30%.

Cr = Central Tendancy (mean Control Growth, CV, or PMSD)

USEPA. 2000. Understanding and Accounting for Method Variability in Whole Effluent Toxicity Applications Under the National Pollutant Discharge Elimination Program.EPA-833-R-00-O03. US Environmental Protection Agency, Cincinnati, OH.USEPA. 2001a, 2001b. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes 1 and 2Appendix. EPA-821-B-01-004 and EPA-821-8-01-005. US Environmental Protection Agency, Cincinnati, OH.

File: ppkclcr_0814]2.xlsxEntered by: J. Sumner

age 77 of 99 Reviewed by: __

*ETSEnv~roarnentalTn~tngSouinIs

Page I of 5

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

Species: Pimephales promelasPpKCICR Test Number: _1.

Dilution preparation information: Comments:KC1 Stock INSS number: 114sý _ _o__

Stock preparation: 50 g KCI/L:Dissolve 50 g KCI in 1-L MilHi-Q water.

Dilution prep (mg/L) 450 600 750 900 1050Stock volume (mL) 9 12 15 18 21Diluent volume (mL) 991 988 985 982 979Total volume (mL) 1000 1000 1000 1000 1000

Test organism information: Test information: _

Organism age: __".oAX_____ 0._3t) Randomizing template:Date and times organisms oa- b. r oiv. o Incubator number andwere born between: shelf location: _ _ _

Organism source: t • , " Artemia CHM number: CJltt L.Drying information for weightdetermination:

Transfer vessel pH = "1 eO* S.U. Date / Time in oven: e6&t- -a- i'tinformation: Temperature = 0,A 0C Initial oven temperature: ItAverage transfer volume: Date / Time out of oven: fL.fl. la

0. 'L "A Final oven temperature: 1 ,Total drying time: mg-A•

Daily feeding and renewal information:

Day Date Morning feeding Afternoon feeding Test initiation, renewal, MHSWor termination batch used

Time Analyst Time Analyst Time Analyst

0 __-_L %So•0. VL.S 090 .0-

02 ,-IS.. I. IMlt 0(3 1•t % A( It'~ ms .) *t- IL.

4 at-I fl. I?.. 01A1 0116 Mh~.5__ 08 -Jr.r lgQ- I- 61c AA~ 4 j... loPat-

6 0110-mi.. O100 Vs oa 117. r% at- 16 I-).

-724 iot, -21

Control information: Acceptance criteria Summary of test endpoints:% Mortality: .. IT. < 20% .7-day LCs &U. 'L(,Average weight per initial larvae:. O.-31 N NOEC 00Average weight per surviving larvae: (. 1 2 ! 0.25 mg/larvae LOEC ":! 0

ChV (016. noIC25 I d1S

(Iage 78 of 99


.ETS(b Envtmnn-r0ITestlng Solutlom In

Page 2 of 5

Species: Pimephales promelas PpKC1CR Test Number: W)$_

Survival and Growth Data

Day Control 450 mg KC[/L 600 m KCI/L-.A B C D. E F G H I J K L

0 101 0 1to It O tO to 0 to I /( O /1 /0

1 10 10 to 1 10to 10 t 10 1.0 £ 0) /0

2 10 tO tO 1O to /o to /O /O to /0 (O

3 (o t ( (t 0 10 to t to 10 10 10 10 (0

4__ _1 10 10 10 10 l. tO 0 qI& 1'A (0 /05 /0 /0 /0 t0 t0 /0 /0 /0 C ' ) O 06 / i U 10 "0 I1 I /0 .1 ) to(0 IO

7 tO_/0 _0 10 'O 1O0 to 10 'o'i to /0A = Pan weight (mg)Tray color code:: 91;t,Analyst:. J• 130•.• 4 i.q 1 48 '. 1 3.3 138 143 14.4 l t12. 1f.91 I a.&Ij I3 ; -&.4-2Date: 0R.o1.t_

B = Pan + Larvae weight (mg)Analyst: 40 1017 19 53 7A46 Iq.-3q 14.66 I'l. Zo.ll V-50 19.50 .o.31 14.40 I9.1ZDate: o 194.7.. " •

C = Larvae weight (rag) = B - A

Hand calculated. S ,4 5. s. ie $ .1 I. ,.E1 AA* q.L 6i.'• S.S S (j.16Analyst:

Weight per Initial number of larvae (mg)= C /Initial number of larvae

Hand calculated.Analyst: 0*

½A,

'V'4

0*

A,'4

C,0

½0

0IV

Average weight per initial Percentnumber of larvae (mag) reduction.

from control 0.%MI_-. -L.7 - , 0 L

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

Comments.

I age 79 of 99

SOP AT21 - Exhibit AT21. 1, revision 06-0 1-11

SS*ETS

Page 3 of 5

Species: Pimephales promelas PpKCICR Test Number-. -IS

Survival and Growth DataDay 750 mi KCIL 900 mg KCI/L 1050 mg KCI/L

M N 0 P Q R S T U V NV X0 /D t0o 0 10 10 to I0 /0 /0 /0 /o /0

4_b _ t' "_ i, " - _"z 2_20 M :~ to 0 1- ; -Qa.~'

3

1C0 0% Ilk l_.

__ __ __ &" '' cj*• - u -i 5- I',x .6

A = Pan weight (mg)Tray color code::Analyst: 13s. [4B17. 1 3.18 1zS 4.-.s•. 13.42 IZ. U 14. it1.05 13 IS. 13.O& 13.1fDate: __ _ __ _ _ ._ _ _

B = Pan + Larvae weight (mg)Analyst: ,,.gj q.'a.. l rTBq f 17. ZS I i7.1O " tq.7• -Date: OfE't,.z


Hand calculated. qLii 4),s S.11. 3.1t. S,'S t*,'s aoA O.•S I|S&Analyst:

Weight per initial number of larvae (mg)= C / Initial number of larvaeHand calculated. _ 4. .O 0Analyst: ... A I' •' "

Average weight per Initial Percentnumber of larvae (mg) reduction .O.U

from control io. -

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

V

Comments:

ge 80 of 99

SOP AT'21 - Exhibit AT21.1, revision 06-01-11

(D

0

"• Environmental Testing Solutions, Inc.

Pimephales promelas Chronic Reference Toxicant Test

EPA-821-R-02-013, Method 1000.0


Test numben 255

Test dates: August 14-21, 2012

cowetrbniIneM/LXM Itp!na MW hnumbrnrerof Reinrmehnof A.P-Wft (nfq 0 Pes-tan Lan-e. rnia5j: Wdlit/Sul•veitn MAffi-W61ht/Sunung. Ca.errM ofnmtirton iuh1tlInffm.nunrbnrof M..rurcta(ll )MuNJ ihti~klfth Co.:Mfetnfvnitticn Per-tredunaonofmI- banta nelet ns A-0 '-abWenfhme. lea) urenrhmolme. ee mtv ane.lief number ofiur. ) mnb)l (%j

-.- 4 m) (%) (-1d

A 10 10 13.63 19.27 5.64 0.564 0.564

Control B 10 10 13.46 18.53 5.07 0.507 0.533 5A 0.507 100.0 0.533 5A Not applicableC 10 10 14.89 20.40 5.51 0.551 0.551D 10 10 14.13 19.24 5.11 0.511 0.511E 10 10 13.88 19.66 5.78 . 0.578 0.578

450 F 10 10 13.43 19.19 5.76 0.576 0.607 5.9 0.576 100.0 0.607 5.9 -13.9G 10 10 14.54 20.81 6.27 0.627 0.627H 10 10 15.02 21.50 6.48 0.648 0.648

.1 10 9 13.81 18.50 4.69 0.521 0.469600 J 10 - 9 14.69 20.32 5.63 0.626 0.586 8.2 0.563 95.0 0.558 11A

K 10 10 13.62 19.40 5.78 0.578 0.578 94.5

L 10 10 1342 19.62 6.20 0.620 0.620

M 10 8 13.12 17.89 4.77 0.596 0.477

750 N 10 9 14.56 19.24 4.68 0.520 0.547 7. 0.468 87.5 0.478 5.2 10.5O 10 9 13.78 18.31 4.53 0.503 0.453P 10 9 12.86 17.98 5.12 03569 0.512

Q 10 5 14.59 17.69 3.10 0.620 0310

900 R 10 6 13.72 17.28 3.56 0-593 0.607 4.4 0.356 57.5 0.348 12.4 34.85 10 7 12.18 16.23 4.05 0.579 0.405T 10 5 14.11 17.30 3.19 0.638 0.319

U 10 1 12.85 13.36 0.51 0.510 0.051

1oso V 10 2 13.13 14.76 1.58 0.790 0.650 30.5 0.158 7.5 0.052 142.6 90.2W 10 0 0.00 0.00 0.00 0.000 0.000X 10 0 0.00 0.00. 0.00 0.000 0.000

Dunnett's MSD value:PMSD:

0.069913.1

MSD = Minimum Significant DifferencePMSD : Percent Minimum Significant Difference

PMSD is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statistically significant In a whole effluent toxcity test.

Lower PMSD bound determined by USEPA (10th percentile) = 12%.

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

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

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

Filme ppkcicr -_081412.xlsxEntered by: J. SumnerReviewed b. _4

:ET Statistical Analyses

Larval FishStart Date: 8/14/2012 Test ID: PpKCICREnd Date: 8/2112012 Lab ID: ETS-EnviiSample Date: Protocol: FWCHR-lComments:Conc-mglL 1 2 3 4

D-Control 1.0000 1.0000 1.0000 1.0000450 1.0000 1.0000 1.0000 1.0000800 0.9000 0.9000 1.0000 1.0000750 0.8000 0.9000 0.9000 0.9000900 0.5000 0.8000 0.7000 0.5000

1050 0.1000 0.2000 0.0000 0.0000

3rowth and Survival Test-7 Day SurvivalSample ID: REF-Ref Toxicant

r. Testing Sol. Sample Type: KCL-Potassium chlorideEPA-821-R-02-013 Test Species: PP-Pimephales promelas

Transform: Arcsin Square RootConc-mg/L Mean N-Mean Mean Min Max CV% N

D-Control 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4450 1.0000 1.0000 1.4120 1.4120 1.4120 0.000 4600 0.9500 0.9500 1.3305 1.2490 1.4120 7.072 4

*750 0.8750 0.8750 1.2138 1.1071 1.2490 5.846 4'900 0.5750 0.5750 0.8820 0.7854 0.9912 11.405 4

*1050 0.0750 0.0750 0.2757 0.1588 0.4636 53.294 4

Rank 1-TailedSum Critical

18.00 10.0014.00 10.0010.00 10.0010.00 10.0010.00 10.00

Number TotalResp Number

0 400 402 405 40

17 4037 40

Auxiliary Tests Statistic Critical Skew KurtShapiro-Wilk's Test indicates normal distribution (p > 0.01) 0.93302 0.884 0.39148 0.28113Equality of variance cannot be confirmedHypothesis Test (1-tall, 0.05) NOEC LOEC ChV TUSteel's Many-One Rank Test 600 750 670.82Treatments vs D-Control

Trimmed Spearman-KarberTrim Level EC50 95% CL

0.0%5.0%

10.0% 902.19 867.55 938.21 1.020.0% 909.49 869.16 951.70

Auto-7.5% 898.26 863.27 934.67 0.90.8

0.7

0.6

0 0.5

0.4

0.3-

0.2

0.1,

0.0,1 10 100 1000 10000

Dose mgIL

Dose-Response Plot

1 1

0.9

0.8

0.7

0.6030.5

S0,4

0.3

0.2

0.1

*age 82 of 99

File: ppkclcr.0814l:t.xlsxEntered by:. . Sumner

Reviewed by: ' .

:ETS0 nl~~?a ~~g.oulkI(Statistical Analyses

Start Date: 8/1412012End Date: 8/21/2012Sample Date:Comments:

Conc-mglL 1 2D-Control 0.5640 0.5070

450 0.5780 0.5760600 0.4690 0.5630750 0.4770 0.4580900 0.3100 0.3560

1050 0.0510 0.1580

Larval Fish Growth and Survival Test-7 Day GrowthTest ID: PpKCICR Sample ID: REF-Ref ToxicantLab ID: ETS-Envir. Testing Sol. Sample Type: KCL-Potasslum chlorideProtocol: FWCHR-EPA-821-R-02-013 Test Species: PP-Pimephales promelas

3 40.5510 0.51100.6270 0.64800.5780 0.62000.4530 0.51200.4050 0.31900.0000 0.0000

Transform: UntransformedConc-mg/L Mean N-Mean Mean Min Max CV% N

D-Control 0.5333 1.0000 0.5333 0.5070 0.5640 5.353 4450 0.6073 1.1388 0.6073 0.5760 0.6480 5.924 4600 0.5575 1.0455 0.5575 0.4890 0.6200 11.433 4750 0.4775 0.8955 0.4775 0.4530 0.5120 5.244 4900 0.3475 0.6517 0.3475 0.3100 0.4050 12.430 4

1050 0.0523 0.0980 0.0523 0.0000 0.1580 142.558 4

1-Tailedt-Stat Critical

-2.307 2.180-0.756 2.180

MSD

0.06990.0699

IsotonicMean N-Mean0.5703 1.00000.5703 1.00000.5575 0.97760.4775 0.83740.3475 0.60940.0523 0.0916

Auxiliary Tests Statistic Critical Skew KurtShapiro-Wilk's Test Indicates normal distribution (p > 0.01) 0.94803 0.805 -0.6553 0.58729Bartlett's Test Indicates equal variances (p = 0.40) 1.85289 9.21035Hypothesis Test (1-tall, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F.Prob dfDunnett's Test 600 >S00 0.06992 0.13112 0.00569 0.00206 0.11571 2,9Treatments vs D-Control

Linear Interpolation (200 Resamples)Point mg/L SD 95% CL(Exp) SkewIC05 629.55 44.05 422.75 677.99 -1.2511IC10 683.02 39.17 471.16 745.11 -1.6458IC15 736.48 32.27 546.98 788.00 -2.5024 1.0IC20 774.58 14.39 725.29 814.40 0.0156 08IC25 807.48 14.81 783.34 852.44 0.1502 0.8IC00 902.72 14.24 850.75 928.37 -0.5101IC50 931 6i9 908R 905.24 9-5694 01-578 0.7-

0.6

0.5

CO 0.4I0.30.2~

500 1000 1500

Dose mg/L

Dose-Response Plot

1.04

0.3

0.2-

1-tall, 0.05 levelof significance

(V age 83 of99

Vile: pplciattl1l2.xIsEntered by: Jl. SuninerRemiwed byr.-

00ITSýEnvlmonment.Tesfn Sltl o I

Page 4 of 5

Species: Pimephales promelas PpKC1CR Test Number: _IS

Daily Chemistry:• Day

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

Concentration Parameter. pH (S.U.5) 77.• Ss.•. •_

D O (m g/L)". 7 . .. . ;" . .. . . " . ... . ' " " . .-Conductivity 0 ""-• .... ".. : . o

CONTROL *Alkalinity(mg CaCO3/L) O-" *Hardness .(mg CaCO 3IL) i '1

,____*Temperature (OCD. VA. A. . it

pH (S.U.) .7.73". .'DO (mg/L) 7.7 . 77 •

450 mg KCI/L Conductivity

*Temperature (IC) "k.4. - . i. SpH (S.U.) 1.•S i.s7 l.o 7-4SZ. 7oIDO (mg/L) 14 7. 7.8

600 mg KCIIL Conductivity_______ mhos/cm) (C 133 %360 bV N

*Temperature (0QD - " . C 7. -3 "__._ 4. -

DO (mg/L) "7.7 "7.0 1750 mg KCIIL Conductivity

(gmhos/cm) 100 1 CAO_________ *Temperature (op 14. 46 T41pH (S.U.) a .5?-4 .C

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

(~mhoslcm) Is1&__ _ __ _ Temperature (0) l'4q -Lq.14. 2. . ..ii .ýq

pH (S.U.) '711 II7c:; 1 -7.-q I ~113 1 : -18 11 14.-- 1DO (mg/L)

1050 mg KCI/L Conductivity(jLmh sc) -T Z O70*Temperature (IC)

STOCK Conductivity -'qo0_ (Rmho

Initial II Final I Initial II Final I Initial 11 Final I\Ix

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

* age 84 of 99


....... OET

E~wlmmeul'uTesung &olIw~lo.In

Page 5 of 5

Species: Pimephales promelas PpKC1CR Test Number: 21K

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

3 . 4 5 :_____Analyst am zru3 S 0 :sf V

Concentration ParamterpH (S.U.) 7.CO 7.,7 V... ., " ".I -DO(mg/L) ... 7.. i i.Ci ..Conductivity(jinhoscm)31

CONTROL *Alkalinity(nig CaCO3/L) 12*Hardness

ing CaCO3/L)*TemperaturepH (S.U.) ' .7.7.I 7.4 - 71o 7,L. . 7.l .. .SDO (mg/L)

450 mng KCI/L Conductivity(l'imhos/cm) O)*Temperature

pH(S.U.) O3.-+ -7.S7 7.70 -.7. .77 -i. itS 7.7. 1DO (mgiL) 7.

50 nmg KCIL Conductivity

-(gmhos/m) 1

*Temperature ..... '" " U . .. .pH (S.U.) -3 Q y. -. 7.77 .73:77 1i I _._7' .DO (mg/L) 7.__ .7.'7

900 mg KCI/L ConductivityhTempe r .,6

*Temperature

______ i•.o -iM. 2.M.1 •L•.I. 0 -Ll.6. -A,. i,.44 •-uA.SpH (S.U.) -a, 7.C-' -i.'iq 7 •Lj 7.1 -7.- 7.7q 7.$DO (mg/L) -7 17.3 7,C 7. 7.

goo0mg KCIIL Conductivity41tO

*Temperaturei"! S A.. L.l. ',' "1.(Oc) 0M -6 4

Initial I Final. I Initial I Final I Initial 11 Final I Initial I Final IU - - U 0, i

*Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily Renewal Information table located qa Page 1.Alkalinity and hardfiess performed by the analyst identified on the bench sheet specific for each analysis and transcribed to this bench sheet by: 7 V

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

Ec

• Environmental Testing Solutions, Inc.

Ceriodaphnia dubiaChronic Reference Toxicant Control Chart

1.14

1.12

1.10 -

1.08

1.06

1.04

1.02

2.5

I I I I I I I I I I I I I | I I I I I

USEPA Control Limits (k 2 Standard Deviations)

I I t II I I I I I I I I I I I I I I I

zJ:

N,

2.0

1.5

1.0

0.5

1.4

1.3

I I I I I I I I I I I I I I I I I I I I

" Laboratory Warning and Control Limits (10th and 2 5 th Percentile CVs)- ............ .. ...... .......*. ..... .......... ......... . . . . . .

1.2 H-

1.1

1.0 -

...... .... 0 .... 7-

0.9

0.8 I II I Ifi fI~ I I~ L I I

INN NN NN NN NN -IN NN INN N'111 NN N$ N$ 0 N$ -'s N, Ns -o WNsOk"', NO-N" OSATest date

7-day IC25 = 25% inhibition concentration. An estimation of the concentration of sodium chloridethat would cause a 25% reduction in Ceriodaphnia reproduction for the test population.Central Tendency (mean IC2s)

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

Control Limits (mean IC25 + SA 25, SA90' or 2 Standard Deviations)

-,. age 86 of 99

Graphs generated fromassociated excel spreadshleet.

Excel spreadsheet entered by: J. SumnerReviewedby

fY

.ETS8 Environmental Testing Solutions, Inc.


Test number Test date 7<day IC2, CT

(g/L NaCI) (WJL NaCI)

1

2

345678910

1112131415

1617181920

04-05-11

05-03-1106-07-1107-12-1108-09-1109-13-11

10-04-1110-11-1111-08-1112-06-1101-10-1202-07-1203-13-1204-10-12

04-10-1205-08-1205-08-1206-05-12

07-10-1208-07-12

1.07

1.071.051.061.061.061.07

1.061.051.061.081.08LOS1.07

1.071.061.051.051.07

1.08

1.07

1.07

1.071.07L071.071.071.071.07

1.061.071.071.071.06

1.061.061.061.061.061.06

State and USEPA

S Control Limits SAM

CT-2S CT+2S

0.01 1.05 1.09 0.090.01 1.05 1.09 0.09

0.01 1.05 1.09 0.090.01 1.04 1.09 0.090.01 1.04 1.09 0.090.01 1.04 1.09 0.090.01 1.04 1.09 0.090.01 1.05 1.08 0.090.01 1.05 1.08 0.09

0.01 1.04 1.08 0.090.01 1.04 1.09 0.090.01 1.04 1.09 0.090.01 1.04 1.09 0.09

0.01 1.05 1.08 0.090.01 1.05 1.08 0.090.01 1.05 1.06 0.090.01 1.05 1.08 0.090.01 1.05 1.08 0.090.0.1 1.05 1.08 0.090.01 105 1.08 0.09

LaboratoryWarning Umits

CT - S&. CT * SX0

0.980.98

0.980.980.980.980.980.98

0.980.980.980.980.980.98

0.980-980.980.980.98

0.98

1.161.161.161.15

1.151.151.151.151.151.15

1.151.151.15

1.15

1.15

1.151.15

1.151.15

215

0.180.180.180.180.18

0.180.180.180.180.180.180.180.18

0.180.180.180.18

0.180.180.18

LaboratorySA25 Control Limits

CT- SAý CT + Sz

USEPA

SAL3 Warning LimitsCT- SA-s CT+ SA.7

USEPA

S,.O Control LimitsCT - S,, CT+Sjue

CV

0.890.890.89

0.890.890.89

0.890.880.880.880.880.880.88

0.880.880.880.880.88

0.880.88

1.251.251.25

1.251.25

1.251.251.251.25

1.251.251.251.251.25

1.241.241.241.241.24

1.25

0.480.480.48

0.480.48

0.480.480.480.480.48

0.48O.480.480.480.48

0.480.480A80A80.48

0.590.590.59

0.590.590.590.590.590.590.59O.59

0.590.59

0.590.590.590.58

0.580.590.59

1.551.551.551.551.55

1.551.55

1.551.541.541.541.551.551.54

1.54

1.541.541.541.54

1.54

0.660.66

0.660.660.660.660.66

0.660.660.660.660.660.660.66

0.660.660.660.660.66

0.66

0.410.41OA10.410.410.410.410.400.40

0.400.400.400.400.400.40

0.400.400.400.400.40

1.731.731.73

1.731.73

1.731.731.731.731.72

1.731.731.731.721.721.72

1.721.721.72

1.72

0.010.010.010.010.01

0.010.01

0.010.010.010.010.010.01

0.010.010.010.01

0.010.010.01

Note: 7-d c1, = 7-day 25% inhibition concentration. An estimation of the concentration of sodium chloride that would cause a 25% reduction in Ceriodaphnlo reproduction for the test population.CT = Central tendency (mean IC-s).

S = Standard deviation of the ICn values.

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

Sý. = Standard deviation corresponding to the 10e percentile CV. (SAe. = 0.08)

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

IJSEPA Control and Warning Umits

SAJ. = Standard deviation corresponding to the 75h percentile CV. (S&75 = 0.45)

SeA"= Standard deviation corresponding to the 9 0e percentile CV. (S, = 0.62)

CV = Coefficient of variation of the IC, values.

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

File: CdNca0CCRO8Ol72icdsxEntered by: J. Sumnjr

Reviwemd by-.:

ET:Environmental Testing Solutions, Inc.


Precision of Endpoint Measurements

40

00

0I.'

0

00

*U

M

35

30

25

20

15

I I I I I I I J I I I I I I I I I i I I

Minimum Acceptance Criteria (> 15.0 offspring per surviving female)

I I I I I I I I I I I I I I I I I I

•o40

.0 O 30

o-20

~10

50

North Carolina Acceptance Limit (< 40.0%)

Kentucky Acceptance Limit (< 30.0%)

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

40

30

20

10

11

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

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

S................... .......... ........ I I

V Ns 'N' ,'SAI, 'bx'ý I I.O'vs sqýv , 'NSN NN -N'* 1'ý- NAN N-N"' %A-" 0'* OINI - -o1b bg-" "b"i "'b" ANý% NTI 'bT lo, lo, 'o-1 Q'S"Test date

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

- -- Central Tendency (mean Control Reproduction, CV, or PMSD)......... Control Limits (mean Control Reproduction, CV, or PMSD ± 2 Standard Deviations)I . 4- ý

f -nlet .

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

Reviewed by: -

" lPrecision of Endpoint Measurements

.E T S 1Ceriodaphnia dubia

,: IChronic Reference Toxicant Data


Test Control Control MeanTest date CT CV CF MSD PMSD (.Tinumber Survival Reproduction

for Control Mean for Control(N) (offspring/female) Reproduction (K) Reproduction (%) for PrVlSD (%)

(offspring/female) CV (%)

1 04-05-11 100 33.0 5.9 2.2 6.72 05-03-11 100 30.9 32.0 4.5 5.2 1.9 6.1 6,43 06-07-11 100 28.9 30.9 6.7 5.7 2.5 8.7 7.24 07-12-11 100 32.2 31.3 6.0 5.8 2.0 6.2 6.9

5 08-09-11 100 28.3 30.7 5.0 5.6 1.9 6.8 6.9.6 09-13-11 100 32.1 30.9 8.2 6.1 1.9 5.8 6.77 10-04-11 100 32.8 31.2 6.1 6.1 2.1 6.5 6.78 10-11-11 100 31.3 31.2 3.0 5.7 2.1 6.6 6.79 11-08-11 100 29.0 30.9 7.6 5.9 1.8 6.1 6.6

10 12-06-11 100 29.3 30.8 5.8 5.9 1.9 6.5 6.611 01-10-12 100 32.4 30.9 5.1 5.8 2.4 7.4 6.7

12 02-07-12 100 31.7 31.0 4.2 5.7 2.1 6.5 6.713 03-13-12 100 31.1 31.0 7.2 5.8 1.9 6.1 6.6514 04-10-12 100 29.4 30.9 6.8 5.9 2.4 8.0 6.715 04-10-12 100 30.3 30.8 7.1 6.0 2.7 8.8 6.13

16 05-08-12 100 31.7 30.9 6.1 6.0 1.9 6.1 6.1317 05-08-12 100 32.4 31.0 5.1 5.9 2.0 6.1 6.318 06-05-12 100 32.1 31.1 4.0 5.8 2.3 7.1 6.819 07-10-12 100 31.2 31.1 4.7 5.8 2.0 6.3 6.18

20 08-07-12 100 29.2 31.0 3.5 5.6 1.9 6.6 6.7

Note: CV = Coefficient of variation for control reproduction.

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

Upper CV bound determined by USEPA (90'h percentile) = 42%


PMSD is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment thatcan be declared statistically significant In a whole effluent toxicity test.Lower PMSD bound determined by USEPA (10th percentile) = 13%.Upper PMSD bound determined by USEPA (90th percentile) = 47%.

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

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

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

File: CdNaCICRO080712.xlsx

Entered by: J. Smnner

(t age 89 of 99 Reviewed by: _*-

0ETS

Page I of 6

/'i

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

Species: Ceriodaphnia dubia

CdNaCICR #: I.•

Dilution preparation in ormation: Comments:NaCI Stock INSS number. ltyA. __n_

Stock preparation: 100 g NaCI/L:Dissolve 50 g NaCI in 500 mL Milli-Q water.

Dilution prep (mg/L) 600 800 1000 1200 1400Stock volume (mL) 9 12 15 18 21Diluent volume (mL) 1491 1488 1485 1482 1479Total volume (mL) 1500 1500 1500 1500 1500

Test organism source information: Test information:Organism age: < 24-hours old Randomizing template color .,•Date and times organisms were bom mGL.01t U. 01% M toW Incubator number and shelfbetween: location:Culture board: -

Replicate number: 2 3 )4 15 16 17 a 9 10 YWT batch:Culture board cup number- • I 'I 1 oi 6% 110 1- 010_________

Transfer vessel infbrmation: pH= i".61 S.U1 Temperature = 2q.t *C Selenastrum batch:Average transfer volume (mL): 0.0 51 -A

Daily renewal information:

Day Date Test initiation and feeding, MHSW Analystrenewal and feeding, or batch used

termination time

0 ot.-.(t. tOi- oI -OsrL _

2 0.f_____-a

.___5 oiq1S 04-0-h11-~ 4 ..6 . .O - . 1"I. CA •.\ , t 0

Control information: Acceptance criteria Summary of test endpoints:% of Male Adults: 0 .20% 7-day LC50 51400% Adults having 3 rd Broods: INA 80% NOEC 000% Mortality: . f7. :520% LOEC I• t3Mean OffspringFemale: 21 . > 15.0 offspring/female ChV 101K.

%CV: ___ _,S7.1 <40.0 % IC 25 161

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

S6ETS

Page 2 of 6

Species: Ceriodaphnia dubiaCONTROL /

CdNaCICR #: VbtSurvival and Reproduction Data

II

- I .... Replicate numberDay 1n 11 2 3 4 5 6 7 8 9 10

21 Young produced I....bi..l . 1l o o I-3- 600 1.~.Adult mortality I %. I t- L - L- i -

Youngprodueed n in II n 0-

___ Adult mortality [__ *.-'%-I L- ~II~ t-- I %- 'I % -- I '-. FL I--3 Young produced

Adult mortalitg •.•q a

I Adult mortality

Tot5l young produced "T..."1

Final Adult Mortality CT f ' i._.._. •C_ - . . '-

X for ung roods __ , C > -

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

A % Mortality:

Mean Offs rin eFmale: [ ",1.Lt-600 NaCI/L Survival and Reproduction Data

I ~" Replicate number ____ ___

4 %S I'S

Day' 1d 2 3 4 - 6 [ 8 9 10

Toa Iyoung produced 3 .v% 1w ft __IL1 __ -LI

Adult mortality 17 _N _7

2 Y oungproduced jJ ol lo )o .. •I() Q. I oN Adult mortality I .LIt b- o (l.s I brodI s.p.li b--1 t1i

Young producedl 1 10 0 t r Ins0 I r

I_ Admlt mortality LFWT•. JC]-- • t I [T7I I '- I- 1 I,4 Young produced i~ S 113"1 I•"I• 1•I •" I '.I Adult mortality '-. 1 ', I I 'I% Mot ality [107.

Adultmortallty 'I__l. JMe 1.f. iJ. n 1±1 [

60 mVug NC/ uvvladRproducto Data:• Iq • • , • • 'Day 1 J 1__ 314 1 8_ 9 10

It Young produced j oi . 0 -- 3 0 "5 1 20

l__ Adult mortality IA~ L I L- I ['Lr L L L.._ L i- L2 Young produced (n 4 *-0II -

Toa Young produced 1 .0~3 ~~ b 2 .

Final Adult Mortality . . . TJ - t. ,.... ... t.Note: Adult mortality (L = live, D L dead), SB = split brood (single brod split between two days), CO = carry over (offspringcarried over with a dluring ansfer).

Concentration:% Mortality: 07.Mean Offspring/Female: 0o.2-.% Reduction from Control: -.a 7.

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

%p ge 91 of 99

6ETS

Page 3 of 6

Species: Ceriodaphnia dubia800 ma NaCI/L

CdNaCICR #: I_&Survival and Reproduction Data

Replicate numberDa y 1 2 3 4 5 6 7 8 9 10

I Young produced 0 0.. 1) 0 0C1 0_ Adlmortality[ ~ N.- _ I~

2 Young producedAdult mortality _ __ . • a . .. . ',. s..

3 oung produced 0 ) - 0J O0 01 0Adult mortality __, - I I I ___________________________4 Young produced "%I t,

Adult mortalityS Young produced 103J ~I~Ii 17-1 06I__ 2. C

Adult mortality %._ " _ .... •q-.6 oung produced __ __S_ _

Adult mortality _-_- • _ • I I7 Young produced -

Total young produced . z "%..w B0% 'Lt

Final Adult Mortality X__ _.. . -%._ j_ _ .Note: Adult mortality (L = live, D = dead), SB split brood (single brood split between two days), CO = earry over (offspringcarried over with adult during transfer).

Concentration:% Mortality: 07.Mean Offspring/Female: .30A% Reduction from Control: k 17. -

1000 mg NaCI/L Survival and Reproduction DataReplicate number

Day 1 2 3 4 5 6 7 8 9 1 101 Young produced ] , - I- - - i - -

Adult mortality t,.-*_ - " _ _ I '

2oa Young produced o• n} b 6 ••"- '" • •• "

nalAdult mortality.L L I U-.- UI. L,-3f Young produced C) I e) Cp

oe Adult mortality W t L Iro I LrIoL spItw w rr ia4 Young produced • JA s6 I I

Adult mortality JL %-~1 .~J .J... - I5 Young prdue 0C 1_ '1,t 10 IP I1.

[Adult mortality V_ _ .- . _ _ _ _ _ _

.6 Young produced ti 0. 0 0 0 ' __

Adult mortality X4_ I__ LJ__%_I '- I V-- I -I %-7 Young producedJ %S 'S ~ IS 16 17'. IS, '- i-

Final Adult Mortality 7-k L -'Note: Adult fliortality (L = live, D =dead), SB =split brood (single brood split between two days), CO =carry over (offspringcarried over with adult during transfer).

Concentration:% Mortality: 10.qMean Offspring/Female: 24.q

* age 92 of 99

I% Reduction from Control: ' .'ISOP AT14 - Exhibit AT14.1, revision 06-01-11

0ETS

Page 4 of 6

Species: Ceriodaphnia dubia1200 me NaCI/L

CdNaCICR #: _

Survival and Reproduction Data

Replicate number

1 Young produced Q Qt IIIAdult mortality 1 .3 4 5 6 7 8 9 10

1 Young produced 1Adult mortality ___ A.--'-__ - -P I' 1_-'-3 Yugpoue0- n 0_l

_J Adult mortality I~I _it C4CJ4 Young produced I.•jZ• 1 2I ,o I .- L j I I__IAdult mortality U J J' I'- ' __ L ._

5 oung produced I I I_ Adult mortality. I 1'".- ['- "FTIL-" -.-..

Adult mortality __,_.. • - '- _ .. _ ___- _

7 Young produced _ 5 i --I -1 %ATotal young produced jA I1S 12- 66 1 16

Final Adult Mortality -Q- %? N %-- -t %.- %.- % ..-Note: Adult mortality (L = live, D = dead), SB = split brood (single brood split between two days), CO = carry over (offspringcarried over with adult during transfer).

Concentration:% Mortality: 0?.

•.Mean Offspring/Female: %.L% Reduction from Control: SI'&.

1400 mg NaCI/L Survival and Reproduction DataReplicate number

Day 1 1 2 3 4 5 6 7 8 9 101I Young produced

Adult mortality I . .. L QL CL C 1 E. t2 Yongproduced cc) I rr () iaSAdult mortaliayt y• _I--I• I I I L _3 Young producedI 0 . • _ O I I

'4 Young produced - • . .

Adult mortality

3 Young producedAdult mortality I IU I. LI_ u,._1 L. L. ... i

6 Young produced 0J.JA I~ IL t ~ 1.. 0.~__ Adult mortalityj l .4U L IL L U j -

7 Young produced _,,, _ . X 1 __-_

Total young produced Lk A 10S01 L j C) .

Final Adult Mortality _ • • • . . .Note: Adult mortality (L = live, D =dead), SB = split brood (single brood split between two days), CO = carry over (offspring

age 93 of 99

Concentration:Mortality: 07.

Meann Offilspring/Female: 1,,.0

I% Reduction from Control: 71. $1.4SOP AT14 - Exhibit AT14. 1, revision 06-01-11

!E TSF nvlnrnmental Testing Solutions, Inc.

Verification of Ceriodaphnia Reproduction Totals

Control

Day Replicate number - TotalDy 1 21 3 4 5 6 7 8 9 101 0 0 0 0 0 0 0 0 0 0 0

2 0 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 0

4 4 5 4 4 5 5 6 4 5 5 475 11 0 10 10 10 10 12 10 10 10 936 0 10 0 0 0 0 0 0 0 0 107 15 13 15 15 14 15 13 16 13 13 142

Total 30 28 29 29 29 30 31 30 28 28 292

600 mg NaCI/L

Day Replicate number - - Totalay 1 2 3 -4 5 ,6 7 8 9 1 oa1 0 0 0 0 0 0 0 0 0 0 0

2 0 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 04 5 5 4 6 6 4 6 5 5 5 515 11 0 10 10 13 11 11 10 13 10 996 0 11 0 0 0 0 0 0 0 0 117 14 13 14 14 1 1 4 13 13 141

Total 30 29 28 30 33 31 33 29 31 28 302

800 iNg aCi/L

Day Replicate number Total1 2 3 4 5 6 7 8 9 101 0 0 0 0 0 0 0 0 0 0 0

2 0 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 0

4 5 5 4 4 4 5 4 6 6 5 485 11 11 10 11 10 12 10 13 12 10 1106 0 0 0 0 0 0 0 0 00 07 14 14 14 13 13 16 15 17 13 14 143.

- 301Total 30 30 28 28 27 33 29 36 31 29 301

1000 mg NaCI/LReplicate number TotalDay 1 2. 3 4 6 7 8 19 1 T10

1 0 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 0

4 5 4 6 4 4 4 4 3 4 4 425 0 12 11 11 10 10 10 10 10 10 946 11 0 0 0 0 0 0 0 0 0 117 15 15 13 15 12 14 13 13 13 14 137

Total 31 31 30 30 26 28 27 26 27 28 284

1200mg NaCLa

Day Re cate number TotalI _ 1 2 3 4 5 6 7 8 9 10

1 0 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 10 .0 0 0 0

3 0 0 0 0 0 0 0 0 0 0 04 3 1 2 2 3 2 1 3 3 3 235 7 0 6 4 5 8 5 6 9 5 556 0 9 0 0 0 0 0 0 0 0 97 4 6 5 3 5 2 7 4 4 5 45

Total 14 16 13 9 13 12 13 13 16 13 132

1400mni NaCIILRe plicate number Total

Day 1 2 3 4 5 6 7 8 9 10

1 0 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 0 04 1 1 1 2 2 3 2 3 3 1 195 0 0 2 3 0 2 4 1 2 2 166 1 3 0 0 2 0 0 0 0 0 67 2 2 1 1 1 3 2 3 1 3 19

Total 4 6 4 6 5 8 8 7 6 6 60

File: CdNaCICR_080712.xlsxEntered by: J. SumnerReviewed by:

CA0

too Ceriodaphnia dubia Chronic Reference Toxicant Test* EPA-821-R-02-013, Method 1002.0

is Environmental Testing Solutions, Inc. Quality ControlVerification of Data Entry, Calculations, and Statistical Analyses

Test number:

Test dates:

CdNaCICR #138

August 07-14,2012

Concentration Replicate number Survival Average reproduction Coeffident of Percent reduction from(mg/L NaO) 1 2 3 4 5 6 7 8 9 10 (%) (offspringfemale) varIatton (%) control (%)

Control 30 28 29 29 29 30 31 30 28 28 100 29.2 3.5 Not applicable

600 30 29 28 30 33 31 33 29 31 28 100 30.2 6.0 -3.4

800 30 30 28 28 27 33 29 36 31 29 100 30.1 8.9 -3.1

1000 31 31 30 30 26 28 27 26 27 28 100 28.4 6.9 2.7

1200 14 16 13 9 13 12 13 13 16 13 100 13.2 15.1 54.8

1400 4 6 4 6 5 8 8 7 6 6 100 6.0 23.6 79.5

Dunnett's MSD value:PMSD:

1.9296.6


PMSD is a measure of test precision. The PMSD is the minimum percent difference between the control and treatment that can be declared statisticallysignificant in a whole effluent toxicity test.

Lower PMSD bound determined by USEPA (I0 th percentile) = 13%.

Upper PMSD bound determined by USEPA (90'h percentile) = 47%.Lower and upper PMSD bounds were determined from the 10th and 90th percentile, respectively, of PMSD data from EPA's WET InterlaboratoryVariability Study (USEPA, 200 1a; USEPA, 2001 b).

USEPA. 2001a, 200lb. Final Report: Interlaboratory Variability Study of EPA Short-term Chronic and Acute Whole Effluent Toxicity Test Methods, Volumes I and 2-Appendix EPA-821-B-01-004 and EPA-821-B-01-005.US Environmental Protection Agency, Cincinnati, OH.

File: CdNaCICR_080712.xlsxTable populated from associated 'Verification of Cedodaphnia Reproduction Totals" spreadsheet.

Spreadsheet entered by: J. SumnerReviewed by: 1

SETS6 nWrwrmWeazft oou~wwIm


Ceriodaphnla Survival and Reproduction Test-ReproductionStart Date: 8/712012 Test ID: CdNaCICR Sample ID: REF-Ref ToxdcantEnd Date: 8/14/2012 Lab ID: ETS-Envlr. Testing Sol. Sample Type: NACL-Sodium chlorideSample Date: Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Cariodaphnia dublaComments:Conc-mgIL 1 2 3 4 .6 6 7 8 9 10

D-Control 30.000 28.000 29.000 29.000 29.000 30.000 31.000 30.000 28.000 28.000600 30.000 29.000 28.000 30.000 33.000 31.000 33.000 29.000 31.000 28.000800 30.000 30.000 28.000 28.000 27.000 33.000 29.000 36.000 31.000 29.000

1000 31.000 31.000 30.000 30.000 26.000 28.000 27.000 26.000 27.000 28.0001200 14.000 16.000 13.000 9.000 13.000 12.000 13.000 13.000 16.000 13.0001400 4.000 6.000 4.000 6.000 5.000 8.000 8.000 7.000 6.000 6.000

Transform: UntransformedConc.mg/L Mean N-Mean Mean Min Max CV% N

D-Control 29.200 1.0000 29.200 28.000 31.000 3.537 10600 80.200 1.0342 30.200 28,000 33.000 6.005 10800 30.100 1.0308 30.100 27.000 36.000 8.921 10

1000 28.400 0.9726 28.400 26.000 31.000 6.884 10'1200 13.200 0.4521 13.200 9.000 16.000 15.067 10'1400 6.000 0.2055 6.000 4.000 8.000 23.570 10

Rank 1-TailedSum Critical

121.00 75.00111.50 75.0092.00 75.0055.00 75.0055.00 75.00

IsotonicMean N-Mean29.833 1.000029.833 1.000029.833 1.000028.400 0.952013.200 0.44256.000 0.2011

Auxiliary Tests Statistic Critical Skew KurtKolmogorov D Test indicates non-normal distribution (p <= 0.01) 1.17686 1.035 0.53863 0.89662Bartlett's Test indicates equal variances (p = 0.14) 8.40619 15.0863Hypothesis Test (1-tail. 0.05) NOEC LOEC ChV TUSteers Many-One Rank Test 1000 1200 1095.45Treatments vs D-Control

Linear Interpolation (200 Resamples)Point ma/L SD 95% CL SkewIC05 1000.77 44.7051 868.009 1015.87 -1.6465IC10 1020.39 9.61319 991.422 1034.38 -1.3975IC15 1040.02 7.68096 1020.26 1053.25 -0.3510 1.o-IC20 1059.65 7.19436 1041.68 1071.64 -0.3336 0.9IC25 1079.28 6.83735 1063.08 1090.45 -0.3157IC40 1138.16 6.71529 1123.99 1148.74 -0.2616 0.8

IC50 1177.41 7.42545 1162.25 1189.63 -0.2048 0.7

0.6

0.4

0.3

0.2

0.1.

0.0 ------

-0.1 10 500 1000 1500

Dose mg/L

Dose-Response Plot

30

25

20

10

5

(fage 96 of 99

File: CdNaCICR0807I2.xlsxEntered by: J. !;unmerReviewed by. _-_

"ETS(a Environmentl Tesoti ons, Inc.


Ceriodaphnia Survival and Reproduction Test-ReproductionStart Date: 8/7/2012 Test ID: CdNaCICR Sample ID: REF-Ref ToxicantEnd Date: 8/14/2012 Lab ID: ETS-Envir. Testing Sol. Sample Type: NACL-Sodium chlorideSample Date: Protocol: FWCHR-EPA-821-R-02-013 Test Species: CD-Ceriodaphnia dubiaComments: Used for PMSD calculation only.Conc-maiL 1 2 3 4 6 6 7 8 9 10

D-Control 30.000 28.000 29,000 29.000 29.000 30.000 31.000 30.000 28,000 28.000600 30.000 29.000 28.000 30.000 33.000 31.000 33.000 29.000 31.000 28.000800 30.000 30.000 28.000 28.000 27.000 33.000 29.000 36.000 31.000 29.000

1000 31.000 31.000 30.000 30.000 26.000 28.000 27.000 26.000 27.000 28.0001200 14.000 16.000 13.000 9.000 13.000 12.000 13.000 13.000 16.000 13.0001400 4.000 6.000 4.000 6.000 5.000 8.000 8.000 7.000 6.000 6.000

Transform: Untransformed 1-TailedConc-mg/L Mean N-Mean Mean Min Max CV% N t-Stat Critical MSD

D-Control 29.200 1.0000 29.200 28.000 31.000 3.537 10600 30.200 1.0342 30.200 28.000 33.000 6.005 10 -1.186 2.287 1.929800 30.100 1.0308 30.100 27.000 36.000 8.921 10 -1.067 2.287 1.929

1000 28.400 0.9726 28.400 26.000 31.000 6.884 10 0.948 2.287 1.929*1200 13.200 0.4521 13.200 9.000 16.000 15.067 10 18.969 2.287 1.929*1400 6.000 0.2055 6.000 4.000 8.000 23.570 10 27.505 2.287 1.929

AuxillaryTests Statistic Critical Skew KurtKolmogorov D Test indicates non-normal distribution (p <= 0.01) 1.17686 1.035 0.53863 0.89662Bartletts Test indicates equal variances (p = 0.14) 8.40619 15.0863Hypothesis Test (1 -tail, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob CIfDunnett's Test 1000 1200 1095.45 1.92879 0.06605 1109.51 3.55741 1.6E-38 5,54Treatments vs D-Control

*age 97 of 99

File: CdNaCICR_0807 [2.xlsxEntered by: J. SumnerReviewed by: _A-

.. I.,. . ... .:.e.5 ,.:6 Page 5 of 6

Species: Ceriodaphnia dubia

Daily Chemistrv:

CdNaCICR #: IVb

• • "Day(Aayst identified for each dayr performed pHl, D.O. and conductivity measure'ments only.

Concentration ParameterPH (s.u.) [

Conductivity(gmihos/cm) 0

CONTROL *Alkalinity(Ing CaCO3/L)*Hardness(mg CaCO3 L)"* Temperature (*C). ' " .•A• . 1¢4. 'i -. 1pH (S.U.) 1.7C .23 I _"

DO (mg/L) 7.7 9" D600 mg NaCIIL Conductivity

(tuhhos/cm) 14t__'_13__ _ _ *Temperature(*C) -Lq A U_0 , . . C)

pH (S.U.) M.U 147DO (mgIL) 7.1

800 mg NaCI/L Conductivity

* Temperature (9C) 1I4.0, . 0 1. 0 all

pH (S.'U.) -7.77____

1000 mg NaCI/L Conductivity

*Temperature (IC) IS. .0 ,SO - "S._,_ "-s. -pH (S.ff.) "7,77/ f .•4 I••Z

' •. 7.•7.S 0 8._1200 mg NaCI/L Conductivity

I *Temperature (OC) 75,.0 . ..- .1., I S. .

pH (S.U.) 7.90 "4.91- - . * ~ I -DO (mg/L)

1400 mg NaCIIL Conductivity ' o(g±mhos/c) ________

*Temperature (SC)

STOCK Conductivity t_ _ 1~ -gno/m

Initial I1 1

*Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily RenewalInformation table located on Page 1. Alkalinity and hardness performed by the analyst identified on the bench sheet specific for eachanalysis and transcribed to this bench sheet by:

SOP AT14 - Exhibit AT14.1, revision 06-01-11-em.-,b~,wby

Kt.,EKnm

e 98 of 99

S

S0ETSE~maIT~OgSOI**b~

Page 6 of 6

Species: Ceriodaphnia dubia CdNaCICR #: I b t"

Day(Analyst identified for each day, performed PK- D.O. and conductivity measurements on l,.

3 4 5 64Analyst zPl orBMjoa

Concentration ParameterpH (S.u.) "-CS 7.S 1.S _. _ _ . . S .5 7.i . *'t.

DO (mg•L) 77 -.0 7..Conductivity(tunhos/cm)3o31*Alkalinity

CONTROL *Aklinity(Ing CaCO3/L) t 2*Hardness(Ing CaC03/L)

*Temperature " 0.0 " -o A 'L L Vi.A .0.*\ .Lblu aAA

DO (mg/L) 7.q

S600 mg NaCIIL Conductivity I~)___ __

*Temperature ..• t j '*z .. 8,

pH (S.U.) -" .75 -7.'06 "___ 196 .2.1 . 2.±•I -z.DO(mg/L) , .o -.q 2 ._

800mg NaCI/L Conductivityo ,o mgmhos/cm _

* Temperature "q• "$ S LO "t,• •7:t "•" L,I) MY _T_

pH (S.U.) /1 -1P I .7.-77 I 7.93 '7.2 1 -7 It 1~ 1. -1. 1 A.n 7.7"L- U *-'E~-4-'---- II - - 4 __________ - 4 4I-.L~~'---4 M.Z .11DO (ma/L) W-4 11

1000 mg NaCI/L Conductivity r(jImhostcm) 1 __6 _0

i 03O

"U.0 u1.5*Temperature(00) -0.1 11 -LS. 1) 11%S.% IpH (S.U.) ," "7.2I ".Lq 7.7' 811." .. ".Z 7.7DO (rag/L) • . . . 7= , )-• 7.

1200 mng NaCL/L Conductivity(ginhos/cmn) 9141*Temperature " "

.. PH (S.U.) 13-- -7• ". $ /q 7.q1"j { '• • . '

DO (mg/L). , 7.q 8.o1400 mg NaCIIL Conductivity

*Temperature "•• .,L . Lq.• '.• -Let -Lq. "T.. 1 1, ¢CIL

Initial Final I- Initial 11 Final I Initial 11 Final I Initial 11 Final I- U a - r U m .0

*Temperatures performed at the time of test initiation, renewal or termination by the analyst identified in the Daily RenewalInformation table located on Page 1. Alkalinity and hardness performed by the analyst identified on the bench sheet specific for eachanalysis and transcribed to this bench sheet by:

*Z. ge 99 of 99SOP ATI4 - Exhibit ATI4.1, revision 06-01-11

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