I: \ MEPC \ 56 \ 2

- 3.doc For reasons of economy, this document is printed in a limited number. Delegates are

kindly asked to bring their copies to meetings and not to request additional copies. INTERNATIONAL MARITIME ORGANIZATION

IMO 

# E

MARINE ENVIRONMENT PROTECTION

COMMITTEE
56th session Agenda item 2 MEPC 56/2/3 6 April 2007 Original: ENGLISH

HARMFUL AQUATIC ORGANISMS IN BALLAST WATER

Ba sic Approval of Active Substances used by Resource Ballast Technologies System (Cavitation combined with Ozone an d Sodium Hypochlorite treatment )

Submitted by South Africa

SUMMARY

Executive summary: This document contains non

- confidential informatio n for Basic Approval of Active Substance s used by Resource Ballast Technologies S ystem (Cavitation combined with Ozone and Sodium Hypochlorite tre

atment) in accordance with the P rocedure for approval of Ballast Water Management Systems that make use of

Act ive Substance s (G9) adopted by resolution MEPC.126(53). 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.

Action to be taken: P aragraph 6

Related documents: BWM /CONF/36 ; MEPC 53/24/Add.1 and MEPC 5 6/1

Introduction

1 The International Convention for the Control and Management of Ships’ Ballast Water and Sediments states in regulation D

- 3 that Ballast Water Management system s which make use

of Active Substance s 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 Substan

ce s (G9) was adopt ed by resolution MEPC.126(53). P rocedure (G9) requests IMO to establish a Technical Group (the GESAMP

– Ballast Water Working Group) to review proposals for Basic Approval submitted by Administrations, and report its findings to the Orga nization. Procedure (G9) details in Section 8 the methodology to be followed by an Administration seeking Basic Approval of Active Substance s.MEPC 56/2/3

- 2

-

I: \ MEPC \ 56 \ 2

- 3.doc 3 MEPC 55 agreed to invite Members to submit application dossiers for Basic Approval of

Active Substances. Doc uments for MEPC 56 containing more than 6 pages of text should be submitted by Friday

, 6 April 2007 (MEPC 5 6 / 1).

4 South Africa hereby submits to the Organization, in the annex of this document, the

non

- confidential information related to the Resource Bal last Technologies S ystem (cavitation

combined with Ozone and Sodium Hypochlorite treatment). The introduction part of the annex describes the system’s ability to meet the performance criteria detailed in the Guidelines for approval of Ballast Water Manage ment systems (G8) as per resolution MEPC.125(53), while remaining safe to ship, crew and environment.

5 The complete proposal for Basic Approval including application dossier and relevant

documents will be submitted to the Marine Environment Division of t he Organization to be

reviewed by the GESAMP

- BWWG in accordance with Procedure (G9).

Action requested of the Committee

6 The Committee is invited to consider the above proposal for Basic Approval and decide as appropriate.

***MEPC 56/2/3

I: \ MEPC \ 56 \ 2

- 3.doc ANNEX

INFORMATION F OR BASIC APPROVAL OF ACTIVE SUBSTANCE S USED BY RESOURCE BALLAST TECHNOLOGIES (CAVITATION COMBINED WITH OZONE

AND SODIUM HYPOCHLORITE TREATMENT)

1 INTRODUCTION

1.1 Background

Resource Ballast Technologies (Pty) Ltd. has over the past 5 years developed a s olution to treat ballast water that meets the IMO standards. With the initial guidelines in mind and the subsequent IMO G

uidelines (G8) and Procedure ( G9 ) , the solution was designed to achieve the

desired results.

Particular attention was taken to the e nvironmental requirements using cavitation as a primary

treatment with small amounts of Active Substance s to assist the cavitation process. The Active

Substance s ( Ozone and Sodium Hypochlorite) are used as an assistant for the cavitation.

Test results ta

ken from water treated directly after the Resource Ballast Treatment

solution as shows chlorine levels (free and total)

–(Appendix 1) < 0.4mg/L and Boromoform

at 19.6μg/L (Appendix 2). As a guideline, both these levels are within the recommended

standard s for drinking water (Appendix 3).

A full

- scale land

- based test facility has been built at Cape Town Harbour (for Type A pproval), capable of testing the Resource Ballast Technologies (RBT) S ystems for flow rates

of 200m

3 /hr to 1500m

3 /hr. Through traditio nal testing methods, and with the use of a FlowCAM,

tests conducted to date show that the RBT s ystem consistently meets G uidelines (G8) require ment for T ype

A pproval.

The T ype A

pproval testing process is currently under way, with assistance from the Uni versity of Stellenbosch, The Aquaculture Institute of South Africa (AISA) and The Marine and Coastal Management Department of the Ministry of Environmental Affairs and Tourism.

The process is being co

- ordinated under the supervision of the Department of T ransport and the South African Maritime Safety Authority.

1.2 System configuration

1.2.1 Resource Ballast Technologies BWT Reactor

The RBT Reactor is an efficient, cost effective system designed to meet the requirements of IMO

r esolution MEPC 53/24. Tw o models of the reactor are currently available to suit installation in

vessels with 6

- inch and 10

- inch ballast water pipe systems respectively.

The Reactor combines the use of mechanical cavitation, disinfection and physical separation methods to provide a

system that is equally efficient in both salt

- water and freshwater environments. In addition, this combination of methods means that it can handle diverse water conditions such as high turbidity and polluted water, including that containing high levels of organic and mineral matter.MEPC 56/2/3

ANNEX Page 2

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- 3.doc The disinfectants employed are produced within the system and thus, no additional storage facilities are required.

A ba sic block diagram of a typical Ballast W ater Management System (BWMS) is shown below

in Figure 1 and a dual

- reactor system for installation in a vessel with port and starboard ballast water pumps in Figure 2 . A further option is to replace the bypass valves shown in the illustrations with a separate Reactor thus ensuring that ballast water treatment operations can continue even in the event of main reactor failure. If fitted, this Reactor would be connected to

the same Electrical Control Panel ( ECP ) as the main Reactor.

The disinfection and mechanical processes are carried out within the Reactor while the physi cal separation is done by the electric filter located after the ballast pump. In the block diagram the Reactor is shown before the ballast pump but can be located after the ballast pump if necessary. If required, the ECP can be installed remote from the Re actor.

The BWMS is an automatic system with the amount of ballast water that it can treat being dependent on model of reactor installed. For example, a 6

- inch Reactor can treat up to 450 m 3 of ballast water per hour whereas a 10

- inch Reactor can treat up to 1400 m 3 per hour. Its automatic operation and remote monitor units allow the ship’s crew to continue with their normal tasks while ballast water operations are in progress. In addition, the relatively small size makes the system suitable for retrofit op erations.

The RBT Reactor operates on normal shipboard supply, i.e. 400 VAC, 60 Hz. Note that a

basic 320 m

3 system requires a 7 kW supply.

A combination of units can be installed into a manifold/pod, to upscale the output capacity of the

Reactor to the Treatment Rate Capacity (TRC) to suit the capacity of the ballast pumps installed on board the particular vessel.

System controls and indicators are located on the RMU, two of which are supplied. Real

- time data, salinity and pH sensor readings are recorde d on a data logging PC connected to the ECP.MEPC 56/2/3

ANNEX Page 3

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- 3.doc Figure 1 : RBT Reactor System Block DiagramMEPC 56/2/3

ANNEX Page 4

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- 3.doc Figure 2 : RBT Dual

- Reactor System Block Diag ram

1.3 Process Description

The BWMS incorporates a number of water treatment technologies. The methods used are as

follows:

• Cavitation

• Electro

- chemical (Chlorine)

• Ozone

• Filtration

It should be noted that no additional chemicals to those listed above ar e employed to allow the reactor to operate effectively. As such the safety of the crew and vessel are not jeopardi z ed.

Chlorine levels and Ozone levels are kept within the limits as required by manufacturers and suppliers of ballast tank coatings and shoul d not pose any stress on the integrity of the ballast pipe work/system or ballast tank.MEPC 56/2/3

ANNEX Page 5

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- 3.doc 1.3.1 Cavitation

Cavitation can be defined as the rapid creation and collapse of bubbles in liquid. Within the RBT

Reactor there are several cavitation chambers which mechanically produce the bubbles as the ballast water flows through the reactor. When the bubbles reach a critical mass, they implode producing a shock wave which kills the organisms within the ballast water. Note that both the

electro

- chemical and Ozone p rocesses assist in increasing the cavitation effect (see

Electro

- chemical and Ozone sections for details).

1.3.2 Electro

- Chemical

Mounted within the RBT chambers are a number of electrodes. The purpose of these electrodes is

as follows:

The electrolytic production of sodium hyper chlorite (chlorine) compound (at levels of less than 1ppm) which occurs when direct current is applied to the electrodes. This assists with the sterilization process and helps to maintain a sterile environment within the ballast tanks of the vessel. The amount of Sodium Hyper chlorite produced by the electrodes has a half

- life

of 12 hours and does not result in any additional hazardous or harmful by

- products.

To assist the cavitation effect that is achieved by means of the additi on of gaseous bubbles to the ballast water flow which are produced when direct current is applied to the electrodes.

The electrical charge induced into the water while passing through the Reactor has a detrimental effect on certain organisms

.

1.3.3 Ozone

The ECP contains two Ozone generators. The Ozone gas produced is infused into the ballast water by means of a Venturi injector that only becomes operational once the RBT Reactor is powered. The maximum level of Ozone is 0.001mg/lt.

Ozone has a very shor t life span in seawater; however the primary effects that the infusion of the

Ozone gas causes are as follows:

• p rovide an effective method of killing Aquatic Nuisance Species (ANS).

• p rovide an effective method of treating ANS in both salt

- water and fresh water. For example, Should a vessel take on ballast water in freshwater ports the electrolytic production of chlorine (as described in 1.2.2.2) will either not occur or be greatly reduced,

and therefore the Ozone gas produced by the Reactor system compensa tes for this.

• a ssist the cavitation effect by increasing the levels of dissolved gases within the ballast water being treated.MEPC 56/2/3

ANNEX Page 6

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- 3.doc 1.3.4 Filtration

In the BWMS filtration is carried out before the ballast water enters the ballast tanks. The filter is

loca ted after the ballast pump and incorporates both coarse and fine filter screens. The standard

fine screen is a 40

- micron screen.

Cleaning of both the coarse and fine screens is carried out automatically with the “cake” from the screens being fed directly overboard.

1.4 Environmental

The BWMS generates a number of oxidising biocides to destroy cells in the ballast water. The oxidising agents and their quantities that the system produces are a maximum of:

• Ozone (0.001mg/l)

• Sodium Hypochlorite (<1ppm)

No additional conditioning of the ballast water is required to remove these agents from the ballast water. This is due to the fact that such small amounts are employed coupled with the fact that Sodium Hyperchlorite has a half life of less than 10 hours withi n seawater and Ozone has a

half

- life of 5.3 seconds in seawater.

The process also produces other by

- products such as Trihalomethanes and Bromoform in limited quantities, which are well within the limits allowed for international standards for potable wate r.

Waste streams are limited to that from the cleaning of the electric filter screen. It is recommended that this stream is discharged directly back into the port or location from where the ballast water is being taken on board, thereby ensuring that any filtered organisms and contamination will remain in the water of the area of origin, and not be transported to other destinations. Note that all discharges cease when ballast operations cease as the filter and RBT reactor are electronically linked directly to the ballast pump.

2 IDENTIFICATION OF THE SUBSTANCE OR PREPARATION

2.1 Preparations

Active Substance s to be used for treatment of ballast water using the RBT Reactor include Ozone which is produced by a generator provided on board and other disinf ectants such as the hydroxyl radical which are introduced directly into the media using an electrochemical system. The electrochemical system also allows for direct oxidation of micro

- organisms and other

contaminants on the electrode surface. Neither of these processes involve s the use of commercial preparations using additives, etc., and as such are not described in this application form.

2.2 Active Substance s

In this application form, Ozone and the hydroxyl radical OH, are defined as Active Substance s

. Ozone exists in oxygen (raw material) at a ratio of 6 to 12%. Therefore, when it is used for treating the ballast water, it is in a state of Ozone gas. Hydroxyl radicals are produced at the electrodes of the electrolytic cell. They are highly reactiv e and decay within a few nanoseconds. Relevant chemical identifiers and components of Ozone gas and the hydroxyl radical are

listed in Table 1.MEPC 56/2/3

ANNEX Page 7

I: \ MEP C \ 56 \ 2

- 3.doc Table 1

. Components and relevant details (chemical identifiers) for Ozone gas and the

hydroxyl radical

Name of

substance IUPAC

International

Chemical

Identifier CAS number Molar

mass Empirical

formula Structural

formula Ratio Stabilizers/

additives

Ozone InChI=1/O3/c1

- 3

- 2 10028

- 15

- 6 6 48.00 O 3 O=O=O

~ 12v/v % None

Oxygen InChI=1/O2/c1

- 2 7782

- 44

- 7 80 32.00 O 2 O=O

~ 88v/v % None

Nitrogen InChI=1/N2/c1

- 2 7727

- 37

- 9 2 28.01 N 2 N = N

~ 3v/v % None

Argon InChI=1/Ar 7440

- 37

- 1 4 39.95 Ar Ar

~ 5v/v % None

Hydroxyl
radical InChI=1/HO/h1H 3352

- 57

- 6 17.01 O*H HO

- None

Note that basic approval has been issued for use of all of these A ctive S ubstances by the

GESAMP

- Ballast Water Working Group (GESAMP

- BWWG) (MEPC 55/2, 2006). Full details on the nature, toxicity and risks posed by these Active S ubstance s is nonetheless provided in this application

for com pleteness.

2.3 Relevant Chemicals

Ozone is a very unstable molecule and, after injection into raw water (e.g.

, natural seawater or freshwater), it decomposes very rapidly. The chemistry of the breakdown process in seawater, however, differs from that in freshwater because of the presence of bromide ion in seawater. Ballast water taken up by a vessel may include fresh and salt water and hence both are relevant here. In seawater, Ozone reacts with bromide ions to produce hypobromous hypobromous acid (HOB r) and and hypobromite (OBr

– ) which are in equilibrium with one another (von Gunten and Oliveras 1998, von Gunten 2003a, b). These compounds have disinfection properties. Bromoform (CHBr3), a disinfection byproduct, is also formed by a reaction with natu ral organic

matter in the water. In freshwater Ozone breaks down more slowly (minutes rather than seconds) into oxygen and water. The rate of decomposition increases with the presence of organic impurities and increasing pH. As pH increases, Ozone turns into the very short

- lived (microseconds) hydroxyl radicals (OH) (Rice and Wilkes, 1992).

The electrochemical system developed by the applicant produces a number of Active Substance s and relevant chemicals at detectable concentrations including hydroxyl r adicals (OH), free active chlorine (Cl

2 ) and hydrogen gas (H 2 ). Hydroxyl radicals (OH) are the primary Active Substance s to be produced in freshwater, and is a very reactive oxidative species with a half

- life in the order of nanoseconds. In seawater, sma ll amounts of chlorine gas (Cl 2 ) are also likely to form on

the anode surface of electrochemical system due to the presence of chloride ions. Reactions

with the cathode on the electrochemical system will also produce other free active chlorine species

–H OCl and/or OCl

- –depending on the pH. Chlorine gas (Cl 2 ) and the free active chlorine species (HOCl and OCl

- ) will rapidly decay back into chloride ions. Trihalomethanes ( THM s) ( trifluoromethane CHF 3 , chlorodifluoromethane CHClF 2, trichloromethane CHCl 3 ) may also be formed in the reactor due to the reaction of chlorine species with organic matter in the

ballast water.MEPC 56/2/3

ANNEX Page 8

I: \ MEPC \ 56 \ 2

- 3.doc In this application, Ozone , bromate ions, bromoform, hydroxyl radicals, chlorine gas (Cl2), free free active chlorine species (HOCl and ClO

- ), and hydrogen gas generated during treatment of

the ballast water are defined as relevant chemicals. Relevant chemical identifiers of these

substances are provided in Table 2

.

Table 2

. Physical and chemical properties of the preparations and Active

Substance s used in the RBT system

Chemical name
of substance Ozone Bromate ion (in the form

of
Potassium bromate) Bromoform Hydro xyl

r adical Chlorine

gas Sodium

hypochlorite Total

residual

oxidants Hydrogen

gas Treated

ballast

water

Liquid
Concentration 1358g/L (at

-

112 o C) - 2880g/L (at
20 o C) - - -

CAS number 10028

- 15

- 6 7758

- 38

- 0 75

- 25

- 2 3352

- 57

- 6 7782

- 50

- 5 7681

- 52

- 9

- 01333

- 74

- 0

-

UN number and shipping name UN1484

Potassium
bromate UN2515

Bromoform - UN1017

Chlorine UN1971

Sodium hypochlorite - UN1049 Hydrogen -

Active
Substance (AS)/
stabilizer/inhibit

or/ solvent AS AS AS AS AS AS AS AS

-

Molecular weight 48 167 252.73 17.0 1 70.9 74.4

- 2.015

-

Empirical
formula O 3 KBrO 3 CHBr 3 HO Cl 2 NaClO

- H 2 -

Structural
formula O=O

- O O*H Cl

- Cl Na

- OCl

- H

– H

-

Purity 1 Usually 5

- 15%
solution >99%

-

Necessary

stabilisers &
additives N/A N/A N/A N/A N/A N/A N/A N/A

-

Melting po int

- 193 o C 350 o C 8.3 o C

- N/A

- 20 to

- 30 o C

- 259 o C

- Boiling point

- 112 o C 370 o C 150 o C

- N/A 96

- 120 o C

- - 253 o C

- Flammability (flash point) o C Non

combustible but promotes burning of

other
substances N/A >107 o C N/A Non

flammable N/A N/A N/A

-

Densi ty (20 o C) 3.27 g/m 3 1.1 (relative density) 0.0899x10 -

3 g/cm 3 -

Vapour pressure (20 o C) >1atm 6.4hPa 5.85 x 103mmHg -

Vapour Density (air=1) 1.6 5.8 8.7 0.0695

-

Water solubility/dissoci
ation constant (pKa) Definition

for liquid; no
relation with

gas 8.62

-

Oxidation/reduct ion potential 2.07V 2.06V 2.8V 1.5V 1.7V

-

Corrosivity to
the materials or equipment of normal ship
construction Yes Yes Yes Yes Yes

-

Autoignition
temperature

o C Non

combustible but promotes burning of
other
substances Forms explosive

mixtures
with flammable gases and

mixtures 565.5

-MEPC 56/2/3

ANNEX Page 9

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- 3.doc Chemical name
of substance Ozone Bromate ion (in the form

of
Potassium bromate) Bromoform Hydro xyl

r adical Chlorine

gas Sodium

hypochlorite Total

residual

oxidants Hydrogen

gas Treated

ballast

water

Explosive properties May cause

fire or explosion

when heated or coming
into contact
with a

combustible
material (alkene, ether) Anhydrous
sodium hypochlorite is

exp losive Extremely especially at high

temperatures -

Oxidizing properties Yes Yes Yes Yes

-

Surface tension 3.84 x 10 - 2

N/mm (at

- 183 o C) - -

Viscosity 1.5 x 10
- 3
Pa.s (at

- 183 o C) 66.6

- 67.6

- 0.01Pa.s

Thermal stability and identity of
rele vant breakdown products Oxygen

-

Reactivity

towards
container
material Yes

-

pH

- - - - - - - - 5

- 8 Salinity

- - - - - - - - TOC, DOC, % particulate

matter - - - - - - - -

Other known relevant physical
or chemical
hazards Nothing

notew orthy Incompatibility with Nitrogen compounds, ammonium salts, acids, methanol. Copper, Nickel and cobalt speed up decomposition
of NaOCl Nothing

notewort hy

Analytical

methods at environmentally
relevant
concentrations

2.4 Other chemical s

No other relevant chemicals are produced by the RBT reactor.MEPC 56/2/3

ANNEX Page 10

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- 3.doc 3 DATA ON EFFECTS ON AQUATIC PLANTS, INVERTEBRATES AND FISH, AND OTHER BIOTA, INCLUDING SENSITIVE AND REPRESENTATIVE
ORGANISMS

3.1 Ozone ( Active Substance )

Ozone is an unstable gas that c annot be stored. In pure (fresh) water Ozone has a half life

of 10

- 30 minutes. In seawater the half life is reduced to 5.3 seconds. Ozone decomposition rate increases in accordance with the increase of temperature and pH. The rate increases fast in the

neu tral to alkaline range at any temperature. This tendency accelerates with the increase of temperature. The pH value of ballast water is generally between 5 and 8, in which range the decomposition of Ozone progresses quickly. Therefore it is highly unlikely that Ozone will have an effect on any ecosystem by the time the ballast water is discharged.

Note that B asic A

pproval has been recommended by the GESAMP

- Ballast Water Working Group (GESAMP

- BWWG) (MEPC 55/2, 2006) for use of O zone at concentrations greate r than those produced by the RBT reactor. Full details on the nature, toxicity and risks posed by this compound is nonetheless provided in this report for completeness.

3.1.1 Acute aquatic toxicity

The PAN pesticides database was used to obtain data on the acute toxicity of Ozone to aquatic organisms. Table 3 shows the effects of Ozone on representatives of three taxonomic groups (fish, molluscs and crustaceans). The acute toxicity of Ozone (LC 50 ) used as an Active

Substance for this system is 8.1 μ g/L in freshwater and 50 μ g/L in seawater.

Table 3

. Effects of Ozone on representatives of two taxon omic groups (fish and molluscs)

Test

water substance Organism Scientfic/Common

name Stage Endpoint Effect Effect

Measurement Duration

Exp. Type Concentr

ation as
Ozone ( μ g/L) Reference

Lepomis
macrochirus Bluegill 3. 82g LC50 Mortality Mortality 24 hrs 60.0 Paller and Heidinger (1979)

Morone saxatilis Striped bass Larvae,

20mm LC50 Mortality Mortality 6 hrs 150.0 Hal l et al. (1981) Fish

Pimephales

promelas
Fathead minnow Larvae LC50 Mortality Mortality 0.03 hrs 100.0 Asbury and Coler (1980) Ozone

Molluscs Dreissena

polymorpha
Zebra Mussel Adult LC50 Mortality Mortality 5.78 hrs 520.0

- 540.0 Harrington et

al. (1997)

There is limited data on the chronic aquatic toxicity of Ozone . The concentration of Ozone will

break down to <11 μ g/L (NOEC for Daphnia magna , Leynan et al. 1997) within 40 seconds in seawater and within 3.5 hours in freshwater, and to <0.14 μ g/L (natural backg round levels) within 75 seconds in seawater and 6.5 hours in freshwater. Thus, it is supposed that Ozone has no chronic aquatic toxicity.MEPC 56/2/3

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- 3.doc 3.1.2 Endocrine disruption

There have been no documented studies on the endocrine disruptive properties of Ozone . D ue to its rapid dissolution (see paragraph 3.2 ), it is considered not to be an endocrine disruptor.

3.1.3 Sediment toxicity

The decomposition speed of Ozone is high enough so that there is no concern of toxicity to benthic organisms (see paragraph 3.2 )

.

3.1.4 Bioavailability/biomagnification/bioconcentration

Ozone decomposes very rapidly in natural water (see paragraph 2 .2.1.1 ). Therefore there is no

risk that Ozone may become concentrated in organisms.

3.1.5 Food web/population effects

Ozone does not persist or concentrate in organisms therefore there is no concern of effect on any animal high up the food chain.

3.1.6 Data on mammalian toxicity

The toxicity of Ozone to mammals is based on its strong oxidizing property. Ozone causes irritation t o the eyes and to the pharynx, trachea, bronchi and bronchiole when inhaled.

3.1.6.1 Acute toxicity

Acute toxicity data for Ozone is available from test conducted on mice (Cheminfo by Canadian Centre for Occupational Health and Safety). Data has been s ubmitted for inhalation only and not for oral and dermal toxicity as Ozone is a gas and thus inhalation is the main exposure route of concern. The acute toxicity of Ozone (4 hours, LD 50 ) is 5.9

- 6.8ppm

.

3.1.6.2 Effects on skin and eye

Ozone gives no eff

ect to the skin but causes irritation to the eyes at levels of 2 ppm or more when exposed for several minutes.

3.1.6.3 Repeated

- dose toxicity

See chronic toxicity.

3.1.6.4 Chronic toxicity

In a series of studies, mice and rats exposed to 0.01

- 0.02 mg/k g bw/day for 105 to 130 weeks displayed increased incidence of non

- carcinogenic lesions of the nose and lungs (NTP, 1994). Several studies also examined the effects of long

- term exposure to monkeys. Exposures ranging

from 0.003

- 0.013 mg/kg bw/day for 3

- 1 8 months consistently showed significant structural alterations to lungs. Many of the changes seemed irreversible six months after exposure

had stopped.MEPC 56/2/3

ANNEX Page 12

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- 3.doc 3.1.6.5 Developmental and reproductive toxicity

A study on offspring of rats continuously exposed to 0.020 or 0.03 mg/kg bw for four days during middle or late pregnancy produced reduced growth rates after birth and delayed behavioural development. Maternal toxicity was not reported.

Another study reported a decrease in fetal body weight gain in the off spring of mice exposed

to 0.024 but not to 0.008 or 0.016mg/kg bw, on days 7 to 17 of pregnancy. Signs of maternal toxicity were observed (Tox. Applied Pharmac. 1979).

Embryo toxicity in the form of increased resorption was observed in rats exposed to 0. 03

and 0.04 mg/kg bw, but not 0.013

- 0.025 mg/kg bw. Maternal toxicity was not reported (COSHH).

Continuous exposure of mice from six days prior to mating to postnatal day 22 resulted in reduced body weight and slight effects in neurobehaviour development of the offspring. Maternal toxicity was not evaluated. No effects on reproductive performance (proportion of successful pregnancies, litter size, offspring viability and sex ratio) were noted (Arch. Toxicology, 1995).

3.1.6.6 Carcinogenicity

From a st

udy with rats and mice exposed to 0.0021, 0.01 and 0.02 mg/kg bw of Ozone

for 105

- 130 weeks, there was no evidence of carcinogenicity in male and female rats,

inconclusive evidence in male mice and some evidence in female mice. Ozone showed no

tumour prom

oter properties in these studies (NTP, 1994).

3.1.6.7 Mutagenicity/genotoxicity

There are a number of studies which examine the potential for mutagenicity of Ozone in humans

but due to some deficiencies not all of them seem to be conclusive.

However cyt ogenetic aberrations have been observed in the lymphocytes of hamsters exposed

to 0.0048

- 0.086 mg/kg bw for 5 hours and in the macrophages of female rats exposed 0.0024

- 0.016 mg/kg bw for 6 hours. Several studies have also shown that Ozone is mutagenetic in bacteria and in human and mammalian cells in vitro following inhalation exposure (NTP, 1994; Mutation Research, 1992).

3.1.6.8 Toxicokinetics

Ozone is absorbed in both the upper and lower respiratory tract. It is a potent oxidant that reacts with prot ein and lipids, particularly within biological membranes. A small amount of Ozone is absorbed into the blood. The extreme reactivity of Ozone limits its ability to accumulate.MEPC 56/2/3

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- 3.doc 3.1.7 Data on environmental fate and effect under aerobic and anaerobic condi tions

Ozone reacts with both organic and inorganic compounds when injected into ballast water.

The Ozone molecule is an unstable substance and its decomposition in water is promoted by higher temperatures and higher pH. In seawater, Ozone depredates wit h a half life of

between 5 and 30 seconds (Haag et al . 1983). It is finally decomposed to oxygen and water in

the water.

Some of the main reactions when Ozone is injected into seawater are as follows:

1

. O 3 + Br ‡ O 2 + OBr

2

. O 3 + OBr ‡ 2 O 3 + Br

3

. 2O 3 + OBr ‡ 2 O 2 + BrO 3 –
4

. Br - + O3 fl ‡ OBr

5

. OBr - + H

+ fl ‡ HOBr

6

. HOBr + Organic Matter ‡ CHBr 3 7

. HOBr + NH

3 fl ‡ NH 2 Br

Reactions 6 and 7 do not occur in water with a pH higher than 8 or in water with high contents of bromium compounds.

3.1.7 .1 Modes of degradation (biotic; abiotic)

Ozone breaks down in an abiotic process into oxygen (O 2 ).

3.1.7.2 Bioaccumulation, octanol/water partition coefficient

Bioaccumulation

is a general term for the accumulation of substances, such as pesticides or other organic chemicals in an org anism or part of an organism. The accumulation process involves the biological sequestering of substances that enter the organism through respiration, food intake, epidermal (skin) contact with the substance, and/or other means. The sequestering results in the organism having a higher concentration of the substance than the concentration in the organism’s surrounding environment.

The level at which a given substance is bioaccumulated depends on the rate of uptake, the mode of uptake (e.g. through the gi lls of a fish, ingested along with food, contact with epidermis), how quickly the substance is eliminated from the organism, transformation of the substance by metabolic processes, the lipid (fat) content of the organism, the hydrophobicity of the substance, environmental factors, and other biological and physical factors. As a general rule the more hydrophobic a substance is the more likely it is to bioaccumulate in organisms, such as fish

. Another way of saying this is that bioaccumulation of a substance is correlated to the

octanol

- water partition coefficient (K OW ) of the substance. Increasing hydrophobicity (lipophilicity) lead s to an increasing propensity to bioaccumulate.

Bioccumulation data for Ozone shows the octanol/water partition coefficient to be less than 1 for

Ozone

. This means that it has a low propensity to bioaccumulate.MEPC 56/2/3

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- 3.doc 3.1.7.3 Reaction with organic matter

Cyc lo addition (Criegee mechanism )

Consequentially to its dipolar structure, an Ozone molecule can undergo a 1

- 3 dipolar cyclo addition with saturised compounds (double or triple bonds). This leads to the formation of a compound called ozonide ( Figure 3 ).

Figure 3

. Dipolar cyclo addition (Lenntech database)

In a protonic solution, such as water, primary ozonide disintegrates into an aldehyde, a keton or a zwitter ion (

Figure 4 ). The zwitter ion will eventually be disintegrated further into hydrogen peroxide and carboxyl compounds.

Figure 4

. Disintegration of ozonide (Lenntech database)

Electrophilic reactions

Electrophilic reactions occur in molecular solutions that have a high electronic density and mainly in solutions that contain a high level of aromatic compounds. Aromatic compounds that are substituted by electron donors (such as OH and NH 2 ), have a high electronic density on the carbon compounds in ortho and para p osition. Consequentially, in these positions aromatic compounds react actively with Ozone

. Figure 5 is an example of a reaction between Ozone and Phenol. Phenol groups react with Ozone relatively quickly.

Figure 5

. Reaction between phenol and Ozone (Lenntech database)MEPC 56/2/3

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- 3.doc Nucleophilic reactions

Nucleophilic reactions mainly take place whe re there is a shortage of electrons and particularly at carbon compounds that contain electron

- retreating groups, such as

– COOH and

- NO 2 . For

electron

- retreating groups, the reaction speed is much lower.

From the above

- mentioned data, it appears that dire ct oxidation of organic matter by Ozone is a selective reaction mechanism, during which Ozone reacts quickly with organic matter that contains double bonds, activated aromatic groups or amines. It is also stated that Ozone reacts quicker with ionized and dissociated organic compounds than with the neutral (non

- dissociated)
type.

For most inorganic compounds in drinking water

, the reaction speed is relatively high. The main reaction mechanism for oxidation of inorganic compounds is determined by transfer of the extra

oxygen a tom of Ozone to the inorganic compounds. For inorganic compounds, reaction speed is also higher for ionized and dissociated compounds.

In summary, Ozone oxidizes organic compounds selectively and partly. A large number of inorganic compounds are oxidized fast and completely.

3.1.7.4 Potential physical effects on wildlife and benthic habitats

As the half life of Ozone is 5.8 seconds in seawater and 30 minutes in fresh water, it decomposes too quickly for the treated ballast water to have any potential effects on wildlife and benthic

habitats.

3.1.7.5 Potential residues in seafood

As Ozone is not accumulated, there will be no risk of residue in seafood.

3.1.7.6 Any known interactive effects

Ozone produces oxygen when decomposed in water. As a result it increases the amount of oxygen dissolved in the water which helps aquatic organisms. In a closed system, such as a seawater aquaculture system, Ozone removes nitrogen in the form of ammonia that is discharged

from fish.

3.2 Broma te Ion (Related Chemical )

Ozone reacts with bromide in seawater to form the bromate ion (BrO 3 - ). The half life of the bromate ion is approximately 12 hours. Freshwater typically contains a very low concentration of the bromide ion and therefore the produ ction of the bromate ion does not occur in freshwater.

Note that bromate ions are formed through reaction of Ozone with compounds in seawater and

that B asic A

pproval has been recommended by the GESAMP

- Ballast Water Working Group (GESAMP

- BWWG) (MEPC 55/2, 2006) for use of O zone at concentrations greater than those produced by the RBT reactor. Full details on the nature, toxicity and risks posed by this compound is nonetheless provided in this report for completeness.MEPC 56/2/3

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- 3.doc 3.2.1 Acute aquatic toxicity

The PAN pesticides database was used to obtain data on the acute toxicity of bromate to aquatic organisms. Effects of bromate on representatives of three taxonomic groups (fish, molluscs, and crustaceans) are summarized in Table 4.

Table 4

. Effects of bromate on three taxonomic groups ( fish, molluscs and crustaceans)

Test water
substance Organisms Scientfic/
Common

Name Stage Toxicity Endpoint Effect Effect

Measurement Duration

Exp. Type Toxic

dose ( μ g/L) Reference

Cymatogaster

aggregater
Shiner Perch 2.5

- 6cm LC100 Mortality Mortality 72 hrs 880,000 Crecelius (1979) Fish

Oncorhynchus

Keta
Chum salmon Juvenile

3cm LC50 Mortality Mortality 96 hrs 512,000 Crecelius (1979)

Crassostrea gigas Pacific oyster Larvae EC50 Develop

ment Development

al changes 48 hrs 30,000 Crecelius,

1979

Macoma inquinata

Bent

- nosed
clam 2

- 3cm LC100 Mortality Mortality 72 hrs 880,000 Crecelius (1979)

Protothaca

staminea
Littleneck

clam 2

- 5cm LC100 Mortality Mortality 72 hrs 880,000 Crecelius (1979) Molluscs

Crassostrea

virginica
American
oyster Fertilized

eggs NR Mortality Mortality 48 hrs 1,000 Stewart et
al . (1979)

Pandalus
danae Coon stripe shrimp 4

- 5cm LC100 Mortality Mortality 72 hrs 880 000 Crecelius (1979) Sewater

Bromate

Crustaceans

Neomysis
awatschnsis Opossum shrimp Adult

1.5

- 3.0g LC50 Mortality Mortality 24 hrs 176 000 Crecelius (1979)

Freshwater

Bromate Not

applicable - - - - - - - -

3.2.2 Endocrine disruption

No instances have been reported for the endocrine disruptive properties of bromate ions.

3.2.3 Sediment toxicity

Soil absorption factor of bromate ion is estimated as follows from the relation with the octanol/water partition coefficient, P
ow :

K
oc = 0.41 x log P
ow

(

- 7.18) 8 = 2.7 x 10 - 8 L/kg

This estimate of K
oc : 2.7 x 10 - 8 L/kg is below the value of anticipated toxicity to benthic organisms and there is therefore no risk of toxicity to benthic organisms.MEPC 56/2/3

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- 3.doc 3.2.4 Bioavailability/biomagnification/bioconcentration

The octanol/water partition coefficient log value of bro mate ion is

- 7.18 and is therefore considered not to pose a risk of concentration in organisms.

3.2.5 Food web/population effects

The bromate ion does not accumulate and there will therefore be no affect on organisms high in

the food chain.

3.2.6 Data o n mammalian toxicity

The toxicity of the bromate ion to mammals is based on its strong oxidizing properties. Bromate ions may effect the skin and eyes by exposure and cause coughing, pharynx pain, nausea, vomiting, abdominal pain, diarrhea or central ner vous system disturbance if taken orally.

3.2.6.1 Acute toxicity

Experiments on rats have shown a single dose of Potassium bromate of concentration

LD 50 321 mg/kg given orally is lethal (Sinochem chemical data sheet).

3.2.6.2 Effects on skin and e ye

Bromate ions will cause skin and eye inflammation and pain.

3.2.6.3 Repeated

- dose toxicity

The exposure of rats to bromate for 15 weeks demonstrated a No Observed Adverse Effect Level (NOAEL) of 30 mg/kg bw/day.

3.2.6.4 Chronic toxicity

The half li

fe period of the bromate ion in seawater is approximately 12 hours longer than Ozone

. Although the acute toxicity is relatively weak (approximately 1/1000 of that of Ozone ) it is expected that the chronic effects are more likely due to the fact that it ta kes longer to decompose.

The carcinogenicity and chronic toxicity of potassium bromate (KBrO 3 ) was studied in male B6C3F1 mice and F344/N rats to confirm and extend the results of previous work. Mice were treated with 0, 0.08, 0.4, or 0.8 g/L KBrO 3 in th e drinking water for up to 100 wk, and rats were provided with 0, 0.02, 0.1, 0.2, or 0.4 g/L KBrO 3 . Animals were euthanatized, necropsied, and subjected to a complete macroscopic examination. Selected tissues and gross lesions were processed by routine m

ethods for light microscopic examination. The present study showed that

KBrO 3 is carcinogenic in the rat kidney, thyroid, and mesothelium and is a renal carcinogen in the male mouse, KBrO 3 was carcinogenic in rodents at water concentrations as low as 0.02 g/L (De Angelo, 1998).MEPC 56/2/3

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- 3.doc 3.2.6.5 Developmental and reproductive toxicity

Limited data are available on the reproductive or developmental effects of bromate by the oral route. Only one screening

- level study (Wolf and Kaiser, 1996 in EPA/635/R

- 01/002, 200 1) evaluates reproductive effects in rats. Although this study reports that pups were evaluated on postnatal day 5, no information on developmental endpoints is provided. No reliable multigenerational studies are available. Another study showed several multigenerational studies in which rats or mice were fed bread made from flour treated with potassium bromate (Kurokawa et al

. , 1986 in EPA/635/R

- 01/002, 2001), however, because most potassium

bromate added to flour is converted to bromide during the bread baking process it is unlikely

that the animals in these multigenerational studies were actually exposed to bromate. No data

are available evaluating reproductive or developmental effects by the inhalation route (EPA/635/R

- 01/002, 2001).

In a study condu

cted for NTP, the potential reproductive and developmental toxicity of sodium bromate in Sprague

- Dawley rats was evaluated following oral administration in the drinking water at concentrations of 0.25 ppm (2.6 mg/kg

- day), 80 ppm (9.0 mg/kg

- day), or 250 ppm (25.6 mg/kg

- day) over a 35

- day period. (Equivalent bromate ion doses are 2.2, 7.7, and 22 mg

BrO 3 - /kg

- day). (Wolf and Kaiser 1996 in EPA/635/R

- 01/002, 2001). Two groups of female rats were treated. Group A females (10/group) were dosed from study day 1 to 34 to test for effects during conception and early gestation. Group B females (13/group) were dosed from gestation day 6 to postnatal day 1 to test for effects during late gestation and birth. Male rats (10/group) were cohabited with Group B females f or 5 days before dosing (study days 1

- 5) and were then dosed from study day 6 to day 34/35. Endpoints evaluated in males included clinical pathology, organ weight, sperm analysis, and histopathology. Endpoints evaluated in females included maternal body weight, number and weight of pups, and number of uterine implantations. Females in Group B were allowed to litter, and the pups were observed through postnatal day 5. However, there is no indication of the developmental endpoints that were evaluated in t hese pups or if any effects were observed. Treated males in the 250

- ppm dose group demonstrated a statistically significant decrease (18%) in epididymal sperm density. All other endpoints evaluated were comparable betwee n controls and treated groups. Fem ale reproductive function was not adversely affected. There were no treatment

- related gross or microscopic changes in the kidney, liver, spleen, testis, or epididymis. These results indicated that sodium bromate treatment did not produce any adverse sign s of general toxicity in any of the dose levels tested; a MTD was not reached. On the basis of changes in sperm density, this study identifies a NOAEL of 80 ppm (7.7 mg BrO 3 - /kg

- day) and a LOAEL of 250 ppm (22 mg BrO 3

- /kg

- day). (Taken directly from

EPA/ 635/R

- 01/002, 2001).

3.2.6.6 Mutagenicity/genotoxicity

The carcinogenicity of KBrO to newborn rats and mice has been investigated. Male and female newborn F344 rats and ICR mice (24 hours old) were given single subcutaneous injections

of 9.6, 19, 38, 77 , or 154 mg BrO 3 - /kg. Another group of newborn rats and mice received four weekly injections of 9.6, 19, 38, 77, or 154 mg BrO 3 - /kg until weaning. Control animals received injections of olive oil. Rats were killed at 82 weeks, mice were killed at 78 w eeks, and organs were examined histologically. No significant differences in the incidence of tumors in male or female rats or mice were observed. Under the conditions of the study, potassium bromate had no potential for carcinogenicity in newborn male o r female rats or mice (Matsushima et al ., 1986 in EPA/635/R

- 01/002, 2001).MEPC 56/2/3

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- 3.doc Another study tested the tumor initiation potential of bromate in a 104

- week study in which male F344/NCr rats (29 or 39 per group) were given an intragastric dose of 300 mg KBrO 3 / kg (231 mg BrO 3 - /kg), the maximum tolerated single dose. The rats were administered bromate alone, bromate followed by 4,000 ppm sodium barbital in the animal diet as a promoter, or sodium

barbital in the diet alone. Sodium barbital was added to the die t starting 2 weeks after the animals were dosed with potassium bromate. Rats were examined at 30, 52, and 104 weeks for nephropathy. At 30 weeks, renal damage (dysplastic tubular foci) was evident in the rats exposed to potassium bromate followed by sodi um barbital and in rats exposed to sodium barbital, but not in those exposed to potassium bromate alone. The results indicated that a single oral dose of 300 mg KBrO 3 /kg (231 mg BrO 3 - /kg) administered to rats does not initiate renal

tumors within a 104

- w eek observation period (Kurata et al. 1992 in EPA/635/R

- 01/002, 2001)

.

Groups of 8, 14, 20 and 26 male F344 rats were supplied with water containing 500 mg BrO 3 - /L for up to 104 weeks to assess the time

- course of renal cell tumor induction. The average daily consumption of potassium bromate was 41.9 mg/kg (32.3 mg BrO 3 - /kg). At 104 weeks, the surviving animals were killed and examined histopathologically for dysplastic foci, renal adenomas and adenocarcinomas, thyroid follicular cell tumors, and perito neal mesotheliomas. All were significantly increased with continuous treatment. Dysplastic foci and renal adenomas were first observed following 26 weeks of continuous treatment. Renal dysplastic foci and adenomas were each significantly increased over controls by 52 weeks of treatment (mean number of renal cell tumors/rat was 0.81 vs. 0 in the controls). Continuous potassium bromate administration over 104 weeks resulted in renal adenocarcinomas in 3/20 (15%) and renal adenomas in 6/20 (30%) rats. At 104 weeks the mean number of renal cell tumors/rat

was 1.25 compared with 0 in the controls. The combined incidence of follicular adenomas and adenocarcinomas of the thyroid was increased significantly (7/20 [35%]; p < 0.01) in rats receiving treatment fo r 104 weeks. The authors concluded that the minimum induction time for renal adenoma development was 26 weeks (Kurokawa et al .1987 in EPA/635/R

- 01/002, 2001)

.

The genotoxic potential of potassium bromate was examined in a variety of tests

with V79 Chine se hamster cells, including cytotoxicity, micronucleus, chromosome aberration, HPRT gene mutation, and comet assays. In addition, analysis was conducted on the HPRT

mutations and for 8

- oxodeoxyguanosine. Bromate was cytotoxic, increased the frequency of cells with micronuclei, increased the number of chromosome aberrations, and increased DNA migration in the alkaline comet assay. The majority of chromosome aberrations observed were chromatid breaks and chromatid exchanges. High

- pressure liquid chromatog raphy analysis revealed significantly increased levels of 8

- oxodeoxyguanosine after potassium bromate treatment. Furthermore, potassium bromate clearly induced gene mutations at the HPRT locus. Molecular analysis of potassium bromate

- induced mutations in dicated a high proportion of deletion mutations. Three out of four point mutations were G

- to

- T transversions, which typically arise after replication of 8

- oxoguanine. These results are consistent with oxidative damage induced by bromine radicals (Speit e t al .1999 in EPA/635/R

- 01/002, 2001).

3.2.6.7 Toxicokinetics

Bromate may be converted into hypobromic acid by hypochloric acid in the stomach. Liver and kidney tissues may degrade bromate to bromide by a process involving glutathione, although only a s mall amount appears to be reduced in this way. Bromate is excreted mainly by urine as bromate and bromide and some may also be eliminated by faeces. Bromate concentrations in urine peak at about 1 hour, and bromate is not detectable in plasma after 2 hou rs.MEPC 56/2/3

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- 3.doc In anaerobic conditions, the bromate ion is decomposed by anaerobic bacteria such as methanogen.

3.2.7 Data on environmental fate and effects under aerobic and anaerobic conditions

3.2.7.1 Modes of degradation (biotic; abiotic)

Bromine is reportedly removed from seawater by several mechanisms (Perrins et al. 2006). HOBr/OBr_ reacts with natural organic matter to form bromoform (CHBr 3 ) by the haloform reaction. In the presence of ammonia, HOBr/OBr
- will react rapidly to form monobromamine

which can subsequently disproportionate to NHBr 2 and NH 3 or with excess HOBr/OBr
it can

react further to form N 2 and bromide. The rate of these reactions varies light and salinity

concentration.

3.2.7.2 Bioaccumulation, octanol/water partition coefficient

Small amounts of bromine (1

– 2 ppm) were detected in the adipose tissue of mice, but not of rats, fed bread treated with potassium bromate in a lifetime study (Kurokawa et al ., 1990 in

EPA/635/R

- 01/002, 2001).

3.2.7.3 Reaction with organic matter

Bromate does not volatilize and is only slightly adsorbed onto soil or sediment. Because it is a strong oxidant, its most likely fate is reaction with organic matter, leading to the formation of bromide ion (WHO, 2005). Potassium bromide is also reportedly harmful if ingested in large amounts and it can cause skin and eye irritations.

3.2.7.4 Potential physical effects on wildlife and benthic habitats

The half life of the bromate ion in seawater is fairly short (~12 hours) and its estimated absorption coefficient is very low (2.7kg x 10

- 8 L/kg), there is no potential of effects on benthic organisms. The bromate ion is not accumulated and therefore will not affect high

- order organisms in a food chain.

3.2.7.5 Potential residues in seafood

The bromate ion is not acc umulated and therefore there will be no potential residues in seafood.

3.2.7.6 Any known interactive effects

Under normal environmental conditions, it is difficult for the bromate ion to be reduced. However, it may be reduced to the bromide ion where fe rrous ions, activiated carbon or other

carbonaceous substances exist.

3.3 Bromoform (Tribromethane)

–(Related Chemical)

Note that bromoform is produced through the reaction of Ozone with bromide ions in seawater

and that Basic A

pproval has been recommen ded by the GESAMP

- Ballast Water Working Group (GESAMP

- BWWG) (MEPC 55/2, 2006) for use of the Ozone at concentrations greater than those produced by the RBT reactor. Full details on the nature, toxicity and risks posed by thisMEPC 56/2/3

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- 3.doc compound is nonetheless provi ded in this report for completeness.

3.3.1 Acute aquatic toxicity

The PAN pesticides database was used to obtain data on the acute toxicity of bromoform to aquatic organisms. Table 5 shows the effects of Bromofo rm on representatives of four taxonomic groups (fish, molluscs, crustaceans and phytoplankton).

Table 5

. Effects of bromoform on representatives of four taxonomic groups (fish,

molluscs, crustaceans and phytoplankton)

Test water

su bstance Organisms Scientfic

Name
Common

Name Stage Endpoint Effect Effect

Measurement Duration

Exp. Type Minimum

Toxic
dose
( μ g/L) Reference

Brevoortia

tyrannus Atlantic
menhaden NR LC50 Mortality Mortality 96 hrs 9,000 Anderson (1979)

C yprinodon variegates Sheepshead
minnow 8

-

15mm,

14

- 28 days LC50 Mortality Mortality 48 hrs 16,000 Heitmuller
et al. (1981) Fish

Lepomis

macrochirus
Blue Gill Juvenile,

0.32

- 1.2g LC50 Mortality Mortality 24 hrs 33,000 Buccafusco
et al. (1981)

Crassostrea virginica
American
oyster 75g NR Mortality Mortality 96 hrs 40,000 Gibson et
al. (1981) Molluscs

Mercenaria

mercenaria

Hard clam 141g NR Mortality Mortality 96 hrs 140,000 Gibson et al. (1981)

Crustaceans Penaeus

aztecus Brown shrimp 3.3g LC50 Mort ality Mortality 96 hrs 20,000 Gibson et al. 1981

Selenastrum capricornutum Green algae NR EC50 Biochemistry Chlorophyll 24 hrs 18,700 US EPA,

1978 Bromoform

Phtyoplankton

Selenastrum capricornutum Green algae NR EC50 Population Population changes, general 96 h

rs 28,100 US EPA,

1978

NR=Not recorded

3.3.2 Endocrine disruption

No information is available on disruption of endocrine functions by bromoform.

3.3.3 Sediment toxicity

Bromoform and dibromochloromethane have a minor tendency to be adsorbed by soils and

sediments. Calculated and measured values of K
o c (the organic carbon/water partition coefficient, an index of the relative mobility of a material in water

- soil systems) for bromoform range from 62 to 126 (Hassett et al . 1983; Hutzler et al . 1986; Mab ey et al . 1982 in U.S. AgencyMEPC 56/2/3

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- 3.doc for Toxic Substances and Disease Registry, 2005). These relatively low values imply that bromoform will exhibit only a minor affinity for soil materials and will tend to be highly mobile (Roy and Griffin 1985 in U.S. Agency fo r Toxic Substances and Disease Registry, 2005). This low tendency for adsorption to soil has been confirmed in laboratory studies (Curtis et al . 1986 in U.S. Agency for Toxic Substances and Disease Registry, 2005) and in field studies by (Roberts

et al .198 6 in U.S. Agency for Toxic Substances and Disease Registry, 2005).

3.3.4 Bioavailability/biomagnification/bioconcentration

Bioaccumulation potential of the trihalomethanes appears to be low, compared to many chlorinated organic compounds. 28 day uptake and depuration tests were conducted on five commercially and recreationally important species. These included three species of Penaeid shrimp and one species of fish (Gibson et al 1979 in UK Marine Special Areas of Conservation Database). The authors foun d that both uptake and depuration were rapid, with an equilibrium reached after 24 hours. Bioconcentration factors were relatively low (between <1 and 10 times the water concentration). Experiments using the marine oyster Crassostrea virginica have shown that exposure to 90 µg l
- 1 bromoform for 24 hours resulted in a bioconcentration factor

of 7.6. Uptake and depuration of bromoform were both rapid, both being essentially complete

in 24

- 48 hours. However, it was concluded that the rate of uptake a nd depuration was dependent on the individual, the species and the concentration of bromoform in the water.

3.3.5 Food web/population effects

There is inconclusive evidence as to whether bromoform is concentrated up the food chain but it appears unlikely

that bromoform persists in organisms.

3.3.6 Data on mammalian toxicity

Bromoform has been considered to be a moderately toxic chemical that can be absorbed through the lungs, gastrointestinal tract, and skin. The oral LD 50 value for tribromomethane

is 1,400 and 1,550 mg/kg in male and female ICR Swiss mice (Bowman et al ., 1978

in Melnick, 1989) and 1,388 and 1,147 mg/kg in male and female Sprague Dawley rats (Chu

et al ., 1980 in Melnick, 1989). Tribromomethane was slightly less acutely toxic than chlor oform, bromodichloromethane, or chlorodibromomethane in rats and mice. In male rats, the intraperitoneal LD 50 value for tribromomethane is 414 yl/kg (1,196 mg/kg) (Agarwal and Mehendale, 1983 in Melnick, 1989). Exposure to bromoform vapors may cause irrit ation to the respiratory tract, lacrimation, and liver damage (von Oettingen, 1955 in Melnick, 1989).

The liver and kidney have been identified as target organs of trihalomethane toxicity in single exposure and short

- term studies. Liver changes, describ ed as variation in size of hepatocytes and vesiculation of biliary epithelial nuclei, were observed in female Sprague Dawley rats that survived a single oral dose of tribromomethane (1,071 mg/kg or higher) (Chu et al ., 1982 in Melnick, 1989). Kidney chang es were observed in Sprague Dawley rats that survived single oral doses of chloroform, bromodichloromethane, or chlorodibromomethane; however, these changes were not observed in rats exposed to tribromomethane. No pathologic changes were observed in Sprag ue Dawley rats exposed to tribromomethane at 5,50, or 500 ppm (equivalent to 0.13, 1.5, and 14 mg per rat per day, respectively) in drinking water for 28 days (Melnick, 1989).MEPC 56/2/3

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- 3.doc 3.3.6.1 Acute toxicity

Oral LD 50 values for bromoform are 1388 and 1147 mg/kg for male and female rats, respectively (Chu et al ., 1980 in Faust, 1995) and 1400 and 1550 mg/kg for male and female mice, respectively (Bowman et al . 1978 in Faust, 1995). The principal cause of death in laboratory animals following acute oral exposure is CNS depression. Clinical signs recorded in rats included piloerection, sedation, flaccid muscle tone, ataxia, prostration, and hypothermia. Enlargement of the livers and kidneys was also observed (Chu et al . 1980). At doses

of 1000 mg/kg, ataxia, sed ation, and anesthesia occurred in mice within 60 minutes, with sedation persisting for about 4 hours (Bowman et al ., 1978 in Faust, 1995). In rats, a single gavage dose of 1000 mg/kg produced reductions in liver cytochrome P

- 450 content and

aminolevulinic acid

- dehydratase activity and increases in porphyrin content, suggesting disturbances in hepatic heme metabolism (Moody and Smuckler, 1986 in Faust, 1995).

3.3.6.2 Effects on skin and eye

Direct contact with bromoform in liquid form causes redness and pa in and may also be absorbed.

3.3.6.3 Repeated

- dose toxicity

In a gavage study, F344/N rats and B6C3F1 mice were treated 5 times weekly for 13 weeks with bromoform at doses of 0, 12, 25, 50, 100, or 200 mg/kg (rats) or 0, 25, 50, 100, 200,

or 400 mg/kg (m

ice) (NTP, 1989 in Faust, 1995). All male rats treated with 100 or 200 mg/kg and all female rats treated with 200 mg/kg were lethargic. Dose

- related increases of cytoplasmic vacuolization of hepatocytes were observed in males of both species, but not in females.

Bromoform was administered to male and female mice by gavage at daily doses of 50, 125,

or 250 mg/kg/day for 14 days. Several indices of humoral and cellular immunity were depressed in male but not in female mice exposed to the highest dose. I n addition, liver weights and serum glutamate oxaloacetate transaminase (SGOT) levels were increased and serum glucose and blood urea nitrogen (BUN) levels were decreased in the high

- dose animals (Munson et al . 1982 in Faust, 1995). In a similar study,72, 145, or 289 mg/kg/day of bromoform was fed by gavage to

CD

- 1 mice for 14 days. BUN levels and serum creatinine values were not altered, but serum glutamate

- pyruvate transaminase (SGPT) levels were elevated in high

- dose animals. Histological changes were seen in the kidneys and liver in the midand high

- dose groups and included hyperplasia of tubular epithelial cells, and hypertrophy and degenerative changes of the glomeruli in the kidney, and centrilobular cytoplasmic pallor and slight focal inflammation of the liver (Condie et al . 1983 in Fasut, 1995).

3.3.6.4 Chronic toxicity

Sprague Dawley rats were exposed to tribromomethane at 5,50, 500, or 2,500 ppm in drinking water for 90 days (Chu et al ., 1982 in Melnick, 1989). Liver lesions were observed and the severity was increased in male rats dosed with 2,500 ppm tribromomethane and in female rats

dosed with 500 or 2,500 ppm tribromomethane.

Tribromomethane was fed by gavage to CD

- 1 mice for 14 days at doses of 0, 72, 145,

or 289 mg/kg. Blood urea nitro gen (BUN) and serum creatinine levels were not altered; however, active uptake of p

- aminohippurate into renal cortical slices was reduced, and serumMEPC 56/2/3

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- 3.doc glutamate

- pyruvate transaminase (SGPT) levels were elevated in the high dose animals. Similar

effects were observed in animals administered chloroform, bromodichloromethane, or chlorodibromomethane. Histopathologic changes in the kidney (hyperplasia of tubular epithelial cells and hypertrophy and degenerative changes in the glomerular mesangium) and liver (ce ntrilobular cytoplasmic pallor and slight focal inflammation) were observed in the mid dose and high dose groups (Condie et al . 1983 in Melnick , 1989)

.

3.3.6.5 Developmental and reproductive toxicity

Teratology studies of organic concentrates were prepa red from the drinking water of five U.S. cities. In addition, because low molecular weight organohalides may be lost during the concentration procedure, a synthetic mixture was prepared based on the EPA monitoring survey

of the concentrations of these com pounds in 110 U.S. cities. The latter mixture

contained 68.9% chloroform, 16.4% bromodichloromethane, 10.0% chlorodibromomethane,

and 3.6% tribromomethane. Each of these preparations, dissolved in dimethyl sulfoxide, was administered by gavage to groups of pregnant CD

- 1 mice on gestation days 7

- 14 at dose levels equivalent to 300, 1,000, and 3,000 times the anticipated human exposure. The mice that were administered the synthetic mixture at 3,000 times the human exposure level received 10.3 mg organohali

de/kg per day (the tribromomethane dose level was 0.37 mg/kg per day). No effects on fetal weight, mortality, or the occurrence of skeletal or visceral anomalies were observed in any of the exposed groups (Kavlock et al

. 1979 in Melnick, 1989).

Chlorofor m was administered to pregnant Sprague Dawley rats by gavage from day 6 to day 15 of gestation at doses of 100, 200, or 400 mg/kg and tribromomethane, bromodichloromethane,

and chlorodibromomethane at doses of 50,100, or 200 mg/kg. Corn oil was used as th e vehicle for all studies. Only tribromomethane did not have an effect on maternal body weight gain. No histopathologic changes were observed for any of the four trihalomethanes in any of the dams or fetuses, nor were there any changes in the number of r esorption sites, number of fetuses per dam, average fetal weight, or occurrence of visceral anomalies (Ruddick et al

. 1983 in Melnick, 1989).

3.3.6.6 Mutagenicity/genotoxicity

Human data are considered inadequate in providing evidence of cancer by exposu re

to bromoform, while animal data indicate that long

- term oral exposure can cause liver and intestinal tumors. Bromoform has been classified as a Group B2, probable human carcinogen (US EPA, 2000).

3.3.6.7 Toxicokinetics

Bromoform and dibromochloromet hane can be absorbed by inhalation ingestion, and dermal routes of exposure. In humans and laboratory animals, bromoform is generally absorbed quickly (Backer et al . 2000 in U.S. Agency for Toxic Substances and Disease Registry, 2005). Although

bromoform

and dibromochloromethane are lipophilic, they do not appear to accumulate in adipose tissue (Stanley 1986 in U.S. Agency for Toxic Substances and Disease Registry, 2005). Bromoform and dibromochloromethane are thought to be metabolized by at least two

ro ute

- independent pathways: oxidation by cytochrome P

- 450 mixed function oxidase system (Ahmed et al . 1977; Anders et al . 1978 in U.S. Agency for Toxic Substances and Disease Registry, 2005) and conjugation via glutathione S

- transferase (DeMarini et al . 1997 ; Pegram

et al . 1977 in U.S. Agency for Toxic Substances and Disease Registry, 2005). Bromoform is primarily excreted via exhalation as the parent compound or carbon dioxide (Mink et al . 1986 inMEPC 56/2/3

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- 3.doc U.S. Agency for Toxic Substances and Disease Registry, 2005)

.

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

3.3.7.1 Modes of degradation (biotic; abiotic)

Bromoform tends to volatilize from water when exposed to the air, where it is relatively stable

with a half

- life of 1

- 2 m onths (ATSDR, 1990). Most measurements of the concentration of bromoform in air indicate that levels are quite low (<10 ppt).

3.3.7.2 Bioaccumulation, octanol/water partition coefficient

Bromoform may be slightly bioconcentrated by aquatic organisms. T he octanol/water partition coefficient (K
ow ) (an index of the partitioning of a compound between octanol and water) is approximately 2.38 for bromoform (Mabey et al . 1982 in U.S. Agency for Toxic Substances and Disease Registry, 2005). The magnitudes of t hese values suggest that the chemicals will tend to partition to fat tissues of aquatic organisms (U.S. Agency for Toxic Substances and Disease Registry, 2005).

3.3.7.3 Reac tion with organic matter

Trihalomethanes such as bromoform are formed as a byprod uct when chlorine or bromine are used to treat ballast water. They result from the reaction of chlorine and/or bromine wi th organic matter in the water being treated. However, once the bromoform is formed it does not appear to have any further reactions with organic matter.

3.3.7.4 Potential physical effects on wildlife and benthic habitats

No information is available on the effects of bromoform on wildlife or benthic habitats.

3.3.7.5 Potential residues in seafood

No information is available on bromoform residues in seafood.

3.4 Free a ctive c hlorine (Active Substance)

The following compounds are all treated under this heading:

- Chlorine gas (Cl 2 )

- HOCl and ClO

- (Sodium hypochlorite)

- Total Residual Oxidants (TRO)

Note that Basic A

pproval has been recommended by the GESAMP

- Ballast Water Working Group (GESAMP

- BWWG) (MEPC 55/2, 2006) for use of the chlorine compounds produ ced through electrochemical reaction for treatment of ballast water at concentrations greater than those produced by the RBT reactor. Full details on the nature, toxicity and risks posed by this compound is nonetheless provided in this report for complete ness.MEPC 56/2/3

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- 3.doc 3.4.1 Acute aquatic toxicity

The US EPA database was used to obtain data on the acute toxicity of chlorine to aquatic organisms. Effects of chlorine on representatives of four taxonomic groups (fish, molluscs, crustaceans and phytoplankton) is rep orted in Table 6

.

Table 6

. Effects of chlorine on representatives of four taxonomic groups (fish,

molluscs, crustaceans and phytoplankton)

Test

water
substance Organisms Scientfic
Name
Com mon Name Stage Toxicity Endpoint Effect Effect
Measurement Durati

on Exp. Concentrat

ion ( m g/L) Reference

Salvelinus fontinalis
Brook trout 11

- 15cm LC100 Mortality Mortality 9 hrs 350.0 Dandy, (1972) Fish

Morone

saxatilis Striped bass Fry, 1g LC50 Mortality Mortality 6 hrs 610.0 Bills et al

. (1992)

Crassostrea gigas
Pacific

oyster Veliger LC50 Mortality Mortality 1 hr 556.0 Chien and Chow, 1989 Molluscs

Mercenaria

mercenaria
Hard Clam Larvae LC50 Mortality Mortality 48 hrs 5.0 Be llanca and Bailey,
1977

Asellus aquaticus Aquatic sowbug Mature,

non

reproducin

g LC50 Mortality Mortality 24 hrs 754.0 Kaniewska

Prus, 1982 Crustaceans

Callinectus

sapidus
Blue crab Adult LC50 Mortality Mortality 48 hrs 750.0 Vreenegoor
et al . 1977 Chlorine gas (Cl 2 )

Algae Chlorophyce

ae Green Algae 8 species NR Populatio

n Abundance 24 hrs

Static
system 100.0 Toetz et al

. 1977

Fish Clupea

pallasii
Pacific herring Juvenile LC50 Mortality Mortality 96 hrs 65.0 Thatcher,
1978

Lepomis
macroch irus Bluegill Juvenile LC50 Mortality Mortality 96 hrs 580.0 Office of pesticide

programs, 2000

Dreissena polymorpha
Zebra mussel Adult LT95 Mortality Mortality 17.7

hrs 90.0 Harrington

et al . 1997

Crassostrea virginica
American
oyster Larvae

(7 days) LC50 Mortality Mortality 0.5 hrs 120.0 Capuzzo,
1979

Clibanarius

humilis
Hermit crab NR LC50 Mortality Mortality 96 hrs 2,500.0 Best, 1991 Sodium hypochlor
ite

Molluscs

Hemigrapsus
nudus
Shore crab Adult LC50 Mortality Mortality 96 hrs 1,418.0 Thatcher,
1978MEPC 56/2/3

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- 3.doc Test

water

substance Organisms Scientfic
Name

Com mon Name Stage Toxicity Endpoint Effect Effect
Measurement Durati

on Exp. Concentrat

ion ( m g/L) Reference

Chlorella
sorokiniana Green algae 225

cells/ml NR Mortality Mortality 20 hrs 200.0 Kott and Edlis, 1969 Crusta ceans

Plankton Chaetocero s, Gracilis,

Dunaliella LC50 Mortality Mortality 96 hrs 90.0 Best, 1991

3.4.2 Endocrine disruption

No information is av

ailable on endocrine disruption by chlorine

3.4.3 Sediment toxicity

Chlorine may react with soil and sediment components to form chlorides, which, depending on their water solubility, are easily washed out. (Seiler et al . 1988 from US EPA database).

3.4 .4 Bioavailability/biomagnification/bioconcentration

There is no potential for the bioaccumulation or bioconcentration of chlorine (HSDB

, 1994).

3.4.5 Food web/population effects

As there is no potential for bioconcentration of chlorine, animals high in the food chain will not

be affected.

3.4.6 Data on mammalian toxicity

Sodium hypochlorite solution causes moderate mucosal irritation, the extent of which depends on the volume ingested, the viscosity and concentration of the preparation and the duratio n of the contact. When swallowed it may cause gastrointestinal irritation, with nausea, vomiting and diarrhea. Haematemesis may occur with concentrated solutions.

3.4.6.1 Acute toxicity

In humans, oral doses of 2.5 mg chlorine/day, administered to 10 men for 12 weeks, had no adverse effects (U.S. EPA 1990). No adverse effects were noted in persons ingesting water containing 50

- 90 ppm of chlorine (~1.4 to 2.6 mg Cl/kg/day) for a short period of time (U.S. EPA 1989). Drinking water concentrations of >90 ppm chlorine caused irritation of membranes of throat and mouth (U.S. EPA 1989). Concentrations of chlorine in the drinking water of greater than 25 ppm make the drinking water unpalatable (U.S. EPA

, 1989).

Chlorine is considered a primary irritant to th e mucous membranes of the eyes, nose, and throat and to the linings of the entire respiratory tract (Stokinger 1982). The extent of acute injury to humans depends on the concentration and duration of exposure as well as the water content of the

tissue inv olved and the presence of underlying cardiopulmonary disease (HSDB 1994). The estimated clinical effects of varying concentrations of chlorine are as follows: mild mucous

membrane irritation at 1

- 3 ppm; moderate irritation of the upper respiratory tract a t 5

- 15 ppm; immediate chest pain, vomiting, dyspnea, and cough at 30 ppm; toxic pneumonitis andMEPC 56/2/3

ANNEX Page 28

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- 3.doc pulmonary edema at 40

- 60 ppm; death at 430 ppm for 30 minutes or 1000 ppm for a few minutes (HSDB 1994). Seventy

- six individuals, exposed during a football gam e when approximately 1

, 100 pounds of chlorine gas were released from a plant, suffered no serious or prolonged incapacitation due to the release (HSDB 1994). If one survives acute exposure to chlorine, recovery is usually complete and rapid (U.S.EPA 1989)

.

In animals

- LC50 values for rats and mice are estimated to be 293 ppm for 1 h and 137 ppm

for 1 h, respectively (HSDB 1994). LCLo values for other species range from 330 ppm for 7 h (guinea pigs) to 660 ppm for 4 h (cats and rabbits) (U.S. EPA 1989). Mice and rats exposed to

chlorine at the RD 50 concentration (9

- 11 ppm, 6 h/day for 1, 3, or 5 days) developed degeneration of olfactory sensory cells in the olfactory mucosa, loss of cilia of the respiratory epithelium, and cellular exfoliation, primarily of the naso

- and maxilloturbinates (HSDB 1994). Signs and symptoms of the acute oral toxicity of chlorine in rats include decreased blood glutathione (30 minutes after 0.2 mg chlorine/kg administered as HOCl); decreased hypothalamic norepinephrine levels and increases in normetanephrine levels (3 and 24 hours after intubation

of 250 mg/kg free chlorine as HOCl); morphological and biochemical liver changes (within 2 days of dosing with 142.9 mg/kg free chlorine as NaOCl); kidney enlargement (200 mg/kg/day available chlorine for 14 days) (U.S. EPA 1989); and dose

- related increase in liver and kidney weights (210 mg dietary chlorine/kg body weight/day for 28 days) (U.S. EPA 1987). C57Bl/6N mice given chlorine (25

- 30 ppm) in the drinking water for one to thre e weeks had significant decreases in the number of peritoneal exudate macrophages, and decreased in vitro cytotoxicity against mouse melanoma (B16) and fibrosarcoma (UV

- 112) target cells (U.S. EPA 1989).

3.4.6.2 Effects on skin and eye

Free active chlori

ne can cause mild skin irritation to corrosive injury depending on the duration of the contact, concentration and pH of the solution. An 8% sodium hypochlorite solution has been shown to not cause skin irritation. Mild eye irritation to corrosive injury c an a lso be caused depending on the concentration and pH of the solution and the duration of contact.

3.4.6.3 Repeated

- dose toxicity

See chronic toxicity.

3.4.6.4 Chronic toxicity

The major target organs for the subchronic/chronic toxicity of chlorine in humans are the respiratory tract and the blood. The major target organs for the subchronic/chronic toxicity of chlorine in animals are the immune system, the blood, the cardiovascular system and the respiratory tract. The US EPA has derived an oral Rf D (reference dose) of 0.1 mg/kg/day for chlorine, based on a no

- observed

- adverse

- effect level of 14.4 mg/kg/day in a chronic drinking water study in rats.

Fifty

- five workers exposed to about 1.0 ppm of chlorine, 8 h/day for at least 90 days exhibited

asth ma, chronic bronchitis, tuberculosis, and emphysema (U.S. EPA 1987). The concentration

of 1 ppm is roughly equivalent to 0.415 mg/kg/day. In one case study, exposure to 0.015 mg/L of chlorine, 8 h/day for 6 years resulted in dyspnea, marked emphysema of both lower lung lobes, and reduced respiratory mobility (U.S. EPA 1987). Fifteen male workers exhibited

decreased residual levels of maximal mid

- expiratory flow rates and FEV/FVC after exposure

to 0.18 ppm chlorine, 8 h/day, 5 days/week for 8.9 years (U.S . EPA 1990). WorkersMEPC 56/2/3

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- 3.doc in 25 chlor

- alkali plants (332 males), exposed to chlorine concentrations of 0.006 to 1.42 ppm (8

- h TWAs) for about 10.9 years showed no dose

- related correlation between chlorine exposure and the prevalence of colds, dyspnea, palpitat ion, chest pain, or permanent lung damage (544 exposed workers had chest x

- rays) (Stokinger 1982 in US EPA, 1994).

Animals

- Groups of 70 F344/N male and female rats were given drinking water containing up

to 275 ppm of chlorine (14.4 mg/kg/day) for 2 y ears. No hematologic abnormalities were detected at interim sacrifices (14 and 66 weeks) and no non neoplastic abnormalities were observed in extensive gross and microscopic examinations at the end of the study (NTP 1992).

The EPA used the no

- observed

- ad verse

- effect level, 14.4 mg/kg/day, to calculate a chronic oral RfD of 0.1 mg/kg/day for chlorine (U.S. EPA 1994).

Sprague

- Dawley rats, given 5, 15, or 30 ppm chlorine in drinking water from weaning to

12 weeks of age exhibited significantly reduced splee n weights, suppressed delayed

- type hypersensitivity reactions, and decreased oxidative metabolism by macrophages at the highest dose; no effects were noted for the other doses or on other parameters of immunotoxicity (NTP 1992). F344 rats given doses of c hlorine up to 60 mg/kg/day for 92 days exhibited no histopathology in major organs (U.S. EPA 1989). Other species, exposed subchronically to chlorine exhibited the following: mice had no adverse effects on weight gain, food or water consumption, histolog

ical parameters (12.5 mg/kg/day for 50 days or 25 mg/kg/day for 33 days) (U.S. EPA 1989); New Zealand rabbits had increased hydroxyproline levels in heart tissue (1.6 mg chlorine/kg/day for 3 months; no effect at 0.1 mg/kg/day) (U.S. EPA 1989); Carneau pi geons exhibited cardiovascular effects (10 mg chlorine/kg/day in drinking water for 9 months) (U.S. EPA

, 1989).

In various studies, Sprague Dawley and F344 rats treated with chlorine orally for 12

- 24 months at doses ranging from 0.14 to 22 mg/kg/day exhib ited one or more of the following symptoms: significant decreases in red blood cell count and hematocrit (reversed at 6 months), increased osmotic fragility, increased mean corpuscular hemoglobin, significant decreases in body and liver weights, decreased brain and heart weights (p<0.05), and decreased salivary gland and kidney weights (U.S. EPA 1989), decreased spleen weight, and decreased thyroid weight (U.S. EPA

, 1987).

Rhesus monkeys (4/sex/dose) exposed to ó2.3 ppm of chlorine 6 h/day, 5 days/week fo r 1 year experienced ocular irritation, conjunctival irritation, hyperplasia of the nasal mucosa, and, in some females, epithelial hyperplasia of the trachea with minimal evidence of early nonkeratinizing squamous metaplasia (U.S. EPA

, 1990).

3.4.6.5 Deve lopmental and reproductive toxicity

No conclusion on the developmental/reproductive toxicity of chlorine can be made based on the

limited information available from human and animal studies.

Humans

–No information was found in the secondary sources sea rched to indicate that chlorine is a developmental/reproductive toxicant in humans.

Animals

–Female Sprague

- Dawley rats were given chlorine concentrations of 0, 1.0, 10,

or 100 mg/L in the drinking water (as HOCL) for 2.5 months prior to conception and t hroughout gestation, were killed on gestation day 20, and the fetuses were examined (NTP 1992 in

U.S. EPA

, 1994). Resorption was not increased at any concentration; however, fetuses inMEPC 56/2/3

ANNEX Page 30

I: \ MEPC \ 56 \ 2

- 3.doc the 100 mg/L group had soft tissue defects, including improper orienta tion of the heart and adrenal agenesis, and a slightly increased incidence of skeletal variants, such as incompletely ossified or missing sternebrae or rudimentary ribs. Statistical analyses were not available.

In other studies, chlorine administered to pregnant mice was negative for reproductive and teratogenic effects when given in drinking water as 10

- 13 ppm of sodium hypochlorite and hydrochloric acid (to maintain water pH of 2.5) or when given in heavily chlorinated municipal drinking water (U.S. EPA

, 1989).

Sperm head abnormalities were observed in B6C3F1 mice given 1.6 and 4.0 mg/kg/day of

chlorine as OCl

- , but not at 8.0 mg/kg or when given as HOCl (U.S. EPA 1994). BDII rats (236 total males and females) given 10 mg/kg/day of free residual chlor ine in the drinking water throughout 7 generations had no adverse effects on weight gain, food consumption, water consumption, fertility, lifespan, growth pattern, hematology, histology (liver, spleen, kidney or other organs) (U.S. EPA

, 1989).

3.4.6.6 Mut agenicity/genotoxicity

Chlorine was mutagenic in Salmonella typhimurium strains TA1530 and TA100, without metabolic activation; produced chromosome aberrations in human lymphocytes and other mammalian cells (ò20 ppm); interacted with DNA in E. coli polA test (as sodium hypochlorite); and was negative for the induction of erythrocyte micronuclei or chromosome aberrations of

bone marrow cells of Swiss CD

- 1 mice (up to 8 mg/kg/day of NaOCl) for up to 5 days (U.S. EPA

, 1989).

3.4.6.7 Toxicokinetics

No addi tional information is available on toxicokinetics for chlorine.

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

3.4.7.1 Modes of degradation (biotic; abiotic)

Chlorine hydrolyzes very rapidly in water (rate constants r ange from 1.5 x 10

- 4 at 0øC

to 4.0 x 10

- 4 at 25øC; half

- life in natural waters, 0.005 s) (U.S. EPA 1989). In fresh and wastewaters at pH >6, complete hydrolysis occurs with the formation of hypochlorous acid (HOCl) and chloride ion (Cl

- ). The hypochlorou

s acid ionizes to hydrogen ion (H+) and hypochlorite ion (OCl

- ). At pH values >5, OCl

- predominates; at pH values <5, HOCl predominates (U.S. EPA 1989). Free chlorine (Cl2, HOCl, and OCl

- ) reacts rapidly with inorganics such as bromide and more slowly wi th organic material present in natural waters. These reactions yield chloride, oxidized organics, chloroganics (including trihalomethanes), oxygen, nitrogen, chlorate, bromate and bromoorganics.

3 .4.7.2 Bioaccumulation, octanol/water partition coefficien t

There is no potential for the bioaccumulation or bioconcentration of chlorine (HSDB 1994).

3.4.7.3 Reaction with organic matter

Chlorine reacts slowly with organic material present in natural waters. These reactions yield chloride, oxidized organics, chloroganics (including trihalomethanes), oxygen, nitrogen, chlorate, bromate and bromoorganics.MEPC 56/2/3

ANNEX Page 31

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- 3.doc 3.4.7.4 Potential physical effects on wildlife and benthic habitats

Chlorine has been shown to have high acute toxicity to many organisms (U.S. EPA, 1994). Low level chlorination (0.05 to 0.15 mg/L) results in significant shifts in the species composition of marine phytoplankton communities (HSDB 1994 in U.S. EPA, 1994). Chlorine is phytotoxic but is also essential to plant growth; crops need around 5 pound s or more of chlorine per acre.

Acute toxicity to plants is characterized by defoliation with no leaf symptoms and, in higher plant forms, by spotting of the leaves (at 1.5 mg/m 3 ) and marginal and interveinal injury (at 150

- 300 mg/m 3 ) (Seiler et al . 1988 in U.S. EPA, 1994). No data were found regarding the toxicity of chlorine to terrestrial animals; however, the data from experimental studies indicate that injury to animals would occur only in the presence of high concentrations of chlorine, either

in dr inking water or the ambient atmosphere (U.S. EPA, 1994).

As chlorine and chlorine compounds do not persist in sediments it is not expected to affect benthic habitats unless directly applied in high concentrations.

Potential residues in seafood

As there i s no potential for the bioaccumulation or bioconcentration of chlorine there is no risk of potential residues in seafood.

3.5 Hydrogen (Related Chemical)

Note that hydrogen gas is produced in the electrochemical cell of the RBT reactor as a byproduct

of hydroxyl radicals. B

asic A pproval has been recommended by the GESAMP

- Ballast Water Working Group (GESAMP

- BWWG) (MEPC 55/2, 2006) for use of similar electrochemical systems designed for the treatment of ballast water that also produce hydrogen gas

. Full d etails on the nature, toxicity and risks posed by this compound is nonetheless provided in this report for completeness.

3.5.1 Acute aquatic toxicity

There is no data on acute aquatic toxicity of hydrogen.

3.5.2 Endocrine disruption

None of the availab le data indicate toxicity for exposures of any duration.

3.5.3 Sediment toxicity

There is no data on sediemnt toxicity of hydrogen.MEPC 56/2/3

ANNEX Page 32

I: \ MEPC \ 56 \ 2

- 3.doc 3.5.4 Bioavailability/biomagnification/bioconcentration

Hydrogen does not bioconcentrate in the tissues of organisms.

3.5.5 Food web/population effects

Hydrogen does not bioconcentrate or accumulate in the tissues of organisms therefore there is no risk to organisms up the food chain.

3.5.6 Data on mammalian toxicity

None of the available data indicate toxicity for ex posures of any duration. Hydrogen is not listed by NTP, OSHA or IARC.

3.5.6.1 Acute toxicity

High concentrations of hydrogen in the air can cause a deficiency of oxygen with the risk of unconsciousness or death. Ingestion is not an observed route of ex posure to gaseous hazardous materials. Exposure to moderate concentrations may cause dizziness, headache, nausea and

unconsciousness.

3.5.6.2 Effects on skin and eye

No detrimental effects of skin or eye contact has been reported. Hydrogen only poses a risk of thermal burns if ignited.

3.5.6.3 Repeated

- dose toxicity

None of the available data indicate toxicity for exposures of any duration.

3.5.6.4 Chronic toxicity

None of the available data indicate toxicity for exposures of any duration.

3.5.6.5 Developmental and reproductive toxicity

None of the available data indicate toxicity for exposures of any duration.

3.5.6.6 Mutagenicity/genotoxicity

None of the available data indicate toxicity for exposures of any duration.

3.5.6.7 Toxicokinetics

N one of the available data indicate toxicity for exposures of any duration.MEPC 56/2/3

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- 3.doc 3.5.7 Data on environmental fate and effect under aerobic and anaerobic conditions

3.5.7.1 Modes of degradation (biotic; abiotic)

Hydrogen exists naturally in the atmosphere and will be dissipated rapidly in well ventilated

areas.

3.5.7.2 Bioaccumulation, octanol/water partition coefficient

Hydrogen rapidly dissipates so there is no bioaccumulation.

3.5.7.3 Reaction with organic matter

Hydrogen is not known to react with orga nic matter.

3.5.7.4 Potential physical effects on wildlife and benthic habitats

No adverse ecological effects are expected. Hydrogen does not contain any Class I or Class II

Ozone depleting chemicals (40 CFR Part 82). Hydrogen is not listed as a marine pollutant by DOT (49 CFR Part 171).

3.5.7.5 Potential residues in seafood

Hydrogen does not bioaccumulate so there is no risk of hydrogen residues in seafood.

4 PHYSICAL AND CHEMICAL PROPERTIES FOR THE ACTIVE SUBSTANCES
AND PREPARATIONS AND TREATED BAL LAST WATER

Data on the physical and chemical properties of the Active Substance s, preparations and treated ballast water are provided in Table 2.

5 USE OF THE ACTIVE SUBSTANCE OR THE PREPARATION

5.1 Active Substance P reparation

The RBT reactor employs the use of two Active Substance s in its working principals. These substances namely; Ozone and sodium hypochlorite, are generated on board the vessel from natural sources such as air from the atmosphere for Ozone production, and sodium chloride or

other e lectrolyte substance found in the ballast water being pumped.

The main reason for the addition of these substances is to assist with the cavitation that forms the basis of the reactor; also these chemicals have the additional advantage of maintaining a st erile environment within the ballast tanks on board a ship.

Both Ozone and sodium Hypochlorite have been used to disinfect drinking water for many decades. The use of these chemical compounds for potable water treatment is well documented.

These chemical

substances are added into the ballast water during ballasting operations and as such do not need to be handled by the crew at any stage.MEPC 56/2/3

ANNEX Page 34

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- 3.doc Levels of these chemical are kept to a minimum i.e. 0.001mg/l for the O3 gas and <1.0ppm for the sodium Hypochlorite

.

No additional preparation methods or handling of these chemicals is required. The RBT Reactor is supplied pre

- calibrated for the specific ballast flow rate as determined by the individual vessel requirements.

5.2 The RBT “Coronatech” Ozone generator

Ozone gas that is produced and subsequently infused into the ballast water treatment system is generated by the corona method.

The corona discharge generator utilizes a high frequency transformer (typically 16

- 20KHz) This method is favoured because greate r Ozone production can be achieved with less electrode surface area and less electrical consumption (0.75 amps)

.

The RBT Ozone discharge generator accelerates electrons so as to give them sufficient energy to split the oxygen

- oxygen double bond upon impac t with another oxygen molecule. The two oxygen atoms, which are produced by the collision, react with other diatomic oxygen molecules

to form Ozone

. Ozone is produced as a result of an electrical discharge.

Various factors that affect the production of Oz one gas via the generator have been taken into account i.e., temperature, humidity and oxygen supply. In order to maintain the efficient working temperature for the effective production of Ozone gas, a cooling system is integrated into the equipment. Filte red atmospheric air is cooled via a chiller; the chiller also acts to cool the corona discharge chamber. Ozone produced via the generator is approximately 2%.

5.2.1 R isks to ship safety and crew

The Ozone generator that forms part of the RBT ballast wate r treatment system is purpose built, baring in mind the safety of the ship its crew and the environment.

Specific consideration was given to the assurance that no Ozone leaks could or would take place during the normal operation of the O3 generator.

Safe ty features

Ozone is produced from the surrounding air, and is drawn into the ballast water by means of a venturi system. (Vacuum) As such the O3 generator will not produce any additional O3 gas in

the event of a zero flow of air.

Two one

- way / non

- retur n, check valves are installed. One check valve on the Ozone delivery

line and one check valve on the air intake of the Ozone generator. The purpose of these check valves is to ensure a one directional flow of air through the O3 generator and also that no

O zone gas can be dissipated into the surrounding air. In the most unlikely event that the

check valves should fail, ballast water would be passed back into the corona tubes, thereby rendering the Ozone generator incapable of producing any O3 gas. An 5 mill i

- amp fuse protects

the O3 generator.MEPC 56/2/3

ANNEX Page 35

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- 3.doc Corrosion

Due to the fact that the RBT reactor makes use of only 0.0001 ppm of Ozone gas in its working principals no noticeable increase in corrosion of the ballast pipe work or tanks will occur. Surface coatings w

ithin the ballast tanks offer additional protection from any residual Ozone gas.

Venting

In the unlikely event of any Ozone gas build up within the ballast tank, any free Ozone gas that is

not absorbed into the ballast water will be vented via the balla st tank venting system. At no time

will the free Ozone reach any harmful or dangerous levels.

System f ailure

The Ozone generator has no moving parts and as such is maintenance free. In the event of Ozone system failure, warning signals are transmitted to the bridge and ballast control room.

5.3 Electro Chemical Production

Installed into the electrical control panel is an electrical rectifier that supplies a current to the electrodes that form part of the RBT reactor. Passing an electrical potential thr ough a cathode and an anode produces Electro chemical production of chemical compounds such as Sodium Hypochlorite and gases. The dissolved salts and compounds within ballast water, namely Na Cl serve as an electrolyte in the process, produce compounds suc h as Hydrogen, Oxygen and Sodium Hypochlorite. The electrodes that form part of the RBT Reactor are selected from the group VIII metals on the periodic table

–Ruthenium, Titanium.

Low concentrations of Sodium Hypochlorite are produced at the surface of t he electrodes (< 1 ppm) Residual chlorine content assists in maintaining a sterile environment within the ballast tank. Other useful chemicals produced by the electrodes that are harmful or lethal to marine organisms include, Bromine, Hypobromous acid, Chl orine Dioxide, Chlorine, Hypobromite, Hydroxyl radical. These chemicals degrade over a period of time and would not be harmful to the environment when the ship de

- ballasts.

Chlorine residuals are affected by temperature and pH of the water being treated, the effectiveness of chlorine decreases with an increase of pH above 8.0. For human consumption chlorine residuals of 1.2 to 1.5 ppm are acceptable. It is important to maintain chlorine (NaOCl) limits within the prescribed limits for human consumption, as no particular limits and standards are currently available for the treatment of seawater. The levels of NaOCl produced by the RBT reactor are less than 1 ppm.

Sodium Hypochlorite levels fall within the specifications of the paint manufacturers responsible for the manufacture and application of ballast tank paint protection systems

5.3.1 Risks to ship safety and crew

The rectifiers installed into the electrical control panel of the RBT reactor supplies a DC current directly to the electrodes of the reacto r.MEPC 56/2/3

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- 3.doc As with all electrical component that make up the treatment process of the equipment, ship and crew safety are taken into account. Where necessary the electrical equipment installed would be supplied with the necessary certification and build to ATEX S tandards, should the equipment

need to be installed into a hazardous area on board a vessel.

Safety features

Certain chemicals predominantly “sodium hypochlorite,” are produced inside of the RBT reactor directly from the seawater, when the ballast water is being pumped into the vessel. The ship crew requires no additional handling of any chemicals.

Circuit breakers and fuses are installed into the ECP that would trip in the event of any direct

short circuit of the rectifiers.

Corrosion

Chemicals produc ed by the electro chemical components of the RBT reactor can be potentially corrosive, however, levels of these chemicals and gases are low and will not increase the corrosion rate of the ballast piping and tanks.

Venting

Any un

- dissolved gases such as c hlorine gas and hydrogen will be vented through the ships ballast tank vents. Hydrogen gas levels would never reach dangerous levels (4%)

.

System failure

In the unlikely event of a rectifier or electrode failure, aural and visual alarm warnings are

trans mitted to the ballast control room as well as the bridge. Two rectifiers are installed to ensure that ballasting operations would not be affected in the event of a rectifier failure.

6 RISK ASSESSMENT

6.1 Risk to safety of ship

There is an increased ris k of corrosion arising form the oxidizing properties of Ozone . However,
because Ozone has such a short half

- life (< 6 seconds in seawater), it will only exist for a very

short time in the ballast tank.

Mitigation required will be to ensure that the mach inery producing the Ozone and the piping leading into the ballast water is coated with a ceramic coating and/or constructed of anti

corrosive material such as stainless steel. Ozone is the most corrosive of the Active Substance s applied directly to the ba llast water and therefore any measures taken to prevent corrosion by

Ozone will be sufficient to prevent corrosion by the other Active Substance s. Hydrogen poses a flammability risk but quantities produced will be so small as to be of no concern. Hydroxy l radicals degradate within nanoseconds and are therefore of no concern with regards to corrosion.

There are no risks associated with the handling of active chemical as the Active Substance s and the relevant chemicals are produced in situ and are therefor e not handled.MEPC 56/2/3

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- 3.doc Ozone Ship Risk Summary

Risk of substance: Medium Certainty: High

Risk after mitigation: Very Low Certainty: High

Bromate ion Ship Risk Summary

Risk of substance: Low Certainty: High

Risk after mitigation: Very Low Certainty: High

Bro moform Ship Risk Summary

Risk of substance: Low Certainty: High

Risk after mitigation: Very Low Certainty: High

Sodium hypochlorite (free acti ve chlorine) Ship Risk Summary

Risk of substance: Medium Certainty: High

Risk after mitigation: Very Low

Cert ainty: High

Hydrogen Ship Risk Summary

Risk of substance: Medium Certainty: High

Risk after mitigation: None Certainty: High

Hyd roxyl radical Ship Risk Summary

Risk of substance: None Certainty: High

Risk after mitigation: None Certainty: HighMEPC 56/2/3

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- 3.doc 6.2 Ri sks to human health

6.2.1 Ozone

Ozone is very toxic by inhalation route and there is also a potential for Ozone to cause mutagenicity after oral exposure. Therefore strict mitigation measures will have to be applied to avoid human exposure.

Mitigation: The Ozone generator is fabricated in such a way as to ensure that no Ozone gas can escape into the surrounding air. When the ballast pump is incited the Ozone generator is turned on automatically, and the Ozone gas that is generated is sucked directly int o the ballast water being treated.

A non

- return /one way valve, situated onto the corona reactor ensures that no Ozone gas escapes. Should there be no air flow through the corona reactor, no Ozone gas is produced thereby ensuring that the equipment is saf e for the ship and its crew. Even in the event of accidental or purposely turning on of the Ozone generator it would be impossible for the Ozone generator to produce Ozone gas without an airflow through the corona reactor. The air flow required is

created by a venturi that ensures that all of the Ozone gas is infused into the ballast water.

In addition, with regard to the health of the ship crew, placards relating to the general hazards of

Ozone inhalation should be placed onboard to promote general aware ness.

Ozone Human Health Risk Summary

Risk of substance: Low Certainty: High

Risk after mitigation: Very Low Certainty: High

6.2.2 Bromate ions

Several studies have indicated high potential for mutagenicity and carcinogenicity in humans.

The current M

CL (Maximum Contaminant Level) in the United States is 0.010mg/L and the MCL goal is zero because of the possibility that bromate may function as a genotoxic carcinogen. Therefore human exposure should be totally avoided.

Mitigation: Bromate ions will for m in the ballast water which is pumped into the ship’s ballast tanks mechanically and therefore there will be no human contact on board the ship with treated ballast water. Providing the treated water is released more than 12 hours (the half life of a

brom ate ion) after it was treated, bromate ions will pose no threat to human health outside of the ship either.

Bromat e ion Human Health Risk Summary

Risk of substance: Low Certainty: High

Risk after mitigation: Very Low Certainty: HighMEPC 56/2/3

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- 3.doc 6.2.3 Bromoform

Br omoform is a highly reactive compound capable of bioacumulating. Exposure to bromoform vapors may cause irritation to the respiratory tract, lacrimation, and liver damage. Bromoform has also been classified as a Group B2, probable human carcinogen (US EPA, 2000). Exposure

should therefore be minimized.

Mitigation: According to tests conducted on water that was treated using Resource Ballast Technologies (Pty) Ltd., the concentration of all trihalomethanes (which was made up almost entirely of bromoform) wa s 19.6 μ g/L. The acceptable concentration level of total trihalomethanes in the United States for water used by humans is 80.0 μ g/L. Therefore the concentration levels of bromoform produced by this methodology are unlikely to pose a health

risk to humans. Ho wever, further testing is recommended with seawater from a variety

of sources.

Brom oform Human Health Risk Summary

Risk of substance: Low Certainty: High

Risk after mitigation: Low Certainty: High

6.2.4 Free a ctive c hlorine

Swallowing sodium hypochlor ite can cause irritation, pain and inflammation of mucoses. In severe cases it can cause ulceration and perforation of the gastro intestinal tract. Vapours are not easily formed although under certain conditions, high concentration solutions in high pH mix ed with acids, may produce chlorine gas which causes mild irritation of the nose and throat.

Mitigation: The sodium hypochlorite will be produced in situ and will be pumped directly into the ballast water. Therefore there will be no potential risk of the crew being exposed to chlorine in any form. Chlorine has a rapid half life (0.005) and does not bioaccumulate. Therefore it poses

no risk to humans once after the treated ballast water has been released.

Free active chlorine human health risk summary

Ri sk of substance: Medium Certainty: High

Risk after mitigation: None Certainty: HighMEPC 56/2/3

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- 3.doc 6.2.5 Hydrogen

Hydrogen is not toxic but if released in large enough quantities it may displace oxygen leading to unconsciousness or asphyxiation. It is also highly exp losive in high concentrations.

Mitigation: The amount of hydrogen produced by the electro

- chemical electrodes is miniscule and would not reach any dangerous levels, any exce ss that is not already absorbed /dissolved into

the ballast water would be vented i nto the atmosphere via the vents in the ballast tanks.

Hydrogen human health risk summary

Risk of substance: Medium Certainty: High

Risk after mitigation: Very Low Certainty: High

6.2.6 Hydroxyl radicals

Hydroxyl radicals have an extremely raid half life (nanoseconds) and do not persist or bioaccumulate. They therefore do not pose a risk to human health.

7 RISKS TO THE AQUATIC ENVIRONMENT

7.1 Screening for persistence, bioaccumulation and toxicity

Active Substance s and relevant chemicals produced o r introduced by the RBT reactor for treatment of ballast water include the following:

• Ozone

,

• bromate ions,

• bromoform,

• hydroxyl radicals,

• chlorine gas (Cl 2 ),

• free active chlorine species (HOCl and ClO

- ), and

• hydrogen gas

.

Of these, Ozone , hydroxyl radical s and chlorine compounds (chlorine gas and free active chlorine species) are extremely short lived and have very little or no potential for persistence or for bioaccumulation (see data presented in the preceding sections). Thus, although their toxicity

to marine life may be high (which is the reason for their use in the RBT reactor) risks posed to the environment through the release of treated ballast water is considered negligible.

Bromate ions are only likely to be formed in seawater but have a longer h alf life than Ozone

, hydroxyl radicals and the chlorine compounds mentioned above (approximately 12 hours).

The octanol/water particial coefficient log value ( P ow ) for bromate is

- 7.18 and such it is not considered to pose a risk of concentration in marin e organisms or to animals higher up the

food chain (PAN Pesticides Database). Acute toxicity of bromate is also low (approximately 1/1000 of that of Ozone ) and concentrations in ballast water treated with the RBT reactor are very low relative to concentra tions at which toxic effects have been demonstrated (see data presented in the preceding sections).MEPC 56/2/3

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- 3.doc Bromoform has an octanol/water partition coefficient (P
ow ) of approximately 2.38 which suggests that it will tend to partition to fat tissues of aquatic or ganisms, and hence may be slightly bioconcentrated by aquatic organ isms. The concentration of bromoform in ballast water treated with the RBT reactor is very low (<20 μg/l) relative to concentrations at which toxic effects have been detected (9 000

- 140 000 μg/l, see preceding sections).

In conclusion, therefore it is cl ear that none of the Active Substance s or relevant chemicals produced by the RBT reactor can be considered as PBT substances ( Table 7 ).

Table 7

. Risk characterization for Active Substance s and relevant chemicals

produced or introduced by the RBT reactor

Substance Persistence (Half

- life: > 60 days in marine water/> 40 days in freshwater, or > 180 days in marine sediment, or > 120 days in freshwater sediment Bioaccumulation (Log Pow >3) Toxi city (Chronic
NOEC < 0.01 mg/l)

Ozone No No Yes

Bromate ions No No Yes Bromoform, Yes No Yes Hydroxyl radicals No No Yes Chlorine gas (Cl 2

) No No Yes

Free active chlorine species (HOCl and ClO

- ) No No Yes

Hydrogen gas No No No

7.2 Evaluation of t he treated ballast water

Toxicity test has not yet been undertaken on samples of treated ballast water due to time constraints but it is the intention of the proponents to undertake relevant toxicity tests as required by the GESAMP

- Ballast Water Working G roup (GESAMP

- BWWG) (MEPC 55/2, 2006).

7.3 Aquatic toxicity of Active Substance & Relevant Chemical

7.3.1 Acute aquatic toxicity

An overview of acute toxicity data (lowest short

- term LC 50 test values) for up to three taxa (fish, crustacean, mollusc, alg ae) where available are presented in Table 8.

It can be concluded from these data that acute toxicity to Ozone can be seen at >500 μg/L, bromate ions at >176 000 μg/L, bromoform at >16 000 μg/L, chlorine gas at >600 μg/L, and sodium hypochlorite at around 100 μg/L. The concentration of bromoform and chlorine in ballast water treated with the RBT reactor are low (<20 μg/l, and 80

- 320 μg/l, resepctively) relative to levels at which acute toxicity can be expected.MEPC 56/2/3

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- 3.doc Table 8

. Overview of acute toxicity data (lowe st short

- term LC 50 test values)

Test
Substance F

C

A Species Effect Value (μg/L)

F L epomis macrochirus 24h LC 50 60

M Dreissena polymorpha 5.8h LC 50 520 Ozone

A

- - -

F Oncorhynchus keta 96h LC 50 512 000

C Neomysis awatschnsis 24h LC 50 176 000 Bromate

I on

A

- - -

F Cyprinodon variegates 48h LC 50 16 000

C Peneus aztecus 96h LC 50 20 000 Bromoform

A

- - -

F Morone saxatilis 6h LC 50 610

C Asellus aquaticus 24h LC 50 754 Chlorine gas (Cl 2 )

A

- - -

F Clupea pallasii 96h LC 50 65 Sodium hypochlorite M Crassotrea virginica 0.5 LC 50 120

F = Fish, C = Crustace

a, A = Algae, M = Molluscs

7.3.2 Chronic aquatic toxicity

An overview of chronic toxicity data (lowest long

- term LC 50 test values) for up to three taxa (fish, crustacean, mollusc, algae) where available are presented in Table 9 . It can be concluded from these data that chronic toxicity to Ozone and free active chlorine can both be expected

at >5 μg/L. Considering the short half lives of these substances, they are not expected to be

released into the environment above these levels.

Table 9

. Overview of acute toxicity data (low est long

- term NOEC test values)

Test
Subst ance F

C

A Species Effect Value (μg/L)

F Oncorhynchus mykiss 90d NOEC 5

C

- - - Ozone

A

- - -

F

- - - C

- - - Bromate

I on

A Skeletonema costatum 7d NOEC 0.125

- F Menidia penensulae eggs 28d NOEC 40

C Crassostrea v irginica & Rangia cuneata 15d NOEC 7 Free active
chlorine

A Phytoplankton 21d effect 1 F = Fish, C = Crustacea, A = Algae, M = MolluscsMEPC 56/2/3

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- 3.doc 7.3.3 Derivation of the Predicted No Effect Concentration (PNEC)

According to the GESAMP

- Ballast Water Working Group ( GESAMP

- BWWG) (MEPC 55/2,

2006 ), PNEC for Active Substance s and relevant chemicals should be derived by dividing

the lowest available effect concentration with an appropriate assessment factor, as provided in

Table 10

.

Table 10

. Effect assessment for deriving PNECs for fres hwater and saltwater

(MEPC 55/2)

Assessment Factor

Data set PNEC long PNEC

short

Freshwater assessment

Lowest short

- term L(E)C 50 from fresh water species representi ng three trophic levels 1000 10

- 100

Lowest chronic NOEC from three freshwater or saltwater species representing three trophic levels 10

Saltwater assessment

Lowest short

- term L(E)C 50 from marine species representing three trophic levels 1000 10

- 100

Lowest chronic NOEC from three freshwater or saltwater species representing three trophic levels 100

Lowest chronic NOEC from three saltwater species representing three trophic level 100

Lowest chronic NOEC from three freshwater or saltwater species representing three trophic levels + at least two

chronic NOECs from additional marine taxonomic

groups 10

Table 11

. Predicted concentrations of Active Substance and relevant chemical in

treated ballast water produced or introduc ed by the RBT reactor

Active
Substance /

chemical P

N

E
C Concentration immediately

after treatment (μg/L) Concentr

ation

after 2 h (μg/L) Concentr

ation

after 4h (μg/L) Concent

ration

after 8h (μg/L) Concentra

tion after

12h (μg/L)

Ozone (freshwater) 1 000 nd n d nd nd

Ozone (seawater) 1 000 62.5 3.9 0.015 5.9 x 10
- 5

Bromoform (seawater) 19.6 19.6 19.6 19.6 19.6

Chlorine gas (freshwater) 0.32 nd nd nd nd

nd = not detectable

.MEPC 56/2/3

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- 3.doc It is only relevant to evaluate PNEC values for Ozone in freshwate r and bromoform in seawater for this study (the remaining Active Substance s are either created in negligible concentrations in the other medium or decay to rapidly to be of consequence).

For Ozone there is only one chronic value available. An assessment factor of 100 is applied to this value resulting in a PNECchronic = 0.05 μg/L. There are 2 values available for acute data,

the lowest of which is 60 μg/L. Apply a factor of 1000 to this yields a PNECacute = 0.06 μg/L.

Of these two data the lowest value is taken as the resulting PNEC: PNEC = 0.05 μg/L as Ozone

. In the case of Ozone introduced to freshwater a total time of at least 8 hours is required before the ballast water can safely be discharged.

For bromoform there is one chronic value available. An assessment factor of 100 is applied to this value resulting in a PNECchronic = 48 μg/L and 3 for the acute data, the lowest of which

is 7.1 mg/L. Using an assessment factor of 1000 the PNECacute is established at 7.1 μg/L. Of

these two data the lowest v alue is taken as the resulting PNEC: PNEC = 7.1 μg/L as bromoform.

The half life of bromoform is in the order of 1

- 2 month and delaying release of ballast water is not considered feasible. Concentration in the treated ballast water (19.6 μg/L) are only

m arginally higher than the PNEC value (7.1 μg/L) and hence not considred to pose a major risk

to the environment.

8 LITERATURE CITED IN THIS APPLICATION

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Arizona Department of Health Services, Office of Environmental Health, Environmental Health Consultation Services. 2005. Prescott Water Company Evaluation of volatile organic compounds in residential area. Prescott Valley, Yavapai County, Arizona.

Asbury, C., and R. Coler. 1980. Toxicity of Dissolved Ozone to Fish Eggs and Larvae. J.Water

Pollution Control Fed. 52(7):1990

- 1996.

ATSDR (Agency for Toxic Substan ces and Disease Registry). 1990. Toxicologica l Profile for Bromoform and Chlorodibromomethane. Prepared by Life Systems, Inc., under Subcontract to Clement Associates, Inc., Contract No. 205

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Bellanca, M.A., and D.S. Bailey. 1977. Effects of Chlorinated Effluents on Aquatic Ecosystem in

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Buccafusco, R.J., S.J. Ells, and G.A. LeBlanc. 1981. Acute Toxicity of Priority Pollutants to Bluegill (Lepomis macrochirus). Bull.Environ.Contam.Toxicol. 26(4):446

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to Marine Animals

- A Preliminary Review. Mar.Pollut.Bull. 10(2):45

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- 84.

Crecelius, E.A. 1979 . Measurements of Oxidants in Ozonized Seawater and Some Biological Reactions. J.Fish.Res.Board Can. 36(8):1006

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Dandy, J.W.T. 1972. Activity Response to Chlorine in the Brook Trout, Salvelinus fontinalis (Mitchill). Can.J.Zool. 50(4):405

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- 1297). Hall, L.W.Jr., D.T. Burton, and L.B. Richardson. 1981. Comparison of Ozone and Chlorine Toxicity to the Developmental Stages of Striped Bass, Morone saxatilis. Can.J.Fish.Aquat.Sci. 38(7):752

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Harrington, D.K., J.E. Van Benschoten, J.N. Jensen, D.P. Lewis, and E.F. Neuhauser. 1997. Combined use of Heat and Oxidants for Controlling Adult Zebra Mussels. Water Res. 31(11):2783

- 2791.

Heitmuller, P.T., T.A. Hollister, and P.R. Parrish. 1981. Acute T oxicity of 54 Industrial Chemicals to Sheepshead Minnows (Cyprinodon variegatus). Bull.Environ.Contam.Toxicol. 27(5):596

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Paller, M.H., and R.C. Heidinger, 1979. The Toxicity of Ozone to the Bluegill. J.Environ.Sci.Health A14(3):169

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Perrins, J.C., Cooper, W.J., van Leeuwen, J.H., Herwig, R.P. 2006. Ozonation of seawater from different locations: Formation and decay of total residual oxi dant

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Toetz, D., L. Varga, and M. Pierce. 1977. Effects of Chlorine and Chloramine on Uptake of Inorganic Nitrogen by Phytoplankton. Water Res. 11(3):253

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– 1487.

von Gunten, U., Oliveras, Y., 1997. Kinetics of the reaction between hydrogen peroxide and hypobromous acid: Implication on water treatment and natural systems. Water Research 31, 900

– 906.

Vreenegoor, S.M., R.M. Block, J.C. Rhoderick, and S.R. Gullans. 1977. The Effects of Chlorination on the Osmoregulatory Ability of the Blue Crab, Callinectes sapidus. Assoc.Southeast.Biol.Bull. 24:93 (ABS No.13).

* * *MEPC 56/2/3

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- 3.doc APPENDIX 2MEPC 56/2/3

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- 3.doc _______________