SURFACE PREPARATION AND APPLICATION OF FOUL-RELEASECOATINGS ON SUBIIERGED COOLING WATER STRUCTURES

David B. Innis

TENERA Environmental Services

P.O. Box 400 Avila Beach, CA 93424 U.S.A.dinnis@teneraenergy.corn

INTRODUCTION

Biofouling Contro] Backgrounct

Electric generation at power facilities across the United States is frequently interrupted to control or removenuisance species that proliferate in cooling water systems. Plumbing systems of both coastal and inlandfacilities, which pump huge quantities of water to remove waste heat provide suitable habitat formicroscopic larvae entrained in cooling water, This same water provides the growing fouling community arich source of food and an outlet for viable larvae that can later re-enter the plumbing to perpetuate thecolonies or establish new ones. Uncontrolled, these pests can quickly block flow in piping or detach andhinder heat exchange, Typical controls range from massive injection of chemicals, such as chlorine,laborious manual scraping, recycling of waste heat, to use of toxic coatings. To overcome the deleteriousside-sects of many of these methods, new techniques evolve to meet environmental, regulatory, andeconomic concerns. Control methods also improve to maintain the availability and generation capacity ofthese facilities. Silicone foul-release coatings applied to settlement surfaces are one new strategy to controlthe proliferation of biofouling in water systems.

The following work represents an interdisciplinary approach to managing the biofouling community thatdevelops within a cooling water system. The management or "control technologies" rely on understandingthe biological and mechanical systems, and their interactions in the power production process. Thesestrategies blend the concerns of commercial operation and regulatory compliance. One of the tools used tominimi7< the risks associated with an oceanic cooling water source is the application of foul-releasecoatings to submerged structures

Diablo Canyon Power Plant Cooling Water System

Diablo Canyon Power Plant DCPP! is a two unit, 2,200 megawatt, nuclear-fueled, P%R power plantoperated by Pacific Gas and Electric Company PG&E!. The plant has operated since 1985 and dischargesapproximately 2.5 billion gallons of cooling water per day under full two-unit operation. It is located on thecentral California coast midway between Los Angeles and San Francisco near San Luis Obispo, CA.

Diablo's cooling water is pumped in from an 80 m �50 ft! wide shoreline intake though conduits to asingle pass open loop condenser. Seawater discharges +12'C +20'F! above ambient that averages 13'C�5'F! and ranges between 10 to 17 C �9 to 60'F; except during El Nino years to 20'C, 68'F!. Ecologistsdescribe the rocky coastline as highly productive with expanses of kelp visible fram shore. The setting isunique, in a practically undisturbed portion of California.

sessile marine animals. On these walls organismsexchanger has little protection from debris that may

are relatively frelater detach

Freshwater steam is condensed back to water by transferring heat to the seawater. Each condenser coolingsystem is served by two main seawater pumps CWP!. The CWPs produce a combined rated flow for eachunit of between 778,000 gpm and 895,000 gpm two-unit combined flow is 1,589,000 gpm nunimum and1,749,000 gpm maximum!. Seawater flow within the conduits ranges between 6 to 7 feet per second. Fromthe condenser the warmed seawater for each unit is then discharged back into the ocean at the shoreline ofDiablo Cove.

The plant's cooling water system conduits present an enormous surface area for the settlement and growthof marine fouling organisms. Two types of fouling occur in a power plant. Micro-fouling by smallunicellular algae and bacteria is a problem in the plant's condenser tube system, where the resulting surfacelayer of fouling organisms reduces the efficiency of heat transfer. Macro-fouling, primarily barnacles andmussels, attach and grow on most untreated surfaces that are exposed to the rapid flow of seawater throughthe plant.

Macro-fouling organisms are filter feeders and the continual flow of seawater through the system providesa rich food source for rapid growth. Left uncontrolled, fast growth and resultant loose shells affect plantefficiency within 6 to 7 months of initial pump operation. This creates the need for cleaning.

At the left, an example of the dominant nuisance species at Diablo is Megabalanus ealifornreus, an acornbarnacle. It prefers to settle on congeners and is cued by chemicals in the shells live, dead, base plates too;Figure 2!. Within the artificial environment created at DCPP, Megabalanus grows crowded, closelyassociated, and rapidly in these conditions. They easily grow to 4 cm �.5 inch! base in 6 months In the

Figure 1 Overview of Diablo Canyon Power Plant andprominent cooling water system components.

DCPP units 1 and 2 have

independent cooling systems for re-condensing freshwater steam for theturbine power cycle. Each unit hasits own system of intake anddischarge conduits, but they allshare the same intake structure and

discharge locations Figure 1!.During normal operation, seawateris drawn from the Intake Cove and

pumped approximately 26 m 85 ft!above sea level fo two condenser

systems through four long conduits.At the intake, submerged bar racksand traveling water screens entrapkelp, seaweed, and larger debris 0 mm; 3/8 iri!. Microscopiclarval forms, however, are carriedthrough the traveling screens by thecooling water pump flow.Throughout the 1250 to 1600 footlong, 12 foot by 12 foot squareconcrete conduits over 50,000

square feet of settlement surfacearea is available for larvae of

e of predators and the downstream heatfrom the walls,

ocean, comparable growth would requireyears to reach the same size. Pollicipespolymerus, a gooseneck barnacle formsdense "shag rug" communities on thewalls, as welf. The perfect environinentof rich food resources from the fast

flowing waters allow these to reach 6-8inches in half a year compared to wildforms that grow 1 inch in a couple ofyears.

Heat treatment, the recycling of heatedeffluent through an inactive coolingwater conduit, was used as the primarymacro-fouling control method from 1985

Figure 2 Acorn barnacles, Megabalanus caifornicus, to January 1989 During this period 13in clump formations found at Diablo Canyon. heat treatments were used to control

biofouling. Barnacles killed during heattreatments were not physically removed from the system due to the adhesion of empty shells to the walls.Over time recolonization by new larvae on these empty shells resulted in the forination of clumps thatweaken their attachment to the basal shells, break off, and effectively plug condenser tubes. Because of&equent de-rates to clean condensers following heat treatments the process was replaced in 1989 by inanualscraping conducted during 50'/0 power curtailments. Crews removed up to 100 tons of biofouling materialwhen inanual scraping a conduit, Both of these control strategies required isolating one of the CWPs.Without this source of cooling, the power plant was also required to ramp a unit to 50/0 power and isolatea CWP for 30 to 72 hours each. During each episode thousands of megawatts were not produced andmillions in generation revenue were lost per fuel cycle,

Strategies to reduce these losses improved in 1992 when an automated chemical injection system forcontrolling biofouling was installed at the intake. Acti-Brom Nalco Chemical Co,! is a proprietary sodiumbromide solution that blends with a surfactant and sodium hypochlorite bleach! to inhibit settlement andgrowth of biofouling organisms. The solution is injected several times daily, at a rate to maintaincompliance with NPDES discharge limits. The biocide solution has the additional benefit to plant operationof controlling micro-fouling inside the condenser tubes. Generation losses were minimized for complete fuelcycles using Acti-Brom, The exception was removal of biofouling &om areas not chemically treated, suchas portions of the intake forebays,

Concrete walls and dividers located seaward of the chemical injection point could not be treated by Acti-Brom. Moving the injection point was cost prohibitive and of dubious effectiveness. Additional controlinethods were required to stem the settlement and growth of barnacles in the forebays and on stationaryportions of the steel traveling screen &ames. Foul-release coatings were investigated as a passive contro1strategy for these remote, continuously submerged locations,

Other Power Plants

To deinonstrate the effectiveness of foul-release coatings against freshwater biofouling, I contacted twornid-west power plants; Arkansas Nuclear One, on the Arkansas River west of Russellville; and DuaneArnold Energy Center DAEC! on the Cedar River near Cedar, iowa. Both plants use silicone coatings asproactive measures to inhibit Zebra Mussel colonization on pumps, trash bars, traveling screen frames, andintake pits.

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FOUL-RELEASE COATING APPLICATIONS

For millennia coated surfaces have been used to deter biofouling from the hulls of marine vessels andsubmerged structures. Coatings einploying toxic chemicals and inetals e.g. copper, zinc, or tin! have beensuccessful and some are stil] in wide use today. These coatings, however, are not acceptable at coastalfacilities, because of discharge restrictions. Toxic elements damage the environment and lead to long termburdening of sediments, and tissues of animals and plants. Research and development of non-toxic forms ofbiofouling control produced nove t materials that limit the attachment and growth of fouling organisms. TheDiablo Canyon Biology Laboratory has participated in coating research and testing since 1986. Many typesof coatings were tested, but few satisfied the discharge restrictions while also deterring biofouling. Thehydrophobic and hydrophilic slick surfaces of silicone elastomeres proved to be the most successfulproducts tested in the lab.

m>calmi ection po>nt

Biofouling control methods at Diablo focus on specific sites in the cooling water system and are dependentupon the operational status of the power plant. Coated test plates successfully inhibited settlement ofbarnacles Sommerville and Steinert 1990! and gave promise of their eA'ectiveness in larger applications. In1989, a pilot program began in a Unit 1 cooling water conduit. A paired comparison of two coatingproducts Kansai Biox and Chugoku Bioclean! was assessed using 360 square foot patches on the walls

and ceiling within the conduit, TheDiablo Canyon Intake Structure coatings were applied directly on

sand blasted, warmed, and

dehumidified concrete. The resultantTravalnp'l hI~ Scf86h coated surface reflected the rough

exposed concrete created by blasting.Q After examining the rough texture,

PG&E coating specialistsI I recommended application of surface

fillers to smooth the surface and fill-

I tp Power in imperfections "bug holes" !exposed by blasting. In 1990, asecond trial comparing thtee silicone

F~ � foul-release coatings Kansai Biox,Wee Chugoku Bioclean, and International

Intersleek! began in a Unit 2 conduit,The concrete surface was sand-

blasted to remove barnacle base-Figure 3 Cross section d'agrain of cooling water intake plates and provide the appropriatestructure depictutg screening and Pumping eQuiPment. Section rofile for coatin lesion Thecoated with Biex shown in grayed area. rough surface, in these six test

patches, was first covered with a wet-tolerant epoxy surfacer. The foul-release systems, comprising primer and silicone coat s! were sprayedover this first layer. The resultant surface was smoother with few "bug-holes" compared to Unit 1applications. Over the elapsed 8 or 9 years since application, observations within the dewatered conduitsreveal that two of the coating products work well over the initial few years. After long exposure the filledand smoothed surfaces in Unit 2, however, remained completely See of barnacles and mussels for the entire8 years exposure and continue to perform well. Although Biox and Bioclean performed well in these sideby side comparisons, Biox was selected over Bioclean. Several 3 to 4 inch diameter blisters developed inthe Bioclean patches while none were observed in any Biox patches. The third coating tested, Intersleek,fouled lightly in the first year and performance declined in following years.

Based on this experience, in 1996 and 1997 PGPE funded a project to coat portions of the intake forebayswith foul-release silicone to augment the DCPP biofouling control program. To minimize effort and cost,yet produce an efficient control measure, the areas to be coated in the forebays were selected based onfouling community distribution and growth patterns. Over many years, I noted the lower ] 0 to 15 feet ofthe forebay fouled with repeated severity with large Megabalanus, Pollicipes, and Mytilus edulis baymussel!, This zone corresponds to the area of greatest flow past the traveling screens and into the pump

bowl Figures 3 and 4!. Areas outside thePlan View Intake Structure traveling screen were precluded because

fouling is light and any detached shells arecaught and removed by the rotating screens.

Coo P The floor of the forebay and pump bowl werenot coated either because of slipping hazards.Based on design drawings the areas to becoated encompassed approximately 2,500 sfin each forebay.Foui-Rel

In the current configuration, this strategytargets chemicals in the main conduit�coatings in the forebays and some smallpipes, plus localized heat treatment in smallpipes. The an-line control methods arecomplimented by manual scraping during the30 to 40 day refueling outages between fuel-cycles. The program has proven successfulover 18 to 21 month fuel-cycles.

Tl'BYelP

Intake Cove

Figure 4 P!an view diagram of coohng waterintake structure depicting wall surfaces coated byBiox.

Surface Preparation

At the floor level, located approximately 35 feet below sea level, the intake forebay environment is wet,humid and difficult to access. Excessive moisture and humidity challenge the coating specialist to find ways

to apply, cure, and adhere coating to a surfacethat has been exposed to seawater for over 20years. In preparation to apply a coating systemin the intake, several trial applications in wetconditions were needed to develop theappropriate system that would functionsuccessfully. From these series of tests, a wettolerant epoxy system was chosen to fill in themajority of imperfections created by blastingthe concrete and bond with the over-layingfoul-release systems. The surface to be cleanedwas very rough with the residual base platesleft over &om scraping by divers Figure 5!.

Figure 5 Rough shell base plates ou concrete wallprior to hydro-blasting portions of the intakeforebay. Pictured is the four inch diameter PVCchemical injection pipe.

85

Following the divers, gates stop-logs! were lowered and the intake forebays dewatered by pumping. In theconfined space maintenance crews scraped and removed the remaining macrofouling and erected

scaffolding to assist the paint crews. High pressure water blast with injected sand wasused to remove barnacle bases and provide anadequate � mil! profile for coating adhesion. Alonethe 5,000 to 6,000 psi water pressure wasinadequate to remove the tenacious base plateswithout injecting Monterey sand by venturi. Theaddition of sand allowed the cleaning and profilingto proceed rapidly. The blast operator could notblast one spot for too long, Otherwise, the concretewould erode and create deep depressions which aredifficult to rebuild without extensive labor. As

described above, blasted concrete reveals many

Figure 6 Hydro blasted concrete waII showing small imPerfections Figure 6!. Following theexpose ug- o es, pits, an inipe ections. blasting all water was pumped from the intake and

all sand was removed from the scaff'olding, wallsand floor. Cleanliness is imperative so that debris does not contaminate the finished silicone surface.

Additional water control, due to leaking gates, required a series of dams be set up within the travelingscreen bays. This accumulation of water was automatically pumped between adjacent bays and then outthrough the single surface access hatch. Temperature was raised and humidity decreased by supplyingwarm 95'F! diy air from topside dehumidifiers �50 cfin!. This raised concrete surface temperatures from55'F up to 60-70'F and dropped relative humidity from100% to levels between 70 to 50%,

Coating System Application

After cleaning the concrete, establishing properenvironment and ventilation, a thick, wet-tolerant epoxy Bio-Dur 561, Thin-Films Inc.! was troweled on to thedried concrete. Approximately 70 kits of Bio-Dur 561were applied over the 2,500 sf in each forebay. In eachforebay the troweling process required about three shiftsto complete and 10-15 hours to cure Figure 7!. After thecure, a second wet-tolerant epoxy Bio-Gard 251, Thin-Films, Inc.! was either rolled or brushed on to the walls,and valves Figure 8!. Both of these 100% epoxyproducts have similar formulations, except for differingamounts of filler fibers. Within a work shiA the Bio-Gard

was sufficiently cured to begin application of the foul-release products.

The Biox silicone system is composed of three coatinglayers. The first layer sprayed-on is Biox Primer. It isgray and provides a chemical bond between the baseepoxies and next silicone layers. The primer was appliedat 2-3 mils wet. The primer and epoxies were allowed tocure for 24-hours to ensure release of volatile

Figure 7 Coating applicator troweling Bio-Dur 561 on a concrete wall in DCPP intake.

components VOCs! in the primer and complete chemicalreactions in the epoxies. Dehumidification and ventilationwere maintained during the curing interval.

Two silicone layers were sprayed over the primer Figure9!. Both are clear coatings that require detailed attentionto assure complete coverage. The first layer is Biox 1and is applied 4 to 7 mils wet. Aller a 5 to 6 hour cure,the second, Biox 2, coating is sprayed on at 8 to 11 milswet. Several personnel are required to support thecoating applicator when applying these clear coatings.Despite proper application of the lower four layers, if thelast Biox 2 coating has any gaps those missed spots willreadily foul. This demonstrates the active measuresengineered in the coating. Besides the slick siliconesurface, Biax 2 coating also has an "oozing agent" thatfurther protects the surface from settling larvae. If ananimal does adhere, as it grows adhesion becomes moredificult to maintain. Water flow eventually removes thegrowing pest, To prolong the effectiveness of the coatingsystem the maximum Biox 2 milage recommended by themanufacturer was applied by the painters.

ln these applications, conducted in a deep confi~edspace, safety was of paramount concern. All the Biox Fi re 8 A lication of Bio-Gard 25l e ox

coatings contain various amounts of volatile compounds on a service water cross-tie valve and intake

such as toluene, xylene, and gasoline, To prevent the walL

build up of these potentially explosive vapors a large exhaust fan drew fresh air &om the intake top deckthrough the forebays. The contaminated air was discharged out toward the ocean, away froin concurrentwork. Within the forebay, lower explosive detection LDL! monitors were worn by the applicators, supportcrew arid top side personnel. Rarely, did the LDL monitors alarm while applying any of the coating layers.

On the few occasions of alarm, a shortcessation �0 sec! of spraying was all thatwas required to clear the air. All personnelwore full-faced respirators fitted withchemical and pesticide grade cartridges.

In San Luis Obispo county, Air PollutionControl District APCD! regulators limit thevolatile organic compounds VOC! releasedto the atmosphere &om coatings. Theregulated maximum is 3.51 pounds VOC pergallon of coating. All epoxy and Biox coatingused in this application complied with thisregulation except Biox 1. VOCs in Biox I are3.63 pounds per gallon. San Luis ObispoAPCD Rule 4.0.2 excepted the excess, butlimited application to not exceed 8 pounds

VOC per hour or more than 40 pounds VOC per day. Based on these limits Biox 1 was applied over twodays in hour ly shifts. As a note Kansai has developed a new formulation, Biox L which meets the VOC

limits for San Luis Obispo County. Biox L can also be applied directly over any two-part epoxy. Thisimproves coating time, labor, and material costs. Techmcal specifications at DCPP, however, were writtenfor the earlier formulation of Biox 1 and Biox 2. Initial testing of Biox L is promising, but extensive long-term data prevented changing products when funding and planning were available.

EVALUATION OF FOUL-RELEASE COATING

Coating Performance

Coating systems were applied to Unit 2 forebay walls in May- June 1996. Unit 1 forebays were coated inFebruary-March 1997. Due to the success of the biofouling control program, access to the Unit 2 forebaywas not available until the following refueling outage inFebruary 1998. After 630 days exposure, the coating wasinspected and found in good condition Figure 10!. Attachedbiofouling was limited to thin patches of encrusting diatoms,hydroids and bryozans. These are thin and do not representpotential hazard to the condenser if detached. Isolated largebarnacles were also observed scattered over the coated surface.

These one hundred or so barnacles gained a foothold as larvaeby attaching in small depressions that were not completely filledby the Bio-Dur. Compared to previous inspections of untreatedsurfaces the coated forebay walls were essentially clean of anybiofouling.

At the same time Biox coated surfaces within the travelingscreens and small diameter piping were examined. Travelingscreen frames, exposed for 1 to 2 years were lightly fouled, butgenerally clean of biofouling. Flow rates through the screens,however, are slower than through the forebay and conduits. TheU-beams also provide places for barnacles to hide Rom theflow. Despite a higher rate of fouling on these beams comparedto flat walls, the coating success was obvious. A Biox coatedsection of ten-inch diameter pipe supplying cooling water to theintake coolers was also free of fouling after 21 months ofservice.

Figure 10 Biofouling on Biox coatedwalls in Unit 2 intake torebay February1998 after 630 days exposure to flowingseawater.

Program Evaluation

Ss

A reduction in power generation curtailrnents de-rates! motivates improvements in biofouling controlmethods at Diablo Canyon. The success of these programs can be judged by comparing generation records.Since 1985, each biofouling contra! event has been documented based on cause and control method. AtDCPP cause is distiriguisked between internal cooling water debris i.e. barnacle shells that have grown anddetached from the walls! and external debris such as kelp and seaweed that pass by the screening at theintake. Figure 11 compares the average daily loss in generation MWe! for each fuel cycle. A fuel cycle isthe interval between core refueling outages. The daily loss attributed to controlling "All Biofouling" whichincludes internal and external sources fluctuates over time because of random oceanographic eventsassociated with the productivity of seaweeds and timing of high swells. The control of shell debris withinthe conduits is more manageable &orn a biofouling control view. The first four fuel cycles losses seemvariable in success, Not shown, however, are two major cleaning events conducted when the plant was off-

line for mechanical repairs. If those events were included cycle 3 averages would exceed 400 MWe perday. Beginning in cycle 4 changes in strategy begin to reflect

improvements in controlling shell debris by manually cleaning the conduits during weekend curtailments at50'lo power. Since cycle 6, in 1992, control measures have caused the average losses to approach 170MWe per day. This alone is a benefit to the overall gross generation that typically produces 27,400 MWeeach day.

The recent successes in reducing generation losses are accompanied by longer operation during each fuelcycle Figure 11!. During the latest fuel cycle this program realized an equivalent control success comparedto the previous two despite extending the fuel burn by 130 days. This is attributed to coating the intakeforebays and traveling screen &ames with Biox silicone, plus the existing chemical and manual cleaningmethods,

Improvements in generation reliabi/ity will help Diablo to become more competitive in today's environment

~ All 8iofouling Shell Debris Only ~ Fuel Cycle d!

~ 700700

I sooIsoo >

600

4 5

FUEL CYCLE NUMBER

Figure 11 Average daHy loss in generation attributed to biofouling control measures compared tothe duration of fuel cycles. Shown are biofouling related curtailments that encompass combinedmeasures to control all internal barnacle shells! and external seaweed! debris and losses attributedto de-rates caused by shel! debris alone. Fuel cycles operate between reactor core refueling outages.Two major shell debris related losses occurred during forced outages FO! in cycle 3 and one lncycle 6.

of utility deregulation. As changes in operation strategy occur, a variety ofbiofouling contro1 strategiesneed to be available. Using non-toxic silicone coatings has proven effective and cost efficient for portionsof the cooling water system, At this time coating controls measures are blended with several other strategiesthat include chemical injection, manual scraping, and localized thermal treatment. The variety of successfulcoating applications at Diablo demonstrates the flexibility and long term availability this biofouling controlstrategy offers to coastal generating facilities.

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500

O 400

X O 300I-

200

100

400 Lr

0300

200

1-0

Other Facilities

Success at a coastal utility utilizing silicone coatings ismatched at two freshwater facilities. Even though zebramussels have not invaded the Cedar River in Iowa, coatings atDuane Arnold are intact on trash racks, pump pit walls andpumps. Zebra mussels have invaded the cooling watersutilized by Arkansas Nuclear One. Figure 12 shows theeffectiveness of a silicone coating that prevents the attachmentof Dreissena po ymorpha on coated walls and pumps.

REFERENCES

Sommerville, D.C. and F.L, Steinert. l990. Development ofalternative macrofouling control methods for Diablo CanyonPower Plant. EPRI International Macrofouling Symposium,

December 4-6, 1990 Orlando, Florida. Figure 12 Pumps at Arkansas NuclearOne. Lower, coated with a silicone

coating, is clean after one year. Upperpump, coated with epoxy, is covered byzebra mnssels.

90

MUNICIPAL POTABLE WATER IIOLLUSCICIDE

FOR THE 21 CENTURY

Jim Dicksa

Calgon CorporationP.O. Box 1346

Pittsburgh, PA 15230 U.S.A.

ABSTRACT

In the fourth quarter of 1995, Calgon VeliGONr" products received registration from theEPA as an approved molluscicide for use in controlling zebra mussels in potable systemraw water intake lines. The VeliGON products are varying molecular weights ofdimethyl-diallyl-ammonium chloride DMDAAC! which have also been certified by NSFfor the use in potable drinking water.

This paper discusses the use of non-oxidizing molluscicide VeliGON TL-M at the NorthShore, WI and Menominee, Ml Water Treatment Plants in 1997 and early 1998 Thesefacilities draw their raw water &om western Lake Michigan. Efficiency data in regard tozebra mussel kill will be presented. Presented operational data utilizing particle countersand streaming current detectors will i~elude inorganic chemical feed reduction, the impactof increased filter runs as well as reduced sludge generation.

THE "MECHANICAL" FILTRATION OPTION

FOR ZEBRA MUSSEL ELIIIINATION

Steve SpringerAmiad Water System Technologies

P.O. Box 5547

Oxnard, CA 93031-5547 U.S.A.

ABSTRACT

The control of zebra mussels and other mollusks with mechanical filtration has 1ong beendebated as a systematic and efficient means to provide a 100'/0 barrier for all "viable" lifeforms of this, and other "aquacritters". Now, however, based orr independent testing andactual filter system installations, this mechanical alternative for preventing their intrusioninto piping and water systems has been proven to be reliable, economical and ecologicalfriendly. Further, it is now substantiated and documented that a specially-fabricated weaveof 40-micron "absolute" stainless steel screen will stop all viable life forms of mollusks.However, the success of stopping the mollusk provides only half the process, Once theseorganisms have been collected in this 40-micron weave, they must be removed anddischarged as easily as they were collected. The most significant advantage to themechanical filtration option is ecological, With mechanical filtration, there are NOchemicals or "special" handling of the wastewater, Without the use of chemicals the waterdoes not have to be "de-chemicalized" or sent to sewer. With mechanical filtration, both

the "waste" water and the system water can be discharged or used without any potentialimpact on the environment or their ecosystem s!.

The other significant advantage is that of cost Although mechanical filtration can be asignificant up-front capital expense, the effect on future annual maintenance budgets is veryminimal. Unlike that of chemicals that require ongoing expenditures in purchasing costs, storagecosts, and elimination costs, "screen" mechanical filtration has no medium to replace each yearand very little arrnual service maintenance.

For reference purposes, a 5000 GPM system, requiring 40-micron absolute filtration, willhave an average up-front cost of about $40 to $45 per gallon of filtered water. This ofcourse is very subjective due to the variance in factors not addressed by filtration alone;i.e., control systems, screen micron selection, variations in water quality, and mostimportantly system flow rate. Although 40-micron absolute is necessary for 1000loremoval of all life forms, 100/0 removal may not be a requirement for every system andthis choice can have a significant impact on cost.!

For over 30 years, Amiad Filtration Systems has been providing innovative and reliablefiltration systems. During that time Arniad was able to develop and perfect the hydraulicand mechanical symbiotic! relationship between screen filtration and screen cleanirigtechnology. Over the past three years, Amiad has proven that their "technology" infiltration systems does in fact provide the mechanical protection required to prevent zebramussel intrusion into waterways and piping systems. Amiad's patented automated self-cleaning screen filters provide both the mechanical filtration requirement of 40-micronabsolute and a "scanning" cleaning method that efficiently and electively cleans 100'/a ofthe screen surface.

92

independent testing done by Acres International for the State of Michigan substantiatedthis in 1993. These tests demonstrated not only the integrity of the screen but, the screencleaning systein. A filter system three filters with a combined filtration area of 4560sq/in.! was then installed at the Mud Creek Irrigation District for the control of zebramussels for irrigation water. The filter system was designed to handle a flow rate ofapproximately 7000 GPM and provide irrigation water to a ditch system being pumpedfrom Saginaw Bay � Lake Huron. The system saw its first successful "full" season ofoperation in 1997 and has been monitored under the guidelines provided by Acresinternational and the State of Michigan,

Further, in 1996 Amiad provided an even more unique zebra mussel filtration system tothe State of Vermont for the State fish hatchery. This system was unique in that it not onlyrequired 40-micron absolute protection, it had to provide protection from two independentgravity water sources through the same filtration system being supplied &om low pressuresupply lines, This required a very sophisticated control and pumping system that interfacedwith the filters to supply "mollusk free" water to the hatchery, This system just finished itsfirst full season of successful operation.

Mechanical filtration is the effective option to stop zebra mussels!

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EFFECTS OF AN ORGANIC ANTIFOULING COMPOUND MEXEL 432!ON BOTH SETTLEMENT OF Dreissena LARVAE AND SEVERAL

BENTHIC INVERTEBRATE SPECIES AND ON GROWTH OF ADULTMUSSELS � FIELD AND LABORATORY EXPERIMENTS

Nathalie Czembor, Laure Giarnberini, Jean-Claude PihanEcological Research Center, Universite de Metz

1, Rue des Recollets, B.P. 411657040 Metz, Cedex 01, France

ABSTRACT

We report in this presentation, both field and experimental studies realized in order toevaluate the molluscicidal and antifoulirig properties of an organic compound, Mexel 432 Mexel Society, France!, financially supported by Electricite de France,

A field study has been conducted for three years in the Moselle River Metz, northeast ofFrance! to test the efficiency of Mexel 432 in inhibiting settlement of several species ofmicro-invertibrates. The tests were developed in a flow-through system with water directlypumped into the river. Treated tanks received three intermittent injections �5 min.! of 4mg/L of product daily. The results showed a high reduction of the settlement of Dreisseriaveligers and other organisms Protozoa, Hydrozoa, Rotifera, Chiromidae! in the bioboxes,

On the other hand, the growth of exposed adult zebra mussels dropped to 75'/0 both in fieldand laboratory tests intermittent injections of 0.5, 1, 2 mg/L!. The small mussels under12 mm! were the most affected. After five months under exposure conditions, the mortalityrate reached 80'/0 three injections, 15 minutes, daily!.

This study and previous results showed the Mexe! 432 is efticient in controlling andpreventing settlement of zebra mussels and reducing general fouling.

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U.S. AND INTERNATIONAL REGULATORY INITIATIVES REGARDINGBALLAST WATER AND SHIPPING

Lt. Lawrence Greene

U.S. Coast Guard

Commandant G-Mor-2!2100 2nd Street S.W., Room 2100Washington, DC 20593 U.S.A.

ABSTRACY

In 1993, the U,S. Coast Guard published final rules in the Federal Register requiring allvessels who enter the Great Lakes, lrom outside of the Exclusive Economic Zones of theU.S. and Canada, to practice ballast water management. These regulations were a result ofthe Nonindigenous Aquatic Nuisance Prevention and Control Act of 1990 NANPCA!,The National Invasive Species Act of 1996 NISA!, which reauthorized and amendedNANPCA, was signed into law in October, 1996. NISA was designed to minimizeinvasions of nonindigenous aquatic species into all U.S. Ports, The law calls for aNationwide ballast water management program for all vessels which enter U.S. waters. InDecember 1997, a Notice of Proposed Rulemaking NPRM! was published in the FederalRegister announcing the new ballast water regulation. The current status and generalprovisions of the U.S, regulations, as well as regulatory activities of the InternationalMaritime Organization IMO! will be discussed.

95

BIOLOGICAL INVASIONS AND OPPORTUNITIES FOR THEIR

REGULATION ON THE WEST COAST OF THE UNITED STATES

Andrew 1V. Cohen

San Francisco F stuary Institute! 80 Richmond Field Station, 1325 South 46th Street

Richmond, CA 94804 U.S.A.

The introduction of exotic organisms may constitute the largest current threat to biological diversity inmarine and freshwater ecosystems. Invasions are a common � m some studies, the most commoncontributing factor to the endangerment and extinction of freshwater fish, crayfish and other freshwaterorganisms Miller et al. 1989; Richter et al. !997!. Although few global extinctions have yet beendocumented in marine systems &om any cause Carlton et al. 1991; Carlton 1993!, several marineinvasions have been credited with large reductions in native organisms, potentially leading to at least localextinctions e. g, G!ude 1955; Travis 1993; Kimmerer et al 1994; Grosho!z & Riiiz 1995!. Biologicalinvasions are generally irreversible, and in both &eshwater and marine ecosystems the rate of invasions hasbeen dramatically increasing Mil!s et al. 1993; Cohen &, Carlton 1998!, and will likely further acceleratewith our current headlong rush toward promoting global markets and international trade.

On the West Coast of North America, aquatic invasions have been most intensively studied in the SanFrancisco Bay/De!ta Estuary, where the establishment of over 200 exotic species has been docuinented,including plants, protists and invertebrate and vertebrate animals Cohen & Car!ton 1995!. Another 100-200 species should be considered cryptogenic � species that, based on current knowledge, could be eithernative or exotic Car!ton 1996!, Aside &om the sheer number of exotic species, they dominate manyhabitats, accounting for 40'/0 to ! 00'/o of the common species at many sites in the Estuary. The majority ofthe organisms living on the muddy bottom of the Bay and on the sides of the docks derive from the northernAtlantic or the western Pacific. Most of the Delta's fish are native to the eastern United States Moyle1976!. The crustacean zooplankton fauna in the northern part of the Estuary is now primarily composed ofspecies &om across the Pacific, and almost every year seems to bring us yet another Asian copepod ormysid shrimp Orsi 1995!. Many of the introduced gelatinous zooplankters, on the other hand, are fiom theBlack Sea e. g. Mills & Sommer 1995!. Our common crabs include species from Europe the green crabCarcinus niaenas!, the eastern United States the mud crab Rhi thropanopeus harrisii!, and China themitten crab Eriochei r sinensis!; our common clams come &om the Western Atlantic and the WesternPacific; our mussels from the Atlantic, the Mediterranean and the South Seas; our snails &om the WesternAtlantic; and our sea slugs &om Asia and New Zealand Cohen &. Carlton 1995!. As with the humanpopulation of California, it is often ififEicult to find a native.

The introduction of these exotic organisms has had both ecological and economic impacts. Introducedcompetitors, predators or parasites have contributed to declines in various native species, The flows ofnutrients and contaminants have been dramatically a! tered A!pine & Cloern 1992; Werner & Hollibaugh1993!. Exotic plants have invaded beaches, tide flats and marshes Grossinger et a!. ! 998!, and threaten toalter key winter feeding sites for the Pacific F!yway's inigratory shorebirds.

Some intentionally introduced fish and shellfish supported commercial fisheries for a time, a!though theyonly support a recreational harvest in the Estuary now. Overall, most of the econoinic effects have beennegative, especially for unintentiona!!y introduced species. One exotic marine wood-boring organism theAtlantic shipworin Teredo navalis! caused an estimated $2-20 billion of damage in two years' time Cohen1996!. Hull fouling, due in part to introduced organisms, increases fuel consumption and vesselmaintenance costs, as has been extensively documented by military and other authorities e. g. Woods Hole

96

1952; Haderlie 1984!. California has spent millions trying to control introduced aquatic plants and fish inthe watershed. Current control activities for exotic fouling, plants and fish! release toxins into theenvironment, with additional environmental, occupational and possibly public health risks. By contributingto the endangerment of native species and by continually changing the biota and making the ecosystemmore difficult to manage, the introduction of exotic species threatens to hinder water diversions and othereconomic activities in the Estuary, with implications for the whole of California's economy Cohen 4Carlton 1995!.

Exotic species have been reported in virtually all harbors and bays along the Pacific Coast Carlton 1979!.Once established in one bay, organisms may readily invade another. For example, the European green crabCarcinus niaenas, first reported in 1989-90 fi'om San Francisco Bay, was found in estuaries from ElkhornSlough to Humboldt Bay by 1995, reached southern Oregon in 1997, and was found in Willapa Bay,Washington in 1998 Cohen et al. 1995; Grosholz and Ruiz, 1995; Miller, 1996; N. Richmond, pers.comm.; A. Cohen, unpublished data!. Exotic species may also deploy out of the bays to invade the opencoast. A predatory New Zealand sea slug Philine auriformis that was collected in San Francisco Bay in1992 Gosliner 1995! and is now found from southern California to Bodega Harbor, has become the mostcommonly collected sea slug in the Southern California Bight D. Cadien, pers. comm.!.

Issues of this sort have resulted in some efforts to deal with the more egregious mechanisms that areimporting exotic organisms into U. S. coastal waters, particularly the transport of organisms in ships'ballast water, ln the early 1990s, ships entering the Great Lakes and upper Hudson River from overseasports were required to adopt measures, such as the exchange of ballast water on the high seas, to preventthe discharge of exotic species. In 1996, oil tankers selling Alaskan oil overseas became subject to similarrequirements under a federal rule. However, despite those promising initial steps, the disappointingNational Invasive Species Act passed in 1996 only made it "officially voluntary" for ships to do anythingabout their discharge of exotic species.

Responding to that display of federal inaction, efforts have developed at state and local levels to regulatethe dumping of exotic species into coastal waters. In May 1997, the San Francisco BayKeeper, assisted bythe Environmental Law Community Clinic, petitioned California's Regional Water Quality Control Boardsto have ballast water discharges regulated as a waste discharge of a biological pollutant under California'sPorter-Cologne Water Quality Act. In February 1998, the San Francisco Bay Regional Water QualityControl Board listed exotic species discharged in ships' ballast water as a high priority pollutant causingimpairment of the water quality of San Francisco Bay, under provisions of the federal Clean Water Act.And in March 1998, the Center for Marine Conservation and seven other environmental and fishingorganizations charged that the Port of Oakland is required under state and federal environmental law tomitigate for any increase in the number of exotic organisms carried into San Francisco Bay in ships' ballastwater as a result of the Port's proposed dredging and expansion project. Several other similar actions areapparently pending in California, Oregon and Washington. Indeed, if the federal government waits muchlonger to address this issue through effective regulations, it may ultimately be determined through litigationthat shipping companies and other importers of exotic species! are financially responsible for the impactsof the exotic species that they release.

REFERENCES

Alpine, A. E. k J, E, Cloern �992! Trophic interactions and direct physical effects control phytoplanktonbiomass and productio~ in an estuary. Limnol. Oceanogr. 37�!: 946-955.

Carlton, J. T, �979! History, Biogeography, and Ecology of the Introduced Marine and EstuarineInvertebrates of the Pacific Coast of North America. Ph. D. thesis, Ecology, University of California,Davis.

97

Carlton, J. T, �993! Neoextinctions of marine invertebrates, Amer. Zool. 33: 449-509,

Carlton, J, T. �996! Biological invasions and cryptogenic species. Ecology 77: 1653-1655.

Carlton, J. T., G. J. Vermeij, D. R. Lindberg, D. A. Carlton & E. C. Dudley �991! The first historicalextinction of a marine invertebrate in an ocean basin: the demise of the eelgrass limpet Lottia alveus. Biol.Bull. 180; 72-80,

Cohen, A. N. �996! Biological Invasions in the San Francisco Estuary: A Comprehensive RegionalAnalysis. Ph. D. dissertation, Energy and Resources, University of California, Berkeley,

Cohen, A. N, & J. T. Carlton �995! Nonindigenous Aquatic Species in a United States Estuary: A CaseStudy af the Biological Invasions of the San Francisco Bay and Delta. U. S. Fish and Wildlife Service,Washington DC.

Cohen, A. N. & J. T. Carlton �998! Accelerating invasion rate in a highly invaded estuary, Science 279:555-558.

Cohen, A. N., Carlton, J. T. & Fountain, M. C. �995! Introduction, dispersal and potential impacts of thegreen crab Carcinus maenas in San Francisco Bay, California. Mar. Biol. 122, 225-238.

Glude, J. B. �955! The effects of temperature and predators on the abundance of the soft-shell clam Myaarenaria in New England. Trans. Am. Fish. Soc. 84' .13-26.

Gosl incr, T. �995! The introduction and spread of Philine auriformis Gastropoda: Opisthobranchia! fromNew Zealand to San Francisco Bay and Bodega Harbor. Mar. Biol. 122: 249-255.

Grosholz, E, D. & G. M. Ruiz �995! Spread and potential impact of the recently introduced Europeangreen crab, Carcinus maenas, in central California. Mar. Biol. 122: 239-247.

Grossinger, R., J. Alexander, A. N. Cohen & J. T, Collins �998! Introduced Marsh Plants in the SanFrancisco Estuary: Regional Distribution and Priorities for Control. San Francisco Estuary Institute,Richmond CA.

Haderlie, E. C. �984! A brief overview of the effects of macrofouling. Pages 163-166 in: Costlow, J. D.& R. C, Tipper eds.!, Marine Biodeterioration. An Interdisciplinary Study Proc. Symp. Mar.Biodeterioration, 20-23 April 1981! Naval Institute Press, Annapolis MD.

Kimmerer, W. J., E. Gartside & J. J. Orsi �994! Predation by an introduced clain as the likely cause ofsubstantial declines in zooplankton of San Francisco Bay. Mar. Ecol. Prog. Ser. 113: 81-93.

Miller, R. R,, J. D. Williams & J. E. Williams �989! Extinctions of North American fishes during the pastcentury. Fisheries 14�!: 22-38.

Miller, T. W. �996!, First record of the green crab Carcinus maenas, in Humboldt Bay, California. Calif.Fish Game 82�!, 93-96.

Mills, C. E. & F. Sommer �995! Invertebrate introductions in marine habitats: two species ofhydromedusae Cnidaria! native to the Black Sea, Maeotias inexspectata and Blackfordia virginica, invadeSan Francisco Bay, Mar. Biol. 122: 279-288.

Mills, E. L., Leach, J. H., Carlton, J. T. and C. L. Secor �993! Exotic species in the Great Lakes: ahistory of biotic crises and anthropogenic introductions. J Great Lakes Res. 19�!; 1-54.

Moyle, P. B. �976! Fish introductions in California. history and iinpact on native fishes. Biol. Conserv. 9:101-118.

98

Orsi, J. J, �995! Radical changes in the estuary's zooplankton caused by introductions from ballast water.Interagency Ecological Studies Program for the Sacramento-San Joaquin Estuary Ãewsletter, Summer1995: 16-17.

Richter, B, D., D. P. Braun, M. A. Mendelson k L. L. Master �997! Threats to imperiled freshwaterfauna. Conserv. Biol. 11�!: 1081-1093.

Travis, J. �993! Invader threatens Black, Azov seas, Science 262: 1366-1367.

Werner, I. k J. T. Hollibaugh �993! Potamocorbula amurensis: comparison of clearance rates andassimilation efficiencies for phytoplankton and bacterioplankton. Limnol. Oceanogr. 38�!: 949-964.

Woods Hole Oceanographic Institute �952! Marine Fouling and its Prevention, United States Navalinstitute, Annapolis MD.

99

BALLAST WATER DELIVERY AND MANAGEMENT PATTERNS:CONSEQUENCES FOR NONINDIGENOUS SPECIES

TRANSFER AND INVASION?

Gregory M. Ruiz, Anson H. Hines, Linda D, McCann, L. Scott Godwin, George Smith, Melissa Frey,Laura Rodriguez aud Kim Philips

Smithsonian Environmental Research Center

647 Contees Wharf Road, P.O. Box 28Edgewater, MD 20910 U.S.A,

L. David Smith

Northeastern University

James T. Carlton

Williams College - Mystic Seaport

ABSTRACT

The movement of ballast water by ships is currently the largest transfer mechanism fornonindigenous marine and estuarine species throughout the world, but our understandingof ballast-mediated transfer and invasion is still very incomplete. The pattern of ballastwater delivery and management is variable in space and time, A port may receive ballastwater from a variety of sources regions, including water of foreign, domestic orcoastwise!, or oceanic origin. The total volume delivered, and the composition of sourceregions, vary among ports. The rate of open-ocean ballast water exchange, a recentmanagement strategy to remove nonindigenous species, may vary among vessel types andports, Importantly, these patterns of delivery and management change over time, Althoughit is clear that nonindigenous species arrive in ballast water, the differences in biotadelivered as a function of source region and ballast water management are virtuallyunexplored. Our data from ballast water arriving to Chesapeake Bay Maryland andVirginia! and Prince William Sound Alaska!, demonstrate. l! density and diversity oforganisms decrease with voyage duration, �! domestic ballast water has a higher densityor organisms than ballast water from older, foreign sources, �! domestic ballast water isan active corridor for the transfer of nonindigenous species, and �! ballast water exchangedoes reduce the density of some organisms to less than 10'!o of the original density. Thesedata begin to measure variation in biota of ballast water with different histories, but therelationship between delivery of biota and invasion success remains unclear within a singlepart or among ports. Although invasions continue to result from ballast water release, wecannot yet predict which species will become established or when establishment will occur.ln addition, it is likely that not all ports are equally susceptible to invasion from the samespecies pool, but variation in susceptibility has not been measured. A long-term goal of ourresearch program is to measure the relationship s! between supply and invasion bycomparing patterns of ballast water delivery and management to patterns of invasion bothwithin and among sites.

100

TWE ROLE QF MUD AND SEDIMENT INAQUATIC NUISANCE SPECIES TRANSFER

Philip T. JenkinsPhilip T, Jenkins k. Associates

1 Forest Hill Crescent

Fonthill, ON LOS 1E1 Canada

Aes~~cr

8allast management techniques have primarily focused on water as a vector of transfer oforganisms. This paper will describe a project that is underway to examine mud andsediment accumulation in ballast tanks, identify its potential effect on other treatmentoptions, and practical methods of elimination.

A MICROBIOLOGICAL, CHEMICAL AND PHYSICAL SURVEY OFBALLAST WATER ON SHIPS IN THE GREAT LAKES

G. Elliot WhitbyConsultant, 389 Kennedy Ave.

Toronto, ON M6P 3C5 Canada

Donald P. Lewis

Ecological Services Group

M. Shafer

Elsag Bailey Canada

Christopher J WljeyTransport Canada, Ship Safety

ABSTRACT

A total of 71 ballast water samples were collected &om 59 cargo ships in the Ports ofToronto and Hamilton and the Welland Canal during the months of November andDecember, 1995. The water samp!es were analyzed for: pH, salinity, iron, hardness, totalsuspended solids, percent UV transmittance, total and fecal coliforms, E. coli, enterococci,and heterotrophic plate count. All the ballast water samples were analyzed for thepresence of Vibrio sp. and pathogenic E. coli, Fecal coliforms and E coli were found in31 �3'/0! of the 59 ships carrying ballast water. Enterococci were present in the ballastwater of 47 vessels 80'ro!. During the survey, E. coli serotype Ol I 1, Pseudomonasaeruginosa and Provodencia rettgeri were isolated once &om separate vessels. Species ofVibrio were isolated from the ballast waters of 17 vessels. The isolates were identified as

V. fluvialis and V. alginoliticus along with three related species: Aeromonas hydrophila,3, sobria and A. caviae.

INTROG U GTION

Ballast water &om cargo vessels is considered to be a major factor in the dissemination of aquaticorganisms worldwide, The spread of the sea lamprey Petroniyzon manners! throughout the Great Lakesmarked the beginning of a long line of exotic species being introduced to &eshwaters in North American.One species that has caused enormous damage and costs is the zebra mussel Dreissenapolymorpha! Reutter,1997!. Ballast water &om marine transportation was found to be the vehicle for the transmissionof V, cholerae to Mexico from 1991 to 1992 McCarthy and Khambaty, 1994!. The potential danger tothe ecosystem, economy, and health posed by ballast water discharge prompted doinestic and internationalinitiatives for preventing ballast water-mediated invasion by aquatic nonindigenous organisms.

Following the invasion of Australia by Japanese dinoflagellates and Canada by zebra mussels, bothcountries initiated ballast water studies and began considering regulatory measures Chesapeake BayCommission Report, 1995!. In 1989 the Canadian Coast Guard set in place guidelines for the voluntaryopen ocean exchange of ballast water of ocean going vessels traveling upstream into the fresh waters of theGreat Lakes. Under these guidelines these ships could only carry ballast water taken in ocean depthsgreater than 2,000 meters with the exception of those that had not leA the North American continental shelf,The intention of the guideline was to introduce high salinity water to ballast tanks that may harbour freshwater organisms from foreign ports. In general the critical upper limit for most fresh water species is 5 to

102

8 g/L. Therefore, it was felt that the introduction of waters with a salinity of 30 g/L or greater would be aneffective control mechanism Wiley, I 997!,

In November 1990 the government of the United States released the Nonindigenous Aquatic NuisancePrevention and Control Act. In March 1991, the United States joined Canada to issue joint voluntaryguidelines. In 1990, Australia issued voluntary exchange guidelines as well.

Internationally, the United Nation's International Maritime Organization issued voluntary guidelines forpreventing the introduction of unwanted aquatic organisms and pathogens &om ship's ballast water andsediment discharge. According to these guidelines, the member states were requested but not required toexchange ballast water in open ocean areas away from at-risk coastal areas and ports. Chesapeake BayCommission Report, 1995!

The purpose of this investigation was to evaluate ballast water that could be discharged into the GreatLakes from ships of local and foreign origin fiom the standpoint of microbial safety to the ecosystem, andthe health of the human inhabitants as versus the introduction of nuisance aquatic organisms. To date verylittle attention has been given to the introduction of pathogenic microorganisms into the Great Lakes.Therefore, ballast waters were examined for human and fish microbial pathogens. Chemical and physicalparameters were measured which may have significance when considering methods of monitoring, orcontrolling microbial pathogens in ballast waters.

MATERIALS AND METHODS

Sample Collection and Analysis

The samples of ballast water were collected &om each tank through sounding pipes and access ways by thepumping action of a modified inertial pump. The samples were collected in one litre volumes in sterilebottles and refrigerated until processed within 24 hours. Twenty litres of water was passed through thesampler prior to each sample collection to prevent contamination by the previous sample.

Physical and Chemical Measurements

The following physico-chemical properties of the samples of ballast water were measured:

pH was measured using a Corning 320 pH meter and a flat surface CMS Silver Label pH Electrode &omInnovative Sensors Inc., Anaheim, CA.

Salinity was tested using a Model 140 Conductivity/Temperature/Salinity meter fiom Orion.

Total dissolved iron was measured using a Lovibond 2000 Comparator TK 100 from Tintoineter Ltd.

Hardness was measured using a Lovibond AF 424 Minikit from Tintometer, Ltd.

Total suspended solids were performed according to method 2540 D in Standard Methods for theExamination of Water and Wastewater,�995!.

UV Transmittance was measured on filtered .45'.m Gelman GN! and unfiltered water samples at awavelength of 254 nm with a Beckman DU Series 600 Spectrophotometer.

Microbiologica/ Examinati on

Heterotrophic plate coiint was performed by following method 9215A.2.b in Standard Methods for theExamination of Water and Wastewater �995!. This method uses membrane filtration and R2A as thegrowth medium.

Total coliforins were measured by the membrane filter procedure 9222B.2.a. in Standard Methods for theExamination of Water and Wastewater �995! using M-Endo-Agar LES from BDH.

Fecal coii forms were enumerated by the membrane filter procedure 9222D. in Standard Methods for theExainination of Water and Wastewater �995! using M-FC agar irom BBL.

E. coli were determined by the method of Ciebin er al. �995!. Membrane filters on M-FC BCIG agarplates were incubated for 22+ 2 hr at 44.5 + 0.2 'C. All blue colonies were counted.

Enterococci were counted by the membrane filter technique using method 9230C.Z.c. in Standard Methodsfor the Examination of Water and Wastewater �995!.

Enteropathogenic E. coli were differentiated from nonpathogenic E. coh' according to the following method.All shades of blue colonies which grew on membrane filters on M-FC and M-FC BCIG agar plates werestreaked on MacConkey Sorbitol agar Difco! for differentiation of non-sorbitol fermenting E. coli0157:H7 and on MacConkey agar BBL! at 37 'C for a purity cheek. One colony from each pure culturewas transferred to a TSA slant at 35 'C. The isolates were assessed for pathogenic potential by slideagglutination using E. coh' polivalent � to 8! antisera and the corresponding monovalent antisera DenkaSeiken, Unipath!. Isolates shown as pathogenic by the serological tests were further biochemicallyidentified to the species level by using the API 20E identification system bioMerieux Vitek, Inc.!. Healthand Welfare Canada confirmed any pathogenic E. coli.

Vibrio species pathogenic to humans and/or fish were isolated by the following techniques. Two sets ofmembrane filters on TCBS Difco! agar plates were respectively incubated at 35 'C for 24 hr and at 18 'Cfor 4 to 5 days. A nuinber of sucrose positive and sucrose negative colonies were then isolated on NutrientAgar with 2'/a NaC1 and attempts were made to identify them to the species level. The general tests forcharacterization of Vibrio species included: a! oxidase test bioMerieux Vitek, Inc.!; b! salt requirementand tolerance test using Nutrient Broth with 0'/0, 3'/0, 8'/o, and 10'/0 NaCI in two sets of tubes incubated at18 'C for one week and at 35 'C for 48 hr; c! growth in alkaline peptone water at pH 8.4 ~ 0.2 Atlas,1993!; and d! biochemical identification tests using the commercial identification systems API 20E and APIRapid NFT bioMerieux Vitek, Inc.!. Additional tests such as the string test and the swarming propertiesfor Vibrio were performed as recoinmended in the open literature Baumann et al., 1984, and McLaughlin,1995! for all the isolates identified by the API system as well as for species not included in the database ofthe API system, Presumptive V. cholerae isolates were tested serologically by rapid slide agglutinationwith V. choierae Bacto polyvalent antiserum Difco!. The quality control of the media was performedagainst Listonella Vibrio! anguillarum ATCC 19264. Any presumptive V. cholerae were confirmed byHealth and Welfare Canada.

All microbiological examinations performed by membrane filtration technique were based on duplicates ofat least two dilutions of the test sample, depending upon the turbidity of the water sample.

RESULTS AND DISCUSSION

Chemical and Physical Results and Discussion

The investigation of cargo ships carrying ballast water was undertaken in the interval of time betweenNovember 12 and December 12, 1995 and included inland or coastal and ocean going vessels. A total of71 water samples were collected from the multiple ballast water tanks of 59 cargo ships docked at the Portsof Toronto and Hamilton and the Welland Canal which connects Lakes Ontario and Erie. Two water

samples were not analyzed for total suspended solids while another sainple was not analyzed for any of thephysico-chemical parameters.

104

The pH values of the 70 ballast water samples tested ranged between 6.5 and 8 2 with a mean of 7.5. ThepH of water in the Great Lakes and the ocean usually ranges between 7.5 and 8.5 jackie and Kilgour,1995 and Austin, 1993!. The pH values of the ballast waters investigated in this study were close to thevalues reported for the Great Lakes and presented no stress for the organisms. Adjustment of the pH maybe required for chlorination but not for UV disinfection.

Salinity measurements were performed an 70 water samples with values ranging &om 0 to 40 g/L. Themean salinity in the ocean going ships was 14 g/L as versus the guideline of 30 g/L Wiley, 1996!. Figure1 shows that 70 percent of the samples were below this value and in fact 50 percent were less than 8 g/L atwhich level &eshwater organisms may be killed Wiley, 1996!. Samples collected &om ocean going vesselsvaried significantly in salinity between ships Figure 1! and even from tank to tank on a given ship asshown in Table 1. On several occasions the water &orn multiple tanks on the same vessel revealed salinityvalues varying &om 0 to in excess of 30 g/L.

I T ~ I I

O!

C C

20

10

.01

Figure l Distribution of salinities in the samples front the ballast water of the ships

Table l The level of salinity in the various ba]last containers of one of the ocean going vessel

Salinity g/LBallast Name

13.55ST

0.96ST

0.23ST

31.18ST

343P

33.8Sp

105

40

30

,1 1 5 10 2030 59 7050 90 95 99 99.9 99,99Percent of Samples

The iron content of the ballast water samples varied &orn zero to 600 mg/L with a mean of 17 mg/L. Sinceballast water is not disinfected continuously it would be possible to clean the quartz sleeves of a UV systembetween uses to prevent the build up of iron which absorbs UV light. If chlori~e was used as a disinfectantthen some form of pretreatment would be necessary to reduce the chlorine demand of the iron i f is notalready oxidized.

The determination of total hardness also revealed varied results among the 70 water samples tested. Thetotal hardness of the water samples varied from 20 to 11,000 mg/L with a mean of 2,300 mg/L. Siriceballast water is not disinfected continuously it would be possible to clean the quartz sleeves of an UVsystem between uses to prevent the build up of scale.

Sixty-eight ballast water samples were analyzed for their total suspended solids content and the distributionof values is shown in Figure 2. The total suspended solids ranged &om 1 to 6,024 mg/L with a mean of478 mg/L. Suspended solids can harbour and protect microbial pathogens making them very difficult tokill with chemical or physical disinfectants Bitton, 1994!. The ballast water would need some form ofpretreatment with a filter or coagulation and sedimentation to remove the suspended solids.

The percent UV transmittance was measured at a wavelength of 254 nm to determine whether UV lightwould be a suitable disinfectant. Figure 3 shows the results for the 70 samples before and aAer filtrationthrough a 0.45 pm tilter. The UV transmittance ranged &om 0.04 to 94 percent with a mean of 57 percentfor the unfiltered samples and 90 percent for the filtered samples with a range of 67 to 100 percent. Over70 percent of the samples before filtration had UV transmissions of 55 percent or better which would besuitable for disinfection with UV light since this is typical for the equipment which is used for wastewaterdisinfection Scheible, 1986!. All of the ballast waters had UV transrnissions greater than 67 percent afterfiltration. Removal of the iron and suspended solids would improve the UV transmittance and enhancedisinfection with UV light Scheible, 1986!.

10

1000

100

10

,01 .1

106

D!

E 0CO

ai

C S Ci.cA

Co

0

5 10 2030 50 7080 90 95 99 99.999.99Percent of Samples

Figure 2 Distribution of suspended solids in the samples froin the ballast water of the ships

100

10

0.1

L.iM i0.01

.01 .1

Figure 3 UV transmission at a wavelength of 254 uni in the samples of ballast water froin the shipsbefore and after filtration through a 0.45 Iim filter.

Microbiologii:ai Resu]ts ancf DiscussionThe results of the bacteriological sampling of the ballast water for total and fecal coliforms, E. coli,enterococci and heterotrophic plate count are shown in Table l.

The presence of any fecal indicator organisms in ballast water could indicate the presence of microbialpathogens. The results in Table 2 show that 48 percent of the samples were fecally contaminated. Fecalcoliforms and E. coli were found in 31 �3 %! of the 59 ships carrying ballast water and according to theresults based on the enterococci counts, 80 % of the investigated ships carried fecally contaminated ballastwater. Enterococci are considered a better indicator of fecal contamination of salt water because theysurvive longer Bitton, 1994!.

It has been recommended that the heterotrophic plate count of drinking water should be less than 100/mLor 500/mL especially if it is reclaimed water�rabow, 1990!. Since the ballast waters are of foreign originand not from a drinking water source they may contain exotic pathogens and should meet the standard fordrinking water. Forty-three of the samples had a heterotrophic plate count over 500 per mL and 57 wereover 100 per 100 mL.

107

S O C E th

gO

1 5 10 2030 50 7080 90 95 99 99.999.99Percent of Samples

Table 2 Results of the bacteriological sampling of the ballast water for total and fecal coliforms, E. coli,enterococci, and heterotrophic plate count.

Positive Percent

Samples Positive

Samples

Number of Mean

Samples Coloniesper 100mL

Standard RangeDeviation

Indicator

Organisms

Escherichia coli serotype 0111 was isolated and it is an enterohemorrhagic E coli. Pseudonronasaeruginosa was recovered from the ballast water and it is an opportunistic pathogen. Provodencia rettgeriwas also isolated &orn a separate ballast water sample and it has been implicated in nosocomial infections.

The presence and identification of Vibrio species was undertaken in this study to determine whether humanand/or fish pathogens such as V. cholerae and V, anguillarum, respectively were carried in the ballastwater, Since V. anguillarum is only found in sea water this could indicate the presence of pathogenicorganisms that have been picked up at sea and they could be pathogenic to fishes or other aquatic species inthe Great Lakes. Vr'brio spp. were isolated from the ballast waters of 17 vessels, One isolate was initiallyidentified as V. cholerae but Health and Welfare Canada confirmed it as A. sobria. The other isolates were

identified as V. fluvialis, V, alginoliticus and three related species, Aeronronas hydrophila, and A. caviae,

CONCLUSIONS

The exchange of seawater for freshwater will not prevent the spread of pathogenic organisms by ships as itis presently practiced.

The pH values of the ballast waters investigated in this study were close to the values reported for theGreat Lakes and presented no stress for the organisms carried by these waters.

The exchange of seawater for freshwater did not produce consistently high levels of salinity that would berequired to kill aquatic nuisance organisms.

The iron and hardness levels in the ballast water would require careful maintenance of UV disinfectionequipment.

Due to the levels of suspended solids some form of filtration or coagulation and sedimentation would berequired before UV irradiation or chemical disinfection.

The isolation of the enteropathogenic E. coli serovar 0111, Aeromonas sp., and Vibrio spp. shows thepathogenic potential of the ballast water discharge.

The ballast water should be tested for viruses and protozoan cysts that are much more resistant to theadverse conditions in the envirorunent than the bacterial indicators. The male specific coliphage could beused as a surrogate for the enteric viruses.

Since V. cholerae can enter a viable and non-culturable state it may be necessary to carry out further testsfor this organism Munroe and Colwell, 1995!.

108

Total Coli forms 69

Fecal Coliforms 69

E. coli 69

Enterococci 70

Heterotrophic 70Plate Count per*rnL

703

29

16

51

3500*

2300

70

43

94

10,600*

0 � 17,6000-400

0 -240

0-480

2 - 86,000"

45

33

31

52

65

48

45

74

REFERENCES

Atlas, R. M. �993!. Microbiological media. CRC Press Inc.

Standard Methods for the E'xamination of Water and Wastwater �995! 19 ed., American Public HealthAssociation/American Water Works Association/Water Environment Federation, Washingto~, DC, USA.

Austin, B.�993!. Marine Microbiology. Cambridge University Press, Cambridge UK.

Bitton, G, �994!, Wastewater Microbiology, Wiley-Liss, New York, USA, pp, 116 and 363.

Baumann P., Furniss A.L., Lee J.V. �984!. Genus I. Vibrio. In: Bergey's Manual of SystematicBacteriology. William k Wilkins, Baltimore, USA,

Chesapeake Bay Commission. �995! . http: www. nfrcg. gov/nas/ballast. htm. pp. 1-33.

Grabow, W,O.K. �990! Microbiology of drinking water treatment.' Reclaimed wastewater. In: DrinkingWater Microbiology, G.A. McFeters Ed!., Springer-Verlag, New York, USA, pp, 185-203.

Ciebin, B, W., Brodsky, M. H,, Eddington, R., Horsnell, G., Choney, A., Palmateer, G., Ley, A., Joshi, R.,and Shears, G. �995!. Comparative evaluation of modified m-FC and rn-TEC media for membrane filterenumeration of Escherichia coli in water. Appl. Environ. Microbiol., 61 l 1!, 3940-3942.

Mackie, G.L., atid Kilgour B.W. �995!. Efficacy and role of alum in removal of zebra mussel veligerlarvae from raw water supplies, Water Research 29 �!, 731-744.

McCarthy, S., and Khambaty, F. M. �994!. International dissemination of epidemic Vibrio cholerae bycargo ship ballast and other nonpotable waters. Appl, Environ. Microbiol., 60 �!, 2597-2601.

McLaughlin, J.C. �995!, Vibrio In: Manual of Clinical Microbiology, P.R. Murray, E.J. Baron, M.A,Pfaller, F.C. Tenover, R.H. Yolken, Ed! 6 edition ASM Press, Washington, DC, USA.

Munro, P.M., and Colwell, R,R. �995!. Fate of Vibrio cholerae Ol in seawater microcosms. Wat. Res,,30 �!,47-50.

Reutter,J.M. �997!. Importance of the nonmdigenous species/aquatic nuisance species issue. In: ZebraMussels and Aquatic Nuisance Species, F.M. D'Itri Ed!., Ann Arbor Press, lnc.,Chelsea, Michigan, USA,pp. 65-68.

Schieble,O.K. �986!. Ultraviolet radiation, In: Design Manual Municipal Wastewater Disinfection, E.L.Stover, C.N. Hass, K,L, Rackness and O.K. Scheible Ed!, U.S. Fnvironmental Protection AgencyPublication No. EPA -600/2-81-152. Municipal Environmental Research Laboratory Cincinnati, USA, pp.157-247.

Wiley, C.J. �997!. Aquatic nuisance species: Nature, transport, and regulation. In: Zebra Mussels andAquatic Nuisance Species, F.M. D'Itri Ed!., Ann Arbor Press, Inc., Chelsea, Michigan, USA, pp. 55-63.

109

TREATMENT OPTIONS FOR VESSELS NOT IN BALLAST

Christopher J. WileyTransport Canada/Department of Fisheries and Oceans

20] N. Front St., Suite 703Sarnia, ON N7T 8B1 Canada

ABSTRACT

The majority of vessels entering the Great Lakes from foreign ports carry cargo inbound toa Great Lakes port, discharge, and load at another Great Lakes port. Because these vesselsare not "in Ballast", they are not subject to either the Canadian Guidelines or U.S.regulations regarding ballast exchange. Studies have shown that these vessels still harbourlive organisms in the unpumpable slop that is left in the unballasted tanks. The totalamount of water remainirig is significantly less than that in the fully "ballasted" condition.As a result, technologies and treatments that might be unfeasable or uneconomic if appliedto a full tank may in fact be advantageously utilized for these smaller amounts.Various technologies will be discussed including a number of physical and chemicaloptions.

ANALYSES OF INVERTEBRATE FAUNA IN BALLAST WATERCOLLECTED IN SHIPS ARRIVING AT BRITISH COLUMBIA PORTS,

ESPECIALLY THOSE FROM THE WESTERN NORTH PACIFIC

C.D. Levings and G.E. PierceyFisheries and Oceans, Science Branch

West Vancouver Laboratoryc/o Pacific Fnvironmental Science Centre

2645 Dollarton HighwayNorth Vancouver, BC V7H 1V2 Canada

M, Galbraith

Institute of Ocean Sciences

G.S. Jamieson

Fisheries and Oceans, Science BranchPacific Biological Station

ABSTRACT

Between December 1995 and January 1997 we conducted the first ballast water samplingpro~m &om ships in British Columbia BC! ports, with emphasis on Vancouver. Theproject focused on invertebrates and on vessels arriving from northeast Asian ports aswell as some from the northeast Pacific.

Five of the 67 samples examined did not contain any organisms. In the others, organismsranging &om springtails to decapod zoea to medusae were obtained, with maximumabundance about 12932 animals m '. Larval forms of crustaceans, polychaetes, molluscsand other taxa were common in the samples, Cyclopoid and calanoid copepods were themost abundant taxa, accounting for approximately 50'/0 of the 35000 animals observed.One of the calanoids found Pseudodiaptomus marinus! is indigenous to Asia and hascolonized coastal ernbayments in California. Results confirmed that ballast waterdisposed of in BC waters has the potential to introduce non-indigenous species fromother parts of the North Pacific Ocean.

INTRODUCTION

In this paper we present the first results of ballast water investigations from vessels entering the waters ofthe west coast of Canada. Between December 1995 and January 1997 we sampled ballast water fromships entering British Columbia BC! ports, with emphasis on vessels arriving from the northwest Pacific.Analyses of deep sea ship arrival data, provided by the Canadian Coast Guard, showed over 1500 arrivals in1995, with the majority giving Japan, Korea, and China as the last port of call. We focused our collectionsfrom ships arriving from the northwest Pacific, since several non-indigenous species &om Asian waters havealready colonized marine and estuarine habitats on the west coast of Canada, as explained below. We alsosampled a few ships arriving &om ports south of British Columbia, specifically from Oregon, Californiaand Mexico. According to Gauthier and Steel �996!, 33.5 million m' of ballast water was dischargedfrom vessels using the port of Vancouver, accounting for 64/0 of the ballast water discharged in Canadianwaters. We emphasized sampling in ships from Vancouver Harbour, but several other ports in BritishColumbia were also visited.

Numerous invertebrate and plant species have been transferred Rom the western North Pacific to BritishColumbia through a variety of mechanisms, including Japanese oyster Crassostrea gigas! shipments, and

intentional introductions. There has been considerable effort in several parts of the world to investigatethe role of ballast water from ships as a mechanism for transporting aquatic organisms between coasts e.g. Chapman, 1988, Carlton, 1985, 1992; Kelly, 1993; Riuzetal 1997!, but there are no studiesavailable from British Columbia, Several authors have identified the ecological risk, in our particularregion, of introductions of non-indigenous crustacea e.g. the green crab Careinus maenas! Jamieson etal 1998!; copepods Cordell and Morrison, 1996! and molluscs e.g. the varnish clam Nutria liaobscurara! Harbo, 1997!, For this reason we focused our sampling on invertebrate organisms.

METHODS

Considerable effort was expended in the initial part of the project on developing arrangements forboarding vessels and sampling as this was the first ballast water prograin in our region. After boardingthe ship, an interview was also held with the Master or his representative to determine the origin of theballast water, to find out if any exchanges had occurred in mid ocean, and to get permission to sample.Our first two samples were obtained by pulling a plankton net �4 micron mesh! through the tanks butsubsequently we found that pumping water through the standpipes of the vessel was the best method toobtain water samples. The pump technique was usually acceptable to the Masters of the ships as it did notcompromise shipboard routine or ship safety. In the first part of the program December 1995 to April1996!, vessels arriving in Vancouver, New Westminster, Crouton, Nanaimo, and Prince Rupert Figure 1!were visited. During the remainder of the project May 1996 to January 1997! only vessels arriving inVancouver were sampled.

Using a portable electric diaphragm pump and plastic hose, usually 500 L of water were sampled from thestandpipes by filtering through 44 pm mesh net. Usually the hose was lowered about 7.5 m into thestandpipe, which was the maximum vertical distance the pump would lift water. The organisms retainedon the sieve were preserved in 5 lo formalin. In the laboratory, samples were decanted through a 37 pmscreen and the material remaining on the mesh was examined in the laboratory using stereo andcompound microscopes. Most were identified to class or order because of the diKculty in identifyinglarval stages. However adult calanoid and cyclopoid copepods were identified to the genus as severalAsian copepod species e.g. Pseudodiaptomus inopinus, P, marinus! have been reported from northeastPacific estuaries Cordell and Morrison 1996; Fleminger and Kramer, 1988!.

The salinity of a separate water sample was obtained with a re&actometer. Temperature was measuredby holding a hand held thermometer in the water being pumped from the standpipe.

RESULTS

Vessels Sampled and Origin of Ballast Wafer

Between December 1995 and January 1997 a total of 352 vessels were approached �98 at Vancouver, 9at New Westminster, 14 at Cro&on, 12 at Nanaimo, and 19 at Prince Rupert. Ballast water samples wereobtained from 63 vessels. On three ships, samples were obtained &om two or three different ballast tanksso the total nuinber of samples obtained was 67. The counts of organisins &om the plankton net sainpleswere not included in the data set but the presence or absence data &om them were. Copepod counts fromone pump sample were omitted because an error in subsampling.

Water from the other vessels boarded was not obtained for a variety of reasons: 1! they had disposed ofballast water before they had arrived in the harbour, 2! they had totally exchanged ballast water in mid-ocean, 3! ballast water was too low in tanks for pumping, 4! other reasons.

Twenty-seven �7! of the samples were of ballast water from Japanese ports, with the remainder fromports in Korea �!, China �!, and some ports in North America on the northeast Pacific �1!, Table 1!.Additional water samples were of mixed origin because coastal ballast water had been partiallydischarged in mid-ocean and replaced with oceanic water from the mid Pacific. For the purposes of

analyses, the samples were categorized by region of general origin, as follows Table 1!:

1. Mid-Pacific, mixed: ballast water taken on in a port in Asia, Hawaii, or the southwest coast of NorthAmerica arid then mixed with water from offshore during a Pacific crossing; includes two samples&om mid-Pacific �6 samples by pump!.

2. Northeast Asia - ballast water taken on irr a port in Japan, Korea, Taiwan, China, or the adjacent coast�8 samples by pump, 2 with net!.

3. Northeast Pacific - ballast water taken on in a port in the continental USA or Mexico �1 samples bypump!.

Sa inity and Temperature of Ballast Water

Salinity of the ballast water samples from the various areas of origin varied, with the mid Pacific samplesshowing the highest values mean 34,3 psu practical salinity units! se 0.34, range 32-37!. Ballastoriginating from northeast Asia ports was of lower salinity mean 32,1 psu, se 1.3, range 2-36!. Ballastwater from northeast Pacific ports was significantly lower mean 27.S psu, se 4,1, range 1-37! becausethis data set included freshwater from ballast taken on in the Columbia River.

Monthly mean temperatures of water samples from ships sampled in Vancouver ranged &omapproximately 6.3 C in January 1997 to 18.5 C August 1996 Figure 2!. Ballast water temperature variedseasonally, which indicated the ballast water was reflecting water conditions around the hull of the ship,not the temperature of the water when it was taken aboard.

Abundance of invertebrates Found and Relationships With Ballast Water Origin

The original data from the sampling, excluding the name of the vessel from which the ballast water wasobtained, are given elsewhere Piercey et al, 1998!.

Taxonomic composition and abundance; No organisms were found in five samples, Two of these were&orn vessels that contained ballast water &om Japanese ports, two from Korean ports, and the fifth wasfrom water taken on at a Japanese port but mixed with mid-Pacific water.

Taxa from almost all invertebrate groups were obtained in the other samples Table 2!. Calanoidcopepods �5.1'/0!, cyclopoid copepods �4.5'/a!, crustacean nauplii �0.5'/0!, cladocerans �6.0'/0!, andharpacticoirl copepods �.7'/0! were the most abundant organisms Table 2!. The number of harpacticoidsobtained is a minimum estimate because some samples were damaged during shipment of the samples toa specialist's laboratory. A wide variety of other taxa from almost all taxa were found, includingorganisms with terrestrial affinities such as springtails Collernbola!, other insects Thysanoptera andChaboridae!, and mites Acarina! Table 2!. Larval stages of polychaetes, bivalves, crustaceans, anddecapods were also observed. Diatoms, dinoflagellates, and kagments of plant material were also notedbut were not enumerated.

Samples &om the four ships sampled at New Westminster contained the highest number of animals mean3243 m ', standard error se! 1792!. The eight samples from Nanairno and Crofton combined! showedmean abundance of 61S rn se 204!. Samples from Vancouver �9! and Prince Rupert �! showedintermediate values for invertebrate abundance �021 m ' se 540! and 1592 m ' se 852! respectively.

Table 3 gives data on differences in presence or absence of the major taxa &om the three majorgeographic areas in the North Pacific where the ballast water origirrated. A few taxa were only found inballast water originating from particular regions. For example, medusae and trocophore larvae were onlyfound in samples from coastal waters of northeast Asia. The decapod megalopa larvae fiom ballast waterobtained in Asian ports were from cancrid crabs, but could not be identified to a lower taxonomic unit.

Seasona! differences: Results from the ballast water originating in Asian ports suggest there may be more

organisms arriving in ballast water from spring relative to other seasons. Unfortunately sample sizes aretoo small for statistical analyses, but mean number of organisms in each season was as follows; winter-445 m ', spring 1993 in ', summer 3333 m ', autumn 463 m ' Table 4!. Mean data &om rnid-Pacific mixand northeast Pacific ballast water sources could not be calculated because of'small sample sizes &omthose regions.

AnaIyses of Calanoid and Cyctopoid Copepods

152 individuals of the genus Pseudodiaptotnus were examined in detail, and of these 136 were P.marinus. In one sample females were carrying eggs. No individuals of P. inopinus were found. Therewere also 16 unidentifiable animals that could be placed in the genus Pseudodiaptomus, and all were fromballast water obtained in northeast Pacific ports.

Among the rest of the calanoid copepods, Acartia clausii was the most abundant, accounting for about35% of the individuals of this group. The majority of the individuals were from northeast Asia ports Table 5!. Paracalanus parvus was another calanoid that was most abundant in ballast water samplesfrom Japan, with almost 90% of the individuals reported in samples from that region. 343 individuals ofDiaptomus spp. were recorded, all &om samples obtained from two vessels that had taken on ballast waterfrom the Columbia River in Portland, Oregon. Four individuals of Clausocalanus parapergens wereobtained in the net lift sample obtained from a vessel that had ballasted in Los Angeles. 35 specimens ofthe calanoid Furytemora herdmani were found in a sample of ballast water &om Japan/Korea and thismay be ail underestimate because of a laboratory subsampling error. This species has been reported fromballast water taken on in Japan and sampled in Coos Bay, Oregon Cordell, pers comm!. 121 individualsof Pseudodiaptomus marinus, iiidigenous to Asia, were found in samples of ballast water from northeastAsia and mid-Pacific/mixed.

The inost abundant cyclopoid copepods obtained were Oithona brevicornis and Oithona sp, with about93% found in samples &om northeast Asia and mid-Pacific mixed ballast water Table 5!. Twospecimens of Oithona similis were found in the net sample from a vessel that had ballasted iii LosAngeles. Alinost all the Hemicyclops spp. were found in samples from northeast Asia aiid the northeastPacific, All individuals of Cyclops spp. �895! were obtained in two ships that had loaded freshwaterballast in the Columbia River, at Portland, Oregon Table 5!, One individual of Oncaea conifera wasfound in the sample from Japan and Korea that had been subsampled in error.

DISCUSSION AND RECOMNIENDATIONS FOR FUTURE WORK

Results of the surveys show that animals are being moved by ballast water between the major regions ofthe North Pacific. With due attention to ship safety, the precautionary approach should be taken, andmeasures iinplernented to totally eliminate ballast water disposal in BC waters. In this way, effects ofnon-indigenous species on local ecosystems, fish habitat, and species in aquaculture facilities can bereduced, There have been numerous introductions to British Coluinbia waters in the past e.g, Quayle1964!, perhaps by ballast water, but the transport mechanisms and pathways of introductions involved arenot known Elston, 1997!. Some of the non-indigenous species seem to be benign and in fact are thesources of local fisheries e.g. Manila clam, Venerupis philipinarium; Harbo, 1997!. Howevercontemporary concerns involve maintenance of biodiversity of the endeinic fauna. There is little doubtthat introductions of invertebrates via ballast water brought into Canada's Pacific coast will change thebiodiversity of local ecosystems, and in some instances this change might have deleterious effects. Anexample might be the introduction of a predator that indigenous species are not adapted to.

Assessment of the risk of introducing particular non-indigenous species such as larval forms of the greencrab would be improved with more detailed identifications of organisms in the ballast water samples.However this would be difficult because there are few specialists available who are familiar with larvalforms. These studies would be assisted by collaborative work with biologists on both sides of the Pacific.

This work cauld include exchange of data on morphological characteristics or biachemica1 genetics.Rearing of larval stages found in the ballast water could also be an effective technique for confirming theidentification of the animals and determining if the adult of the particular species is likely to be a concern.

Results from the study show that ballast water from estuaries in Oregon and California are present inships plying BC waters, and it is known that some of these estuaries have established populations of nan-indigenous organisms e.g. Cordell and Morrison, 1996!. The port of Vancouver recently establishedmandatory mid-ocean exchange of ballast water, but excluded water obtained north of Cape Mendocinoin northern California Vancouver Port Corporation, 1997!. This decision was made because it was feltthat north flowing currents off the northwest US would promote faunal homogeneity between that regionand Vancouver Harbour pers comm, Dr John Jordan, Environinental Officer, Vancouver PortCorporation!. Judging &om our data, however, this policy may not reduce the risk of trarisport of non-indigenous organisms between estuary ports, as some estuarine organisms may not have planktonic larvalstages which move with water inasses. For example, Cordell et al �992! found the Asian copepod P.inopinus in the Columbia River estuary, which is a location where one of the vessels we sampled loadedballast water. This species has also been reported fram ballast water samples from Coos Bay, Oregon, inships that had taken on ballast in Japan Cordell, pers comm!. Fleminger and Kramer �988! reportedbreeding populations of the Asian copepod P. nrarinus from two shallow embayments in SouthernCalifornia. These authors attributed the arrival of P. marinus in North America to aquaculture operationsrather than ballast water, although they as stated "we have no evidence for or against this hypothesis". Atany rate, P. marinus has nat been reported from BC waters and therefore could be potentially introducedinto the region via ballast water.

It should also be noted there may be faunal transfers from North American parts, including BC harbours,to the northwest Pacific, Although there have only been a few studies of ballast water in the latter region e.g, Ichikawa et al 1992; Chu et al 1997!, it is clear that animals from other parts of the world canestablish populations on Asian coasts e.g, Kim 1992!. These transfers could be via hull foulingorganisms as many vessels leaving BC ports are laden with bulk cargo such as coal and grain and usuallydo not need ballast water.

Our results indicate that significant amounts of water must be being dumped before arriving in BC portssince some ships enter harbour with only partially filled ballast tanks. This will vary with ship type - forexample bulk vessels will typically carry more ballast water into the region than container ships, since thelatter enter and leave port with cargo. The extent of deballasting in mid-ocean relative to coastal waters is notknown at this time, and this of course will vary with ship type, weather, and ship safety concerns. However itis likely that many ships entering the inside waters of southern BC deballast in Juan de Fuca Strait since thisareas is the first body of protected water that vessels enter after a trans-Pacific voyage, The Port ofVancouver has recently begun keeping records Vancouver Port Corporation, 1997! on where ships enteringthat harbour have exchanged water, and unless the ballast originated in mid-ocean, deballasting will not beallowed in the harbour, If mid-ocean exchange was not performed by a particular vessel, she will be requiredto leave the harbour and deballast on the north side of Juan de Fuca Strait, in the outgoing current. Disposalin the latter location may expose ecosystems on the southwest side of Vancouver Island to particular risk ofintroduction of non-indigenous species, but this aspect has not been investigated.

No program of this type implemented by Vancouver Port Corporation is operational at other ports in BC anddeballasting locations for the numerous smaller ports on the coast are generally not known. Moreinformatioii should therefore be obtained on these locations since some of them may be near sensitiveecological sites, aquaculture operations, or other places of special interest to stakeholders.

ACKNOWLEDGMENTS

This study was supported in part by the DFO Ocean Act Implementation Fund. Thanks are owing to Ms.Margo Elfert, Archipelago Marine Research Ltd. and Mr, Rob Waters, Castor Consultants, who

coordinated the sampling, We are also most grateful to ship Masters, shipping agencies, and portauthorities for their cooperation and assistance when obtaining samples. We are grateful to Jeff Cordell,University of Washington, Seattle for comments on the paper. Unpublished data on copepods recordedRom ballast water obtained from ships in Coos Bay, Oregon, are cited with the permission of Dr JimCarlton as well as Jeff Cordell.

REFERENCES

Carlton, J.T. 1985, Transoceanic and interoceanic dispersal of coastal marine organisms: the biology ofballast water. Oceanography and Marine Biology Annual Review 23:313-371,

Chapman, J.W. 1988. Invasions of the northeast Pacific by Asian and Atlantic gammaridean amphipodcrustaceans, including a new species of Corophium J Crustacean Biology 8:364-382.

Chu, K,H,, P,F. Tarn, C H. Fung, and Q.C. Chen. 1997. A biological survey of ballast water in containerships entering Hong Kong. Hydrobiologia 352: 201-206.

Cordell, J.R., C.A. Morgan, and C.A. Simenstad. 1992. Occurrence of the Asian copepodPseudoCh'aptomus inopinus in the zooplankton of the Columbia River estuary J Crustacean Biology 12:260-269.

Cordell, J.R, and S,M. Morrison. 1996. The invasive Asian copepod Pseudodiaptomus inopinus inOregon, Washington and British Columbia estuaries. Estuaries 19: 629-638.

Elston, R. 1997. Pathways and Management of Marine Nonindigenous Species in the Shared Waters ofBritish Columbia and Washington. Puget Sound/Georgia Basin Environmental Report Series No. 5.Published by Puget Sound Water Quality Action Team, P.O. Box 40900, Olympia, Washington 98504-0900. 94 p.

Fleminger, A. and S.H. Krarner. 1988. Recent introduction of an Asian estuarine copepodPseudodiaptomus marinus Copepoda, Calanoida! into southern California embayments. Marine Biology98:535-541.

Gauthier, D. and D.A. Steel. 1996. A synopsis of the situation regarding the introduction ofnoiundigenous species by ship-transported ballast water in Canada and selected countries. CanadianManuscript of Fisheries and Aquatic Sciences No. 2380. 66 p.

Harbo, R.M. 1997. Shells and Shellfish of the Pacific Northwest. Harbour Publishing, Madeira Park,British Columbia.

Ichikawa, S�Y. Wakao, Y, Fukuyo. 1992. Extermination efficacy of hydrogen peroxide against cysts ofred tide and toxic dinoflagellates, and its adaptability to ballast water of cargo ships in Japanese!.Bulletin Nihon suisan-gakkai shi. 58 �2!:2229-2233.

Jamieson, G,S,, E,D, Grosholz, D.A. Armstrong, and R.W, Elner, 1998. Potential ecologicalimplications of the European green crab, Carcinus maerias, to British Columbia, Canada and Washington,U,S,A. J Natural History in press!.

Kelly, J.M. 1993. Ballast water and sediments as mechanisms for unwanted species introductions intoWashington State. Journal of Shellfish Research 12: 405-410.

Kim, LH. 1992. Invasion of foreign barnacles into Korean waters. Korean Journal of SystematicZoology 8: 163-175.

Piercey, G.E., C.D. Levings, M. Elfert, and R. Waters. 1998. Data record on ballast water sampling fromBritish Columbia ports, December 1995 to January 1997. Can. Data Rep. Fish. Aquat. Sci. in prep!.

Quayle, D,B. 1964, Distribution of introduced marine mollusca in British Columbia waters. Journal ofthe Fisheries Research Board of Canada 21.1155-1181.

Ruiz, G.M., J.T. Carlton, E.D. Grosholz, and A,H, Hines, 1997. Global invasions of marine and estuarinehabitats by non-indigenous species: mechanisms, extent, and consequences. American Zoologist 37'.621-632.

Vancouver Port Corporation. 1997. Harbour Master Department Standing Order � Ballast Water.February 1997. Available: Vancouver Port Corporation, 1900 Granville Street, Vancouver BC V6C 2P9.

LIST OF FIGURES

Figure 1 Chart of the Pacific Ocean showing ports where ballast water was taken on closed circles! andports in British Columbia where ballast water was sampled, Insert table shows the names of the variousports.

Figure 2 Monthly mean temperature of ballast water samples obtained from ships in Vancouver mean,se; n indicates number of observations!.

LIST OF TABLES

Table 1 Origin of ballast water and number of samples collected from vessels arriving in BritishColumbia ports.

Table 2 Number of individuals of major taxa obtained in 67 samples of ballast water &om deep sea shipsarriving in British Columbia ports, December 1995 to January 1997.

Table 3 Percent occurrence presence or absence! of selected invertebrate taxa and their origin in ballastwater froin three major regions of the North Pacific.

Table 4 bundance {number rn '! of all organisms in the ballast water survey, categorized by month ofsampling and origin of the ballast water. Data are nuinber m ', standard error, and number of samples!.ns indicates not sampled.

Table 5 Abundance of cyclopoid and calanoid copepods number m ! in 64 pump samples &om ballastwater obtained from three major regions of the north Pacific.

g~E~' Vl 3h

~[>g,%%~

Er!

IM

4 ChE~

! !I! r 4

I !

! IEIpmIJ

r,r

E

' I+;' ,ir.';E',;,

!~aSem EII

!

0Ire ~ i~

!'

r

' ~ !'!

118

03!

EB

ICI

Irr!errE"NI

QE

Cl

E"

Monthl Mean Tem eratures in Ballast Water of Vessels Sam led inVancouver Harbour December 1995 to Janua 1997

25;----

Dec Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec Jan95 96 97

Month

Figure 2 Monthly mean temperature of ballast water samples obtained from ships in Vancouver mean, se; n indicates number of observations!.

Table l Origin of ballast water and number of samples collected from vessels arriving in BritishColumbia ports.

' Indicates mid-Pacific or rnid Pacific/mixed origin for ballast water

InclUdes two samples where ballast water was taken on east of Japan

Indicates Northeast Asia origin for ballast water

' Indicates northeast Pacific/North American origin for ballast water

120

Table 2 Number of individuals of major taxa obtained in 67 samples of ballast water from deep seaships arriving iu British Columbia ports, December 1995 to January 1997.

NUMBER FOUND PERCENT OF TOTALINVERTEBRATE GROUP

TOTAL

Cnidaria Hydra spp!Medusa

Nernatoda

Roti fera

Polychaeta larvae!Bivalvia larvae!GastropodaCladocera

Ostracoda

Copepoda naupliiCopepoda copepoditesCalanoida

CyclopoidaHarpacticoidaMonstrilloida

Copepoda unaccountedCrustacean naupliiCyprid larvae

Amphi podaiso podaCumacea

MysidaceaDecapoda prezoeaDecapoda zoeaDecapoda megalopaAcarina

Collembola

ThysanopteraChaoboridae

Trochophore larvaeChaetognathaLarvacea

1

156

46

14

1179

180

109

5651

49

40

162

8859

8679

]665

1

827

7231

53

104

8

1

4

12

6

5

3

1

1

1

139

164

13

35364

<0.01

0.44

0.13

0.04

3.33

0.51

0.31

15.98

0,14

0.11

0,46

25.05

24,54

4.71

<0.01

2.34

20.45

0.15

0.29

0.02

<0.01

0.01

0.03

0,02

0.01

0.01

<0.01

<0.01

<0.01

0.39

0.46

0.04

Table 3 Percent occurrence presence or absence! of selected invertebrate taxa and their origin inballast water from three major regioiis of the North PaciTic. Note: includes two sainples obtainedby plankton net.

Mid-Pacific Mixed

�6 samples!Northeast Asia �0

sainples!Northeast Pacific

�1 samples!Organism

122

Medusae

Polychaete larvae

Bivalve Larvae

Gastropoda Larvae

Cladocera

CopepoditesCalanoid copepods

Cyclopoid copepodsCrustacean naupl ii

Decapod prezoea larvae

Decapod zoea larvae

Decapod inegalopa larvae

Trochophore larvae

6

31

19

6

25

19

69

75

50

0 6 0 0

8

30

20

13

10

5

73

75

43

0 3 3 8

0

27

18

18

18

18

82

9t

82

9 0 9 0

Table 4 Abundance number m ! of all organisms in the ballast water survey, categorized bymonth of sampling and origin of the ballast water. Data are number m, standard error, andnumber of samples!. ns indicates not sampled.

175 90,2! 906 878,3! 692�18,2! 1161�76,9! ns 2826�775,3! 634�97,3!ASIA

2 -,1!1082�89,3! nsns 779�24,2! ns

294 -,1!340 -,1! 737�12,3! ns nsns ns

NOV DEC JAN

1996 1996 1997

918�71,4! ns 8 -,1!300�74,5! 9066 -,1! 7�,6!ASIA ns

2314�740,3! 803�48,3! 236�36,2! 159�49,2! ns 112 -,1!

5613�733,3! 2 -,1! nsns ns

123

MID PACIFICMIXED

NORTHEASTPACIFIC

MID PACIFICMIXED

NORTHEASTPACIFIC

DEC

1995

JULY

1996

JAN

1996

AUG

1996

FEB

1996

SEP

1996

MAR APR

1996 1996

OCT

1996

MAY

1996

JUN

1996

Table 5 Abundance of cyclopoid and calanoid copepods number m ! in 65 pump satnplesfrom ballast water obtained from three major regions of the north Facific.

TOTALMID PACIFICMIXED

NORTH EASTASIA

NORTHEASTPACIFIC

COPEPODA

24

2080

523

31676

3716

21043

4

2

19412

6

34

3790

307

761

0

110

0

11

0

0

7132

2

9

020

2149

04522

0

90

0

TOTAL 8661

~ indicates this species found in ballast water originating in Japan, Cordell, pers. comm.!

32

251

3790

850

60776

8348210

63

42

35205

sampled at Coos Bay, Oregon

124

CALANOI DA

Acartia sp.Acartia californiense

Acartia clausi

Acartia longiremisCalanus sp.Cal anus marshal lae

Calanus pacificus~Centropages abdominais*Clausocalanus 1 ividus*

Clausocalanus pergensCtenocalanus vanus

Diaptomus sp.Euchaeta sp.Eurytemora sp.Eurytemora pacificaMetridia sp.Metridia lucens

Metridia pacificaMicrocalanus pygmaeusNeocalanus cristatus

Paracalanus parvusParvocalanus crassirostris

Pseudocalanus sp,Pseudocalanus marinus*

Pseudocalanus newrnani*

P seudodiaptornus sp.Pseudodiaptomus marinusScaphlocalanus sp.Scolecithrice 1 la minorTortanus discaudatus

Undinel1 a sp.CYCLOPOIDA

Coryc acus sp,Corycaeus anglicus~Cyclops sp.Hemicyclops sp.Oithana sp.Oithona atl anti ca

Oithona brevicornisOithona similis

Oncaea sp.Oncaea borealis

Oncaea pro 1ata

0 012

776

0 5102

4 0 0 4 0 2 0 0 510 2 0 0

126

0

655

102

0 0110

2 04

29

10

0

8007

40

96

113

43

141

00

0

22

6

31

0

36

40

2

1849

4

901

0

28

0

141

02

014

0

7

1087

0

0

29

70

05

5

0

6860

0

00

0

00

0

97

0

0

0

32

300

0

650

10

7

9106

816

96

148

215

145

5

5

4

6884

6

315

10

3840

2

2072

4

1556

10228

32

2812

2

6943