USA, Inc. March 28, 2007 Sludge Solutions International, LLC Sewper Rx (granular host) YO60206C: Day Static-Renewal C. dubia & P. promelas Definitive Toxicity Results Bioprocessing LLC 5041 Taravella Rd, Marrero, LA 70072 Altention: Boyd Young Dear Mr, Young: ‘A pair of 7-day chronic C. dubia and P. promelas detintive tests (EPA-821-R-02-013) was initiated March 12, 2007 (Appendix A). The sample used in this test was supplied by Bioprocessing LLC on behalt of their align Sludge Solutions International, LLC and delivered to Environmental Enterprises USA, Inc. (EE USA) on March 8, 2007 (Appendix D). Daily, a solution of Sewper Rx (granular host) was propared at 29.88 mg/L, the recommended application rate (402 per 1000 gallons water), and mixed for one hour ona stir plate ‘The C. dubia test included ten replicates each of Sewper Rx (granular host) at 29.96 mg/L and a concurrent laboratory performance control (LPC). The P. promelas test included five replicates each of Sewper Rx (granular host) at 29.96 mg/l. and a concurrent LPC. Moderately Hard Synthotic Freshwater was used as the diluent and LPC. C. dubia were fed 0.1ml Solenastrum capricornutum and 0.1m\ of Yeast-Cerophyi-digested Trout Chow daily and P. promelas test organisms were fed Artemia nauplii less than 24-hours old twice daily Statistically Significant Difference belween LPC and EER: ‘Sewper Rx (granular host) at 20.96 mg/L. E1G207 Survival Reproduction Growth ©. dubia No No | NA P. promelas No NA No 58485 Pearl Acres Road ~ Suite D — Slidell, Louisiana 70461 OFFICE: §04-646-2787 — FAX: 504-646-2810 ~ 1-800-986-2788 Page 1 of 2The response used in statistical analysis of survival data was the proportion of surviving test organisms per replicate. The response used in reproduction data analysis (C. dubia) was the total number of neonates produced per replicate. The response used in growth data analysis (P. promelas) was the average individual dry weight for each replicate: replicate weight divided by the number of orginal larvae. The Homoscedastic {Test was used to determine if there were statistically significant differences in the endpoints of interest, survival and reproduction for C, dubia or survival and growth for P. promelas, between the LPC and 29.96 mg/L Sewper Rx (granular host). In these tests, survival and reproduction of C. dubia and survival and growth of P. promelas were not significantly reduced alter exposure to 28.96 mg/L Sewper Rx (granular host). The results for the laboratory perfomance control and Sewper Rx (granular host) were very similar. Copies of the raw data are presented in Appendix A. Survival, reproduction and growth data analysis and summary statistics for the C. dubia and P. promelas tests are presented in Appendix B & C, respectively. Mr. Young, thank you for choosing EE USA to complete this work for Bioprocessing LLC. | appreciate the opportunity and look forward to working with you again. Please call me if you have any questions or comments. David L. Daniel Laboratory Director Muh Oi! 3.29 (01 Mark A. O'Neil QN/QC Supervisor encl: Appendixes A ~ D Page 2 of 2Environmental Enterprises USA, Inc. APPENDIX A7122007, 346 PM Environmental Enterprises USA, Ine processing LLC Boyd Yound Test Concentrations, mg/I Product (PR) Ceriodaphnia | Pimephales | Total Volume! J Color] mi mi dubia yromelas_| Concentration, ml DH,O 29.96 mg/l 1500.00 0.00] 0.0 mgit LPC Total Volume (mil) of EFF needed per day= 1500.9} C. dubi ‘Total Volume (ml) of EFF needed for test duratior {____Total Volume (ml) of EFF needed for test duration= _10500.0} PR Stock Solution pippared ean gallons = 44.94 mg SewperIX 1300 mf DAO. fo g Da Fags & Cacao 1 as HATA Cheek by: SW ALN 30 ml = 1000 ml 40500, (0 Reps x 15 ml P. promelas = 4 Reps x 250 ml DI1,0 = Dilution Water = Moderately Hard Synthetic Freshwater Alkali UPC | My LPC TPC | M fe | Date Oana | “056 Alkalinity Te [= Fite [=| G8 [Conductivity [293 InSl273 [nel 242 || Hardness | 100_|~ yoo | ~ [yo |= pir 4.2 W192 [RB[ 9.7 [ee ‘TRC a [it | | L- = L ha pH: SU TRC: mg/l M#: Meter number LPC: Laboratory Performance Control, symthetie moderately hard water ity: mgflas CaCO, Conductivity: jS/em Hardness: mg/l as CaCO, ee __ Prep Date’ L 03/12 03/13, 03/14 03/16 03/17, 03/18 Weight |W 9F Te a7 | 9G eG | oree4 [Ge G4 Initial Ve mM VER Sie & Se Comments: Lot 2syi007 386 9M Environmental Enterprises USA, Ine Cladoceran, Ceriodaphnia dubia Survival and Reproduction Test Method 1002 Bioprocessing LLC —oSSS Test O: 22 Hours Old Test Organisms Sou Test Initiation At: _(@22 _ onz3,//2 107 Counted by: ec A 2 _ QCIQA by: Vas Loaded by: Bey C. dubia Daily Survival Data Treatment: Laboratory Performance Control (LPC), 0 mg/l wie TT 23 Tey Sees eer REP Suv. | Neontes | Teck Per Day TOO eae po D 1 | We OO lelolelelololely lwo Ke “PF lolo lololelolelolofohesl co fe — "PF lob fo[3lole lo lo lols lao aig nel TION fo FIol¢ [4h [ou fio! Gul * halishalicheh3halatirfo fio lay [se * | [alshobalsal adn | ighos [aa 6 7 3rd Breed Reprodn For RETA Wear) OV OT [ 2u| 35] 31/3 44s [34134 ]33 [as |24 [auc Comments: E-132-07 ona37122007, 3:46 PM Environmental Enterprises USA, Inc C. dubia Daily Survival Data Cont. Tink ‘Treatment: 29.96 ms I 12 [ 13] 4] 15 | 16 | 17 | 18 | 19 | 20 | % No. of Re Bar | Neowtes | tech | tine TTT epee ey pe Lace > foalolololelslololale lal o kylite “ P lelelelelolols lo lola fisol oc fre lasy "Plo le bb Jol3lFfoldelohodlio Ire hsud ‘ bal lalel oh |g lalate vr |-relizag * T2018 els lis fir is [ry fe [ov [94 | Sdizos © We hBaliaeligliajo lie ho |< [Se [lis 7 Sr Bron RrodTon PRT ew eV 95 lub |Yajul [a4 4] ag]a3 bo [37 a6 4 3-107 / Se Calculations thaw pohe onac ty OF Data Entry by: Us Ne 2B Yh Double Data Entry by: a eV or QA/QC Officer: ni E-132-07 NOECILOEC 30f 1237122007, 3:6 PM Environmental Enterprises USA, Inc. CC. dubia Water Quality Data LPC & AU Treatments: Initial & Final Temp. ~ 23.5 ~ 26.4°C; Initial & Final DO ~ 4.028.3 mg/L Dao TTT 03/1207 LPC a | Mee "143 | 3 31 r | 7317.5 s7 te" [aya |as.0 ag * [83/5 1 Qs mt Tse] sg | Os initials Times Thital Time Final Timer DayT Treatment mg/l PR oB/is07_| LPC] 39.96 Meter mel # DO ies 3:2 : 1% | 7.4 a Temp | eat | Sse xm = Se nO} gae7 B83 | ASH i ® Flegl sd initials “This PAE yn. Limes [atime soo Day? oaanT | FE] 3956 Meter gil é ‘po 7 De "Tas A 4-9/7 Temp : 995.1 (9S.9 yu7 [pet pH Vv EL Initials, Initials: Wwe Finals: ‘Times [ial Time: 435@ Final Time) © Date rn01 0 wvreled oafeqpr U- B-132-07 NoxenOEC 4of 1237122007, 3.46 PM Environmental Enterprises USA, Ine. C. dubia Water Quality Data Cont, Day3 “Treatment mgt PR aster [LPC [29.96 Meier mg/l # 114 | 4.0 51 La2|8% | Sal 9544 |2>5 ms el [tS Ind Bz = ag Taltals Tnilals Fins 50) AD Times, tial Time: 1450 Final Time: 15 Bayt Teameat mat PR 03/16/07 LPC 29.96 ~] Meter mg/l # we Teste 7 “133 le3 37 fer [194.4 15G NS * bes3 [254 ag mF Lso]| Zo ae Initials Finals: G Times. we Final Tis =F Tamara 03/17/07 LPC 29,96 Meter: oral A me '([s3lsi s2 |" {23 |g. Sal te | |26a|o26 245 |23.% Mm pF YL so | 50 Yh Totals i nue Finals, Se "Times Initial Time: [VS Final Time: | SO E-132-07 NOECILOEC Sof 2« WI22007, 346 PM. Environmental Enterprises USA, Inc. C. dubia Water Quality Data Cont. Dayé ‘Treatment mg/l PR wiso7 | LPC | 29.96 Meter mg/l # 4 7 DO Tem T 23 @ Me LPC: Laboratory Performance Control, symhetic moderately hard water Alkalinity: mg/l as CaCO; Conductivity: ytS/em — Hardness: mg/l as CaCO; pH: SU TRC: mg/l Comments: E-132-07 NOECLOEC 6orl2