Ballast Water Research at the WFRC 

Invasive aquatic species are considered to be one of the 

greatest threats to marine biodiversity, coastal economies, 

and even human health. Ballast water is a primary vector 

for these bioinvasions. One method of reducing risk of 

ballast water introductions is mid-ocean exchange of marine 

ballast water. However, it is widely recognized that this 

practice has many limitations, including serious ship safety 

concerns and variable biological effectiveness. 

International agencies and federal, state and local 

governments are attempting to tackle the problem by 

recommending or mandating the installation of ballast water 

treatment systems. The WFRC is working with others to 

develop new technologies to detect, prevent, and control 

aquatic invasive species impacts in Puget Sound and the 

Pacific Northwest.

Water is pumped into vast tanks to stabilize a vessel, and later the

ballast water is discharged. In this way, animals and plants, like

the copepod shown above, can be transported thousands of 

kilometers from their native habitats and deposited in a new 

environment.

WFRC Studies Ballast Water 

Although there are several vectors for invasive marine 

species such as aquaculture, recreational boats, and public 

aquaria, the most important transportation vector is ballast 

water carried in commercial ships. Today there are more 

than 45,000 commercial cargo-carrying vessels that use 

ballast water to maintain stability when cargo loads vary. 

Puget Sound is home to major seaports of national and 

international importance. Elsewhere in the Pacific 

Northwest, such as the Columbia River, significant ports are 

located far inland (e.g., Portland, Oregon) or in other 

estuarine settings. The region’s strategic importance in

Asia-Pacific trade makes it exceptionally vulnerable to 

ballast water introductions.

One ship may contain more than 12 million gallons of 

ballast water providing a very efficient vector for aquatic 

microbes, plants, and animals. In fact, it is estimated that 

7,000 different species are transported around the world 

daily in ballast water. Therefore, there is a critical need to 

develop tools to detect and eradicate invasive species in 

ballast water prior to their release and establishment in 

coastal ecosystems. At WFRC, scientists are working to 

develop DNA markers to detect and monitor invasive 

species in ballast water. These molecular tools offer great 

promise as a rapid assessment technique to evaluate the 

effectiveness of oceanic exchange which occurs off the 

West Coast for domestic shipments and across the NE

Pacific for international carriers. Using state-of-the-art

technology, we are developing diagnostic systems that will 

allow as many as 20,000 species to be detected at one time. 

This technology will give state and federal regulators the 

tools they need to monitor how clean ballast water is before 

it is dumped in coastal ports. Additional analyses of ballast 

water samples collected in Puget Sound are underway at the 

WFRC for quantification of bacterial and phytoplankton 

pre- and post eradication applications. 

Prevention and Control of Marine Ballast Water 

Organisms. 

For the past 4 years, the WFRC and University of 

Washington partners, have performed ballast water 

treatment experiments at the USGS’s, Marrowstone Marine 

Field Station in northern Puget Sound. Intermediate scale, 

75-gallon (280L) circular fish tanks, were utilized for the

Challenges for Treatment Technologies 

• Kills the toughest organisms. 

• Environmentally friendly

o Discharge should be nontoxic to the most 

sensitive bioassay organism that 

environmental agency chooses. 

• Safe for crew. 

• Noncorrosive to ballast tanks. 

• Relatively inexpensive. 

• Verification, certification, enforcement.

U.S. Department of the Interior USGS FS 2006-3080 

U.S. Geological Survey May 3, 2006 
ballast experiments. Experimental treatments and controls 

were replicated by using four tanks per replicate. The tank 

volume was sufficient to allow re-sampling four or more 

times during an experiment. Thus far, we have examined 

the effects of exposure of ultraviolet light, SeaKleen®, 

ozone, and electrolytically generated sodium hypochlorite 

on the survival of planktonic organisms characteristic of 

Puget Sound. The biological parameters measured included 

bacteria numbers, heterotrophic bacteria (deriving 

nourishment from organic substances), chlorophyll a, 

phytoplankton, and zooplankton. For the experiments 

oxidizing biocides, like ozone and disinfection byproducts 

were measured. 

Highlights of Experimental Results Include: 

Summary of UV Results 

Bacteria and phytoplankton more sensitive to UV than 

zooplankton.

• 50 – 75 mJ cm-2

yields a 4 or more log reduction of 

viable bacteria. 

• 300 mJ cm-2 necessary for rapid death of 

zooplankton.

Lethality of UV may not be instantaneous for larger taxa. 

Sub-lethal doses may induce stress effects (e.g. activity, 

spawning). 

Research Results from Experiments with Filtration and 

Chlorine are Most Promising. 

Within 5 hours post treatment: 

• filtration reduced zooplankton numbers by 83-89 

percent. 

• 1.0-1.5 ppm chlorine decreased zooplankton 

abundance 87-93 percent. 

• 0.5 ppm chlorine with filtration killed most of the 

zooplankton.

Immediately after filtration + chlorination, bacterial levels 

dropped by more than 50 percent. However, over time 

bacteria rebounded and bacterial counts in treated groups 

were two times higher than in untreated groups at the end of 

the study. Filtration without chlorination was not effective 

at reducing bacterial counts. Filtration + chlorine was very

effective at reducing phytoplankton numbers; most of the 

removal resulted from the chlorine. (See sidebar) 

Detection of Invasive Species 

WFRC scientists are developing species specific DNAbased diagnostic systems to detect non-native species in 

ballast water and to study invasion pathways in Puget 

Sound. A community approach using microarrays and 

fluorescing molecular beacons was chosen because rapid, 

high fidelity techniques are necessary to analyze and 

identify all life stages of marine organisms that may be 

carried in ballast water. A critical aspect of this tool 

development is converting the target DNA from a double 

stranded molecule to single stranded molecule so it may

bind to diagnostic beacons. To date, 24 species-specific 

diagnostic markers for native and non-native bivalves and 

crustaceans have been developed. Protocols to efficiently

extract DNA from planktonic life history stages are being 

developed. The extraction protocol will provide the 

sensitivity needed to detect as few as 250 larvae per water 

sample from ballast tanks or nature. 

By the end of 2006, the WFRC will have a complete 

collection of all native and introduced clams in Puget 

Sound. Diagnostic systems will be completed by winter 

and we will begin to coordinate with State managers to 

screen ballast water in ships entering Puget Sound. Because 

the ballast water invasive pathways are international in

scope, the WFRC is also acquiring biological samples from

a number of Pacific ports. 

The development of DNA markers from foreign ports will 

eventually allow agencies to determine the identity and 

origins of organisms present in ballast water. 

Live Zooplankton Per Liter 

Prototypic Treatment 

Technology at USGS 

Marrowstone Marine Field

Station. Three electrolytic 

cells are used to generate

hypochlorite from seawater

(NaCl). Each cell consists of 

a cathode and an anode. 

Filtration+Chlorine: Zooplankton 

Filtration + Chlorination
and to the Washington State Aquatic Nuisance Species

Committee of which the WFRC is a member. The WFRC is 

providing geospatial database services and GIS support to 

the Committee. 

Regional Testing and Evaluation Facility 

In March 2006, the WFRC received a start-up grant from the 

NOAA National Sea Grant and U.S. Fish and Wildlife 

Ballast Water programs to develop a full proposal for a 

research, development, test and evaluation (RDTE) facility

in Puget Sound. The full proposal will describe facility 

requirements necessary to conduct cooperative research on 

(1) ballast water treatment technologies; and the (2) biology 

and ecology of invasive species for the Pacific Northwest 

region, Oregon to Alaska. FY 2007 activities will include 

preparation of preliminary concept designs for: 

modifications and expansion of the Marrowstone Marine 

Field Station facility; modifications and development of the 

existing University Washington freshwater facility; and 

building a ballast water treatment test bed on a barge. 

Considerations will be given for either constructing a new 

barge or modifying an existing MARAD-owned barge. The 

possibilities and costs for using a barge will be compared to 

a land-based test bed alternative. The University of 

Washington, and the Glosten Associates Inc. are WFRC 

cooperators on this new project. 

Conceptual Model of Molecular Diagnostic Systems 

before hybridization after hybridization 

Testing an Environmental Sample 

How Molecular Beacons Work 

Shipboard Tests

(100's of cubic meters)

Testbed Experiments

(100's of cubic meters)

Mescocosm Experiments

(100's of liters)

Microcosm Experiments

(liters)

Benchtop Experiments

(milliliters to liter)

Suggested Model for Performing Ballast Water Treatment 

Research

Ideally, experiments of potential treatment technologies 

should begin with experiments being performed at scales 

identified at the top of the figure. The model demonstrates 

the need to have experimental capability at various scales 

including at an intermediate scale (which will be proposed 

for USGS at the Marrowstone Island facility. The 

Marrowstone facility will be an important site for vendors to 

have prototype treatment systems tested. 

Risk Assessment 

DNA-based detection tools are being used to analyze the 

distribution, establishment and ecology of invasive bivalves 

and crustaceans around Puget Sound. Geospatial data from

these analyses will be used to assess risk of invasion to 

native and commercial species. Invasive tunicates, green 

crabs, and Spartina are of special interest in Puget Sound 

Science Cooperators and Partners 

The WFRC’s science cooperators and partners in the Pacific 

Northwest include the University of Washington (School of 

Ocean and Fisheries, Biology Department, and Washington 

Sea Grant Program, Urban Waters Marine Research Center 

– Tacoma), University of San Francisco, University of 

Purdue, Washington Department of Fish and Wildlife, the 
Species found in Puget Sound 

ballast water. Meroplankton

are species that have planktonic 

early life history stages (eggs and

larvae) but adults dwell on the 

sea floor. 

Photos provided by Jeff Cordell.

technology to treat ballast water on ships. 

Appl. Environment. Microbiol.

Hershberger, P.K., S.A. Hart, J. Gregg, N.E. 

Elder, and J.R. Winton. In Press. Dynamics of 

viral hemorrhagic septicemia, viral erythrocytic 

necrosis, and Ichthyophoniasis in juvenile

Pacific herring. Diseases of Aquatic 

Organisms. 

Herwig, R.P., J.R. Cordell, S.M. Bollens, J.L. 

Grocock, N.C. Ferm, G. Rollwagen-Bollens, 

A. Slaughter, and J. Fulmer. In press. Effects 

of UV radiation on marine plankton

communities: implications for ballast water 

treatment. Mar. Eco. Prog. Ser. 

Cangelosi, A., and N. Mays. 2004. Summary

and Findings of the Ballast Discharge 

Monitoring Device Workshop, Marrowstone 

Marine Field Station, Olympic Peninsula, 

WA, USA, August 12-16, 2002. NortheastMidwest Institute, Washington D.C. 

biology, and toxicity. 

Mar. Ecol. Prog. Ser. 

Paulson, B.A., E.R. 

Blatchley III, S. 

Chaiyapechara, and R.P. 

Herwig. In Press. Effect 

of ultraviolet light on 

marine heterotrophic 

bacteria: evaluation of a 

U.S. Navy, U.S. Fish and Wildlife Service, 

Environmental Protection Agency, State of 

Alaska, National Oceanic and Atmospheric 

Administation (Sea Grant, NOAA Fisheries, 

and National Ocean Service), Department of 

Homeland Security– U.S. Coast Guard, 

Maritime Administration, Northeast Midwest 

Institute, and Smithsonian Environmental 

Research Center. Other cooperators and 

partners include the port authorities for Seattle 

and Tacoma, Puget Sound Action Team, 

Battelle Marine Sciences Laboratory, BP 

Exploration, Prince William Sound Regional 

Citizens Advisory Committee. 

Selected Science Products 

Herwig, R.P., J.R. Cordell, J.C. Perrins, P.A. 

Dinnel, R.W. Gensemer, W.A. Stubblefield, 

G.M. Ruiz, J.A. Kopp, M.L. House, W. J. 

Cooper. 2006. Ozone treatment of ballast 

water on the oil tanker S/T Tonsina: chemistry, 

Species found in Puget 

Sound ballast water. 

Holoplankton are 

planktonic species that are 

pelagically distributed 

through the whole of their 

life cycles. 

Photos provided by Jeff

Cordell.

For Additional Information, Please Contact: 

Lyman Thorsteinson, Center Director Phone: (206-526-6282

U.S. Geological Survey FAX (206) 526-6654

Western Fisheries Research Center http://wfrc.usgs.gov

6505 NE 65th Street 

Seattle, WA 98115