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.

I:\MEPC\59\2-8.doc

INTERNATIONAL MARITIME ORGANIZATION

IMO 

# E

MARINE ENVIRONMENT PROTECTION

COMMITTEE

59th session Agenda item 2 MEPC 59/2/8

16 December 2008 Original: ENGLISH

HARMFUL AQUATIC ORGANI SMS IN BALLAST WATER

Application for Basic Approval of the AquaTriComb  Ballast Water Treatment System

Submitted by Germany

SUMMARY

Executive summary: This document contains the non-conf idential information related to the application for Basic Approval of the AquaTriComb  Ballast Water Treatment System under the ìProcedure for approval of ballast water management systems that make use of Active Substances (G9)î adopted by resolution MEPC.169(57)

Strategic direction: 7.1

High-level action: 7.1.2

Planned output: 7.1.2.4

Action to be taken: Paragraph 5

Related documents: BWM/CONF/36; MEPC 57/21; BW M.2/Circ.13 and BWM.2/Circ.15

Introduction

1 Regulation D-3.2 of the Inte rnational Convention for th e Control and Management of Shipsí Ballast Water and Sediments stipulates that ballast water management systems that make use of Active Substances to comply with the Convention shall be approved by the Organization.

2 The ìProcedure for approval of ballast wa ter management systems that make use of Active Substances (G9)î stipulates th e required information (MEPC 57/21, annex 1, paragraph 4.2.1) and provisions for risk char acterization and analysis (MEPC 57/21, annex 1, section 5) which according to section 6 of the Procedure (G 9) should be evaluated by the Organization.

3 The receiving competent authority in German y has verified the application dossier and believes it satisfies the data require ments of Procedure (G9) adopted by resolution MEPC.169(57). In view of the fi ndings presented in the confidential and non-confidential parts of the application dossier , the national Administration does not deem anMEPC 59/2/8 - 2 -

I:\MEPC\59\2-8.doc application for Final Approval necessary at this time. The national Administration will submit such an application only if cl ear guidance is received or new facts emerge during the current national application process which ma ke such an application necessary.

4 In accordance with BWM.2/Circ.13 and BWM.2/Circ.15, Germany therefore submits the non-confidential part of the manufacturerís a pplication dossier in the annex. The complete dossier will be made available to the expert s of the GESAMP-BWWG with the understanding of

confidential treatment.

5 The Committee is invited to consider the pr oposal for approval and decide as appropriate.

***MEPC 59/2/8

I:\MEPC\59\2-8.doc ANNEX

NON-CONFIDENTIAL INFORMAT ION ON THE AquaTriComb  BALLAST WATER TREATMENT SYSTEM (GERMANY)

SUMMARY

Aquaworx ATC GmbH is applying for th e Basic Approval of the AquaTriComb TM ballast water treatment system (BWTS) at the International Maritime Organization.

The application of Aquaworx ATC GmbH for the Basic Approval of its ballast water treatment system AquaTriComb  is based on the technologically safe and environmentally sound approach to abstain from any preparations and avoidance of reactive intermediates with the potential to pose a risk to the environment a nd/or to human health. The AquaTriCombô BWTS is designed to purposely avoid reactive interm ediates like OHï radicals and consequently unwanted reaction products by using monochrom atic UV-light and low frequency ultrasound.

Based on definitions contained in the Methodolo gy for information gathering and the conduct of work of GESAMP (MEPC 58/2/7, annex 4), substan ces generated by the tr eatment process of the AquaTriComb  system are classified as: non Prepara tions are added and non- Active Substances are generated, Relevant Substance is defined as OHï radical and Other Ch emicals are defined as possible by-products.

To evaluate the impact of ballast water trea tment on the production of Other Substances water samples were analysed chemically before and after the treatment process. All THM, LHKW, HAA, Bromophenol, Acetonitril, AOX, EOX and TRO (at all 32 substances or substance groups)

measured before and after treatment were below Level of Quantification. One exemption being a Chloroform concentration in one sample before the treatment and is considered as an outlier. AOX could be measured in six samples before and after treatment at very low concentration at the Level of Quantification. Ni trite values showed a high va riability above the Level of Quantification, in samples before as well as in sa mples after the treatment process. To verify a significant assertion regarding th e variability of the results, a statistic test calculation was performed. The Mann-Withney U test statistic was us ed on the basis that data were not normally

distributed. Results of the test statistic showed the data from samples before and after treatment were significantly not different , whereby they belong to the sa me data population. Therefore, it can be concluded that Nitrite is not generated by the treatment process. All Nitrite values before and after treatment are within the German, EU, EPA and WHO drinking water regulations. Hence, and based on the measured data, no risk for human health and the environment can be diagnosed.

Under independent scientific ex amination the risk characterization and risk assessment was conducted regarding risks to the sa fety of ship, to human health and to the aquatic environment. Even if Nitrite values result from high background values originated in the test water and not from the treatment process, literature data of risk characterization and assessment for Nitrite was

conducted.

Analysis of the risk assessment demonstrates that no risks associated to the ship and its personnel neither to human health in genera l could be identified. Additionally it can be concluded that from this treatment process the magnitude of the haza rd and the probability of risk exposure to the aquatic environment is of no concern.MEPC 59/2/8

ANNEX Page 2

I:\MEPC\59\2-8.doc Based on analytic results and risk assessment, it can be concluded also that even if the precisely adjusted US (Ultrasonic oscill ation) and UV technology generate s OHï radicals by means of photons and cavitation bubbles, the concentration of OHï radical related Other Chemicals would be insignificantly low (not pres ent, or at non-measurable concentrations) and risks to human

health and to the environment can be demonstrably excluded.

Therefore, this application for Basic Approval is connected to the appeal to whether the AquaTriComb  BWTS proposal can be asse ssed according to Guidelines (G8) under the Ballast Water Management Convention.

1 INTRODUCTION

Aquaworx ATC GmbH is applying for th e Basic Approval of the AquaTriComb  Ballast Water Treatment System (BWTS) at the In ternational Maritim e Organization.

The application is connected to the appe al to examine whether the AquaTriComb  BWTS proposal can be assessed according to Guidelin es (G8) of the Ballast Water Management Convention, following the guidance and format provided in the revi sed Draft Methodology (MEPC 58/2/7). This is due to the fact that no Preparations are inte ntionally introduced or generated with the technology.

The AquaTriComb  BWTS is composed of pre- and after- treatment modules to disinfect aquatic species in the ballast water. The treatment system comprises mechanical filtration in combination with ultrasonic (US) oscillation for filter self-cleani ng, and ultraviolet-C (UV- C) radiation also in combination with ultrasound for self-cleaning of UV-C lamps.

Due to the highly efficient combination of US oscillation and UV radiation the AquaTriComb  technology can abstain from any Preparations or ch emical additives, which may pose a hazard to the population and/or to the environment.

Because the US and UV-C technology is precisel y adjusted to avoid the generation of OHï radicals, the concentration of th e OHï radical related Other Chemicals is that low that risks to

human health and to the environmen t can be demonstrably excluded.

UV-C and US are especially acknowledged in por table- and waste water management to produce no, or little disinfection by- products by themselves (Liu et al. 2006) and have been identified as the most environmental friendly and effective methods for ballast water treatment (Gavand

et al. 2007, Holm et al. 2008, Umweltbundesamt 2008).

All reactive intermediates generated by UV-C ra diation and ultrasound, and their potential reaction products during ballast water treatment occur naturally in seaw ater as products of sunlight induced photolytic activities (de Mora et al. 2000). In natural seawater reactive intermediates play an important role in th e degradation of dissolv ed organic matter (DOM; largely uncharacterized long-chain organic molecules) enhancing biodegradability by micro-organisms of such substances and thus are important for the oceanic carbon cycle (de Mora et al 2000). However, the A quaTriCombô Ballast Water Treatment System is designed to purposely avoid reactiv e intermediates and consequen tly unwanted reaction products by using monochromatic UV-light and low frequency ultrasound (Liu et al 2006, Holm

et al. 2008).MEPC 59/2/8

ANNEX

Page 3

I:\MEPC\59\2-8.doc The radical abandonment of any Preparations an d avoidance of reactive intermediates qualifies the AquaTriComb  technology as a highly effective and particularly environmentally sound ballast water treatment system.

1.1 Ultraviolet radiation

Ultraviolet technology is proving more and more successful and is being used as an acknowledged, cost-effective and re liable process. UV-C radiation as disinfection mean is well known for almost hundred years and is utilized widespread in the sector of disinfection of portable water, aquaculture waters, cooling water, hot water systems, ultra pure water, foodstuff industry, medicine, bottling of drinks, etc. It is used primarily wherever the microbiological requirements are particularly high, or where chem ical disinfectants are unacceptable for quality or environmental reasons. It was found that espe cially a wavelength of 2 54 nanometer (nm) has a strong performance for disinfection. The low-pressure mercury germicidal lamps, which are used for the UV-disinfection have their maximum pe rformance at a wavelength of 254 nm, and are similar to the absorption curve of the DNA.

The use of UV-radiation as disinf ection energy results in no change of the chemical compositions ph-value, temperature, salinity, taste, odour and co lour of the solution being treated, and safety issues related to the handling of Relevant and/or Other Chemicals are eliminated.

Ultraviolet disinfection requires a minimum dosage to be effective. The applied dosage is a function of the lamp intensity and the exposure ti me. These parameters are directly affected by equipment configuration, flow path of the wate r along the bank of lamps, the solids content, and the turbidity of the water to be disinfected. Compared to mid- or high-pressure lamps, low-pressure lamps can only operate with a lowe r performance, but they have a much better efficiency of about 45%. Using high- or mid-pr essure UV lamps, the operating costs will be higher. Due to these facts, for the disinfection appli cation of the treatment of ballast water low-pressure lamps are installed in the AquaTriComb  BWTS.

The low-pressure mercury vapour lamps consist of one hermetically closed discharge vessel. The gas inside is a vacuum seal against the outside. A voltage will be generated, which generates

an electrical field between the electrodes. If the accelerated volta ge is high enough, electrons will

leave the electrodes and will be accelerated in the el ectrical field. If they crash with the gas atoms they discharge their kinetic energy. The atom or mo lecule decreases to a lo wer energetic level, or to the basis level. This released energy will norma lly be emitted in kind of radiations, so-called ìquantsî. Due to the energy in the atom, merc ury is a very suitable gas for production of

UV-radiation.

It is a scientifically well-known fact that resistance of organisms like bacteria, viruses, moulds or fungal spores to ultraviolet radiation cannot be acquired. Most pathogenic germs are especially sensitive to UV-radiation. An impor tant advantage of physical disinf ection is that the sensitivity to UV still functions even when germs have already built up resistance to conventional

disinfection measures such as alcohol, antibiotics, chlorinate d chemicals, etc. A lot of conventional medical equipment and systems are now reaching the limits of their capabilities for disinfection and prevention when dealing with this problem.MEPC 59/2/8

ANNEX Page 4

I:\MEPC\59\2-8.doc To address all the mentioned needs, UV -radiation as part of the AquaTriComb  technology provides a powerful barrier to chemical contamin ants while simultaneously disinfecting to avoid the dispersal of invasive aquatic sp ecies by ballast water from ships.

The prime affect on organisms through short-wave and energetic UV-C radiation at 254 nm is based on a photo-chemical alterati on of the DNA. The UV-C radiation is absorbed by pyrimidins, which form the basic elements of the organic base of the DNA string. Th rough covalent bindings of two neighbouring bases like thymin, pyrimidin di mers are generated. These pyrimidin dimers avert replication of DNA helix strings and cause mutations wh ich result from unsuccessful DNA string repair mechanisms. Dimers are called ìbulky lesionsî: they disturb the helical configuration of DNA by means of partial decartelizati on resulting in reduced base matching ability. In addition, the UV radia tion may result in direct alte rations of the DNA morphology, for example, singular or double string fractures. Doubl e string fracture can not be repaired and lead to cell mortality. In general, UV radiation result in defects of DNA repair abilities.

The advantages of the application of UV -radiation as disinf ection process are:

- a purely physical disinfection process;

- no Preparations will be adde d nor intentionally produced;

- tried, tested and proven proce ss approved by the DVGW (DVGW 2001);

- water disinfected by UV-C radiation c onforms with German portable water regulations (TVO, BGBl 2001);

- no formation of health damaging by-products;

- no production or storage of chemicals on boa rd necessary, and no danger of over- or under-dosing;

- only naturally occurri ng processes are used;

- no change in physical parameters like pH -value, temperature, salinity, taste, odour, colour;

- the use of UV-C radiation does not in crease corrosion in ship compartments;

- low maintenance and operating costs of the UV reactor compartments; and

- low energy consumption needed by UV-C technology.

UV-C radiation as disinfection t echnology of ballast water gene rates no harmful environmental effects to the aquatic environment.

1.2 Ultrasonic oscillation

The special advantage of ultrasonic (US) oscillati on is the very fast and extremely high cleanness of the parts without any manual works to be carried out, and without any damage to the components. Also persistent impurities in, for example, borings and/or hollow spaces or at technically complicated parts will be rem oved by ultrasonic osci llation in seconds.MEPC 59/2/8

ANNEX

Page 5

I:\MEPC\59\2-8.doc Ultrasonic applications can be found, for example, in the avia tion industry, laboratory medicine, textile and plastic industry, print and semi-c onductor industry, surface engineering, etc. US cleaning and scouring is carried out with engine blocks, ball bearings, va lves, filters, nozzles, breathing apparatus, mechanical measurement and calibration device, analytic strainer, etc.

The germicidal effect of high frequency ultrasonic waves is assigned to ca vitation and its direct after-effects like shearing, pressure, local cale faction and generation of free radicals by sonoluminescence (electric discharge) during treatment in fluid media. It results in affection of the cell wall, the cell membrane and other functional cell elements (Save et al. 1994).

The advantages of the use of ultrasound are:

- ultrasound (US) processes are purely physical processes;

- the use of cavitation does not incr ease corrosion in ship compartments; - brushing or other means of cleaning is dispensable; - fast, highly efficient cleaning; - maintenance free, no personnel training needed; and - extremely low energy consumption.

1.3 Technology overview

Aquaworx ATC GmbH has developed a ballast wate r treatment system comprising pre- and after treatment modules. For the BWTS test plant the volume flow rate is designed at 250 m 3 /h. The capacity of the volume flow rate can be de signed by means of ali gned up-scaling of the modules. During ballast water intake and at pre- treatment stage the balla st water is filtered through three (3) identica lly constructed MicroSintFilter (MSF) modules to remove particles and organisms. Pre-filtration allows very effective removal of organisms.

Instead of so far applied 50 μm, the Aquaworx filter pore size equals 35 μm; filter material consists of a sintered high grade steel alloy. Th e automatically self-cleaning process of the filter modules is realized via permanent ultrasonic os cillation inside the fi lter tube. The ultrasound dissolves organic (bio film, aggl utinations) and inorganic (e.g., calcium carbonate) accumulations from the filter tube, and organic agglomerations (detritus particles, phytoplankton colonies, slime creating phytoplankton) in the ballast water ar e broken off, and organisms are pre-treated. By means of reversed flow rate the backflus h water is disposed off via separate piping.

After-treatment is processed in side the UV-reactor via UV-C low- pressure mercury germicidal lamps and low-frequency ultrasound. The applica tion of low-pressure UV-technology at 254 nm wavelength disinfects aquatic or ganisms very effectively.

The most important impact of the ultrasound is the highly effective and permanent cleaning of the UV-emitter through the ultrasonic transmi ssion along the full length of the UV-reactor, avoiding accumulations of bio film s or inorganic salts on the quart z-crystal tubes. In addition, organic particles and phytoplankton colonies are broken off and orga nisms are pre-treated. Additionally, inside the UV-reactor the organisms are lethally affected by UV-light and

ultrasound.MEPC 59/2/8

ANNEX Page 6

I:\MEPC\59\2-8.doc Figure 1: Aquaworx ballast water plant consisti ng of three identically constructed MSF-modules (MicroSintFilter) and UVC reactor (AquaTriLight)

As illustrated in Figure 1 the biological samples are taken in between the ballast water pump and the treatment plant (SP 01), in between the pr e- and after-treatment (SP 02), and behind the after-treatment (SP 04). At the sampling port SP 03 the quality of the backwash after disinfection can be measured during de-ballast operations.

The abiotic parameter as pH, co nductivity, temperature, turbidit y, oxygen content, flow rate volume and pressure are recorded during tests online via automatic sensors. The sensors are located at the ballast water feed pipe and behind the after-treatment. The piping diameter of the sampling points will be 1/8 of the ba llast water pipe diameter. All va lves are fitted with flanges,

and electrical-driven and manua l actuators. In addition to th e valve positions ìOpenî and ìCloseî, intermediate positions are possible.

In case of repair and maintenance or malfuncti on, the complete treatment plant can be by-passed with by-pass valves V01 and V03, and shut-off safety valves V02 and V012.

1.4 Conclusion: technology overview

The advantages of the AquaTriComb  ballast water treatment plant ATC-250 may be

summarized as follows:

- Radical abandonment of any Preparations and avoidance of reactive intermediates;

- Pore size of sinter filter (MSF modules) equals 35 μm;

- During de-ballast operations the UV reactor completely disinfects the discharge water by means of pre-treatment (filter and ultr asound) and after-treatment (ultraviolet radiation and ultrasound);MEPC 59/2/8

ANNEX

Page 7

I:\MEPC\59\2-8.doc - The application of ultrasound provides a hi gh clearance performance of the sinter tubes (filter) as well as the UV-tubes;

- High disinfection performance through combin ation of low-pressure UV-C radiation and ultrasound;

- Innovative technological optimization of filtration, ultrasound and UVC with extremely effective separation of partic les and sediments, and mortifying of organisms allows for low-energy consumption of only;

- As ultrasound effects are limited to the treatment plant components, cavitation effects on the shipsí piping and tank system are not possible, whereas the treatment plant components are designed to cope with cavitation;

- The application of low-pressure radiat ion and very low-frequency ultrasound with extremely short-termed ultrasound impact (< 1 s) generated, base d on analysis of treated water, none Other Chemical s during the treatment process;

- By the application of UV-disinfection mi cro-organisms can not develop resistance; therefore also resistant pathogens (e.g., ch lorine resistant) are inactivated by UV-C radiation; and

- UV-C radiation and ultrasound do not aff ect physical parameters like salinity or pH-values of the ballast water.

2 IDENTIFICATION OF PREPARATIO NS, ACTIVE SUBSTANCES, RELEVANT AND OTHER CHEMICALS (G9: 4.1)

2.1 Chemical identification

Based on the definitions of ìPreparationsî, ìActive Substancesî, ìRelevant Chemicalsî and

ìOther Chemicalsî as outlined in the revise d Methodology for information gathering and the conduct of work of GESAMP-BWWG (MEPC 58/2/ 7) substances used or generated by the AquaTriCombô BWTS are classified as follows:

2.1.1 Preparation means any commercial fo rmulation containing one or more Active Substances including any additives.

The AquaTriCombô BWTS does not utilize any ìPreparationî besides UV-light and low frequency ultrasound.

Not applicable.

2.1.2 Active Substance means a substance or organi sm, including a virus or a fungus that has a general or specific action on or against harm ful aquatic micro-organisms and pathogens.

The AquaTriCombô BWTS would produce ìPhotons î emitted by the low-pressure UV (LPUV) system, and ìCavitation Bubblesî generated by the low-frequency ultra-sound (LFUS) system. The photon and cavitation bubble are entirely responsib le for the inactivati on of harmful aquatic micro-organisms and pathogens present in ballast water.MEPC 59/2/8

ANNEX Page 8

I:\MEPC\59\2-8.doc With regard to the Methodology, both can not be classified as a s ubstance or organism, therefore they are defined as reactive physical processes, not as Active Substances. For this reason non-Active Substances are genera ted by the treatment process.

2.1.3 Relevant Chemical means transformation or reaction products that are produced during and after employment of the ballast water manage ment system in the ballast water or in the receiving environment and may be of concern to the shipís safe ty, aquatic environment and/or

human health.

The AquaTriCombô BWTS regards only the hydroxyl radical (OHï) as a ìRelevant Chemicalî. However, the amount of OHï potentially produ ced in the system is extremely small.

The OHï radical reacts at near ly diffusion controlled rates with any organic compound and exists

for about 10 -9 seconds.

Therefore, OHï is considered not resident in the treated ballast water and causes no harm to the environment upon discharge.

2.1.4 Other chemicals means reaction by-products th at could be formed in the ballast water upon treatment with Aq uaTriCombô BWTS.

During the process of OH-generation through phot olytic and sonolytic processes secondary

reactive intermediates and stable products can be formed. Most of the known stable products are non-toxic hydrocarbons. However, the formation of potential eco-toxicological Other Chemicals is partly known or can be inferred from appli cation of UV and US in portable and wastewater

treatment. Chemicals are specified under chapter 2.7.

2.2 Background to characterize Reactive physical processes, Relevant and Other

Chemicals

2.2.1 Reactive physical processes (G9, 2.1.1)

2.2.1.1 The photon

According to the wave/particle-duality light can be considered as energy propagating as a wave and as a stream of photons with the constant sp eed of light. A photon is defined as a quantum of electromagnetic energy at a gi ven frequency. Consequently, a photon possesses the more energy the shorter the wavelength. The energy characteri stic for a photon at a sp ecific wavelength is

determined as:

E = hc / λ (1)

Where E is energy in Joule, h is the Planck constant, c is the speed of light and λ is the wavelength in meters.

Ultraviolet light (UV) is part of natural sun light. Electromagnetic wa velength spectrum of UV (UV spectrum) is 100 ñ 400 nm bordered by x-ra y radiation (< 100 nm ) and visible light (400 ñ 700 nm). According to th e draft of ISO-standard on de termining solar irradiances (ISO-DIS-21348) the UV spectrum is subdivide d into UV-A (long wave, 315 ñ 400 nm), UV-B (medium wave, 280 ñ 315 nm), UV-C (100 ñ 280 nm), and vacuum UV (10 ñ 200 nm).MEPC 59/2/8

ANNEX

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I:\MEPC\59\2-8.doc UV-C can be used as a disinfectant against micr o-organisms and pathogen s including bacteria, viruses and phages (Hijnen et al. 2006). Therefore, UV-C is also referred to as germicidal light. Germicidal light is successfully applied in portable and wastewater management and the disinfection of water in aquaculture (Oppenl ‰nder 2003). The appli cability of UV in the treatment of ballast water of ships has been tested successfully in the laboratory and on the large scale (Welschmeyer et al. 2004).

The basic process of disinfection with UV light is photolysis. Photolysis is the decomposition of a molecule by light. The strength of the chemic al bond determines the photonic energy necessary for cleavage. Photonic energy of UV-C range s from 4.43 to 12.40 eV. A photon of UV254 nm contains 4.89 eV photonic energy.

For water disinfection two different UV tec hniques are widely used differing in their UV emission spectra. Middle Pres sure UV (MPUV) lamps emit light over a broad range of UV and visible light. Low-pressu re UV (LPUV) lamps used in the AquaTriCombô BWTS emit monochromatic light at the sp ecific wavelength of 254 nm (U V254nm) to specifically take advantage of the DNA damaging effect, and re duce the formation of potential by-products.

Absorption time for a photon by a molecule is in the range of nano- to picoseconds. It is generally accepted that photons leav e no residuals in water. Accord ingly, the use of UV for water disinfection and treatment (including ballast wa ter treatment) is consid ered a purely physical process producing negligible amounts of by-products (DVGW 2001, USEPA 2006, Umweltbundesamt 2008).

2.2.1.2 The cavitation bubble

Sound is a form of energy. Sound is generated by th e vibration of molecules in media (air, water, solid matter) and is travelling as a wa ve. The wavelength is defined as:

λ

= c/f (2)

where c is the speed of sound and f is the fre quency. Acoustic sound frequency range is 20 Hz s

inaudible and is referred to as ultrasound (US).

The uses of Ultrasound (US) within this large ra nge of frequencies are usually divided into two areas, i.e. low frequency US (20 to 100 kH z) and high frequency US (up to 10 mHz) (Mason & Lorimer 2002). Ultrasound has a broad range of application in industry, medicine and

water treatment. In water treatment mechanical and sonochemical properties of US are used to remove (micro-) organisms, chemical pollutants, to break down aggregations of particles and to clean filtration units. Ultrasound applied to wa ter has not enough energy to cause chemical reactions or mechanical disrupt ion itself (Mason & Lorimer 2002).

Chemical and mechanical properties rely on cavitation bubbles formed by US in liquids. Wave-like motion of US through water is charac terized as an oscillation of compression and rarefaction. If the negative pressure during rare faction is high enough the molecule structure of water is destroyed and gas f illed cavitation bubbles (CBs) are generated. Cavitation bubbles collapse violently releasing a shock wave and cr eate sheer stress causing mechanical disruption of material or organisms in close vicinity. Acco rding to this ìhot-spot-t heoryî drastic localized conditions are produced: temperatures of 2,000 to 5,000°K and pressures of 1,800 to 3,000 atm

occur in the CB.MEPC 59/2/8

ANNEX Page 10

I:\MEPC\59\2-8.doc As so, the CB itself acts as a micro-reactor tri ggering chemical reactions inside and outside the bubble. Hydrogen radicals (H·) and hydroxyl radica ls (OH·) can be generated inside the CB. Chemical reactions on the outside depend on the release of reactants formed inside the CB.

The lifetime of a CB created at low frequency in wa ter is in the range of μs to ms, where lifetime is mainly limited by collision. The CB itself is not resident in the water and consequently causes no harm upon release of treated ballast water.

2.2.2 Relevant chemical (OHï) (G9, 2.1.4)

We consider the hydroxyl radical (OHï) as th e only Relevant Chemical formed during the process of ballast water treatment by the Aqua TriCombô BWTS. Since OHï is highly reactive a number of chemicals could be produced in the treated ballast wa ter theoretically. The eco-toxicological analyses of most Othe r Chemicals are specified under chapter 2.7.

Formation of OHï can in genera l be initiated by both, UV-irradi ation and US. In the following, we are going to show by literature review and the results of chemical testing of ballast water treated by the AquaTriCombô BWTS, that:

.1 the AquaTriCombô BWTS is specifica lly designed to avoid the formation of OHï;

.2 therefore, amounts of OHï produced are extremely small (at all, and in comparison to other techniques); and

.3 consequently, potential formation of eco-toxicological relevant by-products is extremely low and concentrations are neglig ible and were not detectable in ballast water treated by the AquaTriCombô BWTS

2.2.3 Other chemicals

Oxidative degradation of organics by OHï is us ed in portable and wastewater disinfection. Very general, oxidative attack of OHï leads to the formation of the above-mentioned carbon centred radicals (RH2ï). The RH2ï immediately react with dissolved molecular oxygen producing a peroxyl radical (R-O-Oï). The R-O-Oï undergoes a vast variety of rearrangements and/or eliminations to form the final oxidation product (Rox). Final oxidation products are ketones, alcohols, aldehydes, molecular oxygen, aliphatic peroxides and H 2 O 2 depending on the aliphatic skeleton of the attacked organi c (Oppenl‰nder 2003). However, it ha s been demonstrated that the formation of carbonyls (e.g., aldehydes) at UV doses greater than 400 mJ/cm2 during wastewater and raw portable water disinfection is very low.

Due to the unpredictable nature of the compounds potentially formed from DOM degradation in natural seawater by OHï, we focus on co mpounds where formation is known and which potentially cause a risk to the environment or are regulated in water quality acts (e.g., trichloroacetic acid is a re gistered herbicide and regulated by the German portable water act (BGBI 2001). Of most environmental concern are H 2 O 2 , Trihalomethanes (THMs) and Haloacetic Acids (HAAs) as well as other organic halides (RX).

H 2 O 2 occurs naturally in seawater and is formed by photolytic and biological processes or is introduced by rainfall. H 2 O 2 is generated by the recombination of two OHï radicals:

OHï + OHï H 2 O 2MEPC 59/2/8

ANNEX Page 11

I:\MEPC\59\2-8.doc Natural decay follows photolytic and bi ological pathways . Half-life of H 2 O 2 in oligotrophic seawater estimated from decay rates under dark conditions is in the order of 16 ñ 224 h.

Sea water concentration of H 2 O 2 vary over a broad range (0.5 ñ 14 μg/l) depending on solar intensity, biological processes, rain input and the c oncentration of photolyt ic degradable DOM.

H 2 O 2 concentration in oligotrophic Atlantic surface water is abou t 100 nm (Bermuda triangle, Sargasso Sea) (Avery et al. 2004), while 400 nm H 2 O 2 have been measured in coastal waters.

The potential increase of H 2 O 2 concentration in different salinities during ballast water treatment by the AquaTriCombô BWTS is shown by wa ter analysis to be below the Level of Quantification.

Formation of THMs, HAAs and RX are common a nd regulated disinfection by-products (DBPs) during waste and portable water sanitation using chlorine or ch lorine species (Oppenl‰nder 2003, USEPA 2006).

Natural seawater contains organic halogens from anthropogenic input (chemical industry, chlorinated Seawater as cooling agent in power plants, portable water di sinfection) and natural sources (reactions of atmosphe ric organics with reactive hal ogens, production by algae, and sunlight induced photolysis of DOM). Concen trations of, e.g., THM and organic halides

measured in the North Sea and the Baltic are in the order of 0.2 ñ 15 ng/l- while composition depends on sampling site.

The potential increase of THM, HAA and RX con centration in different salinities during ballast water treatment by the AquaTriCombô BWTS is s hown by water analysis to be below the Level of Quantification.

Nitrite can be produced from N itrate by photolysis. The yield of the photoreaction of Nitrate to Nitrite is sensitive to both the dissolved inorganic and organic carbon content of natural water.

The nitrate concentration of untreated water infl uences directly the concentration of Nitrite produced as a consequence of UV and vacuum UV (VUV) irradiation. However, various doses of UV and vacuum UV irradiated natural organic ma tter were deemed non-cytotoxic. Similarly, a conducted Ames test showed no mutagenic res ponse after various does of both UV and VUV

irradiation.

Together with Ammonium (NH 4 +) and Nitrate (NO 3 -), Nitrite (NO 2 -) is the most common ionic (reactive) form of dissolved inor ganic nitrogen in aquatic ecosy stems. All these ions can be present naturally as a result of atmospheri c deposition, surface and groundwater runoff, dissolution of nitrogen-rich geological de posits, N2 fixation by certain prokaryotes (cyanobacteria with heterocysts, in particular), and biological degrada tion of organic matter.

2.3 Data on effects on aquatic plants, invert ebrates and fish, and other biota including sensitive and representative organisms (G9: 4.2.1.1)

This chapter has the purpose to evaluate the eff ects of Preparations and its components to the organisms and the environment when released w ith the treated ballast water or accidentally during the process.MEPC 59/2/8

ANNEX Page 12

I:\MEPC\59\2-8.doc AquaTriCombô BWTS applies Low-pressure UV (LPUV) lamps emitting monochromatic light at the specific wavelength of 254 nm (UV25 4nm). At this wavelength, specifically DNA is damaged and the formation of by-products is reduced.

Photons will not persist after the UVC exposure phase. Also OHï as Relevant Substance has a very short half-life and will not persist outsi de the BWTS system. Accordingly these two ìsubstancesî will be discussed below shortly w ith regard to their reaction mechanism and reactivity but not with rega rd to aquatic toxicity.

Nucleic acids and Nucleotides as components of DNA and RNA absorb in the wavelength spectrum of 200 ñ 400 nm with a distinct ma xima at 200 and 260 nm. Absorption of photon energy mediates the formation of various photol ytic products in DNA a nd RNA inhibiting gene expression (e.g., base dimers, strandñ and doubl e strand breaking, photohydrates). The most important, irreversible and lethal damage arises from the formation of pyrimidine base dimers, which block the translation and transcription of DNA and RNA, impeding replication and protein biosynthesis of micro-organisms (de Mora et al. 2000). The dimer formation is initiated by an exited state oxygen (Schreier et al. 2007). It has also been s hown that UVC (100 ñ 280 nm) causes photolytic inactivation of various proteins potentially e nhancing the germicidal effect (Chan et al. 2006 and references therein).

Absorption time for a photon by a molecule is in the range of nano- to pico-seconds. The use of UV for water disinfection and treatment (including ballast water treatment) is considered a purely physical process, limited to the time of radi ation exposure and producin g negligible amounts of by-products (DVGW 2001, USEPA 2006, Umweltbundesamt 2008). Hen ce no residuals of the photon are to be expected in the treated ballast water.

OH-radicals as ìRelevant Substanceî: Due to it s high oxidizing potential OHï is by far the most reactive transient species that is produced in this process. It reacts at a nearly diffusion controlled rate with most organic substances. Its life time of about 1 nanosecond rest ricts its process related appearance to the ballast water treatment phase. No increased concentrations outside the plant are expected.

2.3.1 Environmental toxicity of Other Chemicals ñ Nitrite

In the case of the AquaTriComb  BWTS Other Substances, which can theoretically be produced by the applied method as by-products within th e process, could be of potential concern. According to chemical analyses, none of the Ot her Substances, discussed in this dossier, are determined above Levels of Quantification (s ee discussion of results chapter 5.2) except for Nitrite, where values are above the LoQ both fo r measurements before and after the treatment process. Investigated Nitrite concentrations befo re and after treatment pr ocess are statistically proven not significantly different (see chapter 5.2), what means the Nitrite values are not generated by the treatment process. Nevertheless in the following chapters we discuss literature data on environmental toxicity for Nitrite.

2.3.1.1 Acute aquatic effects

The PAN database contains data to the Algae gr owth inhibition test fo r 72 hours for Tetraselmis chuii and states an EC50 of 159 mg/l. The validity to consider this test as an acute test, however, is doubtful, as several algae gene rations will have grown over this time. Other and more reliable acute data comprise EC50 values for crust acea (48 hours), ranging between 27 and 193 mg/l and fish (LC 50) ranging from 11.2 mg/L (Oncorhync hus mykiss) to 226 mg/l (Anguilla anguilla).MEPC 59/2/8

ANNEX Page 13

I:\MEPC\59\2-8.doc 2.3.1.2 Chronic aquatic toxicity

Data on chronic aquatic toxicity are scarce due to the very shor t life span of the compound which is quickly converted to other ni trogen species in an aquatic environment. Reliable data for chronic (NOEC) toxicity were not available.

2.3.1.3 Endocrine Disruption

An endocrine effect of specifica lly nitrate but also Nitrite is being disc ussed, but no data are available so far (Guillette, 2006). Up to now Sodi um Nitrite has not been listed as an endocrine substance in the EU priori tization list for endocrine disrupting substances.

2.3.1.4 Sediment toxicity

Under anoxic conditions it can be assumed that Nitrite is rapidly denitrified to, e.g., N2 O

.

2.3.1.5 Bioavailability, bioconcentra tion and food web effects

Accumulation of Nitrite has b een shown to depend on the environment and is higher in freshwater fish, where Nitrite and chloride ions compete for the active branchial Cl- uptake system. In seawater, bioaccumulation of NO2- in the plasma is much lower.

2.3.1.6 Overview over toxicological data of Nitrite

Table 1 shows literature data of acute (EC 50 , LC 50 ) toxicity for the organism groups algae,

crustacean and fish for Nitrite. Reliable data fo r chronic (NOEC) toxicity were not available.

Acute toxicity ñ EC 50 , LC 50 Organism

group Duration mg Nitrite /l Species Source

Algae 72 h 159 (EC 50 ) Tetraselmis chuii PAN Database

Crustacea 48 h 27,1 (EC 50 ) 193 (EC 50 ) Metapenaeus ensis

Penaeus monodon PAN Database

PAN Database

Fish 96 h 226 (LC 50 ) 130 (EC 50 ) 11,2 (LC 50 ) Anguilla anguilla

Danio rerio Oncorhynchus mykiss PAN Database

PAN Database
Kroupova et al. 2008

Table 1: Literature data of acute (EC 50 , LC 50 ) toxicity for the organism groups algae,

crustacean and fish to NitriteMEPC 59/2/8

ANNEX Page 14

I:\MEPC\59\2-8.doc 2.3.1.7 Evaluation of the PNEC

A PNEC of 0.01 mg/l has been determined by Bu rsch (2003) on the basis of chronic NOEC for three trophic levels 1 . This value has been suggested as the result of discussions by the competent authorities of the EU -member states. This level, however, has been critically

discussed as the level is strongly dependent on the ch loride concentration as well as other factors, and seemed unrealistically low.

The Environmental Quality Standards for Nitr ite have been proposed, based on data from Schwoerbel (1991) and addressing the differences in toxicity to aquatic organisms depending on the salinity of the ambient water:

0-3 mg Cl - /l ! 33 μg/l NO 2 - <3 ñ 7.5 mg Cl - /l ! 164 μg/l NO 2 - <7.5 - 15 mg Cl - /l ! 296 μg/l NO 2 - <15 - 30 mg Cl - /l ! 394 μg/l NO 2 - >30 mg Cl - /l ! 493 μg/l NO 2 -

2.4 Data on mammalian toxicity (G9: 4.2.1.2)

In the case of the AquaTriComb  BWTS Other Substances, which can theoretically be produced by the applied method as by-products within th e process, could be of potential concern. According to chemical analyses, none of the Ot her Substances, discussed in this report, are produced above Level of Quantific ation (see discussion of results chapter 5.2) except for Nitrite, where values are above the LoQ both for measurem ents before and after the treatment process. Investigated Nitrite concentrati ons before and after treatment pr ocess are statistically proven not significantly different, what means the Nitrite valu es are not generated by the treatment process. Nevertheless, in the following chapters we di scuss literature data on mammalian toxicity

conditions for Nitrite.

Information is taken from the background docum ent for ìdevelopment of WHO Guidelines for Drinking water quality: Nitrate and Nitrite in Drinking wate r, WHO/SDE/WSH/07.01/16,

WHO 2007î.

2.4.1 Acute toxicity

In a 13-week study in which Ni trite was given to rats in drinking-water, a dose-related hypertrophy of the adrenal zona glomerulosa was observed at all dose levels (100, 300, 1,000, or 3,000 mg of potassium Nitrite per litre). Increased methemoglobi n (metHb) levels were seen only in the highest dose group. FAO/WHO (1995) concluded that the NOEL in this study was 100 mg of potassium Nitrit e per litre (equivalent to 5 .4 mg/kg of body weight per day expressed as Nitrite ion), because the hypertrophy seen at this dose was not significantly different

from the controls.

2.4.2 Effects on skin and eye

No effects on skin or eyes have been reported.

1 Deriviation of the PNEC is documented in literature cited in Bursch (2003).MEPC 59/2/8

ANNEX Page 15

I:\MEPC\59\2-8.doc 2.4.3 Repeated dose toxicity

No repeated dose toxicity has been reported.

2.4.4 Chronic toxicity

One of the long-term effects of Nitrite reported in a variety of animal species is vitamin A deficiency; this is probably caused by the direct reaction of Nitrite with the vitamin. The most important effects reported in long-term animal studies were an increa se in metHb level and histopathological changes in the lungs and hear t in rats receiving Nitrite in drinking-water for two years. The LOAEL, which gave a metHb level of 5%, was 1000 mg of sodium Nitrite per litre; the NOEL was 100 mg of sodium Nitrite per litre, equivalent to 10 mg of sodium Nitrite per kilogram of body weight per day (or 6.7 mg/kg of body weight per da y expressed as Nitrite ion) (Speijers et al. , 1989).

2.4.5 Developmental and reproductive toxicity

Nitrite appeared to cause fetotoxicity in rats at drinking-water concentra tions equivalent to 200 and 300 mg of sodium Nitrite per kilogram of body weight per day, cau sing increased maternal metHb levels. However, after similar doses in f eed in other studies, no embryotoxic effects were observed in rats. In a reproductive toxicity study in guinea pigs at dose levels of 0, 50, or 60 mg of sodium Nitrite per kilogram of body weight per day given by subcutan eous injection, fetal death followed by abortion occurred at the highe st dose level. Teratogenic effects were not observed in reported studies in mice and rats (Speijers et al. , 1989; FAO/WHO, 1996).

2.4.6 Carcinogenicity and mutagenicity

Nitrite is mutagenic and can cause morphological transformations in in vitro systems; mutagenic activity was also found in a combined in vivoñin vitro experiment with Syrian hamsters. The

results of in vivo experiments were controversial (Speijers et al. , 1989; FAO/WHO, 1996).

At lower Nitrite levels, tumour incidence potentially induced by N-nitroso compounds, resembled those of control groups treated with the nitrosatable compound only. On the basis of adequately performed and reported studies, it may be concluded that Nitrite itself is not carcinogenic to animals (Speijers et al. , 1989; FAO/WHO, 1996, 2003a).

2.4.7 Toxicokinetics

No data on toxicokinetics are available.

2.5 Data on environmental fate and effect under aerobic and anaerobic conditions (G9: 4.2.1.3)

In the case of the AquaTriComb TM BWTS Other Substances, wh ich can theoretically be produced by the applied method as by-products within the process, could be of potential concern. According to chemical analyses, none of the Ot her Substances, discussed in this report, are produced above Level of Quantificat ion (see discussion of results, ch apter 5.2) except for Nitrite, where values are above the LoQ both for measurem ents before and after the treatment process. Investigated Nitrite concentrations before and af ter the treatment process are statistically proven, not significantly different, which means the Nitr ite values are not gene rated by the treatment process. Nevertheless, in the fo llowing chapters we discuss literat ure data on environmental fate

and effect for Nitrite.MEPC 59/2/8

ANNEX Page 16

I:\MEPC\59\2-8.doc 2.5.1 Nitrite

Together with Ammonium (NH 4 +) and Nitrate (NO 3 -), Nitrite (NO 2 -) is the most common ionic (reactive) form of dissolved inor ganic nitrogen in aquatic ecosy stems. All these ions can be present naturally as a result of atmospheri c deposition, surface and groundwater runoff, dissolution of nitrogen-rich geological de posits, N2 fixation by certain prokaryotes (cyanobacteria with heterocysts, in particular), and biological degradation of organic matter. Nitrite is an important nutrient within th e nitrogen cycle which is depicted below.

Figure 2: The biological nitrogen cycle. Figures in parenthese s denote the nitrogen

oxidation state ANAMMOX: anaerobic oxidation of ammonia with Nitrite (redrawn from Richardson (2000)).

2.5.1.1 Modes of degradation

Typical concentration ranges of Ni trite in seawater have been stated to be 0.01 to 5 μg at/l (corresponding to 0.46 to 230 μg NO 2 -/l. Nitrite as an essential el ement of the nitrogen cycle is

involved in a number of aerobi c and anaerobic processes. Under oxic conditions, e.g., in the water column or in the oxidized upper sediment layer, chemolithotrophic bacteria, that build organic matter from CO 2 , use ammonia and Nitrite (NO 2 -) as electron donors in a process called nitrification, which yields energy for the orga nisms. The complete oxidation from ammonia to nitrate (NO 3 -) is carried out by two differe nt groups of ìnitrifying bact eriaî that act in concert: Nitrosomonas and Nitrobacter. Nitrogen compounds are removed from the system by denitrification, which occurs under anaerobic c onditions: Nitrate and N itrite act as electron acceptors during anaerobic respiration. The nitrogen-compounds are finally reduced to gaseous compounds N 2 or N 2O . This process is the main process by which gaseous N 2 is formed biologically.MEPC 59/2/8

ANNEX Page 17

I:\MEPC\59\2-8.doc 2.5.1.2 Bioaccumulation and persistence

Nitrite under normal ìstableî environmen tal conditions is a transient inorganic nitrogen-compound which is quickly transforme d by oxidation or reduction processes (see 2.5.1). Under conditions of unbalanced nitrification, due to anthropogenic activit ies, problems may be created and are described in chapter 2.3.1. Clark et al. (2007) showed with samples from the North Atlantic Ocean, that Nitrite turnover ra tes due to ammonium and Nitrite oxidation can be highly variable in oligotrophi c waters, ranging from < 1 day to > 10 days. Influential factors that regulate the activity of nitrifying bacteria a nd, thus, the nitrification rates in general, could comprise ammonium-availability, pH, temperatur e, oxygen concentration, competition for NH4, and organic carbon availability. Hen ce, concrete assumptions on the persistence of Nitrite are not possible, other than it is very little persistent when comp ared to non-nutrient compounds.

Nitrite can accumulate in freshwater fish, as it competes with chloride ions for the active Cl-uptake mechanisms. Since the concentration of ch loride ions in freshwater in low, presence of

Nitrite can lead to NO2-accumula tion. In seawater, the much higher chloride concentration limits the uptake, and hence leads to low uptake of Nitrites from surrounding water. Kroupova et al. (2008) found elevated Nitrite levels in the plasma of fish (rainbow trout) exposed to concentrations of 1 mg/l NO 2 - and greater.

2.5.1.3 Reaction with organic matter

Nitrite does not react with organi c matter in a relevant way.

2.5.1.4 Potential physical effects on wildlife and benthic habitats

No physical effects on wildlife a nd benthic habitats are assumed.

2.5.1.5 Potential residues in seafood

Of potential concern with regard to contamin ation of food with nitrogen compounds is the

formation of nitrosamines which are formed in smoked fish products an d in the human stomach as the result of the simultaneous presence of secondary amines and Nitrite. Most concern, however, seems to be directed towards the use of nitrogen compounds as preservatives, rather than their uptake by humans from fish whic h had accumulated Nitrites. As the knowledge on Nitrite accumulation is fairly new and results so far have been very variable, more research seems to be necessary in this respect.MEPC 59/2/8

ANNEX Page 18

I:\MEPC\59\2-8.doc 2.6 Physical and chemical properties for the Preparations, Active Substances and treated ballast water (G9: 4.2.1.4)

Physical/Chemical Property Active

Substance:

Photon Relevant

Chemical:

OH-Radical Other

Chemical:

Nitrite Treated

Ballast

water

CAS Nr. n.a. 3352-57-6 14797-65-0

Molecular weight [g/mol] n.a. 17.00734 46

Melting point (°C)

Boiling point (°C)

Density

Vapour pressure

Water solubility

Dissociation constant (pK

a )

Henryís law constant

Log K OW Oxidation potential

pH in solution basic

Corrosivity to material

Reactivity to container

material

Autoignition temperature (°C)

Explosive properties

Oxidizing properties

Surface tension

Viscosity

(Thermal) stability and breakdown product

Other physical and chemical properties

Table 2: Physical and chemical properties fo r Physical reactive pro cess (Photon), Relevant Chemical (OH Radical) and Other Chemicals (n.a .: not available, grey column 2 and 3: due to the transient existence, no data are available, column 5: data not available, column 4 Nitrite: no information possible, as it is an ion, all information would refer to the salt, whereby the diffe rent compounds would differ strongly depending on the cation, Data from Rippen (2005))MEPC 59/2/8

ANNEX Page 19

I:\MEPC\59\2-8.doc 2.7 Analytical methods at environmenta lly relevant concentrations (G9: 4.2.1.5)

In chapter 2.2 the photon (UV-C radiatio n) and cavitation bubbles generated by the AquaTriComb  BWTS have been identified as respon sible for the inactivation of organisms, which in turn may result in OH-radicals. OH-ra dicals are extremely shor t-lived measured in nanoseconds, thus detection by co nventional analytics is not f easible. Reactive effects of OH-radicals may result in reactive by-products li ke ozone, hydrogen peroxide or hyper bromate. Other possible OH-radicals related by-products are THMs, LHKWs, HAAs, Acetonitrils or Bromophenols. These by-products have been analysed by three laboratories.MEPC 59/2/8ANNEXPage 20I:\MEPC\59\2-8.docBasic ApprovalGloEnPatrolMEPC 57/2/4GESAMP-ListMEPC 58/2/7UBA-ListBasic ApprovalAquaTriCombFormulaAnalysedAOX AOXAOX AOX YesEOXEOX EOX YesNitrite (NO2)-Nitrite (NO2)-NO2YesCl2Cl2H202YesHydrogen peroxide Hydrogen peroxideH202YesOzone OzoneO3YesHyperbromate Hyperbromate BrO- YesChloroform Chloroform Chloroform Chloroform CHCl3YesBromoform Bromoform Bromoform Bromoform CHBr3YesDibromochloromethan Dibromochloromethan Dibromochloromethan Dibromochloromethan CHBr2Cl YesDichlorobromomethan Dichlorobromomethan Dichlorobromomethan Dichlorobromomethan CHCl2Br YesMonochloracetic acidMonochloracetic acid C2H3ClO2YesDichloroacetic acidDichloroacetic acid C2H2Cl2O2YesTrichloroacetic acidTrichloroacetic acid C2HCl3O2YesMonobromoacetic acid Monobromoacetic acid Monobromoacetic acid Monobromoacetic acid C2H3BrO2YesDibromoacetic acid Dibromoacetic acid Dibromoacetic acid C2H2Br2O2YesTribromoacetic acid Tribromoacetic acid Tribromoacetic acid C2HBr3O2YesBromochloroacetic acidBromochloroacetic acidBromochloroacetic acid C2H2BrClO2YesDibromochloroacetic acid Dibromochloroacetic acidDibromochloroacetic acid C2HBr2ClO2YesBromodichloroacetic acidBromodichloroacetic acid C2HBrCl2O2YesMEPC 59/2/8ANNEXPage 21I:\MEPC\59\2-8.docBasic ApprovalGloEnPatrolMEPC 57/2/4GESAMP-ListMEPC 58/2/7UBA-ListBasic ApprovalAquaTriCombFormulaAnalysed1,2- dibromoethane1,2- dibromoethane C2HBr2Yes1,2,3-trichloropropane1,2,3-trichloropropane C3HCl3Yes2-chlorotoluene2-chlorotoluene C7H7Cl Yes4-chlorotoluene4-chlorotoluene C7H7Cl Yes1,2-dibromo-3-chloropropane1,2-dibromo-3-chloropropane1,2-dibromo-3-chloropropaneC3H5Br2ClYes1,2,3-tribromobenzene1,2,3-tribromobenzene C6H3Br3Yes1,1,1-Trichlorethan1,1,1-Trichlorethan C2H3Cl3YesCarbon tetrachlorideCarbon tetrachloride CCl4YesTetrachlorethenTetrachlorethen C2Cl4YesMonobromoacetonitrile Monobromoacetonitrile Monobromoacetonitrile C2H2BrN YesDibromoacetonitrileDibromoacetonitrile C2HBr2N YesBromochloroacetonitrileBromochloroacetonitrile C2Br3N Yes2,4-dibromophenol2,4-dibromophenol C6H4Br2O Yes2,6-dibromophenol2,6-dibromophenol C6H4Br2O Yes2,4,6-tribromophenol 2,4,6-tribromophenol 2,4,6-tribromophenol C6H3Br3O YesTable 3: List of substances identified aspossible by-products andanalysed by Aquaworx.MEPC 59/2/8

ANNEX Page 22

I:\MEPC\59\2-8.doc In Table 3 three different lists identify disinfection by-products (column 1 to 3), non-conformity between the lists is marked ìyellowî:

- Column 1 as given in MEPC 57/2/4 is a combination of tec hnologies similar to
AquaTriComb  BWTS;

- Column 2 according to GESAMT Ball ast Water Group, MEPC 58/2/7; and

- Column 3 according to recommended list of possible by-products as given by German Federal Environment Agency (Umw eltbundesamt Deutschland, UBA).

Water samples were analysed regarding the subs tances from all lists as shown in column 4.

The following Table 4 shows Other Chemicals including Levels of Quantification and

identification numbers of the standardized methods analysed by three laboratories.MEPC 59/2/8

ANNEX Page 23

I:\MEPC\59\2-8.doc Parameter Level of Quantification Methods

Nitrite (NO 2 ) 0,007 mg/l DIN EN 26 777 1993-04 AOX 0,02 mg/l DIN EN 1485: 1996-11 EOX 0,002 mg/l DIN 38141-S17 1989-11

THM:

Chloroform

Bromoform

Dibromochloromethan

Dichlorobromomethan 0,02 μg/l EN ISO 10301 1997-04

LHKW 1,2- dibromoethane 1,2,3-trichloropropane

2-chlorotoluene

4-chlorotoluene 1,2-dibromo-3-chloropropane 1,2,3-tribromobenzene 1,1,1-Trichlorethan

Carbon tetrachloride

Tetrachlorethen 0,02 μg/l EN ISO 10301 1997-04

HAA:

Monochloracetic acid

Dichloroacetic acid

Trichloroacetic acid

Monobromoacetic acid

Dibromoacetic acid

Tribromoacetic acid

Bromochloroacetic acid

Dibromochloroacetic acid

Bromodichloroacetic acid 1,0 μg/l DIN 38407 F25

Bromophenols: 2,4-Dibromophenol 2,6-Dibromophenol 0,01 μg/l GC-MS after derivatization and

extraction

2,4,6-Tribromophenol 0,04 μg/l GC-MS after derivatization and

extraction

Acetonitrils:

Monobromoacetonitril

Dibromoacetonitril

Bromchloroacetonitril 1,0 μg/l US EPA Method 551.1

TRO: Cl 2 , H 2 O 2, O 3, BrO All substances: 0,05 mg/l EN ISO 7393-1 2000-04 (G4-2)

Table 4: Possible by-products including Levels of Quantification and cer tified methods analysed by three certified laboratories.MEPC 59/2/8

ANNEX Page 24

I:\MEPC\59\2-8.doc Certified standards are followed with regard to th e samples at the beginning of every analytical run. Every method is also include d in periodical participation on extern laboratory tests except for Bromophenols, see below.

Levels of quantification

According to the report of the seventh meeting of the GESAMP-Ballast Water Working Group in annex 5 (MEPC 58/2/8) Levels of Quantifica tion (LoQ) for THMs and HAAs for surface waters are recommended as 0.01 ñ 0.05 μg/l. The laboratories involved in our analysis were able to reach LoQs for THM, LHKW and Bromophenol s of 0.02 μg/l, which meets the recommended range. In the case of the HAAs, expe rts of the laboratories have spen t considerable effort to find methods or laboratories, which are able to m eet the recommended LoQ. The effort to find a solution also included the attempt, to establish and optimize a standardized analytical method regarding this special de mand. As the best result a laboratory was able to reach a LoQ for HAAs with 1 μg/l. Experts of the labo ratory explain the background of the applied analytical method and the reason why the LoQ of 0.01 ñ 0.05 μg/l could not be reache d (see below analyse method of HAAs).

Analyse methods

The analyses of AOX/EOX have been carri ed out according to DIN EN 1485: 1996-11

and DIN 38141-S17 1989-11 with a coulometer of company Umcat. The parameters are so called summation parameter. They detect certain matters in their complexity and allow for interpretations which are important with regard to the advanced analysis strategy and to the plausibility control. As to AOX and EOX it is the sum of the halogenated absorbable and extractable compounds within the probe. The measur es to assure quality for such analyses are defined by DIN. That is besides the blank value of the instrument, the blank value is carried out for the analysis including a regul ar calibration by means of external standards. For the day-to-day

control a control standard is an alysed and control readings sh all detect methodical problems. Further for AOX regular participation of in terlaboratory tests is carried out.

Lightly volatile halogenated hydroc arbons and Trihalogenmethans ha ve been analysed according to DIN EN ISO 10301 1997-04 with a gas-phase chromatograph with mass spectrometric detection and headspace sampler of Messrs. Agile nt. The advantage of detection with a mass spectrometer is the definite identification and classification of the detected substance. The application of the SIM method, which account s for the specific masses of the detected substances only, allows for very small LoQ. For these parameters the la boratory also regularly participates in interl aboratory tests. The qua lity assurance methods for these analyses are determined by DIN. That is besides the blank valu e of the instrument, the blank value is carried out for the analysis including a regular calibration by means of extern al standards. For the day-to-day control a control standard is analysed and cont rol readings shall dete ct methodical problems.

Nitrite is analysed according to DIN EN 26 777 1993-04 with a photomet ric procedure. At a specific wavelength the colour reaction is detect ed with a reagent. The procedure for quality assurance is according to DIN and includes a daily calibration, a blind valu e as well as control

standards which are documen ted on control charts.

The analysis of ozone, hydrogen peroxi de and hyper bromate has been done in situ with the

method for free chlorine as the sum of the existi ng oxidants in water. A mobile photometer has been applied which allows for the measurement of the colour intensity as direct reaction withMEPC 59/2/8

ANNEX Page 25

I:\MEPC\59\2-8.doc N,N-Diethyl-1,4-phenylendiamin and formati on of a reddish compound at ph 6.2 to 6.5 according to EN ISO 7393-1; 2000-04 (G4-2).

Bromophenols were analysed with a gas-phase ch romatograph with mass spectrometric detection and autosampler of the company Agilent. Analysis is done after derivatiza tion as methyl ester. The advantage of detection via mass spectrometer is to found in the defin itely identification and

in the clear classification of th e investigated substances. By means of using the SIM method, which considers only specific masses of analysed substances, very low LoQs can be achieved. For these parameters the laboratory can not take part in interlabor atory tests (ring tests), because these parameters are until now not implemented in ring tests. Quality control measures for this analysis include beside blind values of the instrument, bli nd values for the method also permanent calibration with external standards. For the daily control a control standard is analysed.

Difficulties arose to find a laboratory to analyse halogenated Acetonitrils. Analysis of these substances is very seldom asked for, and theref ore laboratories have rare ly established respective certified analytic methods. The laboratory an alysed Acetonitrils acc ording to the US EPA method 551.1. With regard to the sound solubility of the Acetonitrils the salt sodium sulphate was added (shifting of the solution equilibriu m) and subsequently extracted with MTBE.

The extract will be concentrated and then anal ysed with GC/MS. All three Acetonitrils are measured with a LoQ of 1 μg/l.

Determination of HAA was performed following the guideline EN ISO 23631:2006. The analytical range was within 0.5-10 μg/l according to the standard solutions employed, depending on the matrix of the samples, respectiv ely. Our estimated limit of quantita tion was 1 μg/l, which is appropriate for the anal ytical range stated above. Descri ption of method and analytical constraints: 200 ml of aqueous te st solution is adjusted to pH 1 and extracted after addition of 20g sodium chloride with 20 ml MTBE. Subs equently, the organic extract is removed and

concentrated to 1 ml. After deri vatization of the anal yte with diazomethan, concentrations were assessed by means of GC-ECD. However, si nce the compounds under study are very polar

extraction is rather difficult and will be eventu ally not complete. Furthermore, it will not be possible to decrease the limit of quantitation by increasing the sample volume thus leading to higher concentrations of the analyt e in the organic extract. This is due to the fact the matrix will

also be concentrated in this way, leading to unfavourable di sturbances of the detection by GC-ECD.

2.8 Quality control/quality assurance (G9: 4.2.4)

For the laboratories the Analytic Quality Assura nce (AQS) is of utmost importance. It is the prime aim to deliver reliable analytic results of defined quality. Therefor e, one of their main targets is the quantitative asse ssment of all applied analys e methods, if necessary the improvement of these methods, and to ensure an d to document the achi eved quality throughout the analytic routine.

To meet this challenging target a quality manage ment system (QM) is implemented. This system governs all quality-related routin es and provides the pre-condi tions for continuous quality improvement according to DIN EN ISO 17025 and ìGood Laboratory Practiceî (GLP).

To comply with the high demands regarding analyses in the water pertaining enforcement, the existing AQS measures have been al igned to the ìRahmenempfehlungen der L‰nderarbeitsgemeinschaft Wasser (LAWA) f ̧r die Qualit‰tssicherung bei Wasser-,MEPC 59/2/8

ANNEX Page 26

I:\MEPC\59\2-8.doc Abwasser-und Schlammuntersuchungenî (basic re commendations of the working group water for the quality assurance of wate r, waste water and sludge analys es). On this basis IBEN is accredited according to DIN EN ISO 17025 and hol ds several other concessions for legally regulated sectors. That means that in addition to internal quality assurance external controls in form of mandatory and periodic interlaboratory tests have to be carried out. This in turn guarantees the permanent control of services. Part icipation in interlaboratory tests is obligatory by means of the accreditation, that is, that these tests have to be performed to maintain the

accreditation.

3 USE OF ACTIVE SUBSTANCE(S)

Although the AquaTriComb  does abstain from any Preparations the highly efficient

combination of ultraviolet radiation and ultrason ic oscillation generate s photons and cavitation bubbles. Nor photon or cavitation bubbles are residual in the treated ballast water and thus have no toxicological effects upon disc harge into the environment.

The OH radical, which can be ge nerated by photons and cavitati on bubbles reacts at nearly

diffusion controlled rate s with organic compounds and exists for about 10-9 sec onds. Therefore,

OHï is considered not resident in the treated ba llast water and causes no harm to the environment upon discharge.

The OH radical related by-products are discussed above in chapter 2, and data on analysed water samples showed none of the investigated substa nces are generated by the treatment process regarding to the applied Levels of Quantitation. For this risks to human health and/or to the environment can be demonstrably excluded.

3.1 Operational overview and process description

The AquaTriComb  BWTS will be installed in line behind the ballast water pumps with a bypass pathway. Both ballast water intake flows, as well as de-ballasting flows, will be directed via the whole treatment system.

Two of the three MicroSintFilter modules are pe rmanently active in the filtrati on process, the third module is in cleaning and stand-by mode.

Pressure difference in front of and behind the treatment system will be approximately 1 bar, depending on the contamination of the water. From the system the ballast water is directed into

the ballast tanks and via the ballast wate r pump(s) into the sea, respectively.

3.2 System monitoring and control, crew interaction

The AquaTriComb  treatment plant is fully automated and completely self-contained with electronic steering, switchboard, and control panel including emergency shut-off device. The plant can be readily pre-mounted in accordan ce to the shipís specifics for connection to the shipboard power supply system.

The monitoring and control devices include flow meters, pressure transmitter, electronically

driven actuator valves with manual setting option, and sample ports. Also operating hours

counter and malfunction writer will be installed (data logging).MEPC 59/2/8

ANNEX Page 27

I:\MEPC\59\2-8.doc Depending on the grade of automati on of the existing ballast water system, typically the plant requires no additional action by the crew or alterations of current ballast water procedures. If any, minor modifications to ballasting procedures are ship specific a nd will be carefully considered during system engineering and installation.

3.3 System requirements

Space requirements

The treatment plant is designe d as a modular system ñ all single filter modules and the UVC-reactor can be installed independently. For th is reason in the case of refitting ships with AquaTriComb  BWTS the existing space can be optimally used.

As the MSF modules as well as the UV-reactor are vertically installed sufficient top space has to be foreseen for exchange of filter m odule and US oscillators, respectively.

Personnel requirements

The treatment plant is designed to need min imum input from the crew. The crew member performs ballasting and de-ballasting pr ocedures as normal and the AquaTriComb  starts operations after activation of switch-on device. Fu rther, Aquaworx is offering a training unit to the key shipboard personnel to get familiar with the system.

Maintenance requirements

The ultrasonic tube oscillator s need to be exchanged at th e earliest after approximately 16,000 and 20,000 operating hours, the low-pressure me rcury germicidal lamps after about 12,000 operating hours. All other system components do not need any extra maintenance and will be integrated into the routinely applied shipboard plant maintena nce schedule. Depending on the ship and the shipís trade a maintenance contract will be signed considering periodic reviews and other specifics.

Handling of waste and replaced pa rts of the treatment system

During the treatment process no liquid or solid wa ste appears. Only in the case of completed operating hours, or broken, or out-of-order UVC lamp s, the glass tubes of the lamps have to be changed and disposed. In case of broken glass no hazard appears to the ship crew, because the mercury is as amalgam accumulated as a thin laye r on a special spot the inner glass tube. Broken glass can be disposed on board similar to other glass waste. In case a whole lamp be removed from the reactor, the lamp should be kept on board for recycling by Aquaworx or by the lamp

manufacturer.

3.4 System locations

The AquaTriComb  BWTS can be located in any suitable and convenient location dependent on the shipís design. The naval architects and mari ne engineers of Aquaworx will consider and discuss with the client to keep engineering for th e integration of the BWTS at a low level, to not interfere with normal cargo operatio ns and required crew activiti es, and to look to placement opportunities next to the ballast wate r pumps to minimize the piping length.MEPC 59/2/8

ANNEX Page 28

I:\MEPC\59\2-8.doc 3.5 Operational safety features and alarms

As not otherwise permissible and acceptable in the shipping indus try, all system components will be of maritime standard type and will have the appropriate approval of the classification society and/or relevant regulatory authority.

No rotating parts or other physical hazar ds are originated by the AquaTriComb  BWTS; hazards

from chemicals or other Active Substances are excluded. At least the following alarms will be indicated at th e BWTS switchboard:

- UV-reactor operational readiness after appr oximately 5 minutes warm-up phase after switch on (red to green);

- BWTS in by-pass mode (red); and

- General malfunction (red)

These alarms may also be indicated at the Engi ne Control Room (ECR), the Cargo Control Room (CCR) and at the bridge, or a common alarm will be installed there.

An emergency shut-off device will be inst alled at the BWTS and at the ECR.

3.6 Procedures and management measures

The operation of the AquaTriComb  BWTS is to be integrated into the Ship Safety Manual (SMS) as an integral part of the ISM 2 documentation.

That is, a procedure has to be developed based on the AquaTriComb  BWTS Guidelines for the proper operation of the treatment plant. This procedure also describes the service and maintenance intervals and regular preventive main tenance measures (routine check of sea chest, piping corrosion and relate d shipboard routines).

3.7 Evaluation of Management Measures

Preventing exposures and emergency situations star ts with proper engineering and installation of the AquaTriComb  BWTS on board the ship. The mana gement measures discussed above consider location, training and hand ling of the plant. In connection with the risk assessment these measures are considered to be adequate to av oid uncontrolled or emerge ncy situations and to operate the ballast water treatment plant in a safe manner.

4 MATERIAL SAFETY DAT A SHEETS (G9:4.2.7)

Since Material Safety Data Sheets (MSDS) for th e Relevant Chemical OH radical does not exist, MSDS of several investigated Other Chemicals can be found in Annex C.

2 International Safety Management (ISM) Code: International Code for the Safe Operation of ships and for Pollution Prevention, SOLAS chapter IX.MEPC 59/2/8

ANNEX Page 29

I:\MEPC\59\2-8.doc 5 RISK CHARACTERIZATION

In this chapter, the information, displaye d in the foregoing chap ters concerning hazard identification, exposure assessment and dose-res ponse assessment are summarized in terms of

risk assessment.

5.1 Screening for persistence, bioa ccumulation and toxicity (G9:5.1)

Photons and cavitation bubbles as well as hydrogen radicals as Relevant Substance have a very low life span of less than nanoseconds (see chapte r 2.3.1) and therewith are neither persistent, nor do they bioaccumulate or induce toxic effects outside of the direct treatment chamber. Of potential concern are Other Substances, which can theoretically be indirectly produced by the applied method as reaction products within the pro cess. Whether the statistic results of measured Nitrite values showed, the treat ment process is not responsible for detected Nitrite values,

however the risk characterization will focus on Nitrite.

5.1.1 Persistence (G9: 5.1.1.1)

Nitrite is a nutrient and either nitrified or denitrified quick ly ñ depending on whether aerobic or anaerobic conditions prevail. A half-lif e of 10 minutes has been estimated.

5.1.2 Bioaccumulation (G9: 5.1.1.2)

Bioaccumulation of Nitrite by freshwater fish is higher than by marine fishes because Nitrite competes with chloride for uptake channel proces ses. No bioaccumulation factor is available.

5.1.3 Toxicity (G9: 5.1.2.3)

In general, few studies of chroni c toxicity are considered to be reliable, and often documentation of experimental set-ups is poor.

Information on acute toxicity is more abundant.

Toxicity of Nitrite to fish even occurs at natu ral concentrations or when nitrification becomes unbalanced due to anthropogenic ac tivities. The toxicity of Nitrite depends on a large number of factors, the most important being water chemistry and especially chloride concentration. Data for acute toxicity are shown in Table 2.

5.1.4 PBT Criteria (G9: 6.4.1, Table 1)

In Table 5 the PBT criteria (Persistence, Bioacc umulation and Toxicity) are shown for Nitrite. Compared with data from Procedure (G9), Table 1: Criteria for identification of PBT substances these results show that persistence and bioaccumu lation rates for the selected substances do not exceed the criteria from Table 1 in Procedure (G9).MEPC 59/2/8

ANNEX Page 30

I:\MEPC\59\2-8.doc Other substance Persistence Bioaccumulation Toxicity

Nitrite Turnover has been approximated to be less than a day to more than 10 days (latter in oligotrophic environments). No BCF has been

determined Data for acute toxicity

are shown in table 2

Table 5: Summary of persistence, bioaccu mulation and toxicity criteria for Nitrite

5.2 Evaluation of the treated ballast water (G9: 5.2)

To evaluate the impact of ballast water treatm ent on the production of Other Substances samples were analysed chemically before and after the treatment process.

Test medium Salinity (PSU)

Freshwater 0

Brackish Water 18.9 ñ 19.5

Saltwater 31.6 ñ 33.1

Table 6: Range of salinities during sampling inve stigating by-products with temperature ranges
from 16.1 to 17.8°C.

Treatments were carried out at three different salinities (Table 6) and with two to three parallels in several runs (two to four) each before (pre) and after (post) treatment. All TRO, HAA, THM, LHKW, Acetonitrils and Bromophe nols measured in samples before and after treatment were below Levels of Quantification.

Except for Chloroform, where some results for samples before and after treatment at or slightly above the LoQ (0.02 μg/l) were measured. These values indicate, that Chloroform background

concentrations within the test water in the Bremerhaven harbour waters are to characterize around 0.02 μg/l. From this results can not be conc luded, that the treatmen t process cause higher

levels for Chloroform. For this reason values near to the LoQ will not be considered in further discussion. An exemption is the detection of one value for Chloroform concentration before the treatment. As this is the only higher value clea rly above LoQ and was measured in the sample before the treatment, it is considered as an outlier without further concern for the evaluation.

AOX could be measured in some samples at very low concentrations at the LoQ (0.02 mg/l), in samples before and after treatment and here th e same can be applied as for Chloroform.

Nitrite concentrations especially for brackish and salt water ar e above the LoQ and show a high variability in samples in both treatments, before and after the BWTS. Rega rding this variability and to obtain a significant conclusion, if the data from samples before treatment are different compared to data after treatment a statistic analysis has been performed. For the statistic evaluation the program SYSTAT 12 was used. Data of freshwater samples could not be tested, because the variability was too low, to apply test statistic for investigation of significance of the variability. Because the data of brackish and salt water samples were not normally distributed, the normally non-parametric Mann-Whitney U Test Statistic incl uding the appropriate descriptive statistic was applied.MEPC 59/2/8

ANNEX Page 31

I:\MEPC\59\2-8.doc Figure 3: Box plot of descrip tive statistic as part of the Mann-Whitney U Test Statistics of

Nitrite concentrations before and after treatme nt of brackish water (Vor: before treatment, Nach: after treatment, middle line in the box: median, outer lines of the box plot: 25% and 75% quartile ñ re presenting 25% or 75% of the data within this range, n=14)

Figure 4: Box plot of descrip tive statistic as part of the Mann-Whitney U Test Statistics of

Nitrite concentrations before and after treat ment of salt water (Vor: before treatment, Nach: after treatment, middle line in the box: median, outer lines of the box plot: 25% and 75% quartile ñ re presenting 25% or 75% of the data within this range, n=14)

In Figures 3 and 4 the box plots (descriptive statistic) of the estimation are shown. Number of

data for brackish as well as for salt water for calculations before and after treatment were 14.

The result of the test statistic shows the estima ted P-values (0.27 for brackish water, 0.16 for salt water) were below the specified significance level of 95% ( α = 0.05). It can be concluded that Nitrite concentration from samples before and after the treatment belong to the same data population ñ means they are significantly not differe nt. From this it can be concluded that the Nitrite concentrations measured in samples were not generated by the treatment process.

All Nitrite values before and after treatmen t are still within the German drinking water regulation, where 0.5 mg/l are the maximal allowed c oncentration at the use rís tap. Hence no risk

to human health and/or to the marine environm ent can be deduced on the basis of these data.MEPC 59/2/8

ANNEX Page 32

I:\MEPC\59\2-8.doc 5.3 Risk characterization and analysis (G9: 5.3)

5.3.1 Reaction with organic matter (G9: 5.3.3)

Nitrite does not react with organic matter.

5.3.2 Characterization of degrad ation route and rate (G9: 5.3.5)

Half-life of Nitrite has been estimated to be around 10 minutes, but tu rnover times strongly depend on the ambient conditions, esp ecially the activity of nitrifying or denitr ifying bacteria and the oxygenation of the water. As Nitrite is an essen tial part of the ni trogen cycle, their consumption is fast.

5.3.3 Prediction of discharge and envi ronmental concentrations (G9: 5.3.8)

On the basis of the chemical data determined in samples from the treated ballast water there is no

indication of release of harmfu l substances, resulting in an increase of environmental

concentrations.

5.3.4 Assessment of potential fo r bioaccumulation (G9: 5.3.7)

As there is no indication for the release of harmful substances, the actual potential for bioaccumulation is considered to be very low.

5.3.5 Effect assessment on aqua tic organisms and sediment

Due to their extremely low lifetime, direct effects of photons and cavitation bubbles are not anticipated. In the effect assessment, only Ot her Chemicals as Nitrite are considered.

5.3.5.1 Risks to aquatic organisms

Acute and chronic toxicity data ha ve been displayed in chapter 2.3.2. The toxicity of Nitrit e depends on a large numbe r of factors, the most important being water chemistry and especially chloride concentratio n. With higher chloride concentration, toxicity decreases. Accordingly Schwˆrbel (1991) pr oposed the following envi ronmental criteria:

0-3 mg Cl - /l ! 33 μg/l NO 2 - <3 ñ 7.5 mg Cl - /l ! 164 μg/l NO 2 - <7.5 - 15 mg Cl - /l ! 296 μg/l NO 2 - <15 ñ 30 mg Cl - /l ! 394 μg/l NO 2 - >30 mg Cl - /l ! 493 μg/l NO 2 -

Bursch (2003) suggested a PNEC of 0.01 mg/l. This value corres pond with the guidance values

of the Fish Directive of the EU for Salmonid and for Cyprinid waters of ≤ 0.01 mg/l NO 2 - and ≤ 0.03 mg/l NO 2 -, respectively (EU Directive 2006/44/EC).MEPC 59/2/8

ANNEX Page 33

I:\MEPC\59\2-8.doc 5.3.5.2 Risks to sediments and for bioaccu mulation and secondary poisoning

Since Nitrite as an ion and therewith completely water soluble, for Nitrite does not exist a log

K OW value. Therefore, neither sediment toxicity , nor bioconcentration or magnification effects, including secondary poisoning are c onsidered to be of concern.

5.3.5.3 Risks to humans

No effects on humans are anticipated during th e process (see Human health risk chapter). Exposure to treatment water could occur accidentally (e.g., people falling overboard). As described above, concentrations of Other Chem icals in the treated balla st water are, however,

considered to be too low to cause effects in aquatic organisms and will be unlikely to cause effects to humans that may become accidentally e xposed to a mixture of treated ballast water and

water.

Nitrite is strictly regulate d in drinking water due to its known interference with the oxygen-carrying capacity of the bl ood which poses especially small children in danger.

Drinking water standards from the WHO, EP A, EU and Germany are depicted below (partly additional conditions with regard to Nitr ate are included, which have been omitted here, as the focus is on Nitrite).

Regulatory body Nitrite

WHO 3 mg/l 3 Guideline value

EPA 1 mg/l 4 Maximum contaminant level

EU 0.5 mg/l 5 Quality standards have to be set by the member

states but should not exceed the values of Appendix A of the drinking water directive

Germany 0.5 mg/l 6 The water that leaves th e water treatment facility

must not exceed Nitrite concentrations of 0.1 mg/l

Table 7: Drinking water standards from th e WHO, EPA, EU and Germany for Nitrite

Derivation of guidelines for Ni trite contamination of drinki ng water are usually based on calculation of uptak e per day per kg.

Extract from the WHO Dr inking water guideline:

The guideline for Nitrite of 3 mg/l as Nitrite is based on human dat a showing that doses of Nitrite that cause methaemoglobinaemia in infants range from 0.4 to more than 200 mg/kg of body weight. By applying the lowest le vel of the range (0.4 mg/kg of body weight), a body weight of 5 kg for an infant and a drinking-water cons umption of 0.75 litre, a guideline value of 3 mg/l (rounded figure) can be de rived (WHO/SDE/WSH/07.01/16).

3 WHO/SDE/WSH/07.01/16

4 http://www.epa.gov/OGWDW/contaminants/dw_contamfs/nitrates.html

5 COUNCIL DIRECT IVE 98/83/EC

6 Trinkwasserverordnung 2001, chemische Parameter IIMEPC 59/2/8

ANNEX Page 34

I:\MEPC\59\2-8.doc 6 RISK ASSESSMENT

As per definition ìriskî is determined by the equation

Risk = Probability of occurrence * extent of loss/impact

In the following the potential risks cause d by the operation of the AquaTriComb  ballast water treatment system technology as to the safety of the ship and to human health will be assessed.

This will be carried out by means of estimati ng the probability of o ccurrence of hazardous situations multiplied with the estimations on the extent of losses and/or scale of impact caused by

such hazardous scenarios.

By definition of ìprototypingî this risk assessment has to be a probabilistic approach rather than a deterministically based risk assessment. This, in turn, may result in the absence of data-base secured figures. In case of such lack of data-based figures a prescriptive approach will be applied.

6.1 Risk to safety of ship

6.1.1 Ship integrity and the BWTS

The AquaTriComb  plant will be installed in line after the shipís ballast water pump(s). Inlet and outlet piping are equipped with by-pass-valves and shut-off sa fety valves (Figure 1). The valves are fitted with flanges and will be equipped with electrical ly driven, and manual actuators.

If, for certain reasons, the BWTS operations have to be interrupted (service, maintenance, etc.) the plant will be bypassed by means of the bypa ss-valves and closing of the safety valves. This design does not interfere with the shipís integrity.

Although electrical load calcu lations on larger versions of the pl ant have not been finalized yet, the probability that the energy demand would in terfere with the onboard power supply, that is, the auxiliary diesel-generator sets, is rather low.

With regard to the additional weight the prototyp e version is at approxima tely one point six (1.6)

tons at 250 m3/h volume flow rate. A slightly flattened linear increase is expected for the larger versions, for example the 2,000 m3/h version would be at approximately 10 to 12 tons. This weight equals half the gross weight of a lo aded twenty-foot standard container (20' container maximal gross weight equals 24 tons). Therefore, also the additional weight of the BWTS would not impact the integrity of the ship.

6.1.2 Corrosion

The AquaTriComb  technology is characterized by the absence of any Preparations or aggressive additives or generated substances. Therefore, no corrosion supporting, or corrosion stimulating effects are c onnected to the technology.

The effects of acoustic cavitation, which are generated by the ultr asonic oscillation, are strictly

restricted to the interior of th e MicroSintFilter (MSF) modules, re spectively. Transfer of acoustic cavitation is system-dependent excluded, and pi ping or other components of the ship are not

effected.MEPC 59/2/8

ANNEX Page 35

I:\MEPC\59\2-8.doc All the internal compone nts of the AquaTriComb  BWTS are designed a nd construed to match

the effects of acoustic cavitation.

6.1.3 Fire and explosion

The electrical and electroni c parts of the AquaTriComb  BWTS are intrinsically safe. There is no risk of fire or explosion associated with the system.

6.1.4 Noise and vibration

The noise level generated by the AquaTriComb  BWTS is far below the permissible average noise level at the work pla ce of 85 decibel (A), accordi ng to EC guidelines 2002/44/EG and 2003/10/EG (BGBl 2007).

Vibration level caused by the ultras onic oscillation is rather low and noticeable only in direct

contact with the MSF modules a nd the UV-reactor (hand touch).

In addition, the plant wi ll be installed on vibration mounts fo r the protection of the plant itself

and to reduce in-water noise emissions to a minimum.

6.1.5 Conclusion: risks to safety of ship

The risks to the safety of the ship, caused by the installation and operation of the BWTS AquaTriComb  , which is defined by the probability of the occurrence of a dangerous situation originated by the plant and the extent of imp act, is not measurable by means of non existence.

This covers the shipís integrity, corrosion, fire and explosion as well as noise and vibrations. If the ship itself makes for a dangerous scenario like fire, explosion, struct ural damage or loss, etc., the risks originated by the BWTS remain at zero as intrinsic safety is maintained and power shut-off is most probable.

6.2 Risks to human health

The AquaTriComb  technology exclusively applies non-ha zardous physical means of ballast water treatment which pose no ri sks to human health, that is:

- Ultraviolet radiation (UV); and - Ultrasonic oscillation (US).

The application of UV and US, however, generate s physical active processe s, explicitly photons and cavitation bubbles, which in turn could resu lt in OH radicals (Relevant Chemical) and by-products (Other Chemical).

The generation of OH radicals and by-products ar e confined to the feasible minimum by means of the application of exclusiv ely UV-C radiation at 254 nm wave length. Also, by the application of US-oscillation at a low frequency for a very limited time period of less than one (1) second only the size of the cavitation bubbles is as sm all as possible, and the effect of OH radical generating physical sonoluminescence is re duced to the most practicable minimum.MEPC 59/2/8

ANNEX Page 36

I:\MEPC\59\2-8.doc Therefore, OH is considered not resident in the treated ballast water and causes no harm to

human health or to the environment.

In case OH radical related eco-toxi cological by-products (Other Ch emicals) are generated this will be on a level below the LoQ of the analytical methods. Thus by-products were not detectable in ballast water treated by A quaTriCombô BWTS. In addition, o ccurrence of UV radiation and Ultrasound are limited to the hermetically encl osed MSF and UV-reactor, and risks to human

health can be excluded.

6.2.1 Human exposure assessment

Because exclusively physical means of ball ast water treatment are applied with the AquaTriComb  technology the exposure assessment focuses on potential exposure as to:

- UV-radiation; - Ultrasound oscillation; and - OH and by-products.

Potential exposure to Ultraviolet (UV) radiation : Inside the hermetically enclosed UV-reactor UV-C-radiation is generated by high performance, low-pressure mercury germicidal lamps with a filtered and selected wavelength of approximately 253.7 nm.

The tube casing of the AquaTriComb  UV-reactor and the top and bottom plating is made of stainless steel (V4A). Top plating cable fair leads are welded and additionally covered by aluminium coverage. All welding and bolt conn ection are according to respective technical norms and standards and performed by accredited and certified workmanship. Therefore, the probability of regular and/or accidental emission of UV-radiation equates zero.

Potential exposure to Ultrasound (US) oscillation : The ultrasonic tube oscillators are located along the centre line inside th e MicroSintFilter (M SF) modules and insi de the UV-reactor. Oscillation at low frequency permanently preven ting particle accumulation in the MSF modules, and des-agglomeration of orga nic particles, detr itus, colony-forming algae and other agglomerates (bio film) at the surfac e of the mercury germicidal lamps.

The MSF modules are identical in design and constr uction as the UV-reactor, which results in no release of ultrasound and no impact to human health.

Potential exposure to OH and by-products : As stated above, the concentration of OH and by-products would be on such a lo w level that risks to human heal th can be excluded. Therefore, risks to human health by mean s of exposure to Relevant or Other Chemicals can be excluded.

In applying the Margin of Safety (MOS) or Ma rgin of Exposure (MOE) approach, the equation would consequently result in No Obse rved Adverse Effect Level (NOAEL).MEPC 59/2/8

ANNEX Page 37

I:\MEPC\59\2-8.doc 6.2.1.1 Hazard identification

In regular operation mode of the AquaTriComb  BWTS no hazards to human health can be

identified:

- No persons are exposed to UV radiation;

- Low-level casing vibration caused by US oscillation inside MSF modules does not pose any hazard; and

- No contact with non-hazardous concentr ation of potential Relevant or Other

Chemicals.

In service and maintenance (S&M) mode hazards are limited to illegal and/or intentional risk

exposure:

- Non switch-off of energy supply before S& M works with the risk to exposure to UV-C radiation; and

- Operational procedures, alert signing and emergency shut-off device will prevent such scenario (to be integrated into shipboard SMS-manual 7 ).

In incidental/accidental mode such as shipboard fire, explosion, black-out, structural loss, etc. most probably energy supply to BWTS will be cut-off anyhow and/or BWTS will have

malfunction.

6.2.1.2 Exposure assessment

Regular operation and maintenance procedures (O &M) on board ship include inspection of sea chest and sea water/ballast water piping and pump s, and related services. For such action the BWTS will be shut-down, thus no exposure to UV-ra diation or chemicals will appear. This also counts for dry docking and/or annual classificat ion services in dock/ alongside the pier.

Due to high quality plant components and materials service and maintenance intervals of the BWTS are extremely low. The exchange of the u ltrasonic tube oscillators are calculated after a minimum of approximately 16,000 operational hours, the exchange of the low-pressure mercury germicidal lamps are calculated af ter circa 12,000 operational hours.

As no Preparations are utilized or generated by the AquaTriComb  BWTS normal and good workmanship practice is to be applied by th e personnel involved (gloves and mask).

No additional or special precaution meas ures are recommended or necessary.

6.2.2 Conclusion: Potential Human Health Risks

Normal operation and maintenance (O&M) procedures of the AquaTriComb  BWTS do not pose any risks to human health, nor acute, ne ither mid/long-term exposure situations are realistically probable. This statement covers all population groups including crew, service

7 SMS means Safety Management System according to the ISM-Code (ISM 2002).MEPC 59/2/8

ANNEX Page 38

I:\MEPC\59\2-8.doc technicians, passengers and pe rsonnel, public and port persons , or consumers of seafood and persons at the coastline.

The probability of incidental and/or accident al health risks caused by the AquaTriComb  BWTS are limited to catastrophic or worst-case scenarios, although almost implausible.

The absence of any Preparations reduces such ri sks to shipboard fire, explosion, structural damage/loss and similar casualties. In occurrence of such catastrophic events it is assumed that the power supply to the BWTS is cut-off anywa y, resulting in non-perfor mance of UV-radiation and/or US-oscillation, and associated processes.

From the Risk Assessment carried out are no id entifiable risks associated to the ship, its personnel or other persons and/or parties concer ned, which are originated by the installation and operation of the AquaTriComb  ballast water treatment system.

This is mainly due to the fact that the BWTS does not introduce any Preparations, Active

Substances or Relevant Chemicals but the tech nology is a purely physical treatment technology. Further, the design and construc tion do not interfere with the vesselís integrity, and high-end components and materials guarantee reliable and sound performance.

6.3 Risks to the aquatic environment

6.3.1 Magnitude of Hazard from the Ballast water treatment process

In Table 8 is exemplary shown the PNEC (Predict ed No Effect Concentrat ions) for Nitrite value

in water.

Substance Substance class PNEC Medium Reference

Nitrite Inorganic nutrient 10 μg/l Water Bursch 2003

Table 8: Predicted No Effect Concentr ations (PNEC) for Nitrite (Bursch 2003).

6.3.2 Probability of exposure

Exposure to Other Chemicals could occur through the emitted treated ballast water. According to the chemical analyses, none of the Other Chem icals, discussed in this report, are produced significantly above LoQ. It can al so be concluded from the chemical data directly that the Nitrite values shown by statistic evaluation ar e not caused by the treatment process.

6.3.3 Assessment of environmental risk

On the basis of the results of analysis of severa l theoretical possi ble by-products at this stage of

the assessment it can be concluded that there is no indication for an environmental risk from this

treatment process.

6.3.4 Conclusion on risk to human health

Exposure of humans to the active substances is not possible. Exposure to potentially Other

Substances in the treated ballast water would th eoretically be possible but with low probability. Effects are highly unlikely due to the low aquatic toxicity.MEPC 59/2/8

ANNEX Page 39

I:\MEPC\59\2-8.doc Toxic effects through volatile subs tances such as chloroform woul d theoretically be possible and

there is indication of risk to human health from any substances produced in the process at this

stage.

7 ASSESSMENT REPORT (G9: 4.3)

7.1 Overview of data and endpoints

During the AquaTriComb  -process aquatic organisms will be affected by photons and cavitation bubbles. Both are short lived and their presence is restricted to the duration of the UV and US application, locally to the treatment reactor.

Exposure to the aquatic environment from substa nces termed Other Chemicals, which can be produced indirectly through photons or cavitation bubbles, could occur after discharge of the

treated ballast water.

As shown by chemical analysis of water sample s before and after ballast water treatment,

concentrations of all observed substances we re below the LoQ. Ther efore, no indication of release of harmful substances, resulting in an in crease of environmental concentrations could be observed. Except for Nitrite, which data above the LoQ showed a hi gh variability of concentrations. Statistical analysis indicate that data before and after treatment are not significantly different. Therefore, the treatment process is not res ponsible for the observed Nitrite

values.

As there is no indication for the release of harmful substances, the actual potential for bioaccumulation is considered to be very low. Ne ither sediment toxicity, nor bioconcentration or magnification effects, including secondary poiso ning are considered to be of concern.

From the Risk Assessment carried out are no id entifiable risks associated to the ship, its personnel or other persons and/or parties concer ned, which are originated by the installation and operation of the AquaTriComb  ballast water treatment system.

On the basis of the results of analysis of se veral theoretical possible by-products and the risk assessment it can be concluded that there is no indication for a risk to the aquatic environment and/or humans based on the treatment process of the AquaTriComb  BWTS.

7.2 Test report data quality

References used for this approval comprise Ri sk Assessment studies from Euro Chlor, the European Commission, WHO, and US EPA, which critically evaluated th eir data basis with regard to accredition of laboratories, standard ization of test systems and documentation of

results.

Furthermore, peer reviewed articles were used to gain detailed information. The PAN Pesticide database proved to be very helpful. All of it s information is backed up by rigorous scientific

studies and most of the data are taken from o fficial sources of weight-of-the-evidence-type evaluations when they are available. For more information of data accuracy of the data base: http://www.pesticideinfo.org/Do cs/data.html#AccuracyofDataMEPC 59/2/8

ANNEX Page 40

I:\MEPC\59\2-8.doc 8 REFERENCE DOCUMENTS

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