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Report of the Land-Based Freshwater 

Testing of the Siemens SiCURETM 

Ballast Water Management System 

March 15, 2010 

 

Principal Investigator: 

Allegra Cangelosi, NEMWI 

Research Team: 

Lisa Allinger, NRRI, UMD 

Mary Balcer, PhD, LSRI, UWS 

Nicole Mays, NEMWI 

Tom Markee, LSRI, UWS 

Christine Polkinghorne, LSRI, UWS 

Kelsey Prihoda, LSRI, UWS 

Euan Reavie, PhD, NRRI, UMD 

Donald Reid, Consultant 

Heidi Saillard, LSRI, UWS 

Tyler Schwerdt, AMI Engineering 

Heidi Schaefer, LSRI, UWS 

Matthew TenEyck, LSRI, UWS 
GSI/LB/F/A/1 

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Report of the Land-Based Freshwater Testing 

by the Great Ships Initiative 

of the Siemens SiCURETM Ballast Water Management 

System for Type Approval According to Regulation 

D-2 and the Relevant IMO Guidelines 

March 15, 2010 

Allegra Cangelosi, Principal Investigator 

Great Ships Initiative 

Northeast-Midwest Institute 

50 F St. NW, Suite 950 

Washington, DC 20001 

Phone: 202-464-4007 

Email: acangelo@nemw.org 

Approved for Release: 

___________________________ 

PI Signature 

March 15, 2010. 
GSI/LB/F/A/1 

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EXECUTIVE SUMMARY 

The Great Ships Initiative (GSI) provides independent no-cost performance verification testing 

services to developers of ballast treatment systems and processes at a purpose-built, land-based 

ballast treatment test facility located in the Duluth-Superior Harbor of Lake Superior. GSI test 

protocols are consistent with the requirements of the International Convention for the Control 

and Management of Ships Ballast Water and Sediments (International Maritime Organization, 

2004). GSI procedures, methods materials and findings are publicly accessible on the GSI 

website (www.greatshipsinitiave.org). 

In August through October 2009, the GSI conducted land-based tests on the SiCURETM Ballast 

Water Management System in cooperation with German Bundesamt für Seeschifffahrt und 

Hydrographie (BSH), i.e., the German Federal Maritime and Hydrographic Agency. During the 

series of five consecutive valid trials, the SiCURETM Ballast Water Management System was 

evaluated for its ability to: (a) successfully treat ballast water without interruption, (b) meet IMO 

D-2 discharge standards after a five-day holding time, and (c) discharge water after the five day 

retention period that is environmentally benign (i.e., no residual toxicity) pursuant to United 

States Environmental Protection Agency water quality criteria. It should be noted that because 

freshwater zooplankton are in general smaller than their salt and brackish water counterparts, the 

larger regulated size category (greater than 50 μm in minimum dimension) did not incorporate 

all live zooplankton that were present in the source water assemblage. 

The Siemens SiCURETM Ballast Water Management System functioned properly during the five 

consecutive trials, and was highly effective at reducing live organism densities in the fresh water 

ambient conditions of Duluth-Superior Harbor, as amended in these tests to achieve IMOconsistent challenge conditions. Live organisms in the regulated size classes were discharged in 

densities below the IMO D-2 standard. Microbial analyses showed system performance in 

keeping with IMO requirements for bacteria. Chemistry data generated across trials indicated 

the post-retention discharge to have well less than 0.1 mg/L total residual chlorine (TRC) under 

ambient conditions. Ambient water collected immediately after treatment and held in a cold 

environment had TRC and total residual oxidant (TRO) levels which slightly exceeded this level. 

However, in a real world application, the intake water would also be cold, and developers claim 

that the test system is designed to respond to this circumstance (reflected in oxidation-reduction 

potential, or ORP) with a reduction in chlorine generated and injected into the intake stream.

There were no acute toxic effects of treated discharge on any test species across assays and trials. 

Chronic toxicity effects in 100 % effluent were detected in one out of two trials for test species 

of zooplankton and phytoplankton. There were no chronic toxicity effects across organisms and 

trials in 50 % or lower effluent dilutions.
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ACKNOWLEDGEMENTS 

The authors would like to express our sincere gratitude to the Great Ships Initiative (GSI) 

Advisory Committee which provides invaluable input to the GSI Principal Investigator. We also 

wish to thank the ten United States and Canadian Great Lakes Ports which launched the GSI, and 

the Great Lakes Protection Fund which supported the initial scoping exercise. We sincerely 

thank the United States Department of Transportation, Maritime Administration, and National 

Oceanic and Atmospheric Administration for their substantial financial and in-kind support for 

the construction of the state-of-the-art GSI land-based testing facility. We thank the United 

States and Canadian St. Lawrence Seaway organizations, the Legislative Citizens Commission 

on Minnesota Resources, the University of Wisconsin-Superior, and the City of Superior for 

their active financial and/or in-kind support for GSI operations. We are grateful to the German 

Bundesamt für Seeschifffahrt und Hydrographie, BSH, (the German Federal Maritime and 

Hydrographic Agency), for visiting the GSI land-based facility and agreeing to accept data from 

the GSI in support of its type approval determinations, and the Maritime Environmental 

Resource Center for active collaboration as we pioneer ballast treatment evaluations in the 

United States. 
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CONTENTS 

Executive Summary 3 

Acknowledgements 4 

Contents 5 

1.0. Introduction 7 

1.1. The Great Ships Initiative 7 

1.2. The GSI Land-Based RDTE Test Facility 8 

1.3. The Siemens SiCURETM Ballast Water Management System 11 

1.4. Treatment Performance Requirements in Regulation D-2 11 

1.5. Relationship of GSI Testing to G8 and G9 Requirements in IMO 

Convention 12 

2.0. Methods 14 

2.1. Experimental Goals and Design 14 

 2.1.1. Treatment System and Test Facility Calibration Trials 14 

 2.1.2. Valid Trial Series 15 

 2.1.3. Challenge Conditions 16 

 2.2. Water Chemistry Analysis 17 

 2.2.1. Total Residual Chlorine (TRC) and Total Residual Oxidant (TRO) 

 Sample Collection 17 

 2.2.2. Total Residual Chlorine (TRC) Determination 18 

 2.2.3. Total Residual Oxidant (TRO) Determination 18 

 2.2.4. Selected Disinfection By-Products (DBPs) Determination 19 

2.3. Water Quality Analysis 19 

 2.3.1. Analysis of Total Suspended Solids (TSS) 19 

 2.3.2. Analysis of Non-Purgeable Organic Carbon (NPOC) and 

 Dissolved Organic Carbon (DOC), and Determination of 

 Particulate Organic Carbon (POC) Concentrations 20 

2.4. Viable Organism Analysis 20 

 2.4.1. Organisms Greater than 50 μm in Minimum Dimension 21 

 2.4.1.1. Sample Collection 21 

 2.4.1.2. Live/Dead and Size Analysis 21 

 2.4.2. Organisms 10 – 50 μm in Minimum Dimension 22 

 2.4.2.1. Sample Collection 22 

 2.4.2.2. Sample Analysis 23 

 2.4.3. Bacteria 23 

 2.4.3.1. Sample Collection 24 

 2.4.3.2. Sample Analysis 24 

2.5. Ambient Physical/Chemical Water Conditions Analysis 25 
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 2.6. Whole Effluent Toxicity Analysis 25

 2.6.1. Standard Whole Effluent Toxicity Tests 25 

 2.6.2. Cold Whole Effluent Toxicity Tests 26 

 2.6.3. Determination of Total Residual Chlorine (TRC) and Total 

 Residual Oxidant (TRO) Levels for WET Analysis 26 

 2.6.4. Statistical Analysis for WET Assay 26 

 2.6.5. Determination of Quality of Test Organisms for WET Assay 27 

2.7. Data Management 27 

 2.7.1. Data Recording 27 

 2.7.2. Data Processing and Storage 28 

 2.8. Quality Assurance/Quality Control 28 

 2.8.1. Quality Assurance/Quality Control Activities 29 

 2.8.2. Standard Operating Procedures (SOPs) 29 

 2.8.3. Documents and Records 29 

 2.8.3.1. Notebooks 29 

 2.8.3.2. Safety, Operation and Maintenance Records 30

3.0. Results 30 

 3.1. Challenge Conditions 30 

 3.1.1. Physical/Chemical Challenge Conditions 30 

 3.1.2. Biological Challenge Conditions 31

3.2. Viable Organisms in Treated Discharge 33 

 3.2.1. Regulated Plankton 33 

 3.2.2. Regulated Microbes 34 

3.3. Physical/Chemical Water Conditions 35 

3.4. Residual Toxicity 37 

 3.4.1. TRC and TRO Concentrations in WET Tests 37 

 3.4.2. Whole Effluent Toxic Effects 39 

 3.4.2.1. Standard WET Results 39 

 3.4.2.2. Cold WET Results 39 

 3.4.3. Disinfection By-Products (DBPs) 42 

4.0. Discussion of Results 44 

5.0. Conclusions 45 

References 45 

Appendix 1. GSI Land-Based RDTE Facility: Technical Information 46 

Appendix 2. GSI SOPs Relevant to SiCURETM Tests 49 

Appendix 3. Species List of Phytoplankton and Zooplankton Present 51 

Appendix 4. Data Quality Objectives, Criteria and Results 52 

Appendix 5. Operational Data Summary 58 
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1.0. INTRODUCTION 

1.1. The Great Ships Initiative 

The Great Ships Initiative (GSI)1 is regional effort devoted to ending the problem of shipmediated invasive species in the Great Lakes-St. Lawrence Seaway System and globally. In 

support of that goal, the GSI established superlative freshwater ballast treatment evaluation 

capabilities at three scales—bench, land-based, and on board ship. 

The GSI awards its independent status-testing services to developers of ballast treatment systems 

and processes determined to be promising. GSI status-testing is performed at the scale 

appropriate to the state of development of the target treatment system, with the goal of 

facilitating the rapid progression of meritorious ballast treatment systems through the research 

and development and approval processes to a market-ready condition. 

GSI has no involvement, intellectual or financial, in the mechanics, design or market success of 

the actual treatment systems it tests. To ensure that GSI tests are uncompromised by any real or 

perceived individual or team bias relative to test outcomes, GSI test activities are subject to 

rigorous quality assurance, quality control (QA/QC) procedures and documentation. This 

QA/QC attention also assures high quality and credible evaluation findings. 

GSI has worked to standardize and intercalibrate its protocols to evaluate the performance of 

ballast water treatment systems with IMO guidelines, United States Environmental Protection 

Agency Environmental Technology Verification draft protocols, and other test facilities. GSI test 

protocols are as consistent with the requirements of the International Maritime Organization’s 

(IMO) Convention for the Control and Management of Ships Ballast Water and Sediments 

(IMO, 2004) and federal requirements as practicable. In particular, GSI testing directly supports 

IMO G8 and G9 evaluations. GSI procedures, methods materials and findings are also not 

proprietary, and are publicly accessible on the GSI website (www.greatshipsinitiave.org). 

Ms. Allegra Cangelosi of the Northeast-Midwest Institute is the Principal Investigator and 

Manager of the GSI. Researchers from the University of Wisconsin-Superior's Lake Superior 

Research Institute (LSRI), and the University of Minnesota-Duluth's Natural Resources Research 

Institute, among others, provide critical scientific and technical expertise and implementation 

services to GSI's biological research activities, and the GSI generally. Dr. Mary Balcer is the 

project’s lead zooplankton ecologist. Dr. Euan Reavie leads all phytoplankton analysis. Mr. 

Matthew TenEyck, leads the bench-testing and Whole Effluent Toxicity (WET) tests. AMI 

Consulting Engineers provide engineering expertise in support of GSI testing activities. A GSI 

Advisory Committee comprising top-level officials of key stakeholder groups helps steer the GSI 

providing crucial assistance in making GSI award decisions and fund-raising. The GSI Advisory 

Committee includes elected leadership, environmental organizations, port directors and federal 

officials from the United States and Canada, and industry representatives. The American Great 

Lakes Ports Association advises the project, assuring that the GSI is meeting the needs of the 

maritime industry; and coordinating maritime industry and supply chain outreach. 

 

1 www.greatshipsinitiative.org 
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To date, all GSI tests are supported by general project funds which derive from federal, state and 

port grants and contributions, and in-kind contributions by industry, local government and 

universities. Over time, GSI will begin to charge treatment developers for a portion of the testing 

costs associated with type approval testing for United States regulatory purposes. The largest 

contributor of GSI operating funds is the United States Department of Transportation, including 

its Maritime Administration, and the Saint Lawrence Seaway Development Organization. GSI 

also receives significant funds and in-kind contributions from the National Oceanic and 

Atmospheric Administration, the Canadian St. Lawrence Seaway Management Corporation, the 

City of Superior, Wisconsin, and approximately ten U.S. and Canadian ports in the Great Lakes. 

In September and October 2009, the GSI conducted land-based tests on the SiCURETM Ballast 

Water Management System in cooperation with German Bundesamt für Seeschifffahrt und 

Hydrographie (BSH), i.e., the German Federal Maritime and Hydrographic Agency. During the 

series of five consecutive valid trials, the SiCURETM Ballast Water Management System was 

evaluated for its ability to: (a) successfully treat ballast water without interruption, (b) meet IMO 

D-2 discharge standards after a five-day holding time, and (c) discharge water after the five day 

retention period that is environmentally benign (i.e., no residual toxicity) pursuant to United 

States Environmental Protection Agency water quality criteria. 

1.2. The GSI Land-Based RDTE Test Facility 

GSI tests reported here evaluated the biological efficacy of a ballast water treatment system at its 

purpose-built, land-based ballast treatment test facility located in the Duluth-Superior Harbor of 

Lake Superior (figure 1). The facility draws raw intake water from Duluth-Superior Harbor at 

400 m3

/hr to 680 m3/hr. This main flow of intake water can be amended with TSS and endemic 

Harbor algae just prior to entering the facility. 
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Figure 1. The GSI’s Land-Based RDTE Facility in Superior, Wisconsin. 

A Y-split in the intake piping at the facility simultaneously channels one half of the flow (200 

m

3

/hr to 340 m3/hr) to a treatment track and the other half (also 200 m3/hr to 340 m3/hr) to a 

matched control track. The treatment track directs water through the experimental treatment 

system and into a 200 m3

 cylindrical retention tank. The control track by-passes the treatment 

system and channels water directly into a matched control retention tank. After storage, water is 

discharged sequentially from the treatment and control retention tanks at 340 m3/hr to the harbor, 

or to a second set of facility retention tanks where it is then transferred to a wastewater treatment 

facility. A detailed design of the facility can be found in Appendix 1 and information on the 

facility’s validation can be found at http://www.nemw.org/GSI/documents.htm. 

Water is sampled continuously throughout ballasting functions (intake or discharge) through inline sample points. Each sample point is made up of one to three identical sample ports with a 

center-located elbow-shaped pitot tube (90 o

) bent towards the direction of water flow used to 

sample the water. This pitot design is based on a design developed and validated by the U.S. 

Naval Research Laboratory in Key West, Florida, and empirically at GSI. Intake sampling uses 

sample ports at paired intake sample points (SP#2 and SP#3) on the control and treatment tracks 

(figure 2). Discharge sampling occurs through sample ports at sampling points SP#9 or SP#10 

(figure 2). All four SPs are made up of three sample ports. 
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Figure 2. Simplified Schematic of the GSI Land-Based RDTE Facility Showing Location of Sample 

Points, Injection Points, Retention Tanks, and Treatment and Controls Tracks. 

Sample water at a given sampling point (i.e., intake line of the control track, intake line of the 

treatment track, or the discharge line for the control and treatment tracks) is transferred 

simultaneously and continuously throughout ballasting operations (intake or discharge) from 

replicate sample ports to replicate 3.8 m3 sample collection tubs via clean 3.8 cm ID flexible 

hoses and automated flow-controlled pneumatic diaphragm valves. 

An on-site mobile field laboratory (figure 3a) and stationary structure (figure 3b) provide benchscale facilities to support time sensitive assays associated with the GSI land-based tests, 

including live analysis of phytoplankton and zooplankton. The laboratories are climatecontrolled, and have enough bench space to allow for simultaneous microscopic and analytical 

analysis of samples by multiple analysts. 
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Figure 3a. The GSI Mobile Field Laboratory. Figure 3b. The GSI Stationary Laboratory.

1.3. The Siemens SiCURETM Ballast Water Management System 

Siemens Water Technologies developed the SiCURETM Ballast Water Management System 

based on the maritime industry-proven Chloropac® Electrochlorination system for ships’ cooling 

water piping. This latter system was first developed in the early 1970s and has been operated 

onboard over 2,000 vessels. 

SiCURETM has several unique features designed to provide effective treatment of ballast water 

while minimizing risk to the environment, the ship, and its crew. SiCURETM is based on 

electrolysis of seawater and use of hypochlorite as an Active Substance at a viable, “meet the 

demand” dose. The system is designed to inject only as much Active Substance into ballast water 

as required to achieve the necessary level of disinfection. This approach is aimed at eliminating 

over-chlorination and associated risks of corrosion and generation of disinfection by-products. 

The SiCURETM test system evaluated at GSI in August - October 2009 used 5.5 m3/hr side flow 

that, by passing through an electro-chemical generator, was enriched with about 200 mg/L of 

chlorine to treat 200 m3/hr water flow. The system was designed to produce a maximum of 6 

mg/L of chlorine, however the actual dose level was defined by an oxidation-reduction potential 

of the treated water. This control logic was designed to ensure a low biocide residual in the 

water to prevent any corrosion and environmental issues while meeting the IMO Convention’s 

ballast water management standards. 

1.4. Treatment Performance Requirements in Regulation D-2 

The International Convention for the Control and Management of Ships Ballast Water and 

Sediments was adopted by consensus at a Diplomatic Conference at IMO in London on Friday 

13 February, 2004. Annex D-2 of the Convention relates to ballast water performance standards 

for ships conducting ballast water management, including use of a ballast water treatment system 

to effectively treat the ballast water. The regulation states that ships conducting ballast water 

management shall discharge: 
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• Less than 10 viable organisms per m3 greater than or equal to 50 μm in minimum 

dimension; 

• Less than 10 viable organisms per mL less than 50 μm in minimum dimension and 

greater than or equal to 10 μm in minimum dimension; and 

• Discharge of the indicator microbes shall not exceed the specified concentrations. The 

indicator microbes, as a human health standard, include, but are not be limited to: 

o Toxicogenic Vibrio cholerae (O1 and O139) with less than 1 colony forming unit

(cfu) per 100 mL or less than 1 cfu per 1 gram (wet weight) zooplankton samples; 

o Escherichia coli - less than 250 cfu per 100 mL; 

o Intestinal Enterococci - less than 100 cfu per 100 mL. 

1.5. Relationship of GSI Testing to G8 and G9 Requirements in IMO 

Convention 

The fundamental approach of GSI is to conduct independent, scientifically-sound, rigorous, and 

quality assured evaluations of ballast water treatment systems. At the same time, GSI seeks 

immediate relevance of its freshwater land-based testing to regulatory processes such as those 

outlined in the IMO Convention and those under development domestically in the United States 

and Canada. To that end, GSI protocols are rooted in the essential features of the IMO G8 

guidelines for testing, and the draft Environmental Technology Verification protocols under 

development by the United States Coast Guard and United States Environmental Protection 

Agency. All aspects of the testing infrastructure (e.g. flow rate, retention tank size, sample size, 

sample collection and analysis equipment and data logging) are directly consistent with these 

requirements. It formally partners with the Maryland-based Maritime Environmental Resource 

Center (MERC) to assure that GSI freshwater land-based testing can be directly complemented 

by comparable brackish/salt water testing. 

With respect to physical/chemical and biological characteristics of the intake stream, GSI is 

fortunate in that its feed water source naturally meets many of the IMO G8 requirements for 

intake organism densities and physical/chemical conditions during the testing season (table 1). 

Prior to the tests reported here, GSI did not amend water quality in any way in order to avoid 

associated experimental artifacts. In particular, rapid changes in physical conditions (such as 

salinity or total suspended solids) as ambient organisms are being brought in with ballast water 

may cause significant mortality in ambient species, while spiked non-native species may be 

poorly adapted to ambient conditions and decline rapidly as well. 

However, ambient conditions do fluctuate in all natural systems and the time window for this 

specific evaluation of the SiCURETM ballast water management system was limited. Therefore, 

for the tests reported here, GSI agreed to augment intake water to better assure that initial 

challenge water conditions would meet requirements in the G8 guidelines throughout the trial 

series. During initial filling of control and test retention tanks, fine grade Arizona Test Dust, and 

concentrated algae harvested from the Duluth-Superior Harbor were metered into the intake 

stream before the flow split to the control and treatment tracks, to assure adequate concentrations 

of Total Suspended Solids (TSS) and live phytoplankton in intake and control discharge. Details 
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on these processes are provided below. Target intake levels of these parameters appear in Table 

1. Actual levels during testing of the SiCURETM system are reported in the results section of this

document. 

All current protocols, guidelines and requirements are open to interpretation especially in these 

early stages of implementation, and few if any facilities meet all requirements in the strictest 

sense. Accordingly, it is ultimately up to an Administration to decide if the system meets their 

requirements for Type Approval Certification. GSI, along with its sister facility, MERC, sought 

and received German Administration acceptance of its protocols prior to initiating this set of 

tests. 

 

Table 1. Ranges of Various Physical and Biological Parameters in Ambient Water from DuluthSuperior Harbor (June – September) in Comparison to ETV/U.S. Coast Guard and IMO G8 

Recommended Challenge Conditions. 

†

 Generic Protocol for the Verification of Ballast Water Treatment Technologies: Draft v4 2008, U.S. EPA 

Environmental Technology Verification (ETV) Program under contract to U.S. Coast Guard. 

‡ IMO Guidelines for the Approval of Ballast Water Management Systems (G8), October 2008, Annex 4 Resolution 

MEPC.174(58). 

Parameter 

Proposed ETV/

U.S. Coast 

Guard†

Recommended 

IMO G8‡

Historic Ranges

Duluth/Superior 

Harbor 

Target Values for Amended 

Duluth-Superior Water for 

SiCURETM Tests 

Temperature (oC) 10 – 35 – 9 – 22 9 - 22 

Salinity (psu) 0 – 31 Two salinities, >10 

psu difference 0 – 1 0 - 1 

Total Suspended Solids 

(TSS) (mg/L) > 15 > 50 2 – 21 50 

Particulate Organic 

Carbon (POC) (mg/L) > 1 > 5 0.5 – 2.1 <1 

Dissolved Organic 

Carbon (DOC) (mg/L) > 3 > 5 6 – 22 6 - 22 

Zooplankton 

(> 50 μm/m3) > 10,000 > 100,000 100,000 - 3,000,000 100,000 – 3,000,000 

Phytoplankton 

(10 - 50 μm/mL) > 100 > 1,000 25 – 1,200 > 1,000 

Heterotrophic Bacteria 

(CFU/mL) > 1,000 > 10,000 > 1,000 MPN/mL 1,000 MPN/mL 
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2.0. METHODS 

The GSI land-based evaluation of the SiCURETM Ballast Water Management System was carried 

out in keeping with the methods summarized in the MERC-GSI Test Plan (hereafter referred to 

as Test Plan) submitted to and approved by the German BSH and Siemens Water Technologies 

and available at http://www.nemw.org/GSI/MERC+GSITestPlan.pdf. Some refinements were 

necessitated by circumstance or opportunity. The following sections describe how each 

parameter and variable was sampled/analyzed during the SiCURETM trials at GSI. Additional 

details can be found at www.nemw.org/GSI/SOPS.htm. All SOPs relevant to the SiCURETM

tests, as amended, also are presented Appendix 2. Any deviations from these SOPs during the 

performance of the SiCURETM tests were minor and did not affect data quality. More detail on 

these deviations is available upon request. 

2.1. Experimental Goals and Design 

The SiCURETM performance evaluation involved physical and biological characterization of 

water upon ballasting (uptake/intake of water) and comparisons of organisms in control versus 

treated water immediately following treatment and after a five-day in-tank holding time. 

Biological characterizations supported direct comparison with the IMO D-2 organism categories 

and standards. During a series of five consecutive valid trials, the treatment system was tested 

for its ability to: (a) successfully treat ballast water without interruption, (b) meet IMO D-2 

discharge standards after a five-day holding time, and (c) discharge water after the five day 

retention period that is environmentally benign (i.e., no residual toxicity) pursuant to United 

States Environmental Protection Agency water quality criteria. A valid trial was one in which 

control discharge densities of live organisms were at least 10 times the IMO D-2 standard, 

consistent with IMO G8, and in which the facility operated properly. 

2.1.1 Treatment System and Test Facility Calibration Trials

As provided in the Test Plan, GSI first conducted two calibration test runs of the SiCURETM

system. The calibration runs were undertaken to assure the facility and the treatment system were 

operating properly. During these calibration trials, adjustments to the system were documented 

only for internal reference by the treatment developer. 

Necessary adjustments to the facility during this calibration period are documented here for the 

record. During the two calibration runs, line contamination by zooplankton was suspected which 

could have resulted in overestimates of live organisms in treated discharge. To assure facility 

performance and trial independence during the valid type approval test sequence, a thorough 

cleaning of the pipes leading from the control and treatment retention tanks to the sample 

collection tubs was undertaken well prior to the first discharge operation. This cleansing process 

involved filling the pipes with chlorinated water (4 September 2009), and retaining the 

chlorinated water in the pipes for the retention period. Just prior to Trial A discharge, water was 

drained from the pipes and then the pipes were thoroughly flushed with unchlorinated potable 

water (8 September 2009). After this flush, the discharge valves were closed, the sample 

collection tubs were partially filled with potable water, and a zooplankton sample was collected 
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from each tub and analyzed to assure no further organism contamination. In addition, potable 

water for cleaning was obtained from a tanker truck while the potable water reservoir tank at the 

GSI facility was thoroughly cleaned and chlorinated and checked for residual organisms. 

2.1.2. Valid Trials

Once the two calibration trials were complete, the SiCURETM treatment system was set for BSH 

type approval testing by the treatment developer, and the facility physical parameters were 

finalized and set for testing by the GSI Facility Manager. Any further adjustments to either 

component of the testing (the treatment system or the facility) were carefully noted and subject 

to QA/QC documentation. Five valid trials of the SiCURETM system immediately followed the 

calibration runs. 

Because the SiCURETM tests at the GSI facility began later in the summer than anticipated, there 

was a need to complete the testing in the shortest span of time possible to assure adequate 

densities of organisms. To that end, GSI used its capacity to run concurrent tests to overlap trials 

condensing the period required for five trials to 22 days. Consistent with the Test Plan, treatment 

and control intake operations within a given trial were always simultaneous, and treatment and 

control discharge operations were always sequential. Table 2 shows the schedule of the five valid 

tests, including the sequence of intake and discharge operations. 

Table 2. Timing of Intake and Discharge Operations for the Five Valid Siemens SiCURETM 

Trials at the GSI Land-Based Facility in 2009. 

Trial Treatment 

Timing of Operation

Intake Discharge 

A 

Treatment 

4 September 09:48-10:45 9 September 

12:41-13:44 

Control 15:15-16:08 

B 

Control 

11 September 09:52-10:47 16 September 

08:58-09:53 

Treatment 11:51-12:44 

C 

Treatment 

12 September 09:58-10:56 17 September 

09:03-9:56 

Control 11:56-12:51 

D 

Control 

20 September 09:35-10:32 25 September 

08:47-09:41 

Treatment 11:06-11:59 

E 

Treatment 

21 September 09:38-10:36 26 September 

09:02-09:56 

Control 11:38-12:38 
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To assure that discharging the control tank first in no way resulted in contamination of the 

treated discharge, the following steps were taken: 

• Prior to Trial B, the PVC pipes leading from the intake/discharge line to active 

sample collection tubs were replaced with clean, flexible PVC tubing, which were 

removed, cleaned and inspected prior to each remaining treatment discharge 

operation. 

• During all Test Trials A-E the interior of the retention tanks and sampling equipment 

(sample collection tubs, drain spout hose and nozzle, plankton nets, etc.) were cleaned 

according to GSI/SOP/LB/G/O/3 – Procedure for Cleaning the Retention Tanks and 

Other Equipment at the GSI Land-Based RDTE Facility. 

 

2.1.3. Challenge Conditions

Ambient conditions were employed as the physical/chemical challenge conditions, except that 

consistent with the Test Plan, Fine Test Dust was added to the facility intake to assure TSS levels 

were in keeping with IMO G8 guidelines. The solids injection procedure is detailed in 

GSI/SOP/LB/G/O/5 – Procedure for Injecting Organisms and Solids into the GSI Land-Based 

RDTE Facility. Fine Test Dust (ISO 12103-1, A2; nominal 0-80 μm particle size; Powder 

Technology Incorporated; Burnsville, MN) was pre-weighed at LSRI, and sterilized by baking in 

an oven at 190 °C for one hour. One day prior to the test trial, ambient TSS was measured in the 

Duluth-Superior Harbor. On the day of the test trial, the volume of harbor water to be used in 

the Solids Injection System (SIS) tank was determined in order to augment the intake water to 60 

mg/L TSS, and the SIS tank was filled. The prepared Fine Test Dust was poured into the SIS 

tank slowly to prevent clumping, and the dust was mixed for a minimum of 20 minutes prior to 

the start of the trial. The test dust was injected into the intake water for the entire duration of the 

fill at a constant rate using a peristaltic pump located at Injection Point A (figure 2). 

Biological challenge conditions were ambient as well except that organism densities in the 

smaller of the two plankton size classes (i.e., 10 - 50 μm) were enhanced to assure consistency 

with IMO G8 required thresholds. The solids and phytoplankton injection systems were kept 

separate to reduce the risk of interference. The phytoplankton injection procedure is detailed in 

GSI/SOP/LB/G/O/5 – Procedure for Injecting Organisms and Solids into the GSI Land-Based 

RDTE Facility. One to two days prior to the test trial, phytoplankton entities from the DuluthSuperior Harbor were collected and concentrated using 20 - 50 μm plankton nets. The 

concentrated phytoplankton entities were stored at the GSI Land-Based RDTE Facility in 

holding ponds. Prior to injection, the water containing concentrated phytoplankton was mixed, 

sampled, and analyzed for viable cell density. In addition, a sample of Duluth-Superior Harbor 

water was collected to determine the ambient viable cell density. Based on the density of cells in 

the phytoplankton concentrate and ambient intake water, the volume of spiked concentrate that 

would be needed to achieve a concentration of 1500 cells/mL in intake water was calculated. 

This volume was added to an Organism Pressure Injection System (OPIS) vessel. The OPIS 

vessel was pressurized to 25 psi greater than the system pressure. The phytoplankton concentrate 

was added at a constant rate to the intake water via the pressure differential for the entire 

duration of the intake procedure via Injection Point B (figure 2). 
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A static mixer, installed in the main intake line just after the two metering systems (SIS and 

OPIS) and prior to the main system “Y split” (figure 2), assured that the concentrations of these 

additives was equivalent in the control and treatment tracks of the facility. At the end of the five 

day retention period, and just prior to discharge, the tanks were mixed manually with a small 

section of pipe which was pulled along the bottom of the retention tank for a period of 20 

minutes to assure that live organisms, especially spiked algal particles, that may have settled to 

the bottom of the tank during the retention period were accounted for to the greatest extent 

possible in the discharge water analysis. 

2.2. Water Chemistry Analysis 

2.2.1. Total Residual Chlorine (TRC) and Total Residual Oxidant (TRO) 

Sample Collection 

Samples for total residual chlorine (TRC) and total residual oxidants (TRO) analysis were 

collected during intake, retention and discharge. At intake: 

• A 1 L whole water sample was collected from the control intake collection tub; 

• A 1 L whole water sample was collected from the pre-treatment intake collection tub; 

• Three 1 L whole water samples were collected from the filter backwash line at 

approximately 10, 30, and 50 minutes after the start of the intake procedure; 

• Three 1 L post-treatment whole water samples were collected from a sample port 

directly downstream from the treatment system (SP #16) at approximately 10, 30, and 

50 minutes after the start of the intake procedure; 

• Three 1 L whole water samples were collected from a post-treatment sample point in 

the treatment system complex (for use exclusively by Siemens) at approximately 10, 

30, and 50 minutes after the start of the intake procedure, and 

• A 1 L whole water sample was collected post-treatment from a sample collection tub 

(#6). The post-treatment sample from collection tub #6 indicated how much the 

chlorine concentration had degraded over the approximately 45 minutes required to 

fill the collection tub. 

During tank retention: 

• A 1 L whole water sample was collected from the treatment tank on days 3, 4, and 5. 

During discharge: 

• A 1 L whole water sample was collected from the control discharge sample collection 

tub (#1), and 

• Three 1 L whole water samples were collected from treatment discharge sample 

collection tubs (#’s 4, 5, and 6). 
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2.2.2. Total Residual Chlorine (TRC) Determination 

Samples were analyzed for all forms of chlorine, i.e., free chlorine, hypochlorites, and chlorine 

bound to nitrogenous compounds. Analysis took place within 30 – 120 seconds after collection 

to minimize loss of chlorine due to reaction with oxidizable species in the sample. 

For analysis, 100 mL of sample water were transferred from the sample collection container into 

a 150 mL beaker. 1.0 mL of potassium iodide reagent and 1.0 mL of acetate buffer reagent were 

added to each sample. Analysis was conducted with a Thermo Orion Model 97-70 Residual 

Chlorine Electrode connected to an Orion Model 290A pH/mV/ISE meter. 

A 1000 mg/L iodate stock solution (1000 mg/L as chlorine) was used to prepare analytical 

standards, ranging in concentration from 0.100 to 10.0 mg/L daily. The standards were prepared 

in deionized water by making dilutions of the 1000 mg/L iodate stock. Potassium iodide reagent 

and acetate buffer were added to the iodate containing analytical standards. Iodate or chlorine 

present in the standards or samples oxidizes iodide to iodine in an acidic solution. The iodine 

concentration after the reaction was equal to the iodate or chlorine concentration present before 

the reaction. A calibration curve plotting log of the chlorine concentration (x-axis) versus the mV 

response from the Residual Chlorine Electrode (y-axis) was used to determine total residual 

chlorine concentrations in the samples. 

Quality control sample analysis consisted of analyzing duplicate samples and samples spiked 

with known amounts of chlorine. Approximately 10 % of the samples were analyzed in 

duplicate. Spiked samples were only analyzed for the discharge samples because chlorine in the 

intake samples was reduced very rapidly and led to inaccurate spike recoveries. A TRC reference 

sample (QCI-780, ULTRA Scientific) was analyzed daily to confirm the accuracy of the 

calibration standards and samples. 

2.2.3. Total Residual Oxidant (TRO) Determination

Total Residual Oxidant (TRO) analysis was conducted on the same samples as for TRC analysis 

with the exception of the system backwash samples due to high turbidity of these samples. As for 

TRC, TRO analysis took place within 30 to 120 seconds of sample collection to avoid sample 

degradation. 

For analysis, 10 mL aliquots (or smaller amounts if sample concentration was above the 

concentration of highest calibration standard) of sample water were transferred from the sample 

containers into 30 mL beakers. The contents of a Hach DPD Total Chlorine Reagent packet were 

added to each sample. Between 3 and 6 minutes after the addition of the reagent the absorbance 

of the sample was determined using a Spectronic 20D set at a wavelength of 515 nm. The 

absorbance of an aliquot of each sample with no reagent added was also measured and the 

absorbance value was subtracted from that of the sample containing the reagent. This was to 

correct for the background absorbance of each sample. 

A TRO calibration curve was prepared using standards prepared in deionized water utilizing a 

100 mg/L chlorine stock solution. The standards, ranging in concentration from 0.50 to 3.0 
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mg/L, were analyzed in the same manner as described for the samples. The calibration data was 

not linear to the concentration of the upper standard so the calibration curve used a second order 

polynomial fit. 

Quality control sample analysis consisted of analyzing duplicate samples and samples spiked 

with known amounts of chlorine. Approximately 10 % of the samples were analyzed in 

duplicate. Spiked samples were only analyzed for the discharge samples because chlorine in the 

intake samples was reduced very rapidly. A TRC reference sample (QCI-780, ULTRA 

Scientific) was analyzed daily to confirm the accuracy of the calibration standards and samples. 

2.2.4. Selected Disinfection By-Products (DBPs) Determination 

Samples were collected during intake and discharge for analysis of selected disinfection byproducts (DBPs). On intake: 

• Two 1 L whole water samples were collected from the filter backwash line, and 

• Two 1 L samples were collected from the post-treatment sample collection tub (#6). 

On discharge:

• Two 1 L whole water samples were collected from the control sample collection tub 

(#1), and 

• Two 1 L whole water samples were collected from the treatment sample collection 

tub (#6). 

Upon collection samples were stored in coolers with ice packs and transferred to the appropriate 

sample container by the GSI Senior Chemist. The samples were preserved appropriately, based 

on the analysis method, and were shipped cold to Analytical Laboratory Services in Middletown, 

Pennsylvania for analysis of DBPs that might likely be formed through the electrolytic 

chlorination process. The following analyses were conducted: trihalomethanes (THMs), 

haloacetic acids (HAAs), tribromoacetic acids (TBAs), and haloacetonitriles (HANs). In 

addition, sodium chlorate, sodium bromade, and sodium were analyzed

2.3. Water Quality Analysis 

2.3.1. Analysis of Total Suspended Solids (TSS) 

Samples for TSS analysis were collected during intake only as follows: 

• Three 1 L whole water samples were collected from the pre-treatment line (SP #3) at 

approximately 10, 30, and 50 minutes after the start of the intake procedure, and 

• Three 1 L whole water samples were collected from the post-treatment line at 

approximately 10, 30, and 50 minutes after the start of the intake procedure (SP #16). 
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Samples were collected in-line rather than from the sample collection tubs to avoid settling of 

suspended solids. This approach assured a more accurate measurement of solids and organic 

carbon in the intake water. 

For analysis, the samples were vacuum filtered through pre-washed, dried, and pre-weighed 

Whatman 934-AH glass fiber filters. After the sample was filtered it was dried in an oven and 

brought to constant weight. TSS values were determined based on the weight of particulates on 

the filter and the volume of water filtered. 

Quality control sample analysis consisted of analyzing approximately ten percent of the samples 

in duplicate. A TSS reference standard (QCI, 711, ULTRA Scientific) was analyzed on multiple 

occasions along with TSS samples to confirm the accuracy of the data being generated. 

2.3.2. Analysis of Non-Purgeable Organic Carbon (NPOC) and Dissolved 

Organic Carbon (DOC), and Determination of Particulate Organic 

Carbon (POC) Concentrations 

In these tests, NPOC was measured as a surrogate for total organic carbon (TOC), though it may 

be a slight underestimate of TOC. The analytical instrument used to measure NPOC purges the 

sample with air to remove inorganic carbon before measuring organic carbon levels in the 

sample. Thus, the NPOC analysis does not incorporate any volatile organic carbon which may 

be present in the sample. 

Aliquots of the same samples that were analyzed for TSS were also analyzed for NPOC and 

DOC. Before the TSS analysis was conducted, aliquots of approximately 50 mL of the sample 

were transferred to glass bottles and acidified with hydrochloric acid for NPOC analysis. An 

aliquot of the filtrate from the TSS analysis was transferred to a glass bottle and acidified for 

analysis of DOC. A Shimadzu Total Organic Carbon Analyzer (Model TOC-5050A) was 

employed for analysis of both NPOC and DOC. Concentrations of NPOC and DOC were 

determined based on a calibration curve developed on the instrument using organic carbon 

standards prepared from potassium hydrogen phthalate. Reported particulate organic carbon 

concentrations (POC) were determined as the difference between the NPOC and DOC values for 

a sample. 

Quality control sample analysis consisted of analyzing approximately 10 % of the samples in 

duplicate. A reference standard (#516 Demand, Environmental Resource Associates) was 

analyzed daily to confirm the accuracy of the data being generated. 

2.4. Viable Organism Analysis 

Sample water for analysis of viable organisms was simultaneously collected from replicate 

sample ports into identical 3.8 m3

 collection tubs during each intake or discharge operation. 

Volumes retained varied with the operation (intake versus discharge) and treatment (control 

versus treatment), depending upon anticipated organism concentrations, but were always greater 

than IMO guideline volumes. The water in each collection tub constituted an independent time 

integrated replicate sample of the 200 m3 experimental water mass. 
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2.4.1. Organisms Greater than 50 μm in Minimum Dimension 

2.4.1.1. Sample Collection 

 

During the intake operation, i.e. the filling of the treatment and control 200 m3 retention tanks, 

the following time-integrated sample volumes were collected by continuous flow from the intake 

lines simultaneously: 

• 2 - 4 m3

 from the pre-treatment intake line, 

• 2 - 4 m3 from the control intake line, and 

• 2 - 4 m3 from the immediate post-treatment intake line. 

During discharge: 

• One 2 - 4 m3 time-integrated sample was continuously collected from the control 

discharge, and 

• Two to three replicate time-integrated samples of 2 - 4 m3 each (total volume 4 to 9 

m

3

) were continuously collected from the treatment discharge. 

Flow control valves and system logic assured that sample flow rates were equivalent and 

proportional to intake and discharge flow rates throughout each operation. Immediately after 

filling, the entire sample volumes were drained from the sample collection tubs and concentrated 

through 35 μm (50 μm diagonal dimension) plankton nets into 1 L cod-ends for microscopic 

examination. See GSI/SOP/LB/RA/SC/6 - Procedure for Zooplankton Sample Collection. 

2.4.1.2. Live/Dead and Size Analysis 

Live/dead analysis took place within two hours of collecting and concentrating the individual 

samples. Microzooplankton (e.g., rotifers, copepod nauplii, veligers, etc.) and macrozooplankton 

(e.g., crustanceans), all generally greater than 50 μm in minimum dimension (with the exception 

noted below) were analyzed simultaneously by separate taxonomists. Microzooplankton 

subsamples were analyzed in a Sedgewick Rafter counting chamber by examination under a 

compound microscope at a magnification of 40X to 100X. Macrozooplankton were analyzed in 

a Ward’s Counting Wheel at a magnification of 20 to 30X using a dissecting microscope. Due to 

high densities, quantification of zooplankton in intake and control discharge samples required 

analysis of sub-samples and extrapolation to the entire sample volume. For these samples, a 

subsample was removed for analysis using a Henson-Stempel pipette. The treatment discharge 

samples had lower densities allowing analysis of a greater proportion of the sample. Treatment 

discharge samples were split in half using a Folsom Plankton Splitter. Half of the sample was 

analyzed for macrozooplankton and the other half was examined for microzooplankton. The 

proportion and total concentration of live versus dead organisms was determined using standard 

movement and response to stimuli techniques. 

Quality assurance measures included live/dead analysis of at least 10 % of the samples by two 

separate taxonomists. The average percent similarity of taxonomic identification (live organisms 
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only) and the average relative percent difference of the number of live organisms counted were 

calculated for all second analyses. These data quality measurements were compared against the 

data quality objectives outlined in the GSI Quality Assurance Project Plan (GSI, 2009), and the 

percentage of data quality measurements meeting the data quality objectives was determined for 

microzooplankton and macrozooplankton separately. 

Because freshwater zooplankton are in general smaller than their salt and brackish water 

counterparts, the larger regulated size category (greater than 50 μm in minimum dimension) does 

not incorporate all live zooplankton that may be present in a freshwater assemblage. This 

freshwater phenomenon raises special issues with respect to assessing zooplankton densities for 

the purpose of comparison with the IMO D-2 standard. It cannot be assumed that every 

zooplankton individual meets the size criterion in the strictest sense. Moreover, the smaller 

species must be analyzed using a compound microscope on a slide under a cover slip as noted 

above. Normally, this approach at times precludes direct sizing of an organism’s minimum 

dimension during live analysis. During these tests, the Lecanid rotifer figured prominently in 

treated discharge samples. The minimum dimension of this saucer-shaped organism (depth) was 

never visible to the microscopist for direct measurement. To accommodate to this situation, 

following live analysis, samples were preserved with 10 % Lugol’s solution and numerous 

Lecanid rotifer specimens were extracted and directly measured in all dimensions. This 

evaluation showed the organisms in this taxonomic group to be generally less than 50 μm in 

actual minimum dimension. Live individuals in the Lecanidae family present in treated discharge 

were therefore not added to the tally of regulated live organisms, but were counted and 

documented as live zooplankton for consideration by the regulatory authority. See 

GSI/SOP/LB/RA/SA/2 - Procedure for Zooplankton Sample Analysis. 

 

2.4.2. Organisms 10 – 50 μm in Minimum Dimension 

2.4.2.1. Sample Collection 

 

For live analysis of organisms 10 – 50 μm in minimum dimension, one sample of 1 L was 

collected immediately after filling from the pre-treatment sample collection tub and one sample 

of 1 L was collected from the immediate post-treatment sample collection tub. During discharge, 

one sample of 1 L was collected from the control tank via sample collection tub, and three 

samples of 1 L each were collected from the replicate treatment sample collection tubs. Analysis 

occurred on-site within 1.5 hours of sample collection, with samples stored in coolers during the 

interim. Prior to analysis, samples were concentrated through a 10 μm plankton net and stored in 

a 25 mL sample container. See GSI/SOP/LB/RA/SC/3 - Procedure for Algae/Small Protozoa 

Sample Collection. 
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2.4.2.2. Sample Analysis 

 

For analysis, a 1.5 mL subsample of the concentrated sample was transferred to a 2 mL sample 

container, with 4 μL of FDA stock solution added. The subsample was then allowed to incubate 

in the dark for 5 minutes. the 1.5 mL incubated algae sample was mixed and 1.1 mL was 

immediately transferred to a Sedgwick-Rafter cell, covered and placed on the stage of a 

microscope that was set for simultaneous observation using brightfield and epifluorescence. At 

least two horizontal transects were counted (an area known to reflect greater than 1 mL of 

original sample water). If time permitted, additional transects were counted to increase statistical 

power. This resulted in greater than 100 live cells counted from the pre-treatment intake and 

control discharge samples, and often fewer than 10 live cells counted in two transects for posttreatment intake and treatment discharge samples. Single cell entities and cells comprising 

colonial and filamentous entities were characterized as follows: alive = cells showing obvious 

green fluorescence from cell contents; dead = cells showing no or very little evidence of green 

fluorescence from cell contents; and ambiguous = cells or entities that cannot be clearly 

identified as alive or dead (should be uncommon). Records were kept of transect lengths and 

widths so that the total counted area and volume analyzed could be calculated later. 

Entities less than 10 μm in all visible dimensions or greater than 50 μm in minimum dimension 

were not counted. Counting and measurement of all other entities followed standard procedures 

for individuals (length and width), colonies (e.g., number of cells, cell length and width) and 

filaments (e.g., number of cells, cell length and width or total filament length if cells could not be

discerned). The remaining concentrated sample in the 25 mL bottle was archived using a 

preservative (formalin or Lugol’s) for long-term storage. 

Quality assurance measures included analysis of at least 10 % of the samples by two separate 

taxonomists using a dual-headed microscope (i.e., both taxonomists analyzed the same sample at 

the same time). The average percent similarity of taxonomic identification and the average 

relative percent difference of the number of live organisms counted were calculated for all 

second analyses. These data quality measurements were compared against the data quality 

objectives outlined in the GSI Quality Assurance Project Plan (GSI, 2009), and the percentage of 

data quality measurements meeting the data quality objectives was determined. See 

GSI/SOP/LB/RA/SA/1 - Procedure for Algae/Small Protozoan Sample Analysis. 

2.4.3. Bacteria 

Control and treatment samples were collected and analyzed for heterotrophic bacteria, two 

specific indicator pathogens: E. coli and enterococci, and viable toxigenic Vibrio cholerae. 
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2.4.3.1. Sample Collection 

One liter whole water samples were collected as follows: 

• On intake, three were collected immediately after filling from the pre-treatment 

sample collection tubs, and three were collected from the post-treatment sample 

collection tubs. 

• On discharge, three were collected from a control sample collection tub and three 

were collected from a treatment sample collection tub. 

All samples were collected according to GSI/SOP/LB/RA/SC/4 – Procedure for Microbial 

Sample Collection, and were immediately transported in an insulated cooler to the LSRI and 

analyzed as individual replicates. 

2.4.3.2. Sample Analysis 

Viable heterotrophic bacteria were enumerated according to GSI/SOP/BS/RA/MA/1 – Procedure 

for Quantifying Heterotrophic Plate Counts (HPCs) using IDEXX’s SimPlate® for HPC Method. 

This method utilizes the IDEXX SimPlate® for HPC Method (IDEXX Laboratories, Inc.; 

Westbrook, Maine), which is based on IDEXX Laboratories’ patented multiple enzyme 

technology. 

The presence and abundance of E. coli (GSI/SOP/BS/RA/MA/4 - Procedure for the Detection and 

Enumeration of Total Coliforms and E. coli Using IDEXX's Colilert®) and enterococci 

(GSI/SOP/BS/RA/MA/3 - Procedure for the Detection and Enumeration of Enterococcus using 

EnterolertTM) were determined using Colilert® and EnterolertTM, respectively, which are both 

based on IDEXX’s patented Defined Substrate Technology (DST®). 

RNA and DNA colony blots were prepared at the LSRI following GSI/SOP/LB/RA/SA/3 - 

Procedure for the Colony Blot Preparation for Enumeration of Culturable Vibrio chloreae, a 

procedure in which the RNA or DNA of potential Vibrio Cholerae, and a limited number of 

additional species which may grow on the selective media, is fixed to a filter. Filters which 

exhibited colony growth were then shipped to the Maryland Pathogen Research Institute at the 

University of Maryland (College Park, MD) for analysis of potential viable toxigenic V. 

cholerae. Viable toxigenic V. cholerae is assayed with a commercial DFA kit specific for 

serogroup O1 (New Horizons Diagnostics) using monoclonal antibodies tagged with fluorescein 

isothiocyanate (FITC) (Hasan et al., 1994). 

Quality control samples included a media blank and a positive control for E. coli/total coliforms 

and Enterococcus spp.; a media and peptone-saline diluent blank for heterotrophic bacteria; and 

a thiosulfate citrate bile salts sucrose (TCBS) agar blank, and DNA, and RNA blanks for Vibrio 

spp. Quality assurance measures included analysis of at least 10 % of the samples in duplicate. 

The average relative percent difference of all duplicates analyzed during the test trials was 

calculated separately for E. coli, Enterococcus spp., heterotrophic bacteria, and Vibrio spp. In 
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addition, at least 10 % of the samples were counted by two separate analysts and the average 

relative percent difference for all second counts was determined. These data quality 

measurements were compared against the data quality objectives outlined in the GSI Quality 

Assurance Project Plan (GSI, 2009), and the percentage of data quality measurements meeting 

the data quality objectives was determined. 

2.5. Ambient Physical/Chemical Water Conditions Analysis 

Temperature, salinity, dissolved oxygen, chlorophyll fluorescence, turbidity and pH were 

measured every 15 minutes during the test trials by two identical multi-parameter probes 

(calibrated according to manufactures specifications) placed, one each, into the control and test 

tanks. A calibrated, hand-held instrument was used to measure temperature, salinity, and 

dissolved oxygen from the control sample collection tub, the pre-treatment sample collection tub, 

and post-treatment sample collection tub during intake. In addition, temperature, salinity, and 

dissolved oxygen were measured during discharge from one control sample collection tub and 

two or three treatment sample collection tubs. See GSI/SOP/LB/RA/SC/8 - Procedure for 

Collecting Physical/Chemical Data and Samples at the GSI Land-Based RDTE Facility.

2.6. Whole Effluent Toxicity Analysis 

GSI’s whole effluent toxicity testing of the SiCURETM system involved tests for chronic toxicity 

involving three freshwater species as arrayed in table 3. Toxicity tests were conducted on treated 

water from all five test trials. 

Table 3. Standard Operating Procedures Relative to Whole Effluent Toxicity Testing. 

2.6.1. Standard Whole Effluent Toxicity Tests 

One set of tests—Standard Whole Effluent Toxicity Tests (Standard WET)—measured toxicity 

following five days storage in the land-based facility’s 200 m3

 retention tanks. For these tests, 

samples were collected for analysis of residual toxicity at discharge. Sample water, stored in 

large HDPE containers, was immediately transported to the LSRI and was used immediately 

upon arrival to set up the Standard WET tests. Following initial set up of the tests, the remaining 

sample water was held at 4 °C in the dark to retain as much of the initial toxicity as possible, and 

GSI SOP Code Test Type Test Species Test Endpoint 

GSI/SOP/BS/RA/WET/1 Chronic Cladoceran 

(Ceriodaphnia dubia)

Survival and 

Reproduction 

GSI/SOP/BS/RA/WET/2 Chronic Fathead Minnow 

(Pimephales promelas) Survival and Growth 

GSI/SOP/BS/RA/WET/3 Chronic Green Alga 

(Selenastrum capricornutum) Growth 
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portions of the discharge sample water was warmed to 25 °C each day to serve as renewal water 

for the bioassay. A dilution series, using Duluth-Superior Harbor water, was run for each 

species. All tests were conducted in temperature-controlled incubators, water baths, or at 

ambient room temperature following the SOPs listed in table 3. 

2.6.2. Cold Whole Effluent Toxicity Tests 

A second set of trials—Cold Whole Effluent Toxicity (Cold WET) tests—was conducted to 

estimate the TRC, TRO and toxicity effects on organisms under cold water conditions. Though 

informative, it should be noted that in these assays the SiCURETM system’s internal logic that 

self-regulates chlorination of intake water based on oxidation-reduction potential (ORP) was not 

engaged resulting in higher chlorine levels in discharge samples than the treatment developer 

believes will occur under actual cold ambient conditions. Treated water was 

collected continuously from a sample port just downstream of the treatment system (SP #15) 

and diverted into a sample collection tub during the filling of the treatment retention tank. A 50 

L whole water subsample was extracted and placed in a dark, refrigerator set at 4 oC for five 

days, thus simulating cold temperature tank retention. A portion of the sample water was 

warmed to 25 °C prior to initial set up of the Cold WET assay, and was warmed prior to daily 

renewal as described above for the Standard WET assay. There was no dilution series used for 

the Cold WET assay; test organisms (Selenastrum capricornutum, Ceriodaphnia dubia, and 

Pimephales promelas) were exposed to 100 % sample water. The Cold WET assay was 

conducted concurrently with the Standard WET assay following the SOPs listed in table 3. 

2.6.3. Determination of Total Residual Chlorine (TRC) and Total Residual 

Oxidant (TRO) Levels for WET Analysis 

Samples for TRC and TRO were collected from the stock exposure solutions daily. Analysis of 

these samples was conducted in the same manner as for the samples from the testing at the landbased facility (see sections 2.2 – 2.3) with the exception that smaller sample volumes (20 mL) 

were used due to the limited volume of sample available. Amounts of reagents added to the 

samples were adjusted so that a constant ratio of reagent to sample volume was used. 

Standards were prepared, analyzed and subjected to quality control in the same manner as 

described for the land-based analysis (see sections 2.2 – 2.3). Spikes were only performed on 

deionized water due to the rapid degradation of chlorine in Duluth-Superior Harbor water. 

2.6.4. Statistical Analysis for WET Assay

Data was analyzed using the Comprehensive Environmental Toxicity Information Systems 

program (version 1.7, Tidepool Scientific Software, McKinleyville, CA). Data analyses included 

normality, homogeneity of variance, one-way analysis of variance (ANOVA), and suite of tests 

for comparison between treatment means. Non-normal survival data was transformed using the 

natural log (EPA, 2002) to normalize the data. The endpoints of the chronic toxicity tests were: 

• Lowest Observed Effect Concentration (LOEC), i.e., the lowest concentration in a 

test with a statistically significant difference in response from the control response. 
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• No Observed Effect Concentration (NOEC), i.e., the highest concentration in a test 

for which there was no statistically significant difference in response from that of the 

control. 

• Median Lethal Concentration (LC50), i.e., the concentration resulting in death of 50 % 

of exposed individuals by a predetermined time. 

• Effective Concentration (EC25), i.e., the concentration resulting in inhibiting a 

biological function (e.g. growth, reproduction) of 25 % of exposed individuals by a 

predetermined time. 

These measures are extrapolations of statistical results to the experimental endpoints. Mean 

percent survival, mean dry weight values, mean cell density, and mean number of young per 

female for the laboratory controls and treatments were analyzed with a statistical significance 

level of 0.05. 

2.6.5. Determination of Quality of Test Organisms for WET Assay

Whole Effluent Toxicity tests were initiated with healthy, vigorous organisms. To determine the 

overall health of the test organisms, reference toxicant tests were performed with Ceriodaphnia 

dubia and Pimephales promelas prior to the start of each definitive test or at least once per 

month. To determine the validity of the Standard and Cold WET tests, percent survival, dry 

weights of survivors, mean cell density, and mean number of young per female in the controls 

were compared to the test acceptability criteria published in the U.S. EPA’s Short-term Methods 

for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms 

(4th edition, 2002). Class I standardized weights were used as a check for the organism drying 

process and the performance of the balance. Daily and weekly calibration of test meters ensures 

optimal performance. 

2.7. Data Management 

2.7.1. Data Recording

Specific forms (i.e., by size class of organism, by scale of testing, etc.) were used to record 

sample collection and analysis data. Specific forms were also used to record sample custody, 

handling and storage information for those samples that were not analyzed on site at the landbased facility or at the LSRI (e.g., disinfection byproducts and Vibrio cholorae DNA and RNA 

colony blots). GSI Senior Personnel were responsible for ensuring that the forms were correctly 

filled out and timely. They were also responsible for maintaining the forms on file, creating 

electronic copies, and posting to the GSI SharePoint intranet website for storage and archiving. 

Technical systems audits (TSAs) were conducted during each intake and discharge operation for 

all test trials. During each TSA the GSI Assistant QA/QC officer observed data recording 

procedures related to sample collection, zooplankton analysis, phytoplankton analysis, and/or 

chemistry analysis and checked for compliance. Problems identified during each TSA were 

documented on the specific Technical Systems Audit Checklist for that trial’s intake or 

discharge. 
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Following the completion of SiCURETM test trials, a thorough review of all data sheets and 

laboratory notebooks was completed to ensure compliance with the documentation procedures 

outlined in all relevant GSI SOPs and in the GSI Quality Assurance Project Plan (QAPP). A 

Technical Systems Audit Checklist was completed during observation of sample collection and 

analysis activities, and during the data review. A QA/QC Log Book was used to document any 

additional data verification and validation activities. The TSA checklist and QA/QC log book 

were scanned to electronic format and posted to the GSI SharePoint website. 

2.7.2. Data Processing and Storage 

All original datasheets were stored in three-ring binders, each with a unique identification code 

specific to the SiCURETM system tests. All log books were also given a unique identification 

code and are specific to SiCURETM. At least one backup copy (i.e., an electronic copy stored on 

the GSI SharePoint website) was made of all completed datasheets, and in some cases additional 

hardcopies were also made. The raw data is in the custody of the appropriate GSI Senior Staff 

Member until the final technical report has been approved, after which time it will be archived 

by the GSI Assistant QA/QC Officer on the University of Wisconsin-Superior campus for a 

period of at least five years. 

A database designed using the Microsoft Access software suite was used to store, manage and 

process phytoplankton and zooplankton data. Microsoft Excel was used in conjunction with the 

database to create various dataset formats for subsequent analysis. Microsoft Excel was also 

used to store, manage, and process microbial, water chemistry, water quality, and whole effluent 

toxicity data. Database entry and maintenance was the responsibility of the GSI database 

management staff. Regular checks for data entry errors were conducted by comparing database 

records and Excel spreadsheets with the original paper data sheets. This was a manual inspection 

process and though rather time consuming, was an essential procedure for discovering errors. 

After examination and QA analysis, the data distribution files from the Access database were 

posted to the LSRI’s Local Area Network (LAN) in an organized hierarchical folder system. A 

backup of the database was also made regularly to avoid any loss of data following 

computer/electronic glitches. Files were also posted to the GSI’s SharePoint website to provide a 

secondary data backup/storage mechanism. 

2.8. Quality Assurance/Quality Control 

The GSI’s Quality Assurance Project Plan (QAPP) outlines the management activities the GSI 

undertakes to assure the credibility of its land-based research activities. The plan covers QA/QC 

data quality indicators, evaluation processes, performance measures and acceptance criteria; 

instrument certification and calibration; personnel training requirements; QA/QC of documents 

and records; data management; and QA/QC assessments and response actions; etc. The plan is 

updated annually, with a specific process used for review, comment, approval, distribution and 

posting. It closely follows the format of the U.S. Environmental Protection Agency’s (EPA’s) 

“EPA Guidance for Quality Assurance Plans”. 
GSI/LB/F/A/1 

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2.8.1. Quality Assurance/Quality Control Activities

The GSI Assistant QA/QC Officer conducted Technical Systems Audits (TSAs) during each 

Trial’s intake and discharge operation, as well as a thorough data verification and validation 

following the completion of SiCURETM tests to ensure that all information was complete, that 

SOPs were correctly followed, and that QA/QC objectives were met. The results of these 

observations were documented on a pre-printed TSA Checklist and in a QA/QC log book 

specific to SiCURETM. The GSI Assistant QAQC Officer created an electronic copy of each TSA 

Checklist and the QA/QC log book and posted those to the GSI SharePoint website. Once the 

final technical report is finalized, the original QA/QC data will be archived on the UWS campus 

for a period of at least five years. 

2.8.2. Standard Operating Procedures (SOPs) 

GSI SOPs were developed by the relevant GSI senior personnel in conjunction with the GSI PI 

and GSI Senior QA/QC Officer. The SOPs follow a common format and all include specific 

QA/QC procedures and metrics. The Senior QA/QC Officer is responsible for distributing the 

SOPs to the relevant parties for approval. Draft and final copies of all SOPs relevant to the 

SiCURETM tests were posted to the GSI SharePoint website; the final version was also posted to 

the GSI public website (www.greatshipsinitiative.org). SOPs were updated on an as-needed 

basis. 

2.8.3. Documents and Records 

The GSI Senior QA/QC Officer is responsible for maintaining all electronic documents and 

records for a period of five years unless custody is transferred using a chain of custody form. 

Electronic versions of all GSI documents and records are saved to the GSI SharePoint website 

once complete. Hard copies of GSI documents and records are scanned and also saved to the GSI 

Sharepoint website. Due care and diligence will be taken to properly dispose of documents and 

records that are no longer required after the five year period has lapsed. Disposal procedures will 

involve electronic deletion of documents and records from the GSI SharePoint website and the 

personal computers of GSI personnel, as well as manual shredding of hard copies. 

2.8.3.1. Notebooks 

Bound field and laboratory notebooks were used to record observations, sampling details and onsite laboratory and field measurements. These notebooks were assigned a unique identification 

code by the GSI Assistant QA/QC Officer, in order to maintain a record of all laboratory 

notebooks specific to the SiCURETM tests. Notebooks were also used to record instrument and 

equipment calibration and maintenance information. All notebooks were examined periodically 

during the Technical Systems Audits by the GSI QA/QC officers and were checked for 

compliance with GSI SOPs following the completion of testing. Problems identified by the 

periodic QA/QC review were documented and included in a training/response file. 
GSI/LB/F/A/1 

March 15, 2010 

Page 30 of 58 

2.8.3.2. Safety, Operation and Maintenance Records 

The GSI Land-Based RDTE Facility Operations Manager, along with safety staff at AMI 

Consulting Engineers (Duluth, MN) provided safety training to all relevant GSI personnel prior 

to the start of the SiCURETM tests. The GSI Land-Based RDTE Facility Operations Manager 

ensured that all forms associated with safety, operation and maintenance (i.e., confined space 

entry permit forms) were correctly filled out and maintained on file. 

3.0. RESULTS 

Five sequential valid tests of the Siemens SiCURETM Ballast Water Management System were 

completed consistent with the Test Plan accepted by the German BSH as meeting IMO G8 and 

G9 guidelines. Standard and Cold WET chemical assays were conducted in conjunction all of 

these trials, and valid Standard and Cold WET biological assays were completed with two to five 

of the these trials, depending upon the test species and assay type.

3.1. Challenge Conditions 

3.1.1. Physical/Chemical Challenge Conditions 

Dissolved organic carbon (DOC) and total suspended solids (TSS) values were in the range or 

well in excess of the IMO recommended minimum. Intake water salinity during these tests was 

always less than 1 PSU. Ambient particulate organic carbon (POC) was consistently below the 

IMO guideline across trials, probably due to the late-season timing of the tests. A summary of 

physical/chemical conditions of intake and discharge water along with the recommended ranges 

in the IMO G8 guidelines appear in table 4. 
GSI/LB/F/A/1 

March 15, 2010 

Page 31 of 58 

Table 4. Average Concentration (± Std. Dev.) of Total Suspended Solids (TSS), Dissolved Organic 

Carbon (DOC), and Particulate Organic Carbon (POC) in Challenge Water During Five Trials of the 

Siemen’s SiCURETM Ballast Water Management System. 

Parameter IMO G8 

Guideline Sample Trial A Trial B Trial C Trial D Trial E 

TSS (mg/L) 

>50 Intake 34.2 ± 9.4 53.1 ± 0.8 54.7 ± 3.2 53.0 ± 2.6 54.4 ± 1.1 

Post-Treatment 36.5 ± 5.9 48.1 ± 1.6 47.0 ± 0.9 49.2 ± 2.7 47.2 ± 0.8 

DOC (mg/L) 

>5 Intake 7.8 ± 0.03 8.4 ± 0.3 8.5 ± 0.2 8.8 ± 0.2 8.9± 0.2 

Post-Treatment 7.7 ± 0.2 8.9 ± 1.0 8.6 ± 0.2 9.1 ± 0.2 8.3 ± 0.1 

POC (mg/L) 

>5 Intake 0.3 ± 0.4 0.0 ± 0.4 0.1 ± 0.2 0.5 ± 0.5 0.6 ± 0.6 

Post-Treatment 0.7 ± 0.5 0.0 ± 1.0 0.2 ± 0.4 0.2 ± 0.4 1.3 ± 0.1 

Salinity (PSU) <3 Intake and 

Discharge <1 <1 <1 <1 <1 

3.1.2 Biological Challenge Conditions 

Densities of live organisms on intake and control discharge were consistent with or exceeded 

IMO G8 guidelines during all five trials. In the greater than 50 μm size class, the intake density 

of live organisms ranged 287,920 to 1,037,822 with an average of 656,824, meeting the required 

density of 100,000 live organisms per m3. The density of live organisms in the control discharge 

samples ranged from 321,058 to 742,741, with an average of 463,141, well exceeding the 

required density of 100 per m3. Intake concentrations of the smaller planktonic organisms (10 - 

50 μm) also were consistently greater than the required 1000 cells/mL (ranging from 1179.6 to 

1832.5 per mL with an average of 1580.1). The control discharge densities of these smaller 

organisms also exceeded the G8 requirement of 100 live cells per mL (with the exception of the 

Trial E for which the value was 95.6) with an average of 150.3. Table 5 summarizes these 

values. 
GSI/LB/F/A/1 

March 15, 2010 

Page 32 of 58 

Table 5. Live Plankton Concentrations in Test Facility Intake Water and in Control 

Discharge Across Trials. 

Live 

Organism 

Size Category

IMO G8 

Guideline Sample Trial A Trial B Trial C Trial D Trial E 

> 50 μm (#/m3)

>100,000 Intake 769,661 627,566 1,037,822 287,920 561,153 

>100 Control 

Discharge 321,058 486,506 330,787 434,611 742,741 

10 – 50 μm 

(#cells/mL) 

>1000 Intake 1179.6 1832.5 1563.1 1683.2 1642.1 

>100 Control 

Discharge 178.8 204.9 136.9 135.4 95.6 

Plankton diversity also met IMO requirements. The zooplankton community at the test site was 

dominated by the rotifers Keratella (loricate) and Polyarthra and Synchaeta (illoricate or softbodied) which comprised over 80 % of total density. Copepod nauplii, dreissenid mussel 

veligers, copepod copepedites, and bosminid cladocerans were also common in the zooplankton 

community. The density of live soft-bodied rotifers decreased considerably over the five day 

holding time of some of the trials, and was extremely low upon discharge. Densities of loricate 

(hard bodied) rotifers decreased little or in some cases increased during the holding period in the 

control water mass. The smaller regulated size class (10 – 50 μm) was dominated by 

phytoplankton entities of diatoms, green algae, blue-green algae, chrysophytes and cryptophytes. 

Protozoans, including ciliates and flagellates, were also present, comprising up to 3 % of the 

assemblages in intake samples. Dominant taxa during these trials were Aulacoseira spp. 

(filamentous diatom), Melosira spp. (filamentous diatom), Cyclotella spp. (single-celled centric 

diatom), Asterionella formosa (colonial pennate diatom), Fragilaria spp. (filamentous diatom), 

Oscillatoria limnetica (filamentous blue-green), Pandorina morum (colonial green), and 

Rhodomonas spp. (single-celled cryptophyte). 

Microbial organism concentrations in the intake and control discharge samples during the 

SiCURETM trials are provided in table 6. 
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March 15, 2010 

Page 33 of 58 

Table 6. Microbial Organism Concentrations in Test Facility Intake Water and 

Control Discharge Across Trials. 

Microbial 

Organism

IMO G8 

Guideline Sample Trial A Trial B Trial C Trial D Trial E

E. Coli 

(MPN/100 mL) 

N/A Intake 31 213 51 9 8 

NA Control 

Discharge < 1 1.0 1.0 < 1 1.0 

Toxigenic V. 

cholerae 

(CFU/100 mL) 

N/A Intake < 1 < 1 < 1 < 1 < 1 

N/A Control 

Discharge < 1 < 1 < 1 < 1 < 1 

Enterococci 

(MPN/100 mL) 

N/A Intake 86 434 640 660 148 

N/A Control 

Discharge 35 2275 295 149 54 

Total 

Heterotrophic 

Bacteria 

(MPN/mL) 

N/A Intake 1217 8133 3667 2900 467 

NA Control 

Discharge 1650 1200 500 88 900 

 

3.2. Viable Organisms in Treated Discharge 

3.2.1 Regulated Plankton 

Using the methods noted in the methods section for determining the numbers of live organisms 

that fit within the regulated size categories of plankton, organism densities in discharge water 

from the five consecutive trials consistently met the IMO D-2 standard2

. The density of live 

organisms > 50 μm size class decreased dramatically immediately post-treatment (> 99.9 % 

reduction), and ranged from 38 to 193 individuals per m3. Bosminids, copepod copepodites, 

chironomids, dreissenid veligers, and a few Keratella and Lecanid rotifers were observed in the 

post-treatment samples. Soft-bodied rotifers were entirely absent from the subsamples analyzed. 

In post-retention discharge samples, the average density of live organisms in the > 50 μm size 

class ranged from 1 to 7 individuals per m3, .meeting G8 requirements (table 7). 

In the smaller size class, numbers of live organisms in the immediate post-treatment samples also 

were much lower than at intake, ranging from 1 to 13 per mL. Concentrations in discharge 

samples after the five day retention period were consistently less than 10 cells/ml, meeting G8 

requirements for treatment. 

 

2 As noted in the methods section, not all zooplankton in the natural freshwater assemblage meet the strict definition 

of 50 μm in minimum dimension. The total average density of live zooplankton in discharge samples ranged from 2 

to 20 per m3. The live organisms in these discharge samples were chironomid larvae, copepod nauplii, and Lecanid 

rotifers, with the latter being generally below the regulated size range as noted above. A list of all live zooplankton 

by taxonomic category and size class appear in Appendix 3.
GSI/LB/F/A/1 

March 15, 2010 

Page 34 of 58 

Table 7. Live Plankton Densities within Regulated Size Categories in Test Facility Post-Treatment 

Intake and Discharge Across Trials. 

Live Organism 

Size Category 

IMO G8 

Guideline Sample Trial A Trial B Trial C Trial D Trial E 

> 50 μm (#/m3) 

NA PostTreatment 193 38 151 176 53 

<10/m3 Discharge 5 7 4 4 1 

10 – 50 μm 

(#cells/mL) 

NA PostTreatment 6.8 1 5.8 4.8 13.5 

<10 /mL Discharge 0.0 0.0 0.0 6.8 2.9 

3.2.2. Regulated Microbes 

Immediate post-treatment, and post-treatment post-retention microbial samples were consistently 

below the IMO G8 standard for E. Coli and enterococcus, though intake levels and control 

discharge levels of E. coli were also below G8 discharge limits for all trials (table 8). 

Enterococcus levels in treated discharge were always 1 to 3 logs lower than control discharge 

and never above 10 MPN per 100 mL (table 8). Although heterotrophic plate counts immediately 

following treatment averaged an MPN fewer than ten per mL, post-treatment post-retention 

MPN average increased to 4.1E+05 per mL (table 8). This increase was not observed in the post 

retention control samples. 
GSI/LB/F/A/1 

March 15, 2010 

Page 35 of 58 

Table 8. Microbial Organism Concentrations in Test Facility Post-Treatment Intake 

and Discharge Across Trials. 

Microbial 

Organism

IMO G8 

Guideline Sample Trial A Trial B Trial C Trial D Trial E

E. Coli 

(MPN/100 mL) 

N/A 

PostTreatment 

Intake 

< 1 < 1 < 1 < 1 < 1 

< 250 CFU/ 

100 mL Discharge < 1 < 1 < 1 < 1 < 1 

Toxigenic V. 

cholerae 

(CFU/100 mL) 

N/A 

PostTreatment 

Intake 

< 1 < 1 < 1 < 1 < 1 

< 1 CFU 

V. cholerae/ 

100 mL 

Discharge < 1 < 1 < 1 < 1 < 1 

Enterococci 

(MPN/100 mL) 

N/A 

PostTreatment 

Intake 

< 1 < 1 < 1 < 1 < 1 

< 100 CFU/ 

100 mL Discharge < 1 3 < 1 2 1 

Total 

Heterotrophic 

Bacteria 

(MPN/mL) 

N/A 

PostTreatment 

Intake 

<2 <2 <2 4 5 

N/A Discharge 220,000 680,000 280,000 423,333 463,333 

 

3.3. Physical/Chemical Water Conditions 

Total residual oxidant (TRO) and total residual chlorine (TRC) levels in the pre- and posttreatment intake water, filter backwash, and control intake water are shown in table 9. As 

expected, levels of TRC and TRO in intake pre-treatment and control intake water were below 

regulatory concern. Also, as expected, in immediate post-treatment intake water samples the 

TRO and TRC were well above the allowable levels for discharge into receiving waters. Filter 

backwash, however, which was affected by low level electrolytic chlorination of the intake 

stream, was consistently below detection limits of TRC due to low initial levels, and rapid 

degradation of chlorine by high organic loads. 
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March 15, 2010 

Page 36 of 58 

Table 9. Average Total Residual Chlorine (TRC) and Total Residual Oxidant (TRO) Concentration 

(mg/L, ± std. dev.) from Trials A-E at Intake. 

Sample 

Location 

Trial A Trial B Trial C Trial D Trial E 

TRC TRO TRC TRO TRC TRO TRC TRO TRC TRO 

Control Intake 

(Tub 1) <0.008 0.031 <0.008 0.027 <0.008 0.032 <0.008 0.027 <0.008 0.030 

Pre-Treatment 

Intake (Tub 4) <0.008 0.035 <0.008 0.030 <0.008 <0.027 <0.008 0.033 <0.008 0.030 

Immediate Post 

Treatment 

(Siemens 

Sample Port) 

4.28 ± 

0.21 

4.20 

± 

0.36 

3.37 ± 

0.03 

3.36 ± 

0.07 

3.06 ± 

0.21 

3.08 ± 

0.14 

3.33 ± 

0.28 

3.31 ± 

0.11 

3.64 ± 

0.07 

3.49 ± 

0.07 

Immediate Post 

Treatment 

(GSI Sample 

Port) 

5.23 ± 

1.44 

5.08 

± 

0.75 

4.19 ± 

0.34 

4.04 ± 

0.45 

3.67 ± 

0.35 

4.02 ± 

0.72 

3.89 ± 

0.58 

4.04 ± 

0.50 

4.58 ± 

0.52 

4.53 ± 

0.50 

Immediate Post 

Treatment 

(Tub 6) 

1.99 2.29 1.66 1.75 1.31 1.45 1.85 1.87 1.96 1.84 

Filter Backwash 

(Sample Port) <0.008 - <0.008 - <0.008 - <0.008 - <0.008 - 

After five days retention in the 200 m3 tanks, treated discharge samples had significantly higher 

levels than control discharge samples, but TRC levels in treated discharge was always 0.070 

mg/L or less (table 10). 

Table 10. Average Total Residual Chlorine (TRC) and Total Residual Oxidant (TRO) Concentration 

(mg/L, std. dev.) from Trials A-E on Discharge. 

Sample 

Location 

Trial A Trial B Trial C Trial D Trial E 

TRC TRO TRC TRO TRC TRO TRC TRO TRC TRO 

Treatment 

Retention 

Tank (Day 5, 

Prior to 

Discharge) 

0.058 0.084 0.064 0.065 0.055 0.062 0.070 0.070 0.069 0.064 

Control 

Discharge 

(Tub 1) 

<0.008 <0.027 <0.008 <0.027 <0.008 0.028 <0.008 <0.027 <0.008 <0.027 

Treatment 

Discharge 

(Tubs 4, 5, 

and 6) 

0.063 ± 

0.003 

0.086 ± 

0.002 

0.069 ± 

0.002 

0.071 ± 

0.001 

0.058 ± 

0.002 

0.065 ± 

0.003 

0.071 ± 

0.001 

0.078 ± 

0.004 

0.064 

± 

0.006 

0.083 ± 

0.008 
GSI/LB/F/A/1 

March 15, 2010 

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3.4. Residual Toxicity 

GSI conducted Cold and Standard WET assays for each trial in the five valid trials series. The 

results showed fairly consistent effects with respect to residual TRC and TRO concentrations 

across trials. There was no evidence of acute toxic effects even in the 100 percent effluent 

samples. Outcomes of chronic biological toxicity assays varied somewhat across trials and 

species within each assay in the undiluted effluent samples. However, results consistently 

showed no toxicity in the more diluted samples. 

3.4.1. TRC and TRO Concentrations in WET Tests 

Across assays, in 100 percent effluent, TRC concentrations were higher in the Cold WET 

solutions, than Standard WET solutions. For the P. promelas and C. dubia assays, the mean TRC 

concentration was 0.032 - 0.040 mg/L in the Standard WET assays, and 0.088-0.118 mg/L, in the 

Cold WET assays (table 11). In the S. capricornutum tests, the initial TRC concentrations 

ranged from 0.025 - 0.044 mg/L in the Standard WET assays, and from 0.097 - 0.134 mg/L in 

the Cold WET tests. In all tests TRC concentrations were below the limit of quantification 

(LOQ) in dilutions of whole effluent (tables 11 and 12). 

The range of TRO concentrations also differed between Standard WET and Cold WET solutions. 

In 100 % effluent across trials mean TRO concentrations for the P. promelas and C. dubia

ranged from 0.060 - 0.070 mg/L in Standard WET solutions, and 0.110 – 0.138 mg/L in Cold 

WET solutions (table 13). The initial TRO concentrations across trials for the S. capricornutum

bioassays ranged from 0.045 - 0.060 mg/L (table 13). In Cold WET solutions they were 0.108-

0.127 mg/L. 

Table 11. Mean (± Std. Err.) Total Residual Chlorine (TRC) Concentration (mg/L) of Renewal Water 

for Each Trial of P. promelas and C. dubia WET Bioassays. 

 Exposure 

Standard Whole Effluent Toxicity Bioassay Cold WET

Trial PPW CDW 0% 6.25% 12.5% 25% 50% 100% 100% 

A 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

0.040 

(0.001) 

0.102 

(0.003) 

B 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

0.032 

(0.002) 

0.094 

(0.005) 

C 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

0.034 

(0.002) 

0.088 

(0.005) 

D 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

0.039 

(0.002) 

0.108 

(0.005) 

E 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

<0.008 

(NA) 

0.039 

(0.001) 

0.118 

(0.006) 

 NA-Not appropriate. If one or more value was less than 0.008 mg/L this made calculating a meaningful a 

standard error inappropriate. PPW – Fathead minnow reference control. CDW – C. dubia reference 

control. 
GSI/LB/F/A/1 

March 15, 2010 

Page 38 of 58 

Table 12. Total Residual Chlorine (TRC) Concentration (mg/L) of the Initial Exposure Solution 

for Each Trial of S. capricornutum WET Bioassays. 

 Exposure 

Standard Whole Effluent Toxicity Bioassay 

Cold 

WET 

Trial SCW 0% 6.25% 12.50% 25% 50% 100% 100% 

A <0.008 <0.008 <0.008 <0.008 <0.008 <0.008 0.044 0.097 

B <0.008 <0.008 <0.008 <0.008 <0.008 <0.008 0.025 0.105 

C <0.008 <0.008 <0.008 <0.008 <0.008 <0.008 0.037 0.101 

D <0.008 <0.008 <0.008 <0.008 <0.008 <0.008 0.039 0.134 

E <0.008 <0.008 <0.008 <0.008 <0.008 <0.008 0.036 0.117 

SCW – S. capricornutum (green algae) reference control. 

Table 13. Mean (± Std. Err.) Total Residual Oxidant (TRO) Concentration (mg/L) of Renewal Water 

for Each Trial of P. promelas and C. dubia WET Bioassays. 

 Exposure 

Standard Whole Effluent Toxicity Bioassay 

Cold

WET 

Trial PPW CDW 0% 6.25% 12.5% 25% 50% 100% 100% 

A 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

0.038 

(0.003) 

0.060 

(0.006) 

0.118 

(0.008) 

B 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

0.030 

(NA) 

0.039 

(0.002) 

0.062 

(0.005) 

0.110 

(0.013) 

C 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

0.027 

(NA) 

0.040 

(0.002) 

0.062 

(0.002) 

0.109 

(0.011) 

D 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

0.029 

(NA) 

0.038 

(0.002) 

0.067 

(0.003) 

0.121 

(0.012) 

E 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

<0.027 

(NA) 

0.045 

(0.002) 

0.070 

(0.004) 

0.138 

(0.011) 

NA-Not appropriate. If one or more value was less than 0.027 mg/L this made calculating a 

meaningful a standard error inappropriate. PPW – Fathead minnow reference control. CDW – 

C. dubia reference control. 

 
GSI/LB/F/A/1 

March 15, 2010 

Page 39 of 58 

 Table 14. Total Residual Oxidant (TRO) Concentration (mg/L) of the Initial 

Exposure Solution for Each Trial of S. capricornutum Bioassays. 

 Exposure 

Standard Whole Effluent Toxicity Bioassay 

Cold 

WET 

Trial SCW 0% 6.25% 12.5% 25% 50% 100% 100% 

A <0.027 <0.027 <0.027 <0.027 0.034 0.037 0.060 0.114 

B 0.036 <0.027 <0.027 <0.027 0.029 0.032 0.045 0.127 

C 0.036 <0.027 <0.027 <0.027 0.030 0.033 0.060 0.124 

D 0.033 <0.027 <0.027 <0.027 <0.027 0.042 0.057 0.123 

E <0.027 <0.027 <0.027 <0.027 0.030 0.040 0.054 0.108 

SCW – S. capricornutum (green algae) reference control. 

3.4.2. Whole Effluent Toxic Effects 

3.4.2.1. Standard WET Results 

In the Standard WET tests, no statistically significant acute toxic effect (i.e., reduction in 

survival) was detected across trials and test species (tables 15, 17, and 18). With respect to 

chronic toxicity in these tests, no effects were detected at any effluent dilution relative to 

exposed fish P. promelas (table 15), or any other test species at 50 % effluent dilution or below. 

Phytoplankton (Selenastrum) assays for two trials (Trials A and E) met data quality criteria.3

 In 

Trial A, an estimated 77 % effluent dilution appeared sufficient to reduce algal population 

growth by 25 %, while in Trial E no reduction in cell growth was observed even in 100 % 

effluent (table 16). Chronic (i.e., reproductive) effects in the zooplankton species C. dubia were 

observed in 100 % effluent in Standard WET tests in one out of two trials (tables 17 and 19; 

Trial A; p< 0.05). An estimated 43 % effluent dilution in Trial A, would reduce the mean 

number of young produced per female by 25 % (table 17). No chronic toxicity was observed in 

the other valid trial (Trial B). 

3.4.2.2. Cold WET Results 

The Cold WET assays involved only 100 percent effluent solutions, and measured TRC and 

TRO values were significantly higher than in the Standard WET assays. Fathead minnow was 

unaffected across five trials. Acute toxicity (increased mortality) of C. dubia was measured in 

one out of two valid trials (table 18; Trial A; p<0.05). There was chronic toxicity (reduced 

reproduction) detected in C. dubia in the same trial (table 19; Trial A; p<0.05). No such effects 

on C. dubia were detected in the other valid trial (tables 18 and 19; Trial B). In addition, 100 % 

effluent samples had no chronic toxicity effect on the algal species Selenastrum (data not 

shown). 

 

3 Starting densities exceeded GSI standard operating procedure (SOP is based on US EPA methods). 
GSI/LB/F/A/1 

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Table 15. Effective Concentration in 25 % of Test Organisms (EC25), No-Observed-EffectConcentration (NOEC) and Lowest-Observed-Effect-Concentration (LOEC) for Pimephales 

promelas Measured in terms of Reduced Survival or Growth of the Test Individuals when Exposed 

to Effluent during Standard WET Tests. 

Acute Toxicity (%Effluent) Chronic Toxicity (%Effluent) 

Trial LC50 (95% CI) NOEC LOEC EC25 (95% CI) NOEC LOEC 

A >100 100 >100 >100 100 >100 

B >100 100 >100 >100 100 >100 

C >100 100 >100 >100 100 >100 

D >100 100 >100 >100 100 >100 

E >100 100 >100 >100 100 >100 

Table 16. Effective Concentration in 25 % of Test Organisms (EC25), No-Observed-EffectConcentration (NOEC) and Lowest-Observed-Effect-Concentration (LOEC) for Selenastrum 

capricornutum Measured in terms of Reduced Cell Growth when Exposed to Effluent during 

Standard WET Tests. 

a Inadequate survival of or too much variation in control samples.

Chronic Toxicity (%Effluent) 

Trial EC25 (95% CI) NOEC LOEC 

A 77 (N/A) 50 100 

B

a

- - - 

C

a

- - - 

D

a

- - - 

E >100 100 >100 
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Table 17. Effective Concentration in 25 % of Test Organisms (EC25), No-Observed-EffectConcentration (NOEC) and Lowest-Observed-Effect-Concentration (LOEC) for Ceriodaphnia dubia

Measured in terms of Reduced Survival or the Mean Number of Young Produced per Female when 

Exposed to Effluent during Standard WET Tests. 

Acute Toxicity (%Effluent) Chronic Toxicity (%Effluent) 

Trial LC50 (95% CI) NOEC LOEC EC25 (95% CI) NOEC LOEC 

A >100 100 >100 43 (32-70) 50 100 

B >100 100 >100 >100 100 >100 

C

a

- - - - - - 

D

c

- - - - - - 

E

c

- - - - - - 

a

 Inadequate survival in the control treatment group. 

c

 Inadequate reproduction rate per female in controls. 

Table 18. Average (n=10) Percent Mortality (Standard Error) of Ceriodaphnia dubia. 

a

 Inadequate survival in the control treatment group. 

c

 Inadequate reproduction rate per female in controls. 

* Statistically significant (p<0.05) 

Trial Control 6.25 12.5 25 50 100 Cold WET 

100% 

A 0 (0) 0 (0) 0 (0) 0 (0) 10 (6) 20 (8) 50 (16)* 

B 10 (6) 10 (6) 10 (6) 0 (0) 0 (0) 0 (0) 0 (0) 

C

a

- - - - - - - 

D

c

- - - - - - - 

E

c

- - - - - - - 
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 Table 19. Average (n=10) Number of Young per Female (standard error) of Ceriodaphnia dubia. 

a

 Inadequate survival in the control treatment group. 

c

 Inadequate reproduction rate per female in controls. 

* Statistically significant when compared to control (p<0.05) 

3.4.3. Disinfection By-Products (DBPs) 

Levels of all analyzed compounds were below the method reporting limits (see table 20 for 

MRLs) in all control water samples collected during Test Trials A-E. During Test Trials A-E, 

the only measureable concentrations of DBPs found in the backwash samples were for several 

samples containing sodium chlorate ranging in concentration from 12 to 13 μg/L (table 20). 

Measureable levels of many of the THMs and HAAs were found in the treated water samples 

collected immediately post treatment and from samples collected after the five day retention 

time. Total THM concentrations were higher in the samples collected after the five-day retention 

time than found in the samples collected immediately post treatment in all cases. Total THM 

concentrations ranged from 101 to 192 μg/L in post-treatment sample water and 229 to 265 μg/L 

after five-day retention (table 20). Total HAAs ranged from 98 to 180 μg/L in post-treatment 

and treated discharge sample water and displayed no definite pattern with time (table 20). 

Dichloroacetonitrile was the only haloacetonitrile compound out of four that were analyzed that 

had measureable concentrations in any of the water samples collected. Dichloroacetonitrile was 

measured only in water samples collected immediately post treatment, and concentrations ranged 

from 2.9 to 4.1 μg/L (table 20), but this compound was below the method reporting limit in 

treated water after a five-day retention. Sodium chlorate concentrations in treated water 

collected immediately post treatment and on discharge, ranged from 370 to 450 μg/L (table 20). 

Sodium bromate was not detected in any of the samples collected during intake or discharge. 

Trial Control 6.25 12.5 25 50 100 Cold WET 

100% 

A 21 (1.5) 22 (0.9) 21 (1.2) 23 (1.0) 15 (1.5) 12 (0.9)* 5 (1.4)* 

B 21 (1.0) 18 (1.7) 17 (1.4) 23 (0.8) 24 (0.6) 17 (0.8) 20.9 (1.8) 

C

a

- - - - - - - 

D

c

- - - - - - - 

E

c

- - - - - - - 
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Table 20. Results from Analysis of Disinfection By-Products Measured in Samples Collected 

during Siemens SiCURETM Performance Evaluations. 

Note: Te method reporting limits (MRLs) for each class of chemicals is the summation of all the 

individual reporting limits for each chemical, and are as follows: trihalomethanes MRL = 2.0 μg/L, 

haloacetic acids MRL = 11.0 μg/L, dichloroacetonitrile MRL = 0.5 μg/L, and sodium chlorate MRL = 

10.0 μg/L. 

Parameter Sample Trial A Trial B Trial C Trial D Trial E 

Trihalomethanes 

(μg/L) 

Intake - Filter 

Backwash Line <2.0 <2.0 <2.0 <2.0 <2.0

Post-Treatment 

(Tub 6) 100.5 123.8 121.4 191.7 161.6 

Control Discharge 

(Tub 1) <2.0 <2.0 <2.0 <2.0 <2.0

Treatment 

Discharge (Tub 6) 229.4 260.8 264.7 233.6 264.2 

Haloacetic Acids 

(μg/L) 

Intake - Filter 

Backwash Line <11.0 b <4.0 <11.0 <11.0 <11.0

Post-Treatment 

(Tub 6) 127.2 

a Not 

Measured 123.8 158.1 156.4 

Control Discharge 

(Tub 1) <11.0 

a Not 

Measured <11.0 <11.0 <11.0

Treatment 

Discharge (Tub 6) 180.4 101.5 97.8 117.2 104.1 

Sodium chlorate 

(μg/L) 

Intake - Filter 

Backwash Line <10.0 <10.0 13 12 13 

Post-Treatment 

(Tub 6) 420 380 370 450 400 

Control Discharge 

(Tub 1) <10.0 <10.0 <10.0 <10.0 <10.0

Treatment 

Discharge (Tub 6) 420 380 380 440 410 

Dichloroacetonitrile

(μg/L) 

Intake - Filter 

Backwash Line <0.5 <0.5 <0.5 <0.5 <0.5

Post-Treatment 

(Tub 6) 4.1 3.7 3.2 3.2 2.9 

Control Discharge 

(Tub 1) <0.5 <0.5 <0.5 <0.5 <0.5

Treatment 

Discharge (Tub 6) <0.5 <0.5 <0.5 <0.5 <0.5

a Sample bottles broke in transit to the laboratory. 

b

 Zeros (not MRL values) were reported for all haloacetic acids analyzed, with the exception of 

tribromoacetic acid for which the MRL of <4.0 μg/L was reported. 
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4.0. DISCUSSION OF RESULTS 

The GSI tests on the SiCURETM Ballast Water Management System were comprehensive and 

informative of the treatment system’s potential performance relative to IMO G8 guidelines in 

fresh water systems like the Great Lakes. The SiCURETM Ballast Water Management System 

operated without interruption during five consecutive valid trials, and biological performance 

outcomes were remarkably consistent across trials. Treated discharge water from these valid 

trials contained densities of organisms within the regulated size classes that were below the IMO 

D-2 standards, as determined using methods documented in this report. Though data on V. 

cholera concentration are still pending, other microbial analyses showed system performance in 

keeping with IMO requirements for bacteria. Chemistry data generated across trials indicated 

the post-retention discharge to have well less than 0.1 mg/L total residual chlorine under ambient 

conditions. Ambient water collected immediately after treatment and held in a cold environment 

had TRC and TRO levels which slightly exceeded this level. However, in a real world 

application, the intake water would also be cold, and developers claim that the test system is 

designed to respond to this circumstance (reflected in oxidation-reduction potential, or ORP) 

with a reduction in chlorine generated and injected into the intake stream.

Disinfection by-products were detectable in post-treatment and post-retention water. Little 

aquatic chronic toxicity data are available for the DBPs tested. A comparison to United States 

Federal Drinking Water Maximum Contaminant Levels (MCLs) show the total THM values in 

the treated water above the limits for total THMs (80 μg/L) and HAA (60 μg/L) in all of the 

treated water samples. However, this comparison is very conservative as it pertains to drinking 

water not subject to dilution effects. 

There were two to five valid replicates of Standard and Cold WET assays for all organisms 

across trials. There were no statistically significant (p<0.05) acute toxic effects of treated 

discharge on any test species across assays and trials. There were no statistically significant 

(p<0.05) detectable chronic toxicity effects across organisms and trials in 50 percent or lower 

effluent dilutions. Chronic toxicity effects (reduced reproduction) in 100 percent effluent were 

detected in Trail A for test species of zooplankton and possibly phytoplankton in both the 

Standard and Cold WET assay, but not in other trials or dilution. 
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5.0. CONCLUSIONS 

The Siemens SiCURETM Ballast Water Management System functioned properly during five 

consecutive trials, and was highly effective at reducing live organism densities in the fresh water 

ambient conditions of Duluth-Superior Harbor, as amended in these tests to achieve IMOconsistent challenge conditions. Live organisms in the regulated size classes were discharged in 

densities below the IMO D-2 standard. Further data and testing are needed to fully characterize 

residual toxicity after a five day retention period, but we detected no consistent or pronounced 

effect in these trials. 

REFERENCES 

GSI (2009). Quality Assurance Project Plan for Great Ships Initiative (GSI) Bench-Scale and 

Land-Based Biological Tests (2009). 

 

Hasan JA, Huq A, Tamplin ML, Siebeling RJ and Colwell RR (1994). A Novel Kit for Rapid 

Detection of Vibrio cholerae O1. Clinical Microbiology; 32(1): 249-252 

International Maritime Organization (IMO), 2004. International Convention for the Control and 

Management of Ships Ballast Water and Sediments. As adopted by consensus at a Diplomatic 

Conference at IMO, London, England, February 13 2004. 

U.S. EPA, Office of Water (2002). Short-term Methods for Estimating the Chronic Toxicity of 

Effluents and Receiving Waters to Freshwater Organisms, 4th Edition. Accessed from: 

http://www.epa.gov/waterscience/methods/wet/disk3/index.html. 

APPENDICES 

1. Design of the GSI Land-Based RDTE Facility 

2. GSI SOPs Relevant to SiCURETM Tests 

3. Average Density of Live Zooplankton in Treatment Discharge 

4. Data Quality Objectives, Criteria and Results 

5. Operational Data Summary 
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APPENDIX 1. 

GSI LAND-BASED RDTE FACILITY: TECHNICAL INFORMATION 

The GSI’s Land-Based Research, Development and Technology Evaluation (RDTE) Facility in 

Superior, Wisconsin is used to conduct full-scale biological evaluations of prospective ballast 

treatments suitable to Seaway-sized vessels. The facility draws raw intake water and entrained 

organisms from Duluth-Superior Harbor at up to 680 m

3

/hr. After initial transport through 16 

inch HDPE line to the facility, a carefully designed “Y-split” in the intake piping simultaneously 

channels one half of the flow (up to 340 m3

/hr) to a treatment track and one half (up to 340 

m

3

/hr) to a matched control track (figure 1). Water in the treatment track passes through the 

experimental ballast treatment system and into one of the 200 m3 cylindrical treatment retention 

tanks (test tank #1 or #2; figure 1). Water in the control track by-passes the treatment system and 

is channeled directly into a matched control retention tank (control tank #1 or #2; figure 1). After 

storage (duration dependent on test requirements), the water is discharged sequentially from the 

treatment and control retention tanks at up to 340 m

3

/hr. Depending on the test scenario, the 

water is either discharged to the harbor or transferred to an alternate retention tank for 

subsequent transport to a wastewater facility, or through the treatment system again for discharge 

or retention. 

Treatment and control intake and discharge water is sampled at pressure/flow controlled in-line 

sample points (SPs). Intake samples are collected concurrently on the control and treatment 

tracks respectively (using SP#2 and SP#3, figure 2). Post-treatment samples are collected from 

SP#15 (figure 2). Discharge samples are collected from one of two discharge sample points 

(SP#9, or SP#10; figure 2), with sequential sampling of control and treatment water. At each of 

these SPs there are three replicate sample ports with a center-located 3.8 cm internal diameter 

(ID) elbow-shaped pitot tube (figure 3) connected to a 3.8 cm ID PVC transfer pipe that carries 

the sample water to one of six 3.8. m3

 collection tubs located at a centralized sampling station 

(figure 2). Injection Ports A and B (figure 1) can be used to meter fine grade Arizona Test Dust, 

and concentrated algae harvested from the Duluth-Superior Harbor into the intake stream to 

assure adequate concentrations of Total Suspended Solids (TSS) and live phytoplankton in intake 

and control discharge. Other SPs shown in figure 2, with one port per SP, are used for calibration 

testing the facility itself and not typically used for sample collection during a treatment system 

evaluation. 

A mobile field laboratory and on-site stationary structure provides bench-scale facilities to 

support time-sensitive assays associated with tests conducted at the GSI Land-Based RDTE 

Facility. The mobile laboratory is located at the facility during testing but may be moved to other 

sites in the Great Lakes-St. Lawrence Seaway System to support GSI shipboard tests when 

required. Both laboratories are climate-controlled with enough desk and counter space to allow 

for simultaneous microscopic and analytical analysis of samples. In addition, laboratories of the 

University of Wisconsin-Superior’s Lake Superior Research Institute (LSRI) and the University 

of Minnesota-Duluth’s Natural Resources Research Institute provide non-time sensitive analysis 

of samples from the land-based tests. Since both facilities are only a few miles from the facility, 

samples can be easily transported for rapid analysis. 
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March 15, 2010 

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Figure 1. Simplified Schematic of the GSI Land-Based RDTE Facility. 
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Figure 2. Schematic of the GSI Land-Based RDTE Facility Showing the Location of the Intake and 

Discharge Sample Points (i.e., SP #s 2, 3, 9, and 10), and Corresponding Sample Collection Tubs. 

Figure 3. Schematic of a Sample Point (SP) Showing the Design of the 

Three Sample Port Pitots. 
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APPENDIX 2. 

GSI SOPs Relevant to Siemens SiCURETM Performance Evaluation 

SOP/Analysis 

Category SOP ID Code SOP Title Date of Issue 

Health and Safety GSI/SOP/LB/G/S/1 Procedure for Ensuring Worker Health and Safety at 

the GSI Land-Based RDTE Facility June 26, 2009 

Facility Operation GSI/SOP/LB/G/O/1 Procedure for Operating the GSI Land-Based RDTE 

Facility July 20, 2009 

Facility Operation GSI/SOP/LB/G/O/3 Procedure for Cleaning the Retention Tanks and Other 

Equipment at the GSI Land-Based RDTE Facility July 20, 2009 

Facility Operation GSI/SOP/LB/G/O/5 Procedure for Injecting Organisms and Solids into the 

GSI Land-Based RDTE Facility September 1, 2009 

Facility Operation GSI/SOP/LB/G/O/7 Procedure for Solids Resuspension After Retention at 

the GSI Land-Based RDTE Facility DRAFT 

Sample Handling 

and Custody GSI/SOP/G/RA/SC/3 Procedure for Labeling Samples Collected at the GSILand-Based RDTE Facility July 15, 2009 

Water Chemistry GSI/SOP/BS/RA/C/2 Procedure for Determining Total Residual Oxidants 

(TRO) in Water July 7, 2009 

Water Chemistry GSI/SOP/BS/RA/C/6 Procedure for Analyzing Total Residual Chlorine 

Concentrations in Water July 15, 2009 

Water Quality GSI/SOP/BS/RA/C/3 Procedures for Measuring Organic Carbon in Aqueous 

Samples May 28, 2009 

Water Quality GSI/SOP/BS/RA/C/8 Procedure for Analyzing Total Suspended Solids (TSS) June 30, 2009 

Sample Collection GSI/SOP/LB/RA/SC/8 Procedure for Collecting Physical/Chemical Data and

Samples at the GSI Land-Based RDTE Facility. DRAFT 

Sample Collection GSI/SOP/LB/RA/SC/1 Procedure for Collecting Biological Sample Water via 

In-Line Sample Ports July 20, 2009 

Sample Collection GSI/SOP/LB/RA/SC/6 Procedure for Zooplankton Sample Collection July 20, 2009 

Zooplankton 

Sample Analysis GSI/SOP/LB/RA/SA/2 Procedure for Zooplankton Sample Analysis July 20, 2009 
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Sample Collection GSI/SOP/LB/RA/SC/3 Procedure for Algae/Small Protozoa Sample Collection June 26, 2009 

Phytoplankton 

Sample Analysis GSI/SOP/LB/RA/SA/1 Procedure for Algae/Small Protozoan Sample Analysis June 26, 2009 

Sample Collection GSI/SOP/LB/RA/SC/4 Procedure for Microbial Sample Collection July 16, 2009 

Microbial Sample 

Analysis GSI/SOP/LB/RA/SA/3 Procedure for the Colony Blot Preparation for 

Enumeration of Culturable Vibrio chloreae DRAFT 

Microbial Sample 

Analysis GSI/SOP/BS/RA/MA/1 Procedure For Quantifying Heterotrophic Plate Counts(HPCs) Using IDEXX’s SimPlate® for HPC Method July 17, 2009 

Microbial Sample 

Analysis GSI/SOP/BS/RA/MA/3 Procedure for the Detection and Enumeration of 

Enterococcus using EnterolertTM July 17, 2009 

Microbial Sample 

Analysis GSI/SOP/BS/RA/MA/4 Procedure for the Detection and Enumeration of Total 

Coliforms and E. coli Using IDEXX's Colilert® July 17, 2009 

Whole Effluent 

Toxicity Testing GSI/SOP/BS/RA/WET/1 Procedure for Assessing Chronic Residual Toxicity of a 

Ballast Treatment System to Ceriodaphia dubia July 16, 2009 

Whole Effluent 

Toxicity Testing GSI/SOP/BS/RA/WET/2 

Procedure for Assessing Chronic Residual Toxicity of a 

Ballast Treatment System to the Fathead Minnow 

(Pimephales promelas)

July 16, 2009 

Whole Effluent 

Toxicity Testing GSI/SOP/BS/RA/WET/3 

Procedure for Assessing Chronic Residual Toxicity of a 

Ballast Treatment System to the Green Alga 

(Selenastrum capricornutum) 

DRAFT 

Data Management GSI/SOP/G/RA/DM/1 Procedure for Data Entry, Data Quality Control and 

Database Management 

Available on request 

(Internal document) 
GSI/LB/F/A/1 

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APPENDIX 3. 

Average Density (per m3) of Live Zooplankton in Treatment Discharge during

the Trials of the SiCURETM System. 

Test Trials: Trial A Trial B Trial C Trial D Trial E 

Total Vol. Treatment Discharge Analyzed, m3: 3.84 3.96 6.11 5.97 6.05 

Greater than 50 μm (min. dimension)

Taxa Group Species 

Avg. 

Density 

(per m3) 

Avg. 

Density 

(per m3)

Avg. 

Density 

(per m3)

Avg. 

Density 

(per m3)

Avg. 

Density 

(per m3)

Arachnida Mite 0.17 

Cladocerans Bosmina 0.17 

Daphnia 0.17 

Copepods Copepod 0.79 0.17 

Diptera Chironomid 2.08 4.03 2.79 2.36 0.66 

Nematoda Nematode 0.33 

Ostracoda Ostracod 0.17 

Dreissena Dreissenid (Zebra Mussel) 0.50 

Nauplii Copepod Nauplii 1.56 2.52 1.32 0.34 0.66 

Tardigrade Tardigrade 0.49 

Rotifers Bdelloid 0.52 0.33 

Keratella 0.33 

Greater than 50 μm (min. dimension) Total: 4.95 7.54 4.44 4.04 1.49 

Less than 50 μm (min. dimension)

Taxa Group Species 

Avg. 

Density 

(per m3)

Avg. 

Density 

(per m3)

Avg. 

Density 

(per m3)

Avg. 

Density 

(per m3)

Avg. 

Density 

(per m3)

Rotifers Lecanidae 5.21 12.57 7.52 3.02 0.99 

Less than 50 μm (min. dimension) Total: 5.21 12.57 7.52 3.02 0.99 
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APPENDIX 4. DATA QUALITY OBJECTIVES, CRITERIA 

AND RESULTS 

Table 1. Water Quality Analyses. 

Data Quality 

Indicator 

Evaluation Process/ 

Performance Measurement 

Data Quality 

Objective Performance Measurement Result 

Precision 

Samples (10%) are collected 

and analyzed in duplicate – with 

performance measured by 

average relative percent 

difference (RPD) of all duplicate 

analyses performed during test 

trials. 

<20% average 

(± SD) RPD. 

Percentage of Water 

Quality Samples 

Collected and 

Analyzed in Duplicate: 

17%

Total Suspended Solids: 4.5% 

(±6.7%) RPD, n=6 

Dissolved Organic Carbon: 

2.4% (±1.8) RPD, n=6 

^Particulate Organic Carbon:

111.4% (±100.6%) RPD, n=6 

Accuracy 

Performance is measured by 

average percent difference 

(%D) between all measured and 

nominal reference standard 

values from test trials. 

<20% average 

(± SD) D. 

Percentage of Analysis 

Days Containing a 

Reference Standard: 

100%

Total Suspended Solids: 4.3% 

(±2.7%) D, n=5 

Dissolved Organic Carbon: 

1.0% (±1.0) D, n=5 

Representativeness 

Pre-treatment/control and 

post-treatment/treatment 

samples are handled and 

analyzed in the same manner. 

Not Applicable 

– Qualitative. 

All pre-treatment and post-treatment water quality 

intake samples, as well as all control and treatment 

discharge samples, were collected, handled, and 

analyzed in the same manner (using the appropriate GSI 

SOPs). 

Comparability 

Routine procedures are 

conducted according to 

appropriate SOPs to ensure 

consistency between tests. 

Not Applicable 

– Qualitative. 

The following GSI SOPs were used for all water quality 

analyses conducted during the test trials: 

- GSI/SOP/BS/RA/C/3 – Procedures for Measuring 

Organic Carbon in Aqueous Samples 

- GSI/SOP/BS/RA/C/8 – Procedure for Analyzing Total 

Suspended Solids (TSS) 

Completeness 

Number of valid data obtained 

from the performance 

measurement system vs. 

number performance 

measurement samples 

analyzed. 

Performance is measured by 

percent completeness (%C). 

>90% C. 

Total Suspended Solids: 10/10 performance 

measurements met DQO = 100% C

Dissolved Organic Carbon: 10/10 performance 

measurements met DQO = 100% C

^Particulate Organic Carbon: 2/5 performance 

measurements met DQO = 40% C

^Water quality parameter having performance measurement result that did not meet data quality objective 

specified in GSI QAPP (2009). 
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Table 2. Chemistry Analyses. 

Data Quality 

Indicator 

Evaluation Process/ 

Performance Measurement 

Data Quality 

Objective Performance Measurement Result 

Precision 

Samples (10%) are collected 

and analyzed in duplicate – 

with performance measured 

by average relative percent 

difference (RPD) of all 

duplicate analyses performed 

during test trials. 

<20% average 

(± SD) RPD. 

Percentage of 

Chemistry Samples 

Collected and Analyzed 

in Duplicate: 20%

Total Residual Chlorine: 4.8% 

(±5.3%) RPD, n=11 

Total Residual Oxidants: 8.2% 

(±7.0%) RPD, n=11 

Bias 

Performance is measured by 

average percent spikerecovery (%SPR) of all 

analyses performed during 

test trials.

75%-110% 

average (± SD) 

SPR. 

Percentage of 

Chemistry Samples 

Collected, Spiked, and 

Analyzed: 18% (TRC)

and 15% (TRO)

Total Residual Chlorine: 98.6% 

(±15.5%) RPD, n=10 

Total Residual Oxidants: 95.7% 

(±3.8%) RPD, n=8 

Accuracy 

Performance is measured by 

average percent difference 

(%D) between all measured 

and nominal reference 

standard values from test 

trials. 

<20% average 

(± SD) D. 

Percentage of Analysis 

Days Containing a 

Reference Standard: 

100%

Total Residual Chlorine: 4.8% 

(±3.0%) D, n=10 

Total Residual Oxidants: 7.2% 

(±1.1%) D, n=10 

Representativeness 

Pre-treatment/control and 

post-treatment/treatment 

samples are handled and 

analyzed in the same manner. 

Not Applicable – 

Qualitative. 

All pre-treatment and post-treatment water quality intake 

samples, as well as all control and treatment discharge 

samples, were collected, handled, and analyzed in the same 

manner (using the appropriate GSI SOPs). 

Comparability 

Routine procedures are 

conducted according to 

appropriate SOPs to ensure 

consistency between tests. 

Not Applicable – 

Qualitative. 

The following GSI SOPs were used for all water quality 

analyses conducted during the test trials: 

-GSI/SOP/BS/RA/C/2 – Procedure for Determining 

Total Residual Oxidants (TRO) in Water 

-GSI/SOP/BS/RA/C/6 – Procedure for Analyzing 

Total Residual Chlorine (TRC) Concentrations in 

Water 

Completeness 

Number of valid data obtained 

from the performance 

measurement system vs. 

number performance 

measurements collected. 

Performance is measured by 

percent completeness (%C). 

>90% C. 

Total Residual Chlorine: 29/31 performance measurements 

met DQO = 94% C

Total Residual Oxidants: 28/29 performance measurements 

met DQO = 97% C

Sensitivity 

The limit of detection (LOD) 

and quantification (LOQ) for 

the analytical method used is 

reported. 

Dependent upon 

the analyte and 

instrumentation. 

Total Residual Chlorine: LOD = 3 μg/L; LOQ = 8 μg/L

Total Residual Oxidants: LOD = 8 μg/L; LOQ= 27 μg/L
GSI/LB/F/A/1 

March 15, 2010 

Page 54 of 58 

Table 3. Phytoplankton (Organisms <50 μm) Analyses. 

Data Quality 

Indicator 

Evaluation Process/ 

Performance Measurement 

Data Quality 

Objective Performance Measurement Result 

Bias 

Samples (10%) are analyzed by 

two separate taxonomists – 

with performance measured by 

average percent similarity (PS)

of taxonomic identification (live 

entities) and average relative 

percent difference (RPD) of the 

number of live entities counted 

for all second (QA) analyses 

performed during test trials. 

>60% average 

(± SD) PS and 

<20% average 

(± SD) RPD. 

Percentage of 

Phytoplankton 

Samples Analyzed by a 

Second Taxonomist: 

35%

97.5% (± 2.1%) PS, n=5 

3.30% (± 3.27) RPD, n=5 

Representativeness 

Pre-treatment/control and 

post-treatment/treatment 

samples are handled and 

analyzed in the same manner. 

Not Applicable 

– Qualitative. 

All pre-treatment and post-treatment phytoplankton 

samples collected on intake, as well as all control and 

treatment discharge samples, were collected, handled, 

and analyzed in the same manner (using the appropriate 

GSI SOPs). 

Comparability 

Routine procedures are 

conducted according to 

appropriate SOPs to ensure 

consistency between tests. 

Not Applicable 

– Qualitative. 

The following GSI SOP was used for all phytoplankton 

sample analyses conducted during the test trials: 

- GSI/SOP/LB/RA/SA/1 – Procedure for Algae/Small 

Protozoan Sample Analysis 

Completeness 

Number of valid data obtained 

from the performance 

measurement system vs. 

number performance 

measurement samples 

analyzed. 

Performance is measured by 

percent completeness (%C). 

>90% C. 9/10 performance measurements met DQO = 90% C
GSI/LB/F/A/1 

March 15, 2010 

Page 55 of 58 

Table 4. Zooplankton (Organisms >50 μm) Analyses. 

Data Quality 

Indicator 

Evaluation Process/ 

Performance Measurement 

Data Quality 

Objective Performance Measurement Result 

Bias 

Samples (10%) are analyzed by 

two separate taxonomists – 

with performance measured by 

average percent similarity (PS)

of taxonomic identification (live 

organisms only) and average 

relative percent difference 

(RPD) of the number of live 

organisms counted for all 

second (QA) analyses 

performed during test trials. 

>90% average 

(± SD) PS and 

<20% average 

(± SD) RPD. 

Percentage of 

Zooplankton Samples 

Analyzed by a Second 

Taxonomist: 16%

^Microzooplankton: 80.0% (± 

9.4%) PS, n=4 and 42.4% (± 

32.2%) RPD, n=4 

Macrozooplankton: 97.3% (± 

2.5%) PS, n=5 and 4.0% (± 

3.9%) RPD, n=5 

Representativeness 

Pre-treatment/control and 

post-treatment/treatment 

samples are handled and 

analyzed in the same manner. 

Not Applicable 

– Qualitative. 

All pre-treatment and post-treatment zooplankton 

samples collected on intake, as well as all control and 

treatment discharge samples, were collected, handled, 

and analyzed in the same manner (using the appropriate 

GSI SOPs). 

Comparability 

Routine procedures are 

conducted according to 

appropriate SOPs to ensure 

consistency between tests. 

Not Applicable 

– Qualitative. 

The following GSI SOP was used for all zooplankton 

sample analyses conducted during the test trials: 

- GSI/SOP/LB/RA/SA/2 (DRAFT) – Procedure for 

Zooplankton Sample Analysis 

Completeness 

Number of valid data obtained 

from the performance 

measurement system vs. 

number performance 

measurement samples 

analyzed. 

Performance is measured by 

percent completeness (%C). 

>90% C. 

^Microzooplankton: 2/8 performance measurements 

met DQO = 25% C

Macrozooplankton: 9/10 performance measurements 

met DQO = 90% C

^Zooplankton analysis parameter having performance measurement result that did not meet data quality 

objective specified in GSI QAPP (2009). 
GSI/LB/F/A/1 

March 15, 2010 

Page 56 of 58 

Table 5. Microbial Analyses. 

Data Quality 

Indicator 

Evaluation Process/ 

Performance 

Measurement 

Data 

Quality 

Objective 

Performance Measurement Result 

Precision 

Samples (10%) are 

analyzed in duplicate – 

with performance 

measured by average 

relative percent difference 

(RPD) of all duplicate 

analyses performed during 

test trials. 

<20% 

average (± 

SEM) RPD. 

Percentage of Microbial 

Samples Analyzed in 

Duplicate: 20%

^E. coli: 23.25% (±9.31%) RPD, n=12

^Total Coliforms: 36.36% (±14.99) RPD, n=12

^Enterococcus spp.: 26.96% (±10.27%) RPD, 

n=10

^Heterotrophic: 53.93% (±24.96%) RPD, n=10

^Vibrio spp.: 31.08% (±8.41%) RPD, n=16

Bias 

Samples (10%) are counted 

by two separate analysts – 

with performance measured 

by average relative percent 

difference (RPD) for all 

second counts performed 

during test trials.

<20% 

average (± 

SEM) RPD. 

Percentage of Microbial 

Samples Counted by a 

Second Analyst: 40%

(Total Coliforms); 30%

(E. coli, Enterococcus 

spp., and Heterotrophic 

Bacteria); and 10%

(Vibrio spp.)

E. coli: 0.00% (±0.00%) RPD, n=24

Total Coliforms: 4.17% (±4.17) RPD, n=32

Enterococcus spp.: 13.02% (±12.65%) RPD, n=24

Heterotrophic: 0.84% (±0.58%) RPD, n=24

^Vibrio spp.: 43.06% RPD, n=15

Representativeness 

Pre-treatment/Control and 

post-treatment/treatment 

samples are handled and 

analyzed in the same 

manner. 

Not 

Applicable 

– 

Qualitative. 

All pre-treatment and post-treatment water quality intake samples, as well 

as all control and treatment discharge samples, were collected, handled, 

and analyzed in the same manner (using the appropriate GSI SOPs). 

Comparability 

Routine procedures are 

conducted according to 

appropriate SOPs to ensure 

consistency between tests. 

Not 

Applicable 

– 

Qualitative. 

The following GSI SOPs were used for all microbial analyses conducted 

during the test trials: 

-GSI/SOP/BS/RA/MA/1 – Procedure for Quantifying Heterotrophic Plate 

Counts (HPCs) using IDEXX’s SimPlate® for HPC Method

-GSI/SOP/BS/RA/MA/3 – Procedure for the Detection and Enumeration of 

Enterococcus using Enterolert® 

-GSI/SOP/BS/RA/MA/4 – Procedure for the Detection and Enumeration of 

Total Coliforms and E. coli using IDEXX’s Colilert® 

-GSI/SOP/RDTE/SA/M/3– Procedure for Colony Blot Preparation for the 

Enumeration of Culturable Vibrio cholerae

Completeness 

Number of valid data 

obtained from the 

performance measurement 

system vs. number of 

performance 

measurements collected. 

Performance is measured 

by percent completeness 

(%C). 

>90% C. 

^E. coli: 31/36 performance measurements met DQO = 86% C

^Total Coliforms: 36/44 performance measurements met DQO = 82% C

^Enterococcus spp.: 27/34 performance measurements met DQO = 79% C

^Heterotrophic: 27/34 performance measurements met DQO = 79% C

^Vibrio spp.: 15/31 performance measurements met DQO = 48% C

Sensitivity 

The limit of detection (LOD) 

for the analytical method 

used is reported. 

Dependent 

upon the 

analytical 

technique 

used. 

E. coli LOD: <1 MPN/100 mL 

Total Coliforms LOD: <1 MPN/100 mL 

Enterococcus spp. LOD: <1 MPN/100 mL 

Heterotrophic LOD: <2 MPN/1 mL 

Vibrio spp. LOD: 0 CFU/100 mL 

^Microbial analysis having performance measurement result that did not meet data quality objective specified in 

GSI QAPP (2009). 
GSI/LB/F/A/1 

March 15, 2010 

Page 57 of 58 

Table 6. Whole Effluent Toxicity (WET) Tests. 

Data Quality 

Indicator 

Evaluation Process/ 

Performance 

Measurement 

Data Quality 

Objective Performance Measurement Result 

Bias 

Experiment Bias:

Monthly reference 

toxicant tests are 

conducted on test 

organisms. Performance 

is measured by 

sensitivity of the test 

organisms relative to 

historical values. 

LC50 value 

within 2 

standard 

deviations of 

the historical 

LC50 average 

C. dubia: LC50 values from reference toxicant tests performed during WET 

Testing were within 2 SD of the historical average (501.5 and 498.4 mg/L 

KCl), n=2. 

P. promelas: LC50 value from reference toxicant test performed 9/2009 was 

within 2 SD of the historical average (6.45 g/L NaCl), n=1. 

S. capricornutum: No data, reference toxicant test not conducted on this 

test organism. 

Operator Bias:

Experimental units (10%) 

are counted by two 

separate analysts – with 

performance measured 

by average relative 

percent difference (RPD)

of the number of live 

test organisms counted 

for all second analyses. 

<10% average 

RPD (±SD) 

C. dubia: 39% of 

experimental units 

counted by a second 

analyst 

C. dubia: 0.1% (± 0.23%) RPD, n=1055 

P. promelas: 35% of 

experimental units 

counted by a second 

analyst 

P. promelas: 0.1% (± 0.1%) RPD, n=442 

S. capricornutum: 0%

of experimental units 

counted by a second 

analyst. 

S. capricornutum: No data, RPD could not be 

determined 

Representativeness 

Control groups 

(reference and dilution 

control) and treatment 

groups are handled and 

analyzed in the same 

manner. 

Not 

Applicable – 

Qualitative. 

All control groups (reference and dilution controls) and all treatment groups 

were set up, handled, and analyzed in the same manner (using the 

appropriate GSI SOPs). 

Comparability 

Routine procedures are 

conducted according to 

appropriate SOPs to 

ensure consistency 

between tests. 

Not 

Applicable – 

Qualitative. 

The following GSI SOPs were used for all WET Testing conducted during the 

test trials: 

-GSI/SOP/BS/RA/WET/1 – Procedure for Assessing Chronic Residual 

Toxicity of a Ballast Treatment System to Ceriodaphnia dubia

-GSI/SOP/BS/RA/WET/2 – Procedure for Assessing Chronic Residual 

Toxicity of a Ballast Treatment System to the Fathead Minnow (Pimephales 

promelas) 

-GSI/SOP/BS/RA/WET/3 (DRAFT) – Procedure for Assessing Chronic 

Residual Toxicity of a Ballast Treatment System to the Green Alga 

(Selenastrum capricornutum) 

Completeness 

Number of valid data 

obtained from the 

performance 

measurement system vs. 

number of performance 

measurements collected. 

Performance is 

measured by percent 

completeness (%C). 

>90% C. 

Ceriodaphnia dubia: 1075/1075 performance measurements met DQO = 

100% C

Pimephales promelas: 443/443 performance measurements met DQO = 

100% C

Selenastrum capricornutum: No data, %C could not be determined.
GSI/LB/F/A/1 

March 15, 2010 

Page 58 of 58 

APPENDIX 5. OPERATIONAL DATA SUMMARY 

Trial A Trial B Trial C Trial D Trial E 

Main System 

Fill 

Total Volume 

Control Retention Tank m^3 192 192 193 193 193

Treatment Retention Tank m^3 193 193 193 193 192

Avg Flow 

Control Track m^3/hr 199 200 199 199 200

Treatment Track m^3/hr 199 198 199 199 198

Avg System 

Pressure Both Tracks kPa 132 130 132 129 130

Discharge 

Total Volume 

Control Retention Tank m^3 180 181 181 181 182

Treatment Retention Tank m^3 182 179 184 179 182

Avg Flow 

Control Track m^3/hr 200 196 196 199 199

Treatment Track m^3/hr 200 198 197 199 199

Avg System 

Pressure 

Control Track kPa 135 130 128 125 123

Treatment Track kPa 125 124 128 127 130

Avg Values Calculated from the point of the by-pass valve shut until the pump is off.

Trial A Trial B Trial C Trial D Trial E 

Sampling System 

Fill 

Control Sample Tub 1 m^3 2.09 2.09 2.09 2.08 2.09

Sample Tub 2 m^3 2.08 2.08 2.08 2.07 2.08

Treatment Sample Tub 4 m^3 2.11 2.09 2.09 2.08 2.11

Sample Tub 5 m^3 2.11 2.09 2.09 2.08 2.11

Post 

Treatment Sample Tub 6 m^3 2.09 2.09 2.08 2.07 2.09

Drain 

Control 

Sample Tub 1 m^3 3.78 2.07 2.06 2.04 2.05

Sample Tub 2 m^3 3.77 2.07 2.06 2.03 2.03

Sample Tub 3 m^3 2.07 2.06 2.03 2.04

Treatment 

Sample Tub 4 m^3 3.88 2.02 2.06 2.02 2.04

Sample Tub 5 m^3 3.87 2.02 2.06 2.01 2.04

Sample Tub 6 m^3 2.01 2.06 2.01 2.04

Avg Values calculated from the measurable start of flow to the end of measurable flow