I:\MEPC\56\2-1.doc For reasons of economy, this document is pr inted in a limited number. Delegates are kindly asked to bring their copies to mee tings and not to request additional copies. INTERNATIONAL MARITIME ORGANIZATION

IMO 

# E

MARINE ENVIRONMENT PROTECTION

COMMITTEE
56th session Agenda item 2 MEPC 56/2/1

15 December 2006 Original: ENGLISH

HARMFUL AQUATIC ORGANI SMS IN BALLAST WATER

Application for Final Approval of a ballast water management system using Active Substances

Submitted by Norway

SUMMARY

Executive summary: This document contains an application for Final Approval of a ballast water management system under the Procedure for approval of ballast water management systems that make use of Active Substances (G9) adopted by resolution MEPC.126(53)

Action to be taken: Paragraph 6

Related documents: BWM/CONF/36, MEPC 53/24/Add.1 and BWM.2/Circ.6

Introduction

1 Regulation D-3.2 of the International Conve ntion for the Control and Management of Ships’ Ballast Water and Sediments stipulates th at ballast water management systems that make use of Active Substances to comply with the Convention shall be appr oved by the Organization.

2 The Procedure for approval of ballast water management systems that make use of Active Substances (G9) contains the information that should be included into a proposal for approval (MEPC 53/24/Add.1, annex 4, paragraph 4.2.1) and provisions for risk characterization and analysis (MEPC 53/24/Add.1, annex 4, paragra ph 5.3). According to section 6 of the Procedure (G9) the Organization should evaluate the informati on provided in the application.

3 According to BWM.2/Circ.6, a document on th e proposal for approval containing all the

non-confidential data related to the respective Active Substances shall be submitted to MEPC 56.

4 The competent authority in Norway has verifi ed the application dossier and believes it satisfy the data requirements of the Proce dure (G9) adopted by resolution MEPC.126(53).

5 Norway therefore submits the non-confidentia l part of the manufacturer’s application

dossier in the annex of this document to th e Organization for evaluation according to the Procedure (G9). The complete dossier will be made available to the experts of the

GESAMP-BWWG with the understanding of confid ential treatment.

Action requested of the Committee

6 The Committee is invited to consider the proposal for approval and to decide as appropriate.

***MEPC 56/2/1

I:\MEPC\56\2-1.doc ANNEX

PROPOSAL FOR APPROVAL ALFA LAVAL BALLAST WATER MANAGEMENT

SYSTEM - PUREBALLAST

This report is structured in accordance with th e headlines in the “Procedure for approval of Ballast Water Management systems that make use of Active Substances (G9)”.

The documentation includes:

• General report with process description and responses according to Procedure (G9).

• Reference list for process description.

Attachments:

1. Lloyds approval of full scale prototype system onboard M/V Don Quijote.

2. Extraction of QAPP for Full scale Type Appr oval testing according to Guidelines for approval of ballast water management sy stems (G8) and the Procedures (G9).

3. Assessment report of Full scale toxicity testing

− Det Norske Veritas.

4. Full scale toxicity testing.

Reference dossier (additions to Reference dossi er sent together with “Proposal for Basic Approval)”

Summary:

The type approval test performed, according to Gu idelines (G8) and Procedure (G9) for this report, shows that PureBallast meets the D-2 di scharge standard and has no significant toxic

effects or residuals.MEPC 56/2/1

ANNEX Page 2

I:\MEPC\56\2-1.doc 1 INTRODUCTION

Working Principles of the Alfa Laval Balla st Water Management System (PureBallast)

The ballast water is treated at intake and once again at discharge. The treatment on intake ensures that a minimal amount of viable organism s enter into the ballast water tanks and reduces sediment built-up in the tanks, whic h is a potential area for survival of organisms. The water is treated again at discharge to ensure that any pot ential re-growth of organisms in the ballast water tanks is prohibited.

Description of the PureBallast system and its components

The PureBallast product is a complete syst em, composing of three main functions:

• Filter (50 μm) An automatic back flushing filter for removal of larger particles and organisms as well as to reduce the level of sediments in the ballast water tanks.

• Wallenius AOT (Advanced Oxidation Technology) Unit.

A patented system product using naturally o ccurring photo-processes in the surface layer of sea-water. The Wallenius AOT process acts on two levels: in-situ formation of reactive free radicals, and direct photo-effects. These two levels synergistically inactivate micro-organisms. A special UV light wavelength spectrum is used in combination with a catalyst which is based on TiO 2 . The TiO 2 acts as a true catalyst and is not consumed in the process. No substances are added to the process.

• Control and Auxiliary Equipment including sampling points.

The control system and transformer control the system and supply a ll items with power. The auxiliary equipment controls the flow and measure different alarm levels. The system also composes a CIP unit which is an auto matic service device th at cleans the quartz sleeves covering the UV-lamps after each Ballast or Deballast sequence. This operation is done to avoid scaling from seawater contam inants which would otherwise reduce the efficiency of the Wallenius AOT Module.

This application will focus on th e Wallenius AOT treatment unit. The filter and other components are not considered to be relevant for Active Substance approval.

Control and Auxiliary Equipment including sampling points

• Control System An overall control system th at controls the process, log all data and give alarms.

• Transformer Supplies all units with po wer independent of available power properties on ship.

• Sampling points Allows for accurate sampling befo re and after treatment system.

• CIP (Cleaning In Place) Support equipment for automatic cleaning of th e quartz sleeves protecting the UV-lamps.MEPC 56/2/1

ANNEX

Page 3

I:\MEPC\56\2-1.doc PureBallast Test System Description

Description of pilot scale test syst em used for Basic Approval testing

Pilot scale testing was done prior to the start of these full scale tests.

The first test was made in April 2006, used for test program for Basic A pproval application. The tested system was carefully scal ed and corresponds to the full scale unit, but used a higher input power, relative to the flow.

The following properties are relevant for the pi lot scale test system used in April 2006:

Specified Flow rate 25 m 3 /h Power consumption 10 kW Number of lamps 10

A second test was made in June 2006, used for pre- test prior to full scale testing. The tested system is carefully scaled and corresponds to the full scale unit.

The following properties are relevant for the pi lot scale test system used in June 2006:

Specified Flow rate 25 m 3 /h Power consumption 6 kW Number of lamps 10

Description of the full scale test sy stem used for Type Approval testing

The full scale test system is designed for the full scale land based testing including the “Final Approval”

− test program as specified in the at tached QAPP-extraction, see appendix 1.

The following properties are relevant for the full scale test system:

Specified Flow rate 250 m 3 /h Power consumption 60 kW Number of lamps 20 (2x10) Treatments Ballast + Deballast (Two treatments)

Treated volume >200 m 3

The toxicity testing in this application is pe rformed during the type ap proval land based testing performed completely according to G8 Guideline, see appendix 3. The deta iled procedures and a complete test set up are included in the QA PP from NIVA (Norwegian Institute for Water Research), see appendix 2.

A complete treatment cycle is two stage treatments which correspond to Ballast as well as Deballast. A complete treatment cycle is used fo r all toxicity and chemical water property tests which is presented in this application.MEPC 56/2/1

ANNEX Page 4

I:\MEPC\56\2-1.doc 2 DEFINITIONS

Interpretation for the PureBallast system (focus on Wallenius AOT unit)

“Ballast Water Discharge” is the treated ballast water that has passed the system twice, first during Ballast and then after Deballast.

The term “Preparation” refers to the extremely short lived free radicals generated in treated ballast water from the Wallenius AOT process.

Due to the short life span of the chemistry in th e treatment process, all “Relevant Chemicals” as defined by IMO is the potential transformation or reaction products in the treated water.

The above interpretation of defi nitions indicates that the term s “Active Substance”, “Ballast Water Discharge”, “Preparation” and “Relevant Chem icals” all refers to the treated ballast water from the Wallenius AOT process. This means that it is the treated water that is considered in this application and thus tested regarding toxicity and content of chemicals.

3 PRINCIPLES

No comment.

4 GENERAL REQUIREMENTS

4.1 Identification

The following section describes the chemical f eatures and discharge characteristics of the Wallenius AOT process.

Introduction

The process description pres ented is based upon the current scientific knowledge about photo-catalysis, free radical a nd marine chemistry cited in the references. Although the Wallenius AOT-process generates dichloridyl and di bromidyl radicals in sea water, it does not involve any formation of elemental chlorine (Cl2 ) or bromine (Br2). Elemental chlorine and bromine react with organic materials by formation of halogenated products, which can give rise to environmental stress. This chemistry is not operative in the Wallenius AOT-process, and consequently, neither is the associated environmental concern. It should be point ed out that the radicals produced in the AOT-treatment unit are not different from those naturally formed by sun light in the surface layer of s eawater. Any potential reaction pr oduct that may be formed have been proposed and measured to support the chemical process description. In fact, when chemical changes were observed they indicated a reductio n, e.g., the content of chloroform was reduced. Most significantly, all biological tests performed have shown that the water treated by the Wallenius AOT-process does not display any adverse effect.

Discharge characteristics

The basic process operates without addition of chem icals using the synergistic effects of in situ produced free radicals and UV-photons to inactivate micro-organisms. Per se , TiO 2 is non-toxic as shown by its use in re-constructive surgery to obtain bone-compatible links for Ti-implantsMEPC 56/2/1

ANNEX

Page 5

I:\MEPC\56\2-1.doc such as teeth and hip-joints (Pan et al. 1996, Diebold 2002). TiO 2 is also a widely used component in paints and cosmetic products. The action of reactive free radicals is of temporal

nature , i.e. these species are so short-lived (micro-m illi second time-scale) that they are not

observable outside the AOT-treatment unit (Baxendale and Busi 1982, Buxton et al. 1988,
NDRL/NIST Solution Ki netics Database 2005).

The only environmental concern of the PureBallast treatment system might be attributed to the reaction products. To elucidate whether any envir onmental effects are to be expected, a series of chemical analysis and ecotoxicological tests of possible formation of t oxic reaction products and their potential effects is relevant. Of primary inte rest would be the slight possibility of formation of halogenated organics or toxic oxi dation products (for example bromate).

A problem related to Ozone treatment of sea wa ter is a significant formation of bromate, a suspected carcinogen. This reaction may proceed by way of an oxygen atom transfer mechanism. In the photo-catalytic process formation of bromate, BrO 3 - , is unlikely to occur, since this would require multiple one-electron oxidation steps. Moreover, the utilization of photocatalysis is one of the attract ive methods for the removal of BrO 3 - . The reduction of BrO 3 -
on a TiO 2 photocatalyst under UV irradi ation is a well-known reac tion, and bromate has even been used as an additive to improve the oxidation efficiency of TiO 2 photocatalysts (Oosawa and Gretzel 1988, Al-Ekabi et al. 1993, Mills et al . 1996, Lindner et al. 1997, Noguchi et al. 2003)

It should be pointed out that the radicals produc ed in the AOT-treatment unit are not different from those naturally formed by sun light in th e surface layer of seaw ater. In this case, photo-cleavage of hydrogen peroxide and/or photo- oxidation of nitrate/nitrite ions may be the

source of radical formation.

Therefore, treated ballast water is tested for toxicity and chemical properties, specifically for:

• Chloro-organic compounds

• Bromo-organic compounds

• A general toxicity anal ysis (aggregate acute and chronic toxicity)

• Bromate formation

• THM formation (Tri halo methane)

• Octanol/Water partition coefficient

4.2 Data-set for Active Substances and Preparations

The potential toxic effects of the treatment were tested in a set of bioassays; see Appendix 4 for the tests review and test reports.

4.2.1 A proposal for approval should include inform ation on the properties or actions of the Preparation including any of its components as follows:

.1 Data on effects on aquatic plants, inverteb rates, fish, and other biota, including sensitive and representative organisms:

• acute aquatic toxicity;

No significant acute toxicity was observed. See report in appendix 4,

section 3.7.MEPC 56/2/1

ANNEX Page 6

I:\MEPC\56\2-1.doc • chronic aquatic toxicity;

No significant chronic toxicity wa s observed. See report in appendix 4,

section 3.7.

• endocrine disruption;

Not tested specifically, however juve nile development and reproduction on

vertebrate as well invertebrate test or ganism were not found to be affected (see appendix 4, section 3.7). More over, no significant chemicals were identified as potential endocrine af fecting substances were found in the treated water (see appendi x 4, section 3.1). This suggests endocrine disruption as being unlikely to occur.

• sediment toxicity;

As showed in the chemical and toxic ity testing there will be no significant residuals resulting from the treatment and therefore no particle flux to the sediment. Sediment toxicity is therefore not expected.

• bioavailability/biomagni fication/bioconcentration;

Octanol/water partitioning does not suggest bioaccumulation/biomagnification/bi oconcentration to occur, see appendix 4, section 3.1.

• food web/population effects;

Since the full strength effluent (100%) did not have any deleterious effects on any of the test species, the effluent would not be expected to be toxic to aquatic organisms and have negative effects on ecological communities at environmentally relevant concentrations.

.2 Data on mammalian toxicity: Not relevant according to the results above.

• acute toxicity; • effects on skin and eye; • chronic and long-term toxicity; • developmental and reproductive toxicity; • carcinogenicity; and • mutagenicity.

.3 Data on environmental fate and effect under aerobic and anaerobic conditions:

• potential physical effects on wildlife and benthic habitats;

Not tested. Treatment does no affect phys ical properties of the water, see appendix 4, section 3.1. No potential phys ical effects could be expected.MEPC 56/2/1

ANNEX

Page 7

I:\MEPC\56\2-1.doc • potential residues in seafood;

Since bioaccumulation effects are not expected, no residue s resulting from the treatment are likely to occur.

• any known interactive effects.

No interactive effects are expected.

• Modes of degradati on (biotic; abiotic);

The primary active species are of sufficient reactivity to either react or

terminate with each other well with in the milli-second to second time scales and will thus be consumed before they can leave the reactor. The primary residues will therefore be r eaction products stemming from the oxidation (major) or reduction (minor) caused by the free radicals initially

formed. As these are formed an alogously to natural decomposition processes they can be expected to be naturally degradable by natural biotic or abiotic paths.

• Bioaccumulation, partition coeffici ent, octanol/water coefficient;

The potential for bioaccumulation of the treated water is assessed by measurement of possible changes in th e octanol/water partition values of treated water. No changes have been found, see test results for chemical properties in appendi x 4, section 3.1.

• Persistence and identification of the ma in metabolites in the relevant media (ballast water, marine and fresh waters);

As no oxidation products were identified and no significant measurable quantities of those were found, the pers istence assessment is not relevant.

• Reaction with organic matter;

The process generates free radicals of sufficient reactivity able to react with most organic compounds. The major modes of reaction will be one-electron transfer (Br 2 -· , CO 3 -· , Cl 2 -· radicals), hydrogen abstraction (hydroxyl, CO 3 -· radicals) or addition r eactions (hydroxyl radicals).

All these radicals are natura lly formed in the surfaces of (sea) water so any reaction products can be expected to be of a character similar to those of “natural” decomposition in water. It should be noted that the reactive free radicals will react indiscriminately fo rming an array of different reaction products by various reaction paths, ma king it difficult to predict specific products. Generally these kinds of processes are discussed for environmental applications to reduce the amount of hazardous material in

water streams.MEPC 56/2/1

ANNEX Page 8

I:\MEPC\56\2-1.doc .4 Physical and chemical properties for the Active Substances a nd Preparations and the treated ballast water, if applicable:

This technology is a physical-chemical pr ocess and does not i nvolve the addition of any substances. The in-situ generated free radicals have life times (half life) of

micro or milli seconds and are c onsumed inside the reactor.

• melting point;

Not applicable.

• boiling point;

Not applicable.

• flammability;

Not applicable

. • density (relative density);

Not applicable.

• Vapour pressure, vapour density;

Not applicable.

• water solubility/dissociation constant (pKa);

The process may increase the amount of oxidized groups in the dissolved organic matter, leading to a decrease in COD, i.e. increase in oxidized groups like: acidic, alcoho lic, aldehyde and ketostru ctures. This may alter the pKa or buffer properties of the treated water.

• oxidation/reduction potential;

No change of the oxidation/reduction pot ential due to the AOT process is expected, see appendix 4, section 3.1.

• corrosivity to the materials or equipment of normal ship construction;

No change of corrosion potential due to the AOT process is expected.

• autoignition temperature;

Not applicable.

• other known relevant physic al or chemical hazards.

None at this date.MEPC 56/2/1

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Page 9

I:\MEPC\56\2-1.doc .5 Analytical methods at environm entally relevant concentrations.

4.2.2 A proposal for approval should include the above data set eith er for the Preparation or for each component separately, and a list of the name and relative quantities (in volumetric percentages) of the component s should be also attached. As described in section 8.1, all proprietary data should be treated as confidential.

Not applicable.

4.2.3 The tests for Active Substances and Prepara tions should be carried out in accordance with internationally recognized guidelines.

4.2.4 The testing process should contain a rigor ous quality control/quality assurance program consisting of:

All analyses are done according to standard me thods and toxicity tes ting by NIVA (Norwegian Institute of Water Research) according to define d QAPP (QMP included), see an extraction of used QAPP (QMP included) in appendix 2. The pe rformed tests are supervised by Det Norske Veritas, see appendix 3.

.1 Both a Quality Management Plan (QMP) and a Quality Assurance Project Plan (QAPP). Guidance on preparation of th ese plans, along with other guidance documents and other general quality control information are available for download from the International Orga nization for Standardization (ISO) ( www.iso.org ).

See an extraction of used QAPP with QMP included in appendix 2.

.2 The QMP addresses the quality control ma nagement structure and policies of the Test Organization (inclu ding subcontractors and outside laboratories).

.3 The QAPP is a project specific technical document reflecting the specifics of the system to be tested, the test facility, and other conditions affecting the actual design and implementation of the required experiments.

4.2.5 Dossiers already used for registration of ch emicals can be submitted by the applicant to satisfy the required data needed for the eval uation of Active Substances and Preparations according to this procedure.

4.2.6 The proposal should describe the manner of application of the Preparation for Ballast Water Management, including requir ed dosage and retention time.

4.2.7 A proposal for approval should include (Mat erial) Safety Data Sheets ((M)SDS).

4.3 Assessment report

4.3.1 A proposal for approval should include an assessment report. The assessment report should address the quality of the te st reports, the risk characteri zation and a consid eration of the uncertainty associated with the assessment.MEPC 56/2/1

ANNEX Page 10

I:\MEPC\56\2-1.doc The test results found no significan t levels of potentially toxic s ubstances and no significant acute

or chronic effects of the full st rength effluent on aquatic organi sms. Therefore, no apparent environmental risks could be identified. See appendix 4.

All analyses are done according to standard methods and toxic ity testing is performed under supervision by Det Norske Veritas. Their asse ssment report is attached as appendix 3.

5 RISK CHARACTERIZATION

5.1 Screening for persistency, bioaccumulation and toxicity

5.1.1 An assessment on the intrinsic properties of the Active Substa nce and/or Preparation such as persistency, bioaccumulation and toxicity shou ld be conducted (see Table 1 in section 6).

.1 Persistence tests:

Persistence should preferably be assess ed in simulation test systems that determine the half-life under relevant conditions. Biodegradation screening tests may be used to show that the substa nces are readily biodegradable. The

determination of the half-life should include assessment of relevant chemicals.

See description in section 4.1, headline “ Radicals formed and their expected lifetime”.

.2 Bioaccumulation tests:

The assessment of the (potential fo r) bioaccumulation should use measured bioconcentration factors in marine (or fr eshwater) organisms. Where these tests are not applicable, or if logPow <3, Bi o Concentration Factor (BCF) values may be estimated using (Quantitative) Structure-Activity Relationship ((Q)SAR)

models.

The potential for bioaccumulation of the treated water are assessed by measurement of possible changes in the oc tanol/water partition values of treated water. No changes has been found, see te st results for chemi cal properties in appendix 4. The octanol/water part itioning values do not suggest bioaccumulation/biomagnification/bioconcentration to occur.

.3 Toxicity tests:

Acute and/or chronic ecotoxicity data, id eally covering the sensitive life stages, should in principle be used for the assessment of the to xicity criterion.

Acute and chronic tests covering survival on three trophic levels, including marine algae, marine crustaceans and juveniles fish indicated no significant acute or chronic toxicity to any of these endpoints in any test organisms at any effluent

concentrations as tested. Since the full st rength effluent (100%) did not have any deleterious effects on any of the test species , the effluent would not be expected to be toxic to aquatic organisms and have negative effects on ecological communities at environmentally relevant concentrations.MEPC 56/2/1

ANNEX Page 11

I:\MEPC\56\2-1.doc 5.2 Toxicity testing of the treated Ballast Water

5.2.1 Toxicity testing is necessary for the Active S ubstance, or Preparati ons (see sections 4.2.1 and 5.3) and the treated Ballast Water Discharge as covered in this section. The advantage of conducting toxicity testing on the Ballast Water Disc harge is that it integrates and addresses the potential for interactions of the Active Subs tances and Preparations with the possible by-products.

.1 For the basic approval process, the disc harge testing should be performed in a laboratory using techniques and equipmen t to simulate Ballast Water Discharge following treatment by the Preparation.

.2 For final approval, the discharge testi ng should be performe d as part of the land-based type approval process using th e treated ballast water discharge.

5.2.2 The applicant should provide both acute and chr onic toxicity test data using standardized test procedures to determine the toxicity of the Preparation and Relevant Chemicals as used in conjunction with the Ballast Wa ter Management System. This testing approach should be performed on the treated Ballast Water Discharg e, as the Ballast Water Management system could either mitigate or enhance the adverse effe cts of the Preparation or Relevant Chemicals.

5.2.3 The discharge toxicity tests should be conducted on samples drawn from the land-based test set-up, which would be representative of th e discharge from the Ballast Water Management
system.

5.2.4 These toxicity tests should include chronic test methods with multiple test species (a fish, an invertebrate and a plant) that address the sensi tive life-stage. The prefer ence is to include both a sub-lethal endpoint (growth) and a survival endpoint. Either fres hwater or marine test methods

should be tested.

5.2.5 The test results to be provided include : acute 24-hour, 48-hour, 72-hour, and 96-hour
Lethal Concentration at which x % of the test or ganisms die (LCx), No Observed Adverse Effect Concentrations (NOAECs), chronic No Observed Effect Concentration (NOEC) and/or Effect Concentration at which x % of test organisms s how effect (ECx), as appropriate based on the experimental design.

5.2.6 A dilution series including a 100% ballast wa ter discharge would be tested to determine the no adverse effect level using the statistical endpoints (NOEC or ECx). An initial analysis could use a conservative approach where th e dilution capacity woul d not be taken into consideration (no modeling or plumes analysis would be used). The rationale for taking a conservative approach is that there could be mu ltiple discharges into one location (even though this is not necessarily the case).

The actual NOEC were not measurable, since no si gnificant toxic effects has been detected even using 100% treated ballast water.

5.2.7 The acute and chronic toxicity test data in conjunction with the information in

Section 4.2.1 should be used to determine the ho lding time necessary to achieve the no adverse effect concentration upon discha rge. Knowing the ha lf-life (days), decay rate, dosage rate, volume of system and toxicity tests with time seri es, then a computational model can be used to

determine the amount of time needed to hold th e treated ballast water before discharge.MEPC 56/2/1

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I:\MEPC\56\2-1.doc 5.3 Risk characterization and analysis

5.3.1 For the basic approval process, fate and effect testing should be performed in the laboratory with Active Substances and Preparations. This section lists information that could be useful for a preliminar y risk characterization.

5.3.2 Both the Active Substance or Preparation as well as the treated Ballast Water Discharge should be subject to toxi city testing in order to protect the receiving environment from toxic effects due to discharges.

These are all the same substance as di scussed in Interpretation of Definition.

5.3.3 The reaction with organic matter of Active S ubstances and Preparations that produce free radicals, should be addressed qualitatively so as to identify products of concern to the

environment.

See description in section 4.1, headline “ Radicals formed and their expected lifetime”.

5.3.4 The rate of abiotic and biotic degradati on of the Active Substances and Preparations

under aerobic and anaerobic conditions should be assess ed, resulting in th e identification of

relevant metabolites in the re levant media (ballast water, marine and fresh waters).

This process does not involve the addition of chemicals.

5.3.5 The rate of abiotic and biotic degradati on of the Active Substances and Preparations

under aerobic and anaerobic conditions should be a ssessed, resulting in th e characterization of the persistence of the Active Substances, Prepar ations and Relevant Chemicals in terms of degradation rates under specified co nditions (e.g., pH, redox, temperature).

See description in section 4.1, headline “ Radicals formed and their expected lifetime”.

5.3.6 The partition coefficients (solids-water part ition coefficient (Kd) and/or organic carbon normalized distribution coefficient (Koc)) of the Active Substances, Preparations and Relevant

Chemicals should be determined.

This process does not involve the addition of chem icals and therefore this is not applicable.

5.3.7 For Active Substances and Preparations, th e potential for bioaccumulation should be

assessed in marine or freshwater organisms (fish or bivalves) if the logarithm octanol/water partition coefficient (logPow) is >3.

This process does not involve the addition of chemicals. The pot ential for bioaccumulation of the treated water was assessed by measurement of possible (if any) change s in the octanol/water partition values of treated water. This analysis showed that no changes occurred in the log Pow

before and after treatment.

5.3.8 Based on the information on fate and behavior of Active Substances and Preparations, the discharge concentrations at selected time intervals should be predicted.MEPC 56/2/1

ANNEX Page 13

I:\MEPC\56\2-1.doc As showed in the chemical and toxicity testin g there will be no significan t residuals or effects directly after the Wallenius AOT unit and theref ore discharge concentrations at different time

intervals is not relevant.

5.3.9 The effect assessment of the Active Substanc es, Preparations and Re levant Chemicals is initially based on a dataset of acu te and/or chronic ecot oxicity data for aquatic organisms, being primary producers (algae or sea grasses), cons umers (crustaceans), predators (fish), and should include secondary poisoning to mammalian and av ian top-predators, as well as data for sediment species.

The assessment was based on acute and chronic toxicity to primar y producers (alg ae), herbivore consumers (rotifers, crustaceans) and juvenile fish.

5.3.10 An assessment of secondary poisoning is redundant if the substance of concern demonstrates a lack of bioaccumulation potenti al (e.g., BCF <500 L/kg wet weight for the whole organism at 6% fat).

Secondary poisoning testing is not relevant as octanol/water partitioning does not suggest

bioaccumulation to occur.

5.3.11 An assessment of sediment species is redund ant if the potential of the substance of concern to partition into the sedi ment is low (e.g., Koc <500 L/kg).

The partition into the sediment cannot exceed th e indicated value; the assessment is therefore

redundant.

5.3.12 The effect assessment of the Active Substances, Preparations and Relevant Chemicals should include a screening on carcinogenic, muta genic and endocrine di sruptive properties. If the screening results give rise to concerns, this shoul d give rise to a further effect assessment.

As no carcinogenic, mutagenic and endocrine disr uptive substances were found in the analyzed water after the treatment, the assessment is not relevant.

5.3.15 An analytical method suitable for monitori ng Active Substances a nd Preparations in ballast water discharges should be available.

As showed in the chemical and toxicity testin g there will be no significan t residuals or effects directly after the Wallenius AOT unit and ther efore monitoring of “A ctive Substances” or “Preparations” in the discharge is not relevant.

6.3 Ship and personnel safety

6.3.1 In order to protect the ship and personnel sa fety the technical group should evaluate the physical and chemical hazards (see paragraph 4.2.1.4) to ensure that potential hazardous properties of the Active Substances, Preparations or Relevant Chemicals formed in the treated

ballast water should not create any unreasonable risk to the ship and personnel. Proposed procedures for the use and tec hnical equipment introduced needs to be taken into account.

The only potential risk present in the PureBallast system identified, which is related to the Wallenius AOT process, is the UV-irradiati on from the UV-lamps and overheating in the Wallenius AOT treatment unit. If the flow is cut off the lamps will be automatically shut-off soMEPC 56/2/1

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I:\MEPC\56\2-1.doc there is no risk of overheating. Several independent actions ensure that th e shut-off is made. The lamps are completely enclosed in the Wallenius AOT chambers and can not be operated outside

the chamber. There is therefore no risk that any personnel are exposed to the UV-irradiation.

The free radicals will not be able to leave the Wallenius AOT tr eatment unit, as described in section 4.1, headline “ Radicals formed and th eir expected lifetime”.

A full scale prototype system has been placed onboard a car carrier. The system has been evaluated by Lloyd’s classification society and has been approved w ith regards to ship and crew safety, see appendix 1.

6.3.2 For the protection of personnel involved in the handling and storage of the Active Substances and Preparations, the proposal should incl ude relevant ((M)SDS). The Organization should evaluate (M)SDS, mammalian toxicity data and chemical properties hazards (see paragraphs 4.2.1.2 and 4.2.1.4) and ensure that potential hazardous pr operties of the Active Substances, Preparations or Relevant Chemical s should not create any unreasonable risk to the ship or personnel. This evaluation should take into account the different circumstances that a ship or personnel may face in its trad e (e.g., ice, tropical, humidity, etc.).

Not applicable.

6.4 Environmental protection

6.4.1 In order to approve the a pplication, the Organization s hould determine that the Active Substances, Preparations or Re levant Chemicals are not Persis tent, Bioaccumulative and Toxic (PBT). Preparations that exceed all these crit eria (Persistence, Bioaccumulation and Toxicity) in

the table below are considered PBT.

The chemical analyses and toxicity assays results give no rise to concern on potential risks to the aquatic environment. As no pote ntially dangerous substances were identified in the water and the full strength effluent did not have any dele terious effects on any of the test species, the effluent would not be expected to be harmful to aquatic organism s and have negative effects on ecological communities.

Table 1 Criteria for identification of PBT substances

Criterion PBT criteria

Persistence Half-life:

> 60 days in marine water, or > 40 days in freshwater * , or > 180 days in marine sediment, or > 120 days in freshwater sediment *
Bioaccumulation BCF > 2,000 or LogPoctanol/water ≥ 3 Toxicity Chronic NOEC < 0.01 mg/l

__________________

* For the purpose of marine environmental risk assessment half-life data in freshwater and freshwater sediment can be overruled by data obtained under marine conditions.MEPC 56/2/1

ANNEX Page 15

I:\MEPC\56\2-1.doc 6.4.2 The Organization should determine the overall acceptability of the risk the Preparation may pose in its use for Ballast Water Management . It should do so by comparing the information provided and the undertaken assess ment of PBT and the discharge with scientific knowledge of the Active Substances, Preparations and Relevant Chemicals concerned. The risk evaluation should qualitatively take into acco unt cumulative effects that may occur due to the nature of shipping and port operations.

6.4.3 The risk evaluation should consider the uncertainties involved in the application for approval, and as appropriate, provide advice on how these uncertainties can be dealt with.

7 REGULATION OF THE USE OF ACTI VE SUBSTANCES AND PREPARATIONS

7.1 Handling of Active Substances and Preparations

7.1.1 The proposal for approval of Active Subs tances and Preparations should include information on their intended use and applica tion. The quantity of Active Substances and Preparations to be added to the ballast water and the maximum allowable concentration of the

Active Substances therein should be described in the instructions provided by the manufacturer.

The system is designed with a specified numb er of UV-lamps and with specified catalyst geometry. The same power will be used regardless of the water quality. The toxicity testing and chemical properties of the water are tested on th is fixed power and at sp ecified flow rate. The system can not generate more free radicals than what is achieved with the energy used.

The system should ensure that the maximu m dosage and maximum allowable discharge concentration are not exceeded at any time.

See 7.1.1.

7.2 Hazard documentation and labeling

7.2.1 The proposal should include ((M)SDS) as required. The (M)SDS should describe appropriate storage and handling to gether with the effects of degr adation and chemical reactivity during storage and should be included in the instructions provided by the manufacturer.

Not applicable.

7.2.2 Documentation of hazards or the (M )SDS should conform to the UN Globally Harmonized System of Classification and Label ling of Chemicals (GHS) and the relevant IMO regulations (e.g., the IMDG Code) and guidelin es (e.g., the GESAMP Hazard Evaluation Procedure). Where these regimes are not applicable, relevant na tional or regional regimes should

be followed.

7.3 Procedures and use

7.3.1 Detailed procedures and information for safe application of Active Substances and Preparations on board should be supplied and co mply with the approval conditions such as

maximum allowable concentration and maxi mum discharge concentration, if any.MEPC 56/2/1

ANNEX Page 16

I:\MEPC\56\2-1.doc 8 APPROVAL

8.1 Basic approval

8.1.1 All proprietary data should be treated as confidential by the Organization and its Technical Group, the Competent Authorities invol ved, and the evaluating regulatory scientists, if any.

8.1.2 Procedure to be followed:

.1 The manufacturer should evaluate the Activ e Substances or Preparations and the potential discharge in accordance with th e approval criteria specified in this procedure.

.2 Upon completion, the manufacturer should prepare an Applicat ion on the Active Substances and Preparations and submit it to the Member of the Organization

concerned.

.3 The Administration having received a sa tisfactory application should as soon as possible propose an appr oval to the Organization.

.4 Members of the Organization may propose an approval.

.5 The Organization should announce and se t the time frame for the evaluation of Active Substances and Preparations.

.6 Parties, Members of the Organizati on, the United Nations and its Specialized Agencies, intergovernmental organiza tions having agreements with the Organization and non-governmental organizatio ns in consultative status with the Organization may submit in formation that is rele vant to the evaluation.

.7 The Organization should es tablish a Technical Group in accordance with its rules of procedure ensuring that proprietary da ta should be treated as confidential.

.8 The Technical Group should review the comprehensive proposal along with any additional data submitted and report to th e Organization whether the proposal has demonstrated a potential for unreasonable risk for environment, human health, property or resources in accordance with th e criteria specified in this procedure.

.9 The Technical Group’s report should be in written form and circulated to the Parties, Members of the Organization, the United Nations a nd its Specialized Agencies, intergovernmental organiza tions having agreements with the Organization and non-governmental organizatio ns in consultative status with the Organization, prior to its consider ation by the competent Committee.

.10 The Committee of the Organization s hould decide whether to approve any proposal, introduce any modifications theret o, if appropriate, taking into account the Technical Group’s report.MEPC 56/2/1

ANNEX Page 17

I:\MEPC\56\2-1.doc .11 The Member of the Organization th at submitted the application to the Organization should inform in writing the applicant about the decision made with regard to the respective Active Substan ce or Preparation and their manner of application.

.12 Active Substances or Preparations r eceiving basic approval by the Organization may be used for prototype or type a pproval testing based on the guidelines developed by the Organization. An Active Substance or Preparation may be used for Prototype or Type Approval testi ng for the approval of different BWMS without going through ba sic approval again.

8.2 Final approval

8.2.1 In accordance with Regulation D-3.2, a Ballast Water Management system using an Active Substance or Preparation to comply with the Convention (which received basic approval) must be approved by the Organi zation. For this purpose, th e Member of the Organization submitting an application should conduct the Type Approval tests in accordance with Guidelines for Approval of Ballast Water Management System s. The results should be conveyed to the Organization for confirmation that the residual toxicity of the discharge conforms to the

evaluation undertaken for Basic Approval. This would result in Final Approval of the Ballast Water Management system in accordance w ith Regulation D-3.2. Ac tive Substances or Preparations that have receive d Basic Approval by the Organizati on may be used for evaluation of Ballast Water Management systems using Active Substances or Preparations for Final Approval.

8.3 Notification of approval

8.3.1 The Organization will record the Basic and Final Approval of Active Substances and Preparations and Ballast Water Management syst ems that make use of Active Substances and circulate the list once a year including the following information:

• Name of Ballast Water Management system that make use of Active Substances and Preparations; • Date of approval; • Name of manufacturer; and • Any other specifications, if necessary.

8.4 Modification

8.4.1 Manufacturers should report any modifications in names, including trade and technical name, composition or use of the Active Substan ces and Preparations in the Ballast Water Management systems approved by the Organization, to the Member of the Organization. The Member of the Organization should in form the Organization accordingly.

8.4.2 Manufacturers intending to sign ificantly change any part of a Ballast Water Management System that has been approved by the Organizati on or the Active Substances and Preparations used in it should submit a new application.MEPC 56/2/1

ANNEX Page 18

I:\MEPC\56\2-1.doc 8.5 Withdrawal of approval

8.5.1 The Organization may withdraw any appr oval in the following circumstances:

.1 If the Active Substances and Preparati ons or Ballast Water Management system

that make use of Active Substances no longer conforms to requirements due to

amendments of the Convention.

.2 If any data or test records differ mate rially from data relied upon at the time of approval and is deemed not to satisfy the approval condition.

.3 If a request for withdrawal of a pproval is made by the Member of the Organization on behalf of the manufacturer.

.4 If unreasonable harm to environment, human health, property or resources is demonstrated by any Member of the Organi zation or observer to have been caused by the approved Ballast Water Management system that make use of Active Substances or Preparations.

9 CONCLUSION

The PureBallast process uses UV-en ergy together with catalysts in order to produce free radicals which will react with and destroy the cell membra nes of the microorganisms causing their death. The process simulates and enhances the natural pr ocess which occurs when the sunlight hits the

surface of the seawater. There are no chemical s added and there are no chemicals formed which

can enter the ballast tanks.

The chemical testing made before and after the treatment has not i ndicated any significant

residual substances. The results from acute and chronic toxicity tests s how that there is no significant toxic effect detected in the treated water, see appendix 4.

The system placed on board will not require any ha ndling of chemicals on board, neither will it compromise the safety for the ship or the cr ew. As showed in the chemical tests the properties/reactivity of the water is not changed compared to the inlet seawater and will not

affect the corrosion rate in the BW tanks.

REFERENCES (FOR PUREBALLAST PROCESS DESCRIPTION)

The references below are all sent as documents t ogether with this application, except for the ones already sent for the Basic Approva l. The references already sent together with Basic Approval are marked in grey letters an d is available for the GESAMP group. The new references are

numbered and are filed acco rdingly in the folders.

Ahmed S., Jones C.E., Kemp T.J. and Unwin P. R. (1999) “The role of mass transfer in solution photocatalysis at a supported titanium dioxide surface” Phys. Chem. Chem. Phys., (1),

5229-5233.

Al-Ekabi H., Butters B., Dela ney D., Ireland J., Lewis N., Powwell T. and Story J. (1993) “Photocatalytic purification and treatment of water and air” E d. Ollis D.F. and Al-Ekabi H., Elsevier, Amsterdam, 321-335.MEPC 56/2/1

ANNEX Page 19

I:\MEPC\56\2-1.doc Baxendale J.H. and Busi F. (1982) “The study of fast and transi ent species by pulse radiolysis” NATO Advanced Study Institute Series , Reidel Publ. Company, Vol. D.

1. Bouchard JN, Campbell DA, Roy S (2005) “Effects of UV-B radiation on the D1 proteinrepair cycle of natural phytoplankton communities from three latitudes (Canada, Brazil, and Argentina)”. J. of Phyc. 41 (2), 273-286.

2. Buma AGJ, Helbling W, de Boer MK , Villafañe V (2001) “Patterns of DNA damage and photoinhibition in temper ate South-Atlantic picophytoplankton exposed to solar ultraviole t radiation”. J. of Photochem. and Photobiol. B: Biology 62 (1-2), 9-18.

Buxton G.V., Greenstock C.L., Helman W.P. and Ross A.B. (1988) “Critical Review of Rate

Constants for Reactions of H ydrated Electrons, Hydrogen At oms and Hydroxyl Radicals in Aqueous Solutions” J. of Phys. and Chem. Ref. Data, 2(17), 513-886.

3. Demmig-Adams B., Gilmore A.M. and Adams III W.W. (1996) “In vivo

functions of carotenoids in hi gher plants”, FASEB J 10, 403-412.

Diebold U. (2003) “The surface science of tita nium dioxide” Surface Science Reports 48,

53-229.

4. Diffey BL (1991) “Solar ultraviolet radia tion effects on biological systems”. Phys. In Med. and Biol. 36 (3): 299-328. free access via: http://www.ciesin.org/docs/001-503/001-503.html

5. Du Y. and Rabani J. (2006) “Flocculation-Induced Homolysis of Hydrogen Peroxide in Aqueous Solution of Titanium Dioxide Nanoparticles” J. Phys. Chem. B 110, 6123-6128.

Engel D.W. et al. (1974) “The effect of ionizing radiation and salinity on the grass shrimp Palaemonetes pugio” Radiation Research 59.

Fujishima A., Rao T.N. and Tryk D.A. (2000) “Titanium dioxide photoc atalysis” Journal of Photochemistry and Photobiology C: Photochemistry Reviews 1, 1-21.

6. Gao R., Safrany A., and Rabani J. (2003) “Reactions of TiO2 excess electron in nanocrystallite aqueous solutions studied by pulse and gamma-ra diolytic systems” Rad. Phys. Chem. 67, 25-39.

7. Gieskes W.W.C. and Buma A.G.J. ( 1997) “UV damage to plant life in a photobiologically dynamic environment: th e case of marine phytoplankton” Plant Ecology 128: 17-25.

8. Hashimoto K., Irie H. and Fujishima A. ( 2005) “TiO2 Photocatal ysis: A Historical Overview and Future Prospects”Japane se Journal of Applied Physics 44(12),

8269-8285.

Horne R. A. (1969) in Marine Chemistry , Wiley-Interscience, N.Y., 153-155.

9. Hessen D, De Lange H., Van Donk E (1997) “UV-induced changes in phytoplankton cells and its effects on graze rs”. Freshwater Biol. 38 (3), 513-524.MEPC 56/2/1

ANNEX Page 20

I:\MEPC\56\2-1.doc 10. Häder DP (2000) “Effects of solar UV-B radiation on aquatic ecosystems”. Adv. in Space Res. 26, 2029-2040.

11. Häder DP, Kumar HD, Smith RC, Worre st RC (2003) “Aquatic ecosystems:

effects of solar ultraviolet radiation and interactions with other climatic change

factors”. Photochem. and Photobiol. Sci. 2, 39-50.

Ivanovskii IuA. and Mitrofanov IuA (1978) “Stimu lation of Artemia salina nauplius emergence under the action of gamma irradia tion” Radiobiologiia:18(1), 135-8.

Kamat P.V. (1993) “Photochemistry on Nonre active and Reactive (Semiconductor) Surfaces” Chem. Rev. 93, 267-300.

12. Kaiser E. and Herndl G.J. (1997) “Rapid Recovery of Marine Bacterioplankton Activity after Inhibition by UV Radiation in Coastal Waters” App. And Envir. Microb. 63 (10), 4026-4031.

Legrini O., Oliveros E. and Braun A.M. (1993) “Photochemical Processes for Water Treatment” Chem. Rev. 93, 671-698.

Lindner M., Theurich J. and Bahnemann D.W. (1997) “Photocatalytic de gradation of organic compounds: accelerating the process efficiency ” Water Science and Technology, 35(4), 79-86.

13. Madsen D., Larsen J., Jensen S.K., Keiding S.R and Thøgersen J. (2003) “The primary Photodynamics of Aqueous Nitate : Formation of Peroxynitrite” Journal of the American Chemical Society 125, 15571-15576.

14. Mendez-Alvarez S., Leisinger U. and Egge n I. L. R. (1999) “Adaptive responses in Chlamydomonas reinhardtii” INTERNATL MICROBIOL, 2, 15-22.

Mills A., Belghazi A. and Rodman D. (1996) “Bromate removal from drinking water by semiconductor photocatalysis” Wat. Res. 30(9), 1973-1978.

15. Moore M.M., Breedveld M.W. and Autor A.P. (1989) “The role of carotenoids in preventing oxidative damage in the pigm ented yeast, Rhodotorula mucilaginosa”, Arch Biochem Biophys 270, 419-431.

NDRL/NIST Solution Kinetics Database (2005), Na tional Institute of Standards and Technology, webresource http://kinetic s.nist.gov/solution/index.php

Nogueira F. et al. (1998) “Radio resistance studies in Methylobacterium” Radiat. Phys. Chem. 52(1-6), 15-19.

Noguchi H., Nakajima A., Watanabe T. and Hash imoto K. (2003) “Design of a photocatalyst for bromate decomposition: Surface modification of TiO2 by pseudo-boehmite” Environ. Sci. Technol. 37, 153-157.

Oosawa Y. and Gretzel M. (1988) “Effect of surface hydroxyl density on photocatalytic oxygen generation in aqueous TiO2 suspensions” J ournal of the Chemical Society, Faraday Transactions 1, 84(1), 197-205.MEPC 56/2/1

ANNEX Page 21

I:\MEPC\56\2-1.doc Pan J., Thierry D. and Leygraf C. (1996) “Hydroge n peroxide toward enhanced oxide growth on

titaniumin PBS solution: Blue coloration and c linical relevance” J. Biomedical Materials Research, l 30, 393-402.

Platzman R. and Frank J. (1954) “The role of the hydration configuration in electronic processes involving ions in aqueous solution” The Europ ean Physical Journal A – Hadrons and Nuclei (Historical Archive), 138, 411-431.

Rijstenbil J.W. (2003) “Effects of UVB radi ation and salt stress on growth, pigments

and antioxidative defence of the marine diatom Cylindrotheca closterium” Mar. Ecol. Prog. Ser. 254, 37-48.

Rijstenbil J.W. (2005) “UV – a nd salinity-induced oxidative e ffects in the marine diatom Cylindrotheca closterium during simula ted emersion” Mari ne Biology 147:1063-1073.

Rothenberger G., Moser J., Grätzel. M., Serpone . N. and Sharma. D.K. (1985) “Charge Carrier Trapping and Recombination Dynamics in Sm all Semiconductor Particles” J. Am. Chem.

Soc. 107:8054.

16. Sarthou G., Jeandel C., Brisset L, Amour oux D., Besson T. and Donard O.F.X. (1997) “Fe and H2O2 distributi ons in the upper water colu mn in the Indian sector

of the Southern Ocean” Earth and Planetary Science Letters 147, 83-92.

17. Stenman D. (2004) “Advanced oxidation t echnologies for the pulp industry. – an investigation on the delignify ing properties of the carbonate radical.” PhD-thesis

KTH. ISBN 91-7283-903-0.

Tada H. (2002) “Photocatalytic oxidation proc esses at titanium dioxi de surfaces: enhancing

effects of surface modificati on with oxide-(sub) monolayer s on photocatalytic activity” Encyclopedia of surface and colloid sc ience, Marcel Dekker Inc., 4043-4058.

Treinin A. and Hayon E. (1975) “Charge Transf er Spectra of Halogen Atoms in Water.

Correlation of the Electronic Tran sition Energies of Iodine, Bromine, Chlorine, Hydroxyl, and Hydrogen Radicals with Their Electron Affi nities” J. Am. Chem. Soc. 97(7):1716-1721.

Xiao-Ying Yu (2004) “Critical Eval uation of Rate Constants of H ydrogen Peroxide Photolysis in Acidic Aqueous Solutions Containing Chlori de Ions” J. Phys. Ch em. Ref. Data, 33(3).

Zaafrane S., Maatouk K., Gauthier J.M. and Bakhrouf A. (2004) “Influence of preliminary cultivation conditions and the presence of th e rpoS gene on the su rvival of Salmonella typhimurium in sea water exposed to sunli ght” Canadian Journal of Microbiology 50(5),

341-350.

18. Woodward J. R., Cirillo V. P. and Edmunds L. N. JR. (1978) “Light Effects in Yeast: Inhibition by Visible Light of Gr owth and Transport in Saccharomyces cerevisiae Grown at Low Temperatures” J. of Bacteriology, 133(2), 692-698.MEPC 56/2/1

ANNEX Page 22

I:\MEPC\56\2-1.doc APPENDIX 1 – LLOYDS APPROVAL OF FULL-SCALE PROTOTYPE SYSTEM ONBOARD M/V DON QUIJOTE

APPENDIX 2 – EXTRACTION FROM QAPP FOR FULL SCALE TEST PROGRAM

Due to the bulkiness only the table of contents and short description of the QAPP is included. The QAPP is has been approved by DNV prior to the start of the Type Approval tests.Norwegian Institute for Water Research

Revised: 6 th of December 2006 Approved: 6 h of December 2006



# Extracted version

Quality Assurance Project Plan (QAPP)

- for the full scale testing of th e PureBallast treatment system of

Alfa Laval/Wallenius Water ABTable of contents

Abbreviations and acronyms

1. PROJECT DESCRIPTION, OBJECTIVES AND ORGANISATION

1.1 Address of QAPP

1.2 Processes to be evaluated 1.2.1 Preparations

1.2.2 Execution of tests 1.2.3 Reporting of results

1.3 Description of test site and technology set-up
1.3.1 Test site 1.3.2 The PureBallast water management system (PureBallast system) 1.3.2.1 Process description 1.3.2.2 System and component description 1.3.2.3 Physical properties of the PureBallast test equipment

1.4 Organization of the project 1.4.1 Presentation of Norwegian Institute for Water Research (NIVA) - testing site owner and testing organization 1.4.2 Quality management plan (QMP) of NIVA
1.4.3 Presentation of the vendor

1.4.3.1 Presentation of Alfa Laval

1.4.3.2 Presentation of Wallenius Water AB

1.5 Responsibilities of a ll project participants 1.5.1 Preparation of test tanks and clar ification for the installation of the PureBallast treatment system at the test facility 1.5.2 Installation, clarification, operation and dismantling of the PureBallast treatment system 1.5.3 Fulfilment of the chemical water quality requirement 1.5.4 Fulfilment of the biological water quality requirement 1.5.5 Sample collection and preservation 1.5.6 Clarifying test waters for discharge to recipient waters 1.5.7 Laboratory chem ical measurements 1.5.8 Measurements of biological treatment factors 1.5.9 Acute and chronic toxicity tests 1.5.10 Data handling and reporting of test results 1.5.11 Project management and coordination 1.5.12 Quality assurance of project

2. EXPERIMENTAL APPROACH

2.1 Technology installation and shakedown procedures

2.2 Technology start-up and stop procedures 2.2.1 System start-up procedures 2.2.2 System stop procedures

2.3 Technology ca libration checks

2.4 Preparation of test waters 2.4.1 Assurance of fulfilment of chemical water quality test criteria 2.4.2 Assurance of fulfilment of biological water quality test criteria 2.4.2.1 Harvesting of indigenous organisms
2.4.2.2 Cultivation of Artemia salina and Tetr aselmis suecia and/or Brachiomonas submarina

2.4.2.3 Bacteria2.4.3 Discharge of waters

2.5 Running test cycles

2.6 Sampling and measurements

2.7 Time schedule for the testing period

3. SAMPLING PROCEDURES

3.1 Representativeness of samples

3.2 Sampling of test waters

3.3 Sample preservation

3.4 Measures to avoid cross-contamination during test water transfer and sampling

4. TESTING AND MEASUREMENT PROTOCOLS

4.1 In situ measurements 4.1.1 Temperature 4.1.2 pH 4.1.3 Dissolved oxygen (DO) 4.1.4 Salinity

4.2 Discrete samples 4.2.1 Dissolved organic carbon (DOC) 4.2.2 Particulate organic carbon (POC) 4.2.3 Total suspended solids (TSS) 4.2.4 Organisms >50 μ m 4.2.5 Organisms ≥ 10-50 μ m 4.2.6 Heterotrophic bacteria

4.2.7 Coliforms

4.2.8 E.coli 4.2.9 Enterococcus group
4.2.10 Intestinal Enterococci

4.2.11 Vibrio cholerae

4.2.12 Vibrio cholerae (Serotypes O1 and O139)

4.3 Chemical measurements – initial quick tests 4.3.1 Dissolved organic carbon (DOC) 4.3.2 Particulate organic carbon (POC) 4.3.3 Total suspended solids (TSS) 4.3.4 Turbidity

4.4 Toxicity testing of treated ballast water 4.4.1 Growth inhibition of marine algae 4.4.2 Acute toxicity of marine copepods 4.4.3 Reproduction of marine copepods 4.4.4 Acute toxicity of fish 4.4.5 Juvenile fish growth test

5. QA/QC CHECKS

5.1 Data Quality Indicators (DQIs) 5.1.1 Representativeness 5.1.2 Accuracy
5.1.3 Precision5.1.4 Bias 5.1.5 Comparability

5.2 Emergency plan
5.2.1 Power failure 5.2.2 Collapse in organism cultivation 5.2.3 Pumping failure

5.2.4 Identification of Vibrio cholerae in test water 5.2.5 The right to veto between verifier and project group

6. DATA REPORTING, DATA REDU CTION AND DATA VALIDATION

6.1 Approach for data ma nagement and evaluation

6.2 Reporting requirements

6.3 References

Appendix A – Total project management

Appendix B – Chemical water quality preparation

Appendix C – Biological water quality preparation

Appendix D – Collection and preservation of samples

Appendix E – Logging of in situ measurements

Appendix F – Evaluation form for organisms >50 μm

Appendix G – Evaluation form for organisms ≥ 10-50 μm

Appendix H – Evaluation form for heterotrophic bact eria, coliform bacteria, En terococcus group bacteria and Vibrio sp.

Appendix I – Acute toxicity of the green al gae Skeletonema costatum (page 1 of 1)

Appendix J – Acute toxicity of ma rine copepods (page 1 of 1)

Appendix K – Acute toxicity of juven ile turbot; setup form for method K19

Appendix L – Process Description

Appendix M – Description and the PureB allast system and its components1. Project description, ob jectives and organisation 1.1 Address of QAPP This QAPP describes the implementation of quality assurance and quality control activities during the

evaluation of the Alfa Laval Pureballast water ma nagement system according to the requirements for biological and toxicity testing stated in the IMO Guidelines for Approval of Shipboard Ballast Water Treatment Systems (MEPC 53/24/Add.1, Annex 3, 2005). The requirements involve all parts and processes of the test as listed in: Annex - Guidelines for approval of balla st water management systems (G8)

• Part 2 – Test and performance specifications for approval of ballast water management
systems

• 2.1 Quality assurance and quality control procedures (page 15)

• 2.3 Land-based Testing (pages 17-22)

• 2.4 Reporting of Test Results (page 22)

• Part 4 – Sample analysis methods for the dete rmination of biological constituents in ballast

water

• Section 4.1 – 4.8 (page 24-26)

Annex 4 MEPC 53/25/Add.1 - Procedure for approval of ballast water management systems that make use of active substance (G9)

• Section 1-5 (pages 1-7)

The QAPP is a mean to reveal any problems before st art-up and during execution of the project at as early stage as possible to minimize any potential procedur al, technical and scientific inadequacies and timeand economic losses. This QAPP will be used for biological testing in accordance with the IMO guidelines as well as for the final and basic approval testing of active substances.APPENDIX 3 – Assessment report Det Norske VeritasAPPENDIX 4 – Full scale toxicity testing of the PureBallast system



## REPORT SNO



# F ull scale toxicity testing of the PureBallast System

- preliminary reportNIVA



## Norwegian Institute for Water Research

– an institute in the Environmen tal Research Alliance of Norway 

# REPORT

Main Office Regional Office,
Sørlandet Regional Office, Østlandet Regional Office,
Vestlandet Regional Office Midt-Norge

P.O. Box 173, Kjelsås Televeien 3 Sandv ikaveien 41 Nordnesboder 5 P.O. Box 1266 N-0411 Oslo, Norway N-4879 Grimstad, Norway N-2312 Ottestad, Norway N-5008 Bergen, Norway N-7462, Norway Phone (47) 22 18 51 00 Phone (47) 37 29 50 55 Phone (47) 62 57 64 00 Phone (47) 55 30 22 50 Phone (47) 73 54 63 85 / 86 Telefax (47) 22 18 52 00 Telefax (47) 37 04 45 13 Telefax (47) 62 57 66 53 Telefax (47) 55 30 22 51 Telefax (47) 73 54 63 87
Internet: www.niva.no

Title Full scale toxicity testing of the PureBallast System – preliminary report Serial No.

Report No. Sub-No. Date
2006

Pages Price

Author(s) Helge Liltved Christian Vogelsang August Tobiessen
Tor Gunnar Jantsch Topic group

Water treatment

Geographical area Norway Distribution
Confidential

Printed
NIVA

Client(s) Wallenius Water AB, Stockholm, Sweden Client ref.
Carl Tullstedt

Abstract The PureBallast System, developed by Alfa Laval / Wallenius Water, was tested in a full scale system to evaluate the performance according to the IM O Guidelines. The biological and toxicologi cal tests were conducted at NIVA’s test site located at Solbergstrand 20 km south of Oslo. Test cy cles were conducted with moderate and high salinity water. Each cycle lasted for a period of 5 days. Based on the conducted acute and chronic to xicity tests, and chemical analyses, it is concluded that ballast water treated with the present technology do not provide hazard or risk to the marine ecosystem when released.

4 keywords, Norwegian 4 keywords, English

1. Ballastvann 1. Ballast water

2. Toksikologi 2. Toxicology

3. Testing 3. Testing

4. Wallenius Water AB 4. Wallenius Water AB

Helge Liltved HelgeLiltved Jarle Nygard
ISBNNIVA

4 Contents

Abbreviations and acronyms Summary

1. BACKGROUND

2. MATERIALS AND TEST PROTOCOLS

2.1 Test site

2.2 The evaluated ballast water treatment system 2.2.1 General description of the PureBallast System

2.3 Test waters 2.3.1 Overview of requirements 2.3.2 Fulfilment of chemical water quality test criteria 2.3.3 Fulfilment of biological water quality test criteria Harvesting of indigenous organisms Cultivation of Artemia salina and Brachiomonas submarina

Bacteria

2.4 Test procedure 2.4.1 Description of test cycle 2.4.2 Measures to avoid cross-contamination during water transfer and sampling

2.5 Sampling 2.5.1 Assuring the representativeness of samples 2.5.2 Sampling protocols 2.5.3 Overview of sampling equipment 2.5.4 Sample preservation

2.6 Analyses
2.6.1 Chemical measurements Temperature pH Dissolved oxygen (DO) Salinity Dissolved organic carbon (DOC) Particulate organic carbon (POC) Total suspended solids (TSS) 2.6.2 Identification and quantification of viable organisms ≥ 50 μm 2.6.3 Identification and quantification of viable organisms >10-50 μm 2.6.4 Identification and quantification of viable bacteria Heterotrophic bacteria Thermotolerant coliform bacteria Enterococcus group Vibrio sp. 2.6.5 Toxicity measurements of treated Ballast water

3. RESULTS AND DISCUSSION

3.1 Fulfillment of chemica l water quality criteriaNIVA

5 3.2 Fulfillment of the biological water quality criteria

3.3 Operational performance of the PureBallast system

3.4 Biocidal effects on organisms ≥ 50 μm in minimum diameter

3.5 Biocidal effects on organisms ≥ 10-50 μm in minimum diameter

3.6 Bactericidal effects

3.7 Ecotoxicological responses Appendix A. Test reportsNIVA

6 

### Summary

The tests were conducted at NIVA’s test site lo cated at Solbergstrand 20 km south of Oslo. The tests were conducted with moderate salinity and with high salinity. Each cycle lasted for a period of 5 days. The tests were conduc ted according to the IMO specifications.

The test water was prepared in a common 516 m 3 tank (WST). A combination of indigenous harvested organisms and cultured surrogate species (>50 μm group: Artemia salina ; 10-50 μm

group: Brachiomonas submarina and/or Tetraselmis suecica ) and heterotrophic bacteria were added to fulfil the biolog ical water quality criteri a described in Table 1.

The test line involved consecutive treatment of at least 200 m 3 test water by the 50 μm filter unit and the PureBallast System transferring th e test water from WST to a temporary holding tank (TT1) before a second treatment by the PureBa llast System and transfer to a storage tank (TT2) for at least five days storage. The c ontrol line was run by tran sferring at least 200 m 3 of the same type of prepared test water from WST to a second temporary holding tank (TT1) using the pump of the PureBallast system, but in by-pass of the treatment units before transfer to a second storage tank (CT2) fo r a five days storage time.

Fulfilment of the chemical and biol ogical requirements of the test water

• The required levels for total suspended so lids (TSS), dissolved organic carbon (DOC) and particulate organic carbon (POC) were fulfilled in all test cycles reported.

• The requirements regarding density of the ≥ 50 μm group was met in the tests reported. The requirements regarding the biological di versity within the population were most probably fulfilled in all tests. The final analysis has not been conducted yet.

• The requirement for the ≥ 10-50 μm group was met. The requirements regarding the biological diversity within the population were most probably fulfilled in all tests. The final analysis has not been conducted yet.

• The only requirement regarding bacteria in infl uent water is that he terotrophic bacteria should be present in a density of ≥ 10 4 ml -1 . This requirement was fulfilled in all tests

Biocidal effects of treatment and storage

• The required less than 10 viable organisms ≥ 50 μm in minimum diameter per m 3 in the treated water after five days storage wa s fulfilled in all tests. The equivalent requirement of the non-treated control test water, stating that the level of viable organisms after 5 days storage should be higher than 100 per m 3 , was also fulfilled in

all tests.

• The required less than 10 viable organisms ≥ 10 - 50 μm in minimum diameter per ml

in the treated water after five days storage was reached in a ll tests, both analyzed with the dilution method and by microscope count s. The equivalent requirement for the

non-treated control test water after 5 days storage states that the level of viable organisms should be higher than 100 per ml. This requirement was fulfilled in all test by both methods.

• Regulation D-2 only requires documenta tion of maximum allowable effluent concentration after five days storage of Escherichia coli

, Vibrio cholerae (toxicogenic serotypes O1 and O139) and Enterococci ; <250 cfu/100ml, <1 cfu/100ml and <100 cfu/100ml, respectively. These requirements were fulfilled for E. coli and Enterococci

.NIVA

7 Testing to confirm any presence of serotypes O1 and O139 among the residual Vibrio sp. in treated test waters after five da ys storage is conducte d. Presence of these serotypes have not been reporte d in Norwegian coastal water.

Toxicity testing The ecotoxicity of treated ballast water wa s tested once with undiluted (100 %) samples on three trophic levels using marine chronic tests with untreated control water as control. No detectable toxicity was observed on the marine alga Skeletonema costatum (growth inhibition test), Nitocera spinipes (invertebrate reproduction test) or Scophtalmus mixmus (acute fish test and fish growth test). A mortality of 27% of the marine crustacean Acartia tonsa was observed at the highest tested concentration (100 %) compared to 9% in the control. It should be commented that high death-rate in the cont rol can be a problem with the test, as also experienced in the current test. This death-rate can give misleading results. No effects were

observed in the test with th e other marine crustacean Nitocra spinipes . It is therefore doubtful

that the effect observed can be attributed to toxicity. Lack of mortality >50 % at any test concentration precludes estimation of LC 50

.

Treated waters in test cy cle 3 were also sampled and analysed for AOX, EOX, trihalomethanes, Potential Bioaccumulating Substances (PBS) and free and total residual oxidants (TRO). All compounds, except AOX and PBS, were below the detection limits. However, the concentrations of AOX and PB S were not significant different from the

concentrations found in control water.

Based on the conducted acute and ch ronic toxicity tests, and th e chemical analysis, it is

concluded that ballast water treated with the present ballast water t echnology do not provide hazard or risk to the marine ecosystem when released.NIVA

8 1 Background

The overall goal for Alfa Laval /Wallenius Water is certification of the PureBallast Ballast Water Management System in accordance with the requirements in the IMO Convention on ballast water management and underlying guide lines, including the IMO Guidelines for Approval of Ballast Water Management Sy stems, Annex 3 and Annex 4 (july 2005).

Pilot tests were conducted, in J une 2006, for partial fulfilment of such a certification. The objective of the pilot testing was to verify th e environmental performance characteristics of the PureBallast Pilot System technology rela tive to the biological and toxicological requirements stated in the IMO Guideline. By performing a pilottest, difficulties and “bottlenecks” could be detected, and measures could be taken to avoid probl ems during the full scale testing. The experience from the pilottest served as a fundament for te sting of the technology

in full scale.

These full scale Type Approval tests were made in accordance with the requirements in the IMO Convention on ballast water management a nd underlying guidelines, including the IMO Guidelines for Approval of Ballast Water Mana gement Systems, Annex 3 and Annex 4 (july 2005).

2 Materials and test protocols

2.1 Test site

The tests were conducted at NIVA’s test site (s ee Figure 1) located at Solbergstrand 20 km south of Oslo. Sea water is supplied from various depths down to 60 m in the Oslofjord, while fresh water is supplied from ground water bore holes or from a local creek.

The test facility includes 4 glass-fibre reinfo rced polyester tanks, s upplied with inlet and outlet arrangements and equipment for proper cleaning. Two of the tanks are covered to prevent light introduction. The surfaces of the test tanks (TT1 and TT2) and the control tank (CT1) are coated with co atings for ships (Balloxy HB light , Jotun, Norway). Propeller devices with gentle rotation are mounted at the bottom and at shallow depth in WST and TT1, and at the bottom in TT2 and CT2. These propeller devices are used to suspend particles, including algae and zooplankton, evenly and homogenise the content. A te st is conducted to verify homogenisation prior to each test and prior to sampling.

2.2 The evaluated ballast water treatment system

2.2.1 General description of the PureBallast System
The PureBallast System from Alfa Laval / Wallenius Water is a complete ballast water treatment system composing of three main comp onent ballast water treatment system; 1) a 50 μm filter for removal of larger particles and or ganisms, 2) the Wallenius AOT unit, a patented treatment technology using th e synergetic effects of in situ produced free radicals and directNIVA

9 photo-radiation to inactivate microbes and 3) control and auxiliar y equipment including sampling points to control water flows and meas ure different alarm levels during operation as well as to take samples during operation and test ing. A detailed description of the PureBallast system is given in the QAPP.

2.3 Test waters

Test waters were prepared in a 516 m 3 tank (WST) from high salinity sea water from 60 meters depth or from brackish surface water depending on the required salinity (>32 PSU or 3-32 PSU, respectively, with a minimu m difference of 10 PSU). The 516 m 3 of test water was used for both testing and control. A combin ation of harvested i ndigenous organisms and cultured surrogate species (>50 μm: Artemia salina ; 10-50 μm: Tetraselmis suecica and/or

Brachiomonas submarina and heterotrophic bacteria) were added to fulfil the biological water quality criteria, and soluble lignin, starch and kaolin was adde d to adjust the contents of dissolved organic carbon (DOC), particulate organic carbon (POC) and total suspended solids (TSS), respectively, to within the limits of chemical water quality criteria.

2.3.1 Overview of requirements
Table 1 summarizes the chemical requirements of the test waters, while Table 2 summarizes the biological requirements.



### Table 1. Required chemical water quality of test waters . Salinities should be separated by at least 10

PSU Salinity DOC POC TSS

Test water 1 >32 PSU >1 mg/l >1 mg/l >1 mg/l Test water 2 3-32 PSU >5 mg/l >5 mg/l >50 mg/l

Test water 3 ≤ 3 PSU >5 mg/l >5 mg/l >50 mg/l



### Table 2. Required biological water quality in influent t est water, treated water and in control after 5 days storage as stated in regulation G8 by IMO.

Organism group Influent water In treated water after 5 days storage In control after 5 days storage**

≥ 50 μm min.

dimension Pref. 10 6 m -3

, ≥ 10 5 m -3
Min. 5 species from 3

diff. phyla/divisions <10 viable organisms per m 3 > 10x <10 viable organisms per m 3

≥ 10-50 μm min.

dimension 10 4 ml -1

, ≥ 10 3 ml -1
Min. 5 species from 3

diff. phyla/divisions <10 viable organisms per m 3 > 10x <10 viable organisms per m 3

Heterotrophic

bacteria ≥ 10 4 ml -1

Vibrio sp. <1 cfu/100 ml ≥ 10x <1 cfu/100 ml

Escherichia coli <250 cfu/100 ml ≥ 10x <250 cfu/100

ml

Intestinal

Enterococci <100 cfu/100 ml ≥ 10x <100 cfu/100

ml

** As discussed in the results and discussion section.NIVA

10 2.3.2 Fulfilment of chemical water quality test criteria
Soluble lignin (40 % solution, Borrebond FP-P, Bo rregaard, Norway), starch and kaolin clay (Merck) was added to adjust the contents of dissolved organic carbon (DOC), particulate organic carbon (POC) and total suspended solids (TSS), respectively, to within the limits of chemical water quality criteria described in Table 1. The necessary additions of each compound were calculated on the ba sis of earlier laboratory tests where the actual addition to each parameter (e.g. DOC, POC and TSS) one compound unit resulted in were measured. Initially it was intended to use rapid test kits to measure both DOC, POC and TSS right addition, however, the mandatory long heating tim e (2 hours) in sulphuric acid to break down organic compounds for the DOC and POC measur ements disqualified this option due to logistic problems. Hence, only the TSS criteri a were checked by direct measurement on site, using the HACH method 8006 (HACH, 2001), app licable to samples with a TSS content within the range 0-750 mg/l, and the pre-set calibration st andards of the Odyssey DR2500 spectrophotometer (HACH).

2.3.3 Fulfilment of biological water quality test criteria
A combination of indigenous harvested organi sms and cultured surrogate species (>50 μm:

Artemia salina ; 10-50 μm: Brachiomonas submarina and/or Tetraselmis suecica ) and heterotrophic bacteria was added to fulfil the biological water quality criteria described in

Table 2.

2.4 Test procedure

2.4.1 Description of test cycle
The water transfer pathways between tanks and the PureBallast System during each test cycle (including the control) is shown in Figure 1. The test line (blue lines) involved consecutive

treatment of at least 200 m 3 test water by the 50 μm filter unit and the Wallenius AOT unit transferring the test water from tank with prep ared test water (WST) to a temporary holding tank (TT1) before a second treatment by the Wa llenius AOT unit and transfer to a storage tank (TT2) for at least five days storage. The control line (red lines) wa s run by transferring at

least 200 m 3 of the same type of prepared test wa ter from WST to the temporary holding tank (TT1) using the pump of the PureBallast system, but in by-pass of the treatment units before transfer to a second storage tank (CT2 ) for a five days storage time.NIVA

11 Figure 1. Transfer of test water during one test cy cle with the PureBallast system including a test line (blue) and a control line (red). Sampling numbers are indicated by S1-S5.

The test cycles were conducted with high salinity water. Each cycle lasted for a period of 5 days.

2.4.2 Measures to avoid cross-contamination during water transfer and sampling

To avoid cross-contamination between consecuti ve test waters upon transfer between tanks, all pipelines were flushed for 2- 3 min with drainage to sea at normal operating flow rate, prior to transfer of water to holding or st orage tanks. Since the same holding tank

To avoid cross-contamination during sampli ng the propeller, buckets, siphon overflow and plankton net were rinsed in sea wate r from 60 meters between each sampling.

2.5 Sampling

2.5.1 Assuring the representativeness of samples
Before any samples were collected from any of the tanks at every sampling time the following procedure was carried out to assure that representative samples were withdrawn: 1) The particulate content was homogenized us ing a hand-held outboa rd propeller of 3 hp or x hp at moderate speed directed alternat ely between the periphery and centre of the

tank. 2) The turbidity in different sections of each tank (upper part, middle part and bottom part, in both periphery and mid section) was measured by a handheld submersible probe (YSI – 600 OMS) during homogenizati on. When any turbidity measurement was within a 10 % deviation from the averag e turbidity of all measurements in the tank, sampling was started. WST

516 m 3 TT1

231 m 3 TT2

205 m 3

CT2

205 m 3 S1 S2, S3

S4, S5 S4*, S5* 50 μm filter Pump
Benrad AOT ModuleNIVA

12 2.5.2 Sampling protocols

2.5.3 Overview of sampling equipment
An overview of sampling equipment and containe rs used and sampled volumes are given in

Table 3.



### Table 3. Equipment and containers used for sampling and necessary and sampled volume for the individual parameters.

Parameter Sampling equipment Sample container Sample

volume WST, CT2 Sample volume

TT2

Temperature Temp. meter - - - pH pH probe - - - Dissolved oxygen DO probe - - - Salinity Probe - - - DOC Directly, grab Acid washed glass

bottle 100 ml 100 ml

POC Directly, grab Acid washed glass

bottle 100 ml 100 ml

TSS Directly, grab Clean plastic bottle 1000 ml 1000 ml

Organisms ≥ 50 μm Sieving* 20 l 1 m 3

Organisms ≥ 10-50

μm Directly, grab Clean glass bottle 1000 ml 1000 ml

Heterotrophic

bacteria

Coliform bacteria Enterococcus group
bacteria

Vibrio sp. Vibrio cholerae Directly, grab Sterile glass bottle 1000 ml 1000 ml

* A 20-litre grab sample (f rom WST and CT2) or 1 m 3 collected through a siphon spillway (from TT2) is concentrated to a volume of 40- 100 ml through a plankt on net with diagonal dimensions of 50 μm.NIVA

13 2.5.4 Sample preservation
Preservation methods and expect ed storage/holding times before measurement are shown in Table 4. All preservation and analysis were in accordance with the described methodology.

Table 4. Preservation methods and expected st orage/holding times before measurement (ISO/CD 5667-3, 2001).

Parameter Preservation Max holding

time Expected storage

time Temperature - - 0 pH - - 0 Dissolved oxygen - - 0 Salinity - - 0 Dissolved organic carbon (DOC) Acidify to pH<2 with 1 ml 4 M H 2 SO 4 per 100 ml, stored at 4 o C 7 days 0-5 days

Particulate organic carbon (POC) Acidify to pH<2 with with 1 ml 4 M

H 2 SO 4 per 100 ml,stored at 4 o C in

darkness 7 days 0-5 days

Total suspended solids (TSS) Stored at 4 o C in darkness 24 hours <24 hours

Organisms ≥ 50 μm Stored at 4 o C in darkness 6 hours < 2 h Organisms ≥ 10-50 μm Stored at 4 o C in darkness 6 hours < 6 h Heterotrophic bacteria

Thermotolerant

coliform bacteria Enterococcus group
bacteria

Vibrio sp. Vibrio cholerae 4 o C in darkness 6 hours < 6 h

2.6 Analyses

2.6.1 Chemical measurements

Temperature Temperature was measured in situ using a calibrated thermometer.

pH pH was measured in situ using a calibrated probe and pH-meter.

Dissolved oxygen (DO) Dissolved oxygen (DO) was measured in situ using a calibrated probe and meter. DO is reported as mg O 2 /l.

Salinity Salinity was measured in situ using a calibrated salioterm.NIVA

14 Dissolved organic carbon (DOC)
DOC was measured by an accredited method based on Norwegian Standard NS-ISO 8245 (NIVA method G5-1) at NIVA after filtering the sample th rough a GF/F filter (0.7 μm). The sample was acidified with phosphoric acid and aerated with ox ygen to remove inorganic carbon prior to injection into a quartz tube filled with a platinum catalyser at 680 o C. The organic carbon compounds were

oxidized to CO 2 which was quantified using an NDIR detect or (Phoenix 8000 TOC-TC analyser with sample carousel STS 8000) with oxygen as carri er. Detection limit was 0.5 mg C/l.

Particulate organic carbon (POC)

POC is calculated as the difference between the level of total organic carbon in the sample, measured on the non-filtered sample (see 4.2.1), and the measured DOC level of the same sample.

Total suspended solids (TSS) A nucleopore capillary filter (0,4 μm) was dried at 40-50 o C for 2 hours and the tara wes determined by weighing on a micro weight (S artorius 4503 Micro) equipped with an ion source (Static Eliminator Bar Pu 210 Item no. LC 9793) removing sta tic electricity. The sample was filtered by suction through the filter, which again was dried at 40-50 o C for 2 hours before being weighed on the micro wei ght. The TSS is represented by the weight increase. Lowest reported value: 0.1 mg/l.

Trihalomethanes (THM) THMs were extracted in pentane and measured by the use of a gas chromatograph (American Public Health Association, 1989).

2.6.2 Identification and quantification of viable organisms ≥ 50 μm

2.6.3 Identification and quantification of viable organisms >10-50 μm

2.6.4 Identification and quantification of viable bacteria

2.6.5 Toxicity measurements of treated Ballast water
The following standard tests were perf ormed on the treated ballast water. -ISO 10253: Marine algal gr owth inhibition test - Forslag til Dansk Standard: Økotoksi kologisk undersøgelse med krepsdyret Nitocra spinipes . Reproduksjonsmetode -OECD Guidelines for testing of chemical s (No. 203; Fish, acute toxicity test). -OECD Guidelines for testing of chemicals" (No. 215; Fish, juvenile growth test), adapted for marine species

The methods used are given in the test re ports which are include d in the appendix 1.NIVA

15 3 Results and discussion

3.1 Fulfillment of chemical water quality criteria

The temperature, salinity, pH a nd dissolved oxygen content were measured in the test water tank (WST) prior to treatment and daily in the storage tanks TT2 and CT2 in each test cycle. Measurements were not conducted during week -ends. The results are given in Table 5.

As shown, the temperature in the initial water (WST) va ried among the tests. Smal decreses in oxygen concentration were record ed during the test cycles. Ho wever, the oxygen content did never reach critical levels regardi ng survival of test organisms.

Table 5. Temperature, salinity and dissolved oxyge n measurements in the test water tank (WST) prior to treatment and daily in the st orage tanks TT2 and CT2 for each test cycle (measurements were not conducted during week-ends).

WST TT2 CT2 Day 0 0 1 2 3 4 5 0 1 2 3 4 5

Parameter Unit Test cycle 1 Temperature oC 9.8 9.8 8.80 8.30 7.40 7.20 6.90 - 7.90 7.50 6.70 6.50 6.30 Salinity PSU 32.2 32.2 32.0 31.8 31.8 31.9 31.9 - 32.0 31.9 31.9 31.9 32.0 pH 7.8 7.8 7.50 7.43 7.49 7.44 7.40 - 7.48 7.46 7.41 7.47 7.42

Dissolved

O 2 % 85.0 85.0 83.10 78.10 78.60 73.60 72.80 - 84.30 73.20 76.50 71.10 70.40

Test cycle 2 Temperature oC 7.60 7.60 7.00 - - 5.70 5.40 - 6.70 - - 5.30 5.00 Salinity PSU 32.60 32.60 32.60 - - 32 .40 32.30 - 32.80 - - 32.60 32.30 pH 7.90 7.90 7.87 - - 6.80 7.88 - 8.01 - - 6.02 7.84

Dissolved

O 2 mg/l 10.60 10.60 7.80 - - 7.80 6.28 - 6.68 - - 7.30 7.80

Test cycle 3 Temperature oC 12.30 12.50 12.80 11. 60 - - 10.60 13.00 11.50 11.10 - - 9.80 Salinity PSU 32.60 32.60 32.60 32.60 - - 32.60 32.60 32.60 32.60 - - 32.60 pH 8.03 8.04 8.01 7.91 - - 7.82 8.03 7.98 7.93 - - 7.87

Dissolved

O 2 mg/l 9.90 11.00 8.90 8.70 - - 6.90 9.90 8.90 8.70 - - 6.80

Test cycle 4 Temperature oC 12.00 11.90 11.80 11. 70 - - 10.10 11.40 11.50 11.10 - - 9.80 Salinity PSU 32.70 32.70 32.70 32.80 - - 32.70 32.70 32.70 32.70 - - 32.70 pH 8.06 8.06 8.05 7.96 - - 7.75 8.08 8.08 7.96 - - 7.79

Dissolved

O 2 mg/l 10.30 9.40 9.30 8.50 - - 5.10 9.60 9.10 8.50 - - 6.20

Total suspended solids (TSS), dissolved organi c carbon (DOC) and partic ulate organic carbon (POC) measurements at day 0 in WST, TT2 and CT2 and at day 5 in TT2 and CT2 are shown in Table 6 (test cycle 1), Table 7 (test cycle 2) , Table 8 (test cycle 3) and Table 9 (test cycle 4). The required level of TSS, DOC and POC in the initial test water each test (as defined in Table 1) are given on the line above the measured values for the influent water to treatmentNIVA

16 (WST). The required levels were fulfilled in all test cycles as indicated by the green coloured background.



### Table 6. Test 1. Chemical water quality (average a nd standard deviation of triplicate samples). Green background indicates that required level was fulfilled, yellow background partial fulfilment, while red background indicates failure to fulfil re quired level. Compounds were added to ensure fulfilment of the requirements; both a POC regulating compound (POC Additive ) and a DOC regulating compound (DOC Additive ) was added. TSS mg/l DOC, mg/l POC, mg/l TEST CYCLE 1 Average Stdev Average Stdev Average Stdev

Additions to influent water - DOC Additive = 1.2 kg. POC Additive = 15.0 kg Required level >1 - >1 - >1 - Influent water (WST) 51.3 3.9 2.6 0.2 4.8 2.8



### Table 7. Test 2. Chemical water quality (average a nd standard deviation of triplicate samples). Green background indicates that required level was fulfilled, yellow background partial fulfilment, while red background indicates failure to fulfil re quired level. Compounds were added to ensure fulfilment of the requirements; both a POC regulating compound (POC Additive ) and a DOC regulating compound (DOC Additive ) were added. TSS mg/l DOC, mg/l POC, mg/l TEST CYCLE 2 Average Stdev Average Stdev Average Stdev

Additions to influent water - DOC Additive = 1.2kg POC Additive = 7.2 kg Required level >1 - >1 - >1 - Influent water (WST) 6.6 2.2 2.4 0.1 1.5 1.1



### Table 8. Test 3. Chemical water quality (average a nd standard deviation of triplicate samples). Green background indicates that required level was fulfilled, yellow background partial fulfilment, while red background indicates failure to fulfil re quired level. Compounds were added to ensure fulfilment of the requirements; both a POC regulating compound (POC Additive ) and a DOC regulating compound (DOC Additive ) were added. TSS mg/l DOC, mg/l POC, mg/l TEST CYCLE 3 Average Stdev Average Stdev Average Stdev

Additions to influent water - DOC Additive = 1.2kg POC Additive = 4.8 kg Required level >1 - >1 - >1 - Influent water (WST) 16.5 0.9 2.0 0.1 2.9 0.1



### Table 9. Test 4. Chemical water quality (average a nd standard deviation of triplicate samples). Green background indicates that required level was fulfilled, yellow background partial fulfilment, while red background indicates failure to fulfil re quired level. Compounds were added to ensure fulfilment of the requirements; both a POC regulating compound (POC Additive ) and a DOC regulating compound (DOC Additive ) were added. TSS mg/l DOC, mg/l POC, mg/l TEST CYCLE 4 Average Stdev Average Stdev Average Stdev

Additions to influent water - DOC Additive = 1.2kg POC Additive = 7.2 kg Required level >1 - >1 - >1 - Influent water (WST) 18.1 1.3 3.7 0.9 3.0 1.8NIVA

17 Treated waters in test cy cle 3 were also sampled and analysed for AOX, EOX and trihalomethanes, a group of halogenate d organic compounds. These compounds were analysed to clarify if the technology was cap able of oxidizing organic substances to trihalomethanes. The dominating THM found in oxi dized seawater is br omoform, which is a probable human carcinogen (USEPA 1996), known to bio-accumulate in aquatic animals. Bromoform is highly persiste nt in the environment.

No THMs were found in the treated water. All the congeners were belo w the detection limit, in both test waters sampeled (Table 9).



### Table 9. Concentration of AOX, EOX, chlorophenols a nd trihalomethanes in treated water from Test 3 in control water and treated water immediately after treatment. Unit Control water Treated water Inaccuracy % AOX mg/l 0.45 0.57 20 EOX mg/l <0.050 <0.050 20 2-Monochlorophenol μg/l <0.10 <0.10 3-Monochlorophenol μg/l <0.10 <0.10 4-Monochlorophenol μg/l <0.10 <0.10 2,3-Dichorophenol μg/l <0.10 <0.10 2,4+2,5-Dichlorophenol μg/l <0.20 <0.20 2,6-Dichlorophenol μg/l <0.10 <0.10 3,4-Dichlorophenol μg/l <0.10 <0.10 3,5-Dichlorophenol μg/l <0.10 <0.10 2,3,4-Trichlorophenol μg/l <0.10 <0.10 2,3,5-Trichlorophenol μg/l <0.10 <0.10 2,3,6-Trichlorophenol μg/l <0.10 <0.10 2,4,5-Trichlorophenol μg/l <0.10 <0.10 2,4,6-Trichlorophenol μg/l <0.10 <0.10 3,4,5-Trichlorophenol μg/l <0.10 <0.10 2,3,4,5-Tetrachlorophenol μg/l <0.10 <0.10 2,3,4,6-Tetrachlorophenol μg/l <0.10 <0.10 2,3,5,6-Tetrachlorophenol μg/l <0.10 <0.10 Pentachlorophenol μg/l <0.10 <0.10

Trichloromethane (chloroform) μg/l <0.30 <0.30

Tribromomethane (bromoform) μg/l <0.20 <0.20 Dibromochloromethane μg/l <0.10 <0.10 Bromodichloromethane μg/l <0.10 <0.10

All compounds, except AOX, were below the detection limit. However, AOX was also

detected in control water. The difference be tween the concentration in treated water and control water was not significantNIVA

18 Influent water, control water and treated wate rs in test cycle 3 were also sampled and analysed for Potential Bioaccumulating Subs tances (PBS) in acco rdance with OECD 117.

There were no difference in PBS-level between treated and control water. A value of 1.4 was

measure in both waters.

Treated waters in test cycle 4 were also sa mpled and analysed for free and total residual oxidants (TRO) by the DPD-colourimetric me thod (Hach method 8021 for free chlorine and Hach method 8167). The results show that it is not possible to de tect any production of

residual oxidants measured as free or to tal chlorine in the treated water.NIVA

19 

### Table 10. Concentration of free and total residual oxidants in treated water from Test 4 in influent water just prior to treatment, and control wate r and treated water immediately after treatment.

Unit Influent water Control water Treated water

Free chlorine Parallell 1 mg/l -0.04 0.03 0.02 Parallell 2 mg/l 0.00 0.01 0.01 Parallell 3 mg/l 0.05 0.02 0.02 Parallell 4 mg/l 0.02 - - Parallell 5 mg/l 0.00 - - Average± Standard

deviation 0.01±0.03 0.02±0.01 0.02±0.01

Total chlorine Parallell 1 mg/l 0.00 0.00 0.01 Parallell 2 mg/l 0.01 0.01 0.01 Parallell 3 mg/l 0.04 0.02 0.02 Parallell 4 mg/l 0.02 - - Parallell 5 mg/l 0.02 - - Average± Standard

deviation 0.02±0.01 0.01±0.01 0.01±0.01

3.2 Fulfillment of the biologi cal water quality criteria

3.3 Operational performance of the PureBallast system

A quality control was performed during each test by the site QA responsible person. Check points and operational parameters for docum enting sufficient operation of the treatment technology were included and monitored in this quality control, incl uding flow rate and voltage. Flow rate was read at the meter mount ed on the effluent pipe from the treatment technology. Voltage was read on the voltmet ers mounted on the treatment unit.

• Test 1: The flow rate in test 1 varied in the range of 182-220 m 3 /h during ballasting, and in the range of 191-205 m 3 /h during deballasting. The voltage was read to 150 V for each lamp. It was experienced some diffic ulties regarding the filtration. Frequent back-flushing was required.

• Test 2: The flow rate in test 2 varied in the range of 188-242 m 3 /h during ballasting, and in the range of 194-205 m 3 /h during deballasting. The voltage was read to 150 V for each lamp. The back-flushing of the filter was reduced due to change of screening

device.

• Test 3: The average flow rate in test 3 varied in the range of 240-251 m 3 /h, and in the range of 250-251 m 3 /h during deballasting. The volta ge was read to 150 V for each lamp.NIVA

20 • Test 4: The average flow rate in test 4 varied in the range of 245-254 m 3 /h, and in the range of 233-236 m 3 /h during deballasting. The volta ge was read to 150 V for each lamp.

3.4 Biocidal effects on organisms ≥ 50 μm in minimum diameter

3.5 Biocidal effects on organisms ≥ 10-50 μm in minimum diameter

3.6 Bactericidal effects

3.7 Ecotoxicological responses

The ecotoxicity of treated ballast water wa s tested once with undiluted (100 %) samples on three trophic levels using marine acute and ch ronic tests with untreated control water as

control. The tests and test results are summa rized below. The complete test reports are presented in the Appendixes.

Acute toxicity towards the juvenile turbot ( Scopthalmus maximus )

The acute toxicity of Treated Ballast Water toward s Turbot was tested in accordance with the draft procedure of Phil McWilliams (Terra Environmental Laboratory A/S) (ref 1). The procedure follows the general guidelines of OECD 203 "Fish, Acute toxicity test" (ref 2).

The testing of acute toxicity of Treated Ballast Water is required by the IMO G-8 guidelines for testing of treatment technology to be used aboard ships. The fish was exposed continuously for 96 hours with full water exchange every day. The juvenile turbot was approximately 12 weeks old at the start of the test and had a mean weight of 7.6 g. 20 fish were used in each aquarium with 20 l of medium. The temperature in the water varied in the range of 13.4-14.8 o C, and a light /dark cycle of 12:12 was used. The test water was taken directly from test tanks, after conditioning to test temperature

Treated Ballast Water did not exert any obs ervable effects on juvenile turbot.

The results of the test can be expressed as follows:

Parameter mg/l 95% confidence limits

LC 50 (96 h): >100 -

NOEC based on mortality: ≥ 100

LC 50 The concentration causing 50 % mortality

NOEC No observed effect concentration

Chronic toxicity towards the juvenile turbot ( Scopthalmus maximus )

The chronic toxicity of Treated Ballast Wate r towards Turbot was tested in accordance with the "OECD Guidelines for testing of chemicals" (No. 215; Fi sh, juvenile growth test), adapted for marine fish. (ref 1).NIVA

21 The testing of chronic toxicity of Treated Ballast Water is required by the IMO G-8 guidelines for testing of treatment technology to be used aboard ships. The fish was exposed continuously for 28 days with water exchange 3 times per week. The juvenile turbot was approximately 12 weeks old at the start of the test and had a weight in the range of 5 to 9 g. 20 fish were used in each a quarium with 20 l of medium. The temperature in the water was maintained in the range of 13,4 – 18,2 o C, and a continuous light was used. The test water was taken directly from test tanks, after condition to test temperature

There was no mortality observed in the test, and the exposed fish did not behave or look different

from control fish. Treated Ballast Water did not exert any significan t effects on the growth rate of juvenile turbot as determined using ANOVA statistics.

The results of the test can be summarised as follows:

Parameter Control Treated Ballast water

Average growth rate d -1 0.0063 0.0059

Reproductive toxicity to the marine crustacean Nitocra Spinipes

The reproductive toxicity of Treated Ba llast water to the marine crustacean Nitocra spinipes has been investigated. The test was performed according to Draft guideline for Danish Standard: ”Økotoksikologisk undersøgelse med krepsdyret Nitocra spinipe . Reproduksjonsmetode”. The test was performed as a Limit test with 100 % Treate d Ballast water and using non treated ballast water

as control water.

The test was performed with 20 replicate vessels w ith 1 gravid female in each vessel. The vessels were incubated for 14 days at 20 ° C. The number of alive offspring was then counted. There was no significant difference in numbe r of offspring in Treated Balla st water when compared to

control.

Test result: Average number of offspring Test Reproduction Standard Deviation

Control 21.9 9.4

Treated Ballast water 20.5 9.1

Growth inhibition of the marine alga Skeletonema costatum

The inhibitory effect of Treated Ballast water on the growth of the marine diatom Skeletonema

costatum , strain NIVA BAC1, has been investigate d. The test was performed according to ISO 10253: Marine algal growth inhibition test with Skeletonema costatum and Phaeodactylum

tricornutum.NIVA

22 Six replicate cultures of test al gae in untreated and treated cult ures were incubated on a shaking

table at 20 ± 2 ° C, under continuous illumination. Growth wa s monitored by daily counting of cell numbers using a Coulter Multisizer.

The growth rate in each culture was calculated from the increase in cell density from 0-48 and 0-72 hours exposure. The analysis of difference betwee n treatments was based on the exponential part of the growth curve (0-48 hours) . There was no significant differen ce in growth rates between the

untreated and treated Ballast water and the control sea water. Hence, for the treated Ballast water, the following results were obtained:

Effects on growth rate Concentration

%

48 h E

r C 50 1 >100

NOEC 2 ≥ 100

No growth inhibiting eff ect on the marine alga Skeletonema costatum was caused by the Ballast water treatment process.

Acute toxicity to the marine crustacean Acartia tonsa

The acute toxicity of Treated Ball ast water to the marine crustacean Acartia tonsa has been investigated. The test was performed according to ISO 14669: Determination of acute lethal toxicity to marine copepods ( Copepoda, Crustacea ). The test concentrations were in the range 32
to 100 % of Treated Ballast water.

The test was performed with four replicate vessels with 4-8 test animals for each test concentration and sixteen control replicate vessels. The vessels were incubated for 48 hours at 20 ± 1 ° C. Mortality was recorded after 24 and 48 hours.

A mortality of 27% was observed at the highest tested concentration (100%). It should be commented that high death-rate in the control can be a pr oblem with the test, as also experienced in the current test. This death-rate can give mislead ing results. No effects were observed in the test

with the other marine crustacean Nitocra spinipes . It is therefore doubtful that the effect observed can be attributed to toxicity.

Lack of mortality >50 % at any test concentration precludes estimation of LC 50 . Based on the results the following statement on the effect of Treated Ballast water on Acartia tonsa can be

made:

Concentration (%) 95 % confidence limits

48h LC 50 * >100 -

1 E

r C 50 : Concentration that causes 50 % reduction of the growth rate as compared to controls

2 NOEC: No Observable Effect Concen tration, i.e. the highest concentration that does not cause a significant inhibition of the growth rateNIVA

23 * LC = Lethal Concentration LC 50 represents the concentration that results in lethality of 50 % of the test animals.

Summary of toxicological tests

Based on the conducted acute and chronic toxicity tests it is concluded that ballast water treated with the presen t technology do not provide hazard or risk to the marine ecosystem

when released.