Arkansas Analytical, Inc.

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1 Arkansas Analytical, Inc. Toxicity Test Results SHRWOOD NPDS PRMIT NUMBR: AR77 Third Quarter AFIN# -9 Fathead Minnow, Pimephales promelas, Larval Survival and Growth Test Test. Ceriodaphnia dubia, Survival and Reproduction Test Test. Prepared for: Bill Miller Prepared by: Arkansas Analytical, City of Sherwood North Facility 7 -, Bldg, Suite 99. Kiehl AV Little Rock, Arkansas 79 Sherwood AR, 7 Lab Number K9 ATTN: Bill Miller Wednesday, September,

2 Introduction This report contains test results for the toxicity testing for the City of Sherwood. The NPDS permit number is AR77. The plant is located at 9 Algonkin approximately feet east of Indianhead Lakes states subdivision, in Sector, Township North, Range West in Pulaski County, Arkansas. The discharge is to receiving waters named an unnamed tributary of Kellogg Creek, thence to Kellogg Creek, thence to Bayou MTO, thence to the Arkansas River in Segment B of the Arkansas River Basin. The permit requires chronic biomonitoring testing once per quarter for both Ceriodaphnia dubia and Pimephales promelas. The test results in this report represent the testing for the third quarter of. Plant Operations To be provided by permittee. Toxicity testing, Ceriodaphnia dubia and Pimephales promelas, City of Sherwood, Lab# K9, Page

3 Source of ffluent and Dilution Water ffluent samples were collected as follows: Sample Collection: Date, Time Started Date, Time nded Sample #: Sample #: Sample #: 9--,8 9-7-,8 9-8-,8 9-7-,8 9-8-,8 9-9-,8 The samples were composites collected at the final discharge from the City of Sherwood wastewater treatment plant. The following information was collected upon immediate receipt of the samples at the laboratory: Sample Receiving Information: Sample #: Sample #: Sample #; Date, Time Sample(s) Received 9-7-, 9-8-, 9-9-,9 Storage Temperature ( C) Chain of custody documentation is located in Appendix A. The permit designates the receiving water to be used as dilution water for the toxicity tests. Synthetic dilution water was substituted either because zero flow conditions existed or due to an earlier characterization of the receiving water as being toxic. The dilution water used in the toxicity tests was synthetic moderately hard. It was prepared using lga Maxima ultra pure water according to PA specifications. ach batch was analyzed forph, hardness, total alkalinity, and conductivity. Results are provided in Appendix B. Dilution Series Five dilutions in addition to a control (% effluent) were used in the toxicity tests. The dilutions, which were made with synthetic water, were %, %, %, 7%, and %. The low-flow effluent concentration (critical dilution) was defined as % effluent. Toxicity testing, Ceriodaphnia dubia and Pimephaies promelas. City of Sherwood, Lab # K. 9, Page

4 Test Methods PA Method., Fathead Minnow, Pimephales promelas, Larval Survival and Growth Test, was used in this bioassay. Larvae are exposed in a static renewal system for seven days and the results are based on the survival and growth (increase in weight) of the larvae. There were no deviations from the reference method. The test chambers were ml plastic cups, and each chamber contained ten organisms in a test solution volume of mis. The test temperature was degrees Centigrade. Raw data and statistics are provided in Appendix C. PA Method., Cladoceran, Ceriodaphnia dubia, Survival and Reproduction Test, was also used. Neonates are exposed in a static renewal system until at least % of the control organisms have produced a third brood. Results are based on the survival and reproduction of the organisms. One neonate was placed in each often replicate chambers using a randomizing template. Test chambers were ml plastic cups filled with ml of test solution. The test temperature was degrees Centigrade. Raw data and statistics are provided in Appendix D. Test Organisms The organisms used in Test. were < hour old Fathead Minnows, Pimephales promelas, which were purchased from Aquatox; a copy of the organism history is provided in Appendix. The organisms used in Test. were < hour old Ceriodaphnia dubia neonates, (all born within the same eight hours), obtained from an in-house culture. An organism history is provided in Appendix. Toxicity testing, Ceriodaphnia dubia and Pimephales promelas. City of Sherwood, Lab # KJ9, Page

5 Quality Assurance Test Acceptability TST ACCPTANC CRITRIA for Cenodaphnia dubia Control Criteria Greater than or equal to 8% survival Average of or more young per surviving female At least % of surviving females should have produced broods The percent coefficient of variation between replicates must be % or less for the young of surviving females Results % 8. % 9.% Pass X X X X Fail TST ACCPTANC CRITRIA for Pimephales promelas Control Criteria Greater than or equal to 8% survival The percent coefficient of variation between replicates must be % or less for survival Minimum of. mg average dry weight of surviving controls The percent coefficient of variation between replicates must be % or less for growth Results 9%.7%.8.% Pass X X X X Fail Reference Toxicant The reference toxicant used was Potassium Chloride prepared in-house. The tests were performed using moderately hard synthetic as dilution water. The results of the reference toxicant were: RFRNC TOXICANT Ceriodaphnia dubia 8/-9/ Pimephales promelas 8/-9/ NOC Survival: LOC Survival: NOC Reproduction: LOC Reproduction: ppm KC ppm KCI ppm KC ppm KC NOC Survival: LOC Survival: NOC Growth: LOC Growth: SOOppm KC ppm KCi ppm KCI ppm KCI Quality Assurance charts arc provided in Appendix F. Toxicity testing, Ceriodaphnia dubia and Pimephales promelas. City of Sherwood, Lab # K9, Page

6 Summary of Results City of Sherwood Ceriodaphnia dtibia NOC/LOC Survival NOC / LOC Reproduction Mean number of neonates (critical dilution) %CV Reproduction (critical dilution) PMSD Reproduction %/NA %/NA 9. 9.%. Pimephales promelas NOC / LOC survival %/NA NOC / LOC growth %/NA %CV survival (critical.% dilution) Mean dry weight (critical. dilution) in milligrams %CV growth (critical 8.% dilution) PMSD Growth. Conclusion Chronic static renewal larval survival and growth test using fathead minnow, Pimephales promelas, (Method.) The critical dilution is % effluent The effluent samples did not exhibit lethal or sublethal effects at the critical dilution, and, as such, the effluent samples passed both portions of the test. Chronic static renewal survival and reproduction test using Ceriodaphnia dubia, (Method.) The critical dilution is % effluent. The effluent samples did not exhibit either lethal or sublethal effects at the critical dilution, and, as such, the effluent samples passed both portions of the test. Biomonitoring Analysts: Kenneth i Toxicity testing, Ceriodaphnia dubia and Pimephales promelas. City of Sherwood, Lab # K9, Page

7 FATHAD MINNOW LARVA GROWTH AND SURVIVAL PIMPHALS PROMLAS PRMITT: City of Sherwood Sample Collection: Sample #: Sample #: Sample #: Date, Time Started 9--, ,8 9-8-,8 Date, Time nded 9-7-,8 9-8-,8 9-9-,8 Test initiated (date, time): 9-8-, Test terminated (date, time): 9--, Dilution water used: Moderately Hard Synthetic ffluent Cone % % % % % 7% % DATA TABL FOR FATHAD MINNOW SURVIVAL A 8 B c 9 9 D hours hours days cv%.7. ffluent Cone % % % % % 7% % DATA TABL FOR GROWTH OF FATHAD MINNOWS Average Dry Weight in milligrams in replicate chambers A B C D Mean Dry Weight CV%. 8. Coefficient of Variation = standard deviation / mean * Toxicity tesling, Ceriodaphnia dubia and Pimephales promelas, City of Sherwood, Lab # K9, Page 7

8 SUMMARY RPORTING FORMS FOR CHRONIC BIOMONITORING FATHAD MINNOW LARVA GROWTH AND SURVIVAL Pimephales promelas. Dunnett's procedure or Steel's Many-One Rank Test as appropriate: Is the mean survival at 7 days significantly different (p=.) than the control survival for a) LOW FLOW OR CRITICAL DILUTION, (%) YS NO X_. Dunnett's Procedure Is the mean dry weight (growth) at 7 days significantly differenl (p=.) than the control's dry weight (growth) for a) LOW FLOW OR CRITICAL DILUTION, (%) YS NO X. If NO was answered to l.a) and.a) enter [] otherwise enter [!]:_. nter percent effluent corresponding to each NOC below and circle the lowest number: a) NOC survival - % effluent b) NOC growth = % effluent. nter percent coefficient of variation =. % Toxicity testing, Ceriodaphnia dubia and Pimephales promelas, City of Sherwood, Lab # K9, Page 8

9 SUMMARY RPORTING FORMS FOR CHRONIC BIOMONITORING Ceriodaphnia dubia SURVIVAL AND RPRODUCTION Permittee: City of Sherwood Sample Collection: Sample #: Sample #: Sample #: Date, Time Started 9--,8 9-7-,8 9-8-,8 Date, Time nded 9-7-,8 9-8-,8 9-9-,8 Test initiated (date, time): 9-8-, Test terminated (date, time): 9--, Dilution water used: Moderately Hard Synthetic Ceriodaphnia dubia SURVIVAL AND RPRODUCTION NUMBR OF YOUNG PRODUCD PR TST TRMINATION Replicate A B C D F G H J Mean Mean/surviving female cv%* PRCNT % % % xo 8 xo % % x 8..9 % X= Dead Adult; M= Male (Not considered in statistics) *Cocfficient of Variation = standard deviation/ mean * ; CV% calculation based on young per surviving female Toxicity testing, Ceriodaphnia dubia and Pimephales promelas, City of Sherwood, Lab# Kl 9, Page 9

10 SUMMARY RPORTING FORMS FOR CHRONIC BIOMONITORING Ceriodaphnia dubia SURVIVAL AND RPRODUCTION Permittee: City of Sherwood PRCNT SURVIVAL PRCNT Time of Reading: HOURS % % % % 7% % 8 HOURS 8 Test termination 8 9 i. Fisher's xact Test: Is the mean survival at test termination significantly different (p=.) than the control survival for: a) LOW FLOW OR CRITICAL DILUTION, (%) YS NO_. Dunnett's Procedure or Steel's Many One Rank Test: Is the mean number of young produced per female significantly different (p=.) than the control's number of young per female for: a) LOW FLOW OR CRITICAL DILUTION, (%): YS NO. If NO was answered to l.a) and.a) enter [] otherwise enter []:. nter percent effluent corresponding to each NOC below and circle the lowest number: a) NOC survival = % effluent b) NOC reproduction = % effluent. nter percent coefficient of variation = 9. % Toxicity testing, Ceriodaphnia dubia and Pimephales promelas, City of Sherwood, Lab # K9, Page

11 APPNDIX A Chain of Custody Forms

12 7 Interstate, Bldg., Ste. Little Rock, AR 79 PHON: -- FAX: --8 CHAIN OF CUSTODY RCORD CLINT INFORMATION McClelland Consulting ngineers W nd St. Little Rock, AR 7 Attn: Bill Miles BILLING Project Description Turnaround Time Preservation Codes: McClelland Consulting ngineers P.O. Box 87 Little Rock, AR 7-87 Chronic Toxicity Chronic Biomonitoring Reporting Information Telephone: Fax: mail: Hour 8 Hour 7 Hour Routine ( Day) Preservative Code: Bottle Type:. Coo, Degrees Centigrade. Thiosulfiid' Tor Dc-cliloriniiliiin. Sulliiric Acit (H,SO<), >H < S. Hydrochloric Acid(HCI). Nitric Acid (UNO,), ih <. Sodium Hydroxide (NnOH), ]>H -- TST PARAMTRS Jlc.[(lc Typo ( mlc P G - GlJiS. P - I'lWi, V = Scplum. A - Amber / f*^~r' *^C>.XVXV---T^^X/^ Sampler(s) Signature Samplers Printed ^* X"fe^C>n c^t,oo e^7 Arkansas Analytical Work Order Number: Field Number SAMPL COLLCTION Date/s Time/s ^-"V^* %*tv^ Grab Comp tx Numbei of Bottles S Sample Matrix Water SAMPL IDNTIFICATION/ DSCRIPTION City of,"a-/"t-^oo-tx X... K ) -. «,. Relinquished bv: (Siqnaturet ee^ Date/Time ^9 "7 "" ^ / O i»r~. Received bv: (Siqnaturel SAMPL CONDITION UPON RCIPT IN LAB. CUSTODY SALS: v/yes No / (im\^flsl^v&vt'v\s. CONTAINRS CORRCT:. COC/LABLS AGR: Yes Yes No No RMARKS / SAMPL COMMNTS COCL W^fM \PMA vj wkie (/vcir <yvv i-f- 7/7// ' / / - ^/. Relinauished bv: (Slanaturel Date/Time Cj 7 7 i^c T 'jfrvtfstw " ^ ^. Received by lab: (Slanature). PRSRVATION CONFIRMD: P l/jl l^da/. RCIVD ON IC: -i^u \ v^m \. TMPRATUR ON RCIPT: 'wa-x ^ <^A t^>^ Yes Yes /*) / / c/ FOR COMPLTION BY LAB ONLY No No

13 7 Interstate, Bldg., Ste. Little Rock, AR 79 PHON: -- FAX: --8 CHAIN OF CUSTODY RCORD CLINT INFORMATION BILLING Project Description Turnaround Time Preservation Codes: McClelland Consulting ngineers W nd St. Little Rock, AR 7 Attn: Bill Miles McClelland Consulting ngineers P.O. Box 87 Little Rock, AR 7-87 Chronic Toxicity Hour. Cool, - Degrees Cenrigrade. Thiosulfule for Dec lilorinalion Chronic Biomonitoring Reporting Information Telephone: Fax: mail: 8 Hour. Sulfunc Acid (II^O,), ph < S. Hydrochloric Acid(HCI) 7 Hour. Nitric Acid (UNO,), ph <. Sodium Hydroxide (NaOH), pi I - U Routine ( Day) TST PARAMTRS Hoi tic T)' >r Co ill! Preservative Code: C- Glass. I' I'lasiu- Bottle Type: P V = Septum. A- Amber Samplers} Signature x- / Samplers Printed Arkansas Analytical Work Order Number: Field Number SAMPL COLLCTION Date/s a-y- to *,-Z"/G Time/s ^'^** Grab Comp (/ Number ol Bottles j Sample Matrix Water SAMPL IDNTIFICATION/ DSCRIPTION city of ^e^r L~O * <# X K. 9 - b. Relinquished bv: (Sianature) 7/ Date/Time.%r. Received by: (Signature) SAMPL CONDITION UPON RCIPT IN LAB ~Ui rf * CUSTODY SALS: V Yes No / (Art /x-ca- v ^ CONTAINRS CORRCT: Yes No. COC/LABLS AGR: Yes No RMARKS / SAMPL COMMNTS. Relinquished bv: (Sianature) *I«S~-. Date/Time?/?//&. Received by lab: (Signature). PRSRVATION CONFIRMD; Yes vjwcfc cnrttv^^. RCIVD ON IC: -=^ s ^ '. TMPRATUR ON RCIPT: T (s FOR COMPLTION BY LAB ONLY No No

14 7 Interstate, Bldg., Ste. Little Rock, AR 79 PHON: -- FAX: --8 CHAIN OF CUSTODY RCORD CLINT INFORMATION McClelland Consulting ngineers W nd St. Little Rock, AR 7 A tin: Bill Miles BILLING Project Description Turnaround Time Preservation Codes: McClelland Consulting ngineers P.O. Box87 Little Rock, AR 7-87 Chronic Toxicity Hour I. Cool, Degrees Ctnli[;ra(le. Tliiojiulfate for Dcchloriniilion Chronic Biomonitoring Reporting Information Telephone: Fax: mail: 8 Hour 7 Hour Routine ( Day) Preservative Code: Bottle Type;. Sulfuric Acid (,,), ph<. Hydrochloric Acid(HQ). Nitric Acid (UNO,), >ll <. Soilium Hydroxide (NaOH), pll > U TST PARAMTRS BoltkTypr Code P Ci ' Class. P " Plnslii; V - Septum. A = Amber? < > Ss /^ /^7 X Sampler(s) Signature /"~ / (_^ f C < ^» t*-*^o a *S Samplers Printed Arkansas Analytical Work Order Number: Field Number SAMPL COLLCTION Date/s Time/s Grab Comp Number of Bottles Sample Malrix SAMPL IDNTIFICATION/ DSCRIPTION el-~7 ~ve> 8^-7* I/ fl Water city of ^$ A. e, * " u/ G o <J X,, Rellnauishwfcflv: (Si a nature) Date/Time. Received bv: (Signature) SAMPL CONDITION UPON RCIPT IN LAB RMARKS / SAMPL COMMNTS 'fc *? ~~ *? " f fm-+t 7, \' CUSTODY SALS:. CONTAINRS CORRCT:. COC/LABLS AGR: ^ Yes Yes Yes No No No. Rellnauished bv: (Slanaturel Date/Time. Received by lab: (Signature). PRSRVATION CONFIRMD: Yes No / ((jj^ lj&vm/&*vbr^' * lo \ty IL C, la i/i,, /. RCIVD ON IC: CMtAr^^M / J, ],. TMPRATUR ON RCIPT: AT& VY^i^ -L- Yes *c FOR COMPLTION BY LAB ONLY No

15 APPNDIX B ffluent and Dilution Water Data

16 CHMICAL DATA SHT FOR CHRONIC TOXICITY TSTING Fathead Minnow Lab #/ Sample ID Test Start (Date/Time) <%/#// > Client /-/t.a'yvj Test nd (Date/Time) ^//y//^) Day of Test notes/ re marks Control w. D.O. (mg/l) INITIAL ph (s.u.) SS. %Z. L INITIAL M n JL &a :emp (C) INITIAL ru Z- * So ALKALINITY (mg/l) HARDNSS (mg/l) ONDUCTIVITY (umhos/cm) HLORIN (mg/l) CONG: P.O. (mg/l) INITIAL ph (s.u) %- INITIAL ZZ temp (C) INITIAL O^ *rp r^> ^^ i CONG: /7 P.O. (mg/l) INITIAL " ph (mg/l) INITIAL T77 temp (C) INITIAL CONG: D.O. (mg/l) INITIAL 7? ph (s.u.; INITIAL -T7--7, / temp (C) INITIAL 7 CONG: D.O. (mg/l) INITIAL? ph (s.u.) temp (C) CONG: P.O. (mg/l) INITIAL INITIAL INITIAL IOO M w ph (s.u.) INITIAL a l^ ± temp (C) INITIAL CONG: % IV jalkalinityfmg/l) K. jhardnss (mg/l) Sl<- {CONDUCTIVITY (umhos/cm) w<?chlor!n(mg/l) August, 7

17 CHMICAL DATA SHT FOR CHRONIC TOXICITY TSTING Lab # / Sample ID fttx&ftn-7. Test Start (Datemme) Client Test nd (Date/Time) Day of Test ontrol D.O. (mg/l) INITIAL ph (s.u.) INITIAL Vt- xi.emp (C) INITIAL yz^ ALKALINITY (mg/l). HARDNSS (mg/l) ONDUCTIVITY (umhos/cm) HLORIN (mg/l) "7-^ CONG: D.O. (mg/l) INITIAL s 77 I/. ph (s.u) INITIAL - ri TjL ^ O. temp (C) INITIAL CONG: D.O. (mg/l) INITIAL? ph (mg/l) INITIAL ^a. ^7_ temp (C) INITIAL CONG: ZZ) P.O. (mg/l) INITIAL :z_. ph (s.u.) INITIAL. temp (C) INITIAL - CONG: D.O. (mg/l) INITIAL ^X * ph (s.u.) INITIAL o. & Z temp (C) INITIAL CONG: D.O. (mg/l) INITIAL -7 ph (s.u.) INITIAL temp (C) INITIAL ^^ ^ fconc: %!AL KALI NITY (mg/l) loz. -HARDNSS (mg/l) {CONDUCTIVITY (umhos/cm) CHLOR!N(mg/L) ncft Cerodaphnia Dubia notes/remarks August, 7

18 APPNDIX C Fathead minnow raw data and statistics

19 SURVIVAL DATA FOR FATHAD MINNOW LARVAL SURVIVAL AND GROWTH TST LAB #/ SAMPL ID fqocftool TST START DAT q /$fld TIM jtfo CLINT ^ TST ND DAT <^7«7/fTIM ///fxr AG AND SOURC OF MINNOWS DAY (NUMBR SURVIVING) CONC: n CONG?/ CONC:Hy ICONCtfn ICONCfTC CONC:CO ANALYST DAT: TIM: RP# A B C D RP# A B C D RP# A B C D RP# A B C D RP# A B C D RP# A B C D start tp * - start D -^ start f*' start p -^ start \b *~ start ^ lo a ** rp hilh fath [ \ In - D [fi aio to \& i&o /D f [^(n a WV) P I i a M> q//. M" /C ( Q g IT? ( IV InQ ' C> $ J& ffi Q IP O \n o [ IQ if)!o O 9 r^ OT IPO -C n 'QD D O lg D r/ (r) /^ 9 a L^ in m ^ Q D < > '/I/ID si Q «(> u If) tt P P Q P,& n i If) i $ ii P s /) m Q (; a a Qttlltf) JflPT U '/' ^i /^A) L /f; [ UH Ifi If) O Kz C o P n._ If) }! ffi *^ to its B In a ID v J (^ /D In ' I Q l P Sa ID Z fl Icf f//v//ft CV = PRCNT COFFICINT OF VARIATION: STANDARD DVIATION/MAN * D LA 9 I ^ 7 / 'o 'Qr) SURVIVAL 7 % ^ 7 & ID P b 7 X b IP 7 H W d 7 /() J //7/ //r^r I(OD ^ /GD err) % ify-o te?& \oo ( *VS OT> % ion Cfo G GTi im % L^o ^( ti> <HQ % i&o DV gs //on % l/vo /(9O W> Ico %& MAN % %> MAN % CV MAN % \b MAN % Tf MAN % ^ MAN % V g^ ^K cv b77/ cv cv cv cv w «jggjjjib^ji ^^^^*^^^9 July 7

20 Pimephales promelas FATHAD MINNOW TST. WIGHT DATA FOR LARVAL SURVIVAL AND GROWTH TST LAB#/#s: K 9 CLINT: City of Sherwood ANALYSTS: KP SAMPL ID: - RP# A B C D DRY WIGHT TIN+LARVA (9) INITIAL WIGHT TIN (g) TOTAL DRY WIGHT OF LARVA (9) TST DATS (BGIN / ND): 9/8-/ WIGHING DAT / TIM: 9//, DRYING TMP (DGRS C): DRYING TIM (HOURS): DRY WIGHT NUMBR OF LARVA OF LARVA (mg) AVG DRY WIGHT (mg) CV.8. CONG: A.9... AVG DRY B.... WIGHT (mg) % C D CV CONG: A AVG DRY B WIGHT (mg) % C D CV CONG: A AVG DRY B WIGHT (mg) % C D CV CONG: A AVG DRY B..9.. WIGHT (mg) 7% C D CV CONG: A AVG DRY B WIGHT (mg) % C D CV RMARKS: CV = (STANDARD DVIATION/MANJ'

21 Pimephales promelas FATHAD MINNOW TST. WIGHT DATA FOR LARVAL SURVIVAL AND GROWTH TST RMARKS: CV - (STANDARD DVIATION/MAN)* July. 7

22 AA# K9, FATHAD MINNOW SURVIVAL, CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHSURV. Transform: ARC SIN(SQUAR ROOT(Y) Shapiro - Wilk's test for normality D =. W -.8 Critical W (P =. Critical W (P =. ;n = ) =.97 ;n = ) =.9 Data FAIL normality test. Try another transformation. Warning - The first three homogeneity tests are sensitive to non-normal data and should not be performed. AA# K9, FATHAD MINNOW SURVIVAL, CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHSURV. Transform: ARC SIN(SQUAR ROOT(Y) Hartley's test for homogeneity of variance Bartlett's test for homogeneity of variance These two tests can not be performed because at least one group has zero variance. Data FAIL to meet homogeneity of variance assumption. Additional transformations are useless. TITL: AA# K9, FATHAD MINNOW SURVIVAL, CHRONIC, 9-8- FIL: Z:\TOXSTAT\MONT\FHSURV. TRANSFORM: ARC SIN(SQUAR ROOT(Y)) NUMBR OF GROUPS: GRP IDNTIFICATION RP VALU % % % % %.9.9 TRANS VALU

23 g, "S o, " a, "a g, *o ft. "O o. B a. ft a. TO Q, "o a. "O g, ", B g, " g, "S g, T> ft. " g, " % o. 9- " AA# K9, FATHAD MINNOW SURVIVAL, CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHSURV. Transform: ARC SIN(SQUAR ROOT(Y) STL'S MANY-ON RANK TST Ho:Control<Treatment GROUP IDNTIFICATION TRANSFORMD MAN RANK SUM CRIT. VALU df SIG 7 o, TO ft, Q, 'V % a. "o Critical values use k =, are tailed, and alpha =.

24 AA# K9, FATHAD MINNOW GROWTH CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHGR. Transform: ARC SIN{SQUAR ROOT(Y) Shapiro - Wilk's test for normality D =. W *.97 Critical W (P Critical W (P.) (n =.) (n = =.97 =.9 Data PASS normality test at P=. level. Continue analysis. AA# K9, FATHAD MINNOW GROWTH CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHGR. Transform: ARC SIN(SQUAR ROOT(Y) Bartlett's test for homogeneity of variance Calculated Bl statistic =.9 Table Chi-square value = Table Chi-square value =.9 (alpha =., df = ).7 (alpha =., df = ) Data PASS Bl homogeneity test at. level. Continue analysis. TITL: AA# K9, FATHAD MINNOW GROWTH CHRONIC, 9-8- FIL: Z:\TOXSTAT\MONT\FHGR. TRANSFORM: ARC SIN(SQUAR ROOT(Y)) NUMBR OF GROUPS: GRP IDNTIFICATION RP VALU % o, " a, "o a, " ft, " a. Ti % Q,, fi o_ TO % TRANS VALU

25 % g, T> a. TS % a. " g, " 9- " o, "o o, ^a. " * 9- a, "8 ^ T> g, AA# K9, FATHAD MINNOW GROWTH CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHGR. Transform: ARC SIN(SQUAR ROOT(Y)) ANOVA TABL SOURC DF SS MS F Between.... Within (rror)... Total 9.8 Critical F value =. (.,,) Since F < Critical F FAIL TO RJCT Ho: All equal AA# K9, FATHAD MINNOW GROWTH CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHGR. Transform: ARC SIN(SQUAR ROOT(Y) DUNNTT'S TST TABL OF Ho:Control<Treatment GROUP IDNTIFICATION 7 % % % % % TRANSFORMD MAN MAN CALCULATD IN ORIGINAL UNITS T STAT SIG Dunnett table value =. ( Tailed Value, P=., df=,) AA# K9, FATHAD MINNOW GROWTH CHRONIC, 9-8- File: Z:\TOXSTAT\MONT\FHGR. Transform: ARC SIN(SQUAR ROOT{Y) DUNNTT[S TST TABL OF Ho:Control<Treatment

26 GROUP NUM OF Minimum Sig Diff IDNTIFICATION RPS {IN ORIG. UNITS) % of DIFFRNC FROM 7 ft, * % a, % O, TJ ,

27 APPNDIX D Ceriodaphnia dubia Raw Data and Statistics

28 J^=r==:u~ Ju.k b i a Pjscharger^^^ js^.^rf-jcujcfm/ SURVIVAL AND RPRODUCTION TST I Analyst:, - DAD: Y- MAL 7

29 AA # K9 C. DUBIA CHRONIC, RPRODUCION, 9-8- File: Z:/toxstat/monte\CD. Transform: NO TRANSFORMATION Shapiro - Wilk's test for normality ******** Shapiro - Wilk's Test is aborted ******** This test can not be performed because total number of replicates is greater than. Total number of replicates = AA # K9 C. DUBIA CHRONIC, RPRODUCION, 9-8- File: Z:/toxstat/monte\CD. Transform: NO TRANSFORMATION Bartlett's test for homogeneity of variance Calculated Bl statistic =.7 Table Chi-square value =.9 (alpha =., df = ) Table Chi-square value =.7 (alpha =., df = ) Data PASS Bl homogeneity test at. level. Continue analysis.

30 FISHR'S XACT TST NUMBR OF IDNTIFICATION ALIV DAD TOTAL ANIMALS % 8 TOTAL 8 CRITICAL FISHR'S VALU (,,) (p=.) IS. b VALU IS 8. Since b is greater than there is no significant difference between and TRATMNT at the. level. FISHR'S XACT TST NUMBR OF IDNTIFICATION ALIV DAD TOTAL ANIMALS % TOTAL CRITICAL FISHR'S VALU (,,) (p=.) IS. b VALU IS. Since b is greater than there is no significant difference between and TRATMNT at the. level. FISHR'S XACT TST NUMBR OF IDNTIFICATION ALIV DAD TOTAL ANIMALS %

31 TOTAL CRITICAL FISHR'S VALU (,,) (p=.) IS. b VALU IS Since b is greater than there is no significant difference between and TRATMNT at the. level. FISHR'S XACT TST NUMBR OF IDNTIFICATION ALIV DAD TOTAL ANIMALS 7% 9 TOTAL 9 CRITICAL FISHR'S VALU (,,) (p=.) IS. b VALU IS 9. Since b is greater than there is no significant difference between and TRATMNT at the. level. FISHR'S XACT TST NUMBR OF IDNTIFICATION ALIV DAD TOTAL ANIMALS % TOTAL CRITICAL FISHR'S VALU (,,) (p=.) IS. b VALU IS Since b is greater than there is no significant difference between and TRATMNT at the. level. SUMMARY OF FISHR'S XACT TSTS NUMBR NUMBR SIG

32 GROUP IDNTIFICATION XPOSD DAD (P=.) % % % 7% % TITL: AA # K9 C. DUBIA CHRONIC, RPRODUCION, 9-8- FIL: Z:/toxstat/monte\CD. TRANSFORM: NO TRANSFORMATION NUMBR OF GROUPS: RP IDNTIFICATION g, " Q, " a, "S a. " a. ^a, Ts a, " Q, ~. " a, " o, T> O, Tb a, " g. " g, Ti Q, "o a. TJ, " g, 9- «o, o o, "S a, "o o, "o - T>. Q T) O, TJ RP VALU TRANS VALU

33 g, o a. "o, "a a, " g. " g, "9 g, " Q, % o, "8 g, i, "O g. " o. "o 9- ", " ft. "S g. Q, "V O, Ti a. "S, * AA # K9 C. DUBIA CHRONIC, RPRODUCION, 9-8- File: Z:/toxstat/monte\CD. Transform: NO TRANSFORMATION ANOVA TABL SOURC DF SS MS F Between 9, Within (rror) Total 9., Critical F value =. (.,,) Since F > Critical F RJCT Ho: All equal AA # K9 C. DUBIA CHRONIC, RPRODUCION, 9-8- File: Z:/toxstat/monte\CD. Transform: NO TRANSFORMATION DUNNTT'S TST TABL OF Ho:Control<Treatment ROUP IDNTIFICATION TRANSFORMD MAN MAN CALCULATD IN ORIGINAL UNITS T STAT SIG 7 o. "S % g, " q, g,

34 Dunnett table value =. ( Tailed Value, P=., df=,) AA # K9 C. DUBIA CHRONIC, RPRODUCION, 9-8- File: Z:/toxstat/monte\CD. Transform: NO TRANSFORMATION DUNNTT'S TST TABL OF Ho:Control<Treatment GROUP NUM OF Minimum Sig Diff IDNTIFICATION RPS {IN ORIG. UNITS) % of DIFFRNC FROM 7 9- * g, " a, " % g, "S.,....,, , ,. AA # K9 C. DUBIA CHRONIC, RPRODUCION, 9-8- File: Z:/toxstat/monte\CD. Transform: NO TRANSFORMATION STL'S MANY-ON RANK TST Ho:Control<Treatment GROUP IDNTIFICATION TRANSFORMD MAN RANK SUM CRIT. VALU df SIG 7 Q, " a. " q, *, "o, " 8..,.,. 7..8,, 9,, 8.. 7,, 8.. 9,,, ,,,.... Critical values use k =, are tailed, and alpha =.

35 APPNDIX Organism History

36 AQUATOX,INC. TWIN POINTS ROAD HOT SPRINGS, ARKANSAS TST ORGANISM HISTORY DAT SHIPPD CLINT Purchase Order #: SPCIS: (Pimephalcs promelas> Mysidopsis bahia Cyprinodon variegates Quantity Shipped: Age: t* I/? J& Brood Stock Source: Culture Water: <^'&roundwater } Artificial Salts Artificial Salts Hardness (Mg/ CaCO^ Salinity (ppt) Dissolved Oxygen (Mg/): / ' / Feeding: Comments: Shipped Via: UPS Overnight Packaged By: Bill Hall Prinlers7

37 Blue Spruce Drive, Suite C Fort Collins, Colorado 8 Toll Free: 8/-9 Tel:97/8-9 Fax:97/8- ORGANISM HISTORY DAT: //9 SPCIS: AG: LIF STAG: HATCH DAT: BGAN FDING: FOOD: Ceriodaphnia dubia Variable Adult Variable Immediately YTCjjSelenastrum sp. Water Chemistry Record: Current Range TMPRATUR: SALINITY/CONDUCTIVITY: TOTAL HARDNSS (as CaCO): TOTAL ALKALINITY (as CaCO): ph: C mg/l mg/l C 8-m/l - ma/ Comments: Facility Supervisor Aquatic BioSystems, Inc Quality Research Organisms

38 APPNDIX F Quality Assurance Charts

39 ARKANSAS ANALYTICAL, INC. FATHAD MINNOW SURVIVAL QUALITY ASSURANC ro u '* I" D. - Q) So n V^f _^^ >^L -NOC UCL? Date Test Performed A RKANSAS ANALYTICAL, INC. FATHAD MINNOW GROWTH QUALITY ASSURANC e Toxicant,,ppm Referenc KC 8 ' NOC UCL LCL Date Test Performed

40 ARKANSAS ANALYTICAL, INC. CRIODAPHNIA DUBIA SURVIVAL QUALITY ASSURANC 'X K a * _r c O I * 'ai n -» - > + ^r^ N.^/ ^^»»» ^^^ - - " UCL Date Test Performed ARKANSAS ANALYTICAL, INC. CRIODAPHNIA DUBIA RPRODUCTION QUALITY ASSURANC Reference Toxicant, KCI, ppm 7 -I ' - f^ ^^ ^^~y^^ ^ ^ - _^^. _ ^,^ i * NOC Date Test Performed