Res 056-2011 6/21/2011 RESOLUTION NO. 56-2011
Resolution Authorizing The City Manager To Execute All Documents
Necessary To Purchase Biocide Screening, Optimization, And Pilot
System Testing Services For The Mitigation Of Microbiologically
Influenced Corrosion Of 316L Stainless Steel Within The Cypress
Water Treatment Micro filtration Plant, From Ecolyse, Inc., In An
Estimated Amount Of $90,407.73
WHEREAS, the City of Wichita Falls desires to control the microbiologically
influenced corrosion of the 316L Stainless Steel components within the Siemens
Microfiltration Plant;
WHEREAS, after extensive search, the City has found only one testing firm
capable of providing biocide screening, optimization, and pilot system testing services
for the mitigation of microbiologically influenced corrosion of 316L stainless steel within
the City's Siemens microfiltration plant;
WHEREAS, the City Council finds that the aforementioned biocide efficacy
testing and optimization services are professional services and are exempt from Texas'
requirement for competitive bidding;
WHEREAS, the City Council finds that the biocide efficacy testing and
optimization services proposed to be provided by Ecolyse, Inc. are also exempt from
Texas' competitive bidding requirements of Chapter 252 of the Texas Local
Government Code because they are the sole source of said services.
NOW, THEREFORE, BE IT RESOLVED BY THE CITY COUNCIL OF THE CITY
OF WICHITA FALLS, TEXAS, THAT:
The City Manager is authorized to execute all documents necessary to purchase
biocide screening, optimization, and pilot system testing services for the mitigation of
microbiologically influenced corrosion of 316L stainless steel within the Cypress Water
Treatment microfiltration plant in an estimated amount of $90,407.73, from Ecolyse,
Inc., in accordance with the attached Proposal, in a form approved by the City
Attorney.
PASSED AND APPROVED this the 21st day of June, 011.
MAYOR
TTEST: '
`
ity Clerk
1HIS DOCUMENT INCLUDING, DRAWINGS, PROCEDURES, SPECIFTCATIONS, AND TfS CONTENTS IS THE EXCLUSIVE PROPERTY
OF ECOLYSE, INC. AND LS FURNISHED ON A CONFIDENTIAL BASIS, AND WI1'H THE EXPRESS AGREEMENT THAT 1T WII.L
NEITHER BE USED, SOLD, TRANSFERRED, COPIED, TRACED, PHOTOGRAPHED, NOR REPRODUCED IN ANY MANNER
WHATSOEVER IN WHOLE OR IN PART, NOR ANY ITEM HEREIN BE SOLD, MANUFACTURED OR ASSEMBLED WITfIOUT THE
WRITTIN AGREEMIIVT OF ECOLY9E, INC. THE RECIPIIIVT OF THIS DOCUMENT AGREES NOT TO DISCLOSE TO ANY OTHER
PARTY INFORMATION CONTAINED HEREIN, OR NOT TO USE SUCH INFORMATION, EXCEPT FOR THE SPECIFIC PURPOSE
INTENDED AT THE TIlv1E OF RELEASE OF THIS DOCUMENT.
Document 1litle
BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONITORING SYSTEM
C Revisbn for Costing 2]une 2011 SC
B Revised for Revlew 25 May 2011 SC BL SC
A ISSIIED FOR REVIEW [DRAFf� 22 May 2011 SC BL
Rev 3tad�s Date Originator Checker Approver Cnstomer
Ecolyse Project Na Client and Client Reterence or pocument No. TWaI Pages
Wichita Falls Water Utility — Daniel Nix
11-015 o���t rb. 18
11015-PRO-001-C
Proposal
Ec� lyse, I nc.
M 979-694-6500 • F 979-694-6511 • 11142 Hopes Creek Road • College Station, Texas 77845 • www.ecolyse.com
E cc� lyse I��� Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RAp( FILTRATION AS
MEASURED WITH A PROPOSED MONfTORING SYSTEM
TABLE OF CON'I'ENTS
ABBREVIA'I'IONS ...................................................................................................................... 3
1.0 INTRODUCTION ............................................................................................................. 4
2.0 WORKSCOPE ..................................................................................................................4
2.1 Establlshment and Evaluation of Equivalent Biofilms within Dynamic Flow cells.... 5
2.1.1 Evaluation of Microbial Biofilm Consortium ..................................................................... 6
2.1.2 Establishment of the Equivalent Micmbial Biofilm Consortium ....................................... 6
2.2 Biocide Ef�icacy Against Established Biofilm using Dynamic Flow Cells ................... 6
2.2.1 Biofilm Development in Dynamic Flow cells .................................................................... 6
2.2.2 Prepazation of Sessile Test Cultures .................................................................................... 7
2.2.3 Exposure of Test Coupons with Biofilm to Biocide Solutions ........................................... 7
2.2.4 Prepazation of Biocide ........................................................................................................ 7
2.3 Optimization of Biocide treatment .................................................................................. 8
2.4 P'ilot Test Study Employing Optimized Mitigation and Monitoring Program............ 9
2S Design and Implementation of a Monitoring Program Capable Evaluating
MitigationEfficacy ............................................................................................................ 9
2.6 Analysis of Samples from Dynamic Flow cell Test System ......................................... 10
2.6.1 Enumeration of Surviving Sessile Bacteria ...................................................................... 10
2.6.2 Potentiostat Analysis ......................................................................................................... 10
2.6.3 454 Metagenomics Sequencing .........................................................................................11
2.6.4 Polysaccharide Analysis ..................................................................................................... l l
2.6.5 SEM/EDS and XRD Analysis ............................................................................................11
2.6.6 Personnel Requirements ..................................................................................................... l i
3.0 INVOICING .................................................................................................................... 12
4.0 ESTIMATED COSTS ..................................................................................................... 13
APPENDIX 1: EXPERIENCE BONA FIDAS ...................................................................... 15
APPENDIX 2: EXECUTIVE CONSULTANT CIRCUM VITAE ..................................... lb
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Ec� lyse I��• Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASI�iED WfTH A PROPOSED MONRORING SYSTEM
ABBREVIATIONS
APB Acid-Producing Bacteria
EDS Energy Dispersive Spectroscopy
EPS Exopolysaccharides, the main chemical component of biofilms
GHB General Heterotrophic Bacteria
106 Iron Oxidizing Bacteria
MOB Manganese Oxidizing Bacteria
NACE National Association ot Corrosion Engineers
OCP Open Circuit Potential
PBS Phosphate Buffer Solution
SEM Scanning Electron Microscope
SRB Sulfate-Reducing Bacteria
SS Stainless Steel
v/v Volume to Volume
WFWU Wichita Falis water Utility
XRD X-ray Diffraction
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E c� lyse �� C, • Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONRORING SYSTEM
1.0 INTRODUCTION
Ecolyse, Inc. has been requested to submit a proposal to Wichita Falls water utility (WFWU) to screen,
optimize and test biocides to mitigate MIC within a 316L stainless steel (SS) pilot system. Further to
this, Ecolyse, Inc., has been requested to write and implemeM a monitoring program to measure and
demonstrate the successful mitigation of MIC within the stated system.
2.0 WORKSCOPE
The workscope will be divided into five phases. All biocide evaluations will be performed using system
water found within the fifter system of the utility plant.
A Phase 1 will establish a biofilm in the dynamic flow cell test system that is equivalent to
tfiat found on the 316L SS rack filtration system. Biofilms and tubercles will be evaluated
by metagenomic sequencing to ideMify the viable microbial consortium.
A Phase 2 will evaluate the efficacy of approximately 8 different biocides tested at 2 differ-
ent concentrations {3 different conceMration tests will be costed in the appendix), against
established bacterial biofilms.
O Phase 3 will evaluate the efficacy of the top two biocides (3 different concentrations each)
to prevent biofilm formation and ennoblement of SS coupons.
A Phase 4 wiil evaluate the efficacy and optimize biocide treatrnent to mitigate MIC within
the pilot test systems employing the recommend monitoring program.
A Phase 5 wiil include preparation and implementation of an optimized monitoring program
that will evaluate the efficacy of the biocide to mitigate MIC within the 316L SS rack filtra-
tion system.
A typical biocide evaluation to determine the effective biocide concentration is inadequate to determine
the biocide to mitigate MIC of 316L SS. Therefore, the proposed analysis and testing is designed to
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E cc� l se I n�, • Document #: 11-015-PRO-001-C
y � BIOCIDE SCREENING, OPTIMI7ATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 3161. SS RACK FILTRATION AS
MEASUHED WITH A PROPOSED MONRORING SYSTEM
identify a biocide, concentration, and treatment regime that will mitigate the identified MIC. Following is
a list of the detailed analysis and explanation as to what the data will provide.
Q 454 Metagenomic Genetic Sequencing, an in-situ genetic analysis, will identify both
known and unknown microorganisms with a potential for quantification to determine the
most abundant microorganisms. With ttiis analysis, an accurate determination of the
community structure can be made, as well as the most relevant metabolic activity. This
will assist in formulating appropriate media for bacterial enumeration of viable microor-
ganisms after biocide testing.
n NACE standard TM0194-04 method will be used to enumerate viable microorganisms.
O Polysaccharide analysis will be used to determine the amount of extracellular polymeric
substances (EPS) within solid samples and can be used to estimate e�ent of biofilm for-
mation.
n A potentiostat will be used to follow the shift in open circuit potential (OCP), a phenome-
non called ennoblemeM that precedes pitting in 316L SS. This will allow ennoblement to
be monitored and will assist in determining when an adequate biofilm has been estab-
lished for biocide testing. Enn�lement will be used to evaluate treatment regimes.
A Scanning electron microscopy coupled with energy-dispersive x-ray analysis (EM/EDS)
will be used to evaluate corrosion morphology e.g., pit shape and size and elements
within or associated with the pits.
A X-ray diffraction (XRD) will be used to establish the crystalline structure of solids, specifi-
cally identifying the potential corrosion product.
2.1 Establishment and Evaluation of Equivalent Bioftlms within Dynamic Flow cells
The establishment of a biofilm wRhin the dynamic flow cell that is equivalent to the biofilm on the 316L
SS rack filtration system is essential for proper biocide evaluation and optimizabon. To initiate corrosion,
spec'rfically pitting corrosion on 316L SS, it is necessary to ennoble the SS +300mv. Depositing iron or
manganese oxides on the SS surtace can accomplish the ennoblement of the SS. Microorganisms,
spec'rfica�ly iron oxidizing or manganese oxidizing bacteria, can accomplish the deposition of these ox-
ides on the surtace of the SS. However, Campbell et al., 2003, demonstrated that a consortium biofilm
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E c� lyse I n c• Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASI�{ED WITH A PROPOSED MONITORING SYSTEM
could prevent the ennoblement of SS even with the presence of a manganese oxidizing bacterium L.
disaophora that is known to ennoble SS causing pitting corrosion. Therefore, it is necessary to establ�h
a biofilm in the dynamic flow cells equivalent to the biofilm that is found on the 316L SS racks.
2.1.1 Evaluation of Microbial Biofilm Consortium
Samples will be collected from the 316L SS racks that are near or associated with pitGng comosion.
Samples will be evaluated utilizing 454 metagenomics sequencing to establish the microbial consortium
and to quantify the dominant microbial populations. Data will be evaluated and summarized for com-
parison to test system biofilms.
2.1.2 Establishment of the Equivalent Microbial Biofilm Consortium
A dynamic flow cell system will be set-up and operated at the WFWU so that a continuous supply of
system water is available. The water will be the source of the microbial consortium to establish an equi-
valent biofilm for biocide studies. The dynamic flow cell system will be operated for a period of 28 days
or more to achieve a mature biofilm, capable of causing pitting corrosion on the 316L SS coupons.
Biofilms will be evaluated once a week, utilizing metagenomic sequencing to enumerate and identify the
dominant microorganisms and that information will be correlated with level of ennoblement as measured
by a potentiostat, uRimately identifying how long it takes to establish a mature biofilm capable of causing
pitting corrosion. Metagenomic sequencing data will help establish the basis for a monitoring program
using growth media, the will eritail pertorming a triplicate single serial dilution protocol until dilution to ex-
tinction.
2.2 Biocide Efficacy Against Established Biofilm using Dynamic Flow Cells
2.2.1 Biofilm Development in Dynamic Flow cells
The sessile populations will be evaluated utilizing a dynamic flow cell system instead of the hanging
coupon method described in NACE TM0194-04. The dynamic fbw cell will mimic what occurs in a fbw-
ing system, rather than the stagnant, diffusion-dependent system represented in the NACE standard.
The sessile populations will be developed as biofilms on 0.5 cm 316L SS (biostuds) in a dynamic fbw
cell system inoculated with planktonic microorganisms from the source water. Test coupons containing
a biofilm will be exposed to each biocide solution independently at the recommended concentrations.
Biofilms will be removed by scraping, sonicating, and mixing in a phosphate buffer solution. The num-
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��� ���� ' � � • Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MRIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASI�iED WITH A PROPOSED MONITORING SYSTEM
bers of survivi� sessile IOB, MOB, GHB, APB, and SRB within will be enumerated by the most prob-
able number (MPN) method.
2.2.2 Preparation of Sessile Test Cultures
The dynamic flow cell system will be operated continuously with system water for approximately one
month to establish a mature biofilm as detertnined from ennoblemeM data. After approximately 1 month,
the 316L SS biostuds will be subjected to an established biocide treatment regime. Control test coupons
will be run simultaneously with each biocide. Test coupons will be withdrawn and the numbers of 108,
MOB, GHB, APB, and SRB will be determined by the MPN method.
2.2.3 Exposure of Test Coupons with Biofilm to Biocide Solutions
Immediately following the development of a mature biofilm, the biocide pumps will be turned on and test
water containing the appropriate concentra6on of the biocide will be eluted through the system. A total
of S biocides at 2 concentrations each will be tested. Therefore, a total of 14 biocides will be evaluated.
A single system can test up to 7 biocides with 1 coMrol. Therefore, two separate systems will run simul-
taneously to minimize the time required for a 28iiay test. The biocide(s) concentration(s) as measured
by active component of biocide will be evaluated. Biocides will be evaluated on treatment duration as
well as concentration to be able to compare biocide efficacy direcUy. After the treatrnent period, the bia
cides will be tumed off and onty system water will be eluted through the dynamic flow cell system until
the next biocide treatrnent or the completion of the experiment. The concentration of biocide to be used
in this study will be recommended by WFWU or as determined from manufacturer recommendations.
Samples will be collected at 0 min, 2 hrs., 24 hr., 48 hr., 3 days, 7 days, 14 days. A blank coupon will
be inserted in the sample coupon's place and the dynamic flow cell system will be tumed back on.
2.2.4 Preparation of Biocide
Sterile system water buffered to pH 7 will be prepared in 1000 ml bottles. The biocides will be added to
achieve the desired concentration(s) of biocide(s) at concentrations as measured as active concentration
of vN that will be determined at a latter date. Controls will be set up using distilled water in place of bio-
cides.
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Ec� lyse � n`, • Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONRORING SYSfEM
2.3 Optimization of Biocide treatment
Successful management of MIC within the real system will be achieved by successfully managing risk
factors associated with the identified MIC. In most cases, microbial biofilm and microbial activity pose
the most sign'rficant risk for causing MIC. Therefore, successful mitigating biofilm development may
significantly decrease the risk that could be posed to the system. Interestingly enough, part of biofilm
development is a function of microbial activity. Within an operating system, biofilms develop according
to the following equation:
Biofilm Development =(inoculum • growth) -(biocide efficacy' washout)
According to this equation, the extent to which biofilms develop depends on the size of the inoculum
and growth of bacterial cells. The secondary feature of biofllm development is death of bacterial cells
as influenced by biocide efficacy as well as biofilm detachment that leads to washout.
There will be no development of biofilms in the optimization of the biocides chosen for this study. The
system will be sterile when started. The system will then be filled with system water and allowed to run
for 4 hours to condition the system. After 4 hours, the biocide treatments will begin. For this experiment,
the top two biocides will be selected for further analysis. It is estimated that there will be 3 different
treatment regimes for each biocide with 1 control. For the course of 28 days, the following is an example
of a batch treatrnent.
A Dynamic flow cells set up.
A Continuous flow pumps primed and started. The dynamic flow cells string 1, 2, 3, and 4 started
flowing at 3 ml min. with artificial seawater.
A Samples for time 0 hr will be taken 4 hours after the start of the flow and just before biocide
Veatrnent for all dynamic flow cell strings. Biocide treatment for strings 2, 3, and 4, will proceed
at appropriate dosing as described above for a 4-hour duration.
A Samples for time 4 hr will be taken just after biocide treatment for all dynamic flow cell strings.
C! Samples for time 24 hr will be taken on sVings 1, 2, 3, and 4.
A Samples for time 48 hr will be taken on strings 1, 2, 3, and 4.
A Samples for time 3-day will be taken just before biocide treatrnent for all dynamic flow cell strings.
A Biocide treatrnent for strings 2, 3, and 4, will proceed at appropriate dosing as described above
for a 4-hour duration.
A Samples for time 7 day will be taken just before biocide treatment for all dynamic flow cell strings.
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E cc� lyse I n c• Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONRORING SYSTEM
A Day 10, Biocide treatment for strings 2, 3, and 4, will proceed at appropriate dosing as described
above for a 4-hour duration.
O Samples for time 14 day will be taken just before biocide treaVnent for all dynamic flow cell
strings.
A Biocide treatrnent for strings 2, 3, and 4, will proceed at appropriate dosing as described above
for a 4-hour duration.
n Day 17, Biocide treatment for strings 2, 3, and 4, will proceed at appropriate dosing as described
above for a 4-hour duration.
A Day 21, Biocide treatment for strings 2, 3, and 4, will proceed at appropriate dosing as described
above for a 4-hour duration.
A Day 24, Biocide treatment for strings 2, 3, and 4, will proceed at appropriate dosing as described
above for a 4-hour duration.
n Samples for time 27 day will be taken just before biocide treatment for all dynamic flow cells.
2.4 Pilot Test Study Employing Optimized Mitigation and Monitoring Program
The WFWU has a scale level pilot system for the water filtration. After optimization of the biocide
treatment, it is proposed that this pilot system be implemented to optimize the chemical treatment for the
water treatrnent facility. Upon initial meeting and site tour, the pilot system was evaluated as part of the
study. This pilot system can be used for these studies, although, it may require mod'rfications. Essen-
tially, the system can be run indefinitely, however, it is proposed that it be run for 45 days with samples
being taken every 14 days to monitoring the midgation efficacy.
2.5 Design and Implementation of a Monitoring Program Capable Evaluating
Mitigation Efficacy
Under most biocide treatrnent programs, monitoring is only pertormed to demonstrate the ability of the
biocide to kill microorganisms. However, this form of ineasurement does not demonstrate and nor can it
be exVapolated to demonstrate that MIC is being mitigated. Therefore, it is necessary to write a monitor-
ing program that when implemented, will be able to measure the level of MIC mitigation efficacy that is
being achieved as well as highlight what may be done to achieve the expected level of mitigation. A
custom monitoring program designed to establish sampling locations, analysis to be performed, and
steps for optimization is recommended to be completed prior to the pilot study to ensure that the pro-
gram will provide the data required to mitigate corrosion. It is projected that it will take approximately 3—
5 days to write a complete program.
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Ec� lyse I�� • Dceument #: 11-015-PRO-001-C
� BIOCIDE SCFEENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONRORING SYSTEM
2.6 Analysis of Samples from Dynamic Flow cell Test System
2.6.1 Enumeration of Sunriving Sessile Bacteria
Phase 2 Biocide screening:
Samples will be collected at 0 min, 2 hrs., 24 hr., 48 hr., 3 days, 7 days and 14 days.
Phase 3 Biocide TreatmeM Optimization:
Samples will be collected at 0 min, 2 hrs., 24 hr., 48 hr., 3 days, 7 days, 14 days and 28 days.
Viable microorganisms, with a focus on 106, MOB, APB, GHB, and SRB, will be enumerated. The test
coupon will be transferred into a test tube containing 10 mi sterile PBS solution. The tube and coupon
will be subjected to sonication for 1 minute. This releases the biofilm, which is further dispersed into sus-
pension by mixi� with a vortex mixer. The resulting suspension will be suitable for inoculating into a se-
rial dilution series as previously described. It is proposed that the analysis for these studies will begin
with i 6 samples for the biofilm evaluation study, 112 samples for the biocide screening study, 56 sam-
ples for the biocide optimization study, 3 samples for the pilot study. This will be approximately 187
samples taken during the course of all of the experiments.
2.6.2 Potentiostat Analysis
Through the course of all experiments, including the pilot study, it will be necessary to monftor the
electrochemical potential of 316L SS coupons exposed in the laboratory flow-loop. Potential values of
multiple 316L SS coupons will be automatically recorded versus a single silvedsilver chloride reference
electrode as a function of time using a multiplexed voRmeter wfth data logging capability. Data will be
downloaded onto a computer on-site and distributed for analysis. Detection of potential increase (enno-
blement) will be used as one indication of adequate biofilm establishment for biocide testing. In addition,
potendal monitoring will assist in identifying a treatrnent regime that can prevent pitting corrosion. Poten-
tial ennoblement increases the likelihood of pitGng corrosion initiation, uRimately leading to through-wall
penetration and piping failure. Sharp decrease in potential over time is an indication that pitting cono-
sion has been initiated. The use of multiple 316L SS coupons will accommodate the removal of select
coupons from the flow Ioop at specific intervals over the exposure period. Biofilm development will be
correlated to measured potential values as well as visual indications of corrosion.
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Ec� lyse I n�. • Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTI�,
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WffH A PROPOSED MONfTORING SYSTEM
2.6.3 454 Metagenomics Sequencing
The application of genomic sequencing during all studies will be paramount to understanding the
mitigation of MIC. It is proposed that the analysis be performed at 7 and 28 days during the evaluation
of the biofilm study; on day 14 for the biocide screening study; day 28 for the biocide optimization study,
and 3 samples every 14 days in the pilot study. This will be approximately 20 samples taken during the
course of all of the experiments.
2.6.4 Polysaccharide Analysis
The polysaccharide analysis will help determine the amount of EPS that is present on the coupons and
can be a factor influencir� corrosion. Further to this, the amount of EPS is a function of microbial activ-
ity, and microbial activity predominately influences MIC. Therefore, analysis for potysaccharides as a to-
tal percentage of the solids will be essential for understanding the mitigation of corrosion during all stud-
ies. It is proposed that the analysis be pertormed after 7, 14 and 28 days during the evaluation of the
biofilm study; 7 and 14 days for the biocide screening study; 7, 14 and 21 days for the biocide optimiza-
tion study; and 3 samples every 14 days in the pilot study. This will be approximately 46 samples taken
during the course of all of the experiments.
2.6.5 SEM/EDS and XRD Analysis
Understanding the corrosion in relation to the solids on the coupon will provide the information required
to interpret the ennoblement data. In addition to this, identification of microbial deposi�on of solids can
be potentially ident'rfied. It is proposed that the analysis be performed at 14, and 28 days during the ev-
aluatlon of the biofilm study, 28 days for the biocide optimization study, and 1 sample every 14 days in
the pilot study. This will be approximately 20 samples taken during the course of all of the experiments.
2.6.6 Personnel Requirements
As the biocide evaluation will be pertormed on-site at WFWU, Ecolyse Inc. will have to provide a level of
staffing through the course of the experiments. However, WFWU personnel will conduct most of the in-
oculations, incubations, readings, and maintenance. The following personnel and staffing is proposed
with associated hours for onsite charges:
A Field Technician (To Be provided by WFWU) 25 days
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E c� lyse I n�• Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTR4TION AS
MEASURED WITH A PROPOSED MONITORING SYSTEM
A Project Manager/Junior Microbiology Consultant 10 days
A ElecVochemist 3 days
A Senior ChemisUCoROSion Consultant 2 days
A Executive Microbiologist ConsultaM 3 days
3.0 INVOICING
AII prices quoted are exclusive of Tax.
This quotation is valid for a period of 30 days.
Invoicina information.
The trial will be invoiced 5�o up front and then bi-weekly progress payments in a normal invoicing
process, as based on deliverable or project stage
Due to the nature of the work being undertaken, our quotation is based on best estlmate of time
required. However, should the work scope quoted for be modified as a direct result of customer
requirements, we reserve the right to request a price increase.
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E cc� lyse ' �C, • Dceument#: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONITORING SYSTEM
4.0 ESTIMATED COSTS
Itom DesdipUon Unit Cost Quantlty ToUI Cost
S U.3. S U.3.
I. Enum�ratlon of Survivina Sessib Baetaria
Set Up - Dynamic Flowcells
to indude dean up and malntanence by WFWU personell
Biofilm Devebpment Study $ 100.00 2 $ 200.00
Biocide Screening Study $ 100.00 16 $ 1,600.00
Biocide Optimization SWdy $ 100.00 7 $ 700.00
Subtofal - Set-uo �E 1.800.00
Consumables
Media for SRB, APB, 108 and GHB Estimated 16 cases for each $ 200.00 64 $ 12,800.00
Innoculations W be pertortned by WFWU personell $ - 187 $ -
(MPN method requires 2 boxes of inedia for each baderial type per sample)
Diluent 3 30.00 20 $ 600.00
Needles and Syringes 3 80.00 20 $ 1,200.00
Subtofal, Conaumablss �i 14,800.00
ReooAina and Review
Incubation, data reading, end reporting (To be performed by WFWU) E - 2 $ -
Senfa Consulterrt ReporUng and review $ 1,800.00 1 E 1,800.00
Subtohl, RapoRing snd RavNw � S 1,800.00
Subtohl, Enumentlon of Surviving Sasslk Bacteria S 18,200.00
11. Potentlastet Analvsis
(k is neommendsd that this be made as a purchasa) $ - �$ -
Consuldng Charge for Set-up and operedon (Included in Personel charge) �$ -
Data Review, interprefetbn, Repordng $ 1,800.00 1 $ 1,800.00
Subtotal. PotsntlaststAnalysis �S 1.800.00
III. 454 Mahaanomks Swwncina
Semple collection and Anelysis $ 500.00 20 3 10,000.00
Dafa review, genomic functlonal analysis, and repoRing $ 1,800.00 2 $ 3,600.00
Subtofal, 454 Metagenomics Sequencing �E 13,600.00
N Polvsaccarhlde AnaNsis
Semple cdlectlon and Analysis $ 75.00 46 $ 3,450.00
Dafa review, w/w % analysis, and reporting $ 1,200.00 1 $ 1,200.00
Subtotal, Polyaaccuhide Maysis S 4,650.00
V SEMlEDS and XRD Corroslon Anahrsb
(Corroslon analysls and data intxpretatlon wlll be pertormed against NACE 3T799)
Sample collection and Analysis for SEM/EDS $ 650.00 20 $ 13,000.00
XRD Malysis (�II only perfortn on biofilm dev. Md pibt atudy) $ 500.00 . 4 $ 2,000.00
Data review, w/w % analysis, and reportlng $ 1,200.00 1 $ 1,200.00
Subtotal, SEMIEDS and XRD Corrosion Analysls S 76,200.00
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Ec� lyse �� C• Dceument #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PIIAT SYSTEM TESTING
FOR THE MRIGATION OF MIC ON 376L SS RACK FILTRATION AS
W�ASURED WITH A PROPOSED MONITORWG SYSTEM
VI Personnel and Stafflna
(PsnonnN and StaTflng wifl bs �stlmatsd for th� purpose of this
proposal and wlll ony eonstltue charges with penonnel are onsfEe.)
Field Techniclan (to be provided by WFWU) S - 25 S -
Project ManageNMicrobblogiat Consultant $ 1,200.00 10 $ 12,000.00
Electrochemfst $ 1,400.00 3 $ 4,200.00
Senfor ChemistlCorrosbn Consuttant $ 2,000.00 2 S 4,000.00
Execufive consultaM $ 2,200.00 3 a 6,800•00
Subtotal, Personnel and StaMing S 28,800.00
VII Travel and Acwmodatlons
Fbtel (costs estlmated for the purpose of the proposal) $ 150.00 10 $ 1,500.00
Food end Dining (es�meted tor the proposal) $ 100.00 10 E 1,000•00
Mileage (Estimation of 6 roundtrips at 735 miles Wtal) $ 0.58 1470 S 852.60
Airline Ticket If Required (Brenda Liqle - Corrosion ConsulfaM) $ 750.00 1 a 750•00
Airline Ticket if Required (Jason Lee - Electrochemist $ 750.00 1 $ 750.00
8ublotal, TnvN and Aaomodatbns f 4,852.80
VIII Adminlstratlon Charps
5% charge of total cost $ 4,305.13
8ubtofal, Administradon Chuge S 4,305.13
TOTAL: BIOCIDE SCREENINO.OPTIMIZATION. AND PILOT SYBTEM S 90.407.73
TE8TING FOR THE MITIGATION OF MIC ON 318L S8 RACK
FILTRATION AS MEASURED WITH A PROPOSED MONITORING
YS STEM
14
Ecc� lyse � � � • Document #: 11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MRIGATION OF MIC ON 31 BL SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONITORING SYSTEM
APPENDIX 1: EXPERIENCE BONA FIDAS
Scott Campbell, M.Sc., and Angela Johnson, M.Sc., have been pertorming biocide efficacy and biocide
optimization studies for a combined of 15+ years for all of the major petroleum companies, British Petro-
leum (BP), Shell Exploration and Production (SEPCo.), F�ocon Mobil, Southwest Energy, etc. The bio-
cide efficacy studies performed by Scott and Angela are designed to identify biocide treatments that are
specifically applied to mitigate the identified microbiological problem whether it be microbiological influ-
enced corrosion (MIC) or biofilm development that impacis process systems. Following is a list of cli-
ents/p�ojects, both with Scott and Angela's fortner employer as well as their current employer, Ecolyse,
Inc., demonstrating their experience for performing biocide efficacy testing and optimization.
ASSOCIATED PROBLEM FOR # OF BIOCIDES # OF
� CLIENT COST
BIOCIDE APPLICATION EVALUATED $AMPLES
Southwest Energy Mitigation of biofilm dev. and 67 469 $ 250,000
souri
Biocide New Biocide Efficacy and 5 55 $ 27,900
Intemational determination of Conc.
Champion Biocide Opimizaiton for 6 130 $ 47,700
Technol ies Water Treatment
FMC Mitigation of biofilm dev. and 7 Si $ 34,800
souri
FMC B���de Opimizaiton for 2 g8 $ 39,800
Water Treatment
Design and Establish biocide
Kemira program for Petroleum Consultancy/Ongoing NA $ 200,000
Indus
Lonza Biocide mitigation of MIC 7 35 $17,600
MSA for Biocide
Lonza optimizaiton, formulation, Consultancy/Ongoing NA $ 250,000
field testin
Schlumberger Field evaluation of biocide � 40 $ 47,000
efficac
Shell �Ptimization of biocide for 6 70 $ 65,000
souri and MIC
Evaluation of new biocide for
SEPCo. mitigation of MIC and 3 65 $ 55,100
Sourin
15
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y � BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONITORING SYSTEM
APPENDIX 2: EXECUTIVE CONSULTANT CIRCUM VITAE
SCOiT C. CAMPBELL, M.Sc.
SUMMARY OF QUALIFICATIONS
• Executive Consultant for Oilfield operations — drilling, completians, fracturing, water injection, reservoir
souring, production, and pipelines
• Lead Project Manager for design and implementation of corrosion mitigation and monitoring program for
Intemational dients
• Lead Instructor for National Association of Corrosion Engineers (NACE) Intemal Corrosion of Pipeline
course (Basic)
• Instructor for NACE Internal Corrosion of Pipeline course (advanced)
• Designed and implemented dynamic flow cells for more accurate and representative 6iocide efficacy test-
ing and evaluations for the oil industry to include MIC mitigation.
• Consultation experience to employ through and complete data analysis
• Designed and implemented optimized monitoring programs for platforms and onshore production for mi-
crobial control
• Optimized treatment systems for mitigation of microbial activity in drilling and fracturing systems
• Extensive education in microbial theory, molecular biology, and laboretory procedures
PROFESSIONAL AFFILIATIONS
• Member Society Petroleum Engineers
• SPE Vice Chair for MIC Meeting — Calgary (2008)
• Member NACE
• NACE Chair for TG 214 for Writing document for "State of the Art understanding for Microbial Influenced
Corrosion (MIG)
• NACE Chair of MIC Symposium (2011)
• NACE Vice-Chair of MIC symposium (2010 & 2009)
• NACE Vice-Chair for TG 214 - Re-write of NACE TM0194-94
• NACE Intemal Corrosion Lead Inrtructor
• NACE Committee member MIC corrosion subcommittee
• NACE committee member of eduation sub-committee)
EDUCATION
1997-2002 Montana State University - Center for Biofilm Engineering Bozeman, Mantana
Masters in Microbiology
• Advanced Genetics
• Microbial Ecology
• Microbial Physiology
• Environmental Microbiology - Environmental Engineering - Supporting Area
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� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 316L SS RACK FILTRATION AS
MEASLR�ED WfTH A PROPOSED MONRORING SYSTEM
• Microbial processes atinterfaces
• Chemical Sensors
• Reactions and Reactors
• Procedures
EMPLOYMENT
2009 - present Ecolyse, Inc. College Station, Texas
Vice President Petroleum Services/Exewtive Consultant
• Developed Ecolyse services market for consulting, training, analyticai, chemical evaluations, and field sur-
veys to support retained clientele
• Integrated laboratory protawls to support phage testing on bench scale systems for industry optimization
and applications
• Wrote and implemented training courses for Petroleum Microbiology courses
• Retained as consultant for major chemical manufacture and petroleum production companies
2003 - 2009 Intertek Commercial Microbiology Houston, Texas
Exewtive Consultant/General Manager
• Successfully managed US division office (Commercial Microbiology, Inc.) achieving a 340% growth in 5
yean, from $250,000 to $1.7 million prior to merger with Intertek.
• Consulted for all of the major oilfield operatars for microbial control in drilling, completions, frecturing,
hydrotesting, storage, water injection, reservoir souring, production (water and oil), and pipelines
• Optimized biocide and monitoring programs through operation design for platforms and onshore produc-
tion systems
• Assisted in developing microbial influenced wrrosion (MIC) model that was applied for optimizing biocide,
operetion, and monitoring programs
• Developed Iron bacteria media for the oil industry
• Developed dynamic flowcell for biocide testing
• OFfshore experience wllecting and testing samples
• Analysis of drilling muds, hydrotest waters, pigging trash, fuels and various samples for appropriate bacte-
ria
1997-2003 Centerfor Biofilm Engineering Montana State University
Graduate Student — Microbiology
• Performed chemical analysis using titrations and HPLC
• Developed and executed a successful procedure for in-situ hybridization
• Perfartned ELISA, Northem and Southern analysis, immunaprecipitations
• Performed cryosectioning at SOmm and less on biofilms and cell cuitures
• Isolated and maintained laboretory stock cultures
• Constructed oxygen microelectrodes for chemical analysis in-situ
• Performed extensive phatomicroscopy using canfocal and fluorescent microswpes
1�
E �4J�yJ� In�,• Dceument#:11-015-PRO-001-C
� BIOCIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION � MIC ON 318L SS RACK Fl�TRATION AS
MEASURED WITH A PROPOSED MONRORING SYSTEM
• Teaching assistant for immunology and medical bacteriology
1994-1997 Dr. Gordon McFeters Laboratory Montana State University
• Laboratory Assistant
• Developed and executed a successful protocol for culturing microorganisms on alternative nutrient source
• Performed chemical quantitation of cyanide using HP spectrophatometer and titrations
• Performed gold plating and extraction for sample analysis on ICP-AES
• Perfarmed several types of microbial plating procedures and maintained stock culture
P U BLICATIONS/LECTU RES
Campbell, S., Duggleby, A, Johnson, A, 2011. Conventional Application of Biocides May Lead to Bacterial Cell Injury
Ratherthan bacterial kill Within a Biofilm. NACE corrosion 2011, March 2011, Houston, TX, USA
Willmon, J., Campbell, S., and 55 Commissioning Team, THPS Degradation in the Long-Term Preservation of Sub-sea
Flow-lines and Risers, Submitted for review, NACE Corrosion 2010, March 2030, San Antonio, USA (2030j.
Maxwell, 5. and Campbell, S. 2008. Predicting, Controlling and Monitoring MIC in Seawater Injedion Systems. Pa-
per #283, Recife/PE Brezil. ABRACO Intercorr 2008 2 International Conference
Maxwell, S. and Campbell, 5. 2007. Monitoring the Mitigation of MIC Risk in Pipelines. Calgary Canada, SPE Micro-
biological Influence Corrosion Invitational Conference, 2007
Campbell, 5. 2007. Techniques for Monitoring Microorganisms, specifically SRB, in the Oilfields, Houston, TX. NACE
Technology Week
Maxwell, 5. and Campbell, S. Monitoring the Mitigation of MIC Risk in Pipelines. Paper 06662. NACE Corrosion
2006, March 2006, San Diego, USA (2006).
Kane, R. and Campbell, S. 2004. Real-time corrosion monitoring of steel influenced by microbial activity (SRB) in
simulated seawater injection environments. Paper t104579. Houston, TX. NACE Intemational Conference. Paper.
Campbell, S. Measuri�g the spatial distribution of L discophoro in a 3-species cansortium biofilm using fluorescent
in-situ hybridization. 2002. Technical Advisory Committee Meeting. Bozeman, MT.
Campbell, S., Geesey, G., Lewandowski, Z.,lackson, G. 2002. Influence of the Distribution of the Manganese Oxidiz-
ing BaQerium, Leptothrix discophora, on Ennoblement of 316L Stainless Steel. Paper# 03566. Hauston, TX. NACE
International Conference. Presentation.
Campbell, S., Lewandowski, Z., Geesey, G., Jackson, G. 2002. Influence of the Distribution of the Manganese Oxidiz-
ing Bacterium, Leptothrix discophoro, on Ennoblement of 316L Stainless Steel. Submitted for Review. In Corrosion
Joumal.
Campbell, 5. Influence of the Distribution of the Manganese Oxidizing Bacteriurr�, Leptothrix discophoro, on Enno-
blement of 316L Stainless Steel. Thesis
Ig
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� 810CIDE SCREENING, OPTIMIZATION, AND PILOT SYSTEM TESTING
FOR THE MITIGATION OF MIC ON 31 BL SS RACK FILTRATION AS
MEASURED WITH A PROPOSED MONRORING SYSTEM
Campbell, S., Olson, G., Clark, T., and McFeters, G. 2001. Biogenic production of cyanide and its application to gold
recovery. Joumal of Industrial Microbiology & Biotechnology. 26:134-139.
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