Protocols for Monitoring Pfiesteria and Related Health and Environmental Conditions in U.S. Coastal Waters

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U E .S . C D R E E P M A M RT O MENT OF C National Ocean Service National Centers for Coastal Ocean Science Center for Coastal Monitoring and Assessment Protocols for Monitoring Pfiesteria and Related Fish Kills and Environmental Conditions in US Coastal Waters by Donna D. Turgeon, Jamison L. Higgins, Danielle Luttenberg, Harris White, Marcia Orencia, Nancy Craig, and Karen Bushaw-Newton.

-EXECUTIVE SUMMARY- State and Federal agencies need comparable and reliable data to detect bloom development and improve the ability to forecast bloom formation of species within the toxic Pfiesteria complex (TPC: see Glossary) in US coastal waters. NOAA’s National Ocean Service held two workshops that brought together resource managers and scientific experts from Federal and State agencies and academic institutions to seek consensus on standard protocols for monitoring fish kill events that may have been caused by toxic Pfiesteria piscicida and for routine monitoring in waters known to support Pfiesteria and Pfiesteria-like organisms. The first, a Workshop to Standardize Pfiesteria Monitoring Protocols, was held 14-15 December 1998. The second, a Workshop to Standardize Fish Health Monitoring Protocols at Suspected Pfiesteria Events, was held 22-23 June 1999. The objective for both workshops was to establish common procedures for collecting and analyzing samples. Deliberations by the more than 50 invited specialists on harmful algae, fish pathology, and other disciplines, including several experts on Pfiesteria, resulted in a set of recommendations for a multidisciplinary, monitoring and assessment program. Workshop participants agreed to make agency data sets available for an integrated electronic database that would support regional and national assessments and recommended the following. 1. A three-tier, national program to monitor water quality, fish health, and phytoplankton with concomitant goals and strategies for:

• Rapid Event Response monitoring to be done by agencies during or just after significant fish kill events that may be related to toxic strains of TPC species; • Comprehensive Surveys and Assessments to be carried out in areas known to have supported toxic Pfiesteria outbreaks and in areas deemed to be at high risk for toxic events; and • Routine Monitoring of water quality, fish health, and phytoplankton to be conducted in areas that could support toxic strains of TPC species.

2. That phytoplankton, fish health, and a suite of water quality parameters be monitored simultaneously at sites known to support Pfiesteria-like species or sites at risk for toxic strains of Pfiesteria piscicida.

3. A set of standard parameters and methods for monitoring ambient water quality conditions, plankton, and fish health at sites suspected of having toxic strains of Pfiesteria.

The following report integrates the recommendations from both workshops into a set of standard protocols that could have substantial and lasting benefit to the nation’s understanding of harmful algal bloom dynamics and mitigation of their impacts in coastal waters. States with Pfiesteria concerns held training courses and implemented these protocols in 1999. INTRODUCTION waters. That same year, the natural resources and the local economy in Pfiesteria and Harmful Algal Maryland were also threatened when a Bloom Problems. Harmful algal fish kill of 30,000 was associated wih a blooms, HABs (see Glossary: HAB), toxic Pfiesteria outbreak in three regularly threaten US coastal living Chesapeake tributaries on the bay’s resources, restrict local harvests of fish Eastern Shore. and shellfish, divert public funds to health and environmental monitoring Recently, JoAnn Burkholder and programs, and depress local recreational Howard Glasgow reported a second and service industries. HABs occur Pfiesteria species, Pfiesteria naturally in our coastal waters, but their shumwayae sp. nov. (Glasgow, 2000), frequency, intensity, and distribution that also has ichthyotoxic properties. At appear to be increasing. Blooms of present, these are the only two species familiar and previously unknown with demonstrated ability to produce species have occurred in coastal areas, bioactive compounds that cause fish and HABs have now been identified in distress, disease, and death, in the toxic almost every coastal State from the Gulf Pfiesteria complex, TPC (see Glossary: of Maine through the Gulf of Mexico TPC). Studies are underway to test if a and north to Alaska. A recently discov- number of other, yet unnamed, Pfieste- ered HAB species, the dinoflagellate ria-like organisms, PLOs (see Glos- Pfiesteria piscicida, was first identified sary: PLO), found in East Coast tidal in 1988 in fish cultures (Smith et al., areas have similar icthyotoxic character- 1988), and in 1991 at a fish kill in the istics. Pamlico Estuary in North Carolina (Burkholder et al., 1992). P. piscicida Additionally, dead and dying fish has been implicated in the death of collected during PLO events often have millions of fish skin ulcers that in the nation’s A supermarket featured in 1997 this sign that harbor a second largest sought to ease customers’ fears of Pfiesteria. pathogenic and mainland highly invasive estuary, the NOTICE TO OUR Aphanomyces Albemarle- CUSTOMERS: fungus associ- Pamlico, with ated with an Some concerns have arisen toxic outbreaks regarding the waters of the intense granulo- occurring nearly Pokomoke River along the matous cellular every year since Maryland-Virginia border. inflammatory 1991 Presently, we are not selling response. In (Burkholder & any fish or shellfish harvested controlled Glasgow, 1997). from this river. laboratory During 1997, a experiments, P. fish kill of 1.2 piscicida semi- million fish was purified toxin linked to a toxic has been shown Pfiesteria to destroy fish outbreak in skin so that the North Carolina fish become

1 vulnerable to attack by opportunistic Mexico States. Additionally, a substantial fungi and bacteria, and open bleeding program of research on the TPC and sores develop. Research is underway to Pfiesteria look-alike species has been resolve the cause of such sores or ulcers added to the multi-agency federal in estuarine conditions, and the possible program that studies the Ecology and interactions of TPC species, other HAB Oceanography of Harmful Algal Blooms species, and various bacterial and fungal (ECOHAB). species in lesion formation and development. Finally, coastal regions that experience periodic HAB outbreaks know first hand The National Oceanic and Atmospheric that immediate monitoring and assess- Administration (NOAA), along with the ment of an event is but the first step in a longer process needed to effectively Maryland Fish and Seafood Tips— control and manage resource damage and

A HEALTHY FISH PFIESTERIA FACTS When purchasing fish, look for signs of freshness and quality: A mild scent Food and Drug Administration scientists have found no evidence of Pfiesteria toxins in seafood, even when it was taken from waters where the microbe was blooming.

FDA scientists have examined crabs taken from the Pocomoke River after the toxic Pfiesteria outbreak there, as well, as oyster’s Scales that adhere placed in tanks with toxic Pfiesteria. Bright, clear eyes tightly to skin In both cases, no toxins were found in the edible parts of the animals. Bright pink or red gills Skin that springs back when slightly pressed For information on how to handle Do not eat fish or Do not harvest or eat fish or shellfish from Some sores, injuries, and discolorations in fish and prepare Maryland seafood, shellfish with sores or areas where fish are dead or dying. are normal from contact with fishing gear and contact the Maryland Department lesions or fish that Maryland does not require sellers to label predators. Such fish are routinely discarded by of Agriculture’s Seafood appear unhealthy. the source of fish, but buyers should ask if fishermen and dealers before reaching the Marketing Program at 410-841- market. But when in doubt, throw it out. they are concerned. 5820.

In response to consumer fears about Pfiesteria piscicida and the safety of Chesapeake Bay seafood, state and federal officials offer safe-seafood guidelines. Environmental Protection Agency (EPA), threats to the public. For these reasons, the Centers for Disease Control and agencies must continue to sponsor long- Prevention (CDC), the United States term programs to monitor water quality, Department of Agriculture (USDA), and fish health, and HABs, to analyze existing other Federal agencies responded in 1997 data, and to share results. to the Maryland outbreaks of Pfiesteria piscicida by providing immediate support Workshops for Gaining Consensus to supplement the State’s monitoring and on Monitoring Protocols. NOAA, response activities. This effort combined EPA, and the Interagency Pfiesteria Federal and State resources to monitor Working Group agreed there was a need environmental conditions and assess to standardize monitoring protocols immediate watershed land use and among the many agencies that collect nutrient loadings as potential contributing information on water quality, fish health, factors to fish kills. The Federal govern- plankton, and environmental conditions ment is continuing its partnership with the associated with PLO events. NOAA States by providing funding for Pfiesteria proposed holding a workshop to gain piscicida monitoring and assessment consensus among State and Federal efforts in coastal Atlantic and Gulf of agency representatives and scientific

2 experts. NOAA’s first Workshop to Stan- for a coordinated national response to dardize Pfiesteria Monitoring Protocols was toxic HAB outbreaks. The workshop held December 14-15, 1998. A second focused on monitoring phytoplankton, Workshop to Standardize Fish Health fish health, and water quality. Over the Monitoring Protocols was held June 22-23, course of two days, there were three two- 1999. Conducted by the Center for Coastal hour working sessions, one for each topic, Monitoring and Assessment (CCMA), one during which three break-out groups of the National Centers for Coastal Ocean independently explored the issues related Science (NCCOS), both workshops were to the topic and developed recommenda- convened in Silver Spring, MD. tions through facilitated discussions. Experts introduced each topic and set the Workshop goals were three-fold. First, reach stage for the break-out group deliberations agreement on protocols for rapid-response that followed. Patricia Glibert made an assessments of toxic Pfiesteria outbreaks. oral presentation on measuring water Second, recommend a suite of standard quality parameters relevant to PLO, parameters that should be measured (e.g. Wolfgang Vogelbein spoke about water quality, fish health, and phytoplank- assessing fish health, and JoAnn ton) when responding to events. Third, Burkholder presented information on field discuss the integration of state and federal sampling and phytoplankton analytical agency data sets for regional and national protocols. Supporting materials that were assessments. This report presents the provided to participants for this workshop deliberations of participants from both included information on the water quality workshops. parameters that were then being monitored by State agencies and on how and in Workshop I. Coastal state administrators what electronic formats their data were from New Jersey through Texas were asked stored. to designate two state experts who had responsibility for monitoring water quality, After each two-hour working session, the fish health, and phytoplankton. Representa- facilitators (Water Quality - Patricia tives from other Federal agencies with Glibert, John Pennock, and Tracy interest in these topics also attended, i.e., the Villareal; Fish Lesions/Mortalities – Environmental Protection Agency (EPA), Andrew Kane, Mac Law, and Helen the Centers for Disease Control and Preven- Schurz-Rogers; Phytoplankton – Donald tion (CDC), the Food and Drug Administra- Anderson, Howard Glasgow, and Karen tion (FDA), the US Department of Agricul- Steidinger) summarized their break-out ture (USDA), and the US Geological Survey deliberations to the full assembly. On the (USGS). Of the 51 attendees, 21 were from last day, all of the presenters, facilitators, State agencies, 21 from Federal, and 9 from and discussion recorders (Karen Bushaw- universities and other non-governmental Newton, Nancy Craig and Danielle organizations (see Appendix 1 for the list of Luttenberg), assembled a single set of attendees of Workshops I and II ). recommendations for each topic. The integrated recommendations were then Workshop I opened with Bud Cross of reported to all participants in plenary NOAA and Craig Vogt of EPA reporting on session. Their recommendations were their agencies’ current efforts related to unanimously accepted as presented. monitoring and research, as well as the Federal Event Response Plan for Harmful Workshop II. At the first workshop, Algal Bloom Events—an interagency plan the fish health break-out groups con-

3 WHEN FISH ARE PRESENT IN PFIESTERIA WATERS... AND WATER IS BRACKISH, CALM, AND COLD AND WATER IS BRACKISH, CALM, AND WARM (about 12-15 degrees Celsius) (usually 26 degrees Celsius or higher)

TOXIC ZOOSPORES EXCRETA FISH LESIONS EXCRETA

GAMETES

PLANOZYGOTE NATURAL DEATH NONTOXIC ZOOSPORES

FISH KILL

CYSTS AMOEBAE

This artist’s depiction of the complex Pfiesteria piscicida life cycle includes cysts, amoebae, and toxic zoospores that may cause massive fish kills in coastal waters. cluded that a second workshop was Law made a presentation on fish collection, needed to specifically focus on the Andy Kane on laboratory analyses, and Joe parameters and protocols needed to Macknis on data management. monitor fish health related to Pfiesteria events. The experts from the first RECOMMENDATIONS FROM workshop were asked to provide a list of WORKSHOPs I & II candidate invitees for this focused fish health workshop. The resulting list Strategies for an Interagency included fish pathologists and/or field Pfiesteria Monitoring Program. personnel from the states that had existing Participants at both workshops recom- Pfiesteria programs, as well as academi- mended that water quality, fish health, and cians and representatives from Federal phytoplankton be monitored simultaneously agencies (e.g., EPA, FDA, USDA, at sites suspected of supporting problematic USGS). PLO. Participants outlined the following three-tier interagency program to monitor for Over the course of one and a half days, Pfiesteria and Pfiesteria-like organisms: there were three two-hour working sessions, one for each of the following 1. Rapid Event Response topics: fish collection procedures; 2. Comprehensive Surveys and laboratory analyses; and data storage, Assessment management, and sharing. Experts 3. Routine Monitoring introduced each topic and set the stage for the group discussions that followed. Mac

4 Participants proposed a suite of standard concern, then samples should be split and parameters for monitoring ambient water analyzed by two different laboratories to quality conditions at sites at risk from enable cross-corroboration of findings toxic strains of Pfiesteria. They agreed to about the presence of actively toxic strains make agency data sets available for an of TPC species and, potentially, newly integrated electronic database that would detected toxic PLOs. It is extremely support regional and national assess- important that toxicity be verified using ments. Workshop participants reviewed fish bioassays because this is the only and commented on this report. technique at present that can be used to verify the presence of actively toxic strains 1. Rapid event response monitoring of TPC species. done by state agencies during fish kills or lesion events that may be related to Water Quality. Water samples should be PLOs. filtered. Minimally, measure the following: • Station latitude and longitude Goal: Characterize water quality, • Date and time fish health, and the phytoplankton • Tidal stage and water depth community at sites of fish kills or • Weather conditions fish lesion events that may be due to • Current speed and direction toxic Pfiesteria activity. • Light penetration/Secchi disc/ turbidity When responding to a potential ongoing • Temperature (±1o Centigrade) toxic Pfiesteria outbreak, safety precau- • Salinity (± 0.1 ppt) tions including protective clothing and • pH (± 0.2) respirators should be used until all • Dissolved oxygen (± 0.5 ppm) samples are sealed and decontaminated • Dissolved ammonia (e.g., North Carolina’s prescribed • Dissolved organic nitrogen, carbon, protocols). Nitrile gloves (not latex) and phosphorus should be worn when touching the • Dissolved nitrate plus nitrite equipment, water, and bottles. Bottles and • Dissolved phosphate equipment can be decontaminated fairly • Dissolved silicate well by washing in a dilute Clorox® • Chlorophyll a solution. If possible, samples for monitoring should be collected along transects outward from the epicenter to beyond the Rapid response assessment during a kill zone. In addition to sampling suspected bloom of Pfiesteria piscicida. phytoplankton, to confirm the presence of toxic dinoflagellates, water samples should be collected for different types of bacterial, viral, and fungal pathogens; organic chemicals (pesticides and herbicides); and toxic blue-green algal species. Laboratory samples for presumptive counts and for fish bioassays should be analyzed “blind” to guard against bias. When presumptive counts (see Glossary: Presumptive Count) are positive for PLOs in concentrations high enough to be of

5 Fish monitoring. If possible, trained fish Fisheries Society procedures (AFS, health response personnel, including a 1982; Thoesen, 1994; Meyer & fish pathologist, should be on-site for fish Barclay, 1997). health events. Evaluate fish behavior, • On a randomly selected subsample of make overall Dead and lesioned fish from 100 fish affected per species, “map” environmental a toxic Pfiesteria event. on a diagrammatic fish the location, observations, type (i.e., loss of scales, ulcers, and take a reddened/ discoloration, raised photographic masses, normal, other), and preva- record of the lence of lesions. event and • Give each fish a unique accession typical fish number on a separate data sheet; the condition. At data sheet, photographs, and jars each site, should all be labeled with the same estimate the accession number. If possible, use a mortality and microscope in the field and examine prevalence of fresh fish for gill, skin, and gut lesioned fish parasites. for all species • For histopathological analyses, collected. collect a subsample of 25 fish per Subsample species (10 moribund lesioned, 10 diseased, fresh dead, and healthy fish for moribund normal, and 5 fish from pathogen analyses. Necropsy fish samples sites near but outside the kill and for histopathology, parasitology, and upstream). Either conduct a full field microbiology (bacteriology, mycology, necropsy on 20 fish (including virology): live/moribund* fish are bacterial analysis), a 3-minute preferred for these analyses but in the necropsy (see Appendix 2), or chop absence of live/moribund fish, fresh dead the whole fish into two or three fish* should be collected. Moderately pieces, depending on size of fish. A decomposed fish* can be used for serological collection should be done toxicological analyses only. Severely before the necropsy. Preserve the decomposed fish* are unsuitable for any organs for each fish in separate, analyses. labeled vials containing 10% buffered formalin (10 parts formalin Live fish should be euthanized with to 1 part fish). MS222 or by severing the spine follow- • If there is a pathologist or other ing American Veterinary Medical trained personnel on site, then Association (AVMA) protocols (J. Am. histological and microbiological Vet. Med. Assoc., 1993). Archive tissues samples should be field-processed and blood and make a photographic and preserved with appropriate record of pathologies. fixative and microbiological trans- • To determine the geographic extent port media for further laboratory of the fish kill, use American study. For viral assays, dissect liver,

* Criteria for selecting fish for histological, microbiological, virological, and toxicological analysis are based on gill and body coloration, integrity and odor: (1) Live/Moribund (gills firm, body color vibrant, tissue firm, no odor); (2) Fresh Dead (gills firm, body color/markings still apparent, tissue firm, no odor); (3) Moderately Decomposed (gills pale pink, body color/markings faded, tissue spongy, slight odor); and (4) Severely Decomposed (gills white, body color/markings indistinguishable, tissue mushy, strong odor).

6 kidney, spleen (excluding samples preserved with acid Lugol’s intestine) and/or muscle lesions solution. and store in Hank’s Balanced Salt • Conduct fish bioassays with fresh Solution with 10% fetal calf samples to test for toxicity; if serum, 500 IU/ml of penicillin, positive, then use scanning electron and 500 mg/L of streptomycin and microscope (see Glossary: SEM) on ice or refrigerate. These samples specially prepared cells (suture- must be assayed within 72 hours. swollen or membrane-stripped) to In the event there is no pathologist identify the dinoflagellates to to conduct field processing, then species. individually bag (in plastic) and • Perform algal assays for information immediately place on wet ice on other mixotrophic/heterotrophic another 25 moribund fish and 10 dinoflagellate species present that normal fish for laboratory eat algal prey. Among these, the bacteriological and virological PLO species that have increased in analyses. If the fish to be used for abundance in response to the histology are also to be used for presence of this algal food have all microbiological analyses, then been found, thus far, to be incapable collect an additional 15 moribund of producing bioactive compounds fish. It is imperative to deliver in enough concentration to harm or iced samples to the laboratory for kill fish. analysis within 24 hours. Micro- • In the future, as assays become biological analysis should be available that can be used reliably to conducted according to the detect TPC species toxins, such procedures outlined in the Fish assays should be used to confirm the Health Section of the American presence of TPC toxins. Also, Fisheries Society “Bluebook” whenever possible, use the molecu- (Thoesen, 1994). lar probes that have been tested and • For toxicological analyses, collect found reliable to detect the two TPC and flash-freeze 25 fish. For species. Samples (water and retrospective analyses, archive 25 sediment) should be taken for (10/10/5) whole fish in a freezer analysis by probes. from each kill. 2. Comprehensive surveys and assessments conducted in areas known to Phytoplankton Monitoring. During an have supported toxic Pfiesteria out- in-progress fish kill or fish lesion event, or when fish are acting erratically Biohazard III facilties are used at without signs of disease, collect NCSU when conducting fish bioassays phytoplankton samples from the with Pfiesteria. immediate vicinity and preserve with acid-Lugol’s solution for presumptive counts of PLO’s. Additionally, collect enough fresh, unpreserved water to conduct fish bioassays, algal assays (see Glossary: Algal Assay), and for molecular probe identifications. • Complete presumptive counts by light microscopy of plankton

7 breaks, and in areas deemed at risk for minimum of three times a year. Make a such outbreaks. Comprehensive survey photographic record of significant patholo- and assessment data are of particular gies, then archive tissues and blood. value in guiding future management efforts. Phytoplankton Monitoring. Follow the same procedures recommended for a Rapid Goal: Determine and predict the pres- Response to a toxic outbreak. ence and distribution of potentially toxic strains of Pfiesteria and Pfieste- 3. Routine monitoring conducted in areas ria-like species. that could support toxic strains of TPC species, especially where the water quality Monitoring should be conducted at conditions may be conducive to blooms of carefully selected sites (i.e., historically potentially toxic Pfiesteria and Pfiesteria- known fish kill/disease areas; quiet, like species. This broad-scale monitoring nutrient-rich waters in deposition areas can enhance existing State monitoring with organic sediments) during late activities. Note: greater efficiency, enhanced summer and early fall. communication, and better coverage could be achieved by integrating the efforts of state Species identification is confirmed through agencies and academic specialists who scanning electron microscopy (SEM). currently monitor conduct such activities independently.

Goal: Characterize the long-term dynamics of phytoplankton communities, indicate the presence/absence of TPC and PLO species, and determine their relation- ship to other members of the phyto- and zooplankton communities, water quality, and fish health.

Water quality monitoring. Measure the following set of physical and chemical water Water Quality. Make the same measure- quality parameters at all sites when monitor- ments that are recommended for a Rapid ing for suspected PLO: Response to a toxic outbreak. • Station latitude and longitude • Date and time Fish Monitoring. Follow the same procedures recommended for a Rapid Response to Fish health problems like these lesions may a toxic outbreak but, in addition, conduct indicate the presence of TPC organisms. routine monitoring pre- and post-season based on historical observations of problem times/areas. Depending on state or local needs and concerns, additional comprehensive sampling studies can be adopted. Collect fish samples from each site and necropsy for histopathology, microbiology, parasitology, bacteriology, mycology, and virology. Sampling should take place a

8 • Tidal stage and water depth • General weather conditions, wind speed • Current speed and direction • Light penetration/Secchi disc • Water temperature (±1o Centigrade) • Salinity (± 0.1 ppt) • pH (± 0.2) • Dissolved oxygen (± 0.5 ppm) • Dissolved ammonia • Chlorophyll a Buoys can be used to record in situ environmental information related to HABs. Fish monitoring. Note abnormal fish behavior, general health condition, and the Authority’s quality assurance frequency and type of external anomalies protocols) for plankton quantifica- (e.g., reddened spots on the skin, lesions) tion (include picoplankton but do not when routinely monitoring for fish health miss larger forms). Note that the at sites that might support toxic strains of Utermöhl method generally is not Pfiesteria and Pfiesteria-like species. acceptable for quantifying • Enhance existing state programs that picoplankton, so that other tech- routinely monitor coastal waters for niques such as Palmer cells environmental quality, fish, and (UNESCO 1978a, 1978b) should be plankton populations by sampling used. fish populations with cast nets to • Quantitative probes that enable capture 1-2 inch fish (10-20 casting determination of the cell number attempts). Map fish lesion preva- present are being tested for Pfieste- lence and location. Collect a ria spp. As their use becomes more subsample of these fish (5 normal routine, complete counts should be and 5 abnormal fish), perform a 3- made for Pfiesteria piscicida and minute necropsy (see Appendix 2) or other members of the TPC known to chop fish and preserve in 10% produce bioactive compounds that buffered formalin. hurt or kill fish. It is best to collect • Do histopathology processing when and process cells for DNA extrac- appropriate. tion: filter into a 2.5 cm glass fiber filter and immerse the filter in CTAB Phytoplankton monitoring. Collect buffer or some other solution integrated water-column samples (or designed for DNA extraction. pooled discrete depth samples), preferably Otherwise, a sample (50-100ml) at least in duplicate; or surface and bottom should be preserved with acid samples at each site routinely monitored Lugol’s solution and archived for for TPC and PLO species. later analysis. For rapid response •Characterize phytoplankton commu- events, DNA extraction is prefer- nity composition (to species level if able; for routine monitoring, the possible, if not, then to dominant second option may be acceptable. group and size class). However, PCR analysis of any • Use the Utermöhl method samples collected for Pfiesteria spp. (UNESCO 1978a, 1978b; i.e., with should be conducted within six Chesapeake Bay Program’s or months. Massachusetts Water Resources 9 Additionally, because Pfiesteria spp. tion, and link to Geographic Information have been found in the sediments at System applications. fish kill sites after the event, even when it has not been present in the CONCLUSIONS water column, collection and analysis of surface sediment material These Workshops filled a need expressed is also recommended. Small samples by both federal and state agency managers of sediment (1gm or cc) can be for consistent protocols to monitor assayed for the presence of Pfiesteria suspected toxic Pfiesteria outbreaks. More spp. using gene probes. However, than 60 managers and scientists who these probes cannot detect toxicity. participated in the two NOAA workshops Samples should be assayed as soon reached consensus on the need for as possible after collection as there is consistency in the parameters measured, as yet no established method for the analysis of samples collected by those sample preservation. States that have been monitoring over the past few years for Pfiesteria and PLOs, Integrated Data and Assess- and quality control/quality assurance ments. Both workshops included involving cross-corroboration of results by experts (Lowell Bahner, Bill Fisher, and Pfiesteria specialists with demonstrated Joe Macknis) who gave presentations on expertise in research on toxic strains of how to integrate now disparate agency TPC species. All recommendations put data sets into a national database of more forth by this group were agreed upon, reliable and accessible data. Participants including consensus on sharing agency monitoring data with other state and Routine water quality monitoring conducted at a site that could support Pfiesteria. federal agencies. This body of experts proposed a three-tier monitoring program: (1) rapid response to fish kill events, assisted by appropriate experts; (2) comprehensive surveys and assessments in areas that have experienced or are at risk for toxic Pfiesteria outbreaks (as well as other harmful algal blooms); and (3) routine monitoring at sites that might support toxic strains of TPC species. Attendees called for concurrent collection of phytoplankton, fish health, and water quality samples for each tier of their generally agreed that quality-assured data program. on this topic would be desirable for agency assessments and natural resource These recommendations will be valuable and human health decision-making. The for developing and improving rapid participants agreed that they would make response and environmental assessment at their data sets available and work towards sites suspected of suspected toxic assuring that spatial and temporal data Pfiesteria outbreaks, for state and federal were comparable if the national database monitoring of PLOs, and for comprehen- were to provide user-friendly access, sive surveys to determine if and where integrate with other databases (e.g., potentially toxic strains of TPC species sediment toxicity), facilitate summariza- may be present.

10 About NOS/NCCOS/CCMA

The National Ocean Service (NOS), a part of the National Oceanic and Atmospheric Administration (NOAA), approaches the 21st century with a clear vision — that our coasts and oceans enjoy robust health, provide a rich bounty of resources, and are wisely managed to endure competing uses.

NOS's National Centers for Coastal Ocean Science (NCCOS) works to understand the ocean and coastal environment, and the influences of human activities, by conducting research, monitoring, and assessments of these delicate and important areas.

NCCOS’s Center for Coastal Monitoring and Assessment (CCMA), moni- tors, surveys, and assesses coastal environmental quality, habitats, and resource distribution. The Center's National Status and Trends Program conducts long-term monitoring and bioeffects assessments of toxic contaminants at more than 350 coastal sites. The Center's Biogeography Program develops regional and national assessments on living marine resource distributions and ecology.

Credits: The Cover, and pages 5, 6, and 10 - the North Carolina Department of Environ- ment and Natural Resources; Opposite page, page 4 diagram, page 8 right photo, page 18, the first photo - J. Burkholder; page 1 - J.A. Purcell, The Washington Post; page 2 - The Washington Post; page 7 - North Carolina State University; page 8, left photo - Florida Marine Research Institute; page 9 - P. Glibert; page 17 - A. Kane; and page 18, the second, third, and fourth photos - K. Steidinger.

Proper Citation of this document is as follows: Turgeon, D.D, J.L. Higgins, D. Luttenberg, H. White, M. Orencia, N. Craig, K. Bushaw-Newton. 2001. Protocols for Monitoring Pfiesteria and Related Fish Health and Environmental Conditions in US Coastal Waters. NOAA, NOS,

With contributions from:

Mark Alling Joyce Evans Mary Ley John Pennock Donald Anderson Bill Fisher Michael Mac John Plumb Bill Andrews Howard Glasgow Joe Macknis Renate Reimschussel Larry Ausley Pat Glibert Harold Marshall John Schachte Kay Austin Dave Goshorn Pete Maye Helen Schurz Rogers Lowell Bahner Sherwood Hall Ron Michaels Kevin Sellner Joan Bernstein Wick Harlan Cynthia Moncreiff Harley Speir Vicki Blazer John Heisler Bill Muir Karen Steidinger Scott Brown Ken Hemphill Larry Needham Tracy Villareal JoAnn Burkholder Edythe Humphries Amanada Niskar Barbara Viskup Chris Caudle Andrew Kane Jimmie Overton Wolfgang Vogelbein Cindy Contreras Jan Landsberg Shelly Paraso Butch Younginer Cindy Driscoll Mac Law U.S. Secretary of Commerce Donald L. Evans

Acting Under Secretary for Oceans and Atmosphere and NOAA Administrator/ Deputy Under Secretary for Oceans and Atmosphere Scott B. Gudes

Acting Assistant Administrator for Ocean Services and Coastal Zone Management, National Ocean Service Margaret A. Davidson, Ph.D.

U.S. Department of Commerce National Oceanic and Atmospheric Administration National Ocean Service April 2001