Tennessee Valley Authority, Post Office Box 2000, Soddy Daisy, Tennessee 37384-2000 October 10, 2012 Ms. Christina Morgan Tennessee Department of Environment and Conservation Division of Water Pollution Control Enforcement & Compliance Section 6 th Floor, L & C Annex 401 Church Street Nashville, Tennessee 37219 Dear Ms. Morgan: TENNESSEE VALLEY AUTHORITY (TVA) - SEQUOYAH NUCLEAR PLANT (SQN) - NPDES PERMIT NO. TN0026450 - DISCHARGE MONITORING REPORT (DMR) FOR SEPTEMBER 20"12 AND REVISED DMR FOR AUGUST 2012 Enclosed is the September 2012 Discharge Monitoring Report for Sequoyah Nuclear Plant. The toxicity report and revision to Discharge Number 101T, Biomonitoring for Outfall 101 for August 2012, are also attached. There were no exceedances during the monitoring period. If you have any questions or need additional information, please contact Brad Love by email at [email protected]or by phone at (423) 843-6714. I certify under penalty of law that this document and all attachments were prepared under my direction or supervision in accordance with a system designed to assure that qualified personnel properly gather and evaluate the information submitted. Based on my inquiry of the person or persons who manage the system, or those persons directly responsible for gathering the information, the information submitted is, to the best of my knowledge and belief, true, accurate, and complete. I am aware that there are significant penalties for submitting false information, including the possibility of fine and imprisonment for knowing violations. Sequoyah Nuclear Plant Enclosures cc (Enclosures): Chattanooga Environmental Field Office U.S. Nuclear Regulatory Commission Division of Water Pollution Control Attn: Document Control Desk State Office Building, Suite 550 Washington, DC 20555 540 McCallie Avenue Chattanooga, Tennessee 37402-2013
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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
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
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
COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)
Toxicity was not sampled in September 2012.
EPA Forim 3320-1 (REV 3199) Previous editions may be used Page 1 of 1
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
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/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
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)
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
COMMENTS AND EXPLANATION OF ANY VIOLATIONS (Reference all attachments here)
Toxicity sampling began on August 12 and ended on August 17. This is the revised August 2012 DMR data set for 101T. The toxicity report is attached.
EPA Form 3320-1 (REV 3199) Pmvious editions maybe used Page 1 of I
TENNESSEE VALLEY AUTHORITYTOXICITY TEST REPORT
INTRODUCTION / EXECUTIVE SUMMARYReport Date: September 17, 2012
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
*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)
*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
2. Ceriodaphnia dubia
None
DEVIATIONS / MODIFICATIONS TO PRETEST CULTURE OR HOLDING OF TESTORGANISMS
1. Pimephales promelas
None
2. Ceriodaphnia dubia
None
Page 12 of 99
PHYSICAL AND CHEMICAL METHODS
1. Reagents, Titrants, Buffers, etc.: All chemicals were certified products used before 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).
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
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
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
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
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
Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations ofChemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test
Table B-1. Sequoyah Nuclear Plant Diffuser (Outfall 101) Discharge Concentrations ofChemicals Used to Control Microbiologically Induced Corrosion Mollusks, During Toxicity Test
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):
,__ __ ___ __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
SOP G4 - Exhibit G4.2, revision 01-03-12
BIOMONITORING CHAIN OF CUSTODY RECORD Page _1- of -1-
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
SOP G4 - Exhibit G4.2, revision 01-03-12
: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
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.
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.
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.
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.
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
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
"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
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
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
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
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.
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
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
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.
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
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
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
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~
_ 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:
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]
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.
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-
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.
[ 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
C = Larvae weight (mg) = B - A
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
Page 4 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
C = Larvae weight (mg) = B - A
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.
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
*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.
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
Pimephalespromelas Chronic Whole Effluent Toxicity Test
EPA-821-R-02-013, Method 1000.0
Daily Chemical Analyses
Project number: 8207
Concentration Parameter Da 0 Day I Day 2 Da Da y4 Day 5 Day 6_____________________ Initial Final Initial Final Initial Final Initial Final Initial Final Initial. Final Initial Final
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;
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)
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 -
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
Environmental Testing Solutions, Inc.
I ".
Pimephales promelasChronic Reference Toxicant Control Chart
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 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)
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.
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
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
SOP AT21 -Exhibit AT21.1, revision 06-01-11
.ETS(b Envtmnn-r0ITestlng Solutlom In
Page 2 of 5
Species: Pimephales promelas PpKC1CR Test Number: W)$_
Survival and Growth Data
Day Control 450 mg KC[/L 600 m KCI/L-.A B C D. E F G H I J K L
0 101 0 1to It O tO to 0 to I /( O /1 /0
1 10 10 to 1 10to 10 t 10 1.0 £ 0) /0
2 10 tO tO 1O to /o to /O /O to /0 (O
3 (o t ( (t 0 10 to t to 10 10 10 10 (0
4__ _1 10 10 10 10 l. tO 0 qI& 1'A (0 /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
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
Quality ControlVerification of Data Entry, Calculations, and Statistical Analyses
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
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
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
SOP AT21 -Exhibit AT21.1, revision 06-01-11
....... OET
E~wlmmeul'uTesung &olIw~lo.In
Page 5 of 5
Species: Pimephales promelas PpKC1CR Test Number: 21K
Day(Analyst identified for each day, performed pH, D.O. and conductivity measuriements only.)
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)
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.
Ceriodaphnia dubiaChronic Reference Toxicant Control Chart
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)
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
Environmental Testing Solutions, Inc.
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 (%)
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.
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
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.
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
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).
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
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.
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.
*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:
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