I:\MEPC\57\2-7.doc For reasons of economy, this document is pr inted in a limited number. Delegates are kindly asked to bring their copies to mee tings and not to request additional copies. INTERNATIONAL MARITIME ORGANIZATION

IMO 

# E

MARINE ENVIRONMENT PROTECTION

COMMITTEE
57th session Agenda item 2 MEPC 57/2/7

20 December 2007 Original: ENGLISH

HARMFUL AQUATIC ORGANISMS IN BALLAST WATER

Application for Basic Approval of a combined ba llast water management system consisting of

sediment removal and an electrolytic process using seawater to produce Active Substances (Greenship Ltd)

Submitted by the Netherlands

SUMMARY

Executive summary: This document contains non-confidential information for Basic Approval of a combined system consisting of sediment removal and an electrolytic process using seawat er to produce Active Substances, as required by the Procedure for approval of ballast water management systems that make use of Active Substances (G9). Relevant documents together with the application dossier will be

submitted to the Marine Environmen t Division of the Organization for review by the GESAMP-Ballast Wa ter Working Group in accordance with Procedure (G9).

Action to be taken: Paragraph 6

Related documents: BWM/CONF/36; MEPC 53/24/Add.1; MEPC 55/2/16; MEPC 56/2/2

and BWM.2/Circ.10

Introduction

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

2 The Procedure for approval of ballast wate r management systems that make use of Active Substances (G9), as adop ted by resolution MEPC.126(53), defi nes the principal aspects to be documented by data or tes ting (MEPC 53/24/Add.1, annex 4, pa ragraph 4.2.1) and some basic principles for risk evaluation (MEPC 53/24/ Add.1, annex 4, paragraph 5.3). According to section 6 of Procedure (G9) th e Organization should evaluate th e information provided in the application.MEPC 57/2/7 - 2 -

I:\MEPC\57\2-7.doc 3 According to circular BWM.2/Circ.10, pr oposals for approval of ballast water management systems that make use of Active S ubstances are to be submitted to the Marine

Environment Division of the Or ganization to be reviewed by the GESAMP-Ballast Water Working Group. In the meantime, a document on the proposal for approval containing all the

non-confidential data related to the respective ballast water management system that makes use

of Active Substances needs to be submitted to the MEPC.

4 The competent authority in the Netherlands ha s verified the applicat ion dossier prepared by Greenship Ltd. and believes it to satisfy the data requirements of Procedure (G9).

5 The Netherlands therefore submits the non-confid ential part of the applicantís dossier in the annex to this document to the Organization for evaluation according to Procedure (G9). The complete dossier will be made available to the experts of the GESAMP-Ballast Water Working Group.

Action requested of the Committee

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

***MEPC 57/2/7

I:\MEPC\57\2-7.doc ANNEX

NON-CONFIDENTIAL INFORMATION ON THE COMBINED BALLAST WATER

MANAGEMENT SYSTEM CONSISTING OF SEDIMENT REMOVAL AND AN

ELECTROLYTIC PROCESS USING SE AWATER TO PRODUCE ACTIVE

SUBSTANCES DEVELOPED BY GREEN SHIP LTD (THE NETHERLANDS)

This annex follows the requirements for app lication as presented in the Methodology for information gathering and the conduct of work of GESAMP-BWWG (the Methodology, MEPC 56/2/2, annex 4) and has the following content:

Chapter 1: Introduction

Chapter 2: Description of the system and th e production of Active Su bstance and Relevant

Chemicals

Chapter 3: Data sets and risk characterization of Active Substance and Relevant Chemicals (Methodology, sections 3 and 6)

Chapter 4: Toxicity evaluation of treated ballast water

Chapter 5: Risk assessment (Methodology, section 7)

Chapter 6: Assessment report (Methodology, section 8)

References

NB: The applicant has not reported on sect ions 4 and 5 of the Methodology (MEPC 56/2/2, annex 4). Since no chemical s are added a material data safety sheet seems redundant. The information requirements as described in chap ter 4 are either redundant or will be provided in the application for Final Approval (e.g., technical manual).

1 INTRODUCTION

The applicant proposes a ballast water management system (BWMS) that combines removal of sediment at ballasting with disi nfection, using seawater in an el ectrolytic process to produce

Active Substances for the deactivation of biot a, at ballasting and de-balla sting. This ap plication is based on the Procedure for approv al of ballast water management systems that make use of Active Substances (G9).

The BWMS is capable of removing sediment-particles > 20 μm w ith an efficiency of 100% and sediment-particles of > 10 μm with an efficiency of > 80%. The treated water is subsequently disinfected by Active Substances generated in an electrolytic cell with a killing efficiency of over 99.99%. Upon completion of treatment, the balla st water complies with the D-2 standard of the BWM Convention. The capacity of the BWMS is modular: it can handle ballast water flows ranging from 50 m 3 /hr up to 5.000 m 3 /hr and even more.

The seawater electrolytic process produces Free Active Chlorine (FAC) from natural seawater taken on board. The applicantís in-line disinfecting technique can be successfully implemented in a wide range of applications. Using the salt present in the water, the electrolytic cell produces 0.5 ñ 3 ppm FAC. FAC is widely applied for the disi nfection of water. The majorMEPC 57/2/7

ANNEX Page 2

I:\MEPC\57\2-7.doc advantage of this type of elect rolytic cell is the long- lasting coating. The el ectrolytic cells can disinfect with a minimum of maintenance (a once a year cleaning is all that is required) for the duration of approximately three to five years.

The applicant believes that the BWMS is a sa fe and economical system for ships and crews applicable without the storage of chemicals on board. Water chlori nation is a safe and reliable disinfection procedure used worldw ide for drinking water production.

After FAC has deactivated micro-organisms in ba llast water, it will decompose rapidly into harmless chloride ions, a natural component of seawater, through spontaneous decomposition by reactions with organic matter in natural seawater . FAC has no persistence or bioaccumulation in

water.

The applicant evaluated the toxicity of seawater treated with electrolytically generated FAC by toxicity tests and using literature data. The results of these evaluations indicate that the influence

on the environment of the di scharges after treatment by the BWMS is negligible.

2 DESCRIPTION OF THE SYSTEM AND THE PRODUCTION OF ACTIVE

SUBSTANCE AND R ELEVANT CHEMICALS

Greenshipís Ballast Water Management System (BWMS)

Greenshipís BWMS consists of a multi-hydrocyclone system for removal of larger particles and organisms, followed by disinfection with an electrolytic cell. Th e electrolytic cell produces FAC from chloride in seawater, which is extremel y potent in breaking down micro-organisms and bacteria. No substances are added to the process.

2.1 General description of the system

2.1.1 System Components

The BWMS consists of the follo wing two modular components:

- the Sedimentor: The sediment removal system removes sediment and biota during intake; resulting in 100% removal for particles of 20 micron and bigger
and 80% removal fo r particles of ≥ 10 micron. The remaining sediment particles stay in suspension and will leave the ba llast tanks during de -ballasting; and

- the Termanox : The electrolytic cell decimate s all bacteria and organisms. Together with the Sedimentor the electrolytic cell achieves a killing rate
of 99.99%.MEPC 57/2/7

ANNEX

Page 3

I:\MEPC\57\2-7.doc Figure 2.1: Picture of the 50m 3 /hr laboratory-based test unit for ballast water treatment with a Sedimentor and an Electrolytic cell

The sediment removal system ìSedimentorî

Greenship developed a revolutionary system pr eventing sedimentation of suspended solids in ballast tanks: not only does the system fight the pollution of the ballast wate r; it even prevents the cause of the consolidating mud problem. With tons of sediment in the ballast tanks, a ship consumes more fuel and transports less cargo. In r ecent calculations, it appears that there is an economic profit on fuel consumption up to 3% a nd even more through separation of sediment from ballast water (source: Chemgas, the Nether lands) which provides a very attractive pay back.

2.1.2 The electrolyse system: Termanox

The electrolytic cell kills effec tively bacteria and organisms. T ogether with the Sedimentor, the electrolytic cell achieves a killi ng rate of 99.99%. This secondary stage of the BWMS consists of a dedicated electrolytic cell, which was especial ly engineered for this specific application. The ìkilling efficiencyî of this el ectrolytic process is based on the electrolysis of chloride from seawater. Greenshipís disinfecting technique can be successfully im plemented in a wide range of applications. Using the chloride in the wate r, the electrolytic ce ll produces hypochlorite. The Termanox is like the Sedimentor engineered from modular components. The system enables the construction of any capacity required in m 3 /hour, and can be tailored to the same volume requirements as the Sedimentor.MEPC 57/2/7

ANNEX Page 4

I:\MEPC\57\2-7.doc For the optimum performance of the electrolytic cell, it is of great importance to reduce the amount and size of sediment particles. Fouled seawater may lead to deposits build ing up on the

anodic and cathodic electrodes effec tively reducing their efficiency. By using the Sedimentor in combination with the electrolytic cell, a trouble-free operation can be guaranteed.

The Electrolysis of Aqueous NaCl

Figure 2.2: An idealized drawing of a cell in which an aqueous solution of sodium chloride is electrolysed

NaCl (s) + H2O (l) ! Na + (aq)+ Cl - (aq)

The Na + ions migrate toward the negative electrode and the Cl - ions migrate toward the positive

electrode. There are two substances that can be reduced at the cathode: Na + ions and water

molecules.

Cathode (-):

Na + + e - Na E o red = -2.71 V

2 H 2 O + 2 e - H 2 + 2 OH - E o red = -0.83 V

Because it is much easier to reduce water than Na + ions, the only product formed at the cathode is hydrogen gas.

Cathode (-): 2 H 2 O( l ) + 2 e - H 2 ( g ) + 2 OH - ( aq )

There are also two substances that can be oxidized at the anode: Cl - ions and water molecules.

Anode (+):

2 Cl - Cl 2 + 2 e - E o

ox = -1.36 V

2 H 2 O O 2 + 4 H + + 4 e - E o

ox = -1.23 V

The standard-state potentials for these half-reacti ons are so close to each other that we might expect to see a mixture of Cl 2 and O 2 gas collect at the anode. In pr actice, the only product of this

reaction is Cl 2

. Anode (+): 2 Cl - Cl 2 + 2 e In summary, electrolysis of aqueous NaCl solutions gives a mixture of hydrogen and chlorine gas and an aqueous sodium hydroxide solution.

2 NaCl( aq ) + 2 H 2 O( l ) 2 Na + ( aq ) + 2 OH - ( aq ) + H 2 ( g ) + Cl 2 ( g )MEPC 57/2/7

ANNEX

Page 5

I:\MEPC\57\2-7.doc The products of the electrolysis of aqueous sodium chloride react with water to form sodium hypo-chlorite.

Cl 2 ( g ) + H2O( l ) HOCl( aq ) + H + (aq) + Cl - ( aq )

HOCl (aq) H + (aq) + ClO - (aq)

2.2 Results of laboratory tests

2.2.1 Test unit

For determining the performance of the BWTS, a te st was executed with a laboratory test unit with a capacity of 50 m 3 /h. The experiments took place at the NHL University for Professional Education, Leeuwarden, The Netherlands on 22 June 2005 and were witnessed by Mr. T. Henstra of Lloydís Register EMEA.

The feed was technical seawater which consiste d of a suspension of sediment collected from ballast water tanks from several ships. The f eed is separated with the Sedimentor in a concentrated underflow with the bigger suspended so lids which is returned to the sea (in the test this is a buffer tank) and an overflow with the smaller suspended solids which was treated with the electrolytic cell for disinfection of the water a nd transported to the ballast tank/sea (in the test this is a buffer tank or sewer).

2.2.2 Purpose of the experiments

The test is divided in two experiments, namely:

Sedimentor test: In this test the separating efficiency of the Sedimentor is determined with the following conditions:

- pressure difference of 2 bar and a reflux of 0,3 (the fraction which is returned to

the sea = underflow divide d by the sum of the overflow)

- sediment suspended in feed in a concentration between 1-5 wt%

- salt concentration of 35 PSU

Killing test: In this test the killing efficien cy of the electrolytic cell is determined on heterotrophic bacteria in a minimum con centration of 104 colony forming units per millilitre (cfu/ml). The conditions of the Sedime ntor and the technical seawater are equal

to the conditions in the Sedi mentor test. The electrolytic cell is used with a maximum current output (=25 Ampere).

2.2.3 Results and discussion

Based on the results of the Sedimentor test, it can be concluded that 100% removal is achieved for particles of 20 μ m and bigger. For particles of 10 μ m, 80% removal is achieved. This means that the particles that pass the el ectrolytic cell vary from 0 ñ 20 μ m. But the biggest load is

smaller than 10 μ m, namely, 80%. Viable organisms incl ude thereby mostly plankton, algae, bacteria and viruses. These organisms can be e ffectively treated with the electrolytic cell (see table 2.1).MEPC 57/2/7

ANNEX Page 6

I:\MEPC\57\2-7.doc Figure 2.3: Separation curve for the Sedimentor

According to the Guidelines for approval of ballast water management systems (G8), test organisms may be either naturally occurring in the test water, or cultured species that may be added to the test water (paragraph 2.3.18 of Guid elines (G8)). For influent water, heterotr ophic bacteria should be present in a density of at least 10 4 living bacteria per millilitre (paragraph 2.3.19.3 of Guidelines (G8)). Samp les during the test should be taken at the following times and locations: immediately be fore the treatment equipment, immediately after the treatment equipment and upon discha rge (paragraph 2.3.25 of Guidelines (G8)).

In order to be sure to obta in a final concentration of 10 4 cfu/ml indicator bacteria in the buffer

tank of the test unit, 300 ml of E. coli in a concentration of 10 8 cfu/ml has been added. After determining the total amount of bacteria in the buffer tank the concentration tu rned out to

be 10 5 cfu/ml. Sampling took place simultaneously be fore and after the el ectrolytic at three points in time, namely, after 1, 2 and 3 minutes . Samples were immediat ely processed in the laboratory.

Results of this test of the electrolytic cell unit show a killing effici ency of 99.99% (or log 4).

The influent test water contained a re presentative amount of bacteria (10 5 cfu/ml) and at a setting of 25 Ampere, the colony count, after passi ng the electrolytic unit, showed after 1 minute 30 colonies and after 2 and 3 minutes less than 10 colonies per milliliter (see table 2.1). According to regulation D-2 of the Convention, Vibrio cholerea should be less than 1 cfu per 100 ml ballast water, whereas E.coli must be less than 250 cfu/ml and intestinal

Enterococci must be less than 100 cfu/ml. Separation curve

0 20 40 60 80 100 120

0 20 40 60 80 100 diameter (μm)percentage ∆ P = 2bar; reflux = 0,3;

salt content = 35 PSU; Temperature
= freeMEPC 57/2/7

ANNEX

Page 7

I:\MEPC\57\2-7.doc Table 2.1: Killing efficiency of electrolytic cell

Concentration sodium hypochlorite produced with 25 A[ppm] Colony Count [CFU/ml]

10,8 Before electrolytic cell After electrolytic cell Killing efficiency (%)

sample 1

sample 2

sample 3 1,80E+05 30

2,10E+05 < 10

2,60E+05 <10 99.983

99.995

99.996

Medium control

Blank No growth

1,20E+05 conclusion: log 4-5 (99.99%) killing efficiency

Legend Medium control: Non-inoculated plates

Blank: Total count on sample from buffer tank before addition of test organism

From this result it can be concluded that the killi ng efficiency of the electrolytic cell is sufficient for the IMO regulated organisms smaller than 10 μ m. Minimum load to be tested is, namely, 10 4 cfu/ml. With a log 4 killing efficiency and a load of 10 4 cfu/ml, the concentration bacteria after the electrolytic ce ll would theoretically be 1 cfu pe r 100 ml. In practice the average

will be less than 1 cfu/ml so it meets also the standard for Vibrio cholerea . After all lab-tests

showed that Vibrio cholerea is more sensitive for hypochlorite than other bacteria.

2.2.4 Final remarks

Sediment layers that precipitate in ballast tanks fo rm a substrate suitable fo r the survival of many benthic species and dormant stages of plank toni c species, while they decrease the effective volume of ballast tanks. The results prove that chlorination treatment is effective in killing

bacteria in seawater. Results of earlier laborato ry tests show that Greenshipís electrolytic cell needs 2 ñ 3 ppm sodium hypochlorite to achieve a satisfying killing effi ciency in tap water with 35 PSU salt. The test with Greenshipís BWTS was performed with 10 ppm sodium hypochlorite on test water with 10 5 cfu/ml of test organism ( E. coli ) and 35 PSU salt and 3.5 wt% of suspended solids of which 1% consists of Organic Carbon. This proves that sediment concentration is a significant contributor to sodium hypochlor ite demand. Concentration of

sediment in the test water is determined on 3.5 wt% (Total Suspended Solids), which is quite a heavy load of sediment. The concentration sedime nt is three times more concentrated than the Humber, a river known for its high concentration of sediment.

The most important factor to en sure that sodium hypoc hlorite is effectivel y and conservatively applied as a biocide is to ensu re that sediment loads are minimized as much as possible. The Sedimentor minimizes the loads of sediment sign ificantly, which decreases the concentration of sodium hypochlorite needed, to ach ieve the killing efficiency on IMO regulated organisms. This ensures that not more then 10 ppm of sodium hypochlorite is needed. To ensure that enough sodium hypochlorite is used, one could mon itor the residual concentration and keep the concentration between 0.5 and 2.0 ppm.MEPC 57/2/7

ANNEX Page 8

I:\MEPC\57\2-7.doc In the test, no residue of sodi um hypochlorite has been found in the ballast water. Residual measurements on sodium hypochlorite were perf ormed within half an hour after sampling (see table 2.2). These measurements were perf ormed with the ìFree Chlorine Checkî of SenSafeô with a minimal sensitivity of 0.05 ppm. Free chlorine refers to hypochlorite which is responsible for water disinfection and which has not reacted yet w ith the compounds in the water.

Table 2.2: Residual measurements on sodium hypoc hlorite in laboratory test samples

Concentration

sodium hypochlorite produced with

25 A [ppm] Residual measurement on sodium hypochlorite within half an hour after sampling, using Free Chlorine Check of SenSafeô with minimal sensitivity of 0,05 ppm

10,8 Before electrolytic cell After electrolytic cell sample 1 Negative negative sample 2 Negative negative sample 3 Negative negative Blank Negative

Blank: Sample from buffer tank befo re addition of test organism

These tests have been witnessed and the analys es have been reviewed and found in accordance with the test results by Lloydís Register.

The above-mentioned tests were done in the su mmer of 2005. Later test programmes in 2006

and 2007 showed that FAC is effective in the range of 0.5 ñ 3.0 ppm. Further tests for optimization of FAC concentration will be done du ring full-scale tests to prepare the application for Final Approval.MEPC 57/2/7

ANNEX

Page 9

I:\MEPC\57\2-7.doc 2.3 Results of land-based tests

2.3.1 Production of Active Substance and Relevant Chemicals

It is common knowledge, that the production of Free Active Chlorine (FAC) in natural waters

leads to the formation of hal ogenated by-products, like halogena ted methanes and halogenated acetic acids. The applicant has tested the form ation of such by-products during land-based tests of a complete BWMS with natural seawater. Th e results are the basis for the selection of Relevant Chemicals to provide data sets and risk characterization according the requirements of sections 4 and 5 of Procedure (G9).

A BWMS with a capacity of 100m 3 /hr was installed on board of a m oored ship in the harbour of Harlingen, the Netherlands, at the border of the Waddensea in August and September 2007. The BWMS was operated under normal harbour conditions. The 100m 3 /hr is representative for a normal complete system of Greenship. Greenshi p, as the manufacturer of the equipment, can demonstrate by using mathematical modelling and/ or calculations, that any up scaling will not affect the ultimate functioning and effectiveness on board a ship of type and size for which the equipment will be certified.

Samples for chemical analysis were taken and di stributed to different specialized laboratories with full accreditation. Control samples for chemical analysis consisted of seawater treated with the Sedimentor only.

Active Substance

FAC and Total Residual Chlorine (TRC) we re analysed by Omegam-Water, Wognum, the Netherlands. The analyses were performed according to NEN/EN/ISO 7393-2. Results are reported for control samples (TB), samples take n immediately after treatment with FAC (T0) and 30 (T30) resp. 120 (T120) minutes after trea tment. Table 2.3 presents the results.

Table 2.3: Concentrations (mg/l) of FAC and TRC in natural seawater samples

Code: TB Code: T0 Code: T30 Code: T120

TRC mg/l <0.01 2.38 1.83 1.15

FAC mg/l <0.01 1.67 1.25 0.51

Relevant Chemicals

Halomethanes were analysed by Omegam-Laboratoria, Amsterdam, the Netherlands. The analyses of halomethanes were performe d according to NEN/EN/ISO 15680. Results are reported for control samples (3473925), samples ta ken immediately after treatment with Active Substances (3473926), 30 minutes after trea tment (3473927), 12 hours after treatment (3473928) and 24 hours after treatment (3473929). Table 2.4 presents the results.MEPC 57/2/7

ANNEX Page 10

I:\MEPC\57\2-7.doc Table 2.4: Concentrations (μg/l) of chloro- and bromomethanes in treated natural seawater samples

Reference number : 3473925 3473926 3473927 3473928 3473929

Organic compounds ñ chlorinated Volatile chlorinated aliphates: Q dichloromethane μ g/l < 1,0 < 1,0 < 1,0 < 1,0 < 1,0 Q 1,1-dichloroethane μ g/l < 0,5 < 0,5 < 0,5 < 0,5 < 0,5 Q 1,2-dichloroethane μ g/l < 0,5 < 0,5 < 0,5 < 0,5 < 0,5 Q trans-1,2-dichloroethene μ g/l < 0,5 < 0,5 < 0,5 < 0,5 < 0,5 Q cis-1,2-dichloroethene μ g/l < 0,5 < 0,5 < 0,5 < 0,5 < 0,5 Q 1,2-dichloropropane μ g/l < 0,5 < 0,5 < 0,5 < 0,5 < 0,5 Q trichloromethane μ g/l < 0,1 < 0,1 < 0,1 < 0,1 < 0,1 Q tetrachloromethane μ g/l < 0,1 < 0,1 < 0,1 < 0,1 < 0,1 Q 1,1,1-trichloroethane μ g/l < 0,1 < 0,1 < 0,1 < 0,1 < 0,1 Q 1,1,2-trichloroethane μ g/l < 0,1 < 0,1 < 0,1 < 0,1 < 0,1 Q trichloroethene μ g/l < 0,1 < 0,1 < 0,1 < 0,1 < 0,1 Q tetrachloroethene μ g/l < 0,1 < 0,1 < 0,1 < 0,1 < 0,1
sum C/T dichloroethene μ g/l < 0,5 < 0,5 < 0,5 < 0,5 < 0,5
sum chloroalifates μ g/l < 2,1 < 2,1 < 2,1 < 2,1 < 2,1 Volatile chlorinated aliphates ñ remainder:

dibromomethane μ g/l < 0,5 < 0,5 < 0,5 < 0,5 < 1,2
tribromomethane μ g/l < 0,5 200 200 280 290
bromodichloromethane μ g/l < 0,5 1,3 < 0,5 < 0,5 < 0,5
dibromochloromethane μ g/l < 0,5 9,5 9,1 12 13

Haloacetic acids and chloroamines were analysed by RPS Analyse BV, Breda, the Netherlands. The analyses of haloacetic aci ds were performed by GC-MS. Results are reported for control samples (3973470), samples taken immediately after treatment with Active Substances (3973471), 30 minutes after treatm ent (3973472), 12 hrs after tr eatment (3973473) and 24 hrs after treatment (3973474). Table 2.5 presents the results for haloacetic aci ds (see also annex 3). The chloroamines were analysed in another pr ogramme of NHL University for disinfection of

fresh surface water. Salt was added to fresh surf ace water in a concentration of 24 g/l and 10 ppm FAC was produced by the electrolytic cell. The analyses were performed by titration. In the

treated surface water no chloroamines were dete cted with a detection limit of 0.5 mg/l.

Table 2.5: Concentrations (μg/l) of haloacetic acids in treated natural seawater samples

Reference number: 3973470 39734701 3973472 3973473

3973474

Haloacetic acids (Method: GC-MS)
Chloroacetic Acid μ g/l < 1 < 1 < 1 < 1 < 1
Dichloroacetic acid μ g/l < 1 < 1 < 1 < 1 < 1 Trichloorazijnzuur μ g/l < 1 < 1 < 1 < 1 < 1
Bromoacetic Acid μ g/l < 1 < 1 < 1 < 1 < 1
Dibromoacetic acid μ g/l < 1 62 50 64 < 1
tribromoacetic acid μ g/l < 1 < 1 < 1 < 1 < 1
Bromochloroacetic acid μ g/l < 1 1,3 1,6 2,1 < 1
Dibromochloroacetic acid μ g/l < 1 < 1 < 1 < 1 < 1
Bromodichloroacetic acid μ g/l < 1 < 1 < 1 < 1 < 1MEPC 57/2/7

ANNEX Page 11

I:\MEPC\57\2-7.doc Conclusion

The results show, that Active S ubstance (FAC) and Relevant Ch emicals are produced during the disinfection step in the electrolyti c cell. The results are very much in line with the results of other applicants (Japan, Sweden). A typical FAC c oncentration during standard operation of the BWMS is 1.5 ñ 2 ppm immediat ely after treatment, which ra pidly decreases to 0.5 ppm (after 2 hours). Additional measurements, including longer aging of the ballast water will be done during land-based tests for Fi nal Approval with a full-scal e BWMS. Only 2 halogenated

methanes were detected: tribromomethane and dibromochloromethane. Tribromomethane represents more then 96% of the total of the halomethanes and is therefore considered as a Relevant Chemical. Only 2 haloacetic acids were detected: dibromoacetic acid and bromochloroacetic acid. After 24 hours of agin g of the treated ballast water these compounds were not detected anymore. Di bromoacetic acid, representing 97% of the haloacetic acids, is considered a Relevant Chemical. Chapter 3 presen ts the data sets and risk characterization for these compounds.

The formation of hydroxyl radical s by the electrolytic process in the cell, is not considered environmentally relevant, since these radicals have an extremely short half-life in the electrolytic

cell and therefore will never reach the environment.

The formation of small amounts of hydrogen gas (H2) in the electrolytic cell is considered a safety issue and will be dea lt with in chapter 5.

2.3.2 Results of land-based disinf ection tests with seawater

Although the results of laborator y tests on the efficiency of the disinfection process during electrolysis were very convinc ing, it was decided to do supplem ental land-based tests with natural seawater. In the period between 1 May 2007 and 25 June 2007 , five valid replicate test cycles were performed. Each test cycle took pla ce over a period of at least five days. A BWMS with a capacity of 100m 3 /hr was used, which was installed on d eck of a moored barge, which was moored to a tanker, at the quay of the harbour of the city of Harlingen, the Netherlands, at the border of the Waddensea. The BWMS was opera ted under standard conditions. The tanker had

several tanks with normal internal structur es and a total ca pacity of 842 m 3 , resembling ballast water tanks, in which untreated and tr eated seawater could be stored.

Biological and microbiological tests were pe rformed by the Institute f ̧r Umwelttechnik Fachhochschule EUTEC, University of Applied Sciences in Oldenburg, Germany. The results of the five valid replicate test cycles are summarized in table 2.6.MEPC 57/2/7

ANNEX Page 12

I:\MEPC\57\2-7.doc Table 2.6: Summarized results for the land-based test (cycle 1 ñcycle 5, results for untreated uptake water and treated discharge water; the untreated uptake water resembles the harbour water)

Run 1 Run 2 Run 3 Run 4 Run 5

Sizeclass/

IMO regulation untreated uptake treated,

effluent untreated uptake treated,

effluent untreated uptake treated,

effluent untreated uptake treated,

effluent untreated uptake treated,

effluent

1 (plankton

≥ 50 μ m/m3)

IMO: less

than 1 /m3 637000 0 228000 0 706000 0 660000 0 4240000 0

2 (plankton

10-50 μ m/ ml)

IMO: less

than 10/ml 400 0 370 0 600 0 300 7 500 7

3
aerobic heretotrophic

bacteria
cfu/100 ml

NO IMO no test 6000 867000 3500 53000 700 66000 1000 126000 0

3
E. coli (cfu/100 ml)

IMO: less

than

250

cfu/100ml no test 0 73 0 7 0 2 1 9 0

3

Enterococcae
cfu/100 ml

IMO: less

than 100

cfu/100 ml no test 0 0 0 6 0 1 0 8 0

Conclusion

The present results show that the Greenship ba llast water treatment system removes plankton organisms and bacteria from treated ballast water, so that the discharged water complies with the

BWM Convention.

The system was tested in step with the actual practi ce. Harbour water of the port of Harlingen was used as test water, and the test tanker ìAquariusî was moored at a place where much shipping traffic took place, so th e test conditions come close to reality. Within the five cycles,

different free chlorine concentrations were reache d with the electrolysis cell. For best results, a free chlorine production, resulting in a concentra tion of at least 2.6 ppm of free chlorine during uptake should be achieved. Run 4 has shown that ev en smaller free chlorine concentrations lead to sufficient killing rates (see detailed report of run 4), but to be on the safe side, about 2.6 ppm of free chlorine production is r ecommended. Higher free chlorine co ncentrations lead to higher killing rates, but should not be used if not necessary to preserve the environment.MEPC 57/2/7

ANNEX Page 13

I:\MEPC\57\2-7.doc 3 DATA SETS AND RISK CHARACTERIZATION (SEE THE METHODOLOGY, SECTIONS 3 AND 6)

In the previous chapter the performance of th e BWTS and the results of the chemical and (micro)biological analyses in treated ballast wa ter have been reported. The chemical analyses demonstrated the presence of a number of compounds: FAC and Relevant Chemicals (see paragraph 2.3.1) for which an assessment fo r potential envi ronmental risks has to be presented. This chapter presents data for the Active Substance and environmentally Relevant

Chemicals: FAC (in the form of hypochlorite), tribromomethane and dibromoacetic acid. The findings of the chemical analyses will be valid ated against the literature data on toxicity of these compounds. The conclusions per individual compound can never exclude adverse environmental effects of discharged treated balla st water. Therefore w hole effluent toxicity testing was performed as well a nd is reported in chapter 4.

The applicant has used the key data summ ary form as presented in the Methodology, for the content in this chapter, the applicant has us ed the following main sources of information:

Risk assessment report for sodium hypoc hlorite, version January 2007, including annexes 2, 3 and 4, provided by: Dipartimen to di Ambiente e Connessa Prevenzione Primaria, Istituto Superiori di Sanita, Rome, Ital y (reference 33).

This document is a final draft to become a full EU Risk Assessment Report (RAR) and is produced by Member State Italy w ith support of the European chlo rine industry. An nex 3 in this reference gives a full summary of literature data on eco-toxicology study results for fish, invertebrates and algae in brackish and seawater:

- MEPC 56/2: Application by Japan fo r Basic Approval of Active Substances

- Annex 7 to MEPC 56/2/2: Review by the GESAMP-BWWG of the application by Japan (MEPC56/2), appendices 1, 3 and 5

- MEPC 55/2/4: Application by Sweden for Basic Approval of Active Substances

- Annex 7 to MEPC 55/2/16: Review by the GESAMP-BWWG of the application by Sweden (MEPC 55/2/4)

Chemical identification

For three compounds, a complete data set and risk characterization will be presented. Chapter 2, paragraph 2.3.1, presents the list of all possible by-products. In th e conclusion, it is motivated why data sets and risk assessments for three co mpounds will be presented in this application for Basic Approval. The Active Substance (FAC), th e Relevant Chemicals and all other possible byproducts, including bromate, will be analysed in the land-based tests, n ecessary for application for Final Approval.

Composition of Preparation Component Chemical Name CAS Number Concentration (%) AS, RC or Other Sodium-hypochlorite 7681-52-9 Max 1,67 ppm AS Tribromomethane 75-25-2 Max 290 ppb RC 2,2 Dibromoacetic acid 631-64-1 Max 64 ppb RCMEPC 57/2/7

ANNEX Page 14

I:\MEPC\57\2-7.doc 3.1 Free active chlorine (hypochlorite)

Since FAC is present as hypochlor ite in seawater (pH = 8 to 8.2), the data set and risk characterization will be presented for hypochlorit e. The FAC is interpreted as Total Residual Oxidants (TRO).

3.1.1 Effects on aquatic organisms

Acute aquatic toxicity

Species Effect Value (mg/L) References

F Oncorhynchus kisutch 96 h LC 50 0.032

C Ceriodaphnia dubia 24 h LC 50 0.005

A Thalassiosira pseudonana 24 h IC 50 0.075

M Crassostrea virginica 96 h EC 50 0.023 RAR Hypochlorite (Ref. 33); Thatcher, 1978; Taylor, 1993; Gentile et al. , 1979; Robertd and Gleeson, 1978

F = Fish, C = Crustacea, A = Algae, M = Mollusc

Chronic aquatic toxicity

Species Effect Value (mg/L) References

F Menidia peninsulae 28d NOEC 0.04

Chlorineproduced

oxidant

C Pandalus danae 28d NOEC 0.05 mg/l Free

Active Chlorine

A Unknown species 7d NOEC 0.0027 mg/l

TRO

M Crassostrea virginica 15d NOEC 0.007 mg/l

TRO RAR Hypochlorite (Ref. 33); Goodman, 1983; Gibson, 1976; Cairns, 1990; Liden,

1980.

F = Fish, C = Crustacea, A = Algae, M = Mollusc

Information on Endocrine Disruption

No reports were found, indicati ng that FAC might have any endocrine effects on aquatic organisms.

Sediment toxicity/bioavailability/biomagnification/bioconcentration/foodweb-population effects

The Log Pow value of hypo chlorous acid is -0.87 (estimated) a nd that of sodium hypochlorite ñ 3.42 (estimated), which is smaller than the crite ria of Log Pow=3. The Koc value of sodium hypochlorite is 14.3 l/kg (e stimated by EPI Suite TM ), which is smaller than 500 l/kg.

Therefore, FAC is not considered to absorb to sediment or to accumulate in the food web (see also results of MAM-PEC).MEPC 57/2/7

ANNEX Page 15

I:\MEPC\57\2-7.doc 3.1.2 Mammalian toxicity

Acute mammalian toxicity

Exposure Route Value Species References

Oral LD 50 (mg/kg) 850 rat WHO 2000 (Calcium hypochlorite

Dermal LD 50 (mg/kg) > 10000 rabbit MEPC 57/2/1

Inhalation LC 50 (mg/l/4h) > 10.5 rat MEPC 57/2/1

Corrosion/irritation

Species Method Results (including scores where available) References

Skin Rabbit OECD 404 2% slight (1.2) 20% severe (5.2) IUCLID 2000

Eye Rabbit 5.25% irritant 0.5% slight irritant

0.05% NOEC Pashley, 1985

Sensitization

A 2.8% solution of sodium hypochlorite solution was not found to be a skin sensitizer in guinea pigs (IUCLID 2000).

Repeated dose toxicity

Exposure Route Oral Exposure Duration 90 days Exposure dose 200 mg/l TRO in drinking water Species Mouse

Method

Results

NOAEL 50 mg/l (10-12 mg/kg/day)

NOEL

Reference Daniel et al . (1991)MEPC 57/2/7

ANNEX Page 16

I:\MEPC\57\2-7.doc Chronic and long-term toxicity

Exposure Route Oral Exposure Duration 2 years Exposure dose Up to 24.2 mg/kg/day (14.4 in rats, 24.2 in Mouse) Species Rat and Mouse

Method

Results

NOAEL 14.4 mg/kg/day

NOEL

Reference NTP, 1992

Development and reproductive toxicity

Exposure Route Oral Exposure Duration 7 generations Exposure dose 100 mg TRO/l (drinking water treated with sodium hypochlorite) Species Rat

Method

Results

NOAEL 14.4 mg/kg/day

NOEL

Reference Druckrey, 1968

Carcinogenicity/mutagenicity

Species Method

Details Results Reference

Carcinogenicity Not carcinogenic NTP, 1992

Mutagenicity Not mutagenic Hayashi, 1988; Meier et al.

, 1985

3.1.3 Environmental fate

Environmental fate

Taylor (1993) measured the decay rate of hypoc hlorous acid and hypochlorite ions in an acute aquatic toxicity test. The decay rate was reported as follows: the half-life at pH 7.0 (main formula is HClO) is about 0.8 hour and the half -life at pH 8.0 (main formula is ClO - ) is about 1.4 hour. The A.I.S.E. document (ref.1) repor ts a half-life of 45-60 minutes.

In the presence of organic material, FAC disappear s very quickly, due to its oxidizing property. Therefore, the resulting TRO in th e receiving water will be very low (see also annex 2 to RAR, reference 33).MEPC 57/2/7

ANNEX Page 17

I:\MEPC\57\2-7.doc Partition coefficients

Method Results Reference Log Pow estimated - 3.42 EPI Suite TM

Koc estimated 14.3 l/kg EPI Suite TM

Reaction with organic matter

It is known that when FAC reacts with organic material in water, by-products such as halomethanes and halogenated acetic acids are formed. Four substances (tribromomethane, dibromochloromethane, 2,2-dibromoacetic acid a nd bromochloroacetic acid) were detected by preliminary experiments as by-products in treated ballast water (see chapter 2).

Potential physical effects on w ildlife and benthic habitats

FAC is highly soluble in water (IUCLID, 2000) a nd does not float on the surface or sediment to the bottom. So, the compound is not subject to eval uation of potential physical effects on wildlife

and benthic habitants under the GESA MP hazard evaluation (GESAMP, 2002).

Potential residuals in seafood

No report concerning persistence of FAC in seafood is available. As FAC has a low bioaccumulation potential, its persisten ce in seafood is assumed to be low.

Any known interactive effects

Free active chlorine may genera te corrosive gas including gaseous chlorine under high temperature or low pH condition (<pH 4). These conditions will not occur in the environment or

in the ballast water tanks.

3.1.4 Physical and chemical properties

Physical property Sodium hypochlorite

CAS number 7681-52-9 Molecule weight 74.44 Melting point (°C) - 25 Boiling point (°C) 107 Flash point (°C) Density (kg/dm 3 ) at 25 °C 1.23*10 3
Vapour pressure @ 20°C (Pa) 1740-2000 Water solubility (g/L) miscible(293) pH in solution 14 Octanol/water partition coefficient (log Pow) -0.87

Dissociation constant (pKa) 7.63 Oxidation reduction potential 1.7 Oxidizing properties Strong oxidizing agent Viscosity (mPa.s) 2.6MEPC 57/2/7

ANNEX Page 18

I:\MEPC\57\2-7.doc PH, salinity, TSS, POC, DOC

The pH and salinity of the treated ballast water is the same as the natural values of the uptake water. The pH varies from 7.4 to 8.0. Salinity varies from 22 ñ 34 PSU. TSS, DOC and POC were analysed in samples taken during the te st on killing rates in the summer of 2007. All parameters vary highly due to conditions in the harbour th rough tidal movements and ship movements. TSS varied between 20 and 100 mg/l. POC values were in the range 0.5 ñ 29 mg/l and DOC values were in the range of 1.0 ñ 52 mg/l.

3.1.5 Other information

Analytical Methods for Measuring the Co ncentration at environmentally Relevant

Concentrations

Method Fotometric (DPD-1) Applicability salt water Sensitivity < 0.01 mg/l

Reference NEN/EN/ISO 7393-2

Material Safety Data Sheet provided: no

GHS Classification: no

3.1.6 Risk characterization

Persistent (y/n) Bioaccumulative (y/n) Toxic (y/n) Reference

No no yes reference 31 and this document

Evaluation of the treated ballast water

See chapter 2 ñ for the chemical evaluation of th e treated ballast water and chapter 4 ñ for the preliminary ecotoxicological evaluation.

Prediction of discharge and environmental concentrations: MAM-PEC

For the calculation of the Pred icted Environmental Concentrati on (PEC), the model MAM-PEC, version 2.0, was applied. This model is freely available and can be downloaded. The model has been developed by Delft Hydraulics (now DE LTARES) and the Free University, Amsterdam. The model was developed to calculate leaching of anti-fouling paints from ships. It is based on the assumption that a permanent source of the pollut ant is present in the ha rbour and calculates a steady state concentration of the pollutant. Calculations can be done for different harbours with different dimensions and ìsurroundingsî, whic h in practice means the channel to which the harbour is connected. It also has the possibility for calculations in a shipping lane of 200 km 2

. In this application, a standard OECD-EU harbour has been used with an exchange volume of 51,117,000 m 3 per tidal period of 12.1 hours. The daily volume of discharged treated ballast

water and the concentration of the Active Substa nce and Relevant Chemi cals are critical input data. Reliable data on daily discharge of ballast wa ter is difficult to obtain. Other applicants have used volumes varying between 396,000m 3 /day (Korea) and 50,000 m 3 /day (Sweden).MEPC 57/2/7

ANNEX Page 19

I:\MEPC\57\2-7.doc According to the authorities of the Port of Rotte rdam, the real value will be closer to 50,000 than to 396,000. It depends on the type of harbour: in Rotterdam the intake of ballast water is higher than the discharge, since ships are coming loaded. A port that handles mainly tankers that arrive unloaded, may have a higher daily di scharge than the Port of Rotterdam. This applicant has used the figure of 200,000 m 3 /day. The most important input data is mentioned as follows:

Main input data for FAC is:

Load: 334 kg/day (based on 200,000m 3 /day and maximum of 1.67 mg/l)

Half-life : 0.041 days in water

The MAM-PEC model calculates fo r FAC a steady state situati on (= PEC) in the harbour

of 0.34 μ g/l and of 0.017 μ g/l in the surroundings. The mode l calculates a steady state situation in the shipping lane of 0.014 μ g/l.

Effects on aquatic organisms: predic ted-no-effect concentrations (PNEC)

In various documents, the predicted no effect concentrations of FAC (PNEC) have been

calculated with different outcomes. Annex 7 to MEPC 56/2/2 (review by the GESAMP-BWWG of the application by Japan, MEPC 56/2), reports a value of 0.07 μ g/l. Reference 33 reports a

value of 0.04 μ g/l. Document MEPC 57/2/1 (a pplication by Korea) reports 8 μ g/l.

3.1 Tribromomethane

3.2.1 Effects on aquatic organisms

Acute aquatic toxicity

Species Effect Value (mg/L) Calculated

NOEC (mg/L) References

F Cyprinodon variegates 96 h LC 50 7.1 2.9

C Daphnia magna 24-48 h LC 50 46 <7.8

A Skeletonema costatum 96 h EC 50 12.3 Ward et al ., 1981;
Heitmuller et al ., 1981; Ecotox, 2005; LeBlanc, 1980 F = Fish, C = Crustacea, A = Algae

Chronic aquatic toxicity

Species Effect Value Reference

F Cyprinodon variegatus 28d NOEC 4.8 mg/l

C

A Ward et al ., 1981

Information on endocrine disruption

No reports were found, indicating that tribromo methane might have any endocrine effects on aquatic organisms.MEPC 57/2/7

ANNEX Page 20

I:\MEPC\57\2-7.doc Sediment toxicity/bioavailability/biomagnificatio n/bioconcentration/food web-population effects

The Log Pow value of tribromomethane is 2.4 (ATD SR, 2005), which is smaller than the criteria of Log Pow = 3. The Koc value of tribromome thane is 114.8 l/kg (annex 7 to MEPC 56/2/2), which is smaller than 500 l/kg. Therefore, tribro momethane is not considered to absorb to

sediment or to accumulate in the food web.

3.2.2 Mammalian toxicity

Acute mammalian toxicity

Exposure Route Value Species References

Oral LD 50 (mg/kg) 1147 Rat Chu et al ., 1980

Dermal LD 50 (mg/kg)

Inhalation LC 50 (mg/l/4h) >7000 ppm/1 h Dog MEPC 56/2, paragraph 2.3.2.2 Sub-acute toxicity NOAEL oral (mg/kg bw/day) 2 weeks

Corrosion/irritation

Species Method Results (including

scores where available) Reference

Skin Rabbit Moderately irritating Eye Rabbit Moderately irritating Respiratory system Moderately irritating Torkelson et al ., 1994

Sensitization

No sensitization data are reported for tribromomethane.MEPC 57/2/7

ANNEX Page 21

I:\MEPC\57\2-7.doc Repeated dose toxicity

Exposure Route Oral Exposure Duration 13 weeks Exposure dose 50 mg/kg/day Species Rat

Method

Results

NOAEL 25 mg/kg/d

NOEL

Reference NTP, 1989

Chronic and long-term toxicity

Exposure Route 1 year Exposure Duration Exposure dose 6.2-57 mg/kg/d (Rat) 8.3-73 mg/kg/d (Mouse) Species Rat and Mouse

Method

Results

NOAEL <6.2 mg/kg/d

NOEL <6.2 mg/kg/d

Reference Moore et al ., 1994

Development and reproductive toxicity

Exposure Route Exposure Duration Not given for rat study 105 d for Mouse study Exposure dose 50-200 mg/kg Species Rat and Mouse

Method

Results foetotoxicity observed in rats No fertility effects seen in Mouse study

NOAEL

NOEL

Reference WHO, 2000MEPC 57/2/7

ANNEX Page 22

I:\MEPC\57\2-7.doc Carcinogenicity/mutagenicity

Species Method

Details Results References

Carcinogenicity Human Probable

Human Carcinogen U.S. EPA (IRIS)

Mutagenicity Salmonella.

Mammalian

cells Ames test Mixed

Mixed Simmon et al ., 1978;
Simmon et al ., 1979 ; NTP, 1989 ; Rapson
et al ., 1980 ; Varma
et al. , 1988 ; Morimoto, 1983 ; Fuji et al ., 1993 ; Galloways et al ., 1985

3.2.3 Environmental fate

Modes of degradation (abiotic) and persistence

The rate constant of hydrolysis for tribromomethane is estimated at 3.2x10 -11 /sec (Mabey, W. et al., 1978). It thus is a stable compound in water with an estimated half-life of 686 years.

Partition coefficients

Method Results Reference Log Pow observed 2.4 CITI, 1992; Sangster, 1994

Koc observed 114.8 l/kg Annex 7 to MEPC 56/2/2

Reaction with organic matter

No reports on the reaction of tribromo methane with organic matter were found.

Potential physical effects on w ildlife and benthic habitats

Tribromomethane has a higher specific gravity than seawater (2.890 at 20°C) and a low solubility (3.1 g/l (20°C)). Therefore, according to an evaluation based on the GESAMP hazard evaluation (GESAMP, 2002), the compound is classified as Ra ting S (substances havi ng a potential effect on benthic habitats).

Potential residues in seafood

Tribromomethane has a low bioaccumulation potentia l and therefore it is unlikely that residues

will be found in seaf ood (see paragraph 3.2.1).MEPC 57/2/7

ANNEX Page 23

I:\MEPC\57\2-7.doc 3.2.4 Physical and chemical properties

Molecular formula CHBr3 Molecular weight 252.73

CAS-number 75-25-2 Flammability Non-flammable Vapour pressure (Pa) at 20 oC 534 Water Solubility (mg/L) Slightly soluble

3.2.5 Other information

Analytical method for measuring the co ncentration at environmentally relevant

concentrations

Method GC-MS Applicability salt water Sensitivity < 0.5 ñ 0.1 μ g/l

Reference NEN/EN/ISO 15680

Material Safety Data Sheet provided: no

GHS Classification: no

3.2.6 Risk characterization

Persistent (y/n) Bioacumulative (y/n) Toxic (y/n) Reference

Yes no no this document

Evaluation of the treated ballast water

See chapter 2 ñ for the chemical evaluation of th e treated ballast water and chapter 4 ñ for the preliminary ecotoxicological evaluation.

Prediction of discharge and environmental concentrations: MAM-PEC

For the calculation of the Pred icted Environmental Concentrati on (PEC), the model MAM-PEC, version 2.0, was applied. This model is freely available and can be downloaded. The model has been developed by Delft Hydraulics (now DE LTARES) and the Free University, Amsterdam. The model was developed to calculate leaching of anti-fouling paints from ships. It is based on the assumption that a permanent source of the pollut ant is present in the ha rbour and calculates a steady state concentration of the pollutant. Calculations can be done for different harbours with different dimensions and ìsurroundingsî, whic h in practice means the channel to which the harbour is connected. It also has the possibility for calculations in a shipping lane of 200 km 2

. In this application, a standard OECD-EU harbour has been used with an exchange volume of 51,117,000 m 3 per tidal period of 12.1 hours. The daily volume of discharged treated ballast

water and the concentration of the Active Substa nce and Relevant Chemi cals are critical input data. Reliable data on daily discharge of ballast wa ter is difficult to obtain. Other applicants have used volumes varying between 396,000 m 3 /day (Korea) and 50,000 m 3 /day (Sweden). According

to the authorities of th e Port of Rotterdam, the real value will be closer to 50,000 than to 396,000. It depends on the type of harbour: in Rotterdam the intake of ba llast water is higher than theMEPC 57/2/7

ANNEX Page 24

I:\MEPC\57\2-7.doc discharge, since ships are coming loaded. A por t that handles mainly tankers that arrive unloaded, may have a higher daily di scharge than the Port of Rotterdam. This applicant has used the figure of 200,000 m 3 /day. The most important input data is mentioned as follows:

Load: 58 kg/day (based on 200.000 m 3 /day and maximum of 0.290 mg/l)

Half-life: 340 days in sediment

The MAM-PEC model calculates for tribromomethan e a steady state situation (= PEC) in the

harbour of 1.28 μ g/l and of 0.036 μ g/l in the surroundings. The m odel calculates a steady state situation in the shipping lane of 0.005 μ g/l.

Effects on aquatic organisms: predic ted-no-effect concentrations (PNEC)

In various documents, the predicted no effect c oncentrations of tribromomethane (PNEC) have

been calculated with different outcomes. Annex 7 to MEPC 56/2/2 (review by the GESAMP-BWWG of the application by Japa n, MEPC 56/2), repo rts a value of 7.1 μ g/l. Reference 33 reports a value of 146 μ g/l. RIVM (2005) reports a value of 1. 5 μ g/l.

3.2 2,2 Dibromoacetic Acid

3.3.1 Effects on aquatic organisms

Acute aquatic toxicity data

Species Effect Value Reference

F Pimephales promelas 96 h LC 50 6.9 mg/l Mayes et al , 1985

C

A F = Fish, C = Crustacea, A = Algae

Chronic aquatic toxicity

No data is available on the chronic a quatic toxicity of 2,2-dibromoacetic acid.

Information on endocrine disruption

No reports were found, indicating that 2,2 dibrom oacetic acid might have any endocrine effects on aquatic organisms.

Sediment toxicity/bioavailability/biomagnificatio n/bioconcentration/food web-population effects

The Log Pow value of 2,2 dibromoacetic acid is 1.22 (observed), which is smaller than the criteria of Log Pow = 3. The Koc value of 2,2 dibromoacetic is 1.90 l/ kg (), which is smaller than 500 l/kg.

Therefore, 2,2 dibromoacetic acid is not considered to absorb to sediment or to accumulate in the

food web.MEPC 57/2/7

ANNEX Page 25

I:\MEPC\57\2-7.doc 3.3.2 Mammalian toxicity

Acute mammalian toxicity

Exposure Route Value Species Reference

Oral LD 50 (mg/kg) 1737 Rat Linder et al ., 1994

Dermal LD 50 (mg/kg)

Inhalation LC 50 (mg/l/4h)

Corrosion/irritation

Species Method Results (including scores where available) Reference

Skin Corrosive Eye Severe Lung Severe Assumed as a strong acid

Sensitization

No sensitization data are repor ted for 2,2 dibromoacetic acid.

Repeated dose toxicity

Exposure Route Oral Oral Exposure Duration 28d 3 months Exposure dose 0-2000 mg/l Species Mouse Mouse

Method

Results No deaths but various effects seen at >500 mg/l

NOAEL 38 mg/kg/d Not reported

NOEL Not reported

Reference WHO, 2004

Chronic and long-term toxicity

Exposure Route Oral Oral Exposure Duration 2 years 2 years Exposure dose 0-1000 mg/l Species Mouse Rat

Method

Results Increased body weights at 50 and 500 mg/l in males only Decrease body weights at 500 and 1000 mg/l

NOAEL Not reported Not reported

NOEL Not reported Not reported

Reference NTP, 2005MEPC 57/2/7

ANNEX Page 26

I:\MEPC\57\2-7.doc Development and reproductive toxicity

Exposure Route Oral Exposure Duration Exposure dose 0-250 mg/kg/d Species Rat

Method

Results Increased E2 at 250 mg/kg/d

NOAEL 125 mg/kg/d

NOEL

Reference Cummings et al . (1998)

Carcinogenicity/mutagenicity

Species Method

Details Results Reference

Carcinogenicity Mouse and

Rat Carcinogenic (tumours/cancers observed) NTP, 2007

Mutagenicity Salmonella typhimurium Ames

test Positive at 10 mg/l NTP, 2007; Giller

et al. , 1997

3.3.3 Environmental fate

Environmental fate

Holmes et al. (2001) measured the decay rate of 2,2 dibr omoacetic acid with in vivo and vitro toxicity tests. They repor t a half-life of 1.5 hours.

Partition coefficients

Method Results Reference Log Pow estimated 1.22 EPI Suite TM

Koc estimated 1.90 l/kg EPI Suite TM

Reaction with organic matter/ Potential physical effects on wildlife and benthic habitats / Potential

residuals in seafood

The 2,2 Dibromoacetic acid is highly soluble in water; actually it is highly hygroscopic.

It therefore will not react substant ially with organic matter nor will it precipitate in wildlife or

benthic habitats or accumulate in seafood.

3.3.4 Physical and chemical properties

Molecular formula CHBr2COOH Molecular weight 217.860

CAS-number 631-64-1 Flammability Non-flammable Vapour pressure (Pa) at 25 oC 0.023 Water Solubility (mg/L) Highly solubleMEPC 57/2/7

ANNEX Page 27

I:\MEPC\57\2-7.doc 3.3.5 Other information

Analytical methods for measurin g the concentration at environm entally relevant concentrations

Method GC-MS Applicability salt water Sensitivity < 1 μ g/l

Reference RPS Laboratories: Halacet_V1 methodology

Material Safety Data Sheet provided: no

GHS Classification: no

3.3.6 Risk characterization

Persistent (y/n) Bioacumulative (y/n) Toxic (y/n) Reference

No no no this document

Evaluation of the treated ballast water

See chapter 2 ñ for the chemical evaluation of th e treated ballast water and chapter 4 ñ for the preliminary ecotoxicological evaluation.

Prediction of discharge and environmental concentrations: MAM-PEC

For the calculation of the Pred icted Environmental Concentrati on (PEC), the model MAM-PEC, version 2.0, was applied. This model is freely available and can be downloaded. The model has been developed by Delft Hydraulics (now DE LTARES) and the Free University, Amsterdam. The model was developed to calculate leaching of anti-fouling paints from ships. It is based on the assumption that a permanent source of the pollut ant is present in the ha rbour and calculates a steady state concentration of the pollutant. Calculations can be done for different harbours with different dimensions and ìsurroundingsî, whic h in practice means the channel to which the harbour is connected. It also has the possibili ty for calculations in a shipping lane of 200 km 2 . In this application, a standard OECD-EU harbour has been used with an exchange volume of 51,117,000 m 3 per tidal period of 12.1 hours. The daily volume of discharged treated ballast

water and the concentration of the Active Substa nce and Relevant Chemi cals are critical input data. Reliable data on daily discharge of ballast wa ter is difficult to obtain. Other applicants have used volumes varying between 396,000 m 3 /day (Korea) and 50,000 m 3 /day (Sweden). According

to the authorities of th e Port of Rotterdam, the real value will be closer to 50,000 than to 396,000. It depends on the type of harbour: in Rotterdam the intake of ba llast water is higher then the discharge, since ships are coming loaded. A por t that handles mainly tankers that arrive unloaded, may have a higher daily di scharge than the Port of Rotterdam. This applicant has used the figure of 200,000 m 3 /day. The most important input data is mentioned as follows:

Load: 13 kg/day (based on 200.000 m 3 /day and maximum of 0.064 mg/l)

Half-life: 0.0625 day in waterMEPC 57/2/7

ANNEX Page 28

I:\MEPC\57\2-7.doc The MAM-PEC model calculates for 2,2 dibromoacetic acid a steady state situation (= PEC) in

the harbour of 0.019 μ g/l and of 0.001 μ g/l in the surroundings. The model calculates a steady

state situation in th e shipping lane of 0.0007 μ g/l.

Effects on aquatic organisms: predic ted-no-effect concentrations (PNEC)

In various documents, the predicted no effect c oncentrations of 2,2 dibromoacetic acid (PNEC)

have been derived with different outcome s. Annex 7 to MEPC 56/2/2 (review by the GESAMP-BWWG of the application by Japa n, MEPC 56/2), repo rts a value of 6.9 μ g/l. Reference 33 reports a value of 0.72 μ g/l.

4 TOXICITY EVALUATION OF THE TREATED BALLAST WATER

In order to comply with paragraph 5.2 of Procedur e (G9) regarding toxicity testing of the treated ballast water, the applicant has commissioned the Institute for Marine Re sources ands Ecosystem Studies of Wageningen IMARES, Den Helder, Th e Netherlands, to perform the prescribed toxicity tests on whole effluent of the BWTS (treated ballast water). The samples were taken under the same technical conditions as samples taken for chemical analysis (see paragraph 2.3 of this document). This chapter presents the full report on the results of the tests.

Summary

Greenship B.V. has developed a ballast water ma nagement system consisting of a Sedimentor (multi-hydrocyclone system) and an electroly tic cell, producing hypochlorite for killing organisms. The residual effects of seawater treated with the electrolytic cell was investigated with bioassays, using three ma rine organisms: an algae ( Skeletonema costatum ) a crustacean ( Artemia franciscana ) and a fish ( Solea solea ).

The tests were performed at the laboratory of IMARES in Den He lder, using treated ballast water aged for 24 h, 48 h, 72 h and 96 h. Each test day dilutions of 10% 18 %, 33%, 56% and 100% treated ballast water were made, using untreated ballast water (only hydrocyclone treated) as dilution medium and blank. The salinity of the te st water was adjusted to approximately 30% by adding NaCl.

Treated ballast water showed no ne gative effects in the tests with the crustacean and the fish.

In the algal growth inhi bition test, significant nega tive effects of treated ba llast water were found. The algae did not survive exposure to 100% treated ballast water in any of the test runs. Exposure
to a dilution of 56% treated ballast water resu lted in complete mortality in the test run with 24 h aged ballast water, but recovery was observed in olde r ballast water. In 96 h aged ballast water, full recovery was observed dur ing the course of the test (3 days).

4.1 Introduction

Ballast water is used by ships to compensate for differences in cargo load. With the ballast water, non-indigenous organisms are tr ansported worldwide from ships transporting cargo between regions, which may result in disturbances of re gional ecosystems. IMO has set out a mandatory

framework for treatment of ballast water on board of ships, in order to minimize the risk of ballast water discharges. Bioassays have to be conducted to assess the eff ects of treated ballast water on the receiving ecosystems.MEPC 57/2/7

ANNEX Page 29

I:\MEPC\57\2-7.doc Greenship B.V. has developed a Ballast Water Management System consisting of a Sedimentor (multi-hydrocyclone system) and an electroly tic cell, producing hypochlorite for killing organisms. The residual effects of seawater treated with the electrolytic cell was investigated with bioassays, using three marine organisms: an algae ( Skeletonema costatum ), a crustacean ( Artemia franciscana ), and a fish ( Solea solea ) ñ one to four days after the treatment.

4.2 Materials and methods

4.2.1 General

On Tuesday, 21 August 2007, a treatment ru n was performed in Harlingen Harbour, The Netherlands. From IMARES, A.C. Sneekes and J. Cuperus were present to collect the water samples. The Treated Ballast Water (TBW) was seawater from Harlingen Harbour, treated with the hydrocyclone and the electrolyt ic cell. It was collected in four 25 L PE jerry cans, coded TBW1 to TBW4. A 5th jerry can was taken as back-up. The Untreated Ballast Water (UBW), was taken at the same location in Harlingen Harb our, but was only treated with the hydrocyclone.
The UBW was collected in four 25 L PE jerry cans, co ded UBW1 to UBW4 and a 5th as back-up.

After filling the last jerry can, the water samples were directly transported to the laboratory of

IMARES in Den Helder and stored in the dark at approx. 16°C.

4.2.2 Bioassays

Bioassays were initiated approximately 24 h (22 August), 48 h (23 August), 72 h (24 August) and 96 h (25 August) after the water was collec ted and treated in Harlingen Harbour. As the oxygen content of the water was too low for the tests, before test initiation, the water used for the tests was aerated overnight. Before use, water quality parameters were measured in the jerrycans. As the salinity of the water in Harlingen Harbour was too low for the test organisms (approximately 19%), 10 L was tapped into a large bottle and the salinity was adjusted with NaCl (p.a.-grade) to approximately 29% . After mixing for at least 30 mi nutes water quality parameters were measured again.

Each test day, dilution series were made by mixing TBW with UB W in the following proportions (see table 4.1):

Table 4.1: Dilutions of treated ballast water (TBW ) with untreated ballast water (UBW)

Code TBW UBW

BW0 0% 100%

BW1 10% 90%

BW2 17.8% 82.2%

BW3 32.6% 67.4%

BW4 56.2% 43.8%

BW5 100% 0%

The dilution series were made directly from st ored water (approximately 16°C) and was then set away at the conditions of different bioassay s for acclimatization. Water parameters were

measured in the dilution series for Skeletonema costatum and Solea solea at start and termination

of the test. The test water for Artemia franciscana was taken from the dilution series of the fish

test.MEPC 57/2/7

ANNEX Page 30

I:\MEPC\57\2-7.doc 4.2.2.1 Skeletonema costatum

The test protocol is based upon OECD 201 (1984) a nd ISO10253 (2006). The test species used was

the marine diatom Skeletonema costatum . An active growing culture was obtained from TNO Industry (J. Klijnstra). With this inoculum, cultures were started in F2-medium, incubated under the conditions of the test. In the temperatur e-controlled shaking-cabi net normally used for algal-growth tests, only one test series (including reference test) could be placed. Therefore, part of the tests was performed in a climate chamber. In this location, the te st flasks had to be shaken by hand several times a day to facilita te gas exchange. The control grow th rate on this location was actually too slow (les s than 16x multiplication in 72 h), due to a slightly lower temperature (18°C vs 20°C), a different light source (cool white vs fl uoro) and the lack of continuous shaking.

Therefore, test series I (24 h after treatment) and IV (96 h after treatment) were performed in the shaking cabinet, while only test se ries II (24 h) and III (72 h) were performed in the climate

chamber.

The tests were performed in 250 mL Erlenmeyer flask, filled w ith approximately 150 mL test medium. For the algal growth test, the TB W was diluted with UBW. The UBW was 0.2μm filtered in order to remove the natural phytopla nkton. Each concentration was tested in 3-fold. To each concentration, the same amount of F2-growth medium was added.

Each test concentration was inoculated with al gae from a log-phase gr owing culture. The start concentration was approximately 20 μg/L chlorophy ll-a, as measured with a fluorometer before filling the replicate flasks. The flasks were in cubated in continuous light at 18°C or 20°C.

The development of the algal biomass, measured as chlorophyll-a concentration, was measured

each 24 h.

With each test series, a reference test was conducted, using potassiumdichromate (K 2 Cr 2 O 7 ) as

toxicant.MEPC 57/2/7

ANNEX Page 31

I:\MEPC\57\2-7.doc Summary of test conditions for th e algal growth inhibition test

with the marine diatom Skeletonema costatum

Test organism Skeletonema costatum Test organism

source TNO Industry, continuous culture

Test organism life
stage Log-phase growth, ca. 4 day old culture

Test duration 72 h Test chamber 250 mL Erlenmeyer

Test solution

volume 200 mL per replicate

Initial

concentration Approx. 20 μg/L chlorophyll-a

Replicates 3

Method Fluorescence measurements Endpoint Growth (as con centration chlorophyll-a) Test temperature 20°C (series I and IV) 18°C (series II and III) Dilution water UBW, 0.2 μm filtered/autoclaved

Media stock

solutions F2-medium

Photoperiod Permanent light Type of light Fluoro (series I and IV) Cool white (series II and III) Shaking 130 rpm (series I and IV) by hand, several times a day (series II and III) Test protocol OECD 201 (1984) and ISO10253 (2006)

Control acceptability

criteria Minimum chlorophyll-a increase by factor 16

after 72 h

Reference test K 2 Cr 2 O 7

Statistics

Growth rates were calculated by fitting an exponential growth curve through the individual samples (replicates). The curve was fitted with the statistical program GraphPad Prism (v 4.03, Jan. 2005), using the following formula:

0 0 ) ln( ) ln(

t t N N μ

L L

− −

=

Wherein: μ = specific growth rate (day -1 )

N L = chlorophyll-a concentration at t L (at 24, 48, or 72 h).

N 0 = chlorophyll-a concentration at t 0 (initial concentration)

t L = time of measurements (days)

t 0 = start of measurements (0 days).MEPC 57/2/7

ANNEX Page 32

I:\MEPC\57\2-7.doc EC 50 s were calculated as being the concentration/ dilution that causes 50% growth inhibition, using the formula for a dose-response curve with variable slope built-in in GraphPad Prism. GraphPad Prism uses the following formula:

Hillslope x EC

Bottom Top Bottom y
* ) 50 (log 10 1

−

+ − + =

Wherein:

x = Concentration or dilution y = Specific growth rate μ Top = Maximum growth rate Bottom = Minimum growth rate (0)

Bottom was set at a fixed value of 0, to correct for negative growth (mortality).

The NOEC was determined as the highest test c oncentration in which the growth rate was not significantly reduced compared to the control (U BW). This was assessed by means of a one-way ANOVA followed by Dunnettís multiple comparison post-test.

4.2.2.2 Artemia franciscana

The test was performed in a multiwell testplate us ing instar II-III larvae of the brine shrimp Artemia franciscana , obtained as dried cysts from Microl an B.V., The Netherlands, conform the Standard Operational Procedure for AR TOXKIT Mô (MicroBioTests Inc. V100603).

The cysts were incubated in artificial seawater (s alinity 35%) 30 h prior to the start of the tests

at 25°C with continuous illumination. After 30 h, most cysts were hatched and the larvae developed into instar II-III stages.

Approximately 50 larvae were tr ansferred to a rinsing well cont aining test medium. From the rinsing well, 10 larvae were transferred to the th ree test wells per concentration/dilution for the test. The well plate was incubated at 25°C in th e dark. After 24 h living and dead larvae in the

test wells were counted.

With each test series, a reference test was conducted, using potassiumdichromate (K 2 Cr 2 O 7 ) as

toxicant.MEPC 57/2/7

ANNEX Page 33

I:\MEPC\57\2-7.doc Summary of test conditions fo r the ARTOXKIT M with the brine shrimp Artemia franciscana

Test organism Artemia franciscana Test organism source ARTOXKI T M, MicroBioTests Inc. Test organism life stage Larvae <24 h old

Test duration 24 h Test chamber 24-well plate Test solution volume 1 mL per replicate Number of organisms/chamber 10 Replicates 3 Endpoints Survival Test temperature 25±2°C
Dilution water UBW Photoperiod Permanent dark Test protocol Standard Oper ating Procedure MicroBioTests Control acceptability criteria <10% mortality

Reference test K 2 Cr 2 O 7

Statistics

EC 50 s were calculated as being the concentration/ dilution that causes 50% mortality, using the formula for a dose-response curve with variable slope built-in in GraphP ad Prism. GraphPad Prism uses the following formula:

Hillslope x EC

Bottom Top Bottom y
* ) 50 (log 10 1

−

+ − + =

Wherein:

x = Concentration or dilution y = Mortality (as %) Top = Maximum mortality (100) Bottom = Minimum mortality (0)

The NOEC was determined as the highest test c oncentration in which the growth rate was not significantly reduced compared to the control (U BW). This was assessed by means of a one-way ANOVA followed by Dunnettís multiple comparison post-test.

4.2.2.3 Solea solea

The protocol is based upon OECD 203 (1992). Freshly fertilized eggs of the sole ( Solea solea ) were obtained from SOLEA/IMARES, IJmuid en, from a batch produced 19 August 2007. The eggs were stored at 12-13°C and s light aeration until use in the tests.

The test was performed in 100 mL glass beak ers, containing approx. 80 mL of the test concentration. Each concentration was replicated 4-fold. To prevent dilution of the test concentrations, first 40 fertilized eggs, or yolk-sac larvae (depending on availability) were transferred to a rinsing beaker c ontaining the test concentration. From there, 10 organisms were transferred to each replicate. The test beakers were covered with perforated microwave foil to prevent evaporation, and put away at 12-13°C with a 16 h light ñ 8 h dark regime. After 4 days exposure, surviving organisms we re counted in each beaker.MEPC 57/2/7

ANNEX Page 34

I:\MEPC\57\2-7.doc With each test series, a reference test was conducted, using hydrogenperoxide (H 2 O 2 ) as toxicant.

Summary of test conditions for the acute fish te st with eggs/larvae of the sole Solea solea

Test organism Solea solea Test organism source Commercial ha tchery SOLEA/IMARES, Netherlands Test organism life stage Egg ñ yolk-sac larvae

Test duration 96 h Test chamber 100 mL glass beakers

Test solution volume 80 mL Number of organisms/chamber 10 Replicates 4 Endpoint Survival Test temperature 12-15 °C

Dilution water UBW

Photoperiod 16 hours light/8 hours dark

Aeration None Test protocol OECD 203 (1992) Control acceptability criteria <10% mortality

Reference test H 2 O 2

Statistics

EC 50 s were calculated as being the concentration/ dilution that causes 50% mortality, using the formula for a dose-response curve with variable slope built-in in GraphP ad Prism. GraphPad Prism uses the following formula:

Hillslope x EC

Bottom Top Bottom y
* ) 50 (log 10 1

−

+ − + =

Wherein:

x = Concentration or dilution y = Mortality (as %) Top = Maximum mortality (100) Bottom = Minimum mortality (0)

The NOEC was determined as the highest test c oncentration in which the growth rate was not significantly reduced compared to the control (U BW). This was assessed by means of a one-way ANOVA followed by Dunnettís multiple comparison post-test.

4.3 Quality assurance

As a control on the viability of the test organism s, reference tests were performed simultaneously

with the ballast water tests. For Skeletonema costatum and Artemia franciscana, potassiumdichromate (K 2 Cr 2 O 7 ) was used as reference toxicant. The results were compared with

standard values: ISO for S. costatum and manufacturer specifica tions of the batches for A. franciscana

.MEPC 57/2/7

ANNEX Page 35

I:\MEPC\57\2-7.doc For the sole early life stage test, no standards were available. Therefore, H 2 O 2 was used as reference toxicant and the results were compared with previous tests with sole larvae conducted at the IMARES laboratory.

4.4 Results

4.4.1 Reference tests

The results of the reference tests are summarized in table 4.2. Skeletonema costatum appears to be rather insensitive, with 72 h EC 50 values two to four times highe r than is indicated in the ISO guidelines. K 2 Cr 2 O 7 is a yellow solution causing some disturbance in the fluorometric measurements. This probably resulted in chloro phyll-a measurements that were too high for the treatments with 3.2 and 5.6 mg/L, as measurements of cell activity did not support the relatively high levels of chlorophyll-a in these. It was the intention to verify the fluorometirc measurements by performing cellcounts after the final measurements. However, the cells of S. costatum tended to form aggregates during the test which made cell counting impossible.

Table 4.2: Results of reference tests as EC50 in mg/L

Test initiation

date S. costatum

EC 50 K 2 Cr 2 O 7 A. franciscana

EC 50 K 2 Cr 2 O 7 S. solea

EC 50 H 2 O 2 Expected EC 50 2.5±1.1 26.7-35.6 84-150

22-08-2007 5.6 28.4 87.4

23-08-2007 9.2 54.0 87.6

24-08-2007 8.7 37.8 145.6

25-08-2007 5.6 46.9 -

The EC 50 values obtained in the reference tests with Artemia franciscana were close to or within the indicated range specified for the batch of cysts used at 22- 08 and 24-08, but were higher for

the other two tests. The latter were initia ted by a less experienced analyst, who probably introduced too much hatching medium into the rinsing wells, causing dilution of the test

solutions.

The EC 50 values for H 2 O 2 in the reference tests with sole eggs and larvae, were within the range established in previous tests with this species (Sneekes et al. , 2007). In the test initiated on 25 August the concentrations were accidentally only 10% of those intended. Therefore, the highest concentration was we ll below no-effect level and effects were not observed.

Ballast water tests

Skeletonema costatum

S. costatum responded directly to exposur e to treated ballast water. Each test day, a clear dose response relationship was obvious at the initial meas urements that were performed within half an hour after the algae were introduced into the te st medium (see table 4.3).MEPC 57/2/7

ANNEX Page 36

I:\MEPC\57\2-7.doc Table 4.3: Measurements of chlorophyll-a (chl-a) and cell activity (% act) at initiation of the tests

Start day 24 h 48 h 72 h 96 h

% TBW chl-a % act chl-a % act chl-a % act chl-a % act

0 20.83 31.53 23.22 30.89 21.50 24.24 29.04 27.75

10 19.69 32.11 20.80 38.16 18.80 26.02 29.79 28.74

18 19.51 32.16 21.69 34.74 14.82 23.88 28.91 28.89

33 19.97 30.79 21.23 33.43 13.74 19.58 28.53 30.13

56 16.38 13.56 21.77 35.43 12.16 10.43 25.98 24.09

100 14.31 12.03 20.87 26.00 11.55 10.37 18.86 11.57

In the tests with 24 h and 48 h ol d ballast water, the algae seemed affected in all ballast water dilutions, but recovered during the test in the most diluted treat ments (Figure 1). Algae in 33% TBW grew a little less than in 10% and 18% TBW, but the growth rate was only significantly
different from UBW in the test with 48 h old ballast water.

In 56% TBW the same pattern was observed, but mo re severe. In the test with 24 h old ballast water, the algae died and chlor ophyll-a concentrations decreased to levels below the detection limit. In the other tests, however, the algae recovered afte r an initial decrease in density, resulting in a complete recovery after 3 days in the test with 96 h old ballast water.

In 100% TBW, the algae died within 24 h exposure and no recovery was observed in any of the

four tests.

0 24 48 72 0 100 200 300 400



### 0% TBW

10% TBW

18% TBW

33% TBW

56% TBW

100% TBW

hoursalgal biomass (μg chl-a / L)

Figure 1: Growth of S. costatum introduced into ba llast water dilutions 24 h after treatmentMEPC 57/2/7

ANNEX Page 37

I:\MEPC\57\2-7.doc From the obtained growth rates, the following EC 50 and NOEC values (as % TBW) were calculated (table 4.4):

Table 4.4: EC 50 values based upon algal growth rate

Time after BW treatment 72 h EC 50 72 h NOEC
24 h 34.8% 32.6%
48 h 59.4% 17.8%
72 h 56.7% 32.6%
96 h 67.6% 56.2%

The first and last test (24 h and 96 h old ballast water) were inc ubated in a temperature-controlled shaking cabinet with fluoro-type lights at 20°C. The second and th ird test were incubated in a temporary location with cool-white lights at ca . 18°C and had to be shaken manually. Under these circumstances, the algae grew slower and di d not reach the minimum growth rate of 0.9/d. However, the response to TBW (and the reference toxicant) was similar in all tests, indicating that the lower growth was not due to a lower vi ability of the algae, but to less than optimal

culture conditions.

Artemia franciscana

The brine shrimp A. franciscana was not affected by treated ba llast water. Survival was at least 90% in all treatments, with two exceptions : UBW in the 48 h test (87%) and 10% TBW in the 72 h test (77%).

Solea solea

Survival of sole larvae was good (87.5% to 100%) throughout all tests. No effects of ballast

water treatment were observed.

4.5 Conclusions

At 24 h after treatment of the ballast water with the electrolytic cell, no effects were observed of the treatment on larvae of the brine shrimp ( Artemia franciscana ) and eggs and yolk-sac larvae of the sole ( Solea solea ).

The diatom Skeletonema costatum was severely affected by the treated ballast water, resulting in complete mortality of the algae in undiluted treated ballast water even in the tests initiated 96 h after treatment. In time, however , the effects were gradually alleviated, resulting in complete recovery of the algae in 56% TB W during the 3 day exposure in th e test initiate d with ballast

water 96 h after treatment.MEPC 57/2/7

ANNEX Page 38

I:\MEPC\57\2-7.doc 5 RISK ASSESSMENT (THE M ETHODOLOGY, SECTION 7)

5.1 Risk to safety of the ship

Corrosion

The electrolytic cell itself, including connecting pipes, is fully protected by special coatings against corrosion. The ballast wa ter tanks in principle are subj ect to corrosion, especially immediately after treatment. FAC, however, disappears rapidly. Ballast water tanks normally are coated against corrosion. This regular practice is expected to be sufficient for treated ballast water. Corrosion is at present monitored and will be reported in the application for Final Approval.

Generation of hydrogen gas

During electrolysis hydrogen gas (H 2 ) is produced (see chapter 2). Theoretically, the cell produces 0.0186 mol H 2 gas per ampere (A) per hour, which equals 7.5 g H 2 per hour under standard operating with 200 A. With a flow in the cell of 100 m 3 /hour, this results in a H 2 gas concentration of less then 0,1 ppm. This is far below any dangerous concentration. During full-scale testing for the Fi nal Approval application, H 2 will be monitored.

Other issues

Storage and handling of substances is not an issue, since FAC is produced in situ. Neither is contact with, inhalation or noise an issue. The BWMS will operate in th e machine room of the ships using standard pumps. Neither the Sediment or nor the electrolytic cell produces any noise.

5.2 Risks to human health

FAC

FAC was detected in treated ballast water at a maximum concentration of 1.67 mg/l immediately

after treatment. It decreased to 0.5 mg/l within 2 hours. Ref. 31 states that, in general, regulations

for FAC concentration in swim ming pools require a minimum of 0.4 mg/l. Sometimes maximum concentrations are set, e.g., in France. The figure is 1.4 mg/l. WHO (2004) recommends a drinking water standard for ch lorine of 5 mg/l. The PEC in a standard OECD-EU harbour for

FAC was calculated at 0.34 μ g/l with MAM-PEC (see chapter 3.1). It is concluded that FAC in

treated ballast water in the tanks nor in th e environment poses a risk to human health.

Tribromomethane

Tribromomethane was detected in treated balla st water at a maximum concentration of 290 μ g/l. WHO (2004) recommends a drinking wate r standard for trib romomethane of 100 μ g/l. The PEC
in a standard OECD-EU harbour for tr ibromomethane was calculated at 1.28 μ g/l with MAM-PEC (see chapter 3.2). It is c oncluded that tribromomethane in treated ballast water in the tanks nor in the environment pos es a risk to human health.MEPC 57/2/7

ANNEX Page 39

I:\MEPC\57\2-7.doc 2,2 Dibromoacetic acid

The 2,2 Dibromoacetic acid was detected in trea ted ballast water at a maximum concentration

of 64 μ g/l. It was below detection (1 μ g/l) after 24 hours aging of the ballast water. WHO (2004) recommends a drinking water standard for 2,2 dibromoacetic acid of 100 μ g/l. The PEC in a standard OECD-EU harbour for 2,2 dibr omoacetic acid was calculated at 0.019 μ g/l with MAM-PEC (see chapter 3.3). It is concluded that 2,2 dibromoacetic acid in treated ballast water

in the tanks nor in the environmen t poses a risk to human health.

5.3 Risks to the aquatic environment

FAC

The MAM-PEC model calculates for FAC a steady state situation (= PE C) in the harbour

of 0.34 μ g/l and of 0.017 μ g/l in the surroundings. The model calcu lates a steady state situation in the shipping lane of 0.014 μ g/l.

In various documents, the predicted no effect concentrations of FAC (PNEC) have been
calculated with different outcomes. Annex 7 to MEPC 56/2/2 (review by the GESAMP-BWWG of the application by Japan, ME PC56/2), reports a value of 0.07 μ g/l. Reference 31 reports a

value of 0.04 μ g/l. MEPC 57/2/1 (application by Korea) reports 8 μ g/l.

When using the lowest figure of 0.04 μ g/l the PEC/PNEC ratio for a standard OECD-EU harbour

is 0.34/0.04 = 8.5 and for its surroundi ngs 0.017/0.04 = 0.42. For th e shipping lane it is 0.014/0.04 = 0.35. The applicant concludes, that using these input data , the PEC/PNEC ratio

for the harbour is > 1. In the othe r compartments it is lower than 1.

Tribromomethane

The MAM-PEC model calculates for tribromomethan e a steady state situation (= PEC) in the

harbour of 1.28 μ g/l and of 0.036 μ g/l in the surroundings. The m odel calculates a steady state situation in the shipping lane of 0.005 μ g/l.

In various documents, the predicted no effect c oncentrations of tribromomethane (PNEC) have

been calculated with different outcomes. Annex 7 to MEPC 56/2/2 (review by the GESAMP-BWWG of the application by Japa n, MEPC 56/2), repo rts a value of 7.1 μ g/l. Reference 31 reports a value of 146 μ g/l. RIVM (2005, ref. 30) reports a PNEC of 1.5 μ g/l.

When using the lowest figure of 1.5 μ g/l, the PEC/PNEC ratio for a standard OECD-EU harbour is 1.28/1.5 = 0.85 and for its surroundings 0.036/1.5 = 0.024. For the sh ipping lane it is 0.005/1.5 = 0.003. The applicant concludes, that the PEC/PNEC ratio for tribromomethane is much less than 1 for all compartments.

2,2 Dibromoacetic acid

The MAM-PEC model calculates for 2,2 dibromoacetic acid a steady state situation (= PEC) in

the harbour of 0.019 μ g/l and of 0.001 μ g/l in the surroundings. The model calculates a steady

state situation in th e shipping lane of 0.0007 μ g/l.MEPC 57/2/7

ANNEX Page 40

I:\MEPC\57\2-7.doc In various documents, the predicted no effect c oncentrations of 2,2 dibromoacetic acid (PNEC)

have been derived with different outcome s. Annex 7 to MEPC 56/2/2 (review by the GESAMP-BWWG of the application by Japa n, MEPC 56/2), repo rts a value of 6.9 μ g/l. Reference 31 reports a value of 0.72 μ g/l.

When using the lowest figure of 0.72 μ g/l, the PEC/PNEC ratio for a standard OECD-EU

harbour is 0.019/0.72 = 0.03 and for its surroundings 0.001/0.72 = 0. 001. For the shipping lane it is 0.0007/0.72 = 0.001. The applicant conclu des, that the PEC/PNEC ratio for 2,2 dibromoacetic acid is much less than 1 for all compartments.

5.4 Conclusion

The main concern of this BWMS is the risk to algae by the PEC of FAC in a harbour of limited dimensions (0.34 μ g/l ) and the tribromomethane concen tration (approximately 0.300 mg/l) for the crew in the ballast water tanks after intake. The toxicity for algae was confirmed by the ecotoxicological tests commissioned by the applicant (see chapter 4) . The applicant will further investigate and assess these risks during full scale land-based tests for the Final Approval application.

6 ASSESSMENT REPORT (THE METHODOLOGY, SECTION 8)

6.1 Overview of data and endpoints

Chapter 5 presents an overview of all critical da ta and endpoints used fo r the risk assessment.

All data is science based a nd literature references are given. Monographs on individual compounds have been used among which a draft RAR for the Active Substance. The applicant has commissioned research on the Active Substance and Relevant Chemicals by accredited commercial laboratories and University instituti ons in the Netherlands and Germany. Quality control protocols were at all tim es strictly followed: see annexe s 1-4. Representatives of Lloydís register have consequently witnessed critical tests in the laboratory and on land and have approved these. The applicant considers the qu ality of test reports ìstate of the artî.

6.2 Assessment of risks, input da ta for modeling and uncertainty

From chapter 5 it can be concluded, that the main risk of this BWMS is the PEC of FAC for algae in the harbour of discharge of treated ba llast water. The PEC/PNEC ratio is clearly too high, when calculated with MAM-PEC and the input data used. Some of these input data has a considerable uncertainty:

The volume of discharge per day of ballast water : At present no reliable data is available. Figures are used that differ or ders of magnitudes (50,000 ñ 397,000 m 3 ). The figure used in this application (200,000 m 3 ) is considered too high, according to information of the Rotterdam Port Authority. It implies that each day 3-4 big bulk carriers and/or oil tankers with this BWMS discharge in the same harbour. A more realistic figure is 5 such ships each week. It should be kept in mind that MAM-PEC is a ìsteady stateî m odel, that assumes that there is a permanent source of a pollutant, in this case 334 kg hypochlorit e each day in the same place, the year round. It is highly questionable, whether this is a re alistic assumption. It prob ably is a considerable

overestimation.MEPC 57/2/7

ANNEX Page 41

I:\MEPC\57\2-7.doc Different half-lives : In the literature, vary ing values for the half- life of hypochlorite can be

found: minimum 45 minutes and ma ximum 90 minutes. This is the half-life based on hydrolysis. FAC, however, reacts very quickly with organic material and will disappear very fast in an environment with organic material (see annex 2 in re f. 31). It is difficult to quantify this figure,

but even in the ballast ta nk it reduces from 1.67 to 0.5 mg/l in 2 hours. Further analysis will be done during full-scale land-based tests.

Different concentrations: The T=0 concentration during the tests for this application was 1.67 mg/l. The applicant will optimize th e electrolytic cell to produce the minimum

concentration of FAC for effective ki lling the organisms, which is 0.5 mg/l.

Different PNECS: A critical value for risk assessment is the PNEC value. In this application, the lowest value found in literature (0.04 μ g/l) has been used to calculate the PEC/PNEC ratio.

Further research will be done to decide on the most realistic value of PNEC for hypochlorite, including temperature effects.

6.3 Compliance with the Convention

The results of the laboratory an d land-based tests clearly demons trate that the applicant is complying with the Convention. The BWMS is achieving the minimum killing rates required for all relevant organisms easily through the comb ination of sediment removal and subsequent disinfecting.

Preliminary ship-board tests have shown that the BWMS is effec tive in water with a salt content of > 3 PSU. So, the BWMS can op erate in 96% of all situati ons worldwide. The BWMS can operate in fresh water conditions, but salt has to be added during ballast water intake to a
minimum concentration of 3 PSU.

6.4 Risk management

The applicant considers risks under normal operat ional conditions of this BWMS within the limits of the normal daily operation of any ship. Th e BWMS is a very robust device with little

risk of failure. When switche d of, e.g., under SOLAS, the BWMS immediately stops producing

Active Substance.

The concentration of the Active Substance and Relevant Chemicals will be controlled through the functioning of the electrolytic cell. The BWMS is controlled on line through sensors for flow, pressure and electrical conductivity , which has direct relation with chloride concentration in the ballast water. During full- scale land-based testing, optimization will be inve stigated in order to minimize the production of FAC and Relevant Ch emicals. During these tests FAC, Relevant Chemicals, H2 and other required parameters will be monitored (TSS, DOC, POC, pH, salinity and temperature). Bromate analysis will be incl uded in the program. Corrosion in ballast water tanks is monitored at present in detail and results will be reported in the application for Final Approval. Reliable statistics on the averag e daily discharge of tr eated ballast water in harbours will be collected to estimate more preci se the input data for MA M-PEC to calculate the PEC. Further research will be done to clarify exis ting discrepancies with regard to the value of the PNEC, in particular for hypochlorite. New ec o-toxicological tests wi ll be done with treated ballast water, starting these tests immediately afte r treatment (T=0) and after 5 days of aging. All tests will be done under supe rvision of Lloydís Register.MEPC 57/2/7

ANNEX Page 42

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