I:\MEPC\61\2-12.doc 

# E

MARINE ENVIRONMENT PROTECTION

COMMITTEE

61st session Agenda item 2 MEPC 61/2/12

31 March 2010 Original: ENGLISH

HARMFUL AQUATIC ORGANISMS IN BALLAST WATER

Application for Basic Approval of the BlueSeas Ballast Water Management System

Submitted by Singapore

SUMMARY

Executive summary: This document contains the non-confidential information for Basic Approval of the BlueSeas Ballast Water Management System in accordance with the "Procedure for approval of ballast water management systems that make use of Active Substances (G9)" adopted by resolution MEPC.169(57). The relevant documents and application dossier will be provided to the Marine Environment Division of the Organization for review by the GESAMP-Ballast Water Working Group.

Strategic direction: 7.1

High-level action: 7.1.2

Planned output: 7.1.2.5

Action to be taken: Paragraph 6

Related documents: BWM.2/Circ.13 and BWM.2/Circ.24

Introduction

1 Regulation D-3.2 of the International Convention for the Control and Management of ships' Ballast Water and Sediments stipulates that ballast water management systems which make use of Active Substances to comply with the Convention shall be approved by the Organization, based on a procedure developed by the Organization.

2 The "Procedure for approval of ballast water management systems that make use of Active Substances (G9)" was adopted by reso lution MEPC.169(57). Procedure (G9) defines the principal aspects to be documented by data or testing (MEPC 57/21, annex 1, paragraph 4.2.1) and some basic principles for risk evaluation (MEPC 57/21, annex 1, paragraph 5.3). According to section 6 of Procedure (G9), the Organization should evaluate the information provided in the application.MEPC 61/2/12 Page 2

I:\MEPC\61\2-12.doc 3 According to BWM.2/Circ.13 and BWM.2/Circ.24, proposals for approval of ballast water management systems that make use of Active Substances need to be submitted to the Marine Environment Division of the Organization to be reviewed by the GESAMP-BWWG.

4 The competent authority in Singapore has verified the application dossier prepared by Envirotech and Consultancy Pte. Ltd., and believes that it satisfies the data requirements of Procedure (G9) adopted by resolution MEPC.169(57).

5 Singapore therefore submits the non-confidential part of the applicant's dossier in the annex of this document to the Organization for evaluation according to Procedure (G9). The complete dossier will be made available to the experts of the GESAMP-BWWG.

Action requested of the Committee

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

***MEPC 61/2/12 Annex, page 1

I:\MEPC\61\2-12.doc ANNEX

NON

- CON F IDENTIAL I NFO R M AT I O N O N TH E

BLUESEAS BALLAST WATER MANAGEMENT SYSTEM

Contents

1 INTRODUCTION

1.1 Overview 1.2 System (G9: 7) 1.3 Working mechanism 1.4 Purpose of application

2 IDENTIFICATION (G9: 4.1)

2.1 Active Substances (G9: 2.1.1) 2.2 Preparations (G9: 2.1.3) 2.3 Relevant Chemicals (G9: 2.1.4)

3 DATA-SET FOR SUBSTANCES (G9: 4.2, 5.3)

3.1 Free active chlorine (G9: 4.2.1) 3.2 Hypobromous acid/Hypobromite ion (G9: 4.2.1) 3.3 Hydroxyl radical (G9: 4.2.1) 3.4 Ozone (G9: 4.2.1)

3.5 Sodium thiosulfate

4 TOXICITY ANALYSIS ON DISINFECTION BY-PRODUCTS (G9: 5.2)

4.1 Trihalomethane (THM) 4.2 Haloacetic acids (HAA)

4.3 Bromate

5 TOXICITY TESTING OF THE TREATED BALLAST WATER (G9: 5.2)

5.1 pH

5.2 Total residual oxidant 5.3 Disinfection by-products 5.4 Eco-toxicity testing

6 RISK ASSESSMENT

6.1 Safety of ship (G9: 6.3) 6.2 Human and environment (G9: 6.4)

7 ASSESSMENT REPORT (G9: 4.3, 4.2.3)

7.1 Experimental data 7.2 Literatures for toxicity analysis

REFERENCESMEPC 61/2/12 Annex, page 2

I:\MEPC\61\2-12.doc 1 INTRODUCTION

1.1 Overview

Ballast water is an essential component for the stability of vessels during the voyage. During the ballasting process, living organisms enter the ballast tanks together with ballast water and sediments. Some of these organisms are able to survive during the journey and may be able to grow in the receiving environment upon deballasting. The ballast water mediated invasions have been observed and reported. Some of these invaders can greatly affect the native living organisms and thus the local economic activities, such as fishing, recreation and hydro-power generation. Moreover, there is a ri sk of spreading diseases such as "cholera" through ballast water (Hallegraeff and Bolch., 1991). Inappropriate ballast water management system (BWMS) may lead to en vironmental pollution, economical loss and spread of diseases in our coastal areas.

The ballast water exchange at mid ocean has been widely practiced. Ballast water collected from a coastal area is exchanged with deep seawater during the voyage. It is hypothesized that the organisms which are in the deep seawater will not be easily adapted to the coastal environment. In addition, the organisms in coas tal seawater may not be able to survive in the low-nutrient environment in deep seawater. However, the efficiency of ballast water exchange in removing living organisms is questionable (Taylor et al ., 2007). Moreover, the operation of ballast water exchange is unsafe to the ship structure and the crew. Therefore,

the need for a safe and cost-effective alter native treatment technology is increasing.

1.2 System (G9: 7)

BlueSeas Ballast Water Management System of Envirotech and Consultancy Pte. Ltd., Singapore, has been developed and tested since 2007. The system is designed for treatment of ballast water (seawater) in order to better prevent the marine environment. The BWMS consists of three key components as follows:

.1 a micro-sized strainer (50 μm) in order to remove bigger organisms and particles before entering the electrochemical reactor / ballast water tank;

.2 an electrochemical reactor (also called as electrochemical disinfector or electro-disinfection reactor) for generation of disinfectant (total residual oxidant (TRO) less than 12 mg/L), which are used in the inactivation of organisms in the ballast water. The modular design can be scaled-up according to ballast water capacity; and

.3 a neutralization system to remove residual chlorine prior to its discharge to

ensure that the water is safe to the marine environment.

In addition to the above three components, the BWMS has supporting devices; they include sensors and transmitters (TRO, and optional: flow rate, conductivity, pressure and temperature), computer contro l system, and auxiliary devices.

At the electrochemical reactor, Active Substances including free active chlorine and hydroxyl radical (OH ï ) are produced. The TRO (mainly hypochlorite and hypochlorous acid) with concentration of less than 12 mg/L is generated in reactor; the concentration is measured by an online analyzer, which provides information for the operation of the electrolysis device controlled through a computer system.MEPC 61/2/12 Annex, page 3

I:\MEPC\61\2-12.doc Hydrogen gas (H 2 ) and chlorine gas (Cl 2 ) are generated. A ventilation system is used to remove the gases to prevent fire/explosion and spread of toxicant to crew (e.g., inhalation); it is optional as concentrations of both gaseous species are with extremely low concentration levels (confidential supporting information "SI-list of experiment details").

The system diagrams for operations in both ba llasting and deballasting are given in the confidential supporting information "SI-Description for the BWMS".

Operation of system during ballasting

During the ballasting, the seawater is first pumped through the strainer, goes through the electrochemical reactor, and finally flows to the ballast water tank.

Operation of system during deballasting

The ballast water from each tank is pumped out through a pipe, in which neutralizing agent (Na 2 S 2 O 3 ) is injected. This is to ensure that the water will have no Active Substances

released to the sea and thus have no adverse effects on the environment. The details in the application and handling of the chemical are addressed in the confidential supporting information "SI-application of Na 2 S 2 O 3 ".

The electro-disinfection system in this applic ation treats the safety of the receiving environment, the ship, and the crew as the top priority. No hazardous chemical is needed to be handled by the crew or stored. In addition, the lower foot-print design ensures ease in integration of the system in newly built and existing ships. Lower operation and maintenance costs make the system more affordable than many commercially available technologies.

1.3 Working mechanism

Electrochemical generation of Active Substanc es is the major working mechanism in the ballast water treatment technology in this application. HOCl/OCl
- and hydroxyl radical are two major Active Substances that are produced in the reactor.

1.3.1 Total residual oxidants

1.3.1.1 Free active chlorine

The combination of HOCl, OCl
- and Cl 2 (aq) is referred to as free active chlorine. Free active chlorine is generated due to the production of chlorine gas at the anodes of the

electrochemical reactor as a result of oxidation of Cl
- ions. See below:

2 

### 2 2 ()

Cl e Cl g

− −



### − → (1-1)

The produced chlorine gas immediately dissolves and reacts with the water to form hypochlorous acid (HOCl), which is a weak acid. The dissociation of HOCl forms the hypochlorite ion (OCl
- ). The reactions are as follows (Metcalf and Eddy, 2003;

Chen et al ., 2005):

2 2 

### Cl H O HOCl Cl H − + 

### + ⇔ ++ K H = 4.5x10 -4 M 2 , T= 25 o C (1-2)

H

OCl ClO H − + ⇔ + K i = 3x10 -8 M, T= 25 o C (1-3)

Because of the magnitude of reaction constant of Equation (1-2) shown above is high, large quantities of chlorine can be dissolved in water.MEPC 61/2/12 Annex, page 4

I:\MEPC\61\2-12.doc Figure 1-1: Species distribution of active chlorine at different pH values

Figure 1-1 shows the speciation of free active chlorine at different pH values (Deborde and von Gunten, 2008). At pH > 4, the aqueous-phase chlorine does not exist (White, 1992). In the ballast water, the pH of seawater is around 8 (7.5 to 8.5). Thus, no Cl 2 (aq) exists. Approximately 80% of active chlorine is OCl - and 20% is HOCl.

Free active chlorine is a very powerful disinfectant due to its higher oxidation capability. It stays in water (as residual chlorine) for a period of time ranging from several hours to 1-2 days dependent upon the characteristics of seawater. Hence, a variety of living organisms and micro-organisms originally existing in the ballast water intake (seawater from contaminated coastal area) are disinfected. In addition, the organisms/micro-organisms originally existing in the ballast water tank will be killed and the reactivation of disinfected organisms/micro-organisms does not occur after disinfection because of the presence of the free active chlorine. On the other hand, free active chlorine decays with time, leaving no hazardous concentration after 1-2 days from the point of generation to a lower level (almost zero when the initial chlorine concentration is 3-4 mg/L) (please refer to the confidential supporting information "SI-Chlorine decay in the ballast water"). Therefore, the corrosion due to using the free active chlorine in this technology should not be concerned.

1.3.1.2 Hypobromous acid/Hypobromite ion

It should be noted that, normally, natural seawater contains bromide (Br
- ) ions with a concentration of 35 to 70 mg/L. The produced free active chlorine in seawater will oxidize bromide ions into hypobromous acid according to the following equation:

HOCl + Br
- → HOBr + Cl
- (1-4)

Hypobromous acid can also dissociate in water depending upon pH, temperature and ionic strength, which is similar to hypochlorous acid. Hence, trace of HOBr and OBr
- may exist in

the treated seawater.

HOBr → H + + OBr
- (1-5)

Hypobromous acid is also an effective disinfectant.

1.3.1.3 Total residual oxidants

Chlorine may react with ammonia (NH 3 ) to form chloramines (NH 2 Cl, NHCl 2 and NCl 3 ). Similarly, bromamines can be formed according to the reactions between HOBr and NH 3

.MEPC 61/2/12 Annex, page 5

I:\MEPC\61\2-12.doc The chloramines and bromamines can also act as disinfectants. The overall of free active chlorine, HOBr/OBr
- , chloramines and bromamines are called as total residual oxidants (TRO). In the ballast water treatment in this application, due to the high concentration of Cl
and lower concentration of ammonium and Br
- in seawater, the TRO mainly results from the free active chlorine (i.e. TRO ≈ [HOCl] + [OCl
- ]).

1.3.2 Hydroxyl radical

In the electro-disinfection system in this application, the electro-chlorination is the most important mechanism. However, the effect of hydroxyl radical (OHï) generated at the anode also plays a certain role in the killing of organisms/micro-organisms. Equation (1-6) shows the reaction in the generation of OH as follows:

2 

### H O OH e H − + 

### → ⋅+ + (1-6)

OHï is a strong oxidant and therefore can di sinfect living organisms/micro-organisms. However, the lifespan of OHï is in the scale of nanoseconds, which results in no residuals for further disinfection in the ballast tanks. As a result, the disinfection due to OHï has only

occurred in the electro-disinfection reactor.

1.3.3 Ozone and oxygen

Apart from TRO and OH, there is a possibility of generation of O 2 and O 3 at the anode. The following equations show the reactions at anodes on the generation of oxygen and ozone:

2H 2 O = O 2 (g) + 4H + + 4e - (1-7)

O 2 (g) + H 2 O = O 3 (g) + 2H + + 2e - (1-8)

Because of the higher overpotential of anode (used in the technology in this application), the oxygen generation is limited. The ozone generation is therefore expected to be very small according to the reaction. In addition, our laboratory analysis revealed that the concentration of ozone in the effluent from the reactor was negligible ("SI-list of experiment details").

1.3.4 Others

Besides, such factors as direct electrical field (electric shock) may disinfect a certain portion of living organisms/micro-organisms, without involv ing any Active Substance. However, this is not considered a major mechanism of disinfection because of the low strength of electrical

field used in the reactor.

Sodium thiosulfate is used to remove TRO from the ballast water prior to the discharge. The

main reaction involved in the neutralization is as follows:

Na 2 S 2 O 3 + 4OCl
- + H 2 O = 2NaCl + 2SO 4 2- + 2HCl (1-9)

Na 2 S 2 O 3 + 4OBr
- + H 2 O = 2NaCl + 2SO 4 2- + 2HBr (1-10)

The toxicity of sodium thiosulfate at lower concentration in water solution is insignificant as demonstrated in section 3. As the dosage needed is quite low as stated in the confidential supporting information "SI-Description for the BWMS", its effect on marine environment may be neglected. It can consume dissolved oxygen (DO) according to the below reaction:

2Na 2 S 2 O 3 + O 2 = 2Na 2 SO 4 + 2S (1-11)MEPC 61/2/12 Annex, page 6

I:\MEPC\61\2-12.doc As such, a slightly higher dosage of sodium thiosulfate will be added so that the chlorine can fully be neutralized; the left sodium thiosulfate can be removed by DO in the seawater. The confidential support information "SI-application of Na 2 S 2 O 3 " is included in this application on

various issues associated with the chemical.

1.4 Purpose of application

Since our electro-disinfection technology makes use of Active Substances in the ballast water treatment, we are seeking for the Basic A pproval of Active Substances according to Procedure (G9) (resolution MEPC.169(57), docum ent MEPC 57/21, annex 1, "Procedures for approval of ballast water management systems that make use of Active Substances"). This document is prepared to provide the non-confidential information on safety and environmental related issues of our electro-disinfection technology according to Procedure (G9). The complete application dossier inclusive of this document together with two confidential documents ("Laboratory reports" and "Supporting informati on") is submitted to the GESAMP-BWWG for the evaluation of technology for Basic Approval.

2 IDENTIFICATION (G9: 4.1)

2.1 Active Substances (G9: 2.1.1)

The electro-disinfection treatment technology in this application produces Active Substances on board. No chemical is added to produce any other Active Substances. The following substances are identified as Active Substances according to the definitions in Procedure (G9).

2.1.1 Total residual oxidants

The maximum TRO concentration generated on board is less than 12 mg/L. TRO is divided into two major groups: free active chlorine and hypobromous acid/hypobromite ion.

2.1.1.1 Free active chlorine

In the BWMS, the free active chlorine with a maximum allowable concentration of 12 mg/L (i.e. TRO of 12 mg/L as Cl 2 ) is to ensure the complete disinfection of living organisms/ micro-organisms in ballast water. This is particularly important as it can further ensure complete killing of the sediment-bound living organisms/micr o-organisms, which originally exist in the ballast tanks previously containing ballast water for many years. In addition, the residual free active chlorine can disinfect the "tougher" living organisms and prevents re-growth of disinfected organisms. The concentration of free active chlorine drops to a very low level in a period of time (few days) when the ballast water intake is from a contaminated coastal area.

2.1.1.2 Hypobromous acid/Hypobromite ion

As discussed in section 1.3.1.2, Hypobr omous acid/hypobromite ion (HOBr/OBr
- ) can be generated on board. HOBr/OBr
- is a disinfectant and thus is c onsidered as an Active Substance.

2.1.2 Hydroxyl radical

Another Active Substance is hydroxyl radical (OHï). The process of OHï generation is previously explained. Due to the extremely s hort life span, it is nearly impossible to quantify the amount of OHï generated. However, the detection methods such as radical scavengers show the presence of OHï in the electro-disinfection reactor. OHï contributes to the disinfection of living organisms/micro-organisms in the ballast water. As OHï exists only for nanoseconds, the residual disinfection effects are not expected from this Active Substance.MEPC 61/2/12 Annex, page 7

I:\MEPC\61\2-12.doc 2.1.3 Ozone

Ozone is another Active Substance which ma y be generated inside the electrochemical reactor (concentration is insignificant). Due to the fast decomposition, ozone does not leave any residual disinfectant effect at the ballast tank.

2.2 Preparations (G9: 2.1.3)

According to Procedure (G9), paragraph 2.1.3, no preparations for Active Substances used in ballast water management are identified in the electro-disinfection technology in this application.

2.3 Relevant Chemicals (G9: 2.1.4)

Two major classes of Relevant Chemicals are identified due to their importance.

2.3.1 Hydrogen gas

Hydrogen gas (H 2 ) is produced at the cathodes of the electro-disinfector. The following equation shows the reaction responsible for production of H 2 :

2 2 1

2 H O e OH H

− − +→ + (2-2)

H 2 is known as an explosive gas. The lowe r and upper flammability limit values of H 2 gas are reportedly 4 and 75%, respectively. The content of H 2 generated is low according to our calculation based on the energy consumption and generation of chlorine. Moreover, the hydrogen gas quickly escapes from the tank due to its very small density (its molecular weight is 2). Hence, the concentration of hydrogen gas accumulated in the ballast tank can be neglected. The quantification of H 2 gas produced during the operation is provided in the confidential supporting information "SI-Calculation of hydro gen concentration". A gas ventilation system is optionally incorporated in the BWMS in this application.

2.3.2 Disinfection by-products

Disinfection by-products (DBPs) may be produced during the disinfection as a result of reactions between the disinfectants and the natural organic matter or inorganic substances in ballast water. Trihalomethanes (THM), haloacetic acids (HAAs), and bromate (BrO 3 - ) are identified as the major DBPs produced during the operation of BWMS. The quantification of these DBPs in treated water is provided in a latter part of this report.

3 DATA-SET FOR SUBSTANCES (G9: 4.2, 5.3)

3.1 Free active chlorine (G9: 4.2.1)

Free active chlorine is considered as an Acti ve Substance according to the definition in Procedure (G9). It is generated by the elec trolysis reaction previously discussed.

3.1.1 Effects on aquatic plants, invertebrates, fish, and other biota, including sensitive and representative organisms (G9: 4.2.1.1)

It consists of HOCl and OCl
- . Therefore, the following details are collected from the literatures based on studies conducted on the toxicity of HOCl/OCl
- or NaOCl.MEPC 61/2/12 Annex, page 8

I:\MEPC\61\2-12.doc Chlorine gas is first produced through the electrolysis reaction shown early. It is well reported in various literatures that chlorine gas is quickly dissolved once it is generated. In order to confirm this, an experiment was conducted as follows, under the observation of American Bureau of Shipping (Singapore) (ABS).

A gas sample was collected from a simulated ballast tank, in which the ballast water was treated by the proposed BWMS under laboratory conditions. The concentrations of chlorine of the gas samples were determined by a commercial lab. The concentration of chlorine is less than 0.1% (please refer to the confidential supporting information "SI-list of experiment details").

3.1.1.1 Acute aquatic toxicity

The data based on the literature study related to the acute aquatic toxicity of free active chlorine are shown in Table 3-1. Since the technology in this application does not release any treated ballast water without neutralization (removal of residual free active chlorine), acute aquatic toxicity due to free active chlorine in discharging ballast water is unlikely to occur.

3.1.1.2 Chronic aquatic toxicity

The data based on the literature study related to the chronic aquatic toxicity of free active chlorine are shown in Table 3-2. As above-mentioned, since the BWMS in this application does not discharge any treated ballast water without neutralization (removal of residual free active chlorine), chronic aquatic toxicity due to free active chlorine in discharging ballast water is not possible to take place.

3.1.1.3 Endocrine disruption

Sodium hypochlorite is not listed as an endocrine disruptor under any of the following list (Pesticide Action Network (PAN), 2010).

.1 Illinois EPA list;

.2 Keith list;

.3 Colborn list;

.4 Benbrook list;

.5 Danish Inert list; and

.6 EU list.

Based on the above listing, it can be concluded that the free active chlorine may not have any endocrine effects.

3.1.1.4 Sediment toxicity

The organic carbon partition coefficient, K

oc , is a measure of the tendency for a substance to be adsorbed to organic materials in soil, in sediment, or in biological tissues. K

oc and logK

ow of NaOCl are 13.22 L/kg and -3.42 according to EPI Suite by USEPA (EPA, 2008). As the

value of K

oc is quite small (less than 500 L/kg), the free active chlorine will not easily adsorb onto the solids (e.g., soil and sediment) of the receiving environment. Secondly, the active

chlorine exists as OCl
- at pH of 7.5 to 8.5 (seawater or ballast water). Under such a pH, the particles carry negative surface charges. As a result, the adsorption of active chlorine is quite unlikely. It can therefore be concluded that the usage of chlorine will not cause an increase in the toxicity of the sediment.MEPC 61/2/12 Annex, page 9

I:\MEPC\61\2-12.doc 3.1.1.5 Bioavailability/bioaccumulation/bioconcentration

The logP octanol/water (logP

ow ) and bioconcentration factor (BCF) of NaOCl are reported as -3.42 and -2.19 L/kg, respectively. According to section 6.4.1 of Procedure (G9), the bioaccumulation potential needs to be analysed if the logPow is more than or equal to 3 or if BCF > 2000. Thus, it is clear that the potential of bioaccumulation/bioconcentration is negligible for free

active chlorine.

3.1.1.6 Food web/population effects

The concentrations of Active Substances are reduced to a very low level (around 0.01 ñ 0.05 mg/L) due to the neutralization operation. In addition, the water chemistry (e.g., pH of 7.5 to 8.5 in seawater, lower K

ow value, surface charge properties of solids, and

lower K

oc value) is not favourable in the sorption of the free Active Substances onto the solids. No bioaccumulation is expected. As such, it is impossible that they can be adsorbed onto sediment. Thus the adverse effects on the food web and population are negligible.

3.1.2 Data on mammalian toxicity (G9: 4.2.1.2)

3.1.2.1 Acute toxicity

Inhalation

At low pH, if heated, or mixed with acids, NaOCl may release Cl 2 . In case of inhalation, acute toxicity effects such as coughing and choking, burning sensation, laboured breathing, shortness of breath, severe respiratory tract irritation and pulmonary oedema may occur (MSDS, Siemens water technology, 2000). LC 50 of rat through inhalation exposure to NaOCl is 10 mg/L (European Chemicals Bureau (ECB), 2000).

It is worthy to note that the pH of target water to be treated by the proposed system is in the range of 7.5-8.5 under which Cl 2 cannot be formed. In addition, the treated water is not heated or mixed with acids during operation. As a result, inhalation exposure of free active chlorine is not expected in the operation of system in this application.

Oral

Swallowing concentrated hypochlorite solutions may burn the tissues, causes severe abdominal pain, nausea, vomiting, circulatory collapse, confusion, delirium, coma and collapse. Swallowing large quantities can even cause death (MSDS, Horizon Chemical, 2004).

LD 50 of rat/mouse through oral exposure to NaOCl is given in Table 3-4.

Table 3-4 clearly shows that the oral exposure is extremely serious if the concentration of free active chlorine is high. However, the concentration of free active chlorine in the technology in this application is less than 12 mg/L. The substance is generated and exists in the sealed reactor. Thus, no hazardous effect is expected through oral exposure.

3.1.2.2 Effects on skin and eye

Skin contact

Skin contact of 10-12% NaOCl solution may cause irritation, redness, blisters, vesicular eruptions, and eczematoid dermatitis (MSDS, Siemens water technology, 2000). Acute dermal toxicity test on rabbits showed a LC 50 of >10000 mg/kg bw. Severe injury in skin was resulted in 15 to 30 minutes after applying commercial 5.25% NaOCl (diluted 2:1 with water) on rabbits. In addition, a patch test (NaOCl, concentration is not specified) conducted on 225 humans, 3 showed positive reactions (ECB, 2000).MEPC 61/2/12 Annex, page 10

I:\MEPC\61\2-12.doc Eye Contact

If eyes contact with NaOCl, irritation as eyes burn may occur, which is dependent upon the concentration. Eye irritation study using rabbits was reported (ECB, 2000). 5% NaOCl was used in the investigation. Volumes of 0.01, 0.03, and 0.1 ml were contacted with the central corneal surface of the rabbits. Twenty-one days study showed that 0.01 ml resulted in a

moderate irritation while 0.03 ml resulted in a substantial irritation. Severe or corrosive

reactions were observed at 0.1 ml.

The concentrations of free active chlorine in the treated ballast water are quite low and they
are in the sealed devices and ballast water tanks. As such, the risk associated with skin and eye contacts are neglected.

3.1.2.3 Chronic and long-term toxicity

Exon et al. (1987) reported a chronic toxicity study of NaOCl on rats. Rats were given chlorinated drinking water (chlorinated using 6.05% NaOCl) with 5, 15, 30 ppm of available chlorine for 9 weeks. A significantly smaller spleen weights were found among the rats exposed to 30 ppm, compared to the control. However, when consider all levels of dosages, no significant effects of NaOCl exposure we re observed on body weight, thymus weight, antibody response, NKC cytotoxicity, IL2 production, or phagocytic activity.

A two-year NaOCl toxicity study (oral exposure) was reported on rats and mice (U.S. Department of Health and Human Services, 1992). Chlorine with concentrations varied between 0-275 ppm was used. For high dose rats, the dosage was between 13-14 mg/kg. In female rats, an increase in mononuclear cell leukaemia was observed compared to the controls. The survival of rats was not varied significantly either they were exposed or not exposed to chlorinated drinking water. For high does mice, the dosage was between 20-23 mg/kg. Among high dose male mice, a body weight reduction of 5-8% compared to the controlled was observed. Body weight reduction of high dose female mice compared to the controlled was reported as 5-7%. However, the survival rate among the mice was not significantly differed whether they were exposed or not exposed to chlorinated water.

Possibilities of chronic oral exposure of ma mmals are very low in the BWMS, especially because of the neutralization of treated water prior to discharge.

3.1.2.4 Developmental and reproductive toxicity

A series of studies on developmental/reproductiv e toxicity of the Active Substances was reported (U.S. Department of Health and Human Services, 1992). Female Sprague-Dawley rats were exposed to HOCl through drinking water. The concentrations were 0, 1, 10, or 100 mg/L and the duration was 2.5 months prior to conception and throughout gestation. On the 20th day of gestation, the rats were killed. The foetuses were examined for soft tissue and skeletal abnormalities. No increase in foetal resorptions was observed. HOCl exposure in utero produced some soft-tissue defects, such as improper orientation of the heart and adrenal agenesis, if the HOCl concentration was 100 mg/L. A slight increment in skeletal variants, such as incompletely ossified or missing sternebrae or rudimentary ribs was observed (Abdel-Rahman et al., 1982).

Carlton et al. (1986) studied the teratogenic and reproductive toxicity of HOCl (NaOCl). Male Long Evans rats and female rats were investigated at doses of 0, 1, 2 or 5 mg/kg body weight by gavage. Male rats were exposed for 56 days prior to breeding and throughout the 10-day breeding cycle while the female rats were exposed for 14 days prior to breeding and throughout breeding, gestation, and lactation, until the pups were weaned on the 21st day.MEPC 61/2/12 Annex, page 11

I:\MEPC\61\2-12.doc It was reported that no changes in sperm count, sperm motility, or sperm morphology were found or no dose-related effects on fertility, fetal viability, litter size, day of eye opening, and day of vaginal patency were observed. Other studies reported in the literatures on effects of chlorinated drinking water on fetal development in mice found no evidence for a teratologic effect due to the intake of chlorinated drinking water (Chernoff et al., 1979; Kavlock et al., 1979; Staples et al., 1979).

The concentration of free active chlorine in this application is controlled at < 12 mg/L and the chlorine is removed by neutralization prior to the discharge of ballast water. The operations are conducted in sealed reactor and tanks. Living animals and organisms unlikely stay in the ballast water environment. Thus, the developmental and reproductive toxicity can be negligible.

3.1.2.5 Carcinogenicity

HOCl/OCl
- or NaOCl is not listed as a known or probable human carcinogen by the American Cancer Society (accessed on 07-02-2010). However, there are some studies of carcinogenicity on animals (e.g., mice and rats) (e.g., U.S. Department of Health and Human Services, 1992). In the studies by Kurokawa et al. (1986) and Hasegawa et al. (1986), female mice and male rats were exposed (oral) to 1000 mg/L and 200 mg/L of NaOCl, respectively, for 103-104 weeks. No dose-related increment in tumour was found. In addition, a 2-year study conducted by the NTP did not found any evidence of carcinogenic activity of chlorinated water (70-275 mg/L) on male or female mice.

Due to lower concentration of the free Active Substances in the operation in this application and the neutralization, the carcinogenicity is not expected.

3.1.2.6 Mutagenicity

Mutagenic activities of HOCl/OCl
- (NaOCl) on mammals are not well reported in the literatures. Hayashi et al . (1988) conducted a study on mutagenicity of NaOCl by mice. The mice were exposed to NaOCl (6.6%, 312.5-2,500 mg/kg) through a single intraperitoneal injection. No noteworthy increment in the frequency of micronucleated polychromatic erythrocytes in the bone marrow (sampled 24 hours after treatment) of male tidY mice was

identified. Meier et al . (1985) conducted a study on mutagenicity of NaOCl. Chromosome aberrations or micronuclei in CD-1 mice or sperm-head abnormalities in B6C3F1 mice were investigated. The chemical was administered by gavage. Both acute (single dose) and subchronic (5 successive dosings at 24-hour intervals) protocols were employed. It was found that the treatment did not induce micronuclei or aberrations, but did produce a significant dose related increase in sperm-head abnormalities in samples collected 3 weeks after the final subchronic dose. However, no increments were found 1 or 5 weeks after final dosing.

Due to lower concentration of the free Active Substances in the operation in this application and the neutralization, the mutagenicity is not expected.

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

3.1.3.1 Modes of degradation (biotic/abiotic)

Biotic degradation is not expected since it has very high oxidation potential. Abiotic degradation of free active chlorine is possible and the routes are discussed as follows:

.1 The degradation of HOCl/OCl
- (NaOCl) is affected by several factors such as the initial concentration, temperature, pH, light, the presence of other substancesMEPC 61/2/12 Annex, page 12

I:\MEPC\61\2-12.doc (mainly organic contaminants), and physical factors (e.g., mixing/stirring strength). The degradation rate/kinetics is higher at a higher initial concentration. Higher temperature will enhance the rate of degradation.

.2 In addition, the NaOCl is unstable if the pH value is lower than 11. The degradation of NaOCl due to light becomes faster due to photolysis; it becomes much faster in UV light radiation. It is worthy to note that in fresh water or seawater environment, pH of the solution is around 7-8.

Under these conditions most of the free active chlorine exists as OCl
-

. At the maximum sunlight intensity of 330 nm, OCl
- has a molar absorptivity of 15 times greater than HOCl. Thus, the degradation of free active chlorine due to photolysis can be extremely high. Apart from the above factors, the composition of the chlorinated solution affects the rate of degradation. For instant, high organic and inorganic content may increase the rate of degradation due to the oxidation reactions. Degradation of HOCl (NaOCl) may result in a range of by-products which may be harmful depending on the concentration (e.g., trihalomethanes and haloacetic acids).

Chlorine can react with ammonium in water to form NCl 3 , NHCl 2 , and NH 2 Cl, which are also disinfectants. However, the chance of the occurrence is very low as the concentration of ammonium in coastal area is negligible.

.3 The decay kinetics of chlorine in clean and contaminated seawater is given in the confidential supporting information "SI-Chlorine decay in the ballast water". The decay is faster when the seawater is contaminated (like ballast water).

3.1.3.2 Bioaccumulation, partition coefficient, octanol/water coefficient

The logP

ow and BCF of HOCl/OCl
- (NaOCl) are -3.42 and -2.19 L/kg, respectively. As logPow < 3 and BCF < 2000, no bioaccumulation of NOCl is expected. Koc, the organic carbon partition coefficient is a measure of the tendency for a substance to be adsorbed to organic materials in soil, in sediment, or in biological tissues. Koc of HOCl (NaOCl) is 14.3 L/kg. The value of Koc is <500 L/kg and, thus HOCl will not readily partition in to soil/sediment of the receiving environment and no significant sediment to xicity is expected.

3.1.3.3 Persistence and identification of the main metabolites in the relevant media (Ballast water, marine and fresh water)

Due to the rapid degradation, free active chlorine is not considered to be persistent in ballast water, marine or fresh water. Furthermore, due to the low logPow, and BCF values, free active chlorine will not tend to be bioaccumulated in the aquatic life and metabolites.

3.1.3.4 Reaction with organic matter

HOCl/OCl
- (NaOCl) can react with organic matter and produce secondary by-products. Trihalomethanes and haloacetic acids are important by-products as a result of reactions. They may be toxic to various living organisms in the receiving environments. Thus it is important to experimentally determine the conc entrations of the disinfection by-products and study the toxicity of treated ballast water.

3.1.3.5 Potential physical effects on wildlife and benthic habitats

No reports are found in the literatures on the physical effects on wildlife and benthic habitat. However, based on the acute and chronic toxicity related data previously discussed and the toxicity study of our treated ballast water, the physical effects are expected to be negligible because the treated water is neutralized (i.e. remo ve HOCl (NaOCl)) prior to discharge in the operation of the technology in this application.MEPC 61/2/12 Annex, page 13

I:\MEPC\61\2-12.doc 3.1.3.6 Potential residues in seafood

According to the physical and chemical properties of the Active Substances given and discussed above, no residues are expected in seafood due to free active chlorine.

3.1.3.7 Any known interactive effects

No other interactive effects are expected.

3.1.4 Physical/chemical properties of HOCl/OCl - (G9: 4.2.1.4)

Physical/chemical properties of HOCl/OCl
- are given in Table 3-5.

3.1.5 Other details of the Active Substance

Synonyms: Bleach; hypochlorous acid, sodium salt; soda bleach

CAS No.: 7681-52-9 Molecular Weight: 74.44

Chemical Formula: NaOCl

(MSDS, JT Baker, 2009)

3.1.6 Analytical method (G9: 4.2.1.5)

The measurement is conducted according to "Standard Method for the Examination of Water and Wastewater" (APHA, 2005).

3.2 Hypobromous acid/Hypobromite ion (G9: 4.2.1)

3.2.1 Data on effects on aquatic organisms (G9: 4.2.1.1)

3.2.1.1 Acute aquatic toxicity

No data were found for the acute aquatic toxicity of hypobromous acid/hypobromite. However, due to its oxidizing properties, acute aquatic toxicity is expected from these agents.

3.2.1.2 Chronic aquatic toxicity

No information on the chronic aquatic toxici ty of hypobromous acid/hypobromite ion was found in the literature. However, due to its oxidizing properties, chronic aquatic toxicity is expected from these agents.

3.2.1.3 Endocrine disruption

No information on the endocrine disruption of hypobromous acid/hypobromite ion was found in the literature, indicating hypobromous acid /hypobromite ion might not have any endocrine effects on aquatic organisms. Their short half-lives present an extremely low risk.

3.2.1.4 Sediment toxicity

The organic carbon partition coefficient (K

oc ) of hypobromous acid/hypobromite ion is 14.3 L/kg.

As the value of K

oc is quite small (less than 500 L/kg), the hypobromous acid/hypobromite ion will not easily be adsorbed onto the sediment of the receiving environment. As a result, the adsorption of hypobromite ion is quite unlikely. It can therefore be concluded that the hypobromite ion will not cause an increase in the toxicity of the sediment.MEPC 61/2/12 Annex, page 14

I:\MEPC\61\2-12.doc 3.2.1.5 Bioavailability/biomagnifications/bioconcentration

The logP octanol/water (logP

ow ) and bioconcentration factor of hypobromous acid/hypobromite ion are reported as -1.03 and 1.24 L/kg, respectively. According to section 6.4.1 of Procedure (G9) (MEPC 57/21), the bioaccumulation potential is needed to be analysed if the logPow is larger than 3 or if BCF > 2000. Thus, it is clear that the potential of bioaccumulation/bioconcentration is negligible for hypobromous acid/hypobromite ion.

3.2.1.6 Food web/population effects

Hypobromous acid/hypobromite ion is not expected to be bioaccumulated in the food chain, since the value of Log Pow and BCF are very low. On the other hand, their short half-lives present an extremely low risk of such.

3.2.2 Data on mammalian toxicity (G9: 4.2.1.2)

3.2.2.1 Acute mammalian toxicity

No data on the acute mammalian toxicity was found in the literature.

3.2.2.2 Effects on skin and eye

No information on the effects on mammalian's skin and eye was found in the literature.

3.2.2.3 Chronic and long-term toxicity

No information on the chronic and long-term mammalian toxicity of hypobromous acid/hypobromite ion was found in the literature.

3.2.2.4 Developmental and reproductive toxicity

No information on the developmental and reproductive toxicity was found in the literature.

3.2.2.5 Carcinogenicity

No data on the carcinogenicity was found in the literature.

3.2.2.6 Mutagenicity/genotoxicity

No data on the mutagenicity/genotoxicity was found in the literature.

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

3.2.3.1 Modes of degradation (biotic; abiotic)

No data on the modes of degradation (biotic and abiotic) is found in the literature.

3.2.3.2 Bioaccumulation, partition coefficient, octanol/water partition coefficient

The octanol/water partition coefficient (log Pow), bioaccumulation factor, and the Koc for the hypobromous acid/hypobromite ion are -1.03, 1.24, and 14.3, respectively.MEPC 61/2/12 Annex, page 15

I:\MEPC\61\2-12.doc 3.2.3.3 Persistence and identification of the main metabolites in the relevant media (ballast water, marine and fresh waters)

It is unlikely that it can be accumulated through the aquatic organisms and metabolites due

to the low Pow and BCF values.

3.2.3.4 Reaction with organic matter

No available information on reaction with organic matter is available in the literature.

3.2.3.5 Potential physical effects on wildlife and benthic habitats

No potential effects are indicated in the literature.

3.2.3.6 Potential residues in seafood

No potential residues are indicated in the literature.

3.2.3.7 Any known interactive effects

No known interactive effects are indicated in the literature.

3.2.4 Physical and chemical properties for the Active Substances, Relevant Chemicals and treated ballast water (G9: 4.2.1.4)

Physical and chemical properties of hypobromous acid are shown in Table 3-7.

3.2.5 Other details of the Relevant Chemical

Synonyms: hypobromous acid

CAS No.: 13517-11-8 Molecular Weight: 96.911

Chemical Formula: HOBr

http://www.caslab.com/Hypobromous_acid_CAS_13517-11-8/

3.3 Hydroxyl radical (G9: 4.2.1)

Hydroxyl radical is considered as an Active Substance according to the definition in Procedure (G9). It is generated according to the mechanisms previously discussed.

3.3.1 Data on effects on aquatic plants, invertebrates, fish, and other biota, including sensitive and representative organisms (G9: 4.2.1.1)

Toxicity tests carried out to evaluate acute toxicity, chronic toxicity, endocrine disruption, sediment toxicity, bioaccumulation, and food web/population effects on aquatic life due to

OHï are not available in the literatures, due to the extremely short lifespan of the OHï. Although it is highly reactive (oxidation potential of 2.8 V) and able to damage aquatic life at the point of generation, the lifespan of nanoseconds prevents any negative effect of OHï on the aquatic life in discharging environment.MEPC 61/2/12 Annex, page 16

I:\MEPC\61\2-12.doc 3.3.2 Data on Mammalian Toxicity (G9: 4.2.1.2)

The OHï considered as the mediator of the DNA damage due to ionizing radiation (Balasubramanian et al ., 1998; Chen et al ., 2005). In addition, OHï plays an important role in the photolysis of human erythrocytes sensitized by the photosensitization with protoporphyrin IX and ketoporfen (El-Missiry and Abou-Seif, 2000). However, the electro-disinfection technology in this application does not allow any contact between the produced OHï (only inside the reactor) and human/mammalian under any circumstances. Thus, no adverse effect is expected on human/mammalian due to the production of OHï inside the

electro-disinfection reactor.

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

The OHï is known as the "detergent of troposphere" due to its high reactivity with many pollutants. The OHï generated in the BWMS in this application does not reach the discharging environment due to the neutralization operation. Because of the short lifespan of OHï, the bioaccumulation does not occur and the substance is not persistent. The OHï does not cause any effect on the wildlife and benthic habitats or leaves no residues in seafood.

OHï can react with organic and inorganic matters in ballast water. Thus, there is a possibility of generation of DBPs (Xie, 2004; Lui et al ., 2008). It is difficult to differentiate the DPBs produced due to the production of free active chlorine, O 3 or OHï because of the simultaneous generation of free active chlorine and OHï in the proposed reactor system. The analysis of DBPs in the discharging ballast water treated using the technology in this application is presented in a latter part of this application dossier ("SI-list of experimental details").

3.3.4 Physical/chemical properties of OHï (G9: 4.2.1.4)

Physical properties: To the best of our knowledge, not available in literature.

Chemical properties: Hydroxyl radical (OHï) is extremely reactive (oxidation potential of 2.8 V). Thus, it does not exist for more than few collisi ons after it forms in the electro-disinfection reactor. Lifespan of OHï is in the range of nanoseconds. Therefore, the OHï reacts only in the very close proximity to the electrode which it is produced (Bielski and Cabelli, 1995).

3.3.5 Other details of the Active Substance

Synonyms: OH; Hydroxy radical; Hydroxyl

CAS No.: 3352-57-6 Molecular Weight: 17.0073

Chemical Formula: HO

3.3.6 Analytical method (G9: 4.2.1.5)

The measurement is conducted by using radical scavengers such as iso-propanol.

3.4 Ozone (G9: 4.2.1)

3.4.1 Effects on aquatic plants, invertebrates, fish, and other biota, including sensitive and representative organisms (G9: 4.2.1.1)

A zero concentration of ozone was found in ballast water, immediately after the treatment ("SI-list of experiment details"). Ozone decomposes within a very short period of time (in several minutes) and thus, no ozone was found in the discharging ballast water.MEPC 61/2/12 Annex, page 17

I:\MEPC\61\2-12.doc 3.4.1.1 Acute aquatic toxicity

The data related to the acute aquatic toxicity of ozone is shown in Table 3-8. Since the technology proposed in this application does not discharge any ozone to the receiving environment, acute aquatic toxicity due to ozone is highly unlikely to occur.

3.4.1.2 Chronic aquatic toxicity

The data related to chronic aquatic toxicity are not found in literature. The ozone in the treated water is not expected as: 1. the water is neutralized; 2. the decay of ozone is rapid.

3.4.1.3 Endocrine disruption

Ozone is not listed as an endocrine disruptor under any of the following lists (Pesticide Action Network (PAN), 2010): Illinois EPA list, Keith list, Colborn list, Benbrook list, Danish Inert list, and EU list. Thus, ozone may not have any endocrine effects.

3.4.1.4 Sediment toxicity

Due to the fast decomposition of ozone in natural waters, no sediment toxicity is expected. Literature related to sediment toxicity of ozone was not found.

3.4.1.5 Bioavailability/bioaccumulation/bioconcentration

Fast decomposition of ozone in natural waters may not allow any bioaccumulation of ozone.

3.4.1.6 Food web/population effects

The above discussion reveals that ozone does not c oncentrate in organisms or in sediments. Therefore, ozone may not affect the food web.

3.4.2 Data on Mammalian Toxicity (G9: 4.2.1.2)

3.4.2.1 Acute toxicity

Ozone is toxic to the lung. The effects such as pulmonary congestion, and edema can occur if exposed to ozone. Human volunteers exposed at 0.5 ppm, 3 h/d, 6 d/week, for 12 weeks showed changes in lung function (HSDB). However, due to the very low concentrations generate in the proposed technology, no gaseous ozone is expected.

3.4.2.2 Effects on skin and eye

Skin contact

If liquid ozone contacts skin, severe burns may occur (HSDB). The very low concentration which may generate in the current system does not pose such toxicity to the skin.

Eye Contact

Ozone gas may create eye irritation (MSDS- BOC gas es). Such toxicity is not expected due to the very low concentration which may generate in the current system.MEPC 61/2/12 Annex, page 18

I:\MEPC\61\2-12.doc 3.4.2.3 Chronic and long-term toxicity

Immunosuppression occurs in mice and pigs at exposure at low levels of ozone, but not humans. If exposed to ozone concentration > 1 ppm (6 h/d) for 1 year, bronchiolitis and bronchitis in animals is reported (MSDS- BOC gases).

3.4.2.4 Developmental and reproductive toxicity

According to the PAN pesticide database, ozone is not listed as a developmental or reproductive toxin in the following list: CA Prop 65 Developmental toxin; US TRI Developmental toxin, CA Prop 65 Female Reproductive toxin, and CA Prop 65 Male Reproductive toxin. However, ozone is listed as a reproductive toxin under US TRI Reproductive Toxins list.

3.4.2.5 Carcinogenicity

Ozone is not classifiable as a human carcinogen (HSDB).

3.4.2.6 Mutagenicity

Ozone is not reported as a mutagen (MSDS- BOC gases).

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

Ozone which may be generated in the BWMS in this application does not reach the discharging environment. Our experimental testing shows that the concentration is negligible (zero concentration). Ozone is highly reactive and thus may create DBPs in the ballast tank. Bromate is the predominant DBP produced due to ozone. However, the bromate concentration in discharging ballast water was found extremely low ("SI-list of experimental details" and "Laboratory reports"). This suggests that the amount of ozone produced at the reactor can be extremely low as well. Because of the short lifespan of ozone, it does not create bioaccumulation and is not persistent. For the same reason, produced ozone does not cause any effect on the wildlife and benthic habitats or leaves no residues in seafood.

3.4.4 Physical/chemical properties of ozone (G9: 4.2.1.4)

Physical/chemical properties of ozone are illustrated in Table 3-9.

3.4.5 Other details of the Active Substance

Synonyms: Triatomic oxygen

CAS No.: 10028-15-6 Molecular Weight: 48.00

Chemical Formula: O 3

3.4.6 Analytical method (G9: 4.2.1.5)

The measurement is conducted according to "Standard Method for the Examination of Water and Wastewater" (APHA, 2005).

3.5 Sodium thiosulfate

Sodium thiosulfate is used to remove residual oxidant when necessary during the deballasting.

It is not considered as an Active Substance according to the definition in Procedure (G9).MEPC 61/2/12 Annex, page 19

I:\MEPC\61\2-12.doc 3.5.1 Data on effects on aquatic organisms

3.5.1.1 Acute aquatic toxicity

The data related to the acute aquatic toxicity are shown in Table 3-10.

3.5.1.2 Chronic aquatic toxicity

The data related to the chronic aquatic toxicity o are shown in Table 3-11.

3.5.1.3 Endocrine disruption

No information on the endocrine disruption was found in the literature, indicating sodium thiosulfate might not have any endocrine effects on aquatic organisms.

3.5.1.4 Sediment toxicity

No information on the sediment toxicity can be found in the literatures.

3.5.1.5 Bioavailability/biomagnifications/bioconcentration

The logP octanol/water (logP

ow ) and bioconcentration factor of Na 2 S 2 O 3 are reported as -4.53 and 3.162 L/kg, respectively. According to section 6.4.1 of Procedure (G9), the bioaccumulation potential is needed to be analysed if the logPow is more than or equal to3 or if BCF > 2000. Thus, it is clear that the potential of bioaccumulation/bioconcentration is negligible for free active Na 2 S 2 O 3

.

3.5.1.6 Food web/population effects

Sodium thiosulfate is not expected to bioaccumulate in the food chain, since the value of Log Pow and BCF are very low.

3.5.2 Data on mammalian toxicity

3.5.2.1 Acute mammalian toxicity

Oral LD 50 of >5050 mg/kg was reported for rats (EPA Federal Register, Sodium thiosulfate, 2001).

3.5.2.2 Effects on skin, eye, and sensitization effects

Skin: May cause skin irritation Eye: May cause eye irritation and possible burns

Sensitization: May cause respiratory tract irritation.

(MSDS, Sigma-Aldrich, 2009)

3.5.2.3 Repeated-dose toxicity

No data on the repeated-dose toxicity of sodium thiosulfate is found in the literature (MSDS Sigma-Aldrich, 2009).

3.5.2.4 Chronic toxicity

No data on the chronic toxicity of sodium thiosulfate is found in the literature.MEPC 61/2/12 Annex, page 20

I:\MEPC\61\2-12.doc 3.5.2.5 Developmental and reproductive toxicity

Oral exposure did not show any effect on maternal or foetal survival, or in incidences of visceral or skeletal abnormalities in mouse (EPA Federal Register, Sodium thiosulfate, 2001).

3.5.2.6 Carcinogenicity

No data on the carcinogenicity of sodium thiosulfate is found in the literature.

3.5.2.7 Mutagenicity/genotoxicity

Sodium thiosulfate has no mutagenic effects (EPA Federal Register, 2001).

3.5.2.8 Toxicokinetics

Sodium thiosulfate is poorly absorbed from bowel and thiosulfate ion distributes itself in extracellular fluid (HSDB, 2009).

3.5.3 Data on environmental fate and effect under aerobic and anaerobic conditions

3.5.3.1 Modes of degradation (biotic; abiotic)

No data on the modes of degradation (biotic and abiotic) is found in the literature.

3.5.3.2 Bioaccumulation, partition coefficient, octanol/water partition coefficient

The octanol/water partition coefficient (log Pow), bioaccumulation factor, and the Koc for the sodium thiosulfate are -4.53, 3.162, and 6.124, respectively.

3.5.3.3 Persistence and identification of the main metabolites in the relevant media (ballast water, marine and fresh waters)

It is unlikely that it can be accumulated through the aquatic organisms and metabolites due

to the low Pow and BCF values.

3.5.3.4 Reaction with organic matter

No available information on reaction with organic matter is available in the literature.

3.5.3.5 Potential physical effects on wildlife and benthic habitats

No potential effects are indicated in the literature.

3.5.3.6 Potential residues in seafood

No potential residues are indicated in the literature.

3.5.3.7 Any known interactive effects

No known interactive effects are indicated in the literature.

3.5.4 Physical and chemical properties

The physical and chemical properties of sodium thiosulfate can be found in Table 3-12.MEPC 61/2/12 Annex, page 21

I:\MEPC\61\2-12.doc 3.5.5 Other details of the Relevant Chemical

Synonyms: Sodium thiosulfate, pentahydrate; thiosulfuric acid, disodium salt, pentahydrate

CAS No.: 10102-17-7 (Pentahydrate) Molecular Weight: 248.17

Chemical Formula: Na 2 S 2 O 3 .5H 2 O

3.5.6 Analytical method

The concentration is measured by an IC method (IPC-TM-650.2.3.28A).

Table 3-1: Data related to the acute aquatic toxicity (free active chlorine)

Species Explanation Reference

Blackbrow bleak ( Acanthalburnus microlepis De Filippi, 1863) 96 h, LC 50 = 0.6343 mg/L (In fresh water, Using Finney's method and SPSS software) Aksu et al ., 2008

Estuarine fish, Oryzias javanicus 96 h, LC 50 = 0.19 mg/L AÒasco et al ., 2008 Fish

Cyprinus carpio 72 h, NOEC (Genetics) = 50 μg/L PAN pesticide

database

Resting eggs of the

fresh water cladoceran Daphnia mendotae Acute toxicity

LC 90 = 78.3±1.6 mg/L Raikow et al ., 2007

Resting eggs of marine brine shrimp ( Artemia sp. ) Acute toxicity

LC 90 = 86.5±3 mg/L Raikow et al ., 2007

Marine Amphipod, Hyale

barbicornis 96 h, LC 50 = 2.2 mg/L,

EC 50 = 1.1 mg/L AÒasco et al ., 2008

Heptacarpus pictus 20 h, LC 100 = 1600μg/L PAN pesticide

database Invertebrate

Asellus intermedius 96 h, LC 50 = 32000μg/L PAN pesticide

database

Dunaliella primolecta 24 h, LC 50 = 0.4 ppm Videau et al .,1979

Pavlova Iutheri 24h, LC 50 = 3.5-4.0 ppm Videau et al ., 1979

Phaeodactylum

tricornutum 24h, No mortality at 0.6 ppm but growth is stopped/slower down. 0.8 ppm, total mortality Videau et al ., 1979

Green algae (Dunaliella primolecta) LC 50 = 400μg/L PAN pesticide

database Phytoplankton

Chaetoceros decipiens 24 h, EC 50 = 140μg/L PAN pesticide

databaseMEPC 61/2/12 Annex, page 22

I:\MEPC\61\2-12.doc Table 3-2: Data related to the chronic aquatic toxicity (free active chlorine)

Species Explanation Reference

Estuarine fish, Oryzias javanicus

, early life stage Eggs exposed for14 d and the

fish larvae hatched from exposed eggs again exposed for another 21 d, 1 mg/L. No major adverse effects AÒasco et al ., 2008

Cyprinus carpio 30 d, NOEC (Genetics) = 100 μg/L PAN pesticide

database Fish

Menidia beryllina 7 d, NOEC (Growth) = 87μg/L PAN pesticide

database

Marine Amphipod, Hyale barbicornis, early life stage 12 d, 1 mg/L. No major adverse effects AÒasco et al ., 2008

1 mg/L, 100% mortality in 20 d. 7 mm (shell length) 5 mg/L, 100% mortality in 102 h.

1 mg/L, 100% mortality in 24 d. Invertebrates

Brachidontes

striatulus 25 mm (shell length) 5 mg/L, 100% mortality in 156 h. Rajagopal et al

., 1997

Phytoplankton 20 d, about 40-50% reduction in cell density at 0.001-0.25 mg/L Sanders et al ., 1981
Phytoplankton

Porphyra yezoensis 10 d, EC 50 = 1400μg/L PAN pesticide

database

Table 3-3: Criteria for identification of persistent, bioaccumulative and toxic (PBT) substances

Criterion PBT criteria

Persistence Half-life: > 60 days in marine water, or > 40 days in fresh water, or > 180 days in marine sediments, or > 120 days in fresh water sediments

Bioaccumulation Experimentally determined BCF >2000, or if no experimentally BCF has been determined, LogPow ≥ 3

Toxicity Chronic NOEC < 0.01 mg/l

Table 3-4: Values of LD 50 of rat/mouse through oral exposure to NaOCl +

Species LD 50 Remarks

Rat >5000 mg/kg bw ** Liquid bleach containing NaOCl, 4.74% available chlorine

Rat 8200 mg/kg bw NaOCl, 12.5% active chlorine

Mouse 880 mg/kg bw NaOCl

Mouse 5800-6800 mg/kg bw NaOCl, 50%

** bw: body weight.

+ European Chemicals Bureau, 2000.MEPC 61/2/12 Annex, page 23

I:\MEPC\61\2-12.doc Table 3-5: Physical/chemical properties of HOCl/OCl Property Value Reference

Melting point -6°C MSDS, JT Baker, 2009

Boiling point 103.9°C (6% NaOCl) MSDS, Horizon Chemical Co., Inc, 2004

Flammability NFPA rating= 0 MSDS, Siemens water technology, 2000

Relative density 1.2 - 1.25 g/cm 3 MSDS, Sigma-Aldrich, 2009

Vapour pressure 17.5 mmHg at 20°C MSDS, Sigma-Aldrich, 2009

Vapour density N/A

Water solubility Miscible MSDS, Sigma-Aldrich, 2009

Dissociation constant (pK

a ) 7.49 at 25°C Stumm and Morgan, 1994

Oxidation-reduction potential 0.9 volts Stumm and Morgan, 1994

Corrosivity to the

materials or equipment of normal ship construction Corrosive MSDS, Sigma-Aldrich, 2009

Auto-ignition temperature N/A

Explosive properties If heated, decomposition products may cause containers to rupture or explode MSDS, Siemens water technology, 2000

Oxidizing properties Strong oxidant MSDS, Siemens water technology, 2000

Reactivity Reacts violently with combustible and reducing

materials MSDS, Siemens water technology, 2000

pH in solution 12.31 (6% NaOCl) MSDS, Horizon Chemical Co., Inc, 2004

Colour Clear yellow MSDS, Horizon Chemical Co., Inc, 2004

Odour Pungent Chlorine order MSDS, Horizon Chemical Co., Inc, 2004

Table 3-6: Data related to the aquatic toxicity (HOBr/OBr Species LC 50 (mg/L) Reference

Fish No information was found in major database, but toxicity is expected.

Invertebrate No information was found in major database, but toxicity is expected.

Algae No information was found in major database, but toxicity is expected.MEPC 61/2/12 Annex, page 24

I:\MEPC\61\2-12.doc Table 3-7: Physical and chemical properties of Hypobromous acid

Property Value Reference

Melting point (°C) N.A

Boiling point (°C) N.A.

Flammability (flash point for liquids; °C) Not flammable

Density (20°C; kg/m 3 ) Not located

Vapour pressure (20°C; Pa) 3.06×10 -13 mmHg

Vapour density (air = 1) N/A

Water solubility 1×10 6 mg/L (25 o C)

pH in solution Less than 7

Dissociation constant (pKa) 8.14 Chekanov, et al ., 2007

Oxidation-reduction potential N.A.

Corrosivity to material or equipment N.A.

Reactivity to container material N.A.

Auto-ignition temperature (°C) N.A.

Explosive properties N.A.

Oxidizing properties Oxidizing agent

Surface tension N.A.

Viscosity N.A.

Thermal stability and identity of breakdown products N.A.

Other physical or chemical properties N.A.

Table 3-8: Data related to the acute aquatic toxicity (ozone)

Species Explanation Reference

Channel catfish, Ictalurus punctatus 96 h, LC 50 = 30 μg/L PAN pesticide

database

White perch,

Morone americana 24 h, LC 50 (mean) = 380 μg/L 48 h, LC 50 (mean) = 260 μg/L 96 h, LC 50 (mean) = 200 μg/L PAN pesticide

database Fish

Striped bass (Larvae),

Morone saxatilis 24 h, LC 50 (mean) = 110 μg/L 48 h, LC 50 (mean) = 90 μg/L PAN pesticide

database

Invertebrates Zebra mussel,

Dreissena polymorpha 0.25 h, LT 50 (min) = 360 μg/L
23.9 h, LT 95 (min) = 530 μg/L PAN pesticide

database

Phytoplankton No record foundMEPC 61/2/12 Annex, page 25

I:\MEPC\61\2-12.doc Table 3-9: Physical/chemical properties of ozone

Property Value Reference

Melting point -192.7 ± 2°C HSDB

Boiling point -111.9 0 C HSDB

Flammability Not flammable (NFPA hazard code= 0) MSDS- BOC gases

Density Liquid: 1.614 g/L at -195.4°C HSDB

Vapour pressure > 1atm at 100°F MSDS- BOC gases

Vapour density 2.144 g/l at 0°C MSDS- BOC gases

Water solubility 0.001% at 32°F MSDS- BOC gases

Dissociation constant (pK

a ) Not available

Oxidation-reduction potential > 2700 mV http://www.o3center.org (Web site of O 3 centre)

Corrosivity to the

materials or equipment of normal ship construction Can be corrosive due to oxidation properties. However, fast decomposition does not allow ozone to affect the ship

structure -

Auto-ignition temperature Not available

Explosive properties Pure solid or liquid is highly explosive HSDB

Oxidizing properties Strong oxidant HSDB

Reactivity Reactive with All oxidizable materials HSDB

pH in solution Not available

Colour Colourless gas, dark blue liquid, blue-black crystals HSDB

Odour Characteristic pungent odour in concentrations < 2 ppm HSDB

Table 3-10: Data related to the acute aquatic toxicity (Sodium thiosulfate)

Species LC 50 /EC 50 Reference

Gambusia affinis (fresh water) 24000 mg/L/96hour ECB, IUCLID, 2000

Fish Pimephales promelas (fresh water) 10000 mg/L/96hour Terhaar et al ., 1972

Invertebrates Spirostomum ambiguum (fresh water) 1710 mg/L/24hour Nalecz-Jawecki and Sawicki, 1998

Ankistrodesmus falcatus (fresh water) 64 mg/L/1hour Wodzinski R.S. et al

., 1978 Algae Chlamydomonas

reinhardtii (fresh water) 64 mg/L/1hour Wodzinski R.S. et al

., 1978MEPC 61/2/12 Annex, page 26

I:\MEPC\61\2-12.doc Table 3-11: Data related to the chronic aquatic toxicity (Sodium thiosulfate)

Species LC 50 /EC 50 Reference

Fish Oncorhynchus mykiss 600 mg/L/18 month/no mortality ECB, IUCLID, 2000

Invertebrates No data found in literature

Algae No data found in literature

Table 3-12: Physical and chemical properties of Sodium thiosulfate

Property Value Reference

Melting point (°C) 48 MSDS, Sigma-Aldrich,2009

Boiling point (°C) 100 MSDS, Sigma-Aldrich,2009

Flammability (flash point for liquids; °C) Non-flammable ECB, IUCLID, 2000

Density (20 °C; kg/m 3 ) 1740 kg/m 3 ECB, IUCLID, 2000

Vapour pressure (20 °C; Pa) N/A

Vapour density (air = 1) N/A

Water solubility 293 g/l at 0°C 209 g/l at 20°C http://www.atsdr.cdc.gov/MHMI/mmg184.html

pH in solution 7 in 1% solution MSDS, Sigma-Aldrich, 2009

Dissociation constant (pKa) N/A

Oxidation-reduction potential 0.303 v Morgan and Stumm, 1994

Corrosivity to material or equipment Not

Reactivity to container

material Not

Auto-ignition temperature (°C) Not

Explosive properties Not explosive ECB, IUCLID, 2000

Oxidizing properties No oxidizing properties ECB, IUCLID, 2000

Surface tension N/A

Viscosity N/A

Thermal stability and identity of breakdown products N/A

Other physical or chemical properties N/A

4 TOXICITY ANALYSIS ON DISINFECTION BY-PRODUCTS (G9: 5.2)

Disinfection by-products (DBPs) are produced during the disinfection as a result of reactions between the generated disinfectants and the natural organic matters (NOMs) or inorganic substances in ballast water. Three major groups of DBPs are identified as potential compounds generated during the operation of electro-disinfection in this application:MEPC 61/2/12 Annex, page 27

I:\MEPC\61\2-12.doc .1 Trihalomethanes (THMs);

.2 Haloacetic acids (HAAs); and

.3 Bromate (BrO 3 - ).

The above compounds are regulated by various organizations (e.g., the United States Environmental Protection Agency (USEPA) and the World Health Organization (WHO)) for drinking water. The regulated values are shown in Table 4-1.

The above compounds may pose threats to the receiving environment, especially if the concentrations in discharging ballast water after treatment are considerably high. This section reviews the reported literature to understand the toxicity. The quantification of DBPs in the treated water was conducted and provided in this application (confidential supporting information "SI-list of experiment details").

4.1 Trihalomethane (THM)

Aquatic and mammalian toxicity of THMs reported in literature are summarized in Table 4-2.

According to the table, most of the toxicity effects are related to dose levels that are relatively high (e.g., in the range of mg/L or mg/kg). In addition, at a low level of exposure (e.g., in the range of μg/L), mode of exposure such as drinking becomes important in toxicity. In the electro-disinfection technology in this application, it is found that the level of THMs in discharging ballast water is considerably low. These levels are comparable to the drinking water standards and thus, the expected toxicity upon discharge is very low. Moreover, the effect of dilution at the point of discharge may reduce the concentration available at the receiving environment to even a safer level. The toxicity testing of produced ballast water using the BWMS in this application further confirms that the water toxicity is less concerned.

4.2 Haloacetic acids (HAA)

Aquatic and mammalian toxicity of HAAs reported in the literatures are summarized in

Table 4-3.

Table 4-3 shows that HAAs do have risks to the aquatic life as well as to the mammalian life. However, the concentrations related to the aquatic and mammalian toxicity in these literatures are far above the concentrations found in discharging ballast water treated with the technology in this application, as shown in the risk assessment in this document. In addition, the effect of dilution at the point of discharge reduces the concentration available at the receiving environment to even a safer level. The toxicity testing of produced ballast water presented later further confirms the findings.

4.3 Bromate

Aquatic and mammalian toxicity of bromate reported in literature are summarized in the following table. Table 4-4 shows that bromate may create risks to the aquatic life and to the mammalian life. However the effect concentrations and lethal concentrations on aquatic life are much higher than the concentration of water from the BWMS in our application (see the confidential supporting information "SI-list of experiment details"). Thus, the bromated is not an issue in this application.MEPC 61/2/12 Annex, page 28

I:\MEPC\61\2-12.doc Table 4-1: Regulations for the DBPs in drinking water

Compound USEPA standard for drinking water (Maximum Contaminant Level), mg/L * WHO standard for drinking water, mg/L **

Trihalomethanes (THM) 0.08 (total THMs) Chloroform: 0.3

Bromoform: 0.1

Bromodichloromethane: 0.06

Dibromochloromethane: 0.1

Haloacetic acids (HAA) HAA5 (Monochloroacetic acid,
Dichloroacetic acid, Trichloroacetic acid, Monobromoacetic acid, Dibromoacetic acid): 0.06 Monochloroacetic acid: 0.02

Dichloroacetic acid: 0.05

Trichloroacetic acid: 0.2

Bromate (BrO 3 - ) 0.01 0.01 (provisional)

* Source: Website of US EPA, http://www.epa.gov/safewater/contaminants/index.html#1

, Accessed on 23-02-2010.

** Source: Guidelines for Drinking-water Quality, 3 rd Edition, World Health Organization, Geneva, 2008.MEPC 61/2/12Page 29I:\MEPC\61\2-12.docTable 4-2: Toxicity analysis of THMsDisinfectionby-productTest or nature of study andspeciesEffectReferenceChloroformLeuciscus idus melanotusCarp2 days, LC100 = 176 mg/L US ECOTOXGreen algaePseudokirchneriella subcapitata4 days, EC50= 38 6000 μg/LBrown shrimpPenaeus aztecus4 days, LC50= 26 000 μg/LBromoformAtlantic menhadenBrevoortia tyrannus4 days, LC50= 12 000 μg/LUS ECOTOXChlorodibromomethaneCyprinus carpioLC50; Minimum toxic dose = 31 000 μg/LLC50; Mean toxic dose = 52 000 μg/LPAN PesticideDatabaseTHMRisk assessment on swimmersin swimming poolGeometric mean concentration of THM in chlorinated swimming pool water in thestudy = 32.9±2.4μg/L. Risk level from oral ingestion and dermal exposure < 10-6(negligible risk level defined by USEPA).Risk level from inhalation exposure is7.77×10−4ñ 1.36×10−3. Thus, there is a risk from inhalation exposure route.Additional NotesStandard values used in the assessment:Body weight: male = 69.2 kg, female = 56.4 kg, surface areas of swimmers: male= 1.83 m2, female = 1.58 m2, inhalation rate: 1.0 m3/h, ingestion rate: 25 mL/h,swimming frequency: 120 events/year, swimming duration: 30 years, swimmingexposure time: 1.3 h/ event, average life span: 70 years.Leeet al.,2009THMCancer-related deathsSpecies: Human (populationsize= 5144)Cancer-related deaths in Guastalla, northern Italy between 1987-1999 due todrinking water with increased concentrations of THMs during the period of 1965-1987.Increment in cancers among males: 1.2%Increment in cancers among females: 1.1%Additional NotesThe THM concentration: (chloroform: 33.1μg/L immediately after chlorination,57.1μg/L 24 h after chlorination. Total THM: 39.7μg/L immediately afterchlorination, 70.7μg/L 24 h after chlorination).Lifestyle factors such as smoking were not considered.MEPC 61/2/12Annex, page 30I:\MEPC\61\2-12.docDisinfectionby-productTest or nature of study andspeciesEffectReferenceTCM*BDCM**Human health risk assessmentSpecies: Based on humanepidemiological dataPossibility of bladder and colon cancer in males, as well as developmental effectsand asphyxia amongst infants.Hamidinet al., 2008ChloroformToxicity on miceAt a dose of 300 mg/kg (in olive oil byoral gavage), severe kidney lesions wassound after 24 h.Fanget al.,2008BDCMNeurotoxicological evaluationSpecies used: RatsDose: 9, 27, and 72 mg/(kg day), through drinking water for 6 months.Outcome: Neurotoxicity may not be a concernMoseret al.,2007CDBM***Toxicity/CarcinogenicitySpecies: Rats/MiceAdministered through gavage (in corn oil) for 13 weeks and 2 years.13 weeks study:Dose: 0, 15, 30, 60, 125, and 250 mg/kgObservations:Rats: At 250 mg/kg, hepatic and renal toxicity production and increment inmortality among both male and female (9/10 were dead)Mice: At 250 mg/kg, hepatic and renal toxicity production (in male)2 years (chronic) study:Dose: Rats ñ 0,40, 80 mg/kg, Mice ñ 0, 100 mg/kgObservations:Rats:Male ñ Survival does not affected by treatment. Body weight reduced at 80 mg/kgdose level. Fatty change and cytoplasmic changes occurred in the liver at bothdose levels.Female ñ Survival does not affected bytreatment. Fatty change and cytoplasmicchanges occurred in the liver at both dose levels.Mice:Male ñ Low survival relative to control, Low body weights, Non-neoplastic hepaticlesions (nephrosis and hepatocytomegaly), nephrosis.Female ñ Low body weights Non-neoplastic hepatic lesions (calcification and fattychange)Dunnicket al., 1985MEPC 61/2/12Page 31I:\MEPC\61\2-12.docDisinfectionby-productTest or nature of study andspeciesEffectReferenceBDCMToxicity (acute)Species: Female Rats/miceAdministered for 5 days by gavage.Rats: 75, 150, 300 mg/kg bw/dayAt 300 mg/kg bw/day, 2/6 rats were dead at day 5. Renal and hepatic damagesoccurred at 300 mg/kg bw/day. BDCM is Hepatotoxic and nephrotoxic.Mice: 75, 150 mg/kg bw/day.BDCM is weakly Hepatotoxic and nephrotoxic.ThorntonManninget al., 1994BDCM andchloroformAcute renal and hepatic Effects(comparison betweenchemicals)Species used: RatsOral gavage (single dose), 0.0, 0.75, 1.0, 1.5, 2.0, 3.0 mmol/kg (BDCM or CHCl3).Both compounds were hepatotoxic at 24 after the administration of≥1.5 mmol/kg.BDCM was significantly elevate the serum hepatotoxicity than CHCl3after 48 h.BDCM was more toxic to liver and kidney compared to CHCl3.Lillyet al.,1997Notes:*TCM: Trichloromethane;**BDCM: Bromodichloromethane;***CDBM: Chlorodibromomethane.MEPC 61/2/12Annex, page 32I:\MEPC\61\2-12.docTable 4-3: Toxicity analysis of HAAsDisinfectionby-productTest or nature ofstudySpecies Effect ReferenceLemna gibba7 d, EC50= 8.6 mg/L, Endpoint= plant growth rateMyriophyllumspicatum4 d, EC50=13.3 mg/L, Endpoint= plant lengthMCAA1Toxicity onaquatic lifeMyriophyllumsibiricum4 d, EC50= 24.3 mg/L, Endpoint= plant lengthHansonet al.,2002aMCAA: 7 d, EC50= 20.7 mg/L, Endpoint= Growth rateDCAA: 7 d, EC50= 555.2 mg/L, Endpoint= Growth rateLemna gibbaTCAA: 7 d, EC50= 864.3 mg/L, Endpoint= Growth rateMCAA: 14 d, EC50= 10.9 mg/L, Endpoint= Plant lengthDCAA: 14 d, EC50= 112.5 mg/L, Endpoint= Plant lengthMyriophyllumspicatumTCAA: 14 d, EC50= 140.3 mg/L, Endpoint= Plant lengthMCAA: 14 d, EC50= 8.0 mg/L, Endpoint= Plant lengthMCAADCAATCAA3Toxicity onaquatic lifeMyriophyllumsibiricumTCAA: 14 d, EC50= 119.9 mg/L, Endpoint= Plant lengthHansonet al.,2004TCAAToxicity onaquatic lifeMyriophyllum spp.0.05, 0.5, 3, and 10 mg/l of TCAA for 35 days.No biologically significant effects were found and thus TCAA does not pose aconsiderable risk on the species up to 10 mg/LHansonet al.,2002bPseudokirchneriellasubcapitata72 h EC50(Growth) = 16.2 mg/L; 72 h EC50(Biomass) = 4.9 mg/L72 h NOEC(Growth) = 3.00 mg/L; 72 h NOEC (Biomass) = 3.00mg/LTCAAToxicity toaquatic lifeScenedesmussubspicatus72 h EC50(Growth) = 67.9 mg/L; 72 h EC50(Biomass) = 4.7 mg/L72 h NOEC (Growth) = 3.00 mg/L;72 h NOEC (Biomass) = 3.00mg/LRobertset al.,2010MBAA4DBAA5Acute toxicity/ RatsAcute toxicity (gavaged, most deaths within 48 h)MBAA: LC50=177 mg/kg; DBAA: LC50=1737 mg/kgLinderet al.,1994MEPC 61/2/12Page 33I:\MEPC\61\2-12.docDisinfectionby-productTest or nature ofstudySpecies Effect Referenceacute spermatotoxicityAcute spermatotoxicityMBAASingle dose, 100 mg/kg: No reproductive effects14 daily doses, 25 mg/kg: No reproductive effectsDBAASingle dose, 1250 mg/kg:•17% drop of serum testosterone after 2 d (recovered after 14 d)•Affected the sperm motion (post treatment day 14 and day 28 )•Degenerative flagellar changes in cauda sperm (day 14)•Abnormal sperm head shapes (day 28)DBAADCAABCAA6Mortality,dysmorphology/growthretardationRat Embryos48 h exposure during culturingDBAA: 400μM, 39% mortality, decreases in somite number and head lengthDCAA: 5000μM, 23% mortality, malformations, and decreases in somitenumber, crown-rump length, head length and developmental scoreBCAA: 300μM, 13% mortality, decreased DEVSC, somitenumber and head lengthAndrewset al., 2004DCAATCAAHuman healthrisk assessmentBased on humanepidemiological dataPossibility of bladder and colon cancer in males, as well as developmentaleffects and asphyxia amongst infants.Hamidinet al., 2008BCAAReproductivetoxicity in miceMice (juvenile andadult)Juvenile males: 14 d, gavage0-72 mg/kg-No significant variation in average body weight, No significantdifferences in the relative testis weights216 mg/kg-Significantly small body weight, Physically and behaviorallynormal, No significant differences in the relative testis weights. 14 weeksafter the dosing period and 40 d mating period- No significant differences inrelative weights reproductive organs, no significant differences in reproductiveparameters648 mg/kg-Lethally toxicTullyet al.,2005MEPC 61/2/12Annex, page 34I:\MEPC\61\2-12.docDisinfectionby-productTest or nature ofstudySpecies Effect ReferenceAdult males: 14 d, gavage0-216 mg/kg-No signs of general toxicity or significant changes in bodyweight, No significant effect on relative testis, epididymal, or seminal vesicleweight. 40 d mating period after the 14 d dosing- No effect on the libido of theadult males, For 72 and 216 mg/kg, significant reduction in the numbers oflitters/plug, litters/male and fetuses/male for the first 10 d of mating period.40 well-characterized genes differentially expressed in a dose responsivemanner.DCAADBAABCAADysmorphogenesis in mice- shortterm exposureMice (Gestation day8, CD-1 mouseconceptuses)Reported increased incidences of dysmorphic (Cranial neural tubeclosure defect, Prosencephalichypoplasia, Pharyngeal archhypoplasia, Eye, Heart) embryos:DCAA: 11,000 μM, 6 or 26 h exposureDBAA: 300 μM, 26 h exposureBCAA: 300 μM, 3, 6 or 26 h exposureHunteret al.,2006DCAATCAASubchronictoxicity (90 d)Rats0-5000 ppm in drinking water, 90 d.DCAA:500 and 5000 ppm- terminal body weights were significantly reduced.Total serum protein levels were significantly depressed in all animals.TCAA:Decreased body weights were apparent, but not statistically significant.No consistent differences were seen in serum chemistry parameters.Matheret al.,1990MEPC 61/2/12Page 35I:\MEPC\61\2-12.docDisinfectionby-productTest or nature ofstudySpecies Effect ReferenceDCAA Carcinogenicity RatsDoses administered through drinking water.Exposure period: 103 weeksMean daily concentration and dose: 0.05 g/L (3.6 mg/kg bw/day),0.5 g/L (40.2 mg/kg bw/day), 1.6 g/L (139 mg/kg bw/day)Observations:Toxic to nervous system, liver andmyocardium while treatment-relatedneoplastic lesions were observed only in the liver.Considerable increment in carcinogenicity (hepatocellular carcinoma) wasobserved at the concentration of 1.6g/l.At 100 weeks, the concentration of 0.5g/l increased hepatocellular neoplasia,(carcinoma and adenoma).No observed effects level = 0.05g/l (3.6mg/kg/day).DeAngeloet al., 1996DBAAToxicity/CarcinogenicityRats/Mice2 weeks,Concentrations in drinking water: 0, 125, 250, 500, 1000, and 2000 mg/LRats:Daily dose 0-270 mg/kg. No adverse effects on water consumption,survival, body weight gain, clinical signs. 12-44% increment in lever weight.Male- If concentration > 250 mg/L, incidences of diffuse cytoplasmic alterationof hepatocytes were increased. For concentration > 500 mg/L, relative testisweights reduced by 8-15%.Female- If concentration was 2000 mg/L, incidences of diffuse cytoplasmicalteration of hepatocytes were increased.Mice:Daily dose 0- 370 mg/kg. No adverse effects on water consumption,survival, body weight gain, clinical signs.Male- Concentration of 1000 ñ 2000 mg/L increased the incidences of thymicatrophyFemale- Concentration of 2000 mg/L increased the incidences of thymicatrophyMelnicket al.,2007MEPC 61/2/12Annex, page 36I:\MEPC\61\2-12.docDisinfectionby-productTest or nature ofstudySpecies Effect Reference3 monthsConcentrations in drinking water - 0, 125, 250, 500, 1000, and 2000 mg/LRats:Daily dose 0-181 mg/kg. At concentration of 2000 mg/L, 10% reductionin body weight. No effects on water consumption and survival. Up to 40%increment in lever weight. Increases in centrilobular to midzonal glycogen-likehepatocellular cytoplasmic vacuolization.Male- 50% reduction in testis weight and reduction in sperm motility andsperm concentrations at exposure to 2000 mg/L.Mice:Daly dose level- 0-260 mg/kg. At concentration of 2000 mg/L, 10%reduction in body weight. No effects on water consumption and survival.2 yearsConcentrations in drinking water - 0, 50, 500, and 1000 mg/LRats:No effect on survival. 5-15 % reduction in mean body weight if exposedto 500-1000 mg/L. Neoplastic effects (Increased incidences of mononuclearcell leukaemia).Male- Neoplastic effects (Induction of malignant mesotheliomas).Non-neoplastic lesions (minimal to mildcystic degeneration in the liver).Female- Non-neoplastic lesions (alveolar epithelial hyperplasia).Mice:No effect on survival. Lever neoplastic effects (increasedhepatocellular adenoma or carcinoma incidences).Male- Lever neoplastic effects (increased hepatoblastoma incidences).In 500 mg/L group, increment in Incidence of alveolar/ bronchiolar adenomaor carcinoma.1MCAA: Monochloroacetic acid;2DCAA: Dichloroacetic acid;3TCAA: Trichloroacetic acid;4MBAA: Monobromoacetic acid;5DBAA: Dibromoacetic acid;6BCAA: Bromochloroaceticacid.MEPC 61/2/12Page 37I:\MEPC\61\2-12.docTable 4-4: Toxicity analysis of BromateDisinfectionby-productTest or nature ofstudySpecies Effect ReferenceGlenodinium halli(Dinoflagellate), marinephytoplanktonChronic studies (exposure period is not specified), EC50>13.6mg BrO3-/LDaphnia magna, (fresh water,invertebrate)Acute, 48-h LC50of 179 mg BrO3-/LPolycelis nigra,(fresh water,invertebrate)48-h LC50of 2258 mg BrO3-/LMysid shrimp(altwater, invertebrate)Acute, 24-h LC50of 176 mg BrO3-/LLeiostomus xanthurus(juvenile), (Saltwater fish)2 days, LC50of 518 mg BrO3-/L7 days, LC50379 mg BrO3-/LBrO3-Review on effectof BrO3-on aquaticorganismsMorone saxatilis (pro-larvae),(Saltwater fish)2 days, LC50of 605 mg BrO3-/L7 days, LC50of 197 mg BrO3-/LHutchinsonet al,1997BrO3-Acute toxicityCrassostrea virginica, larvae(Oyster)1 mg/L initial dose, 48 h,> 90% mortalityStewartet al.,1979BrO3-Alterations inprotein and geneexpression in ratkidneyRatsExposed to potassium bromate (drinking water) atconcentrations of 0, 1, 20 and 400 ppm.Alteration in mitochondrial function, glycolysis andmitochondrial gene transcripts.Ahlbornet al.,2009MEPC 61/2/12 Annex, page 38

I:\MEPC\61\2-12.doc 5 TOXICITY TESTING OF THE TREATED BALLAST WATER (G9: 5.2)

The objective of the development of the BWMS is to protect our marine environment. In order to effectively disinfect the unwanted/harmful organisms/micro-organisms, the Active Substances are generated by the electrochemical reactor a nd used. It has been demonstrated that the BWMS can achieve the goal; one of the evidences is shown in the confidential supporting information "SI-Disinfection study" on the disinfection of E. coli and Intestinal Enterococci

. Although the working mechanism of electrolysis has being widely reported and accepted, the studies of treated effluent in terms of eco-toxicity and environmental impact are still very important, because the performance of electrolys is systems different from one to the other, which produces different types and amounts of toxicants. As such, a series of studies has been conducted by various research institutions shown in the confidential supporting information "SI-list of organizations in R&D of BWMS". The studies were: 1. measurements of concentrations of various environmental relevant compounds (e.g., THMs and HAAs); 2. eco-toxicity testing by fish, invertebrate and alga. The results are summarized as follows.

5.1 pH

pH is one of the most important physicochemical parameters of the seawater. A suitable pH range of the discharged ballast water is essential to ensure the safety of ecosystem of receiving water and the safety of the ships.

An experiment was conducted with various energy inputs applied to an electrochemical reactor, in which chlorine with different concentrations was generated. The pH was measured by a pH meter (ORION 920Aplus). Prior to the measurements, the pH meter was calibrated using pH standard solutions (pH = 4, 7, and 10). As shown in Figure 5-1, the pH in the effluent is quite similar to that in the influent.

An experimental study on change in pH was conducted in February 2010; it was observed by a surveyor of the ABS. The result showed that, when the pH of the influent seawater is 7.97, the pH of the effluent seawater was 7.68 ("SI-list of experiment details").

According to the chemical reactions shown early , the pH of the seawater after the reactor may be lower than that of the influent. However, due to the short retention time of the seawater in the reactor, the decrease in pH of the seawater is less significant. In addition, there are some anionic substances in the solution (e.g., carbonate). They have buffering capacities, which can neutralize hydrogen ions. As a result, the decrease in the pH is less significant and no further increase in corrosion is expected.

01234 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0pH

Total chlorine concentration, ppm Influent pH Effluent pH

Figure 5-1: Change in solution pH as a function of chlorine concentrationMEPC 61/2/12 Annex, page 39

I:\MEPC\61\2-12.doc 5.2 Total residual oxidant

As the above analysis, the TRO (measured as Cl 2 ) is mainly contributed by active chlorine, which plays an important role for the disinfection of the ballast water. A high concentration of active chlorine may be toxic to the aquatic organisms in the receiving water. Hence, the active chlorine in the discharged ballast water must be controlled under a very low level (0.1 mg/L).

The initial active chlorine concentration of the seawater in ballast tanks is 12 mg/L, so that organisms and micro-organisms can be killed. After the disinfection, most of chlorine is consumed to a low value (e.g., 2-3 mg/L). Under such a level, the decay of active chlorine occurs in the ballast tank quickly (referred to the confidential supporting information "SI-Chlorine decay in the ballast water"). In addition, the active chlorine will be neutralized by sodium thiosulfate during deballasting. The amount of the dosed sodium thiosulfate is accurately controlled according to the level of total residual oxidants in ballast water, which is monitored by an online analyzer.

According to the dosage given in the confidential supporting information "SI-Description for the BWMS", the maximum allowable concentration of active chlorine in the discharged ballast water after neutralization is 0.1 mg/L. Under such a low level of TRO, the safety of the aquatic organisms in the receiving water can be ensured.

An experiment was conducted under the ABS observation and verified by staff from T‹V S‹D PSB Pte Ltd. The TRO (free chlori ne concentration) was measured by T‹V S‹D
PSB Pte Ltd; the value was 0.01 mg/L, far less than 0.1 mg/L. The risk of treated water in the marine environment is expected to be insignificant.

5.3 Disinfection by-products

Disinfection by-products may be generated during the ballast water treatment process using the BWMS in this application. Three majo r groups of DBPs, including trihalomethanes (THMs), haloacetic acids (HAA) and bromate (BrO 3 - ), are identified as potential compounds generated during the proposed electro-disinfection process.

An experiment was conducted with two types of contaminated seawater with TOC of 4.87 and 4.88 mg/L and TSS of 134 and 120 for the water with 24 and 35 PSU, respectively. The water was not filtrated, in order to simulate the worse scenario. The water with the Active Substance (chlorine) after a period of time (e.g., after 2 hours termed as Day 0, 24 hours termed as Day 1, and 5 days termed as Day 5) was collected for the analysis of various parameters stated below (e.g., THMs, and HAAs).

5.3.1 Trihalomethanes (THMs)

Trihalomethanes (THMs) are chemical compounds in which three of the four hydrogen atoms of methane (CH 4 ) are replaced by halogen atoms. As discussed previously, THMs are environmental pollutants and some of them are considered as carcinogenic. Chloroform/ trichloromethane (CCl 3 H), bromodichloromethane (CBrCl 2 H), chlorodibromomethane (CClBr 2 H) and bromoform/tribromomethane (CBr 3 H) are four THMs, which may exist and are concerned in the discharged ballast water treated by the BWMS. The results of THMs are

showed in Table 5-1.MEPC 61/2/12 Annex, page 40

I:\MEPC\61\2-12.doc Table 5-1: Concentration of THMs in treated water

24 PSU 35 PSU

Parameter Day 0 (ppb) Day 1 (ppb) Day 5 (ppb) Day 0 (ppb) Day 1 (ppb) Day 5 (ppb)

Trichloromethane N.D N.D N.D N.D N.D N.D

Bromodichloromethane N.D N.D N.D N.D N.D N.D

Dibromochloromethane 13.7 10.1 N.D 14.1 7.67 N.D

Tribromomethane 128.7 114.3 10.9 153.1 112.3 23.8

* N.D means under detection limit.

+ Measured by PUB (Singapore) according to ISO/IEC 17025; Report #CAWT 100000221

.

The THMs may have adverse health effects at high concentrations, and many governments set limits on the amount permissible in drinking water. In the United States, the EPA limits the total concentration of chloroform, bromoform, bromodichloromethane, and dibromochloromethane to 80 ppb in drinking water. The results, summarized in Table 5-2, show the total trihalomethanes in the treated ballast water after 5 days are less than 80 μ g/L, which is lower than what is required for the drinking water.

5.3.2 Haloacetic acids

Haloacetic acids (HAAs) are carboxylic acids in which a hydrogen atom in acetic acid is replaced by a halogen. HAAs are a common undesirable disinfection by-product in water disinfected by chlorine. The concentrations of HAAs in the treated ballast water are shown in

Table 5-2.

Table 5-2: Concentration of HAAs in treated water

24 PSU 35 PSU

Parameter Day 0 (ppb) Day 1 (ppb) Day 5 (ppb) Day 0 (ppb) Day 1 (ppb) Day 5 (ppb)

Monochloraacetic acid N.D N.D N.D N.D N.D N.D

Dichloroacetic acid N.D N.D N.D N.D N.D N.D

Trichloroacetic acid 6.48 6.66 11.2 6.68 N.D 18.7

Monobromoacetic acid 12.5 5.09 N.D 8.31 N.D 11.4

Dibromoacetic acid 73.0 64.4 106.1 101.4 76.3 175.8

Tribromoacetic acid 190.8 89.9 146.4 308.8 160.6 373.1

Bromochloroacetic acid 5.55 5.14 6.98 7.53 N.D 12.1

Dibromochloroacetic acid 32.9 15.7 23.0 50.8 17.9 52.5

Dichlorobromoacetic acid N.D N.D 5.83 5.18 N.D 10.6

* N.D means under detection limit.

+ Measured by PUB; Report #CAWT 100000221.

5.3.3 Bromate (BrO 3 - )

The bromate concentration is under detection limit (0.005 mg/L) as shown in Table 5-3. As bromate is formed in the presence of ozone, it can be concluded that the ozone production by our BWMS is negligible.MEPC 61/2/12 Annex, page 41

I:\MEPC\61\2-12.doc Table 5-3: Concentration of bromate (BrO 3 - )

Water Day 0 (mg/L) Day 1 (mg/L) Day 5 (mg/L)

24 PSU N.D * N.D N.D

35 PSU N.D N.D N.D

* N.D means under detection limit. + Measured by PUB; Report #CAWT 100000221.

5.3.4 Analytical methods at environmentally relevant concentrations (G9: 4.2.1.5)

The important analytical methods at environmenta lly relevant concentrations are shown in Table 5-4. The full list of analytical methods and detection limits can be found in the confidential supporting information "SI-list of experiment details".

Table 5- 4: Analytical methods at environmentally relevant concentrations

Chemicals Methods

Trihalomethanes (THMS) EPA 8260B

Haloacetic acids (HAA) EPA 552

Bromate ion EPA 300.1

5.4 Eco-toxicity testing

To evaluate the potential aquatic toxicity level of the treated ballast water by using the proposed electro-disinfection technology, the aquatic toxicity tests of marine fish, marine invertebrate and marine algae for ballast water treated by the BWMS have been carried out. The procedures follow to the requirements of the "Procedure for approval of ballast water management systems that make use of Active Substances (G9)", and are based on the Organisation for Economic Co-operation and Development (OECD) Guidelines.

5.4.1 Aquatic toxicity test on marine fish

.1 Japanese Medaka ( Oryzias latipes )

It has been demonstrated that Japanese Medaka is very sensitive to toxins (Belanger, et al ., 1990; Davis, et al ., 2002; Sun et al ., 2009). Hence, it was selected in the ecotoxicity study. Acute toxicity testing (96 hours) and chronic toxicity testing (30 days) of the treated ballast water by using the Japanese Medaka ( Oryzias latipes ) were conducted in a laboratory of Yantai Institute of Coastal Zone Research of Chinese Academy of Sciences, China. The institute has a group of experts in environmental assessment (marine). Each testing used 0, 6.25%, 12.5%, 25%, 50% and

100% of the ballast water. It was found that the ballast water did not have any toxic effect on the marine fish in both acute and chronic toxicity studies.

.2 Abudefduf saxatilis

Acute toxicity testing (96 hours) and chronic toxicity testing (10 days) of the treated ballast water by using the Abudefduf saxatilis were conducted in a laboratory of T‹V S‹D PSB Pte Ltd. The laboratory is certified withMEPC 61/2/12 Annex, page 42

I:\MEPC\61\2-12.doc ISO/IEC 17025. The fish was selected as it normally lives in the coastal area (~20 metres depth). Each testing used 0, 6.25%, 12.5%, 25%, 50% and

100% of the ballast water. It was found that the ballast water did not have any toxic effect on the marine fish in both acute and chronic toxicity studies.

5.4.2 Aquatic toxicity test on marine invertebrate

.1 Penaeus monodon

The Penaeus monodon was selected as it is very sensitive to toxins (Benzie, 1997; Chen and Lin, 2001; Tsai and Chen, 2002). Acute toxicity testing (96 hours) and chronic toxicity testing (30 days) of the treated ballast water by using the Penaeus monodon were conducted in a laboratory of Yantai Institute of Coastal Zone Research of Chinese Academy of Sciences, China. Each testing used 0, 6.25%, 12.5%, 25%, 50% and 100% of the ballast water. It was found that the ballast water did not have any toxic effect on the shrimp in both acute and chronic toxicity studies.

.2 Stenopus hispidus

Acute toxicity testing (96 hours) and chronic toxicity testing (10 days) of the treated ballast water by using the Stenopus hispidus were conducted in a laboratory of T‹V S‹D PSB Pte Ltd. The shrimp was selected as it is sensitive to toxins. Each testing used 0, 6.25%, 12.5%, 25%, 50% and 100% of the ballast water. It was found that the ballast water did not have any toxic effect on the marine shrimp in both acute and chronic toxicity studies.

5.4.3 Algae growth inhibition testing

Dunaliella primolecta (UTEX LB 1000), a marine micro-alga, has been used in toxicity study for seawater (Bentley-Mowat and Reid, 1977; Videau, et al ., 1979; Pun, et al ., 1995). It was used in the inhibition testing for the treated water from the BWMS in this application. The water used was 0, 6.25%, 12.5%, 25%, 50% and 100% ballast water treated by the BWMS. It was found the there was no difference between the controlled water (i.e. clean seawater) and the treated ballast water. The experimental details can be found in the confidential testing report "Algae growth inhibition study". It is worthy to note that the NaOCl is rated as a highly toxic chemical to Dunaliella primolecta (LC 50 = 400μg/L) in PAN pesticide database.

5.4.4 Ecox toxicity testing

The Eclox toxicity testing technology is an inte rnationally recognized analysis technique on toxicity of water (Scully, 1998; Ellis, 2000; Benfenati, et al ., 2003). It was used to evaluate the level of toxicity of the treated ballast water. The obtained results illustrate there is no difference of the level of toxicity of the treated ballast water by the proposed BWMS and untreated ballast water. The study was conducted by the NUS staff in the PUB Lab, Singapore. Please refer to the confidential laboratory report "Eclox toxicity testing report".

6 RISK ASSESSMENT

6.1 Safety of ship (G9: 6.3)

Fire/explosion can cause immediate damage a ship partially or fully. Hydrogen gas is the only gas that may cause fire/explosion. On the other hand, corrosion in ballast water tank may slowly consume the steel/metal of the tank, which may damage the tank and eventually the ship. As such, both hydrogen gas and corrosion due to the operation of the BWMS were

studied with the results summarized as follows.MEPC 61/2/12 Annex, page 43

I:\MEPC\61\2-12.doc 6.1.1 Gaseous species

.1 Hydrogen gas

There are three factors that can lead to the fire/explosion; they are fuel(s), oxidizer(s) and ignition source(s). Oxygen as an oxidizer is well available; 21% of air is oxygen. The ignition sources in a ship are available. Wiring of equipments (e.g., motor), smoking, lightning, and spark can act as ignition sources and they are all available when a ship is operated. The minimum ignition energy is 0.018 mJ for hydrogen gas (Crowl and Louvar, 2002).

In the BWMS in this application, hydrogen gas is the only one that may cause fire/explosion. The lower flammability limit (vol. % fuel in air) of hydrogen is 4% and the upper flammability limit is 75% according to appendix B of "Chemical Process Safety, Fundamentals with Applications" (P. 568) (Crowl and Louvar, 2002).

An experiment of electrolysis by our BWMS was conducted. The concentration of hydrogen gas from the BWMS was experimentally determined by T‹V S‹D PSB PTE LTD . The detection limit is 0.5% for the hydrogen gas. It was found the concentration of H 2 was under the detection limit (less than 0.5%). The system is fully sealed and no gas leakage is expected. Thus, the possibilities of explosion can be neglected. The confidential supporting information "SI-Calculation of hydrogen concentration" and "SI-list of experiment details" provides more detailed analysis of the risk of hydrogen gas.

.2 Chlorine gas

Chlorine is produced through the electrolysis reaction shown early. It can rapidly dissolve in the water forming HClO and ClO
- . However, there could be chlorine gas left in the ballast water tank, which could create hazard problems even its concentration is low. An experiment of electrolysis by our BWMS was conducted. The concentration of chlorine gas from the BWMS was determined by T‹V S‹D PSB PTE LTD . The detection limit is 0.1% for the gas. It was found the concentration was under the detection limit (less than 0.1%) (the confidential supporting information "SI-list of experiment details"). As the whole electrochemical reactor is fully sealed, low risk in the inhalation of toxic gas to the crew is expected. Furthermore, an optional ventilation system is used for the ballast water tank.

6.1.2 Corrosion

The corrosion study for the ballast water treated with active chlorine was carried out to evaluate the effects of the BWMS on the ship safety. The testing includes uncoated steels and anti-corrosion coated steel supplied by the Neptune Orient Lines (Singapore). See below:

Uncoated steel specimen

The corrosion rates of uncoated steels were conducted using the method of mass loss (ASTM G1-90). Figure 6-1 shows that the corrosion morphology of the uncoated steel specimen subjected to the chlorinated water is the same to those subjected to nature seawater (no chlorine).MEPC 61/2/12 Annex, page 44

I:\MEPC\61\2-12.doc Figure 6-1: Comparison of corrosion morphology of uncoated steel specimens: experiments conducted for 1.5 months, initial chlorine concentration = 10 mg/L,

chlorine concentration after 1 month = 0.

The corrosion rates measured for the two specimens (uncoated steel in seawater and chlorinated seawater) are listed in Table 6-1. As shown, the corrosion rate of the uncoated steel specimens after one month in the chlorinated seawater with an initial TRO of 9.2 mg/L is 0.11 mm/year, slightly higher than that in the natural seawater (not chlorinated). The corrosion rates of both after 4 months are very similar to each other. The results from one and four-month corrosion studies indicate that the seawater treated by the electrolysis device of the BWMS in this application have an insignificant impact on the corrosion that are similar to those reported by Song et al . (2005) and Dang (2005).

Table 6-1: Corrosion rate results for uncoated steel specimens

Duration time 1 month 4 month

Natural Seawater 0.09 0.07 Corrosion Rate (mm/yr) Treated Seawater 0.11 0.08

* Initial chlorine concentration was 10 mg/L. The chlorine concentrations after 1 month

and 4 months were 0. 

### Before corrosion After corrosion Chlorinated Seawater

Natural SeawaterMEPC 61/2/12 Annex, page 45

I:\MEPC\61\2-12.doc Anti-corrosion coated steel specimens

It has been well documented that seawater used as ballast can cause corrosion. Thus, the ballast water tank must be painted by anti-corrosion paint before the ballast water is stored for balancing purpose. The corrosion is starte d from damaging the paint chemically before that of metal happens. When the BWMS in this application is used, active chlorine could enhance the corrosion. Two studies (morphology and FTIR) were conducted with the results

shown as follows:

Table 6-2: Corrosion morphology of coated steels

Duration time Before experiment 1 month

Not chlorinated

seawater

Coated steels

Chlorinated seawater

* Initial chlorine concentration was 10 mg/L. Chlorine concentration after 1 month was 0.

An experiment on the anti-corrosion coated steel specimens has been tested for one month. The corrosion morphology shown in Table 6-2 indicates no corrosion on the specimens with the protection of anti-corrosion coating in both chlorinated and not-chlorinated seawater.

The anti-corrosion paint is essentially organic (polymer). Corrosion will occur first on the paint then the steel. Any oxidation (i.e. corrosion) on the paint will change its chemistry. The Fourier transform infrared spectroscopy (FTIR) , a highly sophisticated scientific analytical instrument was used to determine the changes in chemistry of the paint (organic/polymer compound). The peaks in the FTIR curve result from the vibration frequency of one or few typical functional groups on an organic substance (paint in this case).

As shown in Figure 6-2, the vibration frequency values of all these specimens at any wavelength are exactly the same. This indicates that the usage of Active Substance generated from our BWMs in this application does not further strengthen/increase the corrosion. The experimental details are given in the confidential supporting information "SI-FTIR for corrosion study".MEPC 61/2/12 Annex, page 46

I:\MEPC\61\2-12.doc 0 1000 2000 3000 4000 5000 Control sample Chlorinated seawater sampleTransmittance (%)



### Wavelength (nm) Raw seawater sample

Figure 6-2: Corrosion study by FTIR

The reasons causing insignificant differences in the corrosion of uncoated and coated steel specimens in the non-chlorinated seawater and chlorinated seawater is mainly due to the decay in the active residual chlorine. As shown in the confidential supporting information "SI-Chlorine decay in the ballast water", the decay of chlorine is relatively faster in the contaminated water. For example, the residual chlorine concentration becomes zero after 4 days when the initial concentration is 3.6 mg/L. Since the chlorine disappears after 1 to 2 weeks, the water originally chlorinated is the same as the non-chlorinated water. Hence, the corrosion in the treated ballast water is very similar to that in the seawater.

The testing for both uncoated and coated specimens is being conducted according to Procedure (G9) of 6-month duration. The final testing results will be provided in the application dossier for the Final Approval.

6.2 Human and environment (G9: 6.4)

The concentrations of Active Substances and DBPs from the treated water were measured by the CAWT (PUB). They were used to evaluate the effects of treated ballast water on the human and the marine environment. The assessment was conducted by the DHI, an independent research company, which has no relationship in the development of the BWMS in this application. The summary of risk assessment by the DHI is given below as well as in the confidential supporting information "SI-DHI letter". The detailed information is available in the confidential information "SI-DHI risk assessment report".MEPC 61/2/12 Annex, page 47

I:\MEPC\61\2-12.doc 6.2.1 Overall conclusion on the environmental risk assessment

The risk assessment is performed as a conservative estimation using the estimated maximum concentrations of potentially released Active Substances and by-products inside the harbour.

Based on these conservative estimations , the following conclusions are drawn:

.1 The concentration of whole effluent in the OECD commercial harbour scenario amounts to 6.55 ml/L. Acute ecotoxicity tests performed on algae, crustaceans and fish indicate no toxic effects up to 1000 ml/L (100% undiluted treated ballast water).

.2 Based on a PEC/PNEC evaluation, trichloroacetic acid (RQ = 1.1), monobromoacetic acid (RQ = 3.4) and hypochlorite and hypobromite (RQ = 1.9) poses a potential risk to the environment, which is based on the most conservative assumptions

.

.3 The assessment factor used for derivation of the PNEC value for

trichloroacetic acid and monobromoacetic acid is 10,000 as only acute toxicity data could be found. This means that a very conservative approach has been provided. For the Final Approval of the Blueseas Ballast Water Management System more thorough toxicity data (chronic toxicity data) should if possible be applied for the derivation of the PNEC values.

.4 The calculations on hypochlorite and hypobromite are very conservative and based on the detection limit of 0.01 mg/L.

.5 None of the other evaluated substances show a PEC/PNEC ratio above 1.

.6 Based on the evaluated fate of the substances, none of the substances are expected to accumulate in sediments.

6.2.2 Overall conclusion on the human exposure assessment

Exposure to humans from the TRC and its degradation products may occur due to:

.1 Inhalation from exhaust air emitted from the ballast tanks (occupational). The predicted concentration in the air for inhalation is far below existing OEL-values. Thus, no risk to the crew on the ships is expected.

.2 Exposure via the environment due to eating the fish from the harbour area and due to swimming in the harbour area. As the predicted daily uptake considering all uptake routes simultaneously is far below existing TDI values, no risk to the public from the TRC and its degradation products is expected.

7 ASSESSMENT REPORT (G9: 4.3, 4.2.3)

7.1 Experimental data

The experimental data provided in the application were produced based on standard protocols which are internationally applicable. All experiments were conducted at the presence of a third-party surveyor from the ABS

.MEPC 61/2/12 Annex, page 48

I:\MEPC\61\2-12.doc The concentrations of various parameters were measured by the internationally recognized laboratories with relevant certifications (ISO/IEC 17025). The experiments and testing were performed according to the rigorous quality control/quality assurance programme. The reports of testing (e.g., by T‹V S‹D PSB Pte Ltd. and Centre for Advanced Water Technology of
Public Utilities Board, Singapore) and environmental/risk assessment by DHI are provided with the application dossier to the committee for assessment (confidential supporting information).

7.2 Literatures for toxicity analysis

All the literatures cited in this application are from peer-reviewed journals, books, government reports, and databases of international organizations (e.g., ECB, EPA, and WHO). The sources of data are trustworthy and thus the collected information based on literatures for assessment and discussion in this application is reliable.

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