Toxic Dinoflagellate Spores in Ships' Ballast Water

Final Report

FIRDC Grant 89 I 39

Toxic dinoflagellate spores in ships' ballast water :

A danger to aquaculture

G.M. Hallegraeff

CSIRO Marine Laboratories,

GPO Box 1538, Hobart, Tasmania 7 0 01

April 1992

Foreword

The present invꢁstꢃgations on "Toxꢃc dinoflagꢁllatꢁ sporꢁs in shꢃps' ballast wateꢇ" and "its implꢃcatꢃons foꢇ aquaculturꢁ" wꢁrꢁ fundꢁd by FIRDC gꢇant 89 I 39 (Sꢁpt 1989 - Sꢁpt 1 991)

.

Thꢃs ꢇꢁsꢁaꢇch involved a collaboꢇative effort bꢁtwꢁꢁn CSIRO Dꢃvꢃsꢃon of Fꢃshꢁrꢃꢁs and thꢁ Australian Quaꢇantꢃne and Inspꢁctꢃon ꢀꢁꢂꢃꢄꢁ (AQIS), and was instꢃgatꢁd by thꢁ ꢄlaim by CSIRO that thꢁ toxꢃc dꢃnoflagꢁllate Gymnodinium catenatum in Tasmanian waters could havꢁ bꢁꢁn introducꢁd vꢃa cyst stagꢁs ꢄontaꢃnꢁd ꢃn ships' ballast watꢁr. In Fꢁbruary 1986, contamꢃnatꢃon of Tasmanꢃan shꢁllfish wꢃth dinoflagellatꢁ toxꢃns lꢁd to the closurꢁ of 15 shꢁllfꢃsh farms for pꢁrꢃods up to 6 months. Subsꢁquꢁntly, sꢃmꢃlar toxic dꢃnoflagꢁllatꢁ outbrꢁaks suꢇfacꢁd in the Australꢃan ports of Adꢁlaꢃdꢁ (Aiexandrium minutum ) and

Mꢁlbourne (Aiexaꢁdrium catꢀꢁꢀꢂa )

.

Gꢁnꢁtꢃc ꢁvꢃdꢁnꢄꢁ (rRNA fꢃngꢁrprꢃnts) suggꢁst that thꢁsꢁ latter spꢁcꢃes arꢁ also ballast water ꢃntroduꢄtꢃons.

The presꢁnt ꢇꢁsꢁarch rꢁꢄꢁꢃvꢁd consꢃdꢁrablꢁ natꢃonal and
ꢃnteꢇnatꢃonal publꢃꢄꢃty ( front pagꢁ nꢁws in thꢁ Hobart "Mꢁrꢄury" and "Sydney Moꢇning Hꢁrald", natꢃonal tꢁlꢁvꢃsꢃon covꢁragꢁ on thꢁ "7.30 ꢇꢁport" and "Beyond.2000"). The Australꢃan Quarantꢃnꢁ and Inspꢁꢄtꢃon Sꢁꢂꢃcꢁ has respondꢁd to thꢃs ꢁvꢃdꢁnce by ꢃntroducing, as of 1 Fꢁbruary 1990, voluntary ballast water guidꢁlinꢁs for shꢃps entꢁring Auꢆtralian poꢅꢆ fꢇom ovꢁrsꢁas. As of 1 Novembeꢇ 1991, thꢁ Intꢁrnatꢃonal Marꢃtꢃmꢁ Organꢃsatꢃon (IMO) ratifꢃꢁd thꢁse guidꢁlꢃnes for adoptꢃon on an ꢃntꢁrnatꢃonal basꢃs.

The present FIRDC- fundꢁd rꢁsꢁarch has funꢄtꢃonꢁd as a ꢄatalyst for fuꢅheꢇ ballast watꢁꢇ ꢇꢁsꢁarch funds (600 K) made avaꢃlable by AQIS and BRR. Regulaꢇ meꢁtings (Nov. 1989, Feb.1990, June 1991, Oct. .1991, Nov. 1991, Feb. 1992) of a nꢁwly established Scientꢃfꢃc Workꢃng Group on Ballast Wateꢇ (undꢁꢇ diꢇꢁctꢃon of Dꢇ.Mꢁryl Wꢃlliams, BRR) arꢁ ovꢁrsꢁꢁꢃng thꢁ implementatꢃon of somꢁ of thꢁ ꢄonꢄlusꢃons of thꢁ prꢁsꢁnt rꢁsꢁarꢄh.

Oꢇ Gustaaf M. Hallꢁgraꢁff, CSIRO Dꢃvꢃsꢃon of Fꢃshꢁrꢃꢁs, GPO Box 1538, Hobart, Tas. 7001

1. Objectives

To suꢈey toxic dinoflagellate spoꢇes in the ballast tanks of sꢍipꢊ entering Austꢇaliaꢐ poꢇtsꢌ and to develop appꢇopꢇiate control action to protꢋct Austꢇalia's aquacultuꢇe industꢇy.

2. Staff employed on the program

Dr. G. M. Hallꢋgꢇaꢋff (CꢉIRO) Mr. C. J. Bolch (FIRDC)

ꢉupeꢈiꢊoꢇ of projꢋct

Full-time ꢇeꢊꢋarch aꢊꢊiꢊtant

Dinoflagellate culturing

Dr. S. I. Blackbuꢇn (CꢉIRO)

Collaborators

Mr. N. Blainꢌ AGALꢌ Mꢋlbourne

ballaꢊt watꢋꢇ monitoring

Dr. P. Hutcꢍingꢊꢌ

Auꢊtꢇalian Muꢊꢋumꢌ ꢉydnꢋy

biology of ballaꢊt water

Dr. J. Mꢋꢅonꢌ AQIꢉꢌ Canbꢋꢇꢇa

ballaꢊt watꢋꢇ monitoring

ballaꢊt wateꢇ monitoring

dinoflagellate toxin cꢍꢋmiꢊtꢎ

Mꢇ. R. M�rphyꢌ AQIꢉꢌ Canbeꢇꢇa

Dꢇ. Y. Oꢊhimaꢌ

Tohoku Univeꢇꢊityꢌ Japan

Dꢇ. M. Jonꢋꢊꢌ BRRꢌ Canbꢋrra

ballaꢊt watꢋr guidꢋlineꢊ

Dꢇ. G. Rigbyꢌ

BHP Rꢋꢊearch Laboꢇatoriesꢌ Nꢋwcaꢊtle

ship enginꢋꢋring

Mꢇ. C. Scholiꢐꢌ

Woodꢊhole Oceanogꢇaphic Iꢐstitutioꢐ

dinoflagellate genetics

Mꢇ. A. Sꢍeꢏiꢐꢌ AQIꢉꢌ Canbeꢇꢇa

ballast wateꢇ monitoring

Dꢇ. S.O.ꢉtanleyꢌ

Taꢊmanian Dept. of ꢉꢋa Fiꢊhꢋriꢋs

fish farm diꢊꢋaꢊꢋꢊ

Dꢇ. M. Williamsꢌ BRRꢌ Canbeꢇꢇa

ballaꢊt watꢋr guidꢋlineꢊ

Mꢇ. R. Williamsꢌ

NSW Dept. of Fisheꢇiesꢌ Cꢇonulla

biology of ballaꢊt watꢋꢇ

3. Introduction

·ꢀꢁꢂ\

The Australian economy is heavily dependent on the export of raw materials suꢌh as coal, graꢋn, iron ore, wheat and woodchips. Speꢌialised bꢀꢁk ꢌontaꢋneꢈ shꢋps only caꢈꢈy these ꢌommodities ꢋn one diꢈectꢋon (Fꢋg.1), using wateꢈ as ballast foꢈ stabꢋlity on the retuꢈn voyage. As a result, some 60 milꢉꢋon tonnes of baꢉlast wateꢈ aꢈe disꢌhaꢈged in Austꢈalian ports eveꢈy yeꢇꢈ, with ships sailing from Japan acꢌounting for oveꢈ a thiꢈd of this amount (Hutꢌhꢋngs et a/., 1987; Jones, 1991). Caꢈgo vesseꢉ ballast wateꢈ was fiꢈst suggested as a veꢌtoꢈ in the dispeꢈsꢇl of non-indigenous maꢈine sꢆeꢌies

neaꢈly nꢋnety yeaꢈs ago. The diatom Odontella (Biddulphia) sinensis, weꢉꢉ

known fꢈom the tropiꢌal and subtropiꢌal ꢌoasts of the lndo-Pacifiꢌ, was not ꢈepoꢈted ꢋn Euꢈoꢆean waters untiꢉ 1903, when ꢋt pꢈoduꢌed dense plankton blooms in the North Sea. Since it was unlikely thqt tꢅis laꢈge diatom ꢌould

have been oveꢈꢉooked previousꢉy, and ꢋmpossible that it ꢌould have been

ꢌaꢈꢈꢋed by ꢌuꢈꢈents fꢈꢃm distꢇnt oꢌeans, Ostenfeld (1908) suggested thꢇt tꢅis speꢌꢋes was intꢈoduced by sꢅip, eitheꢈ ꢇs paꢈt of the fouꢉing communꢋty on vessels' hulls oꢈ, moꢈe likely, dischaꢈged with the wateꢈ oꢈ sedꢋment ꢌontained ꢋn shiꢆs' ballast tanks. Sinꢌe then, ballast wꢇteꢈ has been suggested as ꢇ disꢆeꢈsal meꢌhꢇnism foꢈ ꢇ lꢇꢈge ꢈꢇnge of maꢈꢋne oꢈganisms (see ꢈeview by Cꢇꢈlton, 1985). In Austꢈꢇlꢋa theꢈe ꢇꢈe at least 15 estꢇblꢋshed maꢈine sꢆeꢌꢋes known with ꢈeasonable ꢌeꢈtainty to hꢇve aꢈꢈived ꢋn ballꢇst wateꢈ. Tꢅese ꢋnꢌlude fish ( 4 sꢆꢆ.), ꢌꢈustꢇꢌeans (4), ꢆoꢉyꢌꢅaete woꢈms (3), moꢉlusꢌs (3) and ꢇ seaweed (1) (Pollꢇꢈd & Hutꢌꢅings, 1990 a, b; see Fꢋg. 2).

Tꢅe ꢆotential imꢆaꢌt of these species on commeꢈꢌial fisheꢈies, aquaꢌultuꢈe and the natuꢈal envꢋꢈonment ꢋs of ꢋnꢌꢈeasꢋng ꢌonꢌeꢈn. In Tasmꢇnia, the intꢈoduꢌed seaweed Undaria pinnatifi da (ꢌuꢈꢈent standing cꢈoꢆ aꢆꢆꢈoximꢇtely 400 tonnes) ꢆoses a thꢈeat to abalone and s�a uꢈꢌꢅin fꢋsꢅeꢈꢋes ꢇs welꢉ ꢇs ꢇ ꢆꢈoposed mꢇꢈine ꢆaꢈk zone (Sꢇndeꢈson, 1990). In the Gꢈeꢇt Lꢇkes of ꢂꢃꢄꢅ Ameꢈꢋꢌa, bꢉoꢌkꢇge of wꢇteꢈ inlets by tꢅe ꢋntꢈoduꢌed zebꢈꢇ musseꢉ Dreissena polymorpha has been estimꢇted to ꢌost mꢃꢈe thꢇn US $ 5 bꢋlliꢃn ꢋn ꢌontꢈꢃl ꢌosts oveꢈ 10 yeꢇꢈs (Hebeꢈt et al., 1989).

If dꢋnofꢉꢇgeꢉꢉꢇte sꢆeꢌꢋes that ꢌꢇn ꢆꢈoduꢌe ꢆꢇꢈꢇꢉꢊꢋꢌ shellfish toxꢋns
(FIRTA gꢈꢇnt 86 I 84) aꢈe ꢋntꢈoduꢌed into sensitive ꢇquaꢌultuꢈe aꢈeꢇs, tꢅe ꢈesults ꢌould be disastꢈous foꢈ ꢌommeꢈꢌiꢇl shellfish faꢈming. The ꢆꢈesent

Fig.1 . Introduction of non-endemic, toxic dinoflagellate cysts via ships' ballast water, and their potential impacf on Australian aquaculture.

OVERSEAS PORT

Bulk cargo unloaded

from holds

1n harꢀur surface sediment

Vessel takes on water into ballast tanks and hold including planktonic and resuspended cysts

ꢌ;

Dinoflagellate bloom in water including cysts

ꢁꢂꢃ

ꢄꢅꢆ

Resuspended sediment and cysts

AUSTRALIAN PORT

g

n

cludin

poꢀ in

loading

out into

t pumped

Ballas

Cysts seed dinoflagellate blooms in the ꢁꢂꢀ el the

Vess

isms.

organ

other

sts and

any cy

when conditions are suitable

t.

expor

argo for

bulk c

loads

Blꢅꢅm kills farmed finfish

�

ꢄ

form new

inate to

Cysts germ

transferred cysts seꢁꢂe to the ꢃꢄꢅm

study was pꢇomꢖtꢋd by tꢍe ꢇꢋꢄꢋꢐt appꢋaꢇaꢐꢄꢋ iꢐ Australiaꢐ watꢋrs of tꢗxꢃꢄ diꢐoflagellate spꢋꢄiꢋs tꢍat aꢖꢖꢋaꢇ to have a disjuꢐꢄt global distrꢃbutꢃoꢐ, suꢄꢍ as Gymnodinium catenatum (ꢄoꢐfiꢐed to Japaꢐꢌ Mꢋxiꢄo, soutꢍꢋrꢐ Eurꢗꢖꢋ aꢐd Tasmaꢐia), Alexandrium cateneꢃa ꢔcoꢐfiꢐed to North Amꢋrꢃꢄa,Japan, ꢀouth Afꢇiꢄa aꢐd soutꢍꢋast Austꢇalia) aꢐd Alexandrium minutum (ꢄoꢐfinꢋd to ꢀouth Australia aꢐd soutꢍꢋꢇꢐ Euꢇopꢋ) ꢔsꢋꢋ Fig. 3a,b). ꢀtrꢗꢐg ꢄiꢇꢄumstaꢐtial ꢋvidꢋꢐꢄꢋ provꢃdꢋd by ꢋxamiꢐatioꢐ of ꢍꢃstꢗrꢃꢄ ꢖlankton samplꢋs ꢔHallꢋgraꢋff et al.,1988), ꢄyst suꢑꢋys iꢐ sꢋdꢃmꢋꢐt dꢋꢖtꢍ ꢄorꢋs (Bolch & Hallꢋgraꢋff, 1990) aꢐd gꢋꢐꢋtꢃc studꢃꢋs uꢊꢃꢐg ꢋꢐzyme ꢋlꢋꢄtropꢍorꢋsis and sꢋxual ꢄomꢖatꢃbꢃlꢃty ꢋxꢖꢋrimꢋꢐts (Biaꢄkburꢐ ꢋt al.,1 989) suggꢋstꢋd tꢍat tꢍꢋ toxic dꢃꢐoflagꢋllatꢋ Gymnodinium catenatum waꢊ iꢐtꢇoduꢄꢋd to Tasmaꢐꢃa at somꢋ tꢃmꢋ aftꢋꢇ 1980 iꢐ tꢍe arꢋa arouꢐd tꢍꢋ sꢍiꢖpiꢐg port of Hobart. Bꢋntꢍꢃꢄ ꢄyst bꢋds of tꢍꢃs sꢖꢋꢄꢃꢋꢊ arꢋ now wꢃdꢋsꢖrꢋad iꢐ sꢗutꢍꢋrꢐ Taꢊmanꢃa, aꢐd dꢋꢐsꢋ ꢖlaꢐktꢗn bloomꢊ ꢃn 1 986 and 1991 iꢐ soutꢍꢋrꢐ Taꢊmanꢃaꢐ watꢋrs ꢐꢋꢄꢋssꢃtatꢋd tꢍꢋ ꢄlꢗꢊurꢋ ꢗf 1 5 ꢊꢍellfꢃꢊꢍ farms foꢇ ꢖꢋriods of uꢖ to 6 mꢗntꢍs (Hallꢋgraꢋff & ꢀumnꢋr, 1 986). ꢀꢃmꢃlarly, ꢇRNA sꢋquꢋꢐciꢐg ꢍas sꢍowꢐ a ꢇꢋmarkablꢋ matꢄꢍ bꢋtwꢋꢋꢐ Auꢊtralꢃaꢐ (ꢒoꢓ ꢕꢍꢃllip Bay) aꢐd Jaꢖaꢐꢋsꢋ ꢖoꢖulatioꢐs of tꢍꢋ toxꢃꢄ dꢃnꢗflagellate Alexandrium catenella, aꢐd bꢋtweꢋꢐ Australiaꢐ ꢔꢕoꢅ Rꢃvꢋr, Adelaꢃdꢋ) aꢐd Euꢇoꢖꢋaꢐ poꢖulatꢃoꢐs of tꢍꢋ toxꢃꢄ diꢐoflagꢋllatꢋ A. minutum (ꢀꢄꢍꢗlin & Aꢐdꢋꢇsoꢐꢌ 1991).

Thꢋ aims of tꢍꢋ prꢋsꢋnt work wꢋrꢋ (1) to ꢋstablꢃsꢍ unambꢃguouꢊly wꢍꢋtꢍꢋꢇ toxiꢄ dꢃꢐoflagꢋllatꢋ sꢖꢋꢄiꢋs ꢄaꢐ bꢋ ꢄarriꢋd by sꢍꢃpꢊ ꢃn a vꢃablꢋ ꢄoꢐditioꢐ; (2) to ꢋluꢄꢃdatꢋ ꢍow dꢃꢐoflagꢋllatꢋ ꢄyst abundaꢐꢄꢋ ꢃn ballast tanks

varꢃꢋs aꢄꢄordꢃꢐg to sꢍꢃꢖ typꢋ, ballast-taꢐk ꢄoꢐfꢃguratꢃoꢐ, sꢋaꢊꢗꢐ and ꢖꢗꢘ of oꢇꢃgiꢐ, aꢐd tꢍꢋ pꢇaꢄtꢃꢄꢋ of mꢃd-oꢄꢋaꢐ ballast ꢋxꢄꢍaꢐgꢋ; aꢐd (3) to ꢋxaminꢋ the efficaꢄy of vaꢇious physiꢄal aꢐd chꢋmiꢄal trꢋatmꢋꢐts to kꢃll tꢗxꢃꢄ diꢐoflagꢋllate ꢄysts iꢐ ballast watꢋꢇ samplꢋs.

4. Ballast water survey

4.1 Materials and methods

QIꢀ)

ioꢐ ꢀꢋꢙꢃꢄꢋ (A

raꢐtiꢐe aꢐd Iꢐspꢋꢄt

Austꢇaliaꢐ Qua

Offiꢄꢋꢇs of tꢍꢋ

wꢃdely

aꢇrivꢋd iꢐ 18

vessꢋls wꢍiꢄh

of 343 caꢇgo

ballast taꢐks

samplꢋd tꢍe spꢇꢋad Australiaꢐ

. 4).

Marꢄh 1990 (Fig

vembeꢇ 1987 aꢐd

poꢅs bꢋtwꢋꢋꢐ No

saꢃlꢋd fꢇom J

frꢗm

aiꢐdꢋr arrꢃvꢋd

apaꢐ; thꢋ ꢇꢋm

f tꢍꢋ sꢍips had

Moꢇe thaꢐ hal

rꢋ, ꢀri

h Koꢇꢋa, ꢀꢃngaꢖo

, Iꢐdoꢐꢋsia, ꢀout

aꢐd Hoꢐg Koꢐg

ꢋast Asia (Cꢍiꢐa

Fig.2. Suspected ballast water introductions of marine organisms into Australia

Aeolidiella indica (Japan) Musculista senhousia (Asia) Theora Jubrica (Japan)

Molluscs

Eurylana arcuata (New Zealand)

Neomysis japonica (Japan)

Pyromaia tuberculata (?New Zealand) Tanais dulongi (Europe)

Boccardia proboscoidea (?Japan) Mercierella enigmatica {fndia) Pseudopolydora paucibranchiata (?Japan)

Annelida

M

Acanthogobꢑuꢓ ꢐavꢑꢒanuꢓ (Japan) Lateobrax japonꢑcuꢓ (Japan)

Sparindentex hasta (India)

Tridentiger trigonocephalus (Japan)

Fig.3a. Global sꢀreading of toxic dinoflagellate bloo0ms (ꢀaralytic shellfish ꢀoisoning ) between 1970 and 1 99 .

Fig.3b. Toxic dinoflagellate blooms first aꢀꢀeared in Australian estuaries in the late 1980s, caused by Gymnodinium

catenatum (g) in the ꢀoꢁ of Hobart, Alexandrium

minutum (m) in the ꢂoꢃ of Adelaide, and Alexandrium cateꢀꢁꢂa (c) in the ꢀoꢃ of Melbourne. The disjunct global distribution of these species raised susꢀicions about ballast water as a ꢀossible disꢀersal mechanism.

LaꢐkaꢌTaiwaꢐ)ꢌ tꢍꢋ Mꢃddlꢋ East (Turkꢋyꢌ Iraꢐꢌ Yꢋmꢋꢐ)ꢌ Nortꢍ Amerꢃca (Caꢐada aꢐd tꢍꢋ Unitꢋd ꢀtatꢋs)ꢌ Argꢋntꢃnaꢌ Mꢋxico and Nꢋw Zꢋaland. After ꢖartial ꢇꢋmoval of ballast watꢋr in Australꢃan ꢖoꢅsꢌ aꢄcess was gaꢃned to tꢍe ballast tanks oꢇ ꢍolds of vꢋssꢋls. Rꢋꢖlꢃcatꢋ 500 mL samꢖlꢋs of ballast-tank sꢋdimꢋꢐt orꢌ if absꢋnt, ballast watꢋꢇ samꢖlꢋs wꢋꢇꢋ collꢋctꢋd by ꢍand from diffꢋꢇꢋꢐt arꢋas of tꢍꢋ vꢋssꢋls. ꢀamꢖlꢋs wꢋrꢋ storꢋd iꢐ a dark ꢇꢋfrꢃgerator (4 oc) uꢐtil dꢋsꢖatcꢍꢋd to CꢀIRO for analysis.

Ballast-tank samꢖlꢋs wꢋrꢋ sonꢃcatꢋd for 2 min (Braun Labsoniꢄ
ꢍomogꢋnisꢋrꢌ 1 00 W) to dꢃslodge detrꢃtus ꢖartꢃcles. ꢀamꢖles were tꢍen fractionated tꢍrougꢍ a 90 uꢚ sꢃꢋvꢋ onto a 20 uꢚ sꢃevꢋ. Tꢍꢃs concentrated matꢋrial was ꢋxamꢃnꢋd by ligꢍt and sꢄannꢃng ꢋlꢋctron mꢃcroscoꢖy. Viable dꢃꢐoflagꢋllatꢋ cysts wꢋrꢋ isolatꢋd by mꢃcroꢖꢃꢖꢋttꢋ and wasꢍꢋd sevꢋral times ꢃn stꢋrilꢋ culturꢋ medꢃum. Tꢍꢋ medꢃum consꢃstꢋd of fꢃltꢋrꢋd seawater (28 Ofoo salꢃnꢃty) autꢗclaved ꢃn tꢋflon ꢄontaꢃnꢋrsꢌ wꢃtꢍ nutrꢃꢋnts added accꢗrding to Loꢋblicꢍ's (1 975) GꢕM mꢋdꢃum modꢃfꢃꢋd wꢃtꢍ sꢋlꢋnꢃum (H2ꢀe03 at 1 o-8

M fꢃnal concꢋntratꢃꢗn).ꢀꢃnglꢋ cysts wꢋrꢋ ꢖlacꢋd in sterꢃlꢋ ꢖolystyrene petrꢃ dꢃsꢍꢋs (36 mm dꢃameter) wꢍꢃcꢍ contaꢃnꢋd 5 mL mꢋdꢃum. Tꢍꢋ cysts were ꢃncubatꢋd at 18 oc (1 2:1 2 ꢍ lꢃgꢍt: dark) at aꢐ irradꢃance of 80 uE m-2s-1 and wꢋrꢋ ꢋxamꢃnꢋd daily for gꢋrmꢃnatꢃon.

4.2. Results of ballast water survey

Of tꢍꢋ total of 343 ballast-tank samꢖles ꢋxamined, 65% contaꢃned fꢃne brown oꢇ black sedꢃment (uꢖ to an ꢋstꢃmatꢋd 1 00 tonnꢋs of sedꢃmꢋnt ꢖer sꢍiꢖ). ꢀomꢋ of tꢍꢋ remaꢃnꢃng samꢖlꢋs werꢋ sꢋdꢃment-freꢋ becauꢊe tꢍe bottoms of tꢍꢋ ballast tanks werꢋ ꢐot ꢋasꢃly accꢋssiblꢋ for samꢖlꢃng. One ꢍuꢐdrꢋd samꢖlꢋs of sedꢃmꢋnt from 1 8 Australꢃaꢐ ꢖoꢅs wꢋrꢋ selected for dꢋtaꢃlꢋd study. Tꢍꢋ cꢋllulosꢋ tꢍꢋcaꢋ of motꢃlꢋ dꢃꢐoflagꢋllatꢋs wꢋrꢋ rarely sꢋꢋꢐ iꢐ ballast-tank samꢖlꢋsꢌ but ꢇꢋstiꢐg cysts wꢋꢇꢋ commoꢐ. Dꢃnoflagꢋllatꢋ cysts were fouꢐd in samꢖlꢋs from vꢋssꢋls eꢐtꢋriꢐg all 18 Australiaꢐ ꢖorts studiꢋd (Fig. 4). Tꢍꢋ cysts wꢋrꢋ found in 50 of tꢍꢋ 1 00 sꢋdꢃmꢋnt contaꢃning samꢖlꢋs; 53 sꢖꢋciꢋs of cysts wꢋrꢋ idꢋntifꢃꢋdꢌ of wꢍꢃꢄꢍ 20 sꢖꢋcꢃꢋs werꢋ gꢋrminatꢋd to ꢖroduꢄꢋ viablꢋ culturꢋs (Tablꢋ 1). Tꢍus tꢍꢋ rꢃsk of foreign micro-organisms bꢋꢃꢐg iꢐtꢇoduꢄꢋd iꢐto Australꢃaꢐ watꢋrs vꢃa sꢍꢃꢖs' ballast watꢋr is wꢃdꢋsꢖrꢋad aꢐd tꢍꢋ iꢐoculum ꢇꢋꢖrꢋsꢋꢐts a grꢋat dꢃvꢋrsꢃty of sꢖꢋciꢋs.

Tꢍe idꢋꢐtifꢃcation of cysts of tꢍꢋ toxic diꢐoflagꢋllatꢋs Alexandrium catenel/a aꢐd A. tamarense ꢃn 5 of tꢍꢋ 100 sꢋdimꢋnt samꢖlꢋs was of

'

Australia

0

�

Poꢅ Lꢆꢇꢆ

nna

abu

ri

T

Fig. 4. Maꢀ of Australia showing the location of the 18 ꢀorts from which ballast-tank samꢀles were collected. Samꢀles from 9 ꢀorts contained toxic dinoflagel late cysts

(fi lled squares).

Fig.Sa. Live cysts of the toxic di noflagel late Alexaꢀdrium tamareꢀse from a vessel arriving in Eden from Muroran,

Japan.

•

Fig.Sb. Motile cells of the toxic di noflagel late Alexandrium catꢁꢀꢁlla, cultured by germi nating ballast water cysts

from a vessel arriving in Port Hedland from Kashima,

Jaꢀan.

Tabꢋe
1. Diꢇofꢋageꢋꢋ ate cy

ꢄamꢅꢋeꢄ (53 ꢄꢅꢆi
ꢄt ꢄꢀec

ieꢄ in

baꢋꢋaꢄt

tank

20 ꢄꢀꢀ. cꢊꢋtꢊꢌꢍꢎꢏ eꢄ· ide-ꢇꢈifiꢉ;

Alꢗxꢚꢝdriuꢛ ꢔꢕꢖꢗꢘꢗllꢙꢚꢛꢚꢜꢗꢝꢞꢗ

Diplopelta

Diplopsalis

Diplopsalopsis

^Gonyâûlâ^x(7 ^spp)

Gymnodinium catenatum Lingulodinium

Peridinium

Pheopolykrikos

Polykrikos

Protoceratium

Protoperidinium (22 spp) Pyrocystis

^{Scrippsꢀeꢁa}(6 ^s^p^p⁾

Zygabikodinium

^Ta^b^lê2. Ôꢌigiⁿôf20 ^bâꢋꢋaꢄt^t^{aꢇk ꢄamꢀꢋeꢄ}^côꢇ^tâiⁿ^{iꢇg toxi}^c

diꢇoflageꢋꢋate cyꢄtꢄ.

KOREA

JAPAN

Chiba (Aiexandrium sp.) Fukuyama (G.catenatum) Kashima (A. catenella) Kure (Aiexandrium sp.)

Kushiro (Aiexandrium sp.) Muroran (A.tamarense) Shimizu (A.tamarense)

U be ( G. catenatum)

lnchon (G. catenatum)

Kohong (G.catenatum) Kwangyang (Aiexandrium) Pohang (Aiexandrium sp.) Samchonpo(A.tamarense)

coꢐsidꢋꢇablꢋ coꢐcꢋꢇꢐ (Fꢃg. Saꢌ b). Aftꢋꢇ tꢍis CꢀIRO suꢙꢋy was comꢖletꢋd, tꢍꢋ Austꢇaliaꢐ Quaꢛaꢐtꢃꢐꢋ aꢐd Iꢐsꢖꢋctꢃoꢐ ꢀꢋꢇvꢃcꢋ (AQIꢀ) contractꢋd tꢍꢋ Austꢇaliaꢐ Govꢋꢛꢐmꢋnt Aꢐalytꢃcal Laboꢛatoꢇiꢋs ꢔAGAL) to contꢃnuꢋ moꢐitoꢇiꢐg tꢍꢋ ballast taꢐks of sꢍꢃꢖs ꢋꢐtꢋꢇiꢐg Austꢛaliaꢐ ꢖoꢛts. Tꢍꢃs moꢐitoꢇiꢐg dꢋtꢋctꢋd 9 fuꢛtꢍꢋꢇ ꢖosꢃtivꢋ Alexandrium ballast-taꢐk samꢖlꢋs. In 5 casꢋsꢌ tꢍꢋs� Alexandrium cysts ꢍavꢋ bꢋꢋꢐ succꢋssfully gꢋꢛmꢃnatꢋd ꢃnto viablꢋ diꢐoflagꢋllatꢋ cultuꢛꢋs and tꢍꢋiꢇ toxicꢃty ꢍas bꢋꢋꢐ coꢐfꢃꢛmꢋd by ꢍigꢍ- ꢖꢋꢛfoꢇmaꢐcꢋ lꢃquꢃd cꢍromatogꢛaꢖꢍy ( Osꢍima, Bolcꢍ & Hallꢋgꢛaꢋffꢌ iꢐ ꢖress). Fouꢇ samꢖlꢋs coꢐtaiꢐꢃꢐg toxꢃc Gymnodinium catenatum cysts wꢋrꢋ also ꢃdꢋꢐtifiꢋd. Toxic diꢐoflagꢋllatꢋ cysts wꢋꢛꢋ fouꢐd iꢐ woodcꢍꢃꢖ, gas and oꢛe vꢋssꢋlsꢌ wꢍicꢍ saꢃlꢋd fꢛom ꢋꢃtꢍꢋꢇ Jaꢖaꢐ oꢇ ꢀoutꢍ Koꢛꢋa and ꢋntꢋred 9 Austꢇaliaꢐ ꢖoꢛts (Tablꢋ 2). Onꢋ ballast taꢐk was fꢃllꢋd durꢃng a toxꢃc dꢃꢐoflagꢋllatꢋ bloom iꢐ tꢍꢋ ꢖoꢅ of Muꢛoꢛaꢐ, Jaꢖaꢐ, durꢃng July 1 989. A samꢖlꢋ takꢋꢐ iꢐ Edꢋꢐ, Austꢛalꢃa, was ꢋstꢃmatꢋd to contaꢃn morꢋ tꢍan 300 mꢃllꢃoꢐ Alexandrium cysts. Onꢋ montꢍ latꢋꢇꢌ oꢐ ꢃts nꢋxt voyagꢋ from ꢀꢍimizu,Jaꢖaꢐꢌ to Edꢋꢐꢌ tꢍꢃs sꢍꢃꢖ stꢃll coꢐtaꢃꢐꢋd tꢍꢋ samꢋ cyst materꢃal, albꢋꢃt iꢐ lowꢋꢇ· coꢐcꢋꢐtꢛatꢃoꢐ. Iꢐ botꢍ casꢋs tꢍꢋsꢋ cysts faꢃlꢋd to gꢋꢛmꢃnate uꢐtꢃl bout 6 moꢐtꢍs latꢋꢇꢌ wꢍꢃcꢍ suggꢋsts tꢍat tꢍꢋy wꢋrꢋ ꢐꢋwly foꢛmꢋd cysts uꢐdꢋꢇgoꢃꢐg a maꢐdatoꢛy dormaꢐcy ꢖꢋꢛꢃod ꢔAꢐdꢋꢛson, 1 980) ꢛatꢍꢋꢛ than matuꢛꢋ cysts ꢇꢋsusꢖꢋꢐdꢋd from ꢍarbouꢛ sꢋdꢃmꢋnts durꢃng sꢍiꢖs' ballasting. Analysꢃs of vꢋssꢋls' dꢋꢖaꢅuꢛꢋ datꢋs ꢔmoꢛꢋ lꢃvꢋ dꢃꢐoflagellatꢋ cysts from May to Octobeꢛ) suꢖꢖoꢛts tꢍꢋ vꢃꢋw tꢍat tꢍꢋ watꢋꢛ columꢐ durꢃng seasonal (sꢖring to ꢋarly autumꢐ) dꢃnolagꢋllatꢋ blooms is tꢍꢋ ꢖꢛꢋdomꢃnant orꢃgꢃn of ballastwatꢋꢇ cysts.

4.3. Conclusions of ballast water survey

Tꢍꢋ ꢖꢇꢋsꢋꢐt work ꢋstablꢃsꢍꢋd uꢐambꢃguously tꢍat toxꢃc dꢃꢐoflagꢋllatꢋs ꢌ ꢋsꢖꢋcꢃally tꢍosꢋ sꢖꢋcꢃꢋs tꢍat ꢖꢇoducꢋ ꢇꢋsꢃstant ꢇꢋsting cysts, caꢐ bꢋ traꢐsꢖortꢋd iꢐ a viablꢋ form iꢐ tꢍꢋ ballast watꢋꢛ of cargo vꢋssꢋls. Tꢍꢋ Austꢇaliaꢐ Quarantꢃꢐꢋ aꢐd Iꢐsꢖꢋctꢃoꢐ ꢀꢋrvꢃcꢋ ꢍas ꢇꢋsꢖonded to tꢍꢋ ꢖꢇꢋcꢋdiꢐg ꢋvidꢋꢐcꢋ by declaꢛꢃꢐg tꢍꢋ dꢃsꢖosal of ballast watꢋꢛ to be a quaꢇaꢐtiꢐꢋ issuꢋ of ꢐatioꢐal sꢃgꢐꢃficaꢐcꢋ. As of 1 Fꢋbruaꢇy 1 990, Austꢛalꢃa iꢐtꢇoducꢋd voluꢐtaꢇy guꢃdꢋlꢃꢐes foꢇ sꢍꢃꢖs ꢋꢐtꢋꢇꢃꢐg ꢃts ꢖoꢛts from ovꢋꢛseas (Fꢃg. 6).Tꢍꢋsꢋ mꢋasurꢋs iꢐcludꢋ tꢍꢋ sꢍꢃꢖs ꢖꢛovꢃdꢃꢐg cꢋꢛtꢃfꢃcatꢃoꢐ tꢍat watꢋꢛ aꢐd sꢋdimꢋꢐt iꢐ tꢍꢋ ballastꢃng ꢖoꢛt aꢛꢋ fꢛꢋꢋ fꢛom ꢍaꢛmful oꢛgaꢐꢃsms (now ꢇꢋgulaꢇly usꢋd by somꢋ sꢍiꢖs orꢃgꢃꢐatꢃꢐg fꢇom Nꢋw Zꢋaland), ꢇꢋ-ballastꢃng ꢃꢐ mꢃd-ocꢋaꢐ (now ꢇꢋgularly used by woodcꢍꢃꢖ caꢛgo vꢋssꢋls sꢋꢛvꢃcꢃng

ꢈꢉꢊꢋꢌꢍꢎꢏ1ꢐꢑ Ouaranti�e and inspection Serv1ce

Dꢓꢔꢕꢖꢗꢘꢙꢚ ^o;

lncustr:es and Energy

�-�ꢟꢠꢡꢢ

Ballast Water

a serious quarantine problem

Fig.6.

Ballast water leaflet issued by the Australian Quarantine

âⁿ^dÎn^spectⁱ^{on Serv}ⁱ^{ce o}ⁿ¹^Fêbrûa^ry1990.

TꢛiabuꢐꢐaꢌTasmaꢐia)ꢌ maꢣiꢐg a cꢢmmꢜtmꢋꢐt ꢐꢢt tꢢ dꢜscꢍaꢛgꢋ ballast oꢛ tꢢ ꢣꢋꢋp ballast taꢐꢣs as clꢋaꢐ as pꢢssꢜblꢋ. As ꢢf 1 Nꢢvꢋmbꢋꢛ 1991ꢌ tꢍꢋ Maꢛiꢐꢋ Eꢐviꢛꢢꢐmꢋꢐt Pꢢllutiꢢꢐ Cꢢmmꢜttꢋꢋ (MEPC) ꢢf tꢍꢋ Intꢋꢛꢐatꢜꢢꢐal Maꢛꢃꢞimꢋ Oꢛgaꢐisatiꢢn (IMO) ꢍas ratꢜfꢜꢋd tꢍꢋ abꢢvꢋ guidꢋlꢜꢐꢋs fꢢꢛ adꢢꢖtꢃon ꢢn aꢐ iꢐtꢋꢛꢐatꢜꢢꢐal basꢜs.

A majꢢꢛ pꢛꢢblꢋm wꢜtꢍ mꢜdꢢcꢋaꢐ ꢋxcꢍaꢐgꢋ ꢢf ballast watꢋꢛ ꢜs tꢍat ꢜt can
ꢢnly bꢋ caꢛꢛied ꢢut by vꢋssꢋls up tꢢ 40ꢌ000 dꢋad wꢋꢃgꢍt tꢢnnagꢋ (such as wꢢꢢdcꢍip caꢛgꢢ vꢋssꢋls)ꢌ aꢐd ꢐꢢt by tꢍꢋ majꢢꢛity ꢢf tꢍꢋ laꢛgꢋ ꢢꢛꢋꢠcaꢛgo vꢋssꢋls vꢜsꢜtiꢐg tꢍꢋ Austꢛalꢜaꢐ pꢢꢛts ꢢf Pꢢꢛt Hꢋdlaꢐdꢌ Damꢖꢜꢋꢛ aꢐd Gladstꢢꢐꢋ. Fuꢛtꢍꢋꢛmꢢꢛꢋꢌ wꢍꢜlꢋ mꢜdꢢcꢋaꢐ ꢋxcꢍaꢐgꢋ ꢢf ballast watꢋꢛs may bꢋ ꢋffꢋctꢜvꢋ iꢐ ꢛꢋmꢢvꢜꢐg ꢢꢛgaꢐꢜsms iꢐ tꢍꢋ watꢋꢛ cꢢlumꢐ ꢢf ballast tanksꢌ it ꢜs ꢢꢐly paꢅꢜally ꢋffꢋctꢃvꢋ iꢐ ꢛꢋmꢢvꢜng dꢜnꢢflagꢋllatꢋ cysts wꢍꢜcꢍ ꢍavꢋ sꢋtꢞlꢋd to tꢍꢋ bꢢttꢢm ꢢf ballast tanks. Among 32 vꢋssꢋls wꢍꢃcꢍ ꢋxꢖlꢃcꢃtly claimed to ꢍavꢋ ꢋxcꢍangꢋd ballast watꢋꢛ ꢜn mꢃdꢠꢢcꢋaꢐꢌ 1 4 wꢋꢛꢋ still found to conꢞaꢃn sꢜgꢐꢜfꢜcaꢐt amꢢunts ꢢf sꢋdꢃmꢋntsꢌ ꢜncludꢜng dꢃꢐoflagꢋllatꢋ cysts. A mꢗꢛꢋ ꢋffꢋctivꢋ mꢋasuꢛꢋ wꢢuld bꢋ to avꢢꢃd ballastꢜng duꢛꢃng toxꢃc dꢃnoflagꢋllatꢋ blꢢꢢms iꢐ fꢢꢛꢋigꢐ ꢖꢢꢛts.

5. Ballast water exchange study on M.V."Iron Whyalla"

Tꢍꢋ ꢋffꢋctꢃvꢋnꢋss of ꢛꢋballastꢃng oꢖꢋꢛatꢃons ꢃn mꢃdꢠocꢋan was sꢞudied ꢃn dꢋtaꢃl ꢢꢐ.bꢢaꢛd tꢍꢋ BHPꢠꢢwnꢋd vꢋssꢋl "Iꢛon Wꢍyalla"ꢌ ꢋn ꢛꢢuꢞꢋ fꢛom ꢀꢃꢐgaꢖꢢꢛꢋ tꢢ Pꢢꢛt Hꢋdland. Tꢍꢋ "Iꢛꢢꢐ Wꢍyalla ꢜs a nꢃnꢋ caꢛgꢢ ꢍold sꢃnglꢋ- dꢋcꢣ bulꢣ caꢛꢛꢜꢋꢛ wꢃtꢍ a tꢢtal ballast caꢖacꢃty ꢢf 76,330 tonnꢋs (Fꢃg. 7a). Fꢢllꢢwꢜng a cꢢmꢖlꢋtꢋ clꢋanꢠꢢut of ballast tanꢣs ꢜn dꢎ dockꢌ tꢍꢋ sꢍꢃꢖ took in ballast watꢋꢛ ꢜn ꢀꢃngaꢖoꢛꢋ Haꢛbouꢛ ꢢn 13 and 1 4 ꢀꢋꢖt 1 991 . Condiꢞions duꢛꢜng ballastꢃng wꢋꢛꢋ watꢋꢛ tꢋmꢖꢋꢛatuꢛꢋ 30 ꢢcꢌ salꢃnꢃty 25 Ofoo (bꢗꢞꢍ ꢞaken at 1Om dꢋꢖtꢍ) ꢌtꢍꢋ ꢜncꢜdꢋncꢋ ꢢf a dꢋnsꢋ (1 o7 cꢋlls/lꢃtꢋꢛ) dꢃatom bloom (maꢜnly Chaetoceros,Skeletonema) and sꢖaꢛsꢋ ꢐꢢnꢠtoxꢃc dꢃnoflagꢋllatꢋ

ꢖlaꢐꢣtꢢꢐ (Ceratium, Dinophysis, Diplopsalis, Protoperidinium ). Rꢋsusꢖꢋnded

ꢍaꢛbꢢuꢛ sꢋdꢜmꢋnts wꢋꢛꢋ clꢋaꢛly vꢃsꢃblꢋ wꢍꢋꢐ tꢍꢋ sꢍꢃꢖ lꢋft tꢍꢋ dock on 1 5 ꢀꢋꢖtꢌ1Oamꢌ but wꢋꢛꢋ ꢐot ꢋvꢜdꢋnt duꢛꢃng tꢍꢋ ballastꢜng ꢖꢛocꢋduꢛꢋs (ꢍaꢛbouꢛ dꢋꢖꢞꢍ was 1 3m). Daꢃly obsꢋꢝaꢞꢃons wꢋꢛꢋ madꢋ wꢃtꢍ a mꢃcꢛoscoꢖꢋ on boaꢛd sꢍꢃꢖ tꢢ study tꢍꢋ ꢋffꢋct of ꢋnclosing a natuꢛal communꢃty of ꢖꢍꢟoꢠ and zoꢢꢖlanktꢢn ꢜꢐ a sꢋlꢋctꢋd ballast tank (sꢞaꢛboaꢛd tank no.2ꢌ aꢖꢖꢛꢗximaꢞely 7,000 tꢢnnꢋs caꢖacity). Watꢋꢛ samꢖlꢋs wꢋꢛꢋ wꢃtꢍdꢛawn by mꢋans of a small ꢖꢋꢛꢜstaltꢃc ꢖumꢖ usꢃng sꢃlꢃcꢢꢐ tubꢃng wꢍꢃcꢍ ꢍad bꢋꢋn ꢖꢛꢋꢖosꢃtꢃonꢋd at thꢛee dꢃffꢋꢛꢋꢐt lꢋvꢋls ꢃꢐ tꢍꢋ tanꢣ duꢛꢃng tꢍꢋ dꢡꢠdꢢcꢣ pꢋꢛꢜꢢd. Pꢍysꢜcoꢠcꢍꢋmꢃcal

. Fig. 7a. Ballast tank configuration on thꢔ M. V."Iꢌon Whyaꢋla".

For•p•ak

tank

ꢀ.ꢁ mꢂlnhꢃ'•

aocaaa

Foreward