[From the U.S. Government Printing Office, www.gpo.gov]





                                                                                                 WHOI-93-02




                          Marine Biotoxins and Harmful Algae:
                                               A National Plan




                                                            Donald M. Anderson
                                                             Woods Hole Oceanographic Institution
                                                              Woods Hole MA 02543
                                   Ilk
                                    till
                                                                 Sylvia B. Galloway
                               Ilya
                                                                  National Marine Fisheries Service
                                                                   Southeast Fisheries Science Center
                                                                    Charleston, SC 29422


                                                                       Jeanne D.Joseph
                                                                        National Marine Fisheries Service
                                                                         Southeast Fisheries Science Center
                                                                          Charleston, SC 29422







                                                     January 1993





                               Woods Hole Oceanographic Institution

                                                    Technical Report
   SH177          was providedoy the National Marine Fisheries Service Saltonstall-Kennedy Grant No. NA27FDO092-01,
                 tional Marine Fisheries Service's Charleston Laboratory and by the NOAA Coastal Oceans Program.
   .R4M37
   1993                                Approved for public release; distribution unlimited.
   c.2











                                                  WHOI 93-02

                                    Marine Biotoidns and Harmful Algae:
                                                A National Plan



                                               Donald M. Anderson
                                               Biology Department
                                      Woods Hole Oceanographic Institution
                                        Woods Hole, Massachusetts 02543

                                               Sylvia B. Galloway
                                        National Marine Fisheries Service
                                        Southeast Fisheries Science Center
                                        Charleston, South Carolina 29422

                                                Jeanne D. Joseph
                                        National Marine Fisheries Service
                                        Southeast Fisheries Science Center
                                        Charleston, South Carolina 29422


                                                  January 1993


                                               Technical Report



            Funding was provided by National Marine Fisheries Service Saltonstall-Kennedy grant No.
           NA27FDO092-01, National Marine Fisheries Service Charleston Laboratory and by the NOAA
                                            Coastal Oceans Program.

             Reproduction in whole or in part is permitted for any purpose of the United States Govern-
            ment. This report should be cited as: Woods Hole Oceanogr. Inst. Tech. Rept., WHOI 93-02.


                                          Approved for distribution:




                                        611bel C. GoldmaWChairman
                                             Biology Department



                           LIBRAR
                         NOAA/CC.FH
                     1990 HOBSON AV,',-,
                    CHAS- SC 29408-262c_









                                           TABLE OF CONTENTS


                                                                                          Page

            EXECUTIVE SUMMARY          .....................................             EI-E3

            PREFACE     ...................................................                i-iii


            1.   INTRODUCTION      ...........................................              I


            11. THE TOXINS      ..............................................              3
                 1. General Background     ......................................           3
                 2. Toxicology and Pharmacology   ................................          4
                    2.1 Background   ............................................           4
                    2.2 Impediments and Recommendations     ........................        6
                 3. Analysis, Standards, Chemistry  ...............................         8
                    3.1 Background   ...........................................            8@
                    3.2 Impediments and Recommendations     ........................        9

            III. BLOOM BIOLOGY AND ECOLOGY              ............................       11
                 1. Bloom Dynamics     .........................................           11
                    1.1 Background   ..........................................            11
                    1.2 Impediments and Recommendations     ........................       12
                 2. Phytoplankton Monitoring    ..................................         15
                    2.1 Background   ..........................................            15
                    2.2 Impediments and Recommendations     ........................       15

            IV. FISHERIES AND FOOD WEBS           ................................         17
                 1. General Background    .......................................          17
                    1.1 Shellfish: Impediments and Recommendations     ................    19
                    1.2 Finfish: Impediments and Recommendations      .................    23
                    1.3 Food Web Effects: Impediments and Recommendations       .........  24
                 2. Shellfish Monitoring Programs   ..............  I........   0 ........ 25
                    2.1 Background   .................................          % ........ 25
                    2.2 Impediments and Recommendations     ........................       27

            V. SUMMARY         ...............................................             30


            VI. ACKNOWLEDGEMENTS            ...........................         0..1 ...... 31


            VIL REFERENCES         ............................................            32


            VIH. WORKSHOP AGENDA            .....................................          39


            IX. PARTICIPANTS       ............................................            42



                                                     TOC-1













                      MARINE BIOTOXINS AND HARMFUL ALGAE: A NATIONAL PLAN




                                                 EXECUTIVE SUMMARY



              Marine biotoxins and harmful algae represent a significant and expanding threat to human health
              and fisheries resources throughout the United States. These phenomena take a variety of forms.
              From a public health standpoint, four human illnesses are associated with toxic algal blooms and
              consumption of toxin-contaminated shellfish in the United States: paralytic, neurotoxic, amnesic,
              and diarrhetic shellfish poisoning (called PSP, NSP, ASP, and DSP respectively). Except for
              ASP, all are caused by biotoxins synthesized by a class of marine algae called dinoflagellates.
              ASP is produced by another class of marine algae (diatoms) that until recently were thought to
              be harmless. A fifth human illness, ciguatera fish poisoning (CFP) is caused by biotoxins
              produced by epibenthic dinoflagellates attached to surfaces in many coral reef communities.
              Ciguatera toxins are transferred through the food chain from herbivorous reef fishes to larger
              carnivorous, commercially valuable finfish.        In a similar manner, the viscera of other
              commercially important fish such as herring or sardines can contain PSP toxins, endangering
              human health following consumption of whole fish. Whales, porpoises, seabirds, and other
              animals can be victims as well, receiving toxins through the food chain via contaminated
              zooplankton or fish.

              Marine fauna can be affected by a variety of algal species that release toxins or other compounds
              into the water or that kill by physically damaging gills. Problems associated with harmful algal
              species and farmed fish have increased considerably in recent years, due in part to the
              simultaneous expansion of the fish-farming industry. Furthermore, the death and decay of algal
              blooms can lead to anoxia through decompositional. oxygen demand, resulting in widespread
              mortalities of fish, shellfish, and invertebrates. An additional problem is that of benthic or
              planktonic macroalgae that can proliferate in response to anthropogenic nutrient enrichment. This
              can lead to major negative impacts due to displacement of indigenous species, habitat alteration,
              or oxygen depletion.

              The National Academy of Sciences recently issued a report expressing serious concerns about
              the quality of the nation's seafood (Ahmed, 1991), emphasizing the need for attention to marine
              biotoxins and harmful algae. In response to this directive and to a heightened public and
              governmental awareness of the changing nature of the coastal marine environment, governmental
              funding is being targeted towards marine biotoxins, harmful algae, and their impacts. The
              optimum allocation of these resources can benefit greatly from scientific guidance as the new
              programs are formulated and implemented. A workshop on Marine Biotoxins and Harmful Algae
              was thus convened in Charleston, South Carolina, from 21-24 April 1992, to bring scientists and
              regulatory officials together to evaluate U.S. research knowledge and capabilities, and to identify
              areas where research funds should be directed for maximum benefit.


                                                             E-I










                 From a number of nationally recognized leaders in the areas of marine biotoxins, harmful algae,
                 seafood safety, and public health, 24 participants were selected to represent the critical scientific
                 disciplines and all regions of continental North America. Position papers on 12 relevant topics,
                 written by the participants and distributed before the workshop, formed the basis for discussions
                 within the three working groups. Conclusions of the working groups were presented to all
                 participants in two plenary sessions.

                 Twenty-eight major impediments to progress were identified. These can be summarized as
                 follows:


                 Defteiencies related to the biotoxins: Toxin standards are largely unavailable; standard sample
                 preservation and handling protocols do not exist; existing assay methods are inadequate for
                 monitoring and research; molecular pharmacology and pharmacokinetics of marine biotoxins are
                 poorly understood; diminution or loss of toxin production can occur in laboratory algal cultures;
                 mass culturing of most toxic species is difficult.

                 Lack of information on harinful algae: Algal bloom dynamics and species succession are
                 complex and not yet predictable; the relative effects of natural versus anthropogenic influences
                 on population size, species composition, bloom longevity, and toxin production are unknown;
                 knowledge of the physiology of growth and toxin production is inadequate; toxin standards and
                 rapid assay methods are lacking; availability of isolates of toxic or harmful algae is limited.

                 Lack of information on impacted fisheries resources and protected marine resources: Toxin
                 uptake, metabolism, and depuration in shellfish, fish, and other marine animals is poorly known;
                 toxin sensitivities of different life history stages, and long-term effects of algal metabolites on
                 growth, reproductive success and recruitment are unknown; movement of toxins through the food
                 web is poorly understood; databases are inadequate and not readily accessible to potential users;
                 methods for rapid field assays of fish or sheRfish are lacking; toxin standards are often
                 unavailable; analytical methods for toxin detection in animal tissue need improvement.

                 Inadequate mechanisms and knowledge to protectpubfic healthfully: Early warnings of known
                 and unknown toxins are required to protect consumers and industry; assay methods need
                 improvement; toxin standards are not always available; sampling programs are inadequate for
                 bloom detection or characterization; the extent of seafood poisonings is poorly documented; the
                 fate and metabolism of toxins in humans is unknown.


                 In every category, the lack of sensitive, specific assays (for research and/or shipboard and
                 dockside testing) and toxin standards were identified as major impediments.

                 The working groups met and proposed recommendations to address each of the impediments.
                 These recommendations, too numerous to list here, are described in detail in the report. They
                 are also embodied in the following goal and objectives (non-prioritized) of the National Plan for
                 Marine Biotoxins and Harmful Algae.


                                                                  E-2










               GOAL: Effective management of fisheries, public health, and ecosystem problems related to
                          marine biotoxins and harmful algae.

               SPECIFIC OBJECTIVES:

               ï¿½   To isolate toxins and their natural derivatives, and characterize their chemical structures and
                   pharmacological action.

               ï¿½   To develop specific detection methods based on the unique chemistry and/or pharmacology
                   of individual toxins.


                   To develop forecasting capabilities for the occurrence and impacts of harmful marine algal
                   blooms.


               ï¿½   To determine the source, fate, and consequences of algal toxins in marine foodwebs and
                   fisheries.

               ï¿½   To develop management and mitigation strategies to minimize impacts of marine bi6toxins
                   and harmful algae.

               ï¿½   To identify and improve access to databases for bloom incidence, toxin occurrence in
                   shellfish, mass mortality events, and epidemiology.

               ï¿½   To develop communication programs that incorporate educational and public health materials,
                   electronic communication, and on-site training.

               ï¿½   TO provide for rapid response to toxic and otherwise harmful marine algal outbreaks.



















                                                              E-3











                                                        PREFACE




              Marine biotoxins and harmful algae represent a significant and expanding threat to human health
              and fisheries resources throughout the United States. This problem takes many forms, ranging
              from massive "red tides" or blooms of cells that discolor the water, to dilute, inconspicuous
              concentrations of cells that are noticed only because of the harm caused by the highly potent
              toxins these cells contain. The impacts of these phenomena include mass mortalities of wild and
              farmed fish and shellfish, human intoxications or even death from contaminated shellfish or fish,
              alterations of marine trophic structure through adverse effects on larvae and other life history
              stages of commercial fisheries species, and death of marine mammals, seabirds, and other
              animals.


              The nature of the problem has changed considerably over the last two decades in the United
              States. Where formerly a few regions were affected in scattered locations, now virtually
              every coastal state is threatened, in many cases over large geographic areas and by more
              than one harmful or toxic algal species. There is a growing. consensus in the scientific
              community that the number of harmful events and the economic costs associated with them have
              increased dramatically over the last several decades in the United States and around the world.
              The reasons for this expansion are the subject of considerable debate. Possible explanations
              include: the eutrophication of coastal waters by human activities, leading to a selection for, and
              proliferation of, harmful algae; increased aquaculture operations which, in other parts of the
              world at least, have been shown to enrich surrounding waters and stimulate algal growth, as well
              as to introduce fisheries resources which simply reveal the presence of previously undetected
              harmful algae; climatic changes; and increased scientific and regulatory scrutiny of coastal waters
              and fisheries products leading to the rapid discovery of toxic events.

              The United States research, monitoring, and regulatory infrastructure is not adequately
              prepared to meet this expanding threat. The present approach is to manage threatened
              fisheries resources using state-run monitoring programs and harvesting restrictions. When
              unexpected outbreaks occur, the response has often been confused, uncoordinated, and slow. This
              approach has, nevertheless, provided a reasonable level of protection to the seafood consumer,
              but illnesses and deaths from marine biotoxins have still occurred, and public confidence in
              seafood safety continues to erode. In addition to these public health concerns, other impacts of
              harmful algae can be significant, including the loss of marketable resources because of fish and
              shellfish mortalities, loss of income for fishermen during outbreaks, or unseen and potentially
              significant effects on marine trophic structure. No single federal agency has assumed a
              leadership role in coordinating and supporting the studies needed to optimize management and
              mitigation strategies. Research funding has always been sporadic and limited.

              In an effort to surmount these problems, a workshop, supported by Saltonstall-Kennedy funds,
              was convened at the NMFS Charleston Laboratory to formulate a National Plan for the
              prediction, control, and mitigation of the effects of harmful algal blooms on marine biota of the


                                                               i










                 United States and to promote the safe consumption of seafoods. Participants were selected to
                 represent all critical scientific disciplines and all geographic regions of continental North
                 America. These individuals were selected from the ranks of academia, the Food and Drug
                 Administration, state public health services, the fishing industry, NOAA/NMFS and the National
                 Sea Grant Program. Attendance was limited to promote close working relationships during the
                 workshop.

                 Prior to the workshop, 12 topics of importance were identified:

                 ï¿½ Toxin pharmacology/epidemiology
                 ï¿½ Toxin analysis/assays/chemistry/standards
                 ï¿½ Bloom biology/ecology
                 ï¿½ Remote sensing
                 ï¿½ Nutrient/pollution effects
                 ï¿½ Taxonomy/genetics/population biology of harmful microalgae
                 ï¿½ Hydrography/physical oceanography
                 ï¿½ Phytoplnkton monitoring
                 ï¿½ Shellfish monitoring
                 ï¿½ Foodweb effects
                 ï¿½ Shellfish depuration/physiology
                 ï¿½ Fish mortalities


                 Workshop participants were assigned topics to review according to their areas of expertise. A
                 group leader was selected for each topic and requested to prepare a position paper with the
                 assistance of others in each group. The position papers, addressing background, current state of
                 knowledge, impediments limiting progress, and prioritized research topics, were distributed to all
                 participants prior to the workshop.

                 At the beginning of the workshop three working groups were formed. The Toxins working group
                 was assigned the first two topics identified above, the Bloom Biology and Ecology working group
                 the next six topics, and the Fisheries and Food Webs working group the last four topics. During
                 individual working group deliberations, modified lists of impediments and recommendations were
                 prepared. These lists were presented to all participants in two plenary sessions for further
                 discussion and modification.


                 Given this procedure and the related nature of the issues discussed by the different working
                 groups, it is not surprising that some issues appear several times in the lists of impediments and
                 recommendations that follow. Rather than arbitrarily removing these common issues from
                 subsequent sections after their first mention, the lists were left unchanged so as to emphasize the
                 cross-disciplinary importance of certain issues.

                 There was considerable discussion on the need to prioritize the recommendations. Attempts to
                 do this in some of the working groups were not successful, as most participants felt that the list
                 of recommendations had already been distilled from a much larger list generated in the position


                                                                ii









              papers, and thus reflected priority issues. Another concern was that it was difficult to establish
              absolute priorities between very different topics (e.g. toxin chemistry issues versus bloom
              dynamics), since such decisions would vary dramatically among individuals or agencies with
              different responsibilities or interests. Accordingly, the recommendations in this National Plan
              are not prioritized. They are grouped by topic, so that agencies developing research or
              monitoring programs can work from the lists that most closely match their purview. An are
              deemed of high (and equal) priority, though it should be recognized that several recommendations
              appear in each of the three topic areas and thus have general applicability across all disciplines.












             1. INTRODUCTION

             in recent years, the incidence of harmful algal blooms (sometimes called "red tides") has
             increased in frequency, severity, and duration, both nationally and globally (Anderson, 1989;
             Smayda, 1990). Tlese episodes are not attributable to a single algal class but rather to a variety
             of physiologically diverse species. Some have long been recognized as problem species, others
             have previously been considered harmless, and still others were unknown to science until their
             initial outbreaks. The causes for this apparent expansion are unknown, but some believe that
             human alteration of the water quality of the coastal zone is an important factor (Smayda, 1990).

             Economic losses in the United States total midlions of dollars per year, and include the cost of
             toxin monitoring programs, closures of harvestable shellfish resources, mortalities of wild and
             farmed fish and shellfish, and the valueof resources that are not exploited or developed because
             of the presence or threat of toxic outbreaks. The United States has managed these threatened
             resources through state-run toxin monitoring programs and harvesting restrictions. Federal
             agencies have provided relatively minor and often unsustained research support for local or
             regional  studies, and state support has been even smaller and more sporadic. Research teams
             have made some progress in developing methodologies for toxin analysis, in understanding the
             structure and pharmacology of certain toxins, in. investigating the physiology of toxin production
             in algae and depuration from shellfish, and in documenting the abundance and distribution of
             certain harmful species during blooms. Despite these efforts, however, we remain woefully
             ignorant of the complex mechanisms underlying the growth and accumulation of individual algal
             species in blooms, the transfer and fate of toxins through the food chain, and perhaps most
             disturbingly, the influence of human activities on these processes. Also lacking are many of the
             tools needed for efficient management of potentially toxic fish or shellfish. In particular,
             sensitive, rapid alternative assay methods are needed for dockside or market-place testing.

             The United States lags far behind many other countries in its approach to the management of
             problems caused by harmful algae and marine biotoxins. Canada, France, Norway, Sweden,
             China, and others have coordinated national research programs that include workshops or
             meetings to exchange results and search for solutions to common problems, sustained funding
             in directions identified as being of high priority, and continual re-evaluation of progress and plans
             for the future. The United States, in contrast, has had only small, fragmented research programs
             carried out by individual investigators, with small budgets that are rarely sustained through time.
             Thus, there is often insufficient communication between U.S. workers and no coordination of
             activities with respect to national priorities.

             The shortfall in our ability to understand and manage these growing problems was reflected in
             fiscal years '91 and '92 priorities for Saltonstall-Kennedy funded research published in the
             Federal Register, many of which related to marine biotoxins. 71is emphasis on biotoxins is but
             one manifestation of the growing awareness that more of our national resources must be focused
             on this topic. During the past several years, other federal agencies have announced specific
             coastal research initiatives, some of which could, and should, include components on marine


                                                               I







                  biotoxins and harmful algae. It is disturbing to recognize that these initiatives are targeting a
                  field that historically has been fragmented, uncoordinated, and poorly funded. Sound input is
                  urgently needed from scientists, industry, and regulatory officials to keep these new research
                  initiatives focused on high priority, productive endeavors. Thus, the primary goal of this
                  workshop was to formulate a National Plan, consisting of a series of recommendations intended
                  to address the major impediments to progress in the management of, and scientific research on,
                  harmful marine algae and associated toxins.

                  The participants formulated the following overall goal and objectives for a National Plan on
                  Harniful Algae and Marine Biotoxins:

                  GOAL: Effective management of fisheries, public health, and ecosystem problems related to
                            marine biotoxins and harmful algae.

                  OBJECTIVES:


                  ï¿½   To isolate toxins and their natural derivatives, and characterize their chemical structures and
                      pharmacological action.

                  ï¿½   To develop specific tests based on the unique chemistry and/or pharmacology of individual
                      toxins.


                  ï¿½   To develop forecasting capabilities for the occurrence and impacts of harmful marine algal
                      blooms.


                  ï¿½   To determine the source, fate, and consequences of algal toxins in marine foodwebs and
                      fisheries.


                  ï¿½   To develop management and mitigation strategies to minimize the impacts of marine
                      biotoxins and harmful algae.

                  ï¿½   To identify and improve access to databases for bloom incidence, toxin occurrence in
                      shellfish, mass mortality events, and epidemiology.

                  ï¿½   To develop communication programs that incorporate educational and public health materials,
                      electronic communication and on-site training.

                  ï¿½   To provide for rapid response to toxic and otherwise harmful marine algal outbreaks.










                                                                  2







               II. THE TOXINS


                  1. General Background

               In the United States, the most significant economic and public health problems related to harmful
               algae are:

               ï¿½  Paralytic shellfish poisoning (PSP), which occurs in all coastal New England states as well
                  as New York and along much of the west coast from Alaska to California. This problem has
                  also extended to offshore areas in the northeast (causative species - the dinoflagellates
                  Alexandriwn tamarense, A.fiundyense, and A. catenella; Anderson et al., 1982; Nishitani and
                  Chew, 1988; Price and Kizer, 1990).

               ï¿½  Neurotoxic shellfish poisoning (NSP) and fish mortalities in the Gulf of Mexico and, more
                  recently, "tending northward to the coast of the Carolinas (causative species - the
                  dinoflagellate Gymnodinlwn breve; Baden et al., 1984; Tester et al., 1991).

               ï¿½  Mortalities of farmed salmonids; in the Pacific Northwest (causative species - the diatoms
                  Chaetoceros convolutus and C. concavicornis and the raphidophyte Heterosigma akashiwo;
                  Homer et al., 1990).

               ï¿½  Recurrent brown tides causing mass mortalities of mussel populations in Rhode Island,
                  massive recruitment failure of scallops, and reduction of eelgrass beds around Long Island
                  (causative species - the previously unknown chrysophyte, Aureococcus anophagefferens;
                  Sieburth, et al., 1988).

               ï¿½  Ciguatera fish poisoning (CFP), a malady associated with dinoflagellate toxins accumulated
                  in tropical fish flesh, occurring in virtually all sub-tropical to tropical U.S. waters (Florida,
                  Hawaii, Guam, U.S. Virgin Islands, Puerto Rico, and many Pacific Territories; Ragelis, 1984;
                  major causative species Gambierdiscus toxicus, Prorocentrwn spp., Ostreopsis spp., Coolia
                  monods, Thecadinwn sp., and Amphidiniwn carterae; Juranovic and Park, 1991).

               ï¿½  Amnesic shellfish poisoning (ASP) which occurred first in southeastern Canada in 1987, but
                  has been a problem for the U.S. Pacific coast states over the past two years (causative
                  species - the diatoms Pseudonitzschid pungens forma mulliseries and Pseudonitzschia
                  australis; Garrison et al., 1992; Buck et al., 1992; Fritz et al., 1992; Wood and Shapiro,
                  1992). This sometimes fatal illness is so named because one of its most severe symptoms
                  is the permanent loss of short-term memory. The ASP toxin, domoic acid, has been detected
                  in shellfish from both the West and East Coasts of the United States, and toxic P. pungens
                  f. multiseries cells have been isolated from Gulf of Mexico waters, though no toxin has yet
                  been detected in the field. 'Mus, the threat to U.S. shellfish consumers from this dangerous
                  alga covers a broad geographic area. The name "ASP" understates the severity of this
                  problem, as it is now known that domoic acid also accumulates in fish and in crab viscera
                  along the west coast of the United States, where the impact of this toxin on non@molluscan
                  fisheries may well exceed the loss to molluscan fisheries (e.g., razor clam).



                                                                3







                  Another serious threat is diarrhetic shellfish poisoning (DSP) which some consider the most
                  serious and globally widespread phytoplankton-related seafood illness. The first confirmed
                  incidence of DSP in North America occurred in 1990 when these toxins were detected in shellfish
                  from the southern coast of Nova Scotia following numerous human illnesses Quilliarn et al.,
                  submitted ms.). Another DSP outbreak in Canada occurred in 1992 (Wright, pers. comm.). DSP-
                  producing species of phytoplankton occur throughout all temperate coastal waters of the United
                  States, and thus present a potential problem for the future, though no outbreaks of DSP have yet
                  been confirrned.


                      2. Toxicology and Pharmacology

                         2.1 Background

                  Naturally-occurring toxins responsible for the intoxication syndromes associated with seafood are
                  as diverse as are the algae that produce them (Table 1). Man is exposed principally

                                                                TABLE 1


                                             TOXIN-DERIVED HUMAN TOXICOSES



                                TOXIN FAMILY             SYNDROME' SOLUBILITY                  ACTION ON
                              (Number of Toxins)

                                 Brevetoxin (10)              NSP                Fat           Nerve, Muscle,
                                                                                                Lung, Brain
                             Ciguatoxin /Maitotoxin           CFP             Fat/Water        Nerve, Muscle,
                                    (multiple)                                                  Heart, Brain
                                Dornoic Acid (11)             ASP               Water              Brain
                                Okadaic Acid (3)              DSP                Fat              Enzymes
                                  Saxitoxin (18)              PSP               Water           Nerve, Brain

                              NSP= Neurotoxic Shellfish Poisoning, ASP       Amnesic shellfish Poisoning,
                             DSP = Diarrhetic Shellfish Poisoning, PSP = Paralytic Shellfish Poisoning,
                             CFP = Ciguatera Fish Poisoning

                  by consumption of contaminated seafood products, although one type, of toxin (brevetoxin),
                  because of aerosol formation due to wave action, also causes respiratory asthma-like symptoms
                  during blooms of the toxigenic organism. In all cases, the diseases are caused by specific
                  interaction of the toxins with tissues and organs responsible for carrying out vital cellular
                  functions. By modifying these functions in deleterious ways, the toxins disrupt nerve electrical
                  conduction, uncouple communication between nerve and muscle, and prevent critical
                  physiological processes from occurring. Most of the toxins accomplish this by binding to
                  specific receptors, or docking sites, on the tissue or organ leading to critical changes in
                  intracellular concentration of ions such as sodium, calcium, and potassium. Some of the cellular
                  changes lead to permanent effects in the exposed cells.



                                                                     4







             Seafood toxins bind with high affinity to specific receptor sites, often with binding constants in
             the  10-1 to 10-12 M range.    Most binding is reversible, but dissociation times may be quite
             prolonged. Except for identification of the general category of toxin receptors in living
             organisms, virtually nothing is known about the chemical interaction of the toxins with their
             specific binding sites.

             Many of the toxin classes are     not single chemical entities, but instead represent families of
             compounds of similar chemical     structure (Table 1). Each toxin derivative of the same parent
             compound is slightly altered in chemistry. This leads to wide-ranging variability in toxicity of
             the individual modified toxins.


             Acute single-dose lethality of seafood toxins has been extensively studied in the laboratory
             (Shimizu, 1987). However, chronic and/or repeated exposure to marine seafood toxins, which
             is a more realistic phenomenon, has not been adequately examined. There is a serious lack of
             knowledge as to how the toxins are distributed throughout the body and eliminated. Other
             important questions include how long the toxins circulate before elimination, and how they are
             metabolized by living organisms. These knowledge gaps prevent researchers from devising
             antidotes or effective treatments which may alleviate or lessen the symptoms. Therapeutic
             intervention is primarily limited to symptomatic treatment and life support if necessary.

             Similarly, statistical data collection on human exposure, intoxication duration, and number of
             incidences are limited and incomplete. Many cases of, intoxication are not reported, or are
             reported inadequately based on hear-say evidence with little documentation.

             Since 1978, illnesses in the U.S. due to natural algal toxins have included CFP, PSP, NSP and
             ASP. No incidents of DSP have yet been verified in this country. Although records are
             incomplete because reporting to the Centers for Disease Control (CDC) is voluntary, evidence
             indicates that ciguatera was responsible for about half of all seafood intoxications between 1978
             and 1987 (Ahmed, 1991). A growing body of evidence indicates that incidents of ASP are on
             the increase (Buck et al., 1992; Garrison et al., 1992; Villac et al., in press; Homer and Postel,
             in press), and that DSP may shortly make its d6but in the United States. Certain of the toxicoses,
             like the toxigenic organisms, are focused geographically and result from consumption of
             particular species. However, with the increase in interstate and international transport of seafood,
             as well as international travel by seafood consumers, there are virtually no human populations
             that are free from risk.


                  ASP: Amnesic shellfish poisoning can be a life-threatening syndrome. It is characterized
             by both gastrointestinal and neurological disorders (Bates et al., 1989). Gastroenteritis usually
             develops within 24 hours of the consumption of toxic shellfish; symptoms include nausea,
             vomiting, abdominal cramps, and diarrhea. In severe cases, neurological symptoms also appear,
             usually within 48 hours of toxic shellfish consumption. These symptoms include dizziness,
             headache, seizures, disorientation, short-term memory loss, respiratory difficulty, and coma. In
             1987, four victims died after consuming toxic mussels from Prince Edward Island, Canada. Since
             that time, Canadian authorities have monitored both the water column for the presence of the
             causative diatom, and shellfish for the presence of the toxin, domoic acid. Shellfish beds are
             closed to harvesting when the domoic acid concentration reaches 20 pg/g shellfish meat. Fish


                                                              5








                  and crab viscera can also contain domoic acid, so the risk to human consumers and animals in
                  the marine food chain is more significant than previously believed.

                  Ciguatera: Ciguaterafish poisoning produces gastrointestinal, neurological, and cardiovascular
                  symptoms. Generally, diarrhea, vomiting, and abdominal pain occur initially, followed by
                  neurological dysfunction including reversal of temperature sensation, muscular aches, dizziness,
                  anxiety, sweating, and a numbness and tingling of the mouth and digits. Paralysis and death have
                  been documented, but symptoms are usually less severe although debilitating (Miller, 1991).
                  Recovery time is variable, and may take weeks, months, or years. Rapid treatment (within 24
                  hours) with manitol is reported to relieve some symptoms. There is no antidote, supportive
                  therapy is the rule, and survivors recover. Absolute prevention of intoxication depends upon
                  complete abstinence from eating any tropical reef fish, since there is currently no easy way to
                  measure routinely ciguatoxin or maitotoxin in any seafood product prior to consumption.

                  DSP: Diarrhetic shellfish poisoning produces gastrointestinal symptoms, usually beginning
                  within 30 min to a few hours after consumption of toxic shellfish (Yasumoto and Murato, 1990).
                  The illness, which is is not fatal, is characterized by incapacitating diarrhea, nausea, vomiting,
                  abdominal cramps, and chills. Recovery occurs within three days, with or without medical
                  treatment.


                  NSP: Neurotoxic shellftsh poisoning produces an intoxication syndrome nearly identical to that
                  of ciguatera. In this case, gastrointestinal and neurological symptoms predominate. As noted
                  above, formation of toxic aerosols by wave action can produce respiratory asthma-like symptoms.
                  No deaths have been reported and the syndrome is less severe than ciguatera, but nevertheless
                  debilitating. Unlike ciguatera, recovery is generally complete in a few days. Monitoring
                  programs (based on G. breve cell counts) generally suffice for preventing human intoxication,
                  except when officials are caught off-guard in previously unaffected areas.

                  PSP: Paralytic shellftsh poisoning, like ASP, is a life threatening syndrome. Symptoms are
                  purely neurological and their onset is rapid. Duration of effects is a few days in non-lethal
                  cases. Symptoms include tingling, numbness, and burning of the perioral region, ataxia,
                  giddiness, drowsiness, fever, rash, and staggering. The most severe cases result in respiratory
                  arrest within 24 hours of consumption of the toxic shellfish. There is no antidote, supportive
                  therapy is the rule and survivors recover. PSP is prevented by large-scale proactive monitoring
                  programs (assessing toxin levels in mussels, oysters, scallops, clams) and rapid closures to harvest
                  of suspect or demonstrated toxic areas.

                       2.2. Impediments and Recommendations

                  The Toxins working group identified three major impediments to progress in the area of toxin
                  pharmacology and toxicology, and recommended solutions to these impediments. Priority order
                  was not assigned to the impediments.

                  IMPEDIMENT: Reference toxin is difficult to obtain, is not always reproducible, and is
                  generally costly. This impedes development of methods for detection, prevents detailed studies
                  of physiology, and inhibits development of molecular pharmacology to explain toxin interaction
                  at the receptor level.

                                                                  6







             RECOMMENDATION: Establish reference toxin supplies for the five major classes of
             toxins.

             This can be accomplished by isolation and purification of toxin from fish and shellfish, from
             mass cultures of the toxic phytoplankton, or synthesis of less accessible toxins. Production of
             radiolabeled toxin first requires adequate supplies of purified standards. Three levels of standards
             are required: Pharmacological Standards, Analytical Standards, and Certified Standards.

             Toxic marine dinoflagellates are some of the most difficult algae to grow in mass culture.
             Facilities required are extensive and sophisticated, and careful control of nutrient levels, pH,
             temperature, lighting, security, and cleanliness is necessary. Likewise, the facilities necessary
             to extract and purify multi-milligram quantities of these highly potent materials are extensive, and
             require special instrumentation for separation, isolation, detection and storage. At many such
             facilities, detailed safety plans are implemented. Although this is an objective which should be
             achieved as rapidly as possible, it must be recognized that a continuing financial commitment is
             necessary to implement the production of consistent, reliable standards. With the exception of
             ciguatera-related toxins, facilities expansion to produce the desired quantities of all levels of
             standards is a logistical possibility within 2-3 years. The ciguatera toxins will require extensive
             research prior to standards availability (see toxin standards section).

             A major problem with available biotoxin supplies is their distribution to monitoring agencies and
             the research community. A well-defined distribution plan for biotoxin standards isolated or
             synthesized with the assistance of federal funds needs to be prepared. It is an overwhelming
             consensus that toxins should be made available at a minimal cost, for example, as is done by the
             National Hormone and Pituitary Program.

             IMPEDIMENT: The annual incidence of seafood toxin poisoning is poorly documented.

             Without knowledge of the actual magnitude of the problem, little can be done to evaluate
             remedial measures aimed at reducing incidence. Effects of episodic and chronic exposure have
             been totally neglected. It is also apparent that part of the reporting problem is due to lack of
             toxic syndrome recognition.

             RECOMMENDAT71ON: Develop a database in collaboration with the CDC for marine
             seafood intoxication. Explore development of better reporting tools, including mandatory
             reporting. Incorporate episodic or chronic exposures. Educate physicians, public health
             officials, and consumers in issues of seafood poisoning.

             Incidences of seafood intoxication are thought to far exceed the actual reporting. Valuable
             information regarding seafood intoxication does not reach the proper reporting officials for many
             reasons. Initial symptoms often are gastrointestinal in nature and are misdiagnosed as the "flu."
             Only later, after intensification of symptoms or a significant number of individual events, are
             alternative diagnoses considered. Early identification of the toxicologic syndrome is necessary
             to permit effective therapeutic intervention. Once recognized, proper and prompt reporting alerts
             official agencies to implement regulatory directives. If the syndromes are recognized, and the
             reporting is' mandatory, the data will be comprehensively recovered. This is a long-term
             objective, and has appended to it a continuing effort to.collect and analyze data, to educate, and

                                                               7







                 to develop the program further.       Initial funds (1-2 yrs) should be provided to trained
                 epidemiologists and biostatisticians for examining the status of the situation and providing
                 additional recommendations for full implementation of a program. Ideally, this should be
                 undertaken by an epidemiologist or group of epidemiologists with interest and experience in
                 marine toxin poisoning. Full implementation of this program would take 5-10 years.

                 IMPEDIMENT: Poorly-defined aspects of molecular pharmacology prevent development of
                 therapeutic agents and appropriate receptor-based assays for marine toxins.

                 RECOMMENDATION: Identify primary tissues of toxicologic action in animals and man,
                 and develop in vitro models that reflect the primary toxicologic action. Identify the
                 molecular      characteristics    of specific       receptors    for     saxitoxin/tetrodotoxin,
                 brevetoxin/ciguatoxin, maitotoxin, okadaic acid, and domoic acid. Explore structural
                 modification of toxins in relation to toxicity and binding affinity.

                 The molecular mechanism of intoxication is not known in detail for any marine toxin. This is
                 in large part due to the fact that we do not know the primary targets for the biotoxin in the
                 human (or animal) body. It is critical to define the primary relevant site of action for marine
                 biotoxins, before funding substantial amounts of research on nonrelevant tissues.

                 Laboratory studies which employ radiolabeled toxins are capable of defining specific binding or
                 recognition sites on a molecular level. Detailed knowledge of the toxin receptors is essential
                 for understanding why "toxins" are toxic. Elucidation of the mechanism of action provides
                 information essential for the development of receptor-based methods for detection, and practical
                 methods for treatment of intoxication. Receptor-derived assays correlate well with toxicity, and
                 may be enhanced using molecular recombinant technology, thereby reducing the need for animal-
                 based toxicological assays. With our current capabilities and available molecular probes,
                 brevetoxin research will continue (complete in 2-3 years), and pharmacologic probe synthesis and
                 investigation initiated for saxitoxin, okadaic acid, and domoic acid (3-6 yrs). Maitotoxin and
                 ciguatoxin(s) will require more structural information prior to direct analysis (> 7 yrs).

                     3. Analysis, Standards, Chemistry

                        3.1. Background

                 The chemical structures of ASP, DSP (okadaic acid family), NSP and PSP toxins are known
                 (Baden, 1984; Shimizu, 1984; Yasumoto et al., 1984; Shimizu et al., 1986; Wright et al., 1989;
                 Hall and Strichartz, 1990). Structures of the toxins responsible for ciguatera are not yet known,
                 except for ciguatoxin, in fish from the Tahitian region (Murata et al., 1990). In addition, the
                 structure and role of maltotoxin (MTX) is only partially understood (Murata et al., 1992), as is
                 its role in ciguatera fish poisoning. Determination of the structures of newly-recognized marine
                 biotoxins requires the dedication of substantial fiscal resources, instrumentation, and personnel
                 time. Currently, the only certified standard(s) available is for domoic acid (Institute for Marine
                 Biosciences, National Research Council of Canada, Halifax, NS, Canada). Reference material
                 is also available for domoic acid. Moreover, suites of standards for the differeni naturally-
                 occuffing toxins are necessary.


                                                                 8







             Biological tests using mice or rats are available for all listed toxins, but the bioassay for ASP
             toxins lacks sufficient sensitivity Quilliam et al., 1989). Other biological tests that do not use
             animals have yet to be subjected to collaborative review. Accepted chemical analytical methods
             have been developed only for the ASP toxin, domoic acid Quilliam, et al., 1991; Pocklington,
             et al., 1990). Chemical methods exist for the other toxins but have not gone through appropriate
             collaborative review (e.g., for PSP) or need further refinement (CTX, DSP, NSP).

             In recent months, the detection of known toxins in unexpected vectors (e.g., PSP toxins in crab
             viscera, ASP toxins in fish and crab viscera) has accentuated deficiencies in assay procedures.
             The extension of certain assay procedures to "new" tissues can sometimes be done routinely when
             the organisms are related, but new procedures need to be developed when organisms not covered
             by the original method are investigated. Substitution of marine marnmal liver for clarn tissue in
             PSP testing, for example, can lead to erroneous conclusions.


                    3.2. Impediments and Recommendations

             The Toxins working group identified three major impediments to progress associated with the
             development of standards and assays and provided recommended solutions to these non-
             prioritized impediments.

             IMPEDIMENT: The chemical complexity of marine biotoxins impedes the development of a
             viable seafood safety program and successful flsheries resource management.

             RECOMMENDATION: Determine the structure, chemical properties, and pharmacological
             behavior of marine toxins. Develop methods for extraction and purification of toxins from
             natural sources, their synthesis, and preparation of toxin derivatives.

             71be determination of the structure, properties, and behavior of marine toxins is an essential first
             step for the development of new and improved methods for the detection of toxins, for mitigating
             their presence and,effects in seafoods, and for a successful fisheries resource management
             program. This expanded knowledge will augment the characterization of new seafood toxins as
             they arise. The structural characterization of marine toxins has challenged researchers for many
             years due to lack of appropriate expertise, resources, and specialized equipment and techniques
             not commonly available, such as nuclear magnetic resonance (NMR) spectrometers and "soft"
             ionization mass spectrometers.

             Mass culture of toxic algae, chemical synthesis, chemical modification, and derivatization
             methods are necessary for the economical production of toxin standards and related compounds,
             and are vital for the preparation of easily-detected derivatives for use in monitoring programs and
             toxicological studies. Toxicological and pharmacological studies are necessary for the
             development of antidotes or other medical intervention techniques. These activities might require
             3-5 years for known toxins, and 5-7 years for unknown or poorly understood toxins.

             IMPEDIMENT: Every study related to marine biotoxins is compromised by a lack of toxin
             standards. It is impossible to develop methods for the detection and quantiflcation of marine
             toxins without the availability of deflned marine toxin standards.

                                                              9







                  RECOMMENDATION: Isolate or synthesize, and characterize sufficient quantities of
                  purified toxin(s) for the development of appropriate standards. Establish a toxin standard
                  development, maintenance, and distribution system, so that these toxins are equally
                  accessible and available to all qualified research groups.

                  Biological, toxicological, and chemical studies require submilligrarn to milligram quantities of
                  rigorously characterized marine toxins. This work will require the collaboration of biologists to
                  produce adequate quantities of source materials, and chemists to isolate, purify, and characterize
                  the toxins.   Once prepared, either by biological or synthetic means, these toxin standards must
                  be readily and reliably available to research and monitoring programs so that public health is
                  adequately protected and fisheries resources are effectively managed. 17hese standards should
                  have international acceptance.

                  IMPEDIMENT: Current assay methods for marine biotoxins are, inadequate for most
                  monitoring and research purposes. Most monitoring programs utilize whole animal assays
                  which lack sensitivity and are becoming increasingly unacceptable.

                  RECOMMENDATION: Develop rapid and cost-effective methods for detecting and
                  quantifying marine toxins that are internationally acceptable.

                  The seafood industry is a global activity that requires coordination of safety regulations between
                  trading nations. To avoid conflicts, the development of marine toxin action levels and methods
                  must be evolved in close cooperation with other international agencies. A particularly important
                  challenge is the development of rapid field methods, such as enzyme or irnmunoassay-based ldts.
                  Sensitivity of all methods must relate to human toxicity and to levels present in contaminated
                  seafoods. The methods developed must be subjected to verification through formal, collaborative
                  studies.





























                                                                    10







            III. BLOOM BIOLOGY AND ECOLOGY


                  1. Bloom Dynamics

                     1.1. Background

            The impacts from harmful or toxic blooms are necessarily linked to the population size and
            distribution of the causative algae. Efforts to manage fisheries resources affected by algal blooms
            or to assess the possible impacts of anthropogenic influences on harmful species requires an
            understanding of bloom biology and ecology.

            The growth and accumulation of individual harmful algal species in a             mixed planktonic
            assemblage are, however, exceedingly complex processes involving an array of chemical,
            physical, and biological interactions. Blooms can occur over wide geographic areas and may
            involve long-distance transport to affected resources. Harmful blooms can also occur on the
            ocean bottom, caused by either microscopic or macroscopic algal species. Macroalgal blooms
            need not produce toxins to be hamful. They can dominate planktonic or benthic communities,
            changing food web structure and altering habitats for many marine organisms.

            Our level of knowledge about each of the many harmful algal species varies significantly, and
            even the best-studied remain poorly characterized with respect to bloom or population dynamics.
            Resolution of various rate processes integral to the population dynamics (e.g., input and losses
            due to growth, grazing, encystment, excystment, and physical            'advection) has not been
            accomplished, but is fundamental to the long-term management of fisheries resources or marine
            habitats affected by harmful algae. Many of the processes are difficult to quantify in the field
            because harmful species are often only a small fraction of the biomass in natural samples. The
            end result is that there are no predictive models of population development,, transport,, and
            toxin accumulation for any of the major harmftil algal species in the United States.

            Within the past two decades, the incidence of toxic blooms caused by formerly undetected taxa
            (the so-called "hidden flora") has increased (Anderson, 1989; Smayda, 1990). The basic biology
            and environmental triggers for toxic activity of these cryptic species have not been characterized.
            U.S. coastal waters are generally becoming nutrient enriched, often because of human influences.
            The impact of increased nutrients on harmful algal bloom events remains uncertain, however, and
            the relative importance of natural variance vs andiropogenic influences on blooms is not known
            (Smayda, 1990). To further confuse the issue, global changes and trends in seveml physical and
            chemical parameters, such as temperature and UV radiation, as well as nutrient enrichment, may
            also affect harmful algal blooms.

            The long-anticipated potential of remote sensing is becoming a reality in the study of harmful
            bloom dynamics. Near real-time sea surface temperatures have been used successfully to identify
            oceanic features and water masses associated with blooms of two hannful species in two different
            hydrographic regimes (Keafer & Anderson, in press; Tester et al., 1991). This approach needs
            further refinement and should be extended to other species and regions of the United States.






                 There is a serious deficiency in our understanding of the physiology and genetics of toxin
                 production. Potentially harmful algae exhibit genetic variability to the extent that toxic and non-
                 toxic strains occur within individual species, and toxic species exhibit a range of inherent
                 potencies.     These differences in toxin composition and content are genetically and
                 environmentally regulated, and increase the difficulty in identifying and evaluating the harmful
                 effects of these algae. Development of molecular probes and other techniques for genetic
                 characterization would aid in the identification and separation of harmful algae present in mixed
                 natural populations.

                           1.2. Impediments and Recommendations

                 The Bloom Biology and Ecology working group identified five major impediments to progress
                 in the'area of algal population. dynamics, biology, and ecology, and one impediment in the area
                 of phytoplankton monitoring. The group recommended solutions to these impediments.

                 IMPEDIMENT: Adequate-documentation of harmful algal events is &fficult because of the
                 lack of rapid, species-specific methodsfor counting and separating cellsfrom natural samples.

                 RECOMMENDATION: Support cooperative development of molecular probes (nucleic
                 acid and, antibody-based) and other techniques for genetic characterization. Provide access
                 to appropriate facilities and equipment; disseminate technology and probes.

                 Harmful,and benign organisms are currently difficult to distinguish in a timely fashion. This
                 restricts'identification and separation of harmful algae for their rapid quantification and analysis
                 within multi-species planktonic assemblages. Nucleic acid and antibody probes, which target
                 different cellular components, offer high flexibility and specificity in their design and application.
                 Some laboratories which have the appropriate skills, expertise, and equipment can provide
                 training and support for. others that have the need for probes but lack these resources. Equipment
                 should be made available for the analysis of field and laboratory samples, perhaps through a
                 dedicated facility with a flow cytometer and equipment for nucleic acid and protein analysis.
                 Probes are in an early stage of development for several harmful species, but for most species,
                 there is no sequence information or other biochemical characterization that can be used to design
                 specific probes. Knowledge about one species can frequently be applied to closely related species,
                 greatly accelerating the rate at which unstudied species can be characterized. With immediate
                 support, species- specific probes for several harmful algae could be available within one to two
                 years. A battery of probes against many harmful species could follow within 5 years. Once the
                 genes involved in toxin production are identified and characterized, probes can be developed to
                 identify only toxic species. The use of such probes in quantifying target cells requires additional
                 studies of the physiological variability of their molecular targets under different environmental
                 conditions.


                 IMPEDIMENT: Population dynamics, including the rate processes required in predictive
                 models of harmful blooms, cannot be adequately described or predicted, although this
                 information is of fundamental importance to effective resource managemenL




                                                                   12







            RECOMMENDATION: Determine biological rate processes and initiate studies of coastal
            hydrography and water circulation for development of physically/biologically coupled
            models at temporal and spatial scales appropriate to harmful algal blooms.

            Despite the fundamental importance of predictive models for harmful algal blooms in different
            regions, no such models exist for U.S. problem species. Knowledge of the rate processes that
            determine the net accumulation of cells and physical models of  the regional hydrographic features
            that influence the initiation, distribution and maintenance of blooms are both indispensable to
            such models.


            Information on bloom dynamics can be gained through laboratory and field studies that define
            nutrient uptake kinetics, growth rates, loss terms, and life cycle dynamics. While field conditions
            such as circulation, meteorology, and water chemistry have long been recognized as critical
            elements in blooms of some toxic species, neither the initial boundary conditions, nor the
            hydrogr aphic regimes within which harmful blooms occur are clearly understood. Regional
            multi-disciplinary field efforts, adequate to characterize the physical circulation models, are
            needed. Ideally, these would be 3-5 year programs. The ultimate goal is to couple population
            dynamics with physical -circulation models for a given hydrographic regime, and to refine the
            physically/biologically coupled models using field bloom observations and toxicity patterns.
            Laboratory studies could be accomplished within about 2 years per species. Field studies can be
            greatly facilitated by timely accessibility to archived and in situ environmental information.

            IMPEDIMENT: Competitive outcomes in species selection and succession cannot be predicted,
            nor can the relative effects of natural vs. anthropogenic factors be resolved.

            RECOMMENDATION: Undertake experimental studies on factors regulating selection and
            succession, emphasizing grazing, nutrients and related anthropogenic variables, and
            allelopathic effects of toxins.

            Prediction of harmful species occurrences and evaluation of potential stimulation by
            anthropogenic influences are essential for effective resource management. Ile few available
            long-term data sets strongly suggest a link between nutrient enrichment and increasing
            occurrences of known harmful species as well as formerly undetected taxa. ("hidden flora").

            Prediction of the outcomes of competitive interactions between harmful algae and other food web
            components depends upon understanding the processes regulating growth, toxicity, and
            encystment of individual harmful species. Laboratory experiments (2-3 years) can be used to
            examine growth across gradients of nutrients (i.e., absolute concentrations and variable supply
            ratios), temperature, salinity, light, mixing, and grazing by appropriate predators. Given this
            knowledge, experiments can be expanded to include natural communities (e.g., in mesocosms,
            field enclosures; 5 years) in order to examine competition, grazing, allelochernical effects, and
            other influences on selection and succession of harmful algae. These field data can be used to
            estimate rate constants for accumulation and loss terms which, in turn, would enable construction
            Pf mathematical models needed to assess mitigation strategies under variable environmental
            conditions.




                                                             13







                  Understanding the influence of anthropogenic effects will require analysis of data bases for
                  phytoplankton communities and tractable anthropogenic variables such as inputs of nutrients and
                  other pollutants. Initiation or expansion of long-term monitoring programs of at least 10-years
                  duration must include both episodic events and nutrient time-series studies. Short-term and long-
                  term correlations between pollutant inputs and abundances of harmful algal species, together with
                  information from the autecological studies, will provide a basis for mesocosm-scale experiments.
                  These experiments (3-5 years duration) are needed to test potential mitigation strategies and
                  strengthen interpretations about the influences of anthropogenic variables on bloom species
                  selection and succession.


                  IMPEDIMENT: There is insufficient knowledge of the physiology of algal growth and toxin
                  production in response to environmenid variables.

                  RECOMMENDATION: Conduct experimental studies of organism physiology, emphasizing
                  environmental tolerances and factors which influence growth and toxin production. Expand
                  culture collections to include broad geographical representation of all potentially harmful
                  species; include multiple clones from single populations.

                  Tolerance ranges and optima for growth and toxin production in response to environmental
                  variables such as salinity, temperature, and light must be determined for multiple toxic and non-
                  toxic (if available) clones of each species in batch culture. In addition, classical steady-state
                  analyses of nutrient requirements and uptake rates and toxin physiology (including content and
                  composition) of each species are necessary. This work will depend upon a supply of appropriate
                  isolates, our ability to manipulate them in culture, and the availability of sensitive and reliable
                  methods of toxin analysis. Physiological experiments can be carried out in tandem with studies
                  of tolerance ranges and optima once the basic individual growth requirements are determined,,
                  and will take 3 yrs to complete for each species.

                  Individual clones of a single species exhibit marked variation in numerous characteristics,
                  including growth and toxin production (Maranda. et al., 1985; Bomber, et al., 1989; Cembella et
                  al., 1987; Hayhome et al., 1989), and thus may not be representative of local or regional
                  populations. A few laboratories in the United States have initiated "syndrome-based" culture
                  collections of harmful marine microalgae. Presently, isolates housed in these collections do not
                  adequately represent the full range of variants characteristic of each harmful species. It is
                  essential that new clones be established from throughout the geographical range of each harmful
                  species. Establishment of clones should be accompanied by basic screening programs in order
                  to select ideal clones for physiological and toxicological studies.

                  Production of toxins in quantities sufficient for their purification and characterization requires
                  identification and culture of "high performance" clones and knowledge of their growth
                  requirements. Such collections could be established within a 3-year period. Completion of basic
                  screenings may extend each project into a fourth year.

                  There are anecdotal and circumstantial accounts of bacterial involvement in toxin production by
                  harmful algal species, but only one set of published data demonstrates bacterial synthesis of PSP
                  toxins (Kodama, 1990). The existence of toxigenic bacteria and their association with harmful
                  algal species must be investigated. This work will rely on the isolation of bacteria and the

                                                                  14






            development of techniques that optimize our ability to detect toxin production. Verification of
            toxigenic bacteria will take about one year for each species of harmful algae once methods are
            accepted for unequivocally demonstrating the presence or absence of the bacteria.

                2. Phytoplankton Monitoring

                   2.1. Background

            Testing shellfish and other seafood for possible toxins is expensive and time-consuming. Further,
            current seafood monitoring efforts are often limited by cost and geographic area covered and may
            not even test the food product most affected by a particular toxin. An easier and possibly more
            effective approach involves regular, routine sampling and analysis of phytoplankton samples,
            especially in areas where aquaculture and/or recreational harvesting are common. If potentially
            toxic phytoplankton species are found, then more expensive seafood testing must be done.

            Routine phytoplankton monitoring would provide long-term data on the occurrence of harmful
            algal species and foster the development of testable hypotheses and insights into the status and
            trends in harmful algal bloom events. Retrospective analyses of the few existing historical data
            sets and initiation of time-series will allow assessment of therole of improved monitoring
            programs and strategies. This information will also promote development of badly needed
            mitigation methods.

                   2.2. Impediments and Recommendations

            IMPEDIMENT: Coastal environmental programs are inadequate for bloom detection,
            monitoying and mitigation of bloom effects.

            RECOMMENDATION: Identify regional expertise and facilities. Establish species-specific
            monitoring programs on a regional scale, using shipboard techniques and remote sensing
            where appropriate. Identify sentinel species appropriate for each specific toxin and habitat
            type. Organize regional response teams and logistical support for unexpected events, and
            reporting centers to accommodate rapid response. Develop practical response protocols for
            protecting aquaculture sites on a regional and/or species-specific basis. Coordinate and
            develop national and regional training programs (e.g., sampling and identification methods).
            Develop and disseminate adequate reference materials.

            Adequate phytoplankton monitoring programs can serve as early warning systems to moderate
            the effects of blooms on public health, aquaculture, and fisheries. Response teams, organized by
            region using existing expertise and maintained as part of a national program, should augment
            species-specific monitoring programs in areas of recurring bloom events. In-water monitoring
            and remote sensing provide the early warning systems needed by the aquaculture industry and
            government officials. Further, long-term data sets are needed for trend analyses.           These
            recommendations are a high priority and must be implemented through federal/state/
            academic/private industry partnerships.

            A network of sentinel sites might be composed of local residents and user groups who are often
            the first to recognize a bloom event and notify local government agencies. Other sentinel sites

                                                           15







                  could be located at coastal aquaculture facilities. Government and/or industry personnel must
                  be able to sample and quickly identify the causative organism and determine whether it is a
                  known or potentially toxic species. Samples must be sent to taxonomic experts for verification.
                  Technical training will provide the expertise needed for early warning systems and local response.
                  Training should be structured at several levels.

                  These recommendations should be implemented immediately and continue indefinitely, although
                  possibly on a somewhat reduced level after 5 years, depending on trend analysis and local needs.
                  The Canadian domoic acid experience has shown that phytoplankton monitoring can be an
                  effective component in a program to protect seafood consumers Erorn marine biotoxins.

                  IMPEDIMENT: The causes and effects of harmful blooms of benthic and planktonic
                  macroalgal species are poorly understood.

                  RECOMMENDATION: Evaluate the manner in which macroalgal species composition can
                  be influenced by nutrient enrichment, coastal erosion, and other human activities.
                  Determine the effects on habitats and food-chain structure that are associated with
                  macroalgal blooms.

                  Much of the focus in this program is on microscopic algal species which bloom in surface waters,
                  but harmful blooms of macroalgae also occur. These can cause harm by altering benthic habitats
                  through the displacement of indigenous species, and by changing food-chain structure and
                  dynamics. One manifestation of coastal nutrient enrichment is the enhancement of benthic (and,
                  on occasion, planktonic) macroalgal abundance, with certain opportunistic species often
                  dominating. Not only will studies of benthic algal species succession and dominance be
                  necessary for effective management of coastal resources, but the changing distribution and
                  abundance of these species through time and space may provide strong evidence of the extent
                  of human impacts on algal populations in general.





















                                                                  16








             IV. FISHERIES AND FOOD WEBS


                 1. General Background

             As biotoxins move up through marine food webs, they can have a broad spectrum of effects on
             marine animals in inshore, offshore, pelagic, and benthic habitats (Table 2). The scope of these
             effects, resulting from both chronic and acute exposure to the toxins, has become more evident
             in recent years (Anderson and White, 1989; White, 1980, 1988; White, in press; White et al.,
             1989). A wide variety of animals can accumulate biotoxins and act as intermediate vectors to
             consumers at higher trophic levels. Certain groups of animals, as direct consumers of microalgae,
             have received primary attention with regard to specific biotoxins. The best-known examples are
             filter-feeding bivalve molluscs as vectors for PSP, NSP, DSP, and ASP (Shurnway, 1990).
             Phycotoxins: are, however, increasingly being detected in a wide range of marine animals, such
             as gastropod molluscs, zooplankton, planktivorous fish, benthic: crustaceans, sea birds and marine
             mammals (Quayle, 1969; Halstead, 1978; White, 1981b; Smayda, 1992).

             Marine fish and shellfish kills caused by harmful algae may have significant economic impacts
             on coastal communities through lost recreational and commercial fishing revenues and adverse
             aesthetic effects on tourism (e.g., fish kills in Florida and the southeastern United States) and
             decimation of bay scallop stocks and reduction of eelgrass nursery habitat by brown tides in New
             York; Cosper et al., 1987). Harmful algae also may have direct (non-food chain) and
             catastrophic economic effects on finfish aquaculture. There is no uniform recording or reporting
             of fish kills, but the frequency of these events may be increasing.

             Coastal waters in the United States harbor a number of harmful phytoplankton species that could
             cause, or already may have caused, massive fish and shellfish mortality, judging from recent
             events in other parts of the world. For. example, Chatonella andqua, various silico-flagellates,
             and Prymnesiwn spp. are present here but have not been documented to cause the fish and
             shellfish kills and other mortality events seen elsewhere in the world. Other toxic species remain
             to be identified, such as an unusual dinoflagellate species responsible for a number of fish kills
             in North Carolina (Burkholder et al., 1992). It is possible, even likely, that this dinoflagellate
             has caused fish kills in all of the mid-Atlantic states for decades or more.


             It is known that biotoxin conversions (e.g., saxitoxin in butter clams) and magnification (e.g.,
             ciguatera) during food-chain transfers can occur and may be important in understanding the fate
             of phycotoxins in the marine environment, although these processes are poorly understood
             (Shimizu, 1987). Shellfish differ markedly in their physiological responses, and in their ability
             to accumulate, metabolize, and eliminate various biotoxins (Shurnway, 1990; Shurnway and
             Cucci, 1987). Therefore, information obtained for one species is not necessarily applicable to
             others.










                                                            17






                                                                          TABLE 2

                        ALGAL SPECIES WHICH POSE A THREAT TO FINFISH, SHELLFISH AND WILDLIFE
                                                                  IN NORTH AMERICA


                             HarmNLAIjgal Species                        G!Mp     hic Area                   Affected Orpnisms
                                                                                                           Mussels, surfclams, softshell
                                                                                                           clams, sea scallops, butterclams,
                                                                                                            ocean quahogs, oysters,
                            Alexan&iwn spp. (PSP)                 Northern Atlantic and Pacific            gasuqx)as, lobsters, crabs
                                                                    Coast of North America                 Herring, salmon, menhaden,
                                                                                                           sandlance, mackerel and possible
                                                                                                               other fish species.
                                                                                                           Whales, sea lions* sea ottere,
                                                                                                                     sea @i;&
                                                                                                           Squid, zgplankton and other
                                                                                                              ben ic in74;;;;VWes
                              Alexand.riwn monilata                      Gulf of Mexico                    Oysters, coquinas, mussels,
                                                                                                                 xastrobods, fish
                           Pseudonitzschia ngens f.              Gulf of Maine; eastern Canada,                      Mussels
                                muldseries tAuS                         Pujqet Sound, WA
                          P. pseudodelicatissitw (ASP)              New Brunswick, Canada                            Mussels
                                P. australis (ASP)                          California                        Anchovies, sea birds
                           Probably P. australis (ASP)                Washington, Oregon                   Razorclame, Dunizeness crabs* .
                               Unidentified (ASP)                         Massachusetts                           Bay scallops*
                                                                              Maine                               Sea scallops*
                             Dinophysis spp. (DSP)                  Nova Scotia, Gulf Of St.                         Mussele
                                                                        Lawrence, Canada
                           Prorocentrum lima (DSP)                    Nova Scotia, Canada                            Mussele
                               Prorocentrum SVD.                       Low Island Sound                    Northern quahogs, bay scallops
                              GyrodEnium aureolum                Northern New England (Maine)               Mussels, softshell clams*
                         Aureococcus anophagefferens                New York, Rhode Island,                   Bay scall    , mussels
                                                                           New Jersey                       Anchoa sp.T,c6docerans
                                                                                                           Bay scallops, surfclams, oysters,
                                                                                                           southern quahogs, coquinas.
                           Gymnotfinium breve (NSP)              Gulf of Mexico, South Atlantic                      Tunicates
                                                                              Bight                           Many commercial and
                                                                                                           recreational species of fish.
                                                                                                           Sea birde, sea turtles, manatees*,
                                                                                                                     dolphins'
                                Chaetoceros spp.                        Pacific northwest                  Salmon aquaculturp, possibly
                                                                                                                  Otner species
                             Heterosigma akashiwo                       Pacific northwest                      Salmon aquaculture
                                                                        Narragansett Bay                            zooplfnkton
                                                                                                           Striped bass, founder, croaker,
                             Unnamed gonyaulacoid                      Mid-Atlantic region                 mullet, menhaden, pififish, sea
                                                                                                                blue crabs, biky scallops
                             GambiercUscus tojdcus                                                         Grouper, snapper, mackerel, jack,
                              Prorocentrum 1hna*                                                           barracuda, parrot fish tan goat
                                  P. concavum*                     South Florida Florida Keys                 fish, &id other hiai;t
                               P. hoffmanmanum-                  Puerto RM, U1 Virgin Islands
                             Ostreopsis lenticuWis*                           vai , Guam                            Gastropods
                                  a. siapwnsis*              I                                       I
                            Found to contain algal toxins, or be adversely attected by marine algae
                            Causative algae implicated, not confirmed.
                                                                                18









             Marine marnmals and wildlife including endangered species are also threatened by toxic algae.
             Over the past few years, PSP toxins transferred through mackerel have been implicated in the
             mass mortality of humpback whales in the Northeast (Geraci et al., 1989); domoic acid
             transferred through anchovies has been implicated in the dealth of brown pelicans and cormorants
             in the Southwest (Work et al., in press; Fritz et al., 1992) and brevetoxins possibly transferred
             through menhaden were implicated in the mass mortality of bottlenose dolphin in the southeast
             (Anderson and White, 1989). The transmission of dinoflagellate toxins through marine food
             chains can also have ecologically significant sub-lethal effects. PSP toxins sequestered by butter
             clams function as an effective chemical defense against important seabird, sea otter and fish
             predators, and may influence the distributions of these species (Kvitek and Beitler, 1991; Kvitek,
             in press). The ecological impact of dinoflagellate toxins in the marine food chain may therefore
             have profound consequences for conservation biology and our attempts to preserve and protect
             endangered species. The scope of this overall problem is unknown.

             Algal blooms may have harmful effects not related to production of toxins, such as oxygen
             depletion of the water column (Ropes et al., 1979), fish suffocation from stimulation of gill
             mucus production, or mechanical interference with filter-feeding structures (Homer et al., 1990).

             The Fisheries and Food Webs working group identified seven major impediments to progress in
             the biology and ecology of toxic shellfish; three impediments in the areas of fish kills and
             aquaculture of finfish; two in the area of the effects of toxins on the marine food web, and five
             in shellfish monitoring programs. The group recommended solutions to these impediments.

                 1.1. Shellfish: Impediments and Recommendations

             IMPEDIMENT: Available information on toxin kinetics (toxin uptake and detoxificationl
             depuration) and anatomical distribution of toxins in shellfish is restricted and limited to a few
             bivalve species.

             RECOMMENDATION: Determine factors controlling accumulation and loss of toxins in
             commercially important inshore and offshore shellfish, including environmental factors,
             characteristics of the phytoplankton assemblage (e.g.,, relative abundance and toxicity of
             implicated algal species), and prior history of exposure to toxins.

             Field studies relating bloom dynamics to shellfish toxicity patterns at the appropriate spatial and
             temporal scales are extremely rare.       This information is necessary to identify potential
             aquacultm species which are less susceptible to accumulation and long-term retention of toxins,
             select suitable indicator species, and evaluate the potential for species-specific closures of
             shellfish harvesting grounds. These data will also allow optimization and streamlining of costly
             monitoring efforts (e.g., determination of optimum sampling frequency) and development of
             mitigation strategies. Field studies correlating phytoplankton and shellfish toxicities in
             combination with experimental toxification studies will allow unequivocal cause-effect linkage
             between shellfish toxicity episodes and their source.



                                                             19










                  Emphasis should be placed on:

                     Understanding reaction products and kinetics of metabolic transformations of toxins in
                     shellfish tissues.


                  Two- to three-year studies in which the history of toxication is well characterized are best suited
                  to meet this objective. Results obtained in the laboratory should be compared with those
                  documented in field populations where the source of toxin may be unknown or poorly
                  characterized. Toxin conversions in shellfish tissues may increase public health risk.          For
                  example, low potency PSP toxins in algal cells are converted to more potent metabolic end
                  products in some bivalve tissues.

                  ï¿½  Anatomical/cytochemical localization and transfer of toxins among tissues, especially
                     when only some tissues are marketed, (e.g., in scallops, surf clams and razor clams).

                  ï¿½  Characterization of magnitude and causes of variability in toxin accumulation within a
                     population (e.g., in relation to body size, reproductive condition, or feeding zone), and
                     among different species.

                  ï¿½  Development of methods to enhance the rate of toxin depuration (detoxification),
                     especially in species of high economic value that are characterized by prolonged toxin
                     retention (e.g., surf clams, butter clams and sea scallops).

                  Treatment of toxic shellfish should involve relatively short time scales compatible with industry
                  needs. Treatment methods include manipulation of natural environmental variables (e.g.,
                  temperature), development of food processing technology, or artificial methods such as treatment
                  with ozone.


                     Determine the relationship between algal population dynamics and seasonal and spatial
                     patterns of toxicity in shellfish populations (e.g., how do vertical distribution of algal
                     cells and benthic re-suspension affect toxin transfer?).

                  This requires high-frequency sampling of shellfish stocks, phytoplankton populations and
                  hydrographic features at selected field sites that are readily accessible and where toxic/noxious
                  blooms are known to be a recurrent problem. Inter-annual variability should be determined.

                     Develop predictive models of toxin kinetics (uptake and depuration by shellfish) based
                     on integration of field and laboratory studies.

                  Simple bioenergetics-based models have been used previously to describe and predict the
                  accumulation of anthropogenic contaminants in aquatic systems. Modeling efforts should be
                  especially useful in identifying the likely source and history of shellfish intoxication in areas
                  where extensive phytoplankton monitoring is unavailable or impractical. Predicting the risk of
                  contamination in areas as yet unaffected by shellfish toxicity episodes, but where the presence


                                                                  20









             of toxic algae has already been documented, may also be possible. Models will likely aid in the
             design of an optimum sampling schedule for the monitoring of a particular shellfish resource.

             IMPEDIMENT: Information is lacking on the relative sensitivities of different life history
             stages to harmful algae, and on the long-term effects of algal toxinslmetabolites on growth,
             reproductive success and recruitment of shel(ftsh populations. Past work has largely focused
             on adult stages of a few species and on short-term effects on individuals.

             RECOMMENDATION: Assess the short- and especially long-term concentration-dependent
             chronic effects of harmful algae on various life history stages of shellfish (e.g., larvae,
             juveniles, adults). Determine the mode of action and effects of toxins on these developmental
             stages at both organismal and population levels.

             Blooms of harmful algae may exert sublethal effects on shellfish populations, and thus affect
             long-term persistence as well as harvestable yields of the resource. For example, toxic
             Alexandrium cells (Shumway and Cucci, 1987; Brice1j et al., 1991) and direct contact with
             Aureococcus anophagefferens cells (Tracey, 1988) can significantly inhibit feeding activity in
             some bivalve species. Field and laboratory studies should assess the relevance of such transient
             physiological effects on population fitness traits (e.g., growth rates and reproductive
             performance), and identify the most critical developmental stages affected. Studies involving
             natural populations will depend on bloom incidence in the field. Mitigation strategies such as
             transplanting of stocks to unaffected areas during a harmful bloom, or modification of culturing
             practices and schedules (e.g., early or delayed planting of seed), and stock rehabilitation efforts
             following a toxic episode could thus be designed to minimize adverse effects. The time frame
             for such studies will depend on the lifespan and growth rate of the species or developmental
             stage under consideration, but useful data could be provided within 2-3 years.

             IMPEDIMENT:           The identity, mode of action, and species-speciftc impacts of
             toxinsImetabolites associated with previously unimplicated harmful algae (e.g., Aureococcus
             anophagefferens, and Gyroilinium aureolum) have not been clearly established.

             RECOMMENDATION: Identify the potentially noxious bioactive compounds associated
             with these algae, determine their mode of action on shellfish, and develop sensitive bioassays
             for their rapid detection.

             Blooms of Aureococcus anophagefferens have recurred in New York waters since 1985, and were
             also documented in RI and NJ waters in 1985 (Cosper et al., 1989). Blooms of a related
             picoplanktonic alga recently occurred in Texas (Stockwell et al., in press). Aureococcus caused
             weight loss of adult bay scallops and mortality of adult mussels and recruitment failure of bay
             scallops (Tracey, 1988; Bricelj and Kuenstner, 1989), but only anecdotal information is available
             for the effects on other commercially important bivalves, such as the American oyster and hard
             clam. Less susceptible species might provide a viable aquaculture alternative during brown tide
             episodes.



                                                             21









                   Gyrodiniwn aureolwn, a species with ichthyotoxic properties, has been shown to cause mortalities
                   in a number of bivalves (scallops, oysters and mussels) in Europe (Tangen, 1977; Partensky and
                   Sournia, 1986) and was recently considered responsible for mass shellfish mortalities in Maquoit
                   Bay, ME (Heinig and Campbell, 1992). Preliminary studies have shown deleterious effects of
                   this alga on feeding of nine juvenile species, and mortality in some species (Shurnway,
                   unpublished data).

                   Medium-term (3 years) laboratory studies are required to determine the effects of these algae on
                   a broad range of species. Such studies should verify the existence of a concentration threshold
                   below which no adverse effects are observed. Understanding of concentration-dependent effects
                   will help to develop mitigation strategies (e.g., site selection for aquaculture ventures). Bioassays
                   could be developed over the short-term (2 years), before chemical characterization of bioactive
                   compounds has been achieved, and should determine if these active compounds are extracellular
                   and/or intracellular.


                   IMPEDIMENT: An extensive historical database on accumulation and depuration of PSP
                   toxins has been collected by state and federal monitoring agencies, but this "grey" literature
                   is scattered and not readily accessible to potential users.

                   RECOMMENDATION: Compile, integrate and interpret existing data in order to further
                   elucidate general patterns of toxification/detoxification in commercially important shellfish
                   on a regional and national basis.

                   IMPEDIMENT: Availability of isolates of toxiclnoxious algae is limited. These are essential
                   for physiological studies on effects and mode of action of toxins.

                   RECOMMENDATION: Establish additional cultures of algal isolates/clones, and develop
                   culture techniques where these are not available (e.g., Dinophysis spp.).

                   For some species of harmful algae that affect the United States, only single cultures are available
                   (e.g., Aureococcus anophagefferens). This severely biases laboratory studies towards isolates that
                   may not be representative of the natural populations. Efforts to culture the DSP-producing
                   Dinophysis Spp. have so far been unsuccessful. Continued research on Dinophysis species is
                   highly desirable, but does not warrant a major investment of funds until DSP is shown to be a
                   real rather than a perceived problem to U.S. shellfish.

                   IMPEDIMENT: Rapid, reliable methods for field-testing of shellfuh are lacking. Standards
                   for quantiflcadon of toxins are limited and often unavailable. Analytical methodsfor detection
                   and quantification of toxins in animal tissue (eg., PSP and DSP toxins) need improvement.
                   Lack of radiolabeled toxic compounds limits the scope of laboratory studies on toxin transfer
                   in shellfish.





                                                                    22









             RECOMMENDATION: Provide low cost, certified toxin standards; develop and test rapid
             methods for in situ detection of toxins; improve analytical methods and provide radiolabeled
             toxins for quantification of toxins in shellfish tissues.

                  1.2. Finfish: Impediments and Recommendations

             IMPEDIMENT: The physiological responses of fish exposed to toxic and other marine
             hannful algae are poorly known.

             RECOMMENDATION: Complete the identification of known or suspected fish-killing algal
             species and their toxins or harmful metabolites. Establish the mechanisms underlying algal-
             caused fish kills, the routes of toxin delivery, and the varying effects on different life history
             stages of fish. Develop standardized laboratory bioassay techniques for key fish species.
             Examine bioaccumulation of toxins and possible physiological feedback mechanisms between
             fish and phytoplankton.

             Although fish exposed to harmful algae are known to die of respiratory failure in some cases,
             the underlying causes and physiological mechanisms may vary among species or are unknown.
              Severe economic damage is caused by several species of harmful     'algae, although the routes of
             toxin delivery and physiological reactions of fish are poorly understood. Modes of toxin
             production and ichthyotoxic action should be studied because human consumers of fish may be
             at risk if toxin accumulates in an unpredictable fashion in fish muscle tissue. This limits the
             ability to mitigate the problem for aquaculture and to understand and predict the consequences
             for wild fish stocks. There may be multiple causes of fish death due to harmful phytoplankton,
             and their relative contribution is difficult to detect or predict. For example, the raphidophyte
             microflagellate Heterosigma akashiwo (found on both coasts of the United States) and related
             species (Chatonella marina in Japan) may suffocate fish due to massive mucus production by the
             gills, by neurotoxin suppression of respiration, or by destruction of blood components (Onoue,
             1990; Chang et al., 1990; Black et al., 1991; MacKenzie, 1991; Rensel Associates and PTI
             Environmental Services, 1991). Tools and methods of investigation are generally available and
             the above recommendations could be achieved in 5 to 7 years.

             IMPEDIMENT: Harmful phytoplankton blooms are a major impediment to the operation and
             development of marine finflish aquaculture.

             RECOMMENDATION: Develop more effective methods of mitigating the effects of
             harmful algal blooms on finfish aquaculture.

             Catastrophic losses of aquaculture fish have occurred in the United States in recent years due to
             species of harmful phytoplankton previously not recognized as toxic or present (Homer et al.,
             1990). Monitoring by fish farmers in coastal waters provides a valuable link and permanent
             sampling platforms for assessing the frequency and trends of harmful blooms. Mitigation based
             on physical movement of water into net-pens or oxygenation is feasible and already practiced in
             some cases, but once the underlying causes of fish mortality are known, other types of mitigation


                                                             23









                  will be possible. Such strategies are necessary for harmful algal species that often occur
                  throughout the water column or are difficult to detect in aerial or boat surveys. The time lines
                  involved in developing effective mitigation techniques are short-term (2-4 years), depending on
                  the species involved.

                  IMPEDIMENT: Investigators of fish-kills involving algal blooms often lack training,
                  specialized equipment, and communication networks needed to detect, investigate, and
                  deteimine causes.


                  RECOMMENDATION: Promote and expand communication among aquatic user groups,
                  resource agencies, and specialists. Develop field sampling protocols and streamlined
                  physiological or pathological assays to determine which algal toxins or species were
                  responsible for fish losses. Establish systematic reporting and data base management of
                  marine fish kill data.

                  Marine user group and resource agency personnel arr. generally not experienced or trained
                  regarding harmful marine phytoplankton blooms. Fish kills often occur quickly, before agency
                  or research personnel are able to react. As a result, little knowledge about the causes of algal-
                  related fish kills has been gained. Although fish kills may be increasing in frequency,
                  determination of actual trends is impossible at present. Many resource agencies will investigate
                  fish kills first for anoxic/hypoxic water conditions, or will suspect diseases, chemical spills or
                  pollution before searching for other causes. Phytoplankton sampling and/or testing for toxin
                  content is often a low priority or is not attempted. Development of handbooks similar to those
                  used to investigate fish kills in freshwater (e.g., Meyer and Barclay, 1990) will allow agency staff
                  or volunteers to collect fish tissues in a proper manner for analyses. Fostering communication
                  between university or federal experts and aquatic user groups (fishermen, aquaculturists) will
                  expedite reporting and accurate assessment of the causes and extent of fish kills. This is a short-
                  term efffort that will require periodic -review.

                      1.3. Food Web Effects: Impediments and Recommendations

                  IMPEDIMENT: Investigative responses to kills of marine animals are often ineffective.

                  RECOMMENDATION: Establish rapid response research capability and associated
                  geographic information system (GIS) data base.

                  Present studies of sudden toxic bloom events and resulting contamination and kills of marine
                  biota are not pre-planned, are poorly coordinated, and often take place too long after the event
                  to be useful. Thus, determination of the causative organism(s), the routes of toxin transfer, the
                  nature of the effects on marine animals, and the risks to public health is difficult or impossible.
                  As such, these responses are usually reactive, not proactive. By the time stricken animals arrive
                  at analytical facilities, toxins may no longer be present at detectable levels. Infrastructure and
                  funding should be provided to enable the investigation of toxic and noxious blooms at their peaks
                  in terms of toxin assays of plankton, intermediate vectors, fish, and other wildlife. Selected


                                                                   24









             species should be assayed over time to determine duration of toxin retention and changes in toxin
             profiles. Professionals'in the fields of wildlife veterinary medicine and biology, and toxin
             analysis, should be trained in sampling protocols aimed specifically at marine biotoxins.
             Regional rapid response teams should be coordinated with the simultaneous efforts of similar
             teams investigating blooms and the origin and fate of the toxins (see below). All results could
             be stored as overlays in a GIS data base. This type of archive and retrieval system offers the
             most effective means of linking spatially-related data and testing hypotheses about the importance
             of oceanographic processes, land-use practices, and environmental factors to harmful algal bloom
             dynamics and food web effects.

             IMPEDIMENT: Risks of biotoxins to marine animals or to public health resulting from the
             movement of toxins through the marine food web chain cuffently cannot be addresseil.

             RECOMMENDATION: Develop quantitative models of the fate and consequences of
             biotoxins in the marine food web.


             Important pathways of toxin transmission have not been identified, nor has toxin accumulation
             potential and sensitivity been determined for many key marine species (i.e., species that are either
             commercially important, endangered, or serve as major food web links). The information
             necessary for development of these models could be obtained primarily by multi-disciplinary,
             rapid response groups investigating bloom dynamics and toxin transmission in the food web,
             coordinated with other supporting studies (e.g., laboratory and field feeding and observational
             studies).

                2. Shellfish Monitoring ProgranLs

                     2.1. Background

             In the United States, the monitoring of marine biotoxins and/or associated phytoplankton in
             seafood and the environment has been primarily the responsibility of state-sponsored programs.
             The relative success of these programs can be measured by the lack of overt public health
             morbidity or mortality from consuming contaminated seafood (Ahmed, 1991; Bean et al., 1990).
             However, the sampling programs of individual states vary in magnitude (Table 3), dependent
             upon the degree of seafood production or import, commitment of resources, and political will.

             The responsibility of the state programs can extend to products from international waters and
             interstate imports. These programs can implement the closure of a fishery based upon existing
             action limits for only two marine biotoxins, paralytic shellfish toxin and amnesic shellfish toxin,
             or they can exercise other policy considerations such as the lack of sufficient information about
             a fishery. Presently, closure limits based on the presence of other marine biotoxins have not been
             established. When few monitoring stations are employed to cover large coastal areas and the
             frequency of sampling is relatively low, program officials close large fishery areas as a
             conservative measure to protect the consumer and consumer confidence in the seafood industry.
             This is the case especially when health officials attempt to deal with toxic effects from an


                                                              25











                                                              TABLE 3


                                          1989 STATE PSP MONITORING STATIONS


                                             STATE             NUMBER OF SAMPLES
                                                                    (NO. STATIONS)
                                              Maine                       3500 (200)
                                           Washington                      1900   (50)
                                            California                     1100   (not fixed)
                                             Oregon                          780  (16)
                                          Massachusetts                      700  (98)
                                              Alaska                         654  (40)
                                           Connecticut                        51   (5)_
                                          Rhode Island                        40    (8)
                                        New 11arnpshire                      34 (1)


                  unidentified toxic substance. Monitoring    is not only important in the closure process, but is
                  critical for establishing re-openings.

                  International imports are subject to routine inspection by United States Customs and can be
                  embargoed if found to be contaminated. On the other hand, international exportation of seafood
                  products from the United States are subject to the seafood importation standards at the point of
                  destination.


                  The collection of a sample can be accomplished in many ways. Sub-samples can either be taken
                  by program personnel, delivered from commercial harvests at the point of landing, or before
                  delivery to market. State programs support "sentinel" species programs whereby specific or
                  mixed species of molluscs (e.g., mussels for PSP) are strategically placed along coastal areas that
                  are subject to harvesting activity. These sites are sampled regularly to provide an early warning
                  of the presence of toxins. The relative effectiveness of a sentinel program can be measured by
                  the number of sampling sites per area covered and the frequency of sampling. In addition, with
                  few exceptions, sentinel sites typically are positioned near-shore and do not provide information
                  about offshore toxin distribution. In practically all cases, the analyses for marine biotoxins are
                  carried out by state or federal laboratories, the exception being the availability of uncertified
                  commercial laboratories that conduct analyses for domoic acid.
                                      I                                                         I




























                  Within state programs, however, the responsibility of seafood monitoring can be decentralized,
                  with different programs charged with the management of different seafood industries (e.g.,


                                                                  26









             shellfish industry, finfish industry, crab industry, etc.). The specific mandates of each of these
             sub-programs may present a narrow focus to a particular biotoxin problem, depending upon the
             region and indigenous biotoxins present. While focused programs may provide adequate
             protection to the consumer, these efforts can be inflexible when attempting to respond to the
             presence of newly-recognized biotoxin threats. Communications between state programs and
             interagency entities can at times be tenuous, even during crises.

             Ideally, these efforts are designed to lead to a proactive response. 11at is, the toxins are detected
             within the seafood prior to commercial sale of product. In practice, this may be viewed as
             reactive, since a toxic sentinel site or product is already contaminated and the monitoring effort
             is not 100% for all seafood (nor should it be). The State of Florida exercises a "preemptive"
             monitoring program, which combines seafood and phytoplankton monitoring with aspects of
             remote sensing to recognize algal blooms prior to their impact on fisheries.

                       2.2. Impediments and Recommendations

             IMPEDIMENT: Current monitoring efforts are too limited and inflexible to measure the full
             impact of marine biotoxins. The extent of the distribution of marine biotoxins in consumed
             seafood products is not known.

             RECOMMENDATION: In those instances where newly-recognized toxins impact a flshery,
             state and federal resources should be made available to aid monitoring programs. Various
             seafood products should be surveyed on a region-by-region basis for the most important
             biotoxins and the products affected.

             Existing routine monitoring programs are, by design, limited to managing specific resources. On
             those occasions where marine biotoxin crises arise, routine state and federal funding mechanisms
             are inadequate to address the immediate concerns. This places a huge burden on state and federal
             agencies to redirect resources from predetermined programs to deal with the crisis. Contingency
             funds and a mechanism for rapid deployment are needed to help state and federal programs
             respond to crises such as toxic plankton blooms, fish kills, and marine mammal kills. Time is
             of the essence if these efforts are to be effective.


             All coastal areas contain various types of marine life and biotoxins. The utilization of existing
             biotoxin monitoring elements to address all biotoxin concerns may not be appropriate. Tberefore,
             the search for affected seafood products and additional biotoxins will be an evolving process.
             The ultimate goal of this effort will be to create more efficient and cost-effective programs.

             IMPEDIMENT: The public health community and seafood industry require early warnihg of
             toxiclhar7nful phytoplankton blooms to protect seafood consumers and producers.

             RECOMMENDATION: Identify the best indicator species within specific regions. Identify
             the best sites and sampling strategies for these indicator species.



                                                              27









                  Presently, bivalve molluscs (e.g., mussels, oysters or clams) are employed as the primary sentinel
                  organisms. These species may or may not reflect accurately the true nature of a toxic algal
                  bloom. For example, mussels have been employed as sentinel organisms for the detection of
                  domoic acid (Haya et al., 1991). They may not, however, always be the best predictor of the
                  presence of domoic acid, as was shown on the West Coast when razor clams were toxic but
                  mussels were not (Wood and Shapiro, 1992). A primary sentinel species may, in fact, be a non-
                  consumed species with characteristics of toxin uptake, distribution, and retention that provide
                  useful data for particular toxins. Further studies on a regional basis are required to determine
                  which species best accommodate specific biotoxins and analyses. These studies should be
                  conducted locally to account for regional variations and should identify species with both long-
                  and short-term retention times. These studies would take 2-3 years for completion.

                  Monitoring programs are only as good as the analytical support they receive. Present laboratory
                  support is limited by the number of facilities and methods currently available. In addition, in
                  those instances where newly-recognized toxins are found, a full understanding and
                  characterization of the toxin must be obtained before an appropriate sentinel organism can be
                  identified.


                  In order to identify sites and sampling strategies, models that describe temporal and spatial
                  distributions of toxins must be developed. The presence of biotoxins in marine life is subject to
                  the effects of regional hydrographic conditions. In order to develop the model and account for
                  the environmental conditions, the distribution of toxins among sites and individual organisms
                  must be evaluated statistically and include both nearshore and offshore locations.

                  IMPEDIMENT: The collection, preservation, and handling of naturally-occurring toxic
                  seafood and algal samples for use by researchers and public health officials is hampered by
                  a lack of standarifted procedures.

                  RECOMMENDATION: Develop internationally accepted and appropriate collection,
                  preservation, and shipping protocols.

                  With the lack of standardized handling and shipping methods, the implementation of public health
                  measures is, hampered. Furthermore, the development and refinement of methods for biotoxin
                  detection are also impeded. In particular, it is necessary to understand the fate and stability of
                  biotoxins in natural seafood matrices. Currently, the methods used for the collection and handling
                  of contaminated seafood are not standardized. It is not known how these conditions affect toxin
                  stability. If sample integrity is compromised, analytical results are questionable. This could be
                  easily tested by conducting spiking studies or splitting contaminated samples prior to shipment.
                  The initial method development and subsequent collaborative studies could be completed within
                  2-3 years.

                  Due to increasing interest and awareness of marine toxins and algal blooms on the part of the
                  general public, scientists, and public health officials, we can assume that in the future, new toxins
                  or the extension of known toxins to new seafood items will be documented. As the public and


                                                                   28









             health professionals inevitably begin to associate illness with the consumption of a seafood
             product, there will be increasing demands to "explain the cause of the sickness". There is a need
             for the development of standardized, generic laboratory procedures, techniques, protocols, and
             surveys that could be compiled and be "ready-to-use" by local and state public health agencies
             to meet and deal with these inquiries.

             IMPEDIMENT: Slightly different policies for dealing with fishery closures due to a marine
             toxin outbreak can lead to industry problems in contiguous states sharing the coastline and
             the toxin problem.

             RECOMMENDATION: Encourage the states to form regional communication networks
             and harmonize fisk management policies.

             If one state closes a fishery while the others do not, processors and fishermen move from the
             affected state to the state where the fishery is still open. This places stress on the open industry
             and appears to place the closed state at a competitive disadvantage. In addition, if the toxin
             outbreak does impact several states, regulatory tracking of risk species becomes virtually
             impossible due to lengths of coastline, mobility of fishermen, and multi-state licensing of fishing
             boats. It is better if states sharing coastline and toxin risks develop a harmonized closure action
             plan.

             IMPEDIMENT: Authority for dealing with both known and new marine toxins occurring in
             seafood is sometime fragmented within state management agencies, leading to inter- and intra-
             agency jurisdictional overlap and confusion.

             RECOMMENDATION: Encourage the states to streamline programs dealing with marine
             toxin risks by placing monitoring and management control in a single regulatory agency.

             In many states there is a fragmentation of authority between state agencies for action on a fishery
             due to the presence of a toxin. For example, in some states, authority is divided among the
             Departments of Agriculture (for crustacea and finfish), Health (molluscan shellfish), and Fisheries
             (resource closures). There may be a variety of good reasons for this fragmentation; nonetheless,
             when a toxin such as PSP is found in a "non-traditional" species such as Dungeness crab, it
             becomes difficult for the state to formulate and take appropriate action. States should be
             encouraged to streamline their programs in dealing with marine toxin risks, i.e., place monitoring
             and control in one agency.











                                                             29












                   V. SUMMARY


                   The preceding text lists numerous impediments to progress in the area of marine biotoxins and
                   harmful algae and addresses each with a series of recommendations. The length of these lists
                   defines the major challenge before us if the goal of the National Plan is to be realized. Some
                   might conclude from the many impediments that past research has made little progress, but this
                   is certainly not the case. The scientific and policy disciplines involved are healthy and maturing,
                   though underfunded relative to the expanding problem.

                   The rate and extent of progress from here will depend in large part on how effectively the
                   recommendations in this National Plan are implemented. Our hope is that numerous state and
                   federal agencies will use this document to identify topics that relate to their particular
                   responsibilities or purviews, and that scientists and private industry will use these ideas to guide
                   their activities as well. No single agency can address all of the identified impediments, but most
                   can be covered by the combined efforts of several organizations. Overlap and omissions are
                   likely however, unless further coordination is attempted at the agency level. The network has
                   been established to make this possible, but concerted efforts will be necessary to keep the lines
                   of communication and coordination open.



























                                                                   30










             VI. ACKNOWLEDGEMENTS

             The workshop and the preparation of this report were supported by the National Marine Fisheries
             Service through a Saltonstall-Kennedy grant (No. NA27FDO092-01 to DMA), by the Southeast
             Fisheries Science Center's Charleston Laboratory, and by the NOAA Coastal Oceans Program,
             which provided travel support for some participants. We gratefully acknowledge the efforts of
             the NMFS Southeast Fisheries Science Center's Marine Biotoxins Group in Charleston SC whose
             local logistical support made the workshop a success. We also acknowledge the contributions
             of colleagues who did not attend the workshop, but who nevertheless provided comments on the
             draft report. Special thanks go to Dean, Jacobson for his contribution of artwork for the cover.





































                                                           31











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                                                               34









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                                                           37










                 Wright, J. L. C., R. K. Boyd, A. S. W. De Freitas, M. Falk, R. A. Foxall, W. D. Jamieson, M.
                 V. Laycock, A. W. McCulloch, A. G. McInness, P. Odense, V. P. Pathak, M. A. Quilliam, M.
                 A. Ragan, P. G. Sim, P. Thibault, J. A. Walter, M..Gilgan, D. J. A. Richard and D. Dewar.
                 1989. Identification of domoic acid, a neuroexcitatory amino acid in toxic mussels from eastern
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                 262, Washington, DC, pp. 207-214.






































                                                               38












             VIII. WORKSHOP AGENDA



             Tuesday, Avril 21, 1992



             8:30         Registration; coffee

             9:30         Announcements - Dr. Sylvia Galloway

             9:15         Welcome - Dr. Robert Kifer


             9:30         Workshop Overview - Dr. Donald Anderson

             9:45         International Perspective - Dr. Donald Anderson
             9:55         NMFS Perspective - Dr. Sylvia Galloway
             10:05        Coastal Oceans Program Perspective - Dr. Leon Cammen
             10:15        FDA Perspective - Dr. Sherwood Hall
             10:25        NIEHS Perspective - Dr. Daniel Baden

             10:35        Break


             10:50        Define National Program; Develop objective statement for
                          the workshop

             12:00        Lunch


             1:15         Assignment to Working Groups; Charge by Co-Chair
             1:30         Working Group Discussion; Goal to generate consensus document in assigned
                          topic

             5:00         Social gathering at picnic area, Marshlands House

             6:00         Dinner - TROGMORE STEW"
















                                                           39













                  Wednesday. April 22, 1"2

                  8:30        Coffee; Working Group discussion continued

                  12:00       Lunch

                  2:00        Plenary Presentations; Pharmacology/Epidemiology/Toxicology - Dr. Daniel
                              Baden
                  2:30        Toxin Analysis/Assays/Chemistry Standards - Dr. Jack Wekell
                  3:00        Shellfish Monitoring - Dr. Richard Danielson
                  3:30        Plankton Monitoring - Dr. Rita Homer

                  4:00        Break


                  4:15        Foodweb Effects - Dr. Alan White
                  4:45        Shellfish Depuration/Physiology - Dr. Sandra Shurnway

                  6:00        Dinner on your own



                  Thursday. April 23, 1992

                  8:30        Coffee; Plenary Presentations; Fish Mortalities - Mr. Jack Rensel

                  9:00        Bloom Biology/Ecology - Dr. Donald Anderson
                  9:30        Remote Sensing - Dr. Patricia Tester
                  10:00       Nutrient/Pollution Effects - Dr. Theodore Smayda

                  10:30       Break


                  10:45       Taxonomy/Genetics/Population Biology - Dr. Karen Steidinger
                  11:15       Hydrography/Physical Oceanography - Dr. Donald Anderson
                  11:45       Discussion: Report structure

                  12:00       Lunch


                  1:30        Working Groups meet to finalize reports

                  4:00        Discussion: Management (structure), networking, funding sources, collaborations

                  6:00        Dinner on your own; Evening Working Group writing (if necessary)



                                                              40













            Friday. April 24, 1992

              8:30       Coffee; Reports by three Working Groups - Strawman priority lists, Executive
                         Summary

              9:00       National Program Priorities Set, Discussion -
                         Dr. Donald Anderson/Dr. Sylvia Galloway

            10:30        Break


            12:00        Workshop Adjourned

            1:30         Informal Lab Tour - John Babinchak/Fran Van Dolah





































                                                         41











                   IX. PARTICIPANTS



                   Dr. Donald Anderson (Co-chair)                        Dr. Sherwood Hall
                   Biology Department                                    U.S. Food and Drug Administration
                   woods Hole Oceanographic Institute                    200 C Street, SW (IHFF-521)
                   Woods Hole, MA 02543                                  Washington, DC 20204
                   (508) 457-2000, Ext. 2351                             (202) 205-4818 OR (202) 254-3888
                   FAX: (508) 457-2169                                   FAX: (202) 254-3982

                   Dr. Daniel G. Baden                                   Dr. Rita Homer
                   marine and Freshwater Biomedical Sciences             School of Oceanography, WB-10
                   Center                                                University of Washington
                   University of Miami                                   Seattle, WA 98195
                   4600 Rickenbacher Causeway                            (206) 543-8599; FAX: (206) 543-0275
                   Miami, FL 33149-1098
                   (305) 361-4738; FAX: (305) 361-4711                   Mr. John W. Hurst
                                                                         Fisheries and Health Science Division
                   Dr. Monica Brice1j                                    Department of Marine Resources
                   Marine Sciences Research Center                       West Boothbay Harbor, ME 04575
                   SUNY at Stony Brook                                   (207) 633-5572; FAX: (207) 633-7109
                   Stony Brook, NY 11794-5000
                   (516) 632-8663; FAX: (516) 632-8820                   Mr. Raffael Jovine
                                                                         Department of Biological Sciences
                   Dr. JoAnn Burkholder                                  University of California at Santa Barbara
                   N.C. State University                                 Santa Barbara, CA 93106
                   Department of Botany, Box 7612                        (805) 893-4319; FAX: (805) 893-4724
                   Raleigh, NC 27695
                   (919) 515-2726; FAX: (919) 515-3436                   Dr. Rikk G. Kvitek
                                                                         Moss Landing Marine Laboratories
                   Dr. Richard Danielson                                 P.O. Box 450
                   Department of Health Services                         Moss Landing, CA 95039
                   Sanitation and Radiation Laboratory                   (408) 633-5606; FAX: (408) 633-5642
                   2151 Berkeley Way, Room 465
                   Berkeley, CA 94704                                    Dr. Roy E. Martin
                   (510) 540-2201; FAX: (510) 540-2053                   National Fisheries Institute
                                                                         1525 Wilson Boulevard, Suite 500
                   Dr. Sylvia B. Galloway (Co-chair)                     Arlington, VA 22209
                   Southeast Fisheries Science Center                    (703) 524-8883; FAX: (703) 524-4619
                   National Marine Fisheries Service, NOAA
                   P.O. Box 12607, 217 Fort Johnson Road                 Dr. John S. Ramsdell
                   Charleston, SC 29422                                  Division of Marine Biomedical and
                   (803) 762-1200, FAX: (803) 762-1998                   Environmental Sciences
                                                                         Medical University of South Carolina
                                                                         Charleston, SC 29425
                                                                         (803) 793-7988; FAX: (803) 792-0664



                                                                    42












              Mr. Jack Rensel                                          Dr. Jack Wekell
              School of Fisheries, HF-15                               Northwest and Alaska Fisheries Science Center
              University of Washington                                 National Marine Fisheries Service, NOAA
              Seattle, WA 98195                                        Seattle Laboratory
              (206) 524-6331; FAX: (206) 524-6331                      2725 Montlake Blvd., E.
                                                                       Seattle, WA 98112
              Dr. Yuzuru Shimizu (not present, but                     (206) 442-7746; FAX: (206) 553-4304
              contributed)
              Department of Pharmacognosy                              Dr. Marleen M. Wekell
              College of Pharmacy                                      U.S. Food and Drug Administration
              University of Rhode Island                               P.O. Box 3012
              Kingston, RI 02881-0809                                  Bothell, WA 98041-3012
              (401) 792-2751; FAX: (401) 792-2181                      (206) 483-4902; FAX: (206) 483-4996

              Dr. Sandra E. Shurnway                                   Dr. Alan W. White
              Bigelow Laboratory for Ocean Sciences                    Northeast Fisheries Science Center
              West Boothbay Harbor, ME 04575                           National Marine Fisheries Service, NOAA
              (207) 633-2173; FAX: (207) 633-6584                      Woods Hole, MA 02543
                                                                       (508) 548-5123; FAX: (508) 548-5124
              Dr. Theodore J. Smayda
              Graduate School of Oceanography                          Dr. Jeffrey L.C. Wright
              University of Rhode Island                               National Research Council of Canada
              Kingston, RI 02881                                       Institute for Marine Biosciences
              (401) 792-6171; FAX: (401) 792-6682                      1411 Oxford Street
                                                                       Halifax, Nova Scotia B3H 3ZI
              Dr. Karen Steidinger                                     CANADA
              Florida Marine Research Institute                        (902) 426-8275; FAX: (902) 426-9413
              Florida Department of Natural Resources
              100 Eighth Ave., SE
              St. Petersburg, FL 33701                                 Revresentine the NMFS Southeast
              (813) 896-8626; FAX: (813) 823-0166                      F"isheries Science Center:


              Dr. Patricia A. Tester                                   Mr. John A. Babinchak
              Southeast Fisheries Science Center                       Dr. Gregory J. Doucette
              National Marine Fisheries Service, NOAA                  Ms. Jeanne D. Joseph
              Beaufort Laboratory                                      Dr. Robert Kifer
              Beaufort, NC 28516                                       Dr. Peter Moeller
              (919) 728-8792; FAX: (919) 728-8784                      Dr. Frances M. Van Dolah


              Dr. Donald Tindall                                       Southeast Fisheries Science Center
              College of Science                                       National Marine Fisheries Service, NOAA
              Department of Botany and Plant Biology                   P.O. Box 12607, 217 Fort Johnson Road
              Southern Illinois University                             Charleston, SC 29422
              Carbondale, IL 62901                                     (803) 762-1200; FAX: (803) 762-1998
              (618) 536-2331; FAX: (618) 453-3441





                                                                 43












                 Representine NOAA Coastal Oceans Program:

                 Dr. Leon Cammen
                 National Sea Grant College Program, NOAA
                 R/ORI
                 1335 East-West Highway
                 Silver Spring, MD 20910
                 (301) 713-2435; FAX: (301) 713-0799












































                                                             44







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                           Biotoxin Workshop Participants (left to right): Sylvia Galloway, Rafael Jovine, Peter Moeller, Fran
                           VanDolah, Jack Rensell, Alan White, Monica Bricelj, Sherwood Hall, John Hurst, Pat Tester, John
                           Ramsdell, Rita Horner, Jack Wekell, Rikk Kvitek, Marleen Wekell, Rick Danielson, Jeff Wright, Ted
                           Smayda, Greg Doucette, Dan Baden, Sandy Shumway, Jeanne Joseph, Don Tindall, Bernie Lanoue,
                           Karen Steidinger, JoAnn Burkholder, Don Anderson (Leon Cammen and Roy Martin, not shown -
                                                                                                                    UN-1
















                           early departure)






                                                                                                                                           - ----------







                                                 DOCUMENT LIBRARY

                                                       March 11, 1991

                                     Distribution Listfor Technical Report Exchange
               Attn: Stella Sanchez-Wade                               Pell Marine Science Library
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               Library, Mail Code C-075C                               Narragansett, RI 02882
               La Jolla, CA 92093                                      Working Collection
               Hancock Library of Biology &                            Texas A&M University
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               Los Angeles, CA 90089-0371                              Gloucester Point, VA 23062
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                                                                                                          Mac 90-32











                  50272-101
                  REPORT DOCUMENTATION 1 1. REPORT NO.                                                              2.                             3. Recipient's Accession No.
                                PAGE                                    WHOI-93-02
                  4. Title and Subtitle                                                                                                            5. Report Date
                  MarineBiotoxins and Harmful Algae: A National Plan                                                                               6. January 1993

                  7. Author(s)                                                                                                                     8. Performing Organization Rept. No.
                  Donald M. Anderson, Sylvia B. Galloway and Jeanne D. Joseph                                                                        WHOI 93-02
                  9. Performing Organization Name and Address                                                                                      10. ProjecVTasWWork Unit No.

                  The Woods Hole Oceanographic Institution                                                                                         11. Contract(C) or Grant(G) No.
                  Woods Hole, Massachusetts 02543                                                                                                  (C) NA27FDO092-01

                                                                                                                                                   (G)

                  12. Sponsoring Organization Name and Address                                                                                     13. Type of Report & Period Covered
                  National Marine Fisheries Service Saltonstall-Kennedy Grant No. NA27FDO092-01,                                                      Technical Report
                  National Marine Fisheries Service's Charleston Laboratory and by the NOAA Coastal                                                14.
                  Oceans Program.
                  15. Supplementary Notes
                  This report should be cited as: Woods Hole Oceanog. Inst. Tech. Rept., WHOI-93-02.



                  16. Abstract (Limit: 200 words)



                  Marine biotoxins and harmful algae represent a significant and expanding threat to human health and fisheries resources
                  throughout the U.S. This problem takes many forms, ranging from massive "red tides" or blooms of cells that discolor the
                  water to dilute, inconspicuous concentrations of cells noticed only because of the harm caused by the highly potent toxins
                  those cells contain. Impacts include mass mortalities of wild and farmed fish, human intoxications and death from contami-
                  nated shellfish or fish, alterations of marine trophic structure, and death of marine mammals, seabirds, and other animals. The
                  nature of the problem has changed considerably over the last two decades in the U.S. Where formerly a few regions were
                  affected, now virtually every coastal state is threatened, in many cases over large geographic areas and by more than one
                  harmful species. The U.S. research, monitoring, and regulatory infrastructure is not adequately prepared to meet this expand-
                  ing threat. In an effort to surmount these problems, a workshop was convened to formulate a National Plan for the prediction,
                  control, and mitigation of the effects of harmful algal blooms on the U.S. marine biota. This report summarizes the status of
                  U.S. research knowledge and capabilities, and identifies areas where research funds should be directed for maximum benefit.







                  17. Document Analysis     a. Descriptors


                  1. marine biotoxins
                  2. harmful algae blooms
                  3. red tides


                  b. ldentifierstOpen-Ended Terms







                  c. COSATI Field/Group
                  18. Availability Statement                                                                        19. Security Class (This Report)                  21. No. of Pages
                  Approved for publication; distribution unlimited.                                                        UNCLASSIFIED                                    59
                                                                                                                    20. Security Class (This Page)                    22. Price


             (See ANSI-Z39.18)                                                          See Instructions on Reverse                                                   OPTIONAL FORM 272 (4-77)
                                                                                                                                                                      (Formerly NTIS-35)
                                                                                                                                                                      Department of Commerce





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