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Home My WebLink AboutWI0501074_More Information (Received)_20210623SiREM Leading Science•Lasting Solutions 23 June 2021 Mike Burcham Geosyntec 11490 Westheimer, Suite 150 Houston, TX 77077 mburcham(c�geosyntec.com Reference: Low pH KB-18Plus Culture Dear Mike: 130 Stone Rd. W Guelph, ON IV1G 3Z2 4519) 822.2265 Low pH . KB-1® Plus (LpH KB-1® Plus) and KB-1® bioaugmentation cultures introduce key microorganisms to contaminated sites where they are absent or in low concentrations. Bioaugmentation with LpH KB-1® Plus and/or KB-1® is an effective solution to enhance remediation of chlorinated ethenes, with the primary organism being Dehalococcoides in both cultures. Many sites have an intrinsic pH below the optimum range for optimal bioremediation of chlorinated solvents (pH 6.5-7.8). For example, at a pH below 6.0, slow and incomplete dechlorination of chlorinated ethenes is often observed, leading to accumulation of cDCE and VC. SiREM's LpH KB-1® Plus culture is acclimated to dechlorinate chlorinated ethenes at a lower pH. The production of LPH KB-1® Plus is almost identical to that of our KB-1® culture, with the pH range the cultures are maintained at being the primary difference. The following steps provide an overview of SiREM bioaugmentation culture production from initial growth to shipment of the completed product to a site: 1. Stainless steel growth vessels are cleaned using soap and water and filled with reverse osmosis (RO) purified water (SOP #003). 2. Sterile media concentrates (with the exception of bicarbonate, ferrous sulfide and vitamins) are added to the RO water. 3. Vessel containing water and non -heat sensitive media components are sterilized using pressurized steam (SOP #067). 4. The vessel is purged with sterile filtered 80% nitrogen / 20% carbon dioxide gas mixture to remove oxygen that otherwise would kill anaerobic microorganisms (SOP #005). Heat -sensitive concentrates (bicarbonate, FeS and vitamins) are added after steam sterilization. Bicarbonate and FeS are added aseptically from smaller (1 L) vessels. Vitamins are added aseptically by addition through a sterile 0.45 pM filter. 5. The pH of the media is checked and adjusted as necessary. (i.e., for KB-1® culture pH range of 6.8 — 7.5 and for LPH KB-1® Plus pH range is 5.5 -6.0) Mike Burcham 23 June 2016 Page 2 of 2 SIREM 6. Sterility of completed media is confirmed by Tryptic Soy Broth (SOP #059). If media passes the test, the vessel is approved for inoculation. 7. A 15% to 50% inoculum by volume (15 to 50 liters in a 100 L vessel; 1.5 to 10 liters in a 20 L vessel) of the required bioaugmentation culture is transferred by gas pressure from the seed culture vessel through sterile hoses into the sterile media (SOP #004). 8. The culture is amended with required electron acceptor (e.g., TCE for KB-1® and LPH KB-1® Plus) to provide growth substrate for the culture and to ensure degradation activity is maximized and that key microorganisms reach required concentrations. 9. DNA is extracted from the culture for molecular genetic analyses monthly. Concentrations of key degrading organisms (i.e., Dehalococcoides) are monitored by qPCR. 10. Once degradation activity, microbial composition, and key degrader concentrations are demonstrated to meet specifications, the product is ready to be transferred to shipping containers. Shipping containers are 100 L or 20 L stainless steel vessels, which are steam sterilized prior to transfer. 11. The shipping containers are purged with an 80% nitrogen / 20% carbon dioxide gas mixture ensuring the bioaugmentation cultures are transferred into the container without exposure to oxygen. The gas mixture is used to pressurize the growth vessel and push the culture through sterile tubing into the shipping container. 12. The shipping container is purged with 80% nitrogen / 20% carbon dioxide gas mixture to remove volatile compounds and gases. The absence of significant concentrations of chlorinated ethenes is confirmed by gas chromatography prior to shipping. 13. Final quality control checks including key degrader concentrations by qPCR, pH, and biodegradation ability of the purged culture are performed. 14. The shipping container is placed into a protective over -pack and shipped to the injection site by express courier (SOP #013). Growth media is prepared following the same SOP for all SiREM anaerobic bioaugmentation cultures. The media composition for LpH KB-1® Plus and KB-1® is provided in Attachment 1. The pH in the LpH KB-1® Plus culture is allowed to decrease naturally and sodium bicarbonate is added as required to maintain a pH in the 5.5-6.0 range. The pH in the KB-1® culture is maintained in the 6.5-7.0 range with the addition of sodium bicarbonate. We trust that this information will meet your needs at this time. Please feel free to contact me at 519-515-0840 if you have any questions or you require additional information. Sincerely, Jeff Roberts, M.Sc. Operations Manager siremlab,com iLit;' Attachment 1: KB-1® & Low pH KB-1® Plus Media Composition s renilab.com Attachment 1: KB-1® & Low pH KB-1® Plus Media Composition SiREM Leading Science • Lasting Solutions Ingredient Composition in product (%) Potassium Phosphate Dibasic 0.027 Potassium Phosphate Monobasic 0.034 Ammonium Chloride 0.053 Calcium Chloride 0.007 Magnesium Sulfate 0.0125 Ferrous Chloride 0.002 Sodium bicarbonate 0.2 Ferrous Ammonium Sulfate 0.04 _ Sodium sulfide 0.012 Resazurin 0.0001 Boric Acid 0.00006 Zinc Chloride _ 0.00002 Sodium Molybdate 0.00002 Nickel II Chloride 0.00015 Manganese Chloride 0.0002 Copper II Chloride 0.00002 Cobalt Chloride 0.0003 Disodium Selenite 0.000004 Aluminum Trisulfate 0.00002 Vitamins _ 0.001 Water 99.61061 Total 100.00 ATTACHMENT C: Chemical Components in KB-1® Growth Media Chemical Name Chemical Formula CAW/Purpose Final Concentration (grams/liter) % Product Composition Potassium Phosphate Dibasic KH2PO4 11/4/7758 pH buffer 0.27 0.027 Potassium Phosphate Monobasic K2HPO4 7778-77-0 pH buffer 0.34 0.034 Ammonium Chloride NH4CI 12125-02-9 mineral for microbial proliferation 0.535 0.0535 Calcium Chloride CaCl2 10035-04-8 mineral for microbial proliferation 0.07 0.007 Magnesium Sulfate MgSO4 10034-99-8 mineral for microbial proliferation 0.125 0.0125 Ferrous Chloride FeCl2 13478 mineral for microbial proliferation 0.02 0.002 Sodium bicarbonate NaHCO3 144-55-8 mineral for microbial proliferation 2 0.2 Ferrous Ammonium Sulfate (NH4)2Fe(SO4)2 7783-85-9 mineral for microbial proliferation 0.4 0.04 Sodium sulfide Na2S 1313-84-4 mineral for microbial proliferation 0.12 0.012 Resazurin C12H6NNa04 62758-13-8 mineral for microbial proliferation 0.001 0.0001 Boric Acid H3B03 10043-35-3 mineral for microbial proliferation 0.0006 D.00006 Zinc Chloride ZnCl 7646-85-7 mineral for microbial proliferation 0.0002 0.00002 Sodium Molybdate Na2Mo04 10102-40-6 mineral for microbial proliferation 0.0002 0.00002 Nickel II Chloride NiC12 7791-20-0 mineral for microbial proliferation 0.0015 0.00015 Manganese Chloride MnCl2 13446-34-9 mineral for microbial proliferation 0.002 • 0.0002 Copper II Chloride CuCl2 10125-13-0 mineral for microbial proliferation 0.0002 0.00002 Cobalt Chloride CoCl2 7791-13-1 mineral for microbial proliferation 0.003 0.0003 Disodium Selenite Na2Se03 10102-18-8 mineral for microbial proliferation 0.00004 0.000004 Aluminum Trisulfate Al2(SO4)3 10043-01-3 mineral for microbial proliferation 0.0002 0.00002 Biotin (Vitamin B7) C16H16N203S 58-85-5 nutrient for microbial proliferation 0.000002 0.0000002 Folic Add (Vitamin B9) C19H19N706 59-30-3 nutrient for microbial proliferation 0.000002 0.0000002 _ _ _ Pyridoxine HCI (Vitamin B6) C8H„NO3 65-23-6 nutrient for microbial proliferation 0.00001 0.000001 Riboflavin (Vitamin B2) C17H2ON4O6 83-88-5 nutrient for microbial proliferation 0.000005 0.0000005 Thiamine (Vitamin B1) C•2H.7N4OS' 70-16-6 nutrient for microbial proliferation 0.000005 0.0000005 Nicotinic acid (Vitamin B3) C6NH502 59-67-6 nutrient for microbial proliferation 0.000005 0.0000005 Pantothenic acid (Vitamin B5) C21H36N7016P3S 85-61-0 nutrient for microbial proliferation 0.000005 0.0000005 p-Aminobenzoic Acid (Vitamin B10) C7H7NO2 150-13-0 nutrient for microbial proliferation 0.000005 0.0000005 Cyanocobalamin (Vitamin B12) C63H66CoN14O14P 68-19-9 nutrient for microbial proliferation 0.000005 0.0000005 Thioctic (lipoic) acid C5111402S2 1200-22-2 nutrient for microbial proliferation 0.000005 0.0000005 Coenzyme M C2H503S2 3375-50-6 nutrient for microbial proliferation 0.0001 0.00001 Dehalococcoides Spp. - - micobial community - 0.0007 GeobacterSp. - - micobial community - 0.0005 Methanomethylovorans Sp. - - micobial community - 0.02 Water H2O 7732-18-5 matrix - 99.59 TOTAL 100 v ATTACHMENT C: KB-1°Plus Ingredient List Chemical Name Chemical Formula CAS# Purpose Final Concentration [grams/liter) %Product Composition Potassium Phosphate Dibasic KH,PO4 11/4/7758 pH buffer 0.27 0.027 Potassium Phosphate Monobasic K2HPO4 7778-77-0 pH buffer 0.34 0.034 Ammonium Chloride NH,CI 12125-02-9 mineral for microbial proliferation 0.54 0.0535 Calcium Chloride CaCl2 10035-04-8 mineral for microbial proliferation 0.07 0.007 Magnesium Sulfate MgSO4 10034-99-8 mineral for microbial proliferation 0.125 0.0125 Ferrous Chloride FeCl2 13478 mineral for microbial proliferation 0.02 0.002 Sodium bicarbonate NaHCO3 144-55-8 mineral for microbial proliferation 2 0.2 Ferrous Ammonium Sulfate (NH,)Fe(SO,la 7783-85-9 mineral for microbiaproliferation 0.4 0.04 Sodium sulfide Na- S 1313-84-4 mineral for microbial proliferation 0.12 0.012 Resazurin Cr2HeNNaC4 6275913-8 mineral for microbial proliferation 0.001 0.0001 Bonc Aad H3B0) 10043-35-3 mineral for microbial proliferation 0.0005 0.00006 Zinc Chloride ZnCI 7646-85=7 mineral for microbial proliferation 0.000.2 0.00002 Sodium Moivtdate NaLMo04 10102-46-6 mineral for microbial proliferation 0.0002 0.00002 Nicarel II Chloride NiC12 7791-20-0 mineral for microbial proliferation 0.0015 0.00015 Manganese Chloride MnCl2 13446-34-9 mineral for microbial probreration 0.002 0.0002 Copper II Chloride CuCI; 10125-13-0 mineral for microbial praltferau❑n 0.0002 0.00002 Cobalt Chloride CoCI, 7791-13-1 mineral for microbial proliferation 0.003 0.0003 Disodium Setenite Na2Se02, 10102-18-8 mineral for microbial proliferation 0.00004 0.000004 Aluminum Trisutfate Al2(SO4)3 10043-01-3 mineral for microbial proliferation 0.0002 0.00002 Biotin (Vitamin B7y C,,,H,6Ni03S 58-85-5 nutrient for microbial proliferation 0.000002 0.0000002 Folic Acid (Vitamin B9) CI9Hj9N; Oe 59-30-3 nutrient for microbial proliferation 0.000002 0.0000002 Pyridoxine HCI (Vitamin B6 CBH„NO3 65-23-6 nutrient for microbial proliferation 0.00001 0.000001 Riboflavin {Vitamin B2y C17H20N406 83-88-5 nutrient for microbial proliferation 0.000005 0.0000005 Thiamine (Vitamin B1) C;,H, �N3OS' 70-16-6 nutrient for microbial proliferation 0.000005 0.0000005 Nicotinic add (Vitamin B3y C.,NH50= 59-67-6 nutrient for microbial proliferation 0.000005 0.0000005 Pantothenic acid (Vitamin B5) C2,H.N7O,6P.S 85-61-0 nutrient for microbial proliferation 0.000005 0.0000005 p-Aminobenzoic Acid (Vitamin B10) C7H: NO- 150-13-0 nutrient for microbial proliferation 0.000005 0.0000005 Cyanocobalamin Vitamin B121 C63HeeCoN,4014P 68-19-9 nutrient for microbial proliferation 0.000005 0.0000005 Thioctic (fipple} add CjHt&O2_S_2 1200-22-2 nutrient for microbial proliferation 0.000005 0.0000005 Coenzyme M C2HiO362 3375-50-6 nutrient for microbial proliferation 0.0001 0.00001 Dehalococcoides Slur . - - micobial community - 0.0004 Geobacter Sp. - - micobial community - 0.0002 Methanomethylovorans Sp. - - micobial communes - 0.007 Dehalobacfer Sp. micobial community 0.0002 Dehalogenimonas Sp. micobial community 0.00008 Water H2O 7732-18-5 matrix - 99.603 fi TOTAL 100 dehalobacterand dehalogeminoas are NOT included in the KB-1 Plus formulation that is proposed for use (those are used for KB-1 Plus formulations that are degrading other chlorinated solvents like 1,1,1-TCA or chloroform 2020 Sampling Summary Allegion Rocky Mount Rocky Mount, North Carolina Monitoring Well ID Aquifer Monitored Injection Ares Perform.. Monitorbrr Analysis Natalia Organic Compounds Met u a. Mena, end Ethane Total Organic Carbon (TOO. Defaniococrddes and reductive dehalogewe (motional genet (IreA,6srel, end verA) Sulfate Sulfide Volatile Fatty Acids(VFAs) Iron and Manganese (Nat in Bien IMWP) TDS (fiat In Blo I8MW15 Nitrogen, Nitrate (Not In �o 1MWP) EPA Method 8260D EPA Method RSK-175 SM 53I0C GeneTrei EPA Method 9056A SM 4500 EPA Method 8015D EPA Method 6010D SM 2540 EPA300/ 9056A Sampling Month: Nov-20 Nov-20 Nov-20 Jun-20 Nov-20 _ Nov-20 Nov-20 Nov-20 _ Nov-20 _ Nov-20 High Concentration Injection An (MCI, HC2, HC3, HC4) MO, •;I i) I HY. -Treatment Area X 7: t•_ - X X X MW-76S M HI 1-Treatment Area X X ' .. I - _ 0 X X MW46D L HCI- Dawnedien g DG3-H., la Jientt Au 2020 g - - - - - - r - - - MWA6S M HC1-Downgredrent HC2-Dowry.'....",ipra X _ - - _ - - _ _ - MW-75D L HC2-Downward Mntetion X - - - - - - - - - Mu-ISf M zit'2-Treatment Area X - - - - - - - - - MWv H 1IL_ - I,xn.rm Ib-_ids-. X - - k - - - - - - MW''BD L Ht. 3-Treatment Area X X X X X - - X X X /(W.'aS M Ht. "3-Treatment Area X X X X X - - X X X MW-58S M HC3-Tresmmnt influence Feb2021 - - - - - - - - - MW-77D L HC3-Downgrulient X - - - - - - - - - MW-77S M HC3-Dowryrrlient X - - - - - - - - - MW-30 L HC4-TreementArea X - - X - - - - - - Dosr client Injeedon Arne (DGI, DC2, DG3) MW-37S M DGI- Treommnt Area X X X X X - - X X X MW-47D L DUI -Treatment Area X - - - - - - - - - MW48D L DG1 - Down.,,,: iea _ _ _ - MW-74D L DG2- Downgrediem X - - X - - - - - - L DG3-Uswli - -MW-70D - - - - MW4IDDR L DG3-O.+11yoimrl X X X - X - - X X X 1 oat Wear J.• MW440 L Downgredient-Sentinel Aug2020 - - - - - _ - - - MW44S M Downmadient-Sentinel Aug 2020 _ _ - _ - _ - - - MW-72D L Downgmdient-Sentinel Aug 2020 - - - - - - - - - MW-72S M D0000 &ent-Sentinel Au; 2020 - - - - - - - - - I. DN. proem MOM mmirigxtW included in the Bimemedmum Wain, Memme Was Panfor+pe mmpe•m6r4w'ryrae lmw aNaraiW3mmamp4'$ermb. 2. X indinaepnuMmnv>®plea were collected duirsthe month wdiatl, on. W spec. ors Mymeeffe most roc.mmptig EMI a provided. 3.-'udicma w Os wag on eampkd for am mdimtd compounds. 4. Aquifers mmitad MdeM for Mille AquifermdL for Inver Ag f0. 5. EPA indicate untied Stan 6nv'uwn meal Pmaeam Agermy. 6.. indiums smrdend mould. 2021 Bioremediatiea Interim Measures Wodplen Page 1 of 1 Ceosynteco autsullants Table 5. Bioremedialion Performance Monitoring Plan REVISED AIlegion Rocky Mount Rocky Mount, Nord] Carolina Monitoring Well ID Aquifer Monitored Injection Area a Performance Monitoring Analysis Select Volatile Organic Compomdsl Methane, Ethane, and Ethane Told Organic Carbon (TOC) Uelwfococmides and reductive delWngenase functional genes (lreA , bvcA, end A) Sulfate SulSde J Volatile Fatty Acids (VFAs) EPA Method 8260D EP�AKMeI17h5°d SM 5310C Genie-Tmc EPA Method9056A SM4500 EpA�MDod Year I Year Yearl I Year2 t Year1 r Year2 Year 1 Year Year I Year Yearl I Year Yearl 1 Year High CmcenMtlon action Areas (Ha, HC2, HC3, HC4) MW-76D L HCI - Treatment Am 05 S 3/5 8 5 S 3 3 QS 5 Q3 Pending Remus MW-76S M HCI-Trmmmt Ares Q5 6 iFF S 5 S 5 5 Vb S gin. 4 Fading Results MW-46D L HC13Downgmdl.1t A A A A A A - - - - - - Pending Results MW-46S M CI - Ibwagmthem HC2-G,..,, ,.sYend A A A A A A - - - - - - Pending Rmha MW-75D L Hi: - Downward Mn,rgmi 5 A A A A A A A - - - - Penang Romb. 1164 •5. M Hl:-Treatment Area .}S S / Q5 S S S S 3 SS Sae • noxE,Rawn M1lr� M HC2-Tr -Treatment Boundary 5 5 S 5 S S 5 A g3 Q'=s Maims NaomiMW Mw.1'A,1 1 HC3 •Tmamma .wm Ir5 5 C,.$ 5 S S 5 3 Qa -. hadmg kawn• Mw-Tag M Hat -r,tLAtm (3.5 5 Q3 S S S 5 5 U'S lI5 x Vends, ltrin: I. MW-51S M ]ff l-iersvnene OiRueue UnaJatl A A A A A A - - - - - - Paling Results MW-77D L HC3-Dox ow ...Vent A A A A A A - - - - - - Peeling Results MW-773 M HC3-Domtgmdiem A A A A A A - - - - - - Pending Results MW-30 _ L HC4- Treatment Area .)5 S ;/S 5 S S S _ 3 _ i; 3 S .: 5 fending Results Dmng.adimt Infection Areas (DG1, DO2, DG3) MW-37S M DG1-Treatment Am Q/S S QIS 5 F 5 S 5 5 Q(S S Q/S 5 Pendmg Results MW47D L DGI-TmetmentAm V S S Q/S S S S 3 S V s S s,1 5 • Pending Results MW48D L DGI-Downgmdimt S S S S S S - - - - - - Fading Results MW-74D L DG2-I6••nor.,,6mt :•8 5 v/S 5 u/S S 5 S Q/S S QIS Pending Results MW70D L DG3-Ugndient µS 5 QIS S QIS S 5 S Q/S S V/S S Pending Results MW4IDDR L DG3-Domµnd lent J .; S S s /S S 418 3 S S Q/S S Q/S • fading Results S.M.! 55545, MW44D L Domigradient-Sentinel Vl re, A - - QI:-Qt A - - ..QI/42r A QI!'_� A - - MW44S M Doxny-.dine- Sentinel .1l'U1 A - - 1211Q4 A - - 01/04 A QI/Q4 A - - MW-72D L Downgmdiem-Sentinel L'I Q4 A - - QI/QI A - - QI/Q4 A GINA A - - MW-723 M Do..,Y,ndient-Sentinel •31 Ql A - - a1 f94 A - - D1/Ut A Q1/I.. A - - Total Analytical Samples rmYear .- 36 43 32 43 36 25 2A 44 29 44 28 Pending Results so I. Seta roMde optic cmp: eb(V06) hide termhloroc ne(PM nielJax4cr Rf=air•St.Dirhbatar, own -I,14)Ce,1.lditlJvnYeq vinyl Wed* and meJyl ethyl beme N.} S. Aquifers mamaedksbnie M fon Adage Aquifer .ML hLoxrr Apdfer. 3.FmgmryafpeR erne mailed. ud Led below s. Q5 ininepm¢r5th s®iovmlmmpW.(i.e. Madl3. Math,.MMmh 12film*. Iledimm). b. S India. snmianroalsmpW.O.. Stath 6 end Madl1l2 adman *aim). e. A i.e..es mud amyl:. (i e. Marsh 12 known. inaction). d- i4i k. de pameaaAlmbeiridd Emile pm! a mvwa'gpblfa Ya even year. e. Pend. Rea. Winans nal o13tia1mMod my be anpanonaeddmi.eevms.,, depvtivg mW erOdnal nopW.mads a4Wmed LQI /Q0 Wks. Mate 3 dun Math 12 ansplireollawi.*eda . 4.EPAbdict aUKW SWaFirvhammhlPMatia,ASnry. S. MA indimo+madmd method. 2021 Binmmediation Interim Measures Workplan Page l oft GeosyntecD consultants