S - Woods Hole Oceanographic Institution

Transcription

AGENDA, ABSTRACTS, & PARTICIPANTS
OCTOBER 27 – 31, 2013
S
Seventh SympoSium on harmful algae in the u.S.
SympoSium ChairS
Leanne Flewelling, Florida Fish and Wildlife Conservation Commission
Barbara Kirkpatrick, Mote Marine Laboratory
loCal organizing Committee
Alina Corcoran, Florida Fish and Wildlife Conservation Commission
Kate Kohler, Mote Marine Laboratory
Vincent Lovko, Mote Marine Laboratory
Bill Richardson, Florida Fish and Wildlife Conservation Commission
Karen Steidinger, Florida Fish and Wildlife Conservation Commission
Steering Committee
Lorraine Backer, CDC National Center for Environmental Health
Greg Doucette, NOAA National Ocean Service
Deana Erdner, University of Texas at Austin
Sonia Joseph Joshi, NOAA Center of Excellence for Great Lakes and Human Health
Judy Kleindinst, Woods Hole Oceanographic Institution
Raphael Kudela, University of California, Santa Cruz
Kathi Lefebvre, NOAA Northwest Fisheries Science Center
Dennis McGillicuddy, Woods Hole Oceanographic Institution
Hans Paerl, University of North Carolina at Chapel Hill
Tammi Richardson, University of South Carolina
Marc Suddleson, NOAA National Ocean Service
SeSSion ChairS
ORGANIZERS
HABS in a Changing World: Raphael Kudela
Bloom Dynamics and Ecology: Quay Dortch and Greg Doucette
Bloom Prediction, Forecasting, and Modeling: Dennis McGillicuddy
Monitoring and Management: Rob Magnien and Vincent Lovko
HAB Genomics and Other ‘Omics: Diane Greenfield
Toxicity and Pathogenicity: Kathi Lefebvre
Fisheries and Foodwebs: Richard Pierce
Human and Animal Health: Jan Landsberg
Prevention, Control, and Mitigation: Marc Suddleson
Human Dimensions of HABs: Sonia Joseph Joshi
Speed Talk Sessions: Leanne Flewelling, Barbara Kirkpatrick, and Alina Corcoran
Logo Design: Eli Minaya, Ringling College of Art and Design
Program Design: Mary Ellen Seyle, designWorks advertising associates
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October 2013
SympoSium SponSorS:
Beagle Bioproducts, Inc.
Chiles Restaurant Group
Florida Fish and Wildlife Conservation Commission
Fluid Imaging Technologies, Inc.
Jelks Family Foundation
McLane Research Laboratories, Inc.
Mote Marine Laboratory
Mote Scientific Foundation
Saigene Biotech, Inc.
START
US Food and Drug Administration
Visit Sarasota County
Student Support:
SPONSORS
Fluid Imaging Technologies, Inc.
National HAB Office
The Phycological Society of America
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Seventh Symposium on Harmful Algae in the U.S.
Seventh SympoSium on harmful algae in the u.S.
October 27 – 31, 2013
Sarasota, Florida
Agenda, Abstracts, & Participants
Table of Contents
Organizers _________________________________________________ 1
Sponsors __________________________________________________ 2
Agenda Overview ___________________________________________ 4
Symposium Agenda _________________________________________ 7
Poster Schedule ___________________________________________ 17
Keynote Address ___________________________________________ 27
Abstracts of Oral Presentations _______________________________ 29
ORAL
TABLE
PRESENTATIONS
OF CONTENTS
Abstracts of Speed Talk Presentations __________________________ 85
Abstracts of Poster Presentations_____________________________ 123
Author Index _____________________________________________ 197
Symposium Participants ____________________________________ 207
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October 2013
meal information
Breakfast: 7:00 am – 8:30 am
Breakfast will be served each day (Monday through Thursday) in the Boathouse
(behind the hotel).
lunch: 12:30 pm – 2:30 pm
Lunch coupons provided to all registered attendees (one per day, Monday
through Thursday) can be used to purchase food (up to $10 per coupon) in the
hotel restaurant or at sandwich carts located outside. Seating will be available in
the Boathouse and on the Conference Center patio.
dinner is on your own (Monday through Wednesday).
In addition to Sunday afternoon registration hours, the registration desk will be staffed
each day from approximately 7:30 am to 2:00 pm for information, T-shirt and poster
sales, and other needs.
All talks and poster sessions will be in the Sarasota Ballroom.
Sunday, oCtoBer 27
12:00 pm - 5:00 pm
1:00 pm - 5:00 pm
4:00 pm - 6:00 pm
6:00 pm - 9:00 pm
Registration - Convention Center Registration Desk
Young Investigators’ Symposium - Florida Room
NHC Meeting - Boardroom
Welcome Reception - Florida Room and Poolside
AGENDA
ORAL PRESENTATIONS
OVERVIEW
evening aCtivitieS
Tuesday October 29 8:00 pm – 9:30 pm; Sarasota Ballroom
Roundtable: Methods for Evaluating HAB Species and Toxins: Progress, Comparative
Studies, and Meeting End-User Needs
Organizers:
Dianne I. Greenfield (Univ. of South Carolina) and Kathryn J. Coyne
(Univ. of Delaware)
The goal of this forum is to discuss the advancements and challenges of developing
various methods and technologies for evaluating HAB assemblages and toxins, including
cross-comparison studies.
The objectives of this roundtable discussion are to 1) identify the most important
criteria end-users consider when selecting a particular HAB monitoring or research
approach (e.g., cost, accuracy, sample through-put, etc.), 2) highlight progress on a range
of methods for HAB species and toxin detection and quantification, 3) discuss outcomes
and challenges faced while performing cross-comparative evaluations, and 4) evaluate
the capacity for integrating different technologies within HAB monitoring and research
networks.
Thursday, October 31 6:00 pm – 8:30 pm; Sarasota Ballroom
Symposium Banquet and Student Award Announcements
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Mon
October 28
Tue
October 29
Wed
October 30
Thu
October 31
Breakfast 7:30-8:30
Opening Remarks
8:30-9:45
Plenary
Oral Session 4
Prevention, Control,
and Mitigation
Oral Session 7 HAB
Genomics and Other
'Omics
Oral Session 10
Bloom Prediction,
Forecasting, and
Modeling
Break 9:45-10:15
Oral Session 1
HABs in a Changing
World
10:15-12:30
Poster Session 2
Oral Session 8
Human and Animal
Health
Break 11:15-11:30
Oral Session 2
Human Dimensions
of HABs
Speed Talks 3
Break 11:15-11:30
Oral Session 5
Monitoring and
Management (1)
Poster Session 4
Poster Session 5
Lunch 12:30-2:00
2:00-3:15
Oral Session 3
Bloom Dynamics and
Ecology (1)
Oral Session 6 HAB
Toxicity and
Pathogenicity
Break 3:15-3:45
Speed Talks 1
Speed Talks 2
Town Hall
Meeting
(2:45-4:45)
Oral Session 11
Bloom Dynamics and
Ecology (2)
Break 3:15-3:45
AGENDA OVERVIEW
3:45-4:45
Oral Session 9
Monitoring and
Management (2)
Oral Session 12
Fisheries and
Foodwebs
Closing Remarks
Evening
Activities
Poster Session 1
and social mixer
5:00-6:30 pm
Poster Session 3
and social mixer
5:00-6:30 pm
6:00-8:30pm
Banquet
(Ballroom)
8:00-9:30 pm
Methods Round
Table (Ballroom)
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October 2013
SYMPOSIUM
AGENDA
ORAL
PRESENTATIONS
SYMPOSIUM
AGENDA
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October 2013
MONDAY, OCTOBER 28, 2013
8:30
Opening Remarks
9:00
Plenary, Karen A. Steidinger: Dinoflagellate HAB science: Then and now
9:45-10:15
Break
oralS SeSSion 1: haBS in a Changing World
time
presenter
Chair, raphael Kudela
title
10:15 Christopher
Gobler
The role of coastal ocean acidification in promoting and exacerbating the
effects of harmful algal blooms
10:30 Hans Paerl
Managing harmful cyanobacterial blooms along the freshwater-marine
continuum in a world experiencing human and climatically-mediated
change
10:45 Patricia Tester Gambierdiscus biogeography and the consequences of climate change
11:00 Ann Jochens
Harmful algal bloom integrated observing system for the Gulf of
Mexico
11:15-11:30
Break
oralS SeSSion 2: human dimenSionS of haBS
Chair, Sonia Joseph Joshi
Freshwater management policy update: Implementing the 3rd pillar of
the CWA, waterbody management, to form a systems approach
11:45 Porter
Hoagland
CHANS: Florida red tides and coastal populations as a coupled
nature-human system
12:00 Jamie Studts
Perceptions of beach activities during Florida red tide blooms:
A conjoint analysis
12:15 Dianne Shipley Coordinating Florida red tide communication: A case study
12:30-2:00
Lunch oralS SeSSion 3: Bloom dynamiCS and eCology - part 1
Chair, Quay dortch
2:00
Cynthia Heil
Nutrients supporting Karenia brevis blooms: Results of the ECOHAB:
Karenia program
2:15
Pat Glibert
A new suite of nutrient kinetic relationships for picoplankton blooms:
Non-traditional relationships are the new norm
2:30
Michael
Brosnahan
New insights into the behavior of Alexandrium fundyense life cycle
stages through imaging flow cytometry
2:45
Florian Koch
Expansion of brown tide blooms caused by Aureombra lagunensis to the
east coast of the United States
3:00
Reagan
Errera
A day in the life of Karenia brevis: Diel variation in cell volume and
ladder-frame polyether production
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October 2013
SYMPOSIUM AGENDA
11:30 H. Kenneth
Hudnell
MONDAY, OCTOBER 28, 2013 (CONTINUED)
3:15-3:45
3:45-4:45
presenter
Break
Speed talKS SeSSion 1
Chair, leanne flewelling
title
HABS IN A CHANGING WORLD Avery Tatters
High CO2 promotes the production of Paralytic Shellfish
Poisoning toxins by Alexandrium catenella from Southern
California waters
SYMPOSIUM AGENDA
BLOOM DYNAMICS AND ECOLOGY Mindy Richlen
Population structure and genetic dynamics of toxic Alexandrium
fundyense (Dinophyceae) blooms in coastal embayments
poster
number
C-3
B-11
Sugandha Shankar
Role of ammonium in Alexandrium fundyense blooms in the Gulf B-16
of Maine and Georges Bank
Jennifer Vreeland
Estuarine influence on the phytoplankton community structure
during a Karenia brevis bloom on the Florida Gulf coast
B-25
Bill Richardson
Use of dissolved inorganic and organic phosphorus by the toxic
dinoflagellates Karenia brevis and Karenia mikimotoi
(Dinophyceae)
B-6
Matthew Garrett
Stratification influences on the migratory behavior of the harmful B-3
alga Karenia brevis
Mengmeng Tong
The food quality and dissolved inorganic nutrient effect on the
B-23
growth and toxicity of Dinophysis acuminata from North America
Marci Savage
Nutrient and bloom dynamics before dredging of Lake
Neatahwanta, a shallow, freshwater lake in upstate New York
B-13
Christopher Main
The Vibrio-HAB connection: Investigating the influence of iron
on formation of Vibrio biofilms and ROS production by
Heterosigma akashiwo
B-5
TOxICITY AND PATHOGENICITY Dalton Steele
Purification and characterization of indole alkaloids from a central t-16
Florida isolate of the freshwater cyanobacterium, Fischerella sp.,
as inhibitors of vertebrate development in the zebrafish
(Danio rerio) embryo model
Ryan Cassell
Brevetoxin photoaffinity probes for the identification of the
native brevetoxin receptor
t-1
Jan Landsberg
The application of immuno-histochemistry as a tool for
investigating suspected, historic, or unrecognized HAB-related
animal mortality and disease events
t-10
4:45-5:00
Break
5:00-6:30
poster Session 1: HABs in a Changing World; Bloom Dynamics
and Ecology; Toxicity and Pathogenicity
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TUESDAY, OCTOBER 29, 2013
ORALS SESSION 4: PREVENTION, CONTROL, AND MITIGATION
Chair, Marc Suddleson
Time
Presenter
8:30
KevinSellner Optionsinmitigatingcyanobacteriablooms
8:45
Kaytee
Inhibitionofthedinoflagellatecellcycleafterinoculationwiththe
Pokrzywinski algicidalcompoundIRI-160AAexcretedbyShewanellasp.IRI-160
9:00
MarkWarner Examiningtheresponseofharmfuldinoflagellatestothebacterial
algicideIRI-160AA:Differencesingrowth,physiologyandmodeofaction
acrosslaboratoryandfieldcollectedsamples
9:15
Meredith
Howard
Untanglingtheeffectsofanthropogenicversusnaturalnutrientsources
andimplicationsforharmfulalgalbloomsintheSouthernCalifornia
Bight
9:30
Richard
Bartleson
HighCaloosahatcheeRiverandestuarynutrientloadingsandone
harmfulalgalbloomafteranother
9:45-10:15
10:15-11:15
Title
Break
Poster Session 2:BloomDynamicsandEcology;HumanDimensionsof
HABs;MonitoringandManagement
ORALS SESSION 5: MONITORING AND MANAGEMENT - PART 1
Chair, Rob Magnien
EricaSeubert Implementationofsolidphaseadsorptiontoxintracking(SPATT)forthe
monitoringharmfulalgalbloomsinsouthernCalifornia
11:30
Gregory
Doucette
DetectingparalyticshellfishtoxinsusingtheEnvironmentalSample
Processor(ESP):Assaydevelopment&fielddeployment
11:45
Timothy
Otten
AnewframeworkforpublichealthmonitoringofCHABimpacted
drinkingwaterreservoirs:Incorporationofcyanobacterialphysiology
andgenotypesuccessionrates
12:00
NiclasEngene Taxonomicdescriptionsofnovelcyanobacterialbiodiversityfor
monitoring,surveying,andcontrollingcyanobacterialharmfulalgal
blooms(cyanohabs)
12:15
Kathryn
Coyne
12:30-2:00
Effectsofgrowthphase,dielcycleandmacronutrientlimitationonthe
quantificationofHeterosigmaakashiwousingqPCRandSHA
Lunch
ORALS SESSION 6: TOXICITY AND PATHOGENICITY
Chair, Kathi Lefebvre
2:00
2:15
JonathanDeedsGlobalperspectiveontheimpactsofthemarinetoxinpalytoxin
2:30
Alison
Robertson
Linkingfishtohumanillness:confirmationofciguaterafishpoisoning
casesfromSt.ThomasU.S.VirginIslands2010-2012
2:45
JohnBerry
IndolealkaloidsfromFisherellasp.isolatedfromFloridafreshwater
sourcesaretoxictoaquaticvertebratesandinvertebrates
3:00
AshaJaja-
Chimedza
Inhibitionofvertebratedevelopmentinzebrafishembryos(Danio rerio)
exposedtopolymethoxy-1-alkenesisolatedfromfreshwater
cyanobacteria
WayneLitaker Whenhungry,Kareniabrevisgoestoxic
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October 2013
SYMPOSIUM AGENDA
11:15
TUESDAY, OCTOBER 29 (CONTINUED)
3:15-3:45
Break
3:45-4:45
presenter
title
Chair, Barbara Kirkpatrick
BLOOM PREDICTION, FORECASTING, AND MODELING Kristen Thyng
Physical mechanism for Karenia brevis bloom initiation in Texas
p-8
Mary Christman
A satellite-derived predictive model of red tide severity for the
West Florida Shelf
p-2
Allison Allen
NOAA’s Ecological Forecasting Roadmap
p-1
HUMAN DIMENSIONS OF HABS Karen Kavanaugh
From social media to national weather service products: exploring new outreach tools for NOAA’s Harmful Algal Bloom
Operational Forecast System
SYMPOSIUM AGENDA
poster
number
hd-10
Gary Hitchcock
CHANS: Environmental factors influencing Karenia brevis blooms hd-4
on the Florida west coast: Interpretation in support of health
and socio-economic impacts
Vincent Lovko
CHANS: Development of a hazard measurement system for
Florida red tide
hd-5
Katrin Rudge
CHANS: The benefits of participating in broader impacts: A high
school teacher’s perspective
hd-7
Andrew Reich
CHANS: Incidence of Neurotoxic Shellfish Poisonings during
hd-6
Florida red tide (Karenia brevis) blooms: Is shellfish management
effective at mitigating outbreaks?
Perceptions of seafood consumption during Florida red tide
hd-9
blooms: A conjoint analysis
Margaret Byrne
FISHERIES AND FOODWEBS Sheila O’Dea
Florida’s first shellfish harvest closure due to domoic acid
f-2
Christopher Loeffler
Survey of Caribbean ciguatoxins in fish commonly consumed
in St. Thomas, U.S. Virgin Islands
f-1
Justine Schmidt
Variations in the microcystin content of different fish species
collected from a eutrophic lake
f-4
4:45-5:00
Break
5:00-6:30
poster Session 3: Bloom Dynamics and Ecology; Bloom Prediction,
Forecasting, and Modeling; Human Dimensions of HABs; Fisheries
and Foodwebs
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WEDNESDAY, OCTOBER 30, 2013
oralS SeSSion 7: haB genomiCS and other ‘omiCS
time
presenter
Chair, diane greenfield
title
8:30 Allen Place
Some assembly required: Multidomain PKS/NRPS genes in
dinoflagellates
8:45 Julie Kubanek
Metabolomic analysis of harmful algal bloom allelopathy: Deciphering
sublethal impacts on competing phytoplankton
9:00 Kevin Meyer
What exactly are you growing? An assessment of the bacterial
community in commonly used cultures of Karenia brevis and other toxic
and non-toxic dinoflagellates
9:15 Frances
Van Dolah
Global analysis of mRNA half-lives and de novo transcription in the
Florida red tide dinoflagellate, Karenia brevis
9:30 Morgan
Steffen
A systems biology approach to understanding Microcystis blooms
9:45-10:15
Break
oralS SeSSion 8: human and animal health
10:15 Heather
Barron
Chair, Jan landSBerg
Hematologic and biochemical parameters in sea birds with
brevetoxicosis in southwest Florida
10:30 Kathi Lefebvre Common effects of chronic domoic acid exposure in zebrafish, sea lions,
mice and humans
10:45 John Ramsdell Ciguatoxin and Hawaiian monk seals: monitoring blood toxin levels
in northwestern Hawaiian Islands seals and evaluation of foraging under
stress using a mouse model
Ciguatera fish poisoning in St. Thomas, U.S.V.I.: Exposure, symptoms,
and recovery
11:15-11:30
Break
11:30-12:30
poster Session 4: Bloom Dynamics and Ecology; HAB Genomics and
Other ‘Omics; Human and Animal Health; Toxicity and Pathogenicity
12:30-2:00
Lunch
oralS SeSSion 9: monitoring and management - part 2
Chair, vinCent lovKo
2:00 Gregory Boyer MERHAB-LGL: Monitoring and event response in the Lower Great
Lakes– 10 years hence
2:15 Corinne Gibble Widespread detection of the freshwater toxin microcystin at the
land-sea interface within Monterey Bay, CA
2:30 Neil
Harrington
The Jamestown S’klallam tribe responds to an emerging threat to human
health: Diarrhetic Shellfish Poisoning in Washington state
2:45-4:45
haB Community town hall meeting
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October 2013
SYMPOSIUM AGENDA
11:00 Lynn Grattan
THURSDAY, OCTOBER 31, 2013
oralS SeSSion 10: Bloom prediCtion, foreCaSting, and modeling Chair, dennis mcgillicuddy
time
presenter
title
8:30 Barbara Hickey Pacific Northwest Toxins Project
8:45 Clarissa
Anderson
Forecasting the terrestrial influence on domoic acid production:
a mechanistic approach
9:00 Richard Stumpf Climatological analysis of blooms in Lake Erie and seasonal forecast
9:15 David Millie
Great Lakes ‘Big Data’ and HAB-informatics: using artificial intelligence
to deconvolve Microcystis-environmental relationships
9:30 Darren
Henrichs
Predicted origins of Karenia brevis bloom formation along the coast of
Texas using an individual-based model
9:45-10:15
10:15 – 11:15
Break
presenter
Chair, alina Corcoran
title
SYMPOSIUM AGENDA
HAB GENOMICS AND OTHER ‘OMICS Kathleen Pitz
Saxitoxin gene structure and representation in non-toxic and
toxic Alexandrium tamarense species
poster
number
o-10
Lisa Campbell
Cellular response to osmotic stress in Karenia brevis
o-4
David Jayroe
Stress response in Karenia brevis: Changes in ribosomal RNA
o-7
G. Jason Smith
Transcriptome analysis of the diatom Pseudo-nitzschia
australis reveals pathways associated with domoic acid
biosynthesis
o-12
Holly Bowers
Assessing pre-bloom, sub-surface populations of Pseudonitzschia in the San Pedro shelf region of southern California
o-2
MONITORING AND MANAGEMENT
Marco Hatch
Real-time detection of harmful algae at a tribal marine
aquaculture site
m-9
Brian Gregson
Real-time water quality monitoring for HAB events using
autonomous portable water laboratories and data
visualization “cloud” analysis
m-2
Michelle Tomlinson
Quantifying cyanobacteria and high biomass blooms from
satellite
m-8
PREVENTION, CONTROL, AND MITIGATION Katherine Perri
An investigation into the effects of shoreline use on
cyanobacterial abundance in Sodus Bay, Lake Ontario
Cm-6
Wenjun Jiang Photocatalytic degradation of microcystin-LR by Rose Bengal
Cm-3
Helena Pound
Isolation of bacterial capable of degrading microcystin-LR
Cm-7
Kevin Owen
Proposed harmful algal bloom control technologies: Floating
desalination and water pumping plants
Cm-5
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THURSDAY, OCTOBER 31, 2013 (CONTINUED)
11:15-11:30
Break
11:30-12:30
poster Session 5: HAB Genomics and Other ‘Omics; Monitoring and
Management; Prevention, Control, and Mitigation
12:30-2:00
Lunch
oralS SeSSion 11: Bloom dynamiCS and eCology - part 2
Chair, gregory doucette
2:00 Stephanie
Moore
Alexandrium bloom ecology in Puget Sound: Cyst dynamics,
growth, transport, and climate pathways
2:15 Dennis
McGillicuddy
Georges Bank: A leaky incubator of Alexandrium fundyense blooms
2:30 John Ryan
Planktonic layer phenomena in the ecology of toxigenic Pseudonitzschia: examples from coastal California
2:45 Katherine
Hubbard
Spatial and temporal variability of toxic Pseudo-nitzschia spp. in the
Gulf of Maine during summer 2012
3:00 Susanne
MendenDeuer
Linking individual cell behaviors with the population distribution of the
raphidophyte Heterosigma akashiwo
3:15-3:45
Break oralS SeSSion 12: fiSherieS and foodWeBS
Chair, richard pierce
Justin Liefer
Ecological and environmental factors affecting Caribbean ciguatoxins
and their trophic transfer to reef dwelling fishes
4:00
Jennifer
Phillips
Using genetic markers to determine the effect of seeding on the
distribution of a saxitoxin-resistant mutation in Mya arenaria
4:15
Emily Smith
Evaluation of potential oyster contamination from cyanobacterial toxins
4:30-4:45
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October 2013
Closing remarks
SYMPOSIUM AGENDA
3:45
POSTER
SCHEDULE
POSTER SCHEDULE
S
POSTER SESSION 1
MONDAY, OCTOBER 28, 2013 5:00 – 6:30 PM
presenter
title
poster number
(*speed talk)
HABS IN A CHANGING WORLD Lora Fleming
Linking human health and well being with weather, climate and the
environment with harmful algal blooms (HABs)
C-1
Harold Flores
Quintana
Development and evaluation of a direct LC-MS/MS method for
determination of DMSP in biological samples
C-2
Avery Tatters
High CO2 promotes the production of Paralytic Shellfish Poisoning
toxins by Alexandrium catenella from Southern California waters
C-3*
BLOOM DYNAMICS AND ECOLOGY Matthew Garrett Stratification influences on the migratory behavior of the harmful
alga Karenia brevis
B-3*
B-5*
Bill Richardson
Use of dissolved inorganic and organic phosphorus by the toxic
dinoflagellates Karenia brevis and Karenia mikimotoi (Dinophyceae)
B-6*
Bill Richardson
Use of dissolved inorganic and organic nitrogen by the toxic
dinoflagellates Karenia brevis and Karenia mikimotoi (Dinophyceae)
B-7
Mindy Richlen
Population structure and genetic dynamics of toxic Alexandrium
fundyense (Dinophyceae) blooms in coastal embayments
B-11*
Marci Savage
Nutrient and bloom dynamics before dredging of Lake,
Neatahwanta a shallow, freshwater lake in upstate New York
B-13*
Sugandha Shankar Role of ammonium in Alexandrium fundyense blooms in the Gulf
of Maine and Georges Bank
B-16*
Charles Tilney
Comparing the diel vertical migrations of sympatric Karlodinium
veneficum (Dinophyceae) and Chattonella subsalsa
(Raphidophyceae) in laboratory columns
B-21
Mengmeng Tong
The food quality and dissolved inorganic nutrient effect on the
growth and toxicity of Dinophysis acuminata from North America
Estuarine influence on the phytoplankton community structure
during a Karenia brevis bloom on the Florida Gulf coast
B-23*
Jennifer Vreeland
TOxICITY AND PATHOGENICITY Ryan Cassell
Brevetoxin photoaffinity probes for the identification of the native
brevetoxin receptor
Wei Chen
Localization of brevetoxins and photoaffinity labeling of target
proteins
t-1*
t-2
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October 2013
B-25*
ORAL
POSTER
PRESENTATIONS
SCHEDULE
Christopher Main The Vibrio-HAB connection: Investigating the influence of iron on
formation of Vibrio biofilms and ROS production by Heterosigma
akashiwo
ORAL
POSTER
PRESENTATIONS
SCHEDULE
POSTER SESSION 1 (C0NTINUED)
MONDAY, OCTOBER 28, 2013 5:00 – 6:30 PM
presenter
title
poster number
(*speed talk)
Courtney Cocilova
Brevetoxin metabolism and physiology – a freshwater model
of morbidity in endangered sea turtles
t-3
Allan Goodman
Structure activity relationship of brevenal derivatives
t-5
I-Shuo Huang
Geitlerinema sp. produces a novel ichthyotoxic cyanobacterial
toxin
t-7
Asha JajaChimedza
Bioassay-guided purification of a bioactive carotenoid from
the freshwater cyanobacterium, Cylindrospermopsis raciborskii
t-8
Jan Landsberg
The application of immuno-histochemistry as a tool for investigating t-10*
suspected, historic, or unrecognized HAB-related animal mortality
and disease events
Christina Lydon
Identification of cyanobacterial toxins involved in the apparent
intoxication of dolphins in the Florida Keys
t-11
Sean O’Mara
Are fatty acid amides responsible for fish mortality during
Prymnesium parvum blooms?
t-12
Carlton
Rauschenberg
Quantification of domoic acid and okadaic acid in shellfish and
algae using an LC/MS/MS (triple quadrupole) approach
t-13
Dalton Steele
Purification and characterization of indole alkaloids from a central
Florida isolate of the freshwater cyanobacterium, Fischerella sp.,
as inhibitors of vertebrate development in the zebrafish
(Danio rerio) embryo model
t-16*
Bingxue Zheng
Development of capillary electrophoresis as part of a micro-total
analytical system for microcystins
t-17
POSTER SESSION 2
TUESDAY, OCTOBER 29, 2013 10:15 – 11:15 AM
presenter
title
BLOOM DYNAMICS AND ECOLOGY Deana Erdner
Natural mortality during the decline of Alexandrium blooms
poster number
(*speed talk)
B-10
K. David Hambright An experimental assessment of the importance of propagule
pressure and community resistance on the invasion success of
the toxigenic haptophyte Prymnesium parvum
B-12
Sarah Holmes
B-14
Exploring the possibility of cheating between strains of harmful
algal bloom-forming Microcystis aeruginosa
POSTER SESSION 2 (CONTINUED)
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POSTER SESSION 2 (C0NTINUED)
TUESDAY, OCTOBER 29, 2013 10:15 – 11:15 AM
presenter
title
poster number
(*speed talk)
Jayme Smith
The influence of nutrient and light perturbations on natural
phytoplankton assemblages containing the toxin producing diatom
Pseudo-nitzschia
B-19
William
Stringfellow
Unprecedented Microcystis bloom in northern California’s
Bay-Delta Estuary and impact on zooplankton abundance
B-22
Kathryn Van
Alstyne
Environmental factors affecting the release of dopamine and
oxidants by the bloom-forming green seaweed Ulvaria obscura
B-26
HUMAN DIMENSIONS OF HABS Nick Boase
Thinking alike? Comparing expert and public perceptions of the
benefits and risks of consuming shellfish
hd-1
A modified conceptual model integrating health and environmental hd-2
impact assessment: Harmful Algal Blooms
Lora Fleming
CHANS: Public perception of fertilizer ordinances and Florida
red tides
hd-3
Porter Hoagland
Linking Florida red tides to human health effects: Data issues and
preliminary signals
hd-11
Kate Kohler
Where and how do individuals receive and prefer to receive
information on Florida red tide?
hd-12
Zongchao Li
CHANS: Framing the red tide story: Daily newspaper coverage in
southwest Florida
hd-8
MONITORING AND MANAGEMENT Ann Abraham
LC-MS methodology for biomarkers of brevetoxins in the eastern
oyster (Crassostrea virginica) and hard clam (Mercenaria sp.)
exposed to Karenia brevis blooms
m-1
Alan Hails
Successful ocean observatories at work on a harmful algal bloom
on the West Florida Shelf
m-3
Sherwood Hall
Facilitating implementation of the receptor binding assay for PSP
m-4
Sandra Shumway
Biofouling tunicates on aquaculture gear as potential vectors of
harmful algal introductions
m-5
Samantha Weber Monitoring nutrients in Sodus Bay New York
m-6
Ben Whitenack
m-10
Assessing the strengths and weaknesses of each Florida HAB
reporting system
m-11
October 2013
Page 21
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Lawrence Younan Determination of phytoplankton groups using Turner Designs’
PhytoFind
ORAL
POSTER
PRESENTATIONS
SCHEDULE
Lora Fleming
POSTER SESSION 3
TUESDAY, OCTOBER 29, 2013 5:00 – 6:30 PM
presenter
title
ORAL
POSTER
PRESENTATIONS
SCHEDULE
BLOOM DYNAMICS AND ECOLOGY Natalie Dou
A persistent bloom of Anadyomene J.V. Lamouroux
(Anadyomenaceae, Chlorophyta) in Biscayne Bay, Florida
poster number
(*speed talk)
B-1
Amanda Ellsworth
Monitoring strategies for the ciguatera-causing benthic
dinoflagellate, Gambierdiscus
B-2
Gabriela Hannach
Seasonal patterns in nutrients and phytoplankton in Quartermaster
Harbor, an enclosed Puget Sound embayment with recurrent
harmful algal blooms
B-4
Yoonja Kang
Effects of nutrients and grazing mortality on the abundance of
Aureoumbra lagunensis during a Florida brown tide bloom in 2012
B-15
Raphael Kudela
Stimulation of domoic acid production from transient changes
in nutrients
B-17
Alex Leynse
Gambierdiscus nutrient uptake kinetics
B-20
Laura Markley
Potential HAB species found in the Caloosahatchee Estuary, Florida
B-24
Regina Radan
Growth and toxin production of the mixotrophic dinoflagellate
Alexandrium catenella
B-27
BLOOM PREDICTION, FORECASTING, AND MODELING Allison Allen
NOAA’s Ecological Forecasting Roadmap
p-1*
Mary Christman
A satellite-derived predictive model of red tide severity for the
West Florida Shelf
p-2*
Edward Davis
Assessment of the eastern Gulf of Mexico Harmful Algal Bloom
Operational Forecast System: A comparative analysis of forecast
skill and utilization, 2004-2012
p-3
Katherine Derner
Assessment of the western Gulf of Mexico Harmful Algal Bloom
Operational Forecast System: Analysis of forecasts and utilization
over the first two operational years, 2010-2012.
p-4
Jason Lenes
Silicon control of diatom competitors against the red tide
dinoflagellate, Karenia brevis, within the eastern Gulf of Mexico
p-5
Susan Lubetkin
Habitat modeling of Pseudo-nitzschia distribution and toxicity in
the coastal waters of the northwest Pacific using non-parametric
multiplicative regression
p-6
Grace Maze
Time series analysis for periods with high concentration versus
periods with low concentration of Karenia brevis
p-7
Kristen Thyng
Physical mechanism for Karenia brevis bloom initiation in Texas
p-8*
Page 22
TUESDAY, OCTOBER 29, 2013 5:00 – 6:30 PM
presenter
title
poster number
(*speed talk)
FISHERIES AND FOODWEBS Christopher
Survey of Caribbean ciguatoxins in fish commonly consumed in
Loeffler
St. Thomas, U.S. Virgin Islands
f-1*
Sheila O’Dea
Florida’s first shellfish harvest closure due to domoic acid
f-2*
Richard Pierce
Bioaccumulation of brevetoxins and major metabolites in filterfeeding and carnivorous mollusks exposed to natural Karenia brevis
harmful algal blooms
f-3
Justine Schmidt
Variations in the microcystin content of different fish species
collected from a eutrophic lake
f-4*
HUMAN DIMENSIONS OF HABS Margaret Byrne
Perceptions of seafood consumption during Florida red tide blooms: hd-9*
A conjoint analysis
Gary Hitchcock
CHANS: Environmental factors influencing Karenia brevis blooms on hd-4*
the Florida west coast: Interpretation in support of health and
socio-economic impacts
Karen Kavanaugh From social media to national weather service products: Exploring hd-10*
new outreach tools for NOAA’s Harmful Algal Bloom Operational
Forecast System
CHANS: Development of a hazard measurement system for Florida
red tide
hd-5*
Andrew Reich
CHANS: Incidence of Neurotoxic Shellfish Poisonings during Florida
red tide (Karenia brevis) blooms: Is shellfish management effective
at mitigating outbreaks?
hd-6*
Katrin Rudge
CHANS: The benefits of participating in broader impacts: A high
school teacher’s perspective
hd-7*
POSTER SESSION 4
WEDNESDAY, OCTOBER 30, 2013 11:30 AM – 12:30 PM
presenter
title
poster number
(*speed talk)
BLOOM DYNAMICS AND ECOLOGY L. Kellie Dixon
Karenia sp. in SW Florida – trajectories of nitrogen, silica, other
water quality, and taxonomic components during three
extended blooms
Page 23
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October 2013
B-8
ORAL
POSTER
PRESENTATIONS
SCHEDULE
Vincent Lovko
presenter
title
poster number
(*speed talk)
Alexis Fischer
The effect of temperature conditioning on Alexandrium fundyense
cyst germination dynamics in a shallow estuarine system
B-18
Ari Nissanka
Responses of a natural Karenia bloom and other taxa of the West
Florida Shelf to single nutrient, multiple nutrient, or estuarine water
additions
B-9
ORAL
POSTER
PRESENTATIONS
SCHEDULE
HAB GENOMICS AND OTHER ‘OMICS Scott Anglin
Characterizing sense and antisense RNA populations in Karenia
brevis at different times of the diel cycle
o-1
Dianne Greenfield
Elucidating the genetic diversity of Microcystis aeruginosa within
a toxic bloom in South Carolina coastal ponds using
454-pyrosequencing technology
o-3
Grant Jones
Involvement of multiple eIF4ES in mRNA recruitment in
dinoflagellates
o-5
Darcie Ryan
Karenia brevis reference transcripome assembly and transmembrane o-9
protein sequence discovery
Pengfei Sun
Epoxide hydrolases in Karenia brevis: Key enzymes in brevetoxin
biosynthesis
o-11
April Woods
Role of ROS signaling in the domoic acid biosynthetic pathway
o-13
HUMAN AND ANIMAL HEALTH Preston Kendrick Effects of chronic low-dose domoic acid exposure on mitochondrial
function in mice
h-1
Anne Rolton
The effects of field exposure of Crassostrea virginica to Karenia brevis: h-2
Histopathology and brevetoxin accumulation in gametes
Cathy Walsh
Immune function in rescued manatees exposed to brevetoxins
TOxICITY AND PATHOGENICITY Andrea Bourdelais Ladder frame polyethers as potential drug transporters
h-3
t-18
Elizabeth Elliott
Uptake mechanism of fluorescent conjugates of Karenia brevis
ladder frame polyether compounds
t-19
Nick Fowler
A chemical analysis of Karenia papilionacea
T-4
Meghan Grandal
Determining functional effects of brevetoxin and brevetoxin
antagonists
t-6
Page 24
presenter
title
poster number
(*speed talk)
Tanya Hogue
Development of an LCMS assay for determination of amino acid
concentration in Karenia brevis
t-9
Jen McCall
Development of competitive fluorescence-based binding assays for
brevetoxins and brevenal
t-14
Nina Neill
Localization of brevisin-daunorubicin conjugate in SJCRH30 cells
t-20
Susan Niven
Isolation and characterization of a brevenal epimer from cultured
Karenia brevis
t-15
POSTER SESSION 5
THURSDAY, OCTOBER 31, 2013 11:30 AM – 12:30 PM
presenter
title
poster number
(*speed talk)
HAB GENOMICS AND OTHER ‘OMICS Assessing pre-bloom, sub-surface populations of Pseudo-nitzschia
in the San Pedro shelf region of southern California
o-2*
Lisa Campbell
Cellular response to osmotic stress in Karenia brevis
o-4*
Kelly Fridey
Translational response to heat stress in the Florida red tide
dinoflagellate, Karenia brevis
o-6
David Jayroe
Stress response in Karenia brevis: Changes in ribosomal RNA
o-7*
Megan Meek
Development of PCNA as a molecular biomarker for growth status
of Karenia species in Gulf of Mexico red tides
o-8
Kathleen Pitz
Saxitoxin gene structure and representation in non-toxic and toxic
Alexandrium tamarense species
o-10*
G. Jason Smith
Transcriptome analysis of the diatom Pseudo-nitzschia australis
reveals pathways associated with domoic acid biosynthesis
o-12*
Mark Van Asten
The development and validation of a multiplex qPCR assay for the
detection of toxin producing cyanobacteria
o-14
MONITORING AND MANAGEMENT
Danielle Dupuy
A synoptic view of Florida inland algal blooms
m-7
Brian Gregson
Real-time water quality monitoring for HAB events using
autonomous portable water laboratories and data visualization
“cloud” analysis
m-2*
Marco Hatch
Real-time detection of harmful algae at a tribal marine aquaculture
site
m-9*
Page 25
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October 2013
ORAL
POSTER
PRESENTATIONS
SCHEDULE
Holly Bowers
POSTER SESSION 5
THURSDAY, OCTOBER 31, 2013 11:30 AM – 12:30 PM
presenter
title
poster number
(*speed talk)
Michelle Tomlinson Quantifying cyanobacteria and high biomass blooms from satellite
m-8*
PREVENTION, CONTROL, AND MITIGATION
Anamari Boyes
Innovative drinking water treatment for taste and odor removal
with naturally occurring bacteria found in surface water
Cm-1
Amanda Burson
Mitigation of a toxic Alexandrium bloom using hydrogen peroxide
Cm-2
Wenjun Jiang
Photocatalytic degradation of microcystin-LR by Rose Bengal
Cm-3*
Kevin O’Shea
Photooxidation of domoic acid
Cm-4
Kevin Owen
Proposed harmful algal bloom control technologies: Floating
desalination and water pumping plants
Cm-5*
Katherine Perri
An investigation into the effects of shoreline use on cyanobacterial Cm-6*
abundance in Sodus Bay, Lake Ontario
Helena Pound
Isolation of bacterial capable of degrading microcystin-LR
ORAL
POSTER
PRESENTATIONS
SCHEDULE
Cm-7*
Page 26
DINOFLAGELLATE HAB SCIENCE: THEN AND NOW
Karen a. Steidinger
Florida Fish and Wildlife Conservation Commission
100 Eighth Avenue SE, St. Petersburg, FL 33701, USA
Page 27
S
October 2013
ORAL
KEYNOTE
PRESENTATIONS
ADDRESS
Early dinoflagellate HAB research was geared toward
autecology of the causative organism and was conducted
on a local geographic scale. Breakthroughs in phytoplankton
culturing methods allowed scientists to bring the organisms
and their environments into the laboratory for experimentation
and characterization. Neurotoxins and other bioactive
compounds were suspected and tested for but often not chemically characterized
until years later. Global advances in HAB methodology, instrumentation, technology,
and analyses have advanced the science to the point that a species is not just a Latin
binomial but a string of multi-character traits of morphology, ultrastructure, biochemical
constituents (including pigments, toxins, sterols, etc.), gene sequences, life cycle stages,
ecophysiology, geographical distribution, optical signature, and other defining points in
space-time. Today, scientists still study toxins and their analogs, toxin gene sequences,
stressors that affect toxin production, subtle population differences, metabolic alteration
of structure and potency, and the role of bacteria in toxin synthesis. In the 1970s,
scientists recognized the importance of life cycle stages and the possibility of seed beds
in initiating recurring blooms. Today scientific advancement has resulted in a model
that predicts Gulf of Maine Alexandrium fundyense blooms from population dynamics,
seed beds, and physical forcing. Yet, benthic resting stages are not the only players in
recurring blooms; other life cycle stages such as pellicle cysts and pelagic populations
may represent a continuing source of inoculum. Many HAB models involve physical
forcing at some stage of bloom development. Indeed, the physics of the coastal oceans,
whether it relates to upwelling, tidal fronts, thin layers, or circulation patterns is often
critical to predicting HABs. Coupled biophysical models are constantly being pursued
to better understand the system. Ocean observing systems including satellite sensors
and new tools such as chemical and optical detection devices aboard fixed and mobile
platforms are the future for monitoring and forecasting. However, refinement of these
and new applications requires basic research on the HAB organisms, their biological
consortia, the environment, and feedback loops.
ORAL
PRESENTATIONS
ORAL PRESENTATIONS
S
FORECASTING THE TERRESTRIAL INFLUENCE ON DOMOIC ACID
PRODUCTION: A MECHANISTIC APPROACH
Clarissa R. Anderson1, Christopher A. Edwards1, Nicole L. Goebel1, and Raphael M.
Kudela1
University of California Santa Cruz, 1156 High St., Santa Cruz, CA 95073, USA
1
Page 31
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October 2013
ORAL PRESENTATIONS
Physical-biological models are an increasingly important tool for expanding our
heuristic view of phytoplankton community structure. Ideally, biological models will
have a mechanistic component based on first principles of planktonic life cycles and
physiology. Several studies from the Monterey Bay and southern California suggest that
the dominant HAB-former, the toxigenic diatom genus, Pseudo-nitzschia, is alternately
associated with 1) upwelling pulses 2) periods of river runoff and 3) resuspension of
seeding populations into the euphotic zone during upwelling and storms. Laboratory
manipulations of toxigenic Pseudo-nitzschia species in culture have shown that
production of its deadly neurotoxin, domoic acid (DA), is in turn often a function of silicic
acid or phosphate limitation at different phases of Pseudo-nitzschia growth (Bates et al.
1991; Fehling et al. 2004; Pan et al. 1996). Increased nutrient inputs from river runoff are
expected to reduce water column Si:N ratios on time scales relevant to phytoplankton
growth (Kudela 2008). At the same time, these changes in nutrient stoichiometry
are likely coupled to increased loading of urea and other regenerated forms of N,
which further enhance toxin production by at least some species of Pseudo-nitzschia
(Howard et al. 2007; Kudela et al. 2008). We hypothesize that heavy river discharge
to the Monterey Bay following the first large storms of the rainy season - “first flush”
events - increases the toxicity of Pseudo-nitzschia blooms in response to an adjustment
in nutrient stoichiometry. The altered stoichiometry in favor of Si-limitation and
regenerative forms of N increases particulate DA concentrations due to a physiological
stress response associated with DA production. To examine this, we developed a
mechanistic model of DA production, defined as the product of the phytoplankton
growth rate, DA-producing phytoplankton biomass, and a DA production factor where
the growth rate follows the Michaelis-Menten form for individual nutrient limitation.
Various forms of nutrient limitation (Si-lim, N-lim) are tested as adjustments to the DA
growth factor. Two tunable parameters determine the maximum rate of production
and its rate of decline with increasing growth. Our first-year effort has focused on
optimizing model parameters with bootstrapping methods to fit laboratory data from
Si-limited chemostats. We have also tested these fits against batch culture data from
the literature (Fehling et al. 2004) in order to compare our one-dimensional model with
a recently published mechanistic DA model (Terseleer et al. 2013). In this presentation,
we show model comparison results and discuss the implications of a carbon-based
approach (Terseleer et al. 2013) versus our simpler deterministic model for examining
DA production as a function of nutrient ratios and nitrogen source. An advantage to the
latter approach is its suitability for incorporation into a wide variety of NPZ-Regional
Ocean Model Systems without requiring major modification to the NPZ construct and
thereby facilitating model evaluation against observations of “first flush” events in the
Monterey Bay.
HEMATOLOGIC AND BIOCHEMICAL PARAMETERS IN SEA BIRDS WITH
BREVETOxICOSIS IN SOUTHWEST FLORIDA
Heather W. Barron, DVM, Dipl. ABVP-avian1, Richard D. Bartleson, PhD2, Katherine B.
McInnis, DVM1, Helen L. Ingraham, DVM1 and Carolyn Cray, PhD3
Clinic for the Rehabilitation of Wildlife (CROW), 3883 Sanibel-Captiva Rd., Sanibel, FL
33957, USA
2
Sanibel-Captiva Conservation Foundation Marine Lab, Sanibel, FL, 33957 USA
3
University of Miami Miller School of Medicine, Division of Comparative Pathology, P.O.
Box 016960 R-46, Miami, FL 33101 USA.
ORAL PRESENTATIONS
1
Morbidity and mortality in sea birds from red tide events appears to be on the rise
and thus of immediate concern in conservation medicine. Brevetoxins are algal toxins
produced by the dinoflagellate, Karenia brevis (the Florida red tide organism) and other
algal species. Animals may be exposed by ingestion or inhalation of the toxin. More than
2,000 birds representing over a 150 different species present annually to the wildlife
hospital at CROW. In 2012, 439 birds representing 22 different species presented to
CROW with suspected brevetoxicosis, 44 of which were confirmed via ELISA assay.
These toxins cause neuronal and muscle cell depolarization, leading to neurologic signs
in affected animals including ataxia, loss of palpebral reflex, nystagmus, and tremors.
Other clinical signs documented here at CROW that are not routinely reported in the
avian literature include bradycardia, melena, hematochezia, paralysis of the nictating
membrane, decreased vent tone, coughing, and dyspnea. Clinical signs appear to vary
to some degree across species lines. Brevetoxins are metabolized by the liver and
excreted by the kidneys, potentially leading to disease in these organs. However, there
has been little documentation of the clinicopathologic changes seen in sea birds with
brevetoxicosis, which has made effective treatment problematic. Other rehabilitation
centers report treatment success rates of 33-46%; the release rate at CROW in 2012
was 55%. This study determined the seroprevalence of brevetoxins in a wide variety of
piscivorous and omnivorous birds presented to CROW. Additionally, the clinicopathologic
extent of disease in double crested cormorants was determined by documenting
changes in hematologic and plasma biochemical profiles, thus enabling more effective
treatment. The study also determined the range of clinical signs in a variety of different
species of birds positive for brevetoxins to enhance clinical recognition of the disease
among biologists, veterinarians, and wildlife centers.
Page 32
HIGH CALOOSAHATCHEE RIVER AND ESTUARY NUTRIENT LOADINGS
AND ONE HARMFUL ALGAL BLOOM AFTER ANOTHER
Richard D. Bartleson1, Eric C. Milbrandt1, Mark A. Thompson1
Sanibel-Captiva Conservation Foundation Marine Lab, 900A Tarpon Bay Rd. Sanibel, FL,
33957, USA
1
Page 33
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October 2013
ORAL PRESENTATIONS
Surface runoff has replaced much of the groundwater inflows to the Caloosahatchee
Estuary. This runoff in the Caloosahatchee watershed is high in nutrients and colored
dissolved organic matter. Freshwater flow from part of the watershed and Lake
Okeechobee are regulated by three water control structures. When the water control
structure S79 is closed, blooms of cyanobacteria such as Microcystis and Anabaena
occur in the stagnant, fresh water section of the river to the east. These blooms and
their toxins are sometimes passed downstream when flows through S79 are resumed.
When S79 is open, dissolved nutrients from the watershed and the Lake can be
transmitted far downstream and into the Gulf due to low light penetration through the
dark water. Low densities of submersed plants in the Estuary facilitate the transport
of dissolved nutrients. Often, freshwater discharge rates through S79 are high enough
to send plumes of high nutrient water great distances into the Gulf. Sometimes when
these high discharges are occurring, the offshore phytoplankton populations include
Karenia spp. and/or Trichodesmium erythraeum. The high nutrient loading rate into the
Gulf during these events can support growth of dense populations of Karenia brevis
well away from shore. The nitrogen fixing T. erythraeum, which can provide nutrients to
Karenia and may also produce toxins, can also benefit from the high discharge plumes.
Blooms of a variety of other phytoplankton species, some of which are potential toxin
producers regularly occur at the interface of the Caloosahatchee Estuary and the Gulf
of Mexico. In 2011, blooms of two potential brevetoxin producing species, Fibrocapsa
japonica and Chatonella subsalsa, discolored the water along Sanibel beaches. Blooms
of other potentially toxin producing phytoplankton including Pseudo-nitzschia spp.
frequently occur. Thousands of fish and birds and over 200 manatees died in the estuary
during the red tide of 2012-2013. A filamentous, nitrogen fixing cyanobacteria, Lyngbya
majuscula also occasionally overgrows seagrasses in a large area of the estuary after
Karenia blooms. And other macroalgae also reach high biomass levels after red tides
kill algal grazers. The presence of problematic nitrogen fixing cyanobacterial species
throughout the Caloosahatchee River and Estuary and into the Gulf of Mexico shows the
need to substantially reduce phosphorus loading rates which average over 240 metric
tons yr-1 and range to 500 metric tons yr-1.
INDOLE ALKALOIDS FROM FISHERELLA SP. ISOLATED FROM FLORIDA
FRESHWATER SOURCES ARE TOxIC TO AQUATIC VERTEBRATES AND
INVERTEBRATES
Katherine Walton1, Patrick D. L. Gibbs2, Michael C. Schmale2, Miroslav Gantar3 and John
P. Berry1
Department of Chemistry and Biochemistry, Marine Science Program, Florida
International University, 3000 NE 151st Street, North Miami, FL 33181
2
Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric
Sciences, University of Miami, 4600 Rickenbacker Causeway, Miami, FL 33146
3
Department of Biological Sciences, Florida International University, 11900 SW 8th Ave,
Miami, FL 33199
ORAL PRESENTATIONS
1
Fischerella and related genera in the Stigonemataceae, a family of branched filamentous
cyanobacteria, are widely distributed in freshwater systems in the U.S. and globally.
We used the zebrafish (Danio rerio) embryo as a model of vertebrate development
- specifically as a relevant freshwater species of teleost fish – to identify, purify and
subsequently characterize toxic metabolites from cultures of Fischerella isolated from
inland Florida waters. Using this approach, we identified a series of indole alkaloids,
including previously described hapalindoles, as well as a novel nitrile-containing
congener, as extracellular toxic components in culture medium. Toxicity in the
zebrafish embryo was highly variable with structural variant – as evidenced by a range
of “toxitypes” - and consequently pointed to apparent structure-activity relationships
with respect to developmental toxicity. In addition to the zebrafish embryo, purified
metabolites were evaluated for mosquito larvicidal activity in the Aedes aegypti model,
as a relevant representative of freshwater invertebrate, and, likewise, found to be toxic.
Although indole alkaloids from the Stigonemataceae have been previously identified as
putative allelopathic (i.e. anti-algal, antimicrobial) metabolites from this family, this is
the first report to suggest toxicity to relevant vertebrate and invertebrate species, and
support this family of compounds as cyanoHAB toxins with possible implications for both
human and ecosystem health.
Page 34
MERHAB-LGL: MONITORING AND EVENT RESPONSE IN THE LOWER
GREAT LAKES – 10 YEARS HENCE.
Gregory L. Boyer1, Steven W. Wilhelm2, Joseph Makarewicz3, Mary Watzin4, Joseph
Atkinson5, Richard Becker6, Mohammed Sultan7, Charles O’Neill8, Timothy Mihuc9 and
Sue B. Watson10
State University of New York -CESF, Syracuse NY 13210 USA
University of Tennessee, Knoxville, TN 37996, USA
3
The College at Brockport, Brockport NY 14420, USA
4
North Carolina State University, Raleigh, NC 27695, USA
5
University of Buffalo, Buffalo, NY 14260, USA
6
University of Toledo, Toledo OH 43606, USA
7
Western Michigan University, Kalamazoo, MI, USA
8
New York Sea Grant, Ithaca NY 14853, USA
9
State University of New York at Plattsburgh, Plattsburgh NY 12901, USA
10
Environment Canada, Burlington, ON L7R 4A6 Canada
1
2
Page 35
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October 2013
ORAL PRESENTATIONS
In 2002-2008, NOAA funded the MERHAB-Lower Great Lakes effort to develop a tierbased monitoring system for cyanobacteria toxins in Lakes Erie, Ontario and Champlain.
This effort combined individual working groups on cyanobacteria monitoring, toxin
chemistry, new detection methods including molecular biology, hydrodynamic modeling,
remote sensing and outreach. MERHAB-LGL participated in 65 research cruises
between 2002 and 2008, plus hundreds of weekly, biweekly and spot sampling trips
including the first whole lake synoptic and temporal surveys of Lake Ontario, Lake Erie
and Lake Champlain. More than 60 publications appeared in scientific journals to date;
the project generated 11 M.S. and 5 Ph.D. dissertations, and MERHAB investigators gave
more than 200 presentations at scientific meetings or workshops. The MERHAB-LGL
effort did not die once funding ended in 2008. The group remains active in all three
lakes and the monitoring efforts developed here have transitioned to State monitoring
efforts in both Vermont and New York State. Since 2008- MERHAB-LGL investigators
have participated 20 additional research cruises looking at cyanobacteria and their toxins
in the lower Great Lakes and Lake Champlain, and continue to develop new methods
for monitoring and detecting cyanobacteria toxins. This includes the application of
ferrybox systems specifically for the detection of cyanobacterial blooms, as well as the
deployment of new buoy-based monitoring systems for cyanobacteria. New methods
have been developed for the measurement of cyanobacteria toxins in fish that bring
into question some of the prior reports on movement of this toxin through the food
web. Molecular techniques also continue to expand with application of metagenomics,
proteomics and metabolomic techniques to both field and laboratory cultures. The
highlights of these advances in monitoring for harmful cyanobacterial algal blooms will
be reviewed and new methods currently under development will be discussed.
NEW INSIGHTS INTO THE BEHAVIOR OF ALExAndRIum FundyEnSE LIFE
CYCLE STAGES THROUGH IMAGING FLOW CYTOMETRY
Michael L. Brosnahan1, David K. Ralston1, Heidi M. Sosik1, Robert J. Olson1, and Donald
M. Anderson1
Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA.
ORAL PRESENTATIONS
1
For two consecutive years, we have deployed an in situ imaging flow cytometer, the
Imaging FlowCytobot (IFCB) in Salt Pond, a kettle pond within the Nauset Marsh
system (Cape Cod, MA, USA) to study an annually occurring bloom of A. fundyense.
Salt Pond is an ideal study site because its A. fundyense blooms are retained by an
interaction between the cells’ swimming behavior and the pond’s bathymetry that
minimizes advective losses to the A. fundyense population under most conditions. As
a consequence, bloom development and termination can be assessed directly from
changes in cell abundance. Deployments within the pond have been made possible
through the development of a cable-free mooring platform that provides power and
communication links to the IFCB. During the first IFCB deployment in 2012, a concerted
transition by A. fundyense to the sexual phase of its life cycle was recorded. This
transition coincided with increasing stratification in the water column and a rapid
increase in the abundance of an Amoebophrya sp. In 2013, the mooring platform was
updated with an automated winch that was used to record profiles of conductivity,
temperature, depth, chlorophyll fluorescence and photosynthetically active radiation
at 15 – 20 minute intervals over the full course of the bloom. The IFCB itself was also
modified so that it could sample from two different depths throughout the deployment.
Data from 2013 will be used to describe the timing and water column location of several
different life cycle stages including vegetative cells, singlet gametes, fusing gametes and
planozygotes.
Page 36
EFFECTS OF GROWTH PHASE, DIEL CYCLE AND MACRONUTRIENT
LIMITATION ON THE QUANTIFICATION OF HETEROSIGmA AKASHIWO
USING QPCR AND SHA
Kathryn J. Coyne1*, Christopher R. Main1, Cameron Doll2, Colleen Bianco1 and Dianne I. Greenfield2
University of Delaware College of Earth, Ocean, and Environment, 700 Pilottown Rd., Lewes, DE 19958;
2
Belle W. Baruch Institute for Marine & Coastal Sciences, 331 Fort Johnson Road, University of South Carolina, Charleston, SC 29412
1
Page 37
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October 2013
ORAL PRESENTATIONS
Quantitative real-time PCR (qPCR) and sandwich hybridization assay (SHA) are two molecular approaches that have been developed for detection and quantification of HAB species. These methods differ in that SHA detects ribosomal RNA (rRNA) from an unpurified and unamplified sample through oligonucleotide hybridization followed by colorimetric detection, whereas qPCR requires isolation of DNA followed by amplification using species-specific primers and probes and fluorometric detection of PCR products. Our overall goals for this project, funded by NOAA MERHAB, are to rigorously compare these methods for enumeration of harmful algal species, using Heterosigma akashiwo as a model alga, and to provide recommendations for the application of these methods in HAB research and management activities. Here, we examined the effects of physiological status on quantification of H. akashiwo by qPCR and SHA. Samples were collected for analysis at several points during the growth phase, over a diel cycle, and from cultures subjected to either nitrogen or phosphorus limitation. Since the same number of cells was collected for each time point or treatment, the results are indicative of the relative content of DNA (for qPCR) or rRNA (for SHA) per cell. During the growth phase experiment, there was a significant increase in both qPCR and SHA signal for cells collected during log phase growth compared to stationary phase, as would be expected for cells with higher growth rates. Over a diel cycle, however, the results for qPCR analysis indicated an increase in DNA content during the light phase, while rRNA content, as measured by SHA, decreased during the light phase. The decrease in rRNA was unexpected, but may be a consequence of managing resources within the cell. Heterosigma akashiwo has a very large nuclear genome, suggesting that cellular resources such as phosphate may be diverted to dNTP synthesis during the light phase for DNA replication. Under macronutrient limitation, only N-stressed cultures exhibited significantly lower nucleic acid content compared to control cultures, resulting in a lower signal for both qPCR and SHA compared to control cultures. Biochemical and photochemical analyses also confirmed higher levels of stress in the N-stressed cultures compared to P-stressed cultures, which were not significantly different from controls. Overall, results of this investigation suggest that growth stage, nutrient stress and diel changes in the cell cycle can have a significant impact on molecular methods for quantitative assessment of HAB species, and that resource managers should take these factors into consideration when evaluating results from SHA and qPCR analysis. GLOBAL PERSPECTIVE ON THE IMPACTS OF THE MARINE TOxIN PALYTOxIN
Jonathan R Deeds1
US FDA Center for Food Safety and Applied Nutrition, 5100 Paint Branch Parkway,
College Park, MD 20740, USA
1
ORAL PRESENTATIONS
The ancient Hawaiian legend of the “Limu Make o Hana” (deadly seaweed of Hana)
recounts the tale of a mythical creature with a shark’s mouth on its back that is burned
and its ashes cast into a tidal pool after it terrorizes a village in Muolea, in the district
of Hana, on the island of Maui. As the legend goes, after this episode the Limu in
the pool became toxic and the pool itself became kapu (taboo) to the Hawaiians who
believed that an ill fate would befall anyone who disturbed the sacred pool. In efforts to
determine the cause of ciguatera fish poisoning, researchers at the Hawaii Institute of
Marine Biology set out to determine the location of this fabled pool where warriors were
once said to smear the limu on spear points to make their wounds fatal. On December
31, 1961 the pool was finally visited and found to contain a previously undescribed
species of coelenterate zoanthid (colonial anemone), later named Palythoa toxica, from
which palytoxin (PLTX) would eventually be isolated. Coincidentally, a fire destroyed the
main building of the Hawaii Marine Laboratory on Oahu that very afternoon.
From its original discovery, PLTX was realized to be one of the most potent non-protein
toxins ever described. While its toxicity is greatly reduced through oral consumption,
fatal poisonings from contaminated seafood have been documented. More recently,
it was discovered that Ostreopsis spp. dinoflagellates also produce PLTX and PLTX-like
compounds. Furthermore, the frequency and intensity of Ostreopsis blooms appears to
be increasing in temperate coastal waters around the world. Blooms of Ostreopsis have
now been linked to inhalational PLTX exposure, so called “Algal Syndrome”, in portions
of the Mediterranean and Adriatic seas. Lastly, for many years it was believed that the
most highly toxic species of coelenterate zoanthids, such as P. toxica, were limited in
distribution to remote locations such as the tide pools on Maui. It is now known that a
highly toxic variety of zoanthid is commonly available in the home aquarium trade and
has been responsible for serious illnesses due to dermal and inhalational PLTX exposure
in maintainers of marine aquaria world-wide. In addition, less toxic, but highly abundant,
PLTX-containing zoanthid varieties are also known to occur throughout the Pacific and
Caribbean regions and have been linked to food-web accumulation of PLTX and PLTX-like
compounds.
PLTX is rare among marine toxins in that it poses risks through multiple routes of
exposure (oral, inhalational, and dermal), and multiple vectors (planktivorous and
carnivorous fish, crustaceans, shellfish, and waters containing Ostreopsis spp. or toxic
zoanthids). All of this makes the list of potential biological effects after exposure to
PLTXs extensive. The occurrence, chemistry, pharmacology, toxicity, and methods of
analysis for PLTXs have all been recently reviewed. Despite this global attention, much
remains to be learned about the biological mechanisms involved in palytoxicosis, a
serious, potentially life threatening, and sometimes lethal syndrome from exposure to
these compounds. This information is critical for the proper assessment of risks and the
determination of potential management strategies to reduce human exposure to this
unique group of marine toxins.
Page 38
DETECTING PARALYTIC SHELLFISH TOxINS USING THE ENVIRONMENTAL
SAMPLE PROCESSOR (ESP): ASSAY DEVELOPMENT & FIELD
DEPLOYMENT
Jinkeng Asong1, Christina M. Mikulski1, Juliette L. Smith2, Bruce A. Keafer2, Roman
Marin III3 Katrina Campbell4, Christopher T. Elliott4, Christopher A. Scholin3, Donald M.
Anderson2, and Gregory J. Doucette1
NOAA/National Ocean Service, Marine Biotoxins Prog., Charleston, SC 29412, USA
Woods Hole Oceanographic Institution, Biology Dept., Woods Hole, MA 02543 USA
3
Monterey Bay Aquarium Research Institute, Moss Landing, CA 95039, USA
4
Queen’s University Belfast, Inst. for Global Food Security, Belfast BT9 5BN UK
1
2
Reference
D.M. Anderson, G.J. Doucette, G. Kirkpatrick, C.A. Scholin , J. Paul, V. L. Trainer, L.
Campbell, R.M. Kudela, R.P. Stumpf, J.R. Morrison. 2012. Harmful algal bloom (HAB)
sensors in ocean observing systems. IOOS Summit 2012 (http://www.iooc.us/summit/).
Community White Paper.
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October 2013
ORAL PRESENTATIONS
The emergence of ocean observing systems (OOS) capable of transmitting real or
near-real time information from remote locations has the potential to transform the
monitoring and management of harmful algal blooms (HABs) and their adverse public
health and socioeconomic impacts (Anderson et al. 2012). Achieving such a benefit
from OOS will require the development of autonomous, in-water sensors able to
detect individual HAB species and the toxins they produce. The Environmental Sample
Processor (ESP) is a robotic, subsurface platform that can monitor concentrations of both
algal cells and toxins, and provide these data to ship- or shore-based facilities in nearreal time. Here we report on the development of a sensor for paralytic shellfish toxins
(PST) to complement the existing ability of the ESP to detect toxigenic Alexandrium spp.
The PST sensor employs a membrane-based, protein array analogous to that developed
previously for detection of domoic acid on the ESP and, similarly, adopts a competitive
ELISA format. The PST sensor is unique in that two toxin-protein conjugates (one
based on saxitoxin (STX), the other on neosaxitoxin (NEO)) are spotted on the array
and a cocktail of two antibodies is used. One of the antibodies was generated against
gonyautoxin (GTX) 2/3 and the other against NEO. Our aim was to conduct two assays
simultaneously on a single array, one sensitive to the N-1 H toxin group (e.g., STX,
GTX2/3, GTX5, C1/2) and the other sensitive to the N-1 OH group (e.g., NEO, GTX1/4,
C3/4), thereby enhancing our ability to accurately quantify diverse PST profiles
associated with various Alexandrium spp. Currently (modifications are ongoing), both
assays show an IC50 value (analyte concentration yielding half-maximal response) in the
low to mid nanomolar range and can accurately quantify certain complex toxin profiles
generated using both mixed reference standards and Alexandrium culture extracts. We
will report on the performance of the PST sensor in its latest configuration and also
provide preliminary data on the first field deployment of an ESP with saxitoxin detection
capability during spring/summer 2013 in the Gulf of Maine.
TAxONOMIC DESCRIPTIONS OF NOVEL CYANOBACTERIAL BIODIVERSITY
FOR MONITORING, SURVEYING, AND CONTROLLING CYANOBACTERIAL
HARMFUL ALGAL BLOOMS (CYANOHABS)
Niclas Engene1, Sarath P. Gunasekera1 and Valerie J. Paul1
Smithsonian Marine Station at Fort Pierce, 701 Seaway Dr., Fort Pierce, FL 34949,USA
1
ORAL PRESENTATIONS
Cyanobacteria of the genus Lyngbya seasonally form extensive blooms in Southern
Florida. The prolific production of bioactive secondary metabolites of many of
these bloom-forming Lyngbya can be hazardous for humans as well as the natural
environment. In our efforts to provide taxonomic clarity, we show phylogenetically
that several of the most prevalent Lyngbya specimens in Southern Florida, in fact,
represent novel cyanobacterial genera. Here we characterize and compare the ecology,
morphology, evolutionary history, and secondary metabolism of these new taxonomic
groups. These taxonomic descriptions provide classification systems that are able to
identify the cyanobacteria and predict their production of toxins and other bioactive
secondary metabolites. These classification systems are important for monitoring,
predicting, or possibly controlling potentially harmful cyanobacterial blooms in Southern
Florida.
Page 40
A DAY IN THE LIFE OF KAREnIA bREvIS: DIEL VARIATION IN CELL
VOLUME AND LADDER-FRAME POLYETHER PRODUCTION
Reagan M. Errera1, Susan Niven2, Andrea Bourdelais2, Lisa Campbell1
Texas A&M University, 3146 TAMU, College Station, TX, 77843, USA
University of North Carolina-Wilmington, 5600 Marvin K. Moss Lane, Wilmington,
NC 28409
1
2
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October 2013
ORAL PRESENTATIONS
The major harmful alga in the Gulf of Mexico, Karenia brevis, produces a suite of ladderframe polyethers (LFP), which include toxic (brevetoxins) and nontoxic (brevenal and
brevisin) compounds. Light intensity and day length have been shown to be important
environmental factors influencing K. brevis growth rate; however, there has been no
examination of the effect of diel cycles and exponential growth period on the production
of LFPs and cell volume. It has been suggested that variation in LFP production among
clones of K. brevis can be clarified through normalizing LFP concentration to cell volume.
Due to the plasticity of K. brevis theca, daily and growth curve volume changes need to
be explored to verify this hypothesis. Differences in cell size, total parent brevetoxins
and brevenal production were evaluated among four K. brevis clones over a diel cycle in
the middle of exponential growth and over exponential growth. Cell volume for all four
clones varied over the diel cycle. Under a 12:12 (light:dark) photoperiod, all four clones
significantly increased cell volume over the light period and decreased cell volume over
the dark period, while no significant change in LFP production was detected. During
exponential growth, three of the four clones decreased cell volume over the 9-day
period, while the fourth clone increased cell volume, again LFP concentrations (pg cell-1)
remained constant. Furthermore, there was no correlation between cell volume and LFP
production among any of the clones. This suggests LFP production is not related to cell
volume and should not be expressed as a concentration of cell volume. Examination of
field populations of K. brevis over several months also indicated a change in cell size over
the course of the bloom, i.e. from initial state to bloom termination.
Differences in cell size, total parent brevetoxins and brevenal production were also
evaluated under four different photoperiods (6:18, 12:12, 18:6 and 24:0; light:dark)
under an irradiance level of 70 μEin m-2s-1. In both the 12:12 and 24:0 photoperiods,
LFP concentrations (pg cell-1) remained constant in all four clones during exponential
growth. As photoperiod increased, LFP production increased in all four clones, with the
highest concentrations of LFP (pg cell-1) observed under a 24:0 photoperiod. Differences
in cell volume during different photoperiods will also be discussed. To determine if
increases in light intensity resulted in an increase in LFP production growth rates, total
parent brevetoxin and brevenal content of K. brevis were also evaluated at an irradiance
level of 140 μEin m-2s-1 for all four clones. Growth rates significantly increased for the
four clones at the higher irradiance, however, LFP concentrations (pg cell-1) decreased.
The increase of LFP production with increased photoperiod but not with light intensity
suggests a connection between diel cycle and LFP production, specifically LFP production
maybe dependent on the light period.
WIDESPREAD DETECTION OF THE FRESHWATER TOxIN MICROCYSTIN
AT THE LAND-SEA INTERFACE WITHIN MONTEREY BAY, CA
Corinne M. Gibble1 and Raphael M. Kudela1
ORAL PRESENTATIONS
1
University of California, Santa Cruz, 1156 High Street, Santa Cruz, CA 95064
Harmful algae have a worldwide distribution and can form extensive blooms with
toxin production in freshwater, estuarine, and marine habitats. Microcystis aeruginosa
blooms and associated toxin microcystin (MCY) are a regular occurrence in freshwater
systems throughout California, but until recently have not been detected in marine
environments. To investigate potential land-sea transfer of this toxin, 28 sites in and
around Monterey Bay were surveyed for evidence of MCY toxin (2010-2011). In years
two and three (2011-2013) 4 major watersheds in the Monterey Bay area were surveyed
for MCY abundance, nutrients, temperature, and alkalinity to identify potential factors
that might be influencing the abundance of MCY at the land-sea interface. In year
one, MCY was detected in 11 of 28 sites. Data from years two and three analyzed by
multiple regression indicated that coastal nutrient loading (nitrate, phosphate silicate,
ammonium, urea) temperature, and time of year were statistically significant predictors
of the amount of MCY toxin in the environment. At some sites we found evidence of
unexpected seasonality. As anticipated, we predominantly saw large increases of MCY in
the autumn; however at some locations we also recorded very high levels of toxin during
spring. Because this toxin has the ability to biomagnify and persist within food webs,
elevated levels within the watershed may increase potential for illness and death of
wildlife and humans in both terrestrial and marine waters. The widespread occurrence
of MCY at low to moderate levels throughout the year and throughout the watershed
demonstrates the potential difficulty of mitigating these impacts.
Page 42
A NEW SUITE OF NUTRIENT KINETIC RELATIONSHIPS FOR
PICOPLANKTON BLOOMS: NON-TRADITIONAL RELATIONSHIPS ARE THE
NEW NORM
Pat Glibert1
University of Maryland Center for Environmental Science, Horn Point Laboratory, PO Box
775, Cambridge MD 21613, USA
Many coastal and estuarine systems are impacted by EDABs (Ecosystem Disruptive Algal
Blooms)- blooms that are not necessarily toxic but which have negative impacts on
ecosystem function. Many EDAB species are of the picoplankton size range; for example,
Synechococcus blooms in Florida Bay and brown tide blooms in coastal lagoons.
Knowledge of nutritional sources supporting such blooms has greatly advanced in the
past few years. It has been shown, for example, that many picoplankton EDAB species
prefer organic forms of nitrogen (DON) and/or chemically reduced N forms (NH4 or
urea) over oxidized forms (NO3) of nitrogen. This pattern ahs been substantiated by
measurements of uptake rates and also by the finding that many picoplankton have high
proportions of NH4 transporters but few, if any, NO3 transporters. This is in contrast to
diatoms that have many copies of NO3 transporters. Moreover, for some cyanobacteria
the high affinity transporter for NH4 is not regulated at all, being constitutively
expressed.
These findings underscore that these species are able to thrive under conditions
of elevated NH4, urea and organic N. The non-traditional kinetic relationships have
important implications for parameterizing these rates in developing ecosystem models;
standard Michaelis Menten kinetics would not be appropriate to apply in these
conditions. Developing ecosystem models for Florida Bay are beginning to incorporate
these non-traditional kinetic relationships.
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October 2013
ORAL PRESENTATIONS
Here we show the nitrogen uptake kinetic relationships for picocyanobacteriallydominated blooms from Florida Bay. These blooms have been a common feature in
this system for many years, although from year to year have varied in intensity (but
not in dominant species). The kinetic relationships for NO3, NH4 and urea did not
follow classic saturation responses in most cases. The typical response for NO3 was a
linear relationship for uptake as a function of increasing concentration. This pattern
is consistent with a lack of NO3 regulation at the level of transport across the cell
membrane, and is also consistent with diffusion od substrate into the cell. For NH4 and
urea there were two common responses. The first was inhibition of uptake as a function
of increasing concentration. This pattern is suggestive of down-regulation of nutrient
transport so as to maintain a constant flux of nutrient into the cells. The second common
pattern was biphasic, with both suppression of uptake at intermediate concentration
levels, followed by a secondary increase at much higher concentrations. This pattern
suggests that the cells regulated uptake up to a point, but that a diffusional effect
occurred at the highest concentrations measured.
THE ROLE OF COASTAL OCEAN ACIDIFICATION IN PROMOTING AND
ExACERBATING THE EFFECTS OF HARMFUL ALGAL BLOOMS
Christopher J. Gobler1, Theresa K. Hattenrath-Lehmann1, and Ryan B. Wallace1
School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook, NY
11794-5000, USA
ORAL PRESENTATIONS
1
During the past decade, oceanographers have become acutely aware of the decline
in ocean pH associated with the loading of carbon dioxide (CO₂) predominantly from
the combustion of fossil fuels. Concurrently, it has become increasingly recognized
that coastal habitats within which harmful algal blooms occur are already experiencing
acidification, often of a magnitude far exceeding the ‘worst case scenario’ conditions
predicted for the open ocean later this century (>1,000 ppm CO2). Excessive nutrient
loading to coastal waters often promotes algal blooms, including HABs, and the demise
of these algal blooms typically yields high rates of microbial respiration that lowers
dissolved oxygen concentrations. A second, often overlooked consequence of microbial
respiration is the production of CO2 and a reduction in seawater pH. Since many HABs
often occur following the decline of other algal blooms they are more likely to develop in
acidified environments that have elevated levels of pCO2. This presentation will highlight
the interactions and feedbacks between nutrient loading, HABs, and acidification and
how they affect marine organisms and ecosystems. Novel laboratory and ecosystem
observations regarding the ability of acidification to significantly increase (p<0.05) the
growth and toxicity of North Atlantic isolates and blooms of Alexandrium fundyense will
be presented. Data regarding the ability of ecosystem disruptive algal blooms including
brown tides caused by Aureococcus anophagefferens and Aureoumbra lagunensis, to
manipulate the pH, oxygen, and pCO2 levels within estuaries will be presented. Finally,
data demonstrating that the development of HABs within high CO2 ecosystems cause
harm to marine animals beyond the effects posed by the elevated CO2 or the harmful
algae alone will also be presented.
Page 44
CIGUATERA FISH POISONING IN ST. THOMAS, USVI: ExPOSURE,
SYMPTOMS, AND RECOVERY
Lynn M. Grattan,1 Sparkle M. Roberts,1 Alison Robertson,2 Elizabeth Radke,3 Margaret
Abbott,3 and J. Glenn Morris3
University of Maryland School of Medicine, 110 S. Paca Street Street, Baltimore, MD
21201
2
Gulf Coast Seafood Laboratory, U.S. Food and Drug Administration, 1 Iberville Drive,
Dauphin Island, AL 36528
3
University of Florida, P.O. Box 100009, 2055 Mowry Road, Gainesville, FL 32610
1
Ciguatera fish poisoning (CFP), a harmful algal bloom syndrome, is the most common,
non-bacterial foodborne illness associated with fish consumption in the United States
and globally. While the syndrome has been recognized for more than six centuries, more
than 50 different symptoms have been associated with the symptom complex with
varying levels of consistency. Moreover, patterns of recovery may range from 24 hours
to several years. In contemporary thought, syndrome variations are potentially explained
by geographic toxin diversity, therefore region-specific studies are indicated.Toward this
end, we examined a series of patients who presented to the emergency department at
the Roy Schneider Hospital and were diagnosed with CFP.
Meanwhile, it is concluded tht CFP remains a problem in St. Thomas, USVI, with Red
Hind, Barracuda, and Red Snapper being the most risky fish. Neurologic and dermal
symptoms are the most persistent symptoms and full recovery may be expected within
the first year for most people. Ongoing outreach and education are essential to reducing
the risk of exposure and illness.
Page 45
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October 2013
ORAL PRESENTATIONS
Exposure was assessed by type and quantity of fish consumed as well as analysis of
meal remnant samples. Sixty-two study participants were examined within five days of
emergency room discharge, with measures to capture potential symptoms or symptom
complaints. This included symptom checklists and standard measures of cognition,
mood, and sensory perception. Follow-up evaluations were completed three months and
one year post exposure. Preliminary findings indicate no seasonal variation of CFP cases.
At least twelve different types of finfish were associated with illness, with the most
frequently reported fish consumed being Red Hind (16%), Barracuda (13%), and Red
Snapper (13%). Most of the fish associated with illness were purchased from or gifted by
local fishers. Where meal remnants were available and analyzed (n=20), 80% were found
to be ciguatoxic. The most frequently reported clinical symptoms were diarrhea; upper
and lower extremity weakness and tingling; and abdominal pain and nausea. Sensory
and neuropsychological screening indicated no abnormalities. With respect to chronicity,
56% of patients presenting to the emergency department with CFP had dermal and/or
neurologic symptoms three months later and 13% continued to have unusual and vague
neurologic complaints one year post exposure. Finally, 56% of patients had at least one
previous episode of CFP. Data collection and anlysis are ongoing.
THE JAMESTOWN S’KLALLAM TRIBE RESPONDS TO AN EMERGING
THREAT TO HUMAN HEALTH: DIARRHETIC SHELLFISH POISONING IN
WASHINGTON STATE
Neil Harrington1, Vera L. Trainer2, Leslie Moore2, Brian D. Bill2, Jerry Borchert3 and
Bich-Thuy L. Eberhart2
Jamestown S’Klallam Tribe, 1033 Old Blyn Highway, Sequim, WA 98382
Marine Biotoxins Program, Environmental Conservation Division, Northwest
Fisheries Science Center, National Marine Fisheries Service, National Oceanic and
Atmospheric Administration, 2725 Montlake Blvd. E, Seattle, WA 98112
3
Office of Shellfish and Water Protection, Washington State Department of
Health, 111 Israel Rd SE, Tumwater, WA 98504
1
ORAL PRESENTATIONS
2
Three people contracted diarrhetic shellfish poisoning (DSP) after eating mussels
collected at Sequim Bay State Park on the Olympic Peninsula of Washington
State in June 2011. These were the first confirmed cases of DSP in the United
States. The emergence of this new threat to public health had an immediate
impact on the Jamestown S’Klallam Tribe whose subsistence and commercial
shellfish beds were located nearby. In 2012 a collaborative research project
on DSP in Sequim Bay and Washington State by NOAA, the Tribe and the
Washington State Department of Health (WDOH) looked at the presence of
Dinophysis species and concentrations of diarrhetic shellfish toxins (DSTs) in
shellfish and phytoplankton. In an effort to provide advanced warning of shellfish
toxicity using rapid screening methods for toxins, analysis of extracts using both
antibody- and activity-based screening assays were compared to the standard
regulatory method, liquid chromatography-mass spectrometry. Weekly sampling
illustrated that D. acuminata was the primary species present during toxic events
and increases in cell density usually predicted increases in shellfish toxicity. In
addition, it was found that dinophysistoxin-1 (DTX-1) was the primary responsible
toxin isomer in WA and mussels generally concentrated the toxin more than
other species of shellfish. Greater than average snowpack and freshwater runoff
in both 2011 and 2012 in the northwest may have contributed to conditions
favorable for dinoflagellate blooms. These findings are being used to better
utilize monitoring data from the SoundToxins and Olympic Region Harmful
Algal Blooms (ORHAB) phytoplankton monitoring groups to prioritize shellfish
collection and analyses for DSTs to protect public health.
Page 46
NUTRIENTS SUPPORTING KAREnIA bREvIS BLOOMS: RESULTS OF THE
ECOHAB: KAREnIA PROGRAM
Cynthia Heil1, Deborah Bronk2, L. Kellie Dixon3, Gary Hitchcock4, Gary Kirkpatrick3,
Margaret Mulholland5, Judith O’Neil6, John Walsh7, Robert Weisberg7, Matthew Garrett8,
Jason Lenes7, Rachel Sipler2, Bill Richardson8, Lynn Killberg-Thorsen2, Kevin Meyer6, Leo
Procise5 and Breanne Walsh6
Bigelow Laboratory for Ocean Sciences, East Boothbay, ME 04544, USA
Department of Physical Sciences, The College of William & Mary, Virginia Institute of
Marine Science, Gloucester Point, VA, 23062, USA
3
Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236, USA
4
RSMAS/MBF, University of Miami, Miami, FL 33149, USA
5
Department of Ocean, Earth and Atmospheric Sciences, Old Dominion University,
Norfolk, VA 23529, USA
6
Horn Point Laboratory, Univ. of Maryland Center for Environmental Science, Cambridge,
MD 21613, USA
7
College of Marine Science, University of South Florida, St. Petersburg, FL 33701, USA
8
Fish and Wildlife Research Institute, Florida Fish and Wildlife Conservation Commission,
St. Petersburg, FL 33701, USA
1
2
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October 2013
ORAL PRESENTATIONS
Blooms of the toxic dinoflagellate Karenia brevis predate coastal eutrophication but
continue to annually plague the eastern Gulf of Mexico with fish kills, marine mammal
mortalities, human respiratory irritation and significant economic losses. The ECOHAB:
Karenia Program was a collaborative, multi-faceted approach to identify, assess and
model the nutrient sources fueling these blooms. Three blooms were sampled, one at
initiation phase (2008), one in a high biomass, nearshore maintenance phase (2007)
and a third in a lower biomass, offshore maintenance phase (2009) as well as a ‘no
bloom’ year (2010). Nutrient sources supporting K. brevis blooms are multiple, diverse
and complex: the largest nitrogen source was Trichodesmium blooms (N2 fixation and
release in com-bination with decay and recycling of biomass), followed by nutrient
release from zooplankton grazing, decay of red tide-related dead fish, benthic nutrient
flux, photochemical nutrient production, nitrification, estuarine inputs and pelagic
N2 fixation. Reduced nitrogen forms dominant the sources, many of which were
individually sufficient to support observed bloom biomass. Project results suggest that
toxins play a significant role in altering nutrient sources and dynamics of larger, high
biomass, fish killing blooms compared with smaller, low biomass blooms by reducing
some nutrient sources (e.g. zooplankton grazing) while enhancing others (e.g. decay
of dead fish). K. brevis cells are physiologically flexible with regards to their nutrient
acquisition strategies, which allow them to respond rapidly to changing environmental
conditions. Cells can take up and grow on a variety of inorganic and organic N and P
sources, grow rapidly (up to1 day-1) under organic enrichment conditions, and possess
unique carbon acquisition strategies. Results suggest that management options for K.
brevis blooms, while limited for nutrient reduction strategies, should include reduction
of controllable nutrient sources through best management practices, continued support
and improvement of short-term now-casting and forecasting abilities and products, and
targeted educational outreach efforts.
PREDICTED ORIGINS OF KAREnIA bREvIS BLOOM FORMATION ALONG
THE COAST OF TExAS USING AN INDIVIDUAL-BASED MODEL
D. W. Henrichs1, R. D. Hetland1, L. Campbell1,2
Department of Oceanography, Texas A&M University, College Station, TX 77843
Department of Biology, Texas A&M University, College Station, TX 77843
1
ORAL PRESENTATIONS
2
Bloom formation in Karenia brevis is not the result of rapid cell division but rather, a
combination of physical and physiological processes. Bloom formation in the eastern
Gulf of Mexico is well-studied but the origin of cells contributing to bloom formation
remains unknown. The western Gulf of Mexico experiences more infrequent blooms of
K. brevis and as a result, the origin of cells contributing to bloom formation has received
little attention. A spatially-explicit, individual-based model (IBM) of K. brevis has been
developed to identify potential source locations of cells that may become physically
concentrated along the Texas coast. The model merges the behavioral model of Liu et
al. (2001) with an individual-based population-genetic model incorporating cell division
and sensitivity to environmental temperature and salinity. Current velocities, water
temperature, and water salinity are obtained from the Gulf of Mexico HYCOM model. To
identify potential source(s) of Karenia blooms, the IBM was run in reverse and cells were
inserted at locations along the coast of Texas by date according to results obtained from
the Imaging FlowCytobot in Port Aransas and count data from Texas Parks and Wildlife
Department. Comparisons between bloom years and non-bloom years suggest blooms
of K. brevis in Texas originate in the southern Gulf of Mexico, near the Bay of Campeche,
an observation supported by recent reports of blooms off the western Yucatan. Prebloom identification of cells offshore of Texas, carried north by currents, would help to
validate the model results. Running the model in forward time and starting cells in the
southern Gulf of Mexico, using forecast currents, coupled with a downwelling index
(based on the mean monthly along-shore component of the wind at the coast of Texas)
will be used to test the forecasting ability of the individual-based model.
Page 48
PACIFIC NORTHWEST TOxINS PROJECT
Barbara Hickey1, Neil Banas2, Michael Foreman3, Raphael Kudela4, Evelyn Lessard1, Parker
MacCready1, Diane Masson3, Richard Thomson3, Thomas Connolly5, Nancy Kachel1,
Kristen Davis6, Isaac Fine3, Susan Geier1, Sarah Giddings1, Susan Lubetkin1, Ryan McCabe1,
and Samantha Siedlecki2
School of Oceanography, University of Washington, Seattle, WA 98195, USA
Joint Institute for the Study of the Atmosphere and Ocean, University of Washington,
Seattle, WA 98195, USA
3
Fisheries and Oceans Canada, Institute of Ocean Sciences, Sidney, BC V8L4B2, Canada
4
Department of Ocean Sciences, University of California, Santa Cruz, CA 95064, USA
5
Woods Hole Oceanographic Institute, Woods Hole, MA 02543, USA
6
Civil and Environmental Engineering and Earth System Science, University of California,
Irvine, CA 92697, USA
1
2
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October 2013
ORAL PRESENTATIONS
The goal of PNWTOX is to determine transport routes for toxigenic Pseudo-nitzschia
in the Pacific Northwest. The role of the freshwater plume from the Columbia River in
bloom initiation, nutrient provision, and transport is a central focus of the project. The
study includes data analysis and model development based on an extensive suite of
biological/chemical/physical observations acquired on ten 21 d cruises over a 4-year
period, from satellites, and from multiple deployments of moored arrays and surface
drifters throughout the region. The massive data set, which includes biological growth
and grazing rates, nutrients, and oxygen data, has guided the development of a regional
ROMS transport and ecosystem model. The model has been thoroughly validated and
includes multiple processes, such as Puget Sound and Strait of Georgia/Fraser River
estuarine exchange and remotely forced waves, which our studies have shown to be
critical for adequate representation of upwelling from the slope to the shelf, and for
accurate provision of nutrients to the region. The model captures observed patterns
and magnitudes of chlorophyll, nitrate, bottom oxygen, temperature, Columbia plume
location and particle transport routes on both seasonal and event (several day) scales
with good accuracy. HAB transport routes examined in detail for 2004-2007 reproduced
the observational results showing that toxic cells are delivered to the WA coast not from
a single source, as suggested in prior studies, but from two sources, one north and one
south of the WA beaches where razor clam harvest is impacted by HABs. The existence
of sources both north and south of the affected beaches is dependent on the fact that
regional currents in the Pacific Northwest reverse seasonally, in contrast to the US East
coast currents. The northern source, the Juan de Fuca Eddy region, is important in late
summer and fall, when currents are southward; whereas the southern source, Heceta
Bank, OR, is the source in late winter/spring, when currents are seasonally northward.
The model results also confirm the hypothesis that the Columbia plume at times acts as
a barrier to onshore transport, while at other times it enhances alongshelf transport. As
a result of the PNWTOX research, the Pacific Northwest regional model, which can easily
be applied to other HABs with known source regions, is now also poised to be used in
HAB and hypoxia regional forecasting efforts.
CHANS: FLORIDA RED TIDES AND COASTAL POPULATIONS AS A
COUPLED NATURE-HUMAN SYSTEM
Porter Hoagland1, Barbara Kirkpatrick2,3, Gary Kirkpatrick2, Gary Hitchcock4, Kate Kohler2,
Vince Lovko2, Steve Ullmann5, Andy Reich6, and Lora Fleming3,4,7
Marine Policy Center, Woods Hole Oceanographic Institution, Woods Hole, MA 02543,
USA
2
Mote Marine Laboratory, Sarasota, FL 34236, USA
3
Department of Epidemiology and Public Health, Miller School of Medicine, University of
Miami, Miami, FL 33177 USA
4
Oceans and Human Health Center, Rosenstiel School of Marine and Atmospheric
Sciences, University of Miami, Miami, FL
5
Programs and Center in Health Sector Management and Policy, University of Miami,
Miami, FL 33177 USA
6
Aquatic Toxins Program, Environmental Health, Florida Department of Health,
Tallahassee, FL
7
European Centre for Environment and Human Health, Peninsula College of Medicine
and Dentistry, Truro, Cornwall, UK
ORAL PRESENTATIONS
1
Coupled nature-human (CNH) systems are now the focus of a growing number of interdisciplinary research programs worldwide. As implied by the term “coupled,” these
systems involve interactions between humans and nature, often affecting the dynamic
characteristics of each component. Both natural and social scientists are engaged in
developing a deeper understanding of these dynamics, focusing on the linkages and
feedbacks affecting the trajectories of coupled system behavior. Several researchers
have begun to identify the generic aspects of nature-human couplings. Many of these
aspects have been adapted from the field of ecology, where the dynamic characteristics
of ecological systems have been studied for decades. These aspects include system
heterogeneity, time lags, reciprocal feedbacks, thresholds, surprises, legacies, and
resilience. The presence of such aspects has implications for the stability and persistence
of particular ecosystem states, leading potentially to further implications for human
heath and welfare. This talk reviews a specific type of natural hazard-human coupling
that relates to coastal blooms of toxic marine algae, drawing examples primarily
from human interactions with blooms of the toxic dinoflagellate Karenia brevis from
the eastern Gulf of Mexico. This talk introduces a set of HAB Symposium “speed”
presentations relating to different aspects of an ongoing multi-institutional and interdisciplinary research project that examines Florida red tides as a type of CNH system.
We present examples of the generic aspects of CNH systems in the context of Florida red
tides, and we discuss also some of the challenges involved in compiling relevant data to
support our analytical efforts.
Page 50
UNTANGLING THE EFFECTS OF ANTHROPOGENIC VERSUS NATURAL
NUTRIENT SOURCES AND IMPLICATIONS FOR HARMFUL ALGAL BLOOMS
IN THE SOUTHERN CALIFORNIA BIGHT
Meredith D.A. Howard1, Karen McLaughlin1, Martha Sutula1, David A. Caron2, Yi Chao3,4,
Hartmut Frenzel4, Alyssa Gellene2, Kendra Hayashi5, Burton Jones6 , Raphael M. Kudela5,
Michael J. Mengel7, Nikolay Nezlin1, George Robertson7, Ashmita Senqupta1 and Erica
Seubert2
Southern California Coastal Water Research Project, 3535 Harbor Blvd., Costa Mesa, CA
92626 USA
2
University of Southern California, 3616 Trousdale Parkway, Los Angeles, California
90089-0371, USA
3
University of California, Los Angeles, 405 Hilgard Ave, Los Angeles, CA
4
Remote Sensing Solutions, Inc., 2824 East Foothill Blvd., Pasadena, California 91107
5
6
King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of
Saudi Arabia
7
Orange County Sanitation District, 10844 Ellis Avenue, Fountain Valley, CA
92708Fountain Valley, CA 92728
1
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ORAL PRESENTATIONS
Eutrophication of coastal waters is a global environmental issue, with demonstrated
links between anthropogenic nutrient inputs and increased frequency and occurrence of
harmful algal blooms. However, in upwelling-dominated ecosystems, such as southern
California, untangling the relative influence of natural versus anthropogenic nutrient
sources on coastal waters has proved to be more complex. In these ecosystems, there
has been a perception that anthropogenic nutrient inputs are small relative to upwelling,
and thus, have little effect on nearshore productivity. However, recent studies in the
Southern California Bight (SCB) have provided evidence to the contrary. A one year
study in 2010 determined that anthropogenic nitrogen loads from wastewater effluent
discharged to ocean outfalls were equivalent to modeled upwelling nitrogen loads in
five of six sub-regions located proximal to the coastline. Furthermore, stable isotope
analysis of nitrate, ammonia, and particulate organic matter showed that nitrogen
from wastewater effluent comprised up to half of the total nitrogen in phytoplankton
and zooplankton located proximal to an ocean outfall in the fall 2012. Similar isotopic
analysis is underway for samples collected during a toxic Pseudo-nitzschia bloom in
the same location in spring 2013 and will be presented. These findings suggest that
anthropogenic nutrients, mainly wastewater effluent, can provide a significant source of
nitrogen for algal bloom development in southern California coastal waters.
SPATIAL AND TEMPORAL VARIABILITY OF TOxIC PSEudO-nITZSCHIA
SPP. IN THE GULF OF MAINE DURING SUMMER 2012
Katherine A. Hubbard1,2, Alison Sirois3, Jane Disney4, Leanne Flewelling2, Sheila O’Dea2,
Steve Morton5, Alison Robertson6, Harold A. Flores Quintana6, Judith L. Kleindinst1,
Dennis J. McGillicuddy1, Donald M. Anderson1
Woods Hole Oceanographic Institution, Woods Hole, MA, 02543
Fish and Wildlife Research Institute, 100 8th Ave SE, St. Petersburg, FL 33701
3
Maine Department of Marine Resources, PO Box 8, West Boothbay, ME, 04575
4
Mount Desert Island Biological Laboratory, PO Box 35, Salisbury Cove, ME 04672
5
NOAA/ NOS, Hollings Marine Laboratory, 331 Fort Johnson Rd, Charleston, SC 29412
6
Gulf Coast Seafood Laboratory, U.S. FDA, 1 Iberville Drive, Dauphin Island, AL 36528
1
ORAL PRESENTATIONS
2
Although repeated shellfish harvest closures for the Amnesic Shellfish Poisoning (ASP)
toxin domoic acid (DA) have occurred in eastern Canadian provinces since 1987, the
first ASP closure in New England waters occurred in late July 2012. Prior to the closure,
increases in Pseudo-nitzschia abundance along the coast of eastern ME from Eastport to
Schoodic Peninsula were detected by the Maine Volunteer Phytoplankton Monitoring
Program (Maine Department of Marine Resources). By July 31st, phytoplankton
and shellfish samples from several small embayments in the region tested positive
for DA (with Jellet ASP rapid testing assays), and a closure was issued (area # 64A).
Federal partners at NOAA and the FDA respectively conducted SEM characterization
of Pseudo-nitzschia species and confirmatory toxin testing of shellfish. Emergency
funding provided by NOAA’s CSCOR Harmful Algae Event Response program permitted
additional sampling of nearshore and offshore phytoplankton communities, particulate
DA, and physicochemical parameters at 25 stations during the closure. The highest
cellular abundance (1.3 x 106 cells l-1) occurred in Corea Harbor, and was attributed to
a novel species that is morphologically similar to P. calliantha and genetically similar
to P. pseudodelicatissima/P. cuspidata. Pseudo-nitzschia abundance decreased with
distance from shore, with 104-105 cells l-1 just outside of embayments and 102 -103 cells
l-1 2-3 nautical miles offshore. Pseudo-nitzschia became more abundant west of the
Schoodic region during September and October, and some but not all samples with
high Pseudo-nitzschia abundance tested positive for DA. To assess the effects of species
composition on variability in DA toxicity, a modified automated ribosomal intergenic
spacer analysis (ARISA) was used to identify and quantify Pseudo-nitzschia species in
DNA samples collected during the event, specifically targeting regional species (n=8)
that are morphologically similar to P. pseudodelicatissima. Although DA concentrations
in shellfish during the bloom peak were quickly determined to be below the regulatory
closure limit of 20 ppm and the closure was lifted, the multi-agency response provided
timely, critical information to address the regional threat of ASP. This has led to
enhanced monitoring efforts in ME, including a pilot study to conduct volunteer-based
DNA sampling. As a more extensive regional program for monitoring DA and Pseudonitzschia spp. is adopted, it is important for managers, monitors, and researchers
to continue to learn more about the toxicity, frequency and magnitude of Pseudo-
Page 52
FRESHWATER MANAGEMENT POLICY UPDATE: IMPLEMENTING THE 3RD
PILLAR OF THE CWA, WATERBODY MANAGEMENT, TO FORM A
SYSTEMS APPROACH
H Kenneth Hudnell1 and Wayne Carmichael2
Medora Corporation & UNC-CH, 105 Serrano Way, Chapel Hill, NC 27517, USA
Department of Biological Sciences, Wright State University, Dayton, OH 45435, USA
1
2
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October 2013
ORAL PRESENTATIONS
A systems approach to freshwater management is needed to protect and restore
freshwater in the near term at a cost much less than that currently spent on watershed
management alone. Current US policy fails to protect unimpaired waters or restore
impaired waters. Whereas the EPA estimated in 1972 that 10 to 20 percent of lakes
and reservoirs were eutrophic, the agency now estimates that about 50 percent are
eutrophic or hypereutrophic. A recent EPA assessment of 2008-2009 data for rivers and
streams found that 66 percent contained excessive levels of phosphorus, a primary cause
of eutrophication, up from 47 percent in 2004. The CWA (Clean Water Act) established
three pillars of freshwater protection and restoration, the watershed management
point- and nonpoint-source pollutant-input control programs, and the waterbody
management (Clean Lakes) program, but the latter program was not implemented
to a significant extent. Waterbody management provides the supportive therapy
needed to reduce stress on impaired biochemical processes and enable recovery. Only
full implementation of the CWA provides a systems approach by addressing not only
land-based pollutant inputs, but also atmospheric depositions, existing internal loads,
stressed aquatic biochemical processes, and pollutant-induced impairments. A policy of
watershed management only requires decades to restore designated uses, if ever, and is
excessively expensive. For example, the watershed management plan for Falls Lake, NC,
is estimated to cost about $2 billion by 2035. A systems approach will restore impaired
waterbodies’ designated uses in the near term by directly targeting the impairments,
such as suppressing harmful algal blooms that often produce highly potent toxins
and cause chlorophyll-a, pH, and turbidity impairments. Treatments also can remove
nutrients, inactivate pathogens, degrade toxic substances, and restore the ecological
community balance that historically maintained good water quality. A systems approach
also will lower the cost of restoration and protection. The complementary combination
of within waterbody treatments and pollutant input reductions that stabilize point
source nutrient-input limits (account for only 5-10% of input) and use only the most
effective and cost efficient nonpoint source best management practices would reduce
the overall cost. An estimation of waterbody management cost in Falls Lake is $25
million by 2035. A national multipronged initiative is underway to align policy with
the CWA and establish a systems approach. The initiative includes: 1) improved
Federal legislation; 2) ongoing discussion with Federal agencies; 3) improved state
policy and legislation and; (4) two new nonprofit organizations; the 501(c)(3) Impaired
Waters Restoration Alliance for education and research, and the 501(c)(4) Waterbody
Management Association for advocacy. Significant advances and challenges will be
reported. Audience input will help direct this initiative’s course of action critical for
health, aquatic ecosystems, economies and our nation’s security and wellbeing.
INHIBITION OF VERTEBRATE DEVELOPMENT IN ZEBRAFISH EMBRYOS
(dAnIO RERIO) ExPOSED TO POLYMETHOxY-1-ALKENES ISOLATED
FROM FRESHWATER CYANOBACTERIA
Asha Jaja-Chimedza1, Patrick D. L. Gibbs2, Miroslav Gantar3, and John P. Berry1
Department of Chemistry and Biochemistry, Florida International University, 3000 NE
151st Street, North Miami, FL 33181 USA
2
Science, University of Miami, 4600 Rickenbacker Causeway, 33149, Florida, USA
3
Department of Biological Sciences, Florida International University, 11200 SW 8th Street,
Miami, FL 33199, USA
ORAL PRESENTATIONS
1
Aphanizomenon and Cylindrospermopsis are two toxigenic genera of freshwater
cyanobacteria known to produce a number of toxic metabolites, particularly including
the water-soluble toxins cylindrospermopsin, saxitoxin, and anatoxin-a. Lipophilic
extracts of strains of each of these genera were shown to inhibit the development
of zebrafish (Danio rerio) embryos. Bioassay-guided fractionation using the zebrafish
embryo, as a model of vertebrate development, was subsequently utilized to purify
the toxic chemical components. Chemical characterization of the purified compounds,
including mass spectrometry and NMR (1H, 13C, DEPT, COSY, HMQC, HMBC), revealed the
presence of a homologous series of six polymethoxy-1-alkenes (PMAs). Toxicological data
suggests there is an apparent correlation between chain length and/or methoxylation
of the PMAs and toxicity in the embryos. In addition, exposure of the embryos to
combinations of the PMAs, suggests a possible synergistic effect associated with the
toxicity of these compounds. A culture collection of different strains of cyanobacteria
was screened by LC-MS for the presence of PMAs, where they were identified in a
number of isolates of cyanobacteria, including a Microcystis strain isolated from the
Great Lakes, suggesting that these bioactive compounds may be widespread. Due to the
frequent occurrence of these cyanobacterial genera in freshwater sources, particularly
in association with cyanobacterial harmful algal blooms, they may pose a previously
unrecognized environmental and health risk, including possible bioaccumulation of
these lipophilic toxins in freshwater food-webs.
Page 54
HARMFUL ALGAL BLOOM INTEGRATED OBSERVING SYSTEM FOR THE
GULF OF MExICO
Ann E. Jochens1, Barbara A. Kirkpatrick2, Steven H. Wolfe3
Texas A&M University, 3146 TAMU, College Station, TX 77843, USA
Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236 USA
3
Florida Institute of Oceanography, 830 1st Street S., St. Petersburg, FL 33701, USA
1
2
Page 55
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October 2013
ORAL PRESENTATIONS
Building an integrated observing system for harmful algal blooms in the Gulf of Mexico
is a high priority for the partners that make up the Gulf of Mexico Alliance (GOMA),
led by the five U.S. Gulf States, and the Gulf of Mexico Coastal Ocean Observing
System Regional Association (GCOOS-RA), which is a regional component of the U.S.
Integrated Ocean Observing System that is the U.S. part of the International Global
Ocean Observing System. These two organizations encompass myriad local, state, and
federal agencies, private industry, and academic and research institutions. They have
joined together to plan, design, and implement an integrated, comprehensive, sustained
Harmful Algal Bloom Integrated Observing System (HABIOS) for the Gulf of Mexico.
Under the auspices of the GCOOS-RA and GOMA, a series of three workshops have been
held specifically to develop the HABIOS, and efforts are underway to identify assets and
funding sources to collaboratively build HABIOS as part of the broader Gulf of Mexico
Coastal Ocean Observing System (GCOOS-RA) and Gulf Monitoring Network (GOMA).
The workshops developed a strategy for HABIOS and identified user groups and needs
for HABs data and information. Then, gaps in existing HAB data and products were
identified, and the steps for implementation were explored. The primary beneficiaries
of a fully-functioning HABIOS are shell and fin fisherman (both commercial and
recreational), recreational boaters, and beachgoers, as well as the managers responsible
for protecting public and ecosystem health.
HABIOS is designed to address four focus areas: (a) prediction of bloom initiation,
(b) detection of bloom existence, (c) tracking or monitoring of the bloom, and (d)
forecasting of bloom movement and effects. The fully-developed HABIOS will support
observing of major harmful algal species, with emphasis initially on prominent species,
such as Karenia brevis and Dinophysis spp., that currently are causing shellfish bed
closures, marine life kills, or human illness. It will target not only species that have
surface chlorophyll signals observable by satellite, such as K. brevis, but also species that
do not, such as Pseudo-nitzschia spp. Observing components will include satellite data,
moorings, and gliders or other autonomous profiling instrumentation. The system builds
on existing efforts by state agencies, academics, and federal agencies such as NOAA.
In addition to supporting the broader goals of the GCOOS and GMN, the full HABIOS
will supplement the existing HAB monitoring system by using fixed stations and routine
glider transects to continuously detect specific HAB species and collect data on a suite
of parameters (e.g., nutrients, currents, temperature, salinity, fluorescence, and optical
phytoplankton community structure) that would indicate favorable conditions for HAB
events.
ExPANSION OF BROWN TIDE BLOOMS CAUSED BY AuREOumbRA
LAGunEnSIS TO THE EAST COAST OF THE UNITED STATES
Florian Koch1, Christopher J. Gobler1, Yoonja Kang1, Dianna L. Berry1, Ying Zhong Tang1,
Margaret Lasi2, Linda Walters4, Lauren Hall3, and Jan D. Miller3
Stony Brook University, School of Marine and Atmospheric Sciences, Southampton, NY,
11968
2
St. Johns River Water Management District, Palatka, FL 32177
3
St. Johns River Water Management District, Palm Bay, FL 32909
4
University of Central Florida, Department of Biology, Orlando, Florida 32816
ORAL PRESENTATIONS
1
Brown tides caused by the pelagophyte Aureoumbra lagunensis DeYoe et Stockwell have
formed ecosystem disruptive algal blooms in shallow lagoons of Texas (TX), USA, for
more than two decades but have never been reported elsewhere. During the summer
of 2012, a dense brown tide occurred in the Mosquito Lagoon and northern Indian
River Lagoon along the east coast of Florida (FL), USA. While chlorophyll a levels in this
system have averaged 5 µg L-1 during the past two decades, concentrations during this
brown tide reached ~ 200 µg L-1. Concurrently, levels of nitrate were significantly lower
than average and levels of dissolved organic nitrogen were significantly higher than
average (p<0.001 for both). Sequences of the 18S rRNA gene of the bloom community
and of single cell clonal isolates were identical to those of Aureoumbra lagunensis
DeYoe et Stockwell from TX. The A. lagunensis brown tide in FL bloomed to densities
exceeding 106 cells mL-1 (quantified with a species-specific immuno-label) from July
through September, began to dissipate in October, but maintained densities exceeding
105 cells mL-1 in some regions through December of 2012. The decline of the bloom
was associated with near-hypoxic conditions and more than 30 fish kills reported within
the Mosquito Lagoon in September 2012, a number exceeding all prior reports in this
system dating to 1996. Wild Northern quahog populations (a.k.a. hard clam, Mercenaria
mercenaria) suffered mass die offs during the brown tide and eastern oysters
(Crassostrea virginica) that settled during 2012 were significantly smaller than prior
years. Clearance rates of hard clams and eastern oyster were significantly reduced in
the presence of Mosquito Lagoon bloom water and A. lagunensis monocultures isolated
from the Mosquito Lagoon at densities of ~106 cells L-1. The expansion of harmful brown
tides caused by A. lagunensis to these estuaries represents a new threat to the US
southeast coast. Dynamics of Aureoumbra lagunensis in these lagoons during the 2013
and details of a novel method for quantifying this algae will also be presented.
Page 56
METABOLOMIC ANALYSIS OF HARMFUL ALGAL BLOOM ALLELOPATHY:
DECIPHERING SUBLETHAL IMPACTS ON COMPETING PHYTOPLANKTON
Kelsey Poulson-Ellestad1, Christina Jones2, Jessie Roy1, Facundo M. Fernandez2, Brook
Nunn3, Jon Byrne4, Mark Viant4, Julia Kubanek1,2
Georgia Institute of Technology, School of Biology and Aquatic Chemical Ecology Center,
310 Ferst Drive, Atlanta, GA 30332, USA
2
Georgia Institute of Technology, School of Chemistry & Biochemistry and Aquatic
Chemical Ecology Center, 310 Ferst Drive, Atlanta, GA 30332, USA
3
University of Washington, Department of Genome Sciences, Box 355065, WA 98195,
USA
4
University of Birmingham, School of Biosciences and NERC Metabolomics Facility,
Edgbaston, Birmingham B152TT, UK
1
Page 57
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October 2013
ORAL PRESENTATIONS
How individual species come to be dominant members of marine planktonic
communities is not deeply understood; however, it is thought that chemistry plays a
substantial role. For example, some red tide-forming dinoflagellates produce toxic
compounds that are hypothesized to enhance dinoflagellate fitness by acting as grazer
deterrents, allelopathic agents, or antimicrobial defenses. In field and lab experiments
we have previously shown that the red tide dinoflagellate Karenia brevis is allelopathic,
inhibiting the growth of several co-occurring phytoplankton species, but that K. brevis
compounds other than well-known brevetoxins are responsible for suppressing most
of these species. For phytoplankton competitors, death is a rare outcome of K. brevis
allelopathy: more subtle responses predominate, such as reduced photosynthetic output
and increased cell permeability. We characterized changes in cellular physiology with
NMR and MS-based metabolomics which indicated disruption of primary metabolic
pathways in diatoms exposed to K. brevis. Preliminary proteomic analyses further
supported the finding that metabolic networks are impacted by red tide allelopathy. A
diatom species known to co-occur with K. brevis responded with less dramatic metabolic
alteration than a model diatom whose range does not overlap with K. brevis, suggesting
adaptation among phytoplankton that frequently experience K. brevis blooms. Overall,
systems biology approaches including metabolomics and proteomics offer considerable
opportunities for expanding our understanding of plankton ecology, driving novel
hypotheses about the molecular mechanisms of chemically mediated interactions.
COMMON EFFECTS OF CHRONIC DOMOIC ACID ExPOSURE IN
ZEBRAFISH, SEA LIONS, MICE AND HUMANS
Kathi Lefebvre1, Lynn Grattan2, Elizabeth Frame1, Preston Kendrick1, Sparkle Roberts2,
Emma Hiolski3, Don Smith3, Glen Morris4, and Dave Marcinek5
NOAA Fisheries, NWFSC, Seattle, WA 98112, USA
University of Maryland School of Medicine, Baltimore, MD 21201, USA
3
University of California, Santa Cruz, CA 95064, USA
4
University of Florida, P.O. Box 100009, Gainesville, FL 32610, USA
5
University of Washington, Seattle, WA 98109, USA
1
ORAL PRESENTATIONS
2
It is well known that acute exposure to high levels of the algal toxin domoic acid (DA)
is responsible for a neurotoxic illness known as amnesic shellfish poisoning (ASP)
characterized by vomiting, diarrhea, seizures, memory loss, coma and death. Human
seafood consumers are protected from high level exposure via the monitoring of
seafood and subsequent regulation of harvests based on toxin loads (≥ 20 ppm =
harvest closure). However, there are no protections in place for low-level repetitive
exposure (< 20 ppm = harvest open) because there is a critical knowledge gap regarding
chronic exposure effects. Our research team has investigated the impacts of repetitive
subclinical DA exposure in two laboratory human disease model species (zebrafish and
mice), one naturally exposed sentinel for human health species (California sea lions),
and in “at risk” high seafood consuming humans. Chronic subclinical exposure (below
levels that show overt excitotoxic symptoms) has been shown to significantly alter
gene expression in the CNS, elicit an immune response, and increase toxin sensitivity in
zebrafish. Additionally, the presence of a DA specific antibody was indicated in zebrafish
and validated in naturally exposed sea lions, thereby revealing a potential diagnostic
biomarker for chronic exposure. Mouse and human studies are currently underway
in an effort to verify the utility of the antibody biomarker and relate it to subclinical
neurotoxic effects. A synthesis of results from all models will be presented.
Page 58
ECOLOGICAL AND ENVIRONMENTAL FACTORS AFFECTING CARIBBEAN
CIGUATOxINS AND THEIR TROPHIC TRANSFER TO REEF DWELLING
J.D. Liefer1, A. Robertson1, A.C. Garcia1, T. Smith2, H.A. Flores Quintana1, M. Richlen3, and
D.M. Anderson1
FDA Gulf Coast Seafood Laboratory, 1 Iberville Dr., Dauphin Island, AL 36528, USA
University of the Virgin Islands, #2 John Brewers Bay, St. Thomas, U.S. Virgin Islands
00802, USA
3
Woods Hole Oceanographic Institution, 266 Woods Hole Rd., Woods Hole, MA 02543,
USA
1
2
Page 59
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October 2013
ORAL PRESENTATIONS
Ciguatoxins (CTXs) are cyclic polyether toxins that originate from benthic dinoflagellates
of the genus Gambierdiscus, which often grow in association with common
reef macroalgae such as Dictyota. These toxins undergo biotransformation and
biomagnification in tropical reef food webs. Consumption of CTX-contaminated fish
results in ciguatera fish poisoning (CFP). In the Caribbean, this poisoning syndrome
is characterized by a variety of gastrointestinal and neurological symptoms. The US
Virgin Islands is an area that has sustained high prevalence of CFP that has impacted
human health and important fisheries. To address these impacts, we are examining how
microalgal community composition, reef physical conditions, and water quality affect the
production, biotransformation, and trophic transfer of CTXs. The long-term goal of these
efforts is modeling CTX production dynamics and predicting future outbreaks of CFP in
the greater Caribbean region. Four sites near St. Thomas, US Virgin Islands were sampled
from Jun 2010 to Dec 2012 to assess the production and trophic transfer of Caribbean
CTXs (C-CTX). Epiphytic dinoflagellates were quantified from Dictyota samples collected
monthly to determine species composition and composite C-CTX concentration (which
includes the precursors gambiertoxins) using N2a cytotoxicity assays and LC-MS. In
addition, a variety of fishes representing multiple trophic levels were collected quarterly
from each site and analyzed for C-CTX. Gambierdicsus and C-CTX activity in dinoflagellate
samples were detected year-round at all four sites. The Gambierdiscus C-CTX cell quotas
were highest during colder months, a pattern not previously shown in this region. The
Gambierdiscus cell quota of C-CTX was consistently higher at Black Point, a near shore
site characterized by relatively less mixing and wave action. Herbivorous fish species
also had consistently higher C-CTX concentrations, many above the FDA guidance
threshold of 0.1 ppb, at Black Point compared to the other three sites. These results
indicate the importance of temperature and physical forcing in regulating Gambierdiscus
abundance and the subsequent production and transfer of CTX. We also present an
extensive comparison of C-CTX cell quotas with dinoflagellate species composition and
environmental conditions such as physical hydrography and nutrient concentrations.
WHEN HUNGRY, KAREnIA bREvIS GOES TOxIC
D. Ransom Hardison1,2, William G. Sunda1, Damian Shea2, Patricia A. Tester1, R. Wayne
Litaker1
Center for Coastal Fisheries and Habitat Research, National Centers for Coastal Ocean
Science, National Ocean Service, 101 Pivers Island, Beaufort, NC, 28516, USA.
2
Department of Biology, North Carolina State University, Raleigh, North Carolina, USA
ORAL PRESENTATIONS
1
Karenia brevis is the dominant toxic red tide species in the Gulf of Mexico. It produces
potent neurotoxins (brevetoxins [PbTxs]), which negatively impact human and animal
health, local economies, and ecosystem function. Field measurements have shown that
cellular brevetoxins vary from 1‒68 pg/cell but the source of this variability is uncertain.
The carbon:nutrient balance (CNB) hypothesis, which was originally developed for
terrestrial plants, predicts that nutrient-limited growth will be accompanied by a
diversion of fixed carbon into increased levels of carbon-based defensive compounds or
structures. Consistent with this hypothesis, data from numerous harmful algal bloom
species have shown that nutrient limitation causes an increase in cellular toxin content.
Historically, the effect of nutrients on K. brevis toxicity has been controversial.
This study was undertaken to examine the effect of nitrogen (N) and phosphorus (P)
limitation of growth rate on cellular brevetoxins in diverse K. brevis strains from different
geographic locations. N and P were selected because both have been reported to limit
algal growth in the Gulf of Mexico. N- and P-limited cells had 2- to 3-fold higher PbTx
per unit of cell volume or cell carbon. The percent of total cellular carbon associated
with brevetoxins (%C-PbTx) was 1-4% in N-replete cultures and 5-9% in the N-limited
cells. Similarly, %C-PbTx in P-replete isolates ranged from 0.7 to 2.1% in P-replete cells,
but increased to 1.6‒5% under P-limitation. Because PbTxs are potent anti-grazing
compounds, this increased investment in PbTxs should enhance cellular survival during
periods of nutrient-limited growth. The %C-PbTx for all isolates was also found to
be inversely related to specific growth rate, which is consistent with an evolutionary
tradeoff between carbon investment in PbTxs and other grazing defenses, and carbon
investment in growth and reproduction. In aquatic environments where nutrient supply
and grazing pressure often vary on different temporal and spatial scales, this tradeoff
would be advantageous as it would result in increased net population growth rates. The
variation in PbTx/cell values observed in this study can account for the range of values
observed in the field. These results suggest that N- and P-limitation are important factors
regulating cellular toxicity and adverse impacts during at least some K. brevis blooms.
The relationship between growth and toxicity is being used in models to predict how
the toxicity of K. brevis blooms will change as the bloom densities increase and use up
available nutrients. These models show that the combination of high biomass and high
toxin to biomass ratios that occur when blooms run out of nutrients results in extreme
bloom toxicity, and helps explain why Karenia blooms are often very damaging to
ecosystems, human health, and coastal economies. The results also point to the need
for direct toxin measurements in affected waters to directly assess toxicity and forecast
adverse impacts.
Page 60
GEORGES BANK: A LEAKY INCUBATOR OF ALExAndRIum FundyEnSE
BLOOMS
Dennis J. McGillicuddy, Jr.1
David W. Townsend2
Bruce A. Keafer3
Maura A. Thomas2
Donald M. Anderson3
Department of Applied Ocean Physics and Engineering, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA.
2
School of Marine Sciences, University of Maine, Orono, ME 04469, USA.
3
Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA
1
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October 2013
ORAL PRESENTATIONS
A series of oceanographic surveys on Georges Bank document variability of populations of the toxic dinoflagellate Alexandrium fundyense on time scales ranging from synoptic to seasonal to interannual. Blooms of A. fundyense on Georges Bank can reach concentrations on the order of 104 cells l-1, and are generally bank-wide in extent. Georges Bank populations of A. fundyense appear to be quasi-independent of those in the adjacent coastal Gulf of Maine, insofar as they occupy a hydrographic niche that is colder and saltier than their coastal counterparts. In contrast to coastal populations that rely on abundant resting cysts for bloom initiation, very few cysts are present in the sediments on Georges Bank. Bloom dynamics must therefore be largely controlled by the balance between growth and mortality processes, which are at present largely unknown for this population. Based on correlations between cell abundance and nutrient distributions, ammonium appears to be an important source of nitrogen for A. fundyense blooms on Georges Bank.
LINKING INDIVIDUAL CELL BEHAVIORS WITH THE POPULATION
DISTRIBUTION OF THE RAPHIDOPHYTE HETEROSIGmA AKASHIWO
Susanne Menden-Deuer and Elizabeth L. Harvey
Graduate School of Oceanography, University of Rhode Island, Narragansett, RI
ORAL PRESENTATIONS
In the laboratory, we investigated the effects of biotic and abiotic stimuli on the
individual behaviors and vertical distribution of Heterosigma akashiwo. Using automated
observation and video analysis procedures; we simultaneously investigated the
three-dimensional movements of free-swimming cells and the resultant macroscopic
population distributions of different strains in tanks with a halocline structure. Induced
by both the presence of actively feeding ciliate and dinoflagellate predators and
predator-derived chemical cues, H. akashiwo cells exhibited effective fleeing behaviors.
At the individual cell level, both vertical velocity and swimming direction of the halotolerant HAB alga changed significantly, resulting in population aggregation in low
salinity waters, inaccessible to the less halo-tolerant predator. The ciliate predator
showed no response to algal exudates but the dinoflagellate displayed significant
increases in swimming speed and turning rate when exposed to filtrate from H.
akashiwo. Prey cue–induced changes in predator swimming behavior resulted in an
11% increase in encounter rate and a 25% reduction in the necessary ambient prey
concentration necessary to meet the daily quota of the predator. Our results provide
quantitative evidence for the importance of individual behaviors and chemical cues in
modulating H. akashiwo’s vertical distribution and potential predator pressure, both
factors that are independent of population growth, but critical in predicting abundance,
distribution and bloom formation potential of this HAB alga.
Page 62
WHAT ExACTLY ARE YOU GROWING? AN ASSESSMENT OF THE
BACTERIAL COMMUNITY IN COMMONLY USED CULTURES OF KAREnIA
bREvIS AND OTHER TOxIC AND NON-TOxIC DINOFLAGELLATES
Kevin A. Meyer1, Judy M. O’Neil1, and Byron C. Crump2
University of Maryland Center for Environmental Science: Horn Point Laboratory, 2020
Horns Point Road, Cambridge, MD, 21613, USA
2
Oregon State University, 1500 SW Jefferson Way, Corvallis, OR, 97331, USA
1
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ORAL PRESENTATIONS
Algal cultures are commonly used for experimental studies because of the control
over the system, and organism of interest, it provides the researcher. Harmful algal
bloom forming species are no exception and in some cases, like the Florida Red Tide
dinoflagellate Karenia brevis, were some of the earliest cultures established in the
United States. In many cases the bacterial community is left intact in phytoplankton
cultures (not axenic), which then subjects the bacterial community to the same selective
pressures being used to maintain a single algal species. Bacteria-algal relationships are
well documented but are rarely considered during experiments that use HAB cultures.
Given that some cultures, like the infamous “Wilson clone” were isolated decades ago,
it is likely that bacterial communities in such cultures no longer resemble typical Gulf
of Mexico bacterial communities. The bacterial community was investigated within
seven different isolates of Karenia brevis, an isolate of Karenia mikimotoi, Prorocentrum
minimum, P. rhathymum, Karlodinium veneficum, and Gyrodinium dorsum from Florida
Fish and Wildlife Research Institute and MOTE Marine Laboratory. Genetic analysis was
done using Illumina sequencing and bacterial growth was measured using 3H-Leucine
and Thymidine to better understand the community composition, biological function,
and possible relationships between commonly used cultures and co-cultured bacteria.
GREAT LAKES ‘BIG DATA’ AND HAB-INFORMATICS: USING ARTIFICIAL
INTELLIGENCE TO DECONVOLVE mICROCySTIS-ENVIRONMENTAL
RELATIONSHIPS
David F. Millie1,2, Gary R. Weckman3,1, Gary L. Fahnenstiel2,4, William A. Young II5, Ehsan
Ardjmand3, John A. Fahnenstiel1, Robert A. Shuchman2,4, and Michael J. Sayers2
Palm Island Enviro-Informatics LLC, Sarasota, FL 34232, USA
Michigan Technological University, Michigan Tech Research Institute, Ann Arbor, MI
48105, USA
3
Ohio University, Russ College of Engineering and Technology, Department of Industrial
and Systems Engineering, Athens, OH 45701, USA
4
Michigan Technological University, Great Lakes Research Center, Houghton, MI 49931,
USA
5
Ohio University, College of Business, Management Systems Department, Athens, OH
45701, USA
1
2
ORAL PRESENTATIONS
Nutrient-enriched waters of the Laurentian Great Lakes are plagued by expansive,
recurring blooms of the toxic cyanobacterium, Microcystis aeruginosa. The intensity
of these events have been linked to phosphorus–laden tributary discharges and
concurrent, optimal meteorological conditions. Although such influences appear to
work ‘in tandem’ as controls of HABs, a comprehensive analysis of Microcystis ‘forcing
factors’ has yet to be completed for the Lakes.
Monitoring of the blooms - via invasive sampling, instrumental-based observatories, and
satellite imagery, has resulted in massive, high-dimensional data streams (i.e. ‘Big Data’)
that personnel must search through to identify data structures and relationships. Recent
technological advances have provided for the evolution of computational-intensive
analytics that optimize for the delineation and reproduction of complex patterns within
large databases. Such an ‘informatic’ approach previously has been used by the authors
to denote (apparent) phosphorus threshold concentrations for Microcystis blooms
within Saginaw Bay, Lake Huron from 1990 to 1996 (J. Phycol., 2011; 7: 714–730).
Using diverse databases (imagery, watershed hydrodynamics, meteorology, physical/
chemical limnology), we present a neural network–based approach demonstrating
the selection and quantitation of environmental attributes for modeling Microcystis
abundance dynamics throughout Saginaw Bay (2008-2010) and western Lake Erie (20092011). Model formulations provide interpretable, multi-dimensional response surfaces
for environmental-Microcystis associations, thereby allowing for visualized, qualitative/
quantitative representations of HAB-nutrient relationships and interaction (akin to niche
modeling). Such a heuristic methodology affords a mathematically comprehensive, yet
pragmatic understanding for factors regulating phytoplankton blooms and becomes
a data-driven source upon which to identify ‘target’ nutrient concentrations for
management efforts attempting to reduce/eliminate HABs throughout the Lakes.
Page 64
ALExAndRIum BLOOM ECOLOGY IN PUGET SOUND: CYST DYNAMICS,
GROWTH, TRANSPORT, AND CLIMATE PATHWAYS
S. K. Moore1, B. D. Bill1, L. R. Hay1, K. C. Eldred1, C. L. Greengrove2, J. E. Masura2, N. S.
Banas3, E. P. Salathé4, N. J. Mantua4,5, J. A. Johnstone6, D. M. Anderson7, V. L. Trainer1, and
J. E. Stein1
NOAA, Northwest Fisheries Science Center, Seattle, WA; stephanie.moore@noaa.gov
University of Washington–Tacoma, Tacoma, WA
3
Applied Physics Laboratory, University of Washington, Seattle, WA
4
University of Washington, Climate Impacts Group, Seattle, WA
5
NOAA, Southwest Fisheries Science Center, Santa Cruz, CA
6
University of Washington, Joint Institute for the Study of the Atmosphere and Ocean,
Seattle, WA
7
Woods Hole Oceanographic Institution, Woods Hole, MA
1
2
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ORAL PRESENTATIONS
The Puget Sound Alexandrium Harmful Algal Bloom (PS-AHAB: www.tiny.cc/psahab)
program seeks to understand environmental controls on the benthic (cyst) and
planktonic life stages of the toxic dinoflagellate Alexandrium catenella, and disentangle
the effects of climate pathways on the timing and location of blooms. Spatially detailed
mapping of winter cyst distributions in 2011, 2012, and 2013 found the highest cyst
concentrations in Bellingham Bay in the north and Quartermaster Harbor in central
Puget Sound. However, the viability of cysts at these seed bed areas is low – with fewer
than 54% of cysts germinating when incubated at favorable temperatures. This may
complicate potential relationships between cyst abundances and bloom magnitude
the following season. The time of year that cysts can germinate does not appear to be
determined by endogenous controls. The rate of cyst germination is strongly determined
by temperature. Passive particles released from these seed beds were tracked using a
high-resolution hydrodynamic simulation of Puget Sound and adjacent coastal waters
(MoSSea: http://faculty.washington.edu/pmacc/MoSSea/). In two weeks, particles
released from Bellingham Bay made it out of the Strait of Juan de Fuca to the outer
Washington coast whereas particles released from Quartermaster Harbor mostly stayed
in the main basin of Puget Sound. No particles entered Hood Canal, suggesting that
physical transport mechanisms may prevent toxic cells from contacting shellfish in this
basin of Puget Sound. Laboratory experiments showed that maximal growth rates (~0.30.5 μ d-1) occur over a broad range of temperatures (~14-24°C) at salinities typical for
Puget Sound (20-35 psu). These ranges were used to define favorable habitat for A.
catenella using model output from the MoSSea simulation. A 40-year global climate
projection was regionally downscaled and coupled to MoSSea to determine temporal
and spatial changes to favorable habitat under the A1B greenhouse gas emissions
scenario. A comparison between present-day and circa-2050 conditions allows us
disentangle the effects of three climate pathways on favorable habitat for A. catenella in
Puget Sound: 1) changing ocean inputs (associated with upwelling winds), 2) changing
streamflow magnitude and timing, and 3) increased direct insolation.
A NEW FRAMEWORK FOR PUBLIC HEALTH MONITORING OF CHAB
IMPACTED DRINKING WATER RESERVOIRS: INCORPORATION OF
CYANOBACTERIAL PHYSIOLOGY AND GENOTYPE SUCCESSION RATES
Timothy G. Otten1,2, Theo W. Dreher1 and Hans W. Paerl2
Oregon State University, Department of Microbiology, 220 Nash Hall, Corvallis, OR
97331, USA
2
University of North Carolina at Chapel Hill, Institute of Marine Sciences, 3431 Arendell
St, Morehead City, NC 28557, USA
ORAL PRESENTATIONS
1
High throughput sequencing and quantitative PCR were used to characterize the
spatiotemporal patterns of cyanobacterial harmful algal blooms (CHABs) in two large
river ecosystems - the Klamath River and San Francisco Bay Delta. In recent years both
of these systems have been plagued by recurring, seasonal blooms of toxin-producing
Microcystis spp., as well as diazotrophic genera (e.g., Aphanizomenon and Anabaena)
of relatively unknown toxicity. The sequencing and qPCR data enable us to quantify
population succession rates based on a multilocus analysis of allele frequencies over
time and space. These data, along with detailed physicochemical analyses, can be
used to greatly improve our understanding of how different environmental conditions
may select for toxigenic ecotypes over nontoxic variants. Here we propose a public
health monitoring framework which takes into account laboratory based analyses of
algal physiology (e.g., growth kinetics, cell toxin quotas, stress response) and in situ
observations of population turnover rates from cyanobacterial blooms under natural
settings. The goal of this research is to create and validate a public health sampling
protocol for CHAB impacted reservoirs which minimizes unnecessary costs due to
oversampling, without sacrificing the public health benefits of a proactive drinking water
monitoring program.
Page 66
MANAGING HARMFUL CYANOBACTERIAL BLOOMS ALONG THE
FRESHWATER-MARINE CONTINUUM IN A WORLD ExPERIENCING HUMAN
AND CLIMATICALLY-MEDIATED CHANGE
Hans W. Paerl1, Timothy G. Otten1, Hai Xu2, Boqiang Qin2, Guangwei Zhu2, Nathan S.Hall1
University of North Carolina at Chapel Hill, Institute of Marine Sciences, Morehead City,
NC 28557, USA
2
State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography
and Limnology, Chinese Academy of Sciences, Nanjing 210008, PR China
1
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ORAL PRESENTATIONS
Coastal watersheds support nearly 75% of the world’s human population and are
experiencing unprecedented urban, agricultural and industrial expansion. The
freshwater-marine continua draining these watersheds are increasingly experiencing
nutrient over-enrichment, eutrophication, and proliferation of harmful cyanobacterial
blooms (CyanoHABs); with negative impacts on higher plant and animal habitats as well
as animal and human health. In addressing nutrient input reductions needed to stem
and reverse this troubling trend, phosphorus (P) has traditionally received priority in
upstream freshwater regions, because some CyanoHABs can fix atmospheric nitrogen
(N2) to satisfy their nitrogen (N) requirements. Conversely, controlling nitrogen (N) inputs
has been the focus of management strategies in estuarine and coastal waters. However,
freshwater, brackish and full-salinity components of the continuum are structurally and
functionally connected. Furthermore, eutrophying freshwater and marine systems are
increasingly plagued with non N2 fixing CyanoHABs that are N and P co-limited or even
N limited. In some systems N loads are increasing faster than P loads. Therefore N and
P input constraints are likely needed for long-term CyanoHAB control. Climatic changes,
specifically warming, increased vertical stratification, salinization, and intensification
of storms and droughts, favor CyanoHABs and thus play synergistic roles in promoting
CyanoHAB frequency, intensity, geographic distribution and duration. In particular, rising
temperatures cause shifts in critical nutrient thresholds at which cyanobacterial blooms
can develop. In practical terms, this means that nutrient input reductions aimed at
controlling CyanoHABs may need to be more aggressively pursued in a warming world.
Additional control steps that should be considered include 1) altering the hydrology
to enhance vertical mixing and/or flushing and 2) decreasing nutrient fluxes from
organic rich sediments by physically oxygenating or removing the sediments or capping
sediments with clay. These efforts however have met with mixed results and can disrupt
benthic and planktonic habitats. In most instances, long-term effective eutrophication
and CyanoHAB control must consider both N and P loading dynamics within the context
of altered thermal and hydrologic regimes associated with climate change.
USING GENETIC MARKERS TO DETERMINE THE EFFECT OF SEEDING
ON THE DISTRIBUTION OF A SAxITOxIN-RESISTANT MUTATION IN
myA AREnARIA
Jennifer Phillips1 and Laurie Connell1
1
School of Marine Sciences, University of Maine, Orono, ME 04469
ORAL PRESENTATIONS
We have identified a sodium (Na+) channel mutation in softshell clams Mya arenaria
that causes a 1,000-fold decrease in binding affinity at the saxitoxin-binding site in the
sodium channel pore, thereby conferring a resistance to paralytic shellfish toxins (PSTs)
produced by the genus Alexandrium. This mutation results in improved fitness (growth,
motility and survival advantage of resistant individuals) during toxic blooms, as well as
higher toxin accumulation rates. Therefore, PSTs can act as a strong natural selection
agent in nature, leading to spread of toxin resistance in M. arenaria populations. My
work has shown that selection for this resistant allele is correlated with the prevalence
and intensity of Alexandrium spp. blooms, resulting in higher numbers of resistant
individuals in areas with regular HAB events, however the effect of juvenile Mya arenaria
seeding on the distribution of resistant and sensitive clams is currently unknown. This
study examines the effect of the seeding program at the Downeast Institute in Beals, ME
on the genetic composition of seeded sites in the northeastern US with respect to the
resistant mutation, using newly characterized M. arenaria microsatellite markers.
Page 68
SOME ASSEMBLY REQUIRED: MULTIDOMAIN PKS/NRPS GENES IN
DINOFLAGELLATES
Tsvetan R. Bachvaroff1, Ernest Williams1, Charles F. Delwiche2, and Allen R. Place1
Institute of Marine and Environmental Technology, 701 E. Pratt St., Baltimore MD 21202
Department of Cell Biology and Molecular Genetics and the Maryland Agricultural
Experiment Station, University of Maryland, College Park, MD 20742
1
2
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October 2013
ORAL PRESENTATIONS
Dinoflagellates are important marine primary producers, infamous for producing toxins,
but also significant producers of the beneficial omega three polyunsaturated fatty acids.
We undertook Illumina RNA sequencing of two species known to produce PKS-derived
toxins: Amphidinium carterae, and Karlodinium veneficum, and a third non-toxic species
Akashiwo sanguineum. In these species the toxin and fatty acid biosynthetic genes are
likely PKS derived and previous results found only single PKS domain genes that was
thought to be the norm for dinoflagellates. Surprisingly, numerous multidomain PKS,
NRPS, and hybrid PKS / NRPS genes ranging from 8 - 15 kb were found from all three
species. Six different multidomain genes were found: one contained only PKS domains,
three were hybrid NRPS /PKS genes, and two were NRPS genes. Introns were found in
PCR products amplified from genomic DNA templates in A. carterae for all but the NRPS
genes. Similarly, all categories but the NRPS had at least one species with a partial
characteristic dinoflagellate spliced leader sequence. Overall sequence assembly or
coverage for these large transcripts was poor, requiring iterative assembly techniques
in several cases. Phylogeny of KS, adenylation, AT, and KR domains from the complete
dataset suggested the multidomain genes were acquired as such, rather than derived
from single domain genes. However, the majority of sequences with PKS or NRPS
domains were encoded as single domain genes and appeared to be quite diverse in
these species. It appears that a combination of cis-multidomain proteins interacting
with trans-single domain proteins can provide the diversity of polyketides observed in
these remarkable biosynthetic organisms.
INHIBITION OF THE DINOFLAGELLATE CELL CYCLE AFTER INOCULATION
WITH THE ALGICIDAL COMPOUND IRI-160AA ExCRETED BY
SHEWAnELLA SP. IRI-160
Kaytee L. Pokrzywinski1, Mark E. Warner1 and Kathryn J. Coyne1
University of Delaware College of Earth, Ocean, and Environment, 700
Pilottown Rd., 19958 Lewes, DE, USA
ORAL PRESENTATIONS
1
Previous work in our laboratory investigated the algicidal activity of a bacterium,
Shewanella sp. strain IRI-160. We confirmed that algicidal activity was due to a secreted
compound (designated IRI-160AA) that targets dinoflagellates while having little to
no effect on other phytoplankton species. We also demonstrated that the algicide
induces programmed cell death in dinoflagellates, marked by changes in morphology,
extracellular H2O2 production, caspase-like enzyme activity, and loss of asymmetry in
the cellular membrane. Cell death induced by algicide IRI-160AA may be a consequence
of disrupting the cell cycle. The dinoflagellate genome is characterized by massive
amounts of genomic data that contain tandem repeats, no or low histone-like proteins,
no nucleosome structures, and a high abundance of transition metals and base
modifications (hydroxymethyluracil). In addition, dinoflagellates are unusual in that
they retain a permanent nuclear envelope during the cell cycle. The chromosomes also
remain condensed throughout the cell cycle and extra-nuclear microtubule spindles
traverse through cytoplasmic channels during mitosis. These features of dinoflagellates
may be a target for the algicide. Here, we used microscopy and flow cytometric analysis
to investigate the effects of the algicide on cell cycle progression in three dinoflagellates:
Prorocentrum minimum, Karlodinium veneficum and Gyrodinium instriatum. The
effect of the algicide on the cell cycle of the coccolithophore Emiliania huxleyi was
also included as a control. Our results suggest a change in cell cycle progression for
dinoflagellate cultures treated with the algicide along with nuclear degeneration and
massive degradation of chromosomal DNA, supporting the hypothesis that the algicide is
targeting dinoflagellate-specific nuclear structures and/or checkpoints that play a role in
controlling cell cycle progression of these algae.
Page 70
CIGUATOxIN AND HAWAIIAN MONK SEALS: MONITORING BLOOD
TOxIN LEVELS IN NORTHWESTERN HAWAIIAN ISLANDS SEALS AND
EVALUATION OF FORAGING UNDER STRESS USING A MOUSE MODEL
John Ramsdell1, Jessica Tiedeken1, Marie-Yasmine Bottein1, Zhihong Wang1, Aurelie
Ledreux1, Jennifer Fuquay1, Liz Kashinsky2, Angie Kaufman2, Michelle Barbieri2 and
Charles Littan2
Marine Biotoxins Program, National Centers for Coastal Ocean Science, NOAA, 219 Fort
Johnson Rd., Charleston, SC 29412, USA
2
Hawaiian Monk Seal Research Program, Pacific Islands Fisheries Science Center, NOAA,
1601 Kapiolani Blvd., Honolulu, HI 96814, USA
1
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October 2013
ORAL PRESENTATIONS
Ciguatoxin, an algal neurotoxin endemic to circumtropical regions, is believed to
be present in the food web of the Hawaiian monk seal. A critically endangered
species, the Hawaiian monk seal population is declining in its primary habitat of the
Northwestern Hawaiian Islands, and the role that ciguatoxins may play in this decline
is being investigated through several means. Several potential ciguatoxin congeners
were detected in monk seal liver samples by LC-tandem-mass spectrometry. P-CTX3C was confirmed by matching peak area ratios of 6 MRM transitions of the suspected
protonated P-CTX-3C ion (m/z 1023.6 ® 125.1, 155.1, 447.5, 907.6, 987.6, and 1005.6)
with those of P-CTX-3C standard. Ciguatoxin activity (ranging from 0.4 to 5.5 pg/ml)
was identified by Neuro2A cytotoxicity assay in whole blood of 19% of 55 free-ranging
seals collected between 2007 and 2010. Analysis of archived blood spot cards from an
expanded set of animals collected between 2003 and 2011 is ongoing to identify age
classes at highest risk of exposure. The decline of the Hawaiian monk seal is evident in
low survival of juveniles and may result from poor foraging compounded by the stress
of top level predators. We therefore developed a murine model to examine the effect
of ciguatoxin exposure and foraging under stress. A bright open field with food in the
center was used to imitate a foraging environment and restraint-induced stress a proxy
for top level predator stress. C57BL/6 mice accustomed to restraint stress over 3 weeks
were exposed to ciguatoxin P-CTX-1B (0.26 ug/kg, ip) or saline. Commencing 4 days
after ciguatoxin exposure and repeated ten days thereafter, response and behaviors in
the open field were recorded, tracked and observed, noting the time until the animal
took the food. Animals 15 and 25 days after ciguatoxin exposure that received the
restraint stress immediately prior to testing showed significantly longer time to take the
food than matched controls for either restraint stress or ciguatoxin exposure. Taken
together, these results indicate that ciguatoxin exposure is common in the Hawaiian
monk seal and can induce a disease state in mice that negatively impacts foraging in a
stressful environment.
LINKING FISH TO HUMAN ILLNESS: CONFIRMATION OF CIGUATERA FISH
POISONING CASES FROM ST.THOMAS, U.S. VIRGIN ISLANDS 2010-2012
Alison Robertson1, Lynn M. Grattan2, Sparkle Roberts2, Justin D. Liefer1, Ana C. Garcia1,
Harold A. Flores Quintana1, Jennifer I. Hooe-Rollman1, Margaret Abbott2, Elizabeth G.
Radke3, J. Glenn Morris3
Gulf Coast Seafood Laboratory, Division of Seafood Science and Technology, U.S. Food &
Drug Administration, Dauphin Island, AL 36528, USA
2
School of Medicine, University of Maryland, College Park, MD 20740, USA
3
Emerging Pathogens Institute, University of Florida, Gainesville, FL 32610, USA
ORAL PRESENTATIONS
1
Ciguatera fish poisoning (CFP) occurs throughout the tropics, particularly in island
communities such as the U.S. Virgin Islands where fish are a primary source of protein.
The impact of CFP in these communities is difficult to assess due to underreporting of
illnesses, misdiagnosis, and lack of clinical tests to confirm ciguatoxin exposure. In
St. Thomas, CFP annual incidences have been estimated at 120 per 10,000 residents.
While some symptoms of CFP are characteristic, many gastrointestinal and neurological
disturbances are quite similar to other toxin poisoning syndromes. Confirmation of CFP
cases relies on the availability of a meal remnant or leftover fish portion from which
ciguatoxins (CTX) can be confirmed using chemical methods. Retrospective toxin data such
as this was used to develop the current FDA safety guidance levels of 0.1 µg CaribbeanCTX-1 (C-CTX-1) equivalents/kg fish tissue. This study focused on CFP cases where meal
remnant toxicity data could be directly linked to human illness. Persons reporting to the
Roy Schneider Hospital, St. Thomas, between April 2010 and April 2012 with a medical
diagnosis of acute CFP were asked to participate in this study. Participants completed
a detailed questionnaire designed to provide demographic data and information on the
implicated fish including: source, species, and meal preparation. In addition, data on the
symptoms, severity, and duration of illness experienced by the participants were collected.
Of the 57 total participants during this period, 20 were able to provide a meal remnant
or leftover portion of fish that was eaten prior to the onset of acute symptoms. Meal
remnant samples were subjected to chemical extraction and examined for the presence
of ciguatera-related toxins using the sodium channel-specific mouse neuroblastoma
cytotoxicity assay and liquid chromatography tandem mass spectrometry (LC-MS/MS).
Of the 20 meal remnants tested, 16 were confirmed to contain C-CTXs ranging from 0.32.4 µg C-CTX-1 equivalents/kg fish. Multivariate analyses were used to determine factors
(e.g., demography, fish species, fish toxicity), which correlated with the overall patterns
of symptom diversity, severity, and duration reported. Snapper and barracuda were most
frequently implicated in CFP illness, with highest toxicity reported in a barracuda meal
remnant at more than 20 times the FDA guidance level (2.4 µg C-CTX-1 equivalents/kg).
Other fish implicated in case illnesses included grouper, king mackerel, horse-eye jack,
and triggerfish, which have been previously associated with CFP from this region. These
data and associated correlations will be discussed in relation to regulatory analyses and
management of CFP.
Page 72
PLANKTONIC LAYER PHENOMENA IN THE ECOLOGY OF TOxIGENIC
PSEudO-nITZSCHIA: ExAMPLES FROM COASTAL CALIFORNIA
J. P. Ryan1, J. B. J. Harvey1, Y. Zhang1, H. A. Bowers1, C. M. Mikulski2, G. J. Doucette2, C. A.
Scholin1
1
Monterey Bay Aquarium Research Institute, Moss Landing, CA, 95039, USA
NOAA/National Ocean Service, 219 Fort Johnson Rd., Charleston, SC, 29412, USA
2
Figure 1. Variations in Pseudo-nitzschia spp. abundance and particulate domoic acid (DA)
in Monterey Bay, CA, across a fontal zone where phytoplankton layer phenomena were
examined. The AUV section represents the upper 30 m over an area ~150 km2.
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October 2013
ORAL PRESENTATIONS
Observations along the eastern North Pacific show that the ecology of toxigenic Pseudonitzschia spp. is strongly influenced by phenomena of subsurface layers. Such layers
may be responsible for some “cryptic” HAB events, in which the presence of a toxic
bloom is first signaled by the appearance of poisoned animals rather than by detection
of the causative toxic bloom. Interdisciplinary study of layer phenomena is challenging,
particularly when layer vertical scales are fine. Layer ecology research is essential not
only to understanding bloom dynamics, but also to developing more effective HAB
monitoring, prediction, and management strategies. Using examples from Monterey Bay
and San Pedro Bay, California, we describe layer ecology of toxigenic Pseudo-nitzschia
spp. Results are derived from experiments conducted between 2000 and 2013. Earlier
experiments integrated ship-based sampling of phytoplankton with multidisciplinary
synoptic observations from autonomous underwater vehicles (AUVs). More recent
experiments have more closely coupled environmental and biological observations,
using autonomous environmental and molecular analytical time-series at fixed locations,
ship-based sampling, and autonomous feature recognition and targeted sampling
onboard AUVs, followed by application of molecular methods to samples. The enhanced
resolution of both environmental and biological variation enabled by these methods is
contributing to advancing this challenging research. These interdisciplinary observations
indicate processes influencing the formation and persistence of phytoplankton layer
structures, as well as population diversity, transport, nutrient inputs, and toxicity.
OPTIONS IN MITIGATING CYANOBACTERIA BLOOMS
Kevin G. Sellner1, Allen R. Place2, Michael Paolisso3, Yonghui Gao4, Ernest Williams2, Elizabeth VanDolah3.
Chesapeake Research Consortium, 645 Contees Wharf Road, Edgewater, MD 21037, USA
IMET-UMCES, 701 E. Pratt St., Baltimore, MD 21202, USA
3
Department of Anthropology, University of Maryland, College park, MD 20742, USA
4
HPL-UMCES, 2020 Horns Point Rd., Cambridge, MD 21613, USA
1
2
ORAL PRESENTATIONS
Toxic and non-toxic cyanobacteria blooms are recurrent and increasing problems in many fresh and tidal-fresh systems due to excessive nutrient loads and increasing water temperatures. Local use and exposures threaten citizens, domestic animals, and wildlife, leading to high interest in preventing or mitigating blooms. Research has been conducted for the past several years to adapt several techniques for eventual routine application in bloom mitigation, focusing on the Chesapeake region. Techniques explored in the current study include modifications of a sediment-flocculant technique successfully employed in China (Pan et al. 2006), flushing, barley straw exposure, peroxide additions, and Phoslock® application. Laboratory successes with sediment-flocculant removals of Microcystis aeruginosa (Certner et al. 2011) have been difficult to reproduce using sediment and flocculant concentrations reported from China. The last year of the project will focus on documenting the most practical and living resource protective use of sediment-flocculant additions as a routine mitigation technology. Post-bloom flushing and early spring deployment of barley straw (Hordeum vulgare) appear quite promising in limiting bloom development and toxin (microcystis and anatoxin) concentrations. The addition of peroxide shows an initial rapid decline in phytoplankton and cyanobacteria, but rapid recovery of the bloom assemblage. The addition of P-binding Phoslock® has some success when DIP levels are high but less dramatically when the nutrient pool is low and the effect is short-lived. A very interesting aspect of this work is the assessment of citizen perceptions of blooms, toxins, causes, and willingness to mitigate. In one location, high awareness of the bloom and its threat across many sectors led to effective communication with and understanding of the mitigation approaches. In another system, however, poor understanding and communication on HABs across sectors led to discord and rejection of local decisions on watershed development strategies. Implications of these communication networks, or their absence, allow recommendations of future approaches to insure public awareness and confidence in possible bloom mitigation strategies.
Certner, R. et al. 2011. Using sediment flocculation to reduce the impacts of Chesapeake Bay Microcystis aeruginosa harmful algal blooms. Thesis, GEMSTONE Program, University of Maryland, College Park, MD. pp. 194.
Pan, G. et al. 2006. Removal of harmful cyanobacterial blooms in Taihu Lake using
local soils. III. Factors affecting the removal efficiency and an in situ field experiment using chitosan-modified local soils. Environ. Pollut. 141: 206-212.
Page 74
IMPLEMENTATION OF SOLID PHASE ADSORPTION TOxIN TRACKING
(SPATT) FOR THE MONITORING HARMFUL ALGAL BLOOMS IN SOUTHERN
CALIFORNIA
Erica L. Seubert1, Alyssa G. Gellene1, Paige Connell1, Jayme Smith1, George Robertson2,
Astrid Schnetzer3 and David A. Caron1
University of Southern California, 3616 Trousdale Parkway, Los Angeles, CA, 90089, USA
Orange County Sanitation District, 10844 Ellis Ave, Fountain Valley, CA, 92708, USA
3
North Carolina State University, 2211 Hillsborough St., Raleigh, NC, 27697, USA
1
2
Page 75
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October 2013
ORAL PRESENTATIONS
The complex spatiotemporal distribution of harmful algal blooms (HABs) has
complicated our understanding of their ecology. HAB monitoring programs have existed
in many areas, but the discrete sampling may not occur on the appropriate time scale
of bloom development or in the precise location. Technologies that can increase our
spatial and temporal monitoring of HABs without requiring a significant increase in
financial commitment are of great interest. Solid phase adsorption toxin tracking
(SPATT) has been implemented in several locations worldwide for the monitoring
of dissolved phycotoxins. The SPATT resin integrates dissolved phycotoxins present
in the water over the entire deployment period, providing information on toxin
presence in an area between discrete samplings. SPATT deployments in the southern
California area began in 2012 at two coastal locations, one in the highly urbanized Los
Angeles area and the second on the sparsely developed Catalina Island, as well as the
deployment of several temporary offshore moorings in the fall of 2012 and spring of
2013. The SPATT deployment in Los Angeles harbor was in conjunction with a HAB
monitoring program in which discrete samples were collected on a weekly basis for
chlorophyll a concentrations, particulate DA and STX, dissolved inorganic nutrients,
and phytoplankton community composition. DA and STX detected in the SPATT from
this location occurred prior to and during periods in which the known producers of DA
and STX were present in the discrete samples. Information collected from the Catalina
deployment revealed relatively low to below detection concentrations of DA and STX
throughout the study period. The highest DA concentration measured in the SPATT
bag deployments was from a temporary offshore mooring, at 1 meter depth, deployed
from September 9 to September 17, 2012. The concentration of 9,200 ng DA/g resin
was markedly higher than the concentration measured in the bag deployed at 7 meters
depth on the same mooring, 39 ng DA/g resin. The results from the year and a half of
coastal SPATT deployments and the two seasons of offshore SPATT deployments will be
presented along with discussion on how the information collected has improved HAB
understanding in the region.
COORDINATING FLORIDA RED TIDE COMMUNICATION: A CASE STUDY
Dianne Shipley1, Liz Bumpus 1,Tom Higginbotham1, Hector Mendez1, Jennifer Clemente1,
Quintin Clark1, and Chuck Henry1
ORAL PRESENTATIONS
1
Florida Department of Health in Sarasota County, Sarasota, FL
Sarasota County experiences almost annual blooms of the toxic dinoflagellate, Karenia
brevis. As part of the Florida Department of Health, our mission is to protect, promote
and improve the health of all people in Florida through integrated state, county and
community efforts. Therefore, we continually work toward building connections through
partnerships with other agencies and organizations to enhance the health and safety
of our community. During the fall of 2012, we instituted a weekly conference call to
discuss the current bloom with community partners. Our goal was to provide accurate
and updated bloom status reports, facilitate consistent messaging across agencies,
monitor media articles for accuracy, and assess any outdoor recreational events (i.e.
triathlons, swim meets, etc.) where people could potentially be at high-risk for exposure
to the bloom. We invited partners from Mote Marine Laboratory, FWC/Fish and Wildlife
Research Institute, NOAA HAB Bulletin, USF Center for Red Tide Prediction, Florida
Department of Health Aquatic Toxins Program, Florida Department of Aquaculture and
Consumer Services, Sarasota County Government Communications, Sarasota County
Environmental Utilities, four area chambers of commerce, and Sarasota Convention and
Visitor’s Bureau (Visit Sarasota County) to participate on the conference calls. As the
bloom expanded, we invited partners from other impacted counties to join as well. This
is the first time an effort for consistent health messaging on Florida red tide has been
conducted across five counties (Manatee, Sarasota, Collier, Lee and Monroe). We were
able to utilize the expertise of many different people which provided us with a significant
pool of knowledge that helped in making our message as reliable and well-informed
as possible. We believe the calls decreased reactionary media reporting, and improved
resident and visitor knowledge about how to stay healthy during a Florida red tide. The
calls concluded in March 2013 as the bloom also ended.
Page 76
EVALUATION OF POTENTIAL OYSTER CONTAMINATION FROM
CYANOBACTERIAL TOxINS
Emily A. Smith, Chase Weidert, Sibel Bargu
Department of Oceanography and Coastal Sciences, School of the Coast and
Environment, Louisiana State University, Baton Rouge, LA, 70803
Page 77
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October 2013
ORAL PRESENTATIONS
The occurrence of harmful algal blooms in estuaries has increased around the world
with the potential for their toxins to enter the food web of these ecosystems. One type
of vector through which algal toxins can enter the estuarine food web is filter-feeding
organisms such as mussels and oysters. Northern Gulf of Mexico, especially Louisiana
(LA) has a very large oyster market. The oysters are harvested from the estuaries that
consist of fresh water in the upper areas and have an increasing salinity gradient as
they approach the Gulf of Mexico. Oysters are usually found in the 10-20ppt area of
the estuary. Freshwater cyanobacteria move into the outer more saline waters when
diversions from the Mississippi River are opened in the spring. The question then arises
as to whether the toxin producing cyanobacteria can survive in higher saline waters
and whether the oysters can uptake their toxins. One of the most commonly seen
toxin producing cyanobacteria, Anabaena, was grown at increasing salinities (0-10ppt)
for 10 days. Anabaena grew well at 7ppt or lower salinity. It did not grow, but did not
decline in biomass at 8ppt and above indicating the ability to maintain biomass even
in higher saline waters. Particulate (intracellular) and dissolved (extracellular) toxins
were measured at the beginning and end of the experiment. Particulate toxins initially
were below the detection limit but increased over time in all salinities except 9 and
10ppt. Dissolved toxins were also below the detection limit initially, but were then
detected at the end of the experiment with the highest concentration in 10ppt (0.74
± 0.49 microcystins µg/L) treatment. Oysters collected from Breton Sound Estuary, LA,
were then allowed to feed on toxin producing Anabaena over a three-hour period. The
experimental oysters’ initial consumption rate was 3.5x105 ± 2.0x105 cells consumed
oyster-1 minute-1 but decreased over time. Analysis of the oyster viscera at the beginning
and end of the experiment showed an increase amount of toxins in the oysters. The
results indicate the ability of Anabaena to not only grow and produce toxins at higher
salinities, but also that oysters can feed on these toxin producing cyanobacteria and
contain their toxin. Since toxin producing cyanobacteria can be natural members of the
phytoplankton community in estuaries, threat of contamination due to their toxins is
imminent and monitoring should be a priority for these ecosystems.
A SYSTEMS BIOLOGY APPROACH TO UNDERSTANDING mICROCySTIS
BLOOMS
Morgan M. Steffen1, T. Chad Effler1, Loren J. Hauser1,2, Zhou Li1, Rachel M. Adams1,
Brian D. Dill2,3, Nathan C. VerBerkmoes2,4, Stephen P. Dearth1, Amanda L. May1,
Shawn R. Campagna1, B. Shafer Belisle1, Gregory L. Boyer5, Sue B Watson6, Richard A
Bourbonniere6, Steven W. Wilhelm1
University of Tennessee, Knoxville, TN 37996, USA
Oak Ridge National Lab, Oak Ridge, TN 37830 USA
3
University of Dundee, Dundee DD1 5EH, United Kingdom
4
New England BioLabs, Ipswich, MA 01938 USA
5
State University of New York, Syracuse NY 13210 USA
6
Environment Canada, Burlington, ON L7R 4A6 Canada
1
ORAL PRESENTATIONS
2
Recurrent blooms of the toxic cyanobacterium Microcystis have plagued Lake Erie for
decades. Recent expansion in size and duration of toxic bloom events has galvanized
the effort to identify the factors that drive the success of Microcystis and other toxic
cyanobacteria in the environment. To obtain a better understanding of the molecular
response of Microcystis to different environmental conditions, we have taken a
comprehensive systems biology approach. The suite of tools employed include
metagenomics, metatranscriptomics, metaproteomics, and metametabolomics,
applying each to cultured isolates and field samples collected from Lake Erie. Analyses
have focused on the interactions between Microcystis and available nutrients, but
include genomic information on how the co-occurring microbial community responds
to, and perhaps shapes, environmental conditions. Initial findings resulting from
metaproteomics implicated upregulation of the Microcystis urease enzyme in the
environment. Recent advances in metatranscriptomics and metametabolomics indicate
a complex cellular response when the compound urea is available to Microcystis as a
nitrogen source. To obtain these data, triplicate samples from sites in Lake Erie were
sequenced on the Illumina® HiSeq platform. Sequences were then analyzed to compare
expression differences of Microcystis across multiple environmental gradients, as well
as to simulated variable nutrient conditions tested on culture isolates. Differences in
composition and expression patterns of associated heterotrophic bacteria were also
considered, as these are thought to influence the response of Microcystis to a variable
environment. Comprehensive surveys of total small metabolites were run in tandem
with metatranscriptomes to generate a secondary measure of physiological response
to diverse environmental conditions. Taken together, these observations provide novel
and extensive insight into the complex cellular and community interactions that take
place during cyanobacterial blooms and may highlight previously unknown molecular
mechanisms driving Microcystis bloom events.
Page 78
PERCEPTIONS OF BEACH ACTIVITIES DURING FLORIDA RED TIDE
BLOOMS: A CONJOINT ANALYSIS
Jamie L Studts1, Margaret M Byrne2, Kate Kohler3, Barbara Kirkpatrick2,3,4
Dept of Behavioral Science, Univ. of Kentucky College of Medicine, Lexington, KY
Dept of Public Health Sciences, Miller School of Medicine, Univ. of Miami, Miami, FL
3
Mote Marine Laboratory, Sarasota, FL
4
Rosenstiel School of Marine and Atmospheric Sciences, Univ. of Miami, Miami, FL
1
2
Page 79
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October 2013
ORAL PRESENTATIONS
Background: Communicating accurate information regarding harmful algal blooms
(HABs) presents a significant public health challenge. This is certainly true with Florida
red tide, i.e., Karenia brevis, blooms. In recent research supported by the National
Institute of Environmental Health Sciences (NIEHS) and others, the investigative team
found that Florida red tide caused both acute and possibly chronic health effects,
as well as significant socioeconomic impacts to coastal residents and visitors. Pilot
work revealed that although approximately 70-80% of residents and tourists reported
knowledge of Florida red tide, fewer than 50% had correct information about the
potential health risks of Florida red tide for asthmatics.
Methods: We conducted semi-structured interviews to establish the key concerns
about beach activities during a Florida red tide. Subsequently, we developed a conjoint
survey and administered it to 303 local and seasonal residents (i.e., snowbirds) using
Sawtooth Software. The sample was also stratified by pulmonary health status (healthy
vs. asthmatic). Surveys were conducted from June 2011 through May 2012. The beach
activities portion of the conjoint valuation included 25 scenarios, depicting six key
attributes of decisions to go to the beach during a bloom: amount of dead fish on the
beach, posted beach warnings, odor, parking availability, and wind direction.
Results: The relative propensity to go to the beach during a bloom was 3.18 (±1.11) on
a Likert scale of 1 (low) to 9 (high). The top utilities measuring the attribute desirability
were the presence or absence of odor (81.99), the absence of dead fish (80.92), the
presence of many (versus few) dead fish (63.72), and the lack of a beach warning sign
(61.10). The relative importance scores for going to the beach during a bloom were
highest for the amount of dead fish on the beach and odor. Combining these two
attributes explained 64% of the variance in consumer decision making. When examining
associations between group status, demographic characteristics, health status, and
decision making utilities, a number of interesting patterns emerged.
Discussion: Various systems are in place to warn of the impact of Florida red tide on
beach conditions. Future studies are needed to examine the utility of these systems
and explore novel methods to disseminate accurate risk information to the beach going
community. Data from this study suggest the important role of beach clean-up efforts
that may be very relevant to the tourism industry.
Conclusion: Under-protective and overprotective reactions to a Florida red tide bloom
can adversely impact health and economic considerations, respectively. It is important
to understand factors that influence individuals’ decisions during a Florida red tide, so
that accurate risk information can be communicated and steps can be taken to mitigate
barriers to beach-going under appropriate conditions. Educational interventions
targeting community stakeholders responsible for beach maintenance and clean up may
be particularly important.
CLIMATOLOGICAL ANALYSIS OF BLOOMS IN LAKE ERIE AND SEASONAL
FORECAST
Richard P. Stumpf1, Timothy T. Wynne1, Michelle C. Tomlinson1, R. Peter Richards2, David
Baker2.
NOAA National Centers for Coastal Ocean Science, Silver Spring, MD 20910, USA
Heidelberg University, National Center for Water Quality Research, Tiffin OH 44883,
USA
1
ORAL PRESENTATIONS
2
Starting in 2003, Lake Erie’s Western Basin saw a recurrence of annual cyanobacterial
blooms. The summers of 2008 to 2011, in particular, had severe blooms. In contrast,
2012 had a respite with a milder bloom. Using data from the MERIS and MODIS sensors,
we have characterized the severity and patterns of the blooms through each summer
from 2002 to 2012 in order to understand the blooms. The analysis allows estimates
of the area and the biomass of the blooms. Combining this data with discharge and
nutrient data from the Maumee River (the major tributary to the Western Basin) has
allowed development of a model to predict bloom severity.
The inter-annual variation in bloom biomass or severity is described by variations in
spring (March to June) discharge and phosphorus loads from the Maumee River, with
the major bloom years of 2008-2011 having the highest spring flows. 2011 had both the
most severe bloom and the most extreme spring river loadings. The seasonal model has
an uncertainty of ~15% of the observed bloom intensity in years having strong blooms.
The model was was used to forecast the 2012 bloom. The forecast correctly identified
a milder bloom than 2011, however, the bloom was stronger than estimated by the
model. 2012 appears to have been an unusual year in several regards. The Maumee
River had the lowest spring river discharge since 1975 and one of the lowest total
phosphorus loads of the past 15 years. The 2012 season followed the most severe
bloom in decades (2011 event), occurred following a winter in which the lake did not
freeze and, and also followed fall and winter seasons having extremely high river flows
and loadings. This combination of events has not been previously observed. Mild
blooms have occurred during years with no ice (2002 and 2006), and during the summer
following a wet winter (2007). A forecast was planned to be made in July for the 2013
bloom severity. As the winter of 2013 had typical loadings from the Maumee and the
lake had ice cover, 2013 will provide insight into stability and robustness of the model for
predicting bloom severity.
Page 80
GAmbIERdISCuS BIOGEOGRAPHY AND THE CONSEQUENCES OF
CLIMATE CHANGE
Patricia A. Tester1, Steven R. Kibler1, William C. Holland1 and Mark W. Vandersea1, Emma
L. Hickerson2 and R. Wayne Litaker1
1
Center for Coastal Fisheries and Habitat Research, National Centers for Coastal Ocean
Science, National Ocean Service, NOAA, 101 Pivers Island, Beaufort, NC, 28516, USA.
2
Office of National Marine Sanctuaries, Flower Garden Banks National Marine
Sanctuary, NOAA. 4700 Avenue U, Galveston, Texas, 77551, USA
Gambierdiscus is the dinoflagellate genus identified most closely with the production of
ciguatoxins and the outbreaks of ciguatera fish poisoning (CFP). While CFP has not had
the attention or visibility of other HAB-related illnesses, it claims more victims than all
other HAB-caused maladies combined. Following the taxonomic revision of the genus
and the development of species-specific qPCR assays there has been a resurgence
of interest in Gambierdiscus research. It is now possible to examine Gambierdiscus
species diversity in tropical areas and expand our ideas of suitable habitat to include the
Mediterranean Sea, Canary Islands and the main islands of Japan. Currently, G. australes,
G. pacificus, G. polynesiensis, G. toxicus and G. yasumotoi appear to be endemic to
the Pacific while G. excentricus, G. ruetzleri and Gambierdiscus ribotypes 1 and 2 are
known from the Caribbean Sea, Atlantic Ocean and Mediterranean Sea. Gambierdiscus
belizeanus, G. caribaeus, G. carolinianus and G. carpenteri have been found in both the
Atlantic and Pacific Oceans.
http://gambierdiscuswiki.wikispaces.com/
Page 81
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October 2013
ORAL PRESENTATIONS
In regional scale studies it is not unusual to see a suite of Gambierdiscus species in small
areas or with overlapping distributions of toxic and non-toxic species with different
temperature preferences. For example, the Flower Garden Banks National Marine
Sanctuary (27°56′N, 93°36′W) in the northern Gulf of Mexico has an area of only150
km2 but this habitat supports six of the seven Gambierdiscus species (and ribotypes)
found in the Caribbean. While there is species-specific variation, Gambierdiscus
growth rates typically increase significantly as temperatures rise from 25 to 31o C.
Projections from a variety of climate models indicate the northern Gulf of Mexico will
experience an increase of 2.1-2.2 °C to depths of 200 m by 2100. A one degree increase
in water temperature is projected to add ~26 days of optimal growth conditions for
Gambierdiscus species per year in the northern Gulf of Mexico seaward of the 100 meter
isobath. The more substantial projected increase of ~2 °C will add an average of 51-55
days of optimal growth conditions and reduce or eliminate temperatures cold enough to
cause Gambierdiscus mortality. It is important to note that even the slight temperature
increases projected for the northern Gulf of Mexico within the next few decades portend
greater CFP risk.
GLOBAL ANALYSIS OF MRNA HALF-LIVES AND dE nOvO
TRANSCRIPTION IN THE FLORIDA RED TIDE DINOFLAGELLATE, KAREnIA
bREvIS
Frances M. Van Dolah and Jeanine S. Morey
ORAL PRESENTATIONS
NOAA Marine Biotoxins Program, Center for Coastal Environmental Health and
Biomolecular Research, Charleston, SC 29412
The regulation of gene expression underlies all cellular processes resulting in the growth,
toxicity, and demise of harmful algal blooms. In most eukaryotes, transcription is a
principal point of gene regulation. However, dinoflagellates possess many processes
that appear to be under post-transcriptional control, and number of characteristics
that suggest that gene expression is regulated predominantly at the translational
level. However, the extent to which their genes are regulated post-transcriptionally
remains unresolved. To gain insight into the relative roles of differential mRNA stability
and de novo transcription in dinoflagellates, we biosynthetically labeled RNA with
4-thiouracil to isolate newly transcribed and pre-existing RNA pools in Karenia brevis.
The isolated fractions were then used for analysis of global mRNA stability and de novo
transcription by hybridization to a K. brevis microarray. Global K. brevis mRNA half-lives
were calculated from the ratio of newly transcribed to pre-existing RNA using the online
software HALO (Half-life Organizer). Overall, mRNA half-lives were substantially longer
than reported in other organisms studied at the global level, ranging from 42 minutes to
greater than 144 h, with a median of 33 hours. Consistent with well-documented trends
observed in other organisms, housekeeping processes, including energy metabolism
and transport, were significantly enriched among the most highly stable messages.
Shorter-lived transcripts included a higher proportion of genes involved in RNA
regulation, stress response, and other response/regulatory processes. One such family
of proteins involved in post-transcriptional regulation in chloroplasts and mitochondria,
the pentatricopeptide repeat (PPR) proteins, had dramatically shorter half-lives when
compared to the arrayed transcriptome. We have previously observed that PPR protein
transcripts are among the most rapidly increasing transcripts in response to nutrient
addition. We therefore queried the newly synthesized RNA pools at 1 and 4 h following
nitrate addition to N-depleted cultures to determine if these changes were the result
of de novo transcription or increased RNA stability. Transcriptome-wide there was little
evidence of changes in the rates of de novo transcription during the first 4 h following
N addition, and no evidence for increased PPR protein transcription. This suggests that
their rapid increase in abundance following N addition results from increased message
stability, not new transcription. These results lend support to the growing consensus of
post-transcriptional control of gene expression in dinoflagellates.
Page 82
ExAMINING THE RESPONSE OF HARMFUL DINOFLAGELLATES TO
THE BACTERIAL ALGICIDE IRI-160AA: DIFFERENCES IN GROWTH,
PHYSIOLOGY AND MODE OF ACTION ACROSS LABORATORY AND FIELD
COLLECTED SAMPLES
Mark E. Warner, Charles L. Tilney, Kaytee L. Pokrzywinksi, Kathryn J. Coyne
University of Delaware, College of Earth, Ocean, and Environment, 700 Pilottown Rd.,
Lewes, DE 19958, USA
Page 83
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October 2013
ORAL PRESENTATIONS
As harmful algal blooms continue to be a persistent problem in many coastal areas,
effective algicidal agents are one tool that may prove useful in controlling some algal
species. However, establishing the mode of action and specificity of response for
particular phytoplankton as well as efficacy in natural mixed algal communities remains
challenging. We have begun to characterize the algicidal activity of several small
compounds found in the filtrate of the bacterium Shewanella sp. IRI-160AA that have a
high specificity for inhibiting growth of at least 12 dinoflagellates, including Gyrodinium
instriatum, Karlodinium veneficum, Prorocentrum minimum, and Alexandrium
tamarense. While there is a strong relationship between the presence or degree of
thecal armoring and the level of algicidal activity, the temporal nature of inhibition
and pathways of cellular damage appear different across these species. Growth
arrest as well photoinactivation followed a dose dependent response in G. instriatum
and K. veneficum. Proxies for programed cell death, photoinactivation, and loss of
membrane integrity appear rapidly and occur simultaneously in G. instriatum, while a
dark dependent mode of photoinactivation occurs in K. veneficum prior to the loss of
membrane integrity and cell lysis. Further, microscopic examination of K. veneficum
following higher algicide dose showed changes in chloroplast morphology, followed
by chloroplast expulsion. Short-term algicide applications were tested against freshly
collected samples of natural blooms of G. instriatum, K. veneficum and P. minimum.
Contrary to the expected results, there was a slight increase in P. minimum cell density
following application of the algicide, and growth arrest took longer in samples of G.
instriatum and K. veneficum compared to laboratory culture experiments. Further
analysis of dose response also indicated a much higher concentration of the algicide was
needed as compared to laboratory tests. Molecular analyses of the plankton community
showed an increase in raphidophyte and diatom biomass over time that subsequently
obscured our ability to accurately detect any algicide dependent photochemical changes
to the target dinoflagellates. In addition to changes in phytoplankton number and type,
several experiments resulted in an increase in the number of bactivorous ciliates (e.g.
Paraphysomonas spp. and Euplotes sp.) that may have resulted from increased bacterial
abundance following the release of DOM by dinoflagellates. While the initial tests with
natural bloom samples were not as clear as those with isolated laboratory cultures,
there was still promising evidence for specific dinoflagellate algicidal activity that will be
examined at a larger scale in in situ mesocosm experiments.
SPEED TALK
PRESENTATIONS
SPEED ORAL
TALK PRESENTATIONS
S
NOAA’S ECOLOGICAL FORECASTING ROADMAP
Allison L. Allen1, Paul A. Sandifer2, and Richard P. Stumpf1
National Oceanic and Atmospheric Administration, 1305 East-West Highway, Silver Spring
MD 20910, USA
2
National Oceanic and Atmospheric Administration, 331 Fort Johnson Road, Charleston
SC 29412, USA
1
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
For more than a decade, a number of NOAA offices have been researching ecological
processes and developing experimental forecasts for a variety of ecosystem
components, including harmful algal blooms, pathogens, jellyfish, brown shrimp,
hypoxia, distributions of habitat and key species, sea level change, wave energy, and
ocean acidification (oceanservice.noaa.gov/observations/ecoforecast). The most mature
of these are harmful algal bloom (HAB) forecasts, which are fully operational in the Gulf
of Mexico with the HAB Operational Forecast System (HAB-OFS) .
Historically, however, NOAA has lacked a formal Ecological Forecasting Program, which
raises challenges in identifying corporate priorities, securing long-term resource
commitments and maximizing the efficiency of existing forecasting abilities. In 2012,
NOAA developed the framework for an Ecological Forecasting Roadmap a prioritized,
agency-wide approach for coordinating NOAA’s existing capabilities and operational
environmental prediction and service delivery infrastructures. Such a broadly-supported
NOAA strategy for ecological forecasting will offer management solutions to assure
protection, maintenance and restoration of the health and productivity of ocean, coastal
and Great Lakes ecosystems, for both natural resources and human communities. This
agency-wide effort will also help connect internal-NOAA activities to related efforts in
the external academic community and private sector.
Within the Roadmap, there are three initial NOAA priorities, HABs, hypoxia and
pathogens. These were chosen based on the following: their economic, environmental
and health effects, importance to constituents and to the environment, and apparent
increasing prevalence, distribution and intensity in numerous areas of the country. The
Roadmap will address the key issues relevant to HAB forecasts, including: sustained
resources, improved models to improve forecast accuracy and resolution, additional
data collection from both remote and in situ sensors, upgraded computing capacity
to process these data and enhanced forecast dissemination using NOAA-wide
communications assets.
The outcome of the Roadmap will be a suite of more accurate and reliable forecasts in
all regions of the country concerned with harmful algal blooms, hypoxia, and pathogens.
As the Roadmap effort progresses, customers should see more dependable, higher
quality products on a broader scale than currently available under the current ad hoc
development and delivery model.
Page 86
ASSESSING PRE-BLOOM, SUB-SURFACE POPULATIONS OF PSEudOnITZSCHIA IN THE SAN PEDRO SHELF REGION OF SOUTHERN
CALIFORNIA
Holly A. Bowers1, Roman Marin III1, John P. Ryan1, G. Jason Smith2, Raphael Kudela3,
Christina M. Mikulski4, Gregory J. Doucette4 and Christopher A. Scholin1.
Monterey Bay Aquarium Research Institute, 7700 Sandholdt Road, Moss Landing, CA
95039, USA
2
Moss Landing Marine Laboratories, 8272 Moss Landing Road, Moss Landing, CA 95039, USA
3
Ocean Sciences Department, University of California Santa Cruz, 1156 High Street, Santa
Cruz, CA 95064, USA
4
NOAA/National Ocean Service, P.O. Box 12607219 Ft. Johnson Road, Charleston, SC
29412, USA
1
In order to better understand the diversity and spatial distribution of these sub-surface
populations on the SPS using molecular methods and microscopy, water samples
were collected via shipboard net tows, CTD casts, and autonomously targeted AUV
deployments during spring 2013. During the mission, samples were collected using an
adaptive sampling strategy that permitted us to explore diversity throughout features
of interest (e.g., high chlorophyll patches) across shelf waters and the adjacent deep
channel. The experiment fortuitously spanned a pre-bloom period in which numerous
Pseudo-nitzschia spp. comprised a fraction (less than twenty percent) of the diatomdominated phytoplankton assemblage. This non-bloom scenario allowed us to establish
clonal cultures from a wide variety of Pseudo-nitzschia species and to interrogate those
for DA production. These findings were compared to samples collected in the area a few
weeks later during a bloom characterized by high Pseudo-nitzschia cell concentrations
and DA.
October 2013
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In situ detection of Pseudo-nitzschia and DA during these types of missions is carried
out on moored Environmental Sampling Processors (ESPs). The probe-based array for
Pseudo-nitzschia detection on the ESP targets two common DA producers: P. australis
and P. multiseries. During several ESP deployments, including the most recent one at
the SPS, we have observed a discrepancy in the molecular probes used to detect P.
multiseries in conjunction with spikes in DA, indicating the possible presence of ‘cryptic’
toxigenic species. Since shipboard sampling had ended before the spring 2013 ‘event’
in San Pedro Bay, we explored recovery of frustules from spent ESP extraction filters for
TEM/SEM to identify ‘cryptic’ Pseudo-nitzschia species.
SPEED ORAL
TALK PRESENTATIONS
During the past decade, the San Pedro shelf (SPS) region in southern California has
become a hotspot for Pseudo-nitzschia blooms and domoic acid (DA) intoxication of
marine mammals and birds. Interestingly, harmful impacts may “appear” suddenly, with
no indication of increasing cell abundances or DA from routinely monitored shore-based
stations. In part, this discrepancy is due to widespread spatial and temporal distribution
of blooms in that region. However, other factors are hypothesized to be involved,
including advection of sub-surface populations. The presence of Pseudo-nitzschia
populations in sub-surface layers has been described along the greater NE Pacific. Layer
ecology for these species in Monterey Bay and the SPS region is an active research area
(see Ryan et al. and Smith et al., this conference).
PERCEPTIONS OF SEAFOOD CONSUMPTION DURING FLORIDA RED TIDE
BLOOMS: A CONJOINT ANALYSIS
Margaret M Byrne1, Jamie L Studts2, Kate Kohler3, and Barbara Kirkpatrick1,3,4
Dept of Public Health Services, Miller School of Medicine, Univ. of Miami, Miami, FL
Department of Behavioral Science, University of Kentucky College of Medicine,
Lexington, KY
3
4
Rosenstiel School of Marine and Atmospheric Sciences, Univ. of Miami, Miami, FL
1
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
2
Background: Misperceptions about the causes and effects of harmful algal blooms
(HABs) abound. These misperceptions are especially of concern when they affect
individuals’ actions in such a way that social or economic harms are incurred. Our pilot
work found that only about 50% of tourists and residents had correct information about
the safety of eating seafood during a Florida red tide, i.e., Karenia brevis.
Methods: Semi-structured interviews established the key concerns citizens have
regarding eating seafood during a Florida red tide. A survey including a conjoint
valuation analysis (CVA) was developed and conducted using Sawtooth Software among
303 local residents and seasonal residents (i.e., snowbirds), stratified by lung health.
Participants had to eat seafood to participate in the study. Surveys were conducted from
June 2011 through May 2012. The attributes of the 24 seafood CVA scenarios were:
government warnings on seafood safety, trust in the retailer, price, local versus chain
retailer, and origin of the seafood (Gulf of Mexico or not).
Results: General propensity to eat seafood during a bloom was 4.27 (stdev 1.14) on
a Likert scale of 1 (low) to 9 (high). The most important attributes from the CVA in
decisions about seafood were a government warning explaining 41% of the variation
in decisions, and trust in the merchant, 24%. Price was 3rd in importance, explaining
18% of variance. Logistic regressions with importance scores as the dependent variable
were conducted to assess whether demographics were associated with importance
of attributes. We found that purchasing locally was significantly less important for
asthmatics than those with healthy lungs (p<0.047), and that trust in the retailer was
significantly more important for younger respondents (p<0.026). General propensity to
eat seafood during a Florida red tide was significantly lower for women (p<0.001).
Discussion: Florida Division of Aquaculture and Florida Fish and Wildlife Research
Institute are tasked with monitoring and closing of shellfish beds during Florida red
tide blooms. We found that warnings or reassurances from the government were
by far the most important factor in individuals’ decisions regarding eating seafood.
However, overall propensity to eat seafood was relatively low. Better messaging, possibly
targeted for some demographic populations, on the safety of consuming seafood that
is commercially harvested is important to avoid deleterious economic effects during a
Florida red tide.
Conclusion: It is important to understand what affects individuals’ decisions during a
red tide, so that appropriate education and outreach can be developed. Inappropriate
reactions to a Florida red tide event can have negative economic consequences.
Page 88
CELLULAR RESPONSE TO OSMOTIC STRESS IN KAREnIA bREvIS
Lisa Campbell1, Reagan Errera1, Darcie Ryan1, and Andrea Bourdelais2
Texas A&M University, 3146 TAMU Dept. Oceanography, College Station, TX 77843
University of North Carolina, Center for Marine Science, Wilmington, NC 28409
1
2
Osmotic stress initiates a rapid response by an organism to minimize cellular damage,
followed by a longer-term response to re-equilibrate the cell with its new environment.
Previous studies have demonstrated that in response to a rapid decrease in salinity,
Karenia brevis, the major harmful dinoflagellate species in the Gulf of Mexico, produced
up to 53% more brevetoxin. While the role of brevetoxin in this response is not known,
the correlation between brevetoxin production and salinity suggests that osmotic
sensing is occurring. Ion transport would be an integral component of the osmotic
response to the stress.
Transcriptome results also included one predicted monogalactosyldiacylglycerol (MGDG)
synthase. MGDG is the most abundant lipid in thylakoid membranes. The observed
2-fold change in comparison with control indicated upregulation of the MGDG synthase
1 hr after hypoosmotic stress. Initial metabolome results for the hypoosmotic stress
experiments with K. brevis reported a product with similar retention time and atomic
mass as MGDG that was not seen in the control. Since glycolipids such as MGDGs are
found in the cell and chloroplast membranes, it is possible that the K. brevis cells can
also change the composition of their cell membranes rapidly in response to salinity
changes. Additional results from metabolome analyses will be discussed. Focus on
changes in the metabolome of stressed treatments vs. controls will complement the
transcriptome results by providing validation of signaling/metabolic pathways.
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SPEED ORAL
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An interdisciplinary approach combining physiological experiments, imaging-in-flow
cytometry, next generation sequencing, and metabolomic profiling was employed to
examine the responses of three clones of K. brevis to osmotic changes. Transcriptome
analysis identified two types of transmembrane proteins that facilitate osmoacclimation
in all clones: aquaporins and voltage-gated ion channels. Aquaporins are a family of
major intrinsic proteins that are sensitive to osmotic pressure and provide a mechanism
for rapid transport of water across a membrane to adjust cell volume. A number of
voltage-gated ion channels (K+, Na+, Ca2+) were also identified. Because voltage-gated
ion channels are sensitive to ion gradients, they would also be involved in osmolytic
adjustment. To examine the potential for a specific ion (K+, Cl-, Na1+, Ca2+) to trigger the
osmotic response, we systematically diluted one ion while keeping the remaining ions
at equivalent molar concentration for salinity of 35. Dilution of K+ triggered a significant
increase in the production of brevetoxins, while dilution of Cl- and Ca2+ did not trigger
a significant change. These observations suggest that K. brevis may be responding to
changes in membrane potential established by the dilution of K+ during osmotic stress.
BREVETOxIN PHOTOAFFINITY PROBES FOR THE IDENTIFICATION OF
THE NATIVE BREVETOxIN RECEPTOR
Ryan Cassell and Kathleen S. Rein
Department of Chemistry and Biochemistry,
Florida International University, Miami, FL 33199
SPEEDPRESENTATIONS
TALK PRESENTATIONS
ORAL
The dinoflagellate Karenia brevis produces brevetoxin, a potent neurotoxin responsible
for substantial amounts of marine mammal and fish mortalities. When the toxin is
consumed in humans, it causes Neurotoxic Shellfish Poisoning (NSP). The native function
of brevetoxin has remained mysterious since its discovery. It has been reported that
an increase in toxin production is triggered from changes in salinity and osmotic stress
implicating a role of osmoregulation within the organism. Photoaffinity labeling has
become increasingly popular in identifying ligand receptors. By attaching ligands to these
photophors, one is able to activate the molecule (generate a carbene by exposure to UV
light) after the ligand binds to its receptor to obtain a permanent linkage between the
two. Subsequent purification will provide the protein with the ligand directly attached.
The synthesis of a brevetoxin photoaffinity probes and their application to identify a
native brevetoxin receptor will be described.
Page 90
A SATELLITE-DERIVED PREDICTIVE MODEL OF RED TIDE SEVERITY FOR
THE WEST FLORIDA SHELF
Mary C. Christman1, John Walter2, Jan Landsberg3, Brian Linton2, Karen Steidinger3,
Richard Stumpf4 and Jacob Tustison3
MCC Statistical Consulting LLC. 2219 NW 23rd Ter. Gainesville, FL 32605
Southeast Fisheries Science Center. 75 Virginia Beach Drive, Miami, FL 33149
3
100 Eighth Ave SE, St. Petersburg, FL 33701
4
NOAA, National Ocean Service, 1305 East-West Highway, Room 9115, Silver Spring, MD,
20910
1
2
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A vexing problem for HAB research is to determine the relative intensity of blooms from
one year to another. Given the event-response nature of most monitoring, cell count
data usually does not unbiasedly index bloom intensity nor does it have comprehensive
spatial and temporal coverage. In this paper we model the probability of a bloom (cell
counts >10E5 cells/liter) as a function of a suite of satellite-derived remote sensing
products and other covariates using a generalized additive model (GAM). The model is
derived using matched SeaWIFS satellite data products and FWRI harmful algal bloom
(HAB) samples for 1998-2010. The GAM approach incorporates non-linear relationships
and interactions between model factors. Predictions from the model using daily satellite
imagery provides spatial and temporally explicit mapping of the probability of a bloom
and associated prediction error over a grid covering the West Florida Shelf. When
restricted to offshore waters (>10 meters), model performance was quite strong with
44% of the total deviance explained with an overall misclassification rate of 17.4%.
In inshore waters the model performance degraded as satellite observations were
increasingly confounded by littoral dynamics as evidenced by higher prediction errors
and poorer overall performance of the model (37% explained deviance and 21% overall
misclassification rate) when estimated for the entire region. Averaging spatial and
temporal predictions over several different spatial domains (inshore, offshore of Tampa
Bay, entire region) provided annual indices of red tide intensity. Over the 12 year time
period, the 2005 probability of a bloom averaged over all locations was 37% higher than
the next highest year (2003) confirming the severity of the 2005 red tide event and
quantitatively distinguishing it from other years.
STRATIFICATION INFLUENCES ON THE MIGRATORY BEHAVIOR OF THE
HARMFUL ALGA KAREnIA bREvIS
Matthew J. Garrett1, Gary L. Hitchcock2, Kendra L. Daly3, Mark E. Luther3
Florida Fish & Wildlife Conservation Commission, 100 8th Avenue SE, St. Petersburg, FL
33701, USA
2
University of Miami, Rosenstiel School of Marine and Atmospheric Science, 4600
Rickenbacker Cswy, Miami, FL 33149, USA
3
University of South Florida, 830 1st St. SE, St. Petersburg, FL 33701, USA
SPEEDPRESENTATIONS
TALK PRESENTATIONS
ORAL
1
Migratory behavior has been documented in many phytoplankton species, including the
harmful dinoflagellate Karenia brevis. Diel vertical migration may offer a competitive
advantage to K. brevis population by increasing exposure to favorable nutrient and light
regimes. However, the degree to which the strength of stratification in the physical
environment influences migratory behavior of K. brevis is not well understood. In this
study, we examined the effects of a weak and strong stratification on the migratory
behavior of K brevis through the introduction of a thermocline in a controlled
experimental water column. Experiments were conducted experiments in a 2m PVC
cylinder in a temperature (25°C) and light (110 μE/cm-1sec-1) controlled incubator.
Migratory behavior was documented under three conditions: 1) homogenous and
unstratified, 2) weak thermally stratified, and 3) strong thermally stratified. Vertical
stratification was established by wrapping tubing around the lower half of the column
and circulating water through a thermal chiller such that the temperature of the lower
column was controlled to create a two temperature and density layered system The
column was filled with nutrient replete water, inoculated with a culture of K. brevis,
and the experiment was run for 48 hours. To measure the vertical position of K.
brevis, we used four continuously recording Turner Cyclops 7 fluorometers affixed at
different depths. Stratification altered the migratory behavior of K. brevis. This work
will contribute to an understanding the alteration of migratory behavior of naturally
occurring blooms in the context of predictive modeling.
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REAL-TIME WATER QUALITY MONITORING FOR HAB EVENTS USING
AUTONOMOUS PORTABLE WATER LABORATORIES AND DATA
VISUALIZATION “CLOUD” ANALYSIS
Brian Gregson1, Katherine Hubbard2, Paul Janecek1 and David Fries1
1
Spyglass Technologies, Inc., 101 16th Ave South, Ste 4A, St. Petersburg, FL 33701 USA
100 8th Avenue SE, St. Petersburg, FL 33701 USA
2
A number of unique automated water sampling and analytical platforms incorporate
biological and chemical sensors to broaden utility for water quality monitoring applications,
and are currently in use in harmful algal bloom (HAB) monitoring applications. Presented
here is a review of novel monitoring instruments as well as ancillary equipment and data
analytical services to support water quality monitoring activities, especially related to
HABs.
The Porifera Automated Field Sampler allows for solid-phase concentration, archival and
retrieval of chemical, biological and particulate samples for in-situ fluorescence and/or
shored-based interdisciplinary analyses.
Underwater mass spectrometers (UMS), both ion-trap and quadrupole, employ proprietary
novel sample inlets for high-fidelity real-time in-situ monitoring of volatile (and limited
semi-volatile) chemical targets of interest. Results are shown demonstrating utility of
UMS for HAB monitoring applications.
To enable actionable responses based on data provided by such a diverse “fleet” of
instrumentation, the Spyglass Water Information Portal provides visualization of data
feeds from the described equipment, or as a standalone service for 3rd-party sensors.
The interactive and user-friendly cloud-based software service provides researchers,
regulatory managers and other stakeholders powerful and rapid decision-making tools.
This flexibility in instrumentation and support services facilitates routine detection of
HAB-relevant compounds as well as relevant water quality parameters—in situ and in near
real-time—thus providing novel insight into the dynamics and ecology of HAB species.
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The Spyglass Environmental Sample Processor (ESP) has been deployed in open ocean and
coastal environments since 2001. Analytical methods available for the ESP are designed
to target taxa of interest, and include molecular probe arrays, immunoassays (cELISA),
and quantitative Polymerase Chain Reaction (qPCR). Reagent manufacturing services are
available in accordance with Good Laboratory Practices Guidelines, as well as support
equipment for lab-based development and ground-truthing of new and/or improved
assays.
REAL-TIME DETECTION OF HARMFUL ALGAE AT A TRIBAL MARINE
AQUACULTURE SITE
Marco B.A. Hatch1, Stephanie K. Moore2, Linda Rhodes2, William Nilsson2, Chris Scholin3,
Steven Hallam4, Annie Cox4, Karl Mueller5, Vera Trainer2 and Mark Strom2
Salish Sea Research Center, Northwest Indian College, 2522 Kwina Road, Bellingham,
WA 98226 2Northwest Fisheries Science Center, 2725 Montlake Blvd E, Seattle, WA
98112 3Monterey Bay Aquarium Research Institute, 7700 Sandholdt Rd, Moss Landing,
CA 95039 4University of British Columbia, Department of Microbiology and Immunology,
Life Sciences Center, 2350 Health Sciences Mall, Vancouver, BC V6T-1Z3 5Lummi Natural
Resources, 2616 Kwina Road, Bellingham, WA 98226
1
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
Cultured marine shellfish and finfish are vulnerable to naturally occurring harmful algal
blooms (HABs) that produce toxins. The economic cost of toxic HABs to coastal tribal
communities may be more difficult to evaluate; however, given the importance of
seafood to tribal culture and economies, it is likely to be significant. In Puget Sound,
concern over increasing HAB frequency and the emergence of new HAB threats calls for
improvements in sample collection and analysis.
The Environmental Sample Processor (ESP) is an autonomous sampling and analysis unit
that employs DNA-based technology to detect microorganisms in water samples. It is
positioned on site, and relays data by telemetry. The ESP uses sensitive and specific
molecular assays to produce quantitative results. In 2012, researchers deployed an ESP
at Friday Harbor, north of Puget Sound, for the very first time and targeted one of the
primary fish-killing HAB species, Heterosigma akashiwo. H. akashiwo was detected in
the water column at low levels during the entire 44-day deployment from June through
July. Twice during the deployment, H. akahsiwo abundance approached levels known to
kill fish. The ESP provided early warning of these events and the information was rapidly
disseminated to stakeholders and triggered increased site surveillance at fish farms
throughout Puget Sound and British Columbia.
In 2013, two ESP units will be deployed in tandem in Lummi Bay, in northern Puget
Sound, for a prolonged real-time assessment of Heterosigma, Alexandrium spp., Pseudonitzschia spp. These units will be in position from June through August, sampling
daily. Marine waters and shellfish will be sampled weekly for toxin levels. The primary
objective is to determine if the ESP can provide early warning of outbreaks of pathogens
that impact shellfish and finfish aquaculture at a tribal shellfish and finfish hatchery in
an area with extensive subsistence harvesting and a history of the targeted HABs. A
secondary objective is to determine if relationships exist between HAB levels in marine
waters and toxin levels in shellfish tissues. The ESP enables extensive high frequency
sampling that is necessary to determine such relationships and to detect increases in
populations of targeted pathogens in marine waters before they are able to contaminate
or harm shellfish or finfish. This collaborative effort brings researchers from NOAA, UBC,
MBARI, Northwest Indian College, and Lummi Natural Resources together to address
issues that may limit exercising of inherent rights. Two American Indian students from
Northwest Indian College will also act independent researchers on this project.
Page 94
CHANS: ENVIRONMENTAL FACTORS INFLUENCING KAREnIA bREvIS
BLOOMS ON THE FLORIDA WEST COAST: INTERPRETATION IN SUPPORT
OF HEALTH AND SOCIO-ECONOMIC IMPACTS
Gary Hitchcock2, Vincent J. Lovko1, Gary Kirkpatrick1, Barbara Kirkpatrick1, Kate Kohler1,
Zoe Shoesmith2, Amy Clement2.
Mote Marine Laboratory, 1600 Ken Thompson Drive, Sarasota, FL
RSMAS, 4600 Rickenbacker Causeway, University of Miami, Miami, FL
1
2
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As a component of the NSF Coupled Natural and Human Systems program, our focus
is to elucidate linkages between harmful algal blooms and associated impacts on
human health, social, and economic aspects of the coastal communities between
Tampa and Ft Myers, FL. Identifying these linkages is challenging, since it also requires
an understanding of the potential human influence on coastal physical, chemical, and
biological processes that impact bloom dynamics. One major objective is to determine
if relationships exist that are potentially mediated through management actions. A
major goal of this objective is to determine if relationships exist between precipitation,
watershed discharge and bloom dynamics. An initial study examined twelve watersheds
along the Florida west coast from 2002 to 2008. During that interval there were
two years each with low, average, and above-average precipitation. In general, the
magnitude of annual bloom maximum cell abundance exhibited a weak, but positive,
relationship with total coastal precipitation and watershed discharge. However, there
was a very limited correspondence between the cell abundance and distribution with
precipitation, discharge at monthly time scales. A second goal is to determine if specific
environmental conditions are related to indices of respiratory irritation. A change from
no detectable to mild respiratory irritation at several west Florida beaches was found to
correspond to air temperature, water temperature, and relative humidity. These factors
suggest that the environmental factors that regulate aerosolization influence the transfer
of brevetoxin from the ocean’s surface to the coastal atmosphere. We also assisted the
project goals through the interpretation and sorting of cell counts via the FWC/Florida
Wildlife Research Institute to assure the cell counts used in the analysis are pertinent
to the project, since the database is a compilation of all sampled cell counts. As an
example, cell counts were sorted from the East coast of Florida in the bloom of 2007 to
compare the bloom movement from north Jacksonville) to south with the Agency for
Health Care Administration (AHCA) inpatient admissions for the same time period.
Multidisciplinary projects such as this one challenge the team to discuss and analyze
data sets together to assure accurate data analysis and interpretation.
STRESS RESPONSE IN KAREnIA bREvIS: CHANGES IN RIBOSOMAL RNA
David S. Jayroe1 and Timothy I. McLean1
University of Southern Mississippi, Department of Biological Sciences, 118 College Drive
#5018, Hattiesburg, MS, 39406-0001, USA
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
1
Karenia brevis is a marine dinoflagellate that causes harmful algal blooms in the
Gulf of Mexico. These blooms are responsible for massive fish kills, shellfish bed
contaminations, adverse human health effects, and vast economic loss. In order to
understand higher order bloom behavior and dynamics it is imperative to understand
K. brevis at the cellular level. In growing K. brevis in the laboratory under a variety of
conditions, we have noted a distinct shift in the size of both ribosomal RNAs upon
culturing cells under “stress” conditions, namely depleted nutrients, cold shock, and
decreased salinity. Interestingly the large ribosomal subunit becomes larger in size,
and the small ribosomal subunit becomes smaller. This response by K. brevis had not
been previously described in the literature. These RNAs, pre- and post-stress, are being
fully sequenced to determine how they are different, and what mechanisms may be
responsible for producing them: alternative splicing? different transcriptional initiation
or termination sites? different loci? These results will help us understand the molecular
events surrounding K. brevis survival under certain environmental conditions, which
may have implications regarding K. brevis biogeographical distribution and bloom
termination.
Page 96
PHOTOCATALYTIC DEGRADATION OF mICROCySTIn-LR BY ROSE
BENGAL
Wenjun Jiang1, Dionysios D. Dionysiou2, Kevin E. O’Shea1
Department of Chemistry and Biochemistry, Florida International University, Miami, FL,
33199, USA
2
Environmental Engineering and Science Program, University of Cincinnati, Cincinnati, OH
45221-0012, USA
1
The bloom-forming cyanobacteria can produce and release cyanobacterial toxin in
freshwater bodies, and microcystin (cyclic heptapeptides) is the most common toxin.
Microcystins are reported to be potential tumor promoters. While approximately 80
different variants of microcystins have been isolated and identified, microcystin-LR (MCLR) is one of the most toxic and problematic variants.
Under photolysis by visible or solar light no measureable degradation of MC-LR was
observed in the absence of RB, however the concentration of MC-LR is significantly reduced
under such conditions in the presence of RB. The degradation of MC-LR in presence of 500
µM RB follows the pseudo-first order kinetic model nicely. These results indicate RB can
be used as a photosensitizer (catalyst) to effectively degrade MC-LR. Rate of degradation
increase three fold under oxygen saturated conditions compared to argon-saturated
conditions, demonstrating that photo-oxidation is the predominant degradation process.
Under argon saturation the degradation is likely due to the energy transfer between the
RB* and MC-LR. The results of detailed kinetic studies using furfuryl alcohol and ongoing
product studies will be presented. The results can also help understand the photochemical
transformation of MC-LR by dissolved organic matter in natural environments.
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Conventional water treatment methods are often not effective or not practical for the
removal of MC-LR. In this study, we employed rose bengal (RB), a singlet oxygen (1O2)
sensitizer, to study the photo-oxidative destruction of MC-LR. 1O2 is reactive oxygen
species, which can react with unsaturated organic compounds via ene-type reaction,
[2+2] cycloaddition, and Diels-Alder reaction.
FROM SOCIAL MEDIA TO NATIONAL WEATHER SERVICE PRODUCTS:
ExPLORING NEW OUTREACH TOOLS FOR NOAA’S HARMFUL ALGAL
BLOOM OPERATIONAL FORECAST SYSTEM
Karen Kavanaugh1, Edward Davis1, Katherine Derner2, and Adria Schneck-Scott1
NOAA, National Ocean Service, Center for Operational Oceanographic Products and
Services (CO-OPS), Silver Spring, MD 20910, USA
2
NOAA, National Ocean Service, CO-OPS, 672 Independence Parkway, Chesapeake, VA
23320, USA
SPEEDPRESENTATIONS
TALK PRESENTATIONS
ORAL
1
Since it was transitioned from research to operations in 2004, NOAA’s Harmful Algal
Bloom Operational Forecast System (HAB-OFS) has provided year-round operational
forecasts of the potential for Karenia brevis bloom formation, transport, intensification
and associated respiratory irritation for the Gulf of Mexico region. Although the HABOFS has focused on maintaining high quality forecasts and refining the forecast system
through research and development, the continued success of the HAB-OFS is also
dependent on reaching its target audiences with useful information. The forecasts
are communicated through two main products, the public conditions report and the
technical HAB bulletin. These products are both intended as decision support tools: one
for the general public and the other for a more technical audience that includes coastal
resource managers, public health officials and researchers. After almost ten years in
operations, it remains a challenge to sustain communication with current audiences,
while attracting new interest.
Beginning in August 2012, the HAB-OFS team launched a coordinated outreach strategy
with the goal of increasing awareness of the HAB-OFS program and its products among
potential HAB bulletin users and the general public. First, the HAB-OFS Quarterly
Newsletter was introduced as a way of providing updates on activities and new product
developments, answering frequently asked questions, and giving additional product
background. Next, a Facebook page was created as another means to disseminate public
conditions reports, network with HAB-OFS partners and inspire interest in HABs from
non-scientific individuals, especially those who live in or visit the Gulf of Mexico. Most
recently, beginning in February 2013, in cooperation with NOAA’s National Weather
Service (NWS), HAB forecasts for “high” levels of respiratory irritation have joined
other hazards covered by the NWS Beach Hazards Statement test product, allowing the
HAB-OFS to reach an even broader audience with important public health information
through a variety of means including the NWS home page and NOAA Weather Radio.
Preliminary results of these outreach efforts already show a greater than 15% increase
in HAB bulletin subscriptions, approximately 1.5x the number of visits to the HAB-OFS
website, and a consistent monthly increase in the number of people interacting on the
Facebook page. In order to develop a sustainable and successful outreach strategy,
the HAB-OFS has and will continue to rely on collaboration, coordination and feedback
from our partners on location in Florida and Texas. Through increased communication
with our partners and the public, these outreach efforts have already resulted in
important refinements to the original HAB-OFS products and identified goals for future
enhancements.
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THE APPLICATION OF IMMUNO-HISTOCHEMISTRY AS A TOOL FOR
INVESTIGATING SUSPECTED, HISTORIC, OR UNRECOGNIZED HABRELATED ANIMAL MORTALITY AND DISEASE EVENTS
Jan H. Landsberg1, Theresa Cody1, and Patrick Wilson1
1
Immuno-histochemistry (IHC) is a useful histological technique that is used extensively in
medical and veterinary diagnostics to visualize the presence of specific targeted antigens
in tissues. IHC can identify the distribution and organotropism of toxins in animals
and can aid in the assessment and interpretation of toxin-associated histopathological
effects. IHC is also useful to supplement or support confirmatory analytical methods
such as LC-MS or HPLC. For example, IHC can visually demonstrate the presence of
toxins, such as microcystins, that are covalently bound in tissues, and which are normally
difficult to extract for analysis by rapid or confirmatory assays such as ELISA or LC-MS.
In some cases, IHC may have application in the detection of low levels of toxins where
traditional analyses are constrained by minimal levels of detection.
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We have successfully used IHC to visualize the presence of microcystins and brevetoxins
in a range of animal species (fish, birds, and marine mammals) and tissues during and
after HAB events. This technique has application for investigating unexplained animal
mortality or disease events where HABs were suspected or unknown, but toxins could
not be confirmed or results were equivocal in tissues. Further, IHC can also be used
for routine animal mortality investigations as a screening tool for differential diagnosis
and for toxin rule outs. IHC can be used to retrospectively detect toxins (if stable) in
archived histological tissues from historical cases with unsolved or suspected HAB
etiologies. IHC is also useful for detecting subtle histopathological changes associated
with chronic or low level exposures to phycotoxins and to discern their sublethal effects
on animals. While IHC has its advantages, it should not be used exclusively without
other confirmatory analyses for toxin presence, and considerable methods development
and testing is required to avoid misinterpretation of false positives, especially in the
absence of field supported data. Further, there are differences in species’ toxicokinetics,
cross-reactivities, and histopathological responses to HAB toxins that can confound
the interpretation of the results, demonstrating the need for validation of the IHC with
experimental exposures.
SURVEY OF CARIBBEAN CIGUATOxINS IN FISH COMMONLY CONSUMED
IN ST. THOMAS, US VIRGIN ISLANDS
Christopher R. Loeffler1, David A. Olsen2 and Alison Robertson1
1
FDA, Gulf Coast Seafood Laboratory Dauphin Island, AL 36528, USA
St. Thomas Fishermen’s Association, St. Thomas, US Virgin Islands 00802, VI
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
2
Ciguatera fish poisoning (CFP) is endemic to the US Virgin Islands (USVI). Illness
estimates in St. Thomas range from 600-4000 people annually. In St. Thomas, the
primary CFP prevention measure is to utilize local fishermen’s knowledge and experience
in avoiding species and fishing grounds previously implicated in CFP. Unfortunately,
illnesses are still relatively common and without the benefit of reliable rapid field tests
and species-specific toxin prevalence rates, reducing the risk of CFP will be difficult. This
study employed four geolocations to evaluate the prevalence of Caribbean ciguatoxins
(C-CTXs) in four fish species that are commonly found in local markets of St. Thomas,
USVI; Balistes vetula (queen triggerfish), Ocyurus chrysurus (yellowtail snapper),
Epinephelus guttatus (red hind), and Haemulon plumierii (white grunt). These species
were targeted for collection by fishermen using standard fishing practices, at historic
and commonly utilized locations. C-CTXs were extracted from 100 g sub-samples of
homogenized fish muscle. Extracts were analyzed by neuro-2a neuroblastoma cell assay,
and confirmed using liquid chromatography tandem mass spectrometry. High variability
in C-CTXs was observed between and within geolocations, resulting in a sporadic
distribution of fish containing detectable levels of toxins. Of the four species surveyed to
date, queen trigger and red hind had the highest prevalence of ciguatoxins (~40%). The
identification of C-CTXs in fish commonly marketed in St. Thomas highlights a potential
exposure risk to local consumers. This information will inform local fisheries resource
managers and public health agencies in the development of CFP prevention strategies.
Page 100
CHANS: DEVELOPMENT OF A HAZARD MEASUREMENT SYSTEM FOR
FLORIDA RED TIDE
Vincent J. Lovko1, Derrick S. Hudson1,2, Gary Hitchcock3, Porter Hoagland4, Barbara A.
Kirkpatrick1,3,5, Gary Kirkpatrick1, Richard P. Stumpf6
University of South Florida, Tampa, FL
3
Rosenstiel School of Marine and Atmospheric Sciences, University of Miami, Miami, FL
4
Marine Policy Center, Woods Hole Oceanographic Institution, Woods Hole, MA
5
Dept of Epidemiology and Public Health, Miller School of Medicine, University of
Miami, Miami, FL
6
NOAA National Ocean Service, Silver Spring, MD
1
2
Page 101
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SPEEDORAL
TALK PRESENTATIONS
PRESENTATIONS
The impact of natural hazards on human and natural resources can be difficult to
measure. The use of hazard scales and indices (such as those used for wind events,
tropical storm systems, and earthquakes) provide both a method of classifying the
severity of specific hazard events and a predictive capability for determining impact. The
nearly seasonal occurrence of blooms of Karenia brevis on the West Florida shelf are
typically reported as a location-specific measure of cell abundance. While this measure
can provide an indication of the severity and extent of a bloom and of the immediate
potential risk of respiratory illness, it is not sufficient to provide a comprehensive
assessment of the overall severity of a bloom or the longer-term predictive capability
of hazard risk. As a component of a comprehensive, multi-institutional coupled human
and natural systems (CHANS) research effort, we describe the initial development of a
harmful algal bloom hazard measurement system to assist in assessing and interpreting
the impacts of Florida red tide on the communities and natural resources of the central
west coast of Florida. As an initial step in this effort, we use measures of the temporal
and spatial aspects of bloom manifestation, including aerial coverage of bloom, duration
of bloom stages, bloom intensity (based on cell and toxin concentrations) and proximity
to coastal resources, to ascribe a bloom severity value to recent and past bloom events.
Using impact data derived from multiple sources (e.g. Beach Conditions Reporting
System, economic data collected as part of the overall CHANS effort) we examine the
correlation of this bloom severity value to effects on human resources. When fully
developed, this tool could serve as a public outreach product to respond to media and
community queries concerning the comparative severity of current and past bloom
events. Further development, application, and assessment of this hazard evaluation
system will be discussed.
THE vIbRIO-HAb CONNECTION: INVESTIGATING THE INFLUENCE OF
IRON ON FORMATION OF vIbRIO BIOFILMS AND ROS PRODUCTION BY
HETEROSIGmA AKASHIWO
Christopher R. Main1 and Kathryn J. Coyne1
University of Delaware College of Earth, Ocean and Environment, Lewes, DE 19958
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
1
Bacteria within the genus Vibrio are found throughout coastal and marine environments
where they often form associations, such as biofilms, with phytoplankton and other
eukaryotic organisms. Prior research in our lab demonstrated a close association
between Vibrio cholerae and the raphidophyte, Heterosigma akashiwo during blooms
of this species, and microscopic examination confirmed the attachment and formation
of biofilms by Vibrio cholerae on a local strain of H. akashiwo. Iron is required for the
formation of biofilms by Vibrio spp. and enhances the growth and persistence of Vibrio
in the natural environment. Bacterially produced siderophores, high affinity iron ligands,
also increase planktonic uptake of iron and may influence iron availability in the euphotic
zone. Photolysis of the siderophore vibrioferrin, for example, releases iron as Fe(II),
and oxidation of the Fe(II) to Fe(III) allows for uptake by both the algae and associated
bacteria. Low iron concentrations have also been shown to enhance the production of
reactive oxygen species in the harmful raphidophyte, Heterosigma akashiwo, which may
contribute to its toxicity toward fish and shellfish. Here, we investigated the role of iron
on the formation of biofilms by Vibrio cholerae and the interaction of iron and Vibrio on
production of ROS by Heterosigma akashiwo. Cultures of H. akashiwo were acclimated
to a range of Fe concentrations and mixed with non-pathogenic Vibrio cholerae.
Samples were collected prior to inoculation, and at specific intervals up to 24 hours after
inoculation for determination of ROS production. Cellular attachment of V. cholerae
was then evaluated at 24 hours after inoculation and the ratio of attached V. cholerae
to H. akashiwo was determined by qPCR. Research is ongoing, but preliminary results
indicate a negative correlation between biofilm formation of Vibrio on Heterosigma cells
and ROS production. The influence of iron and presence of Vibrio on the transcriptome
of H. akashiwo is also being investigated and results will be presented. Results of
this research will increase our understanding of potential health risks associated with
Heterosigma blooms and inform future management practices in this region.
Page 102
FLORIDA’S FIRST SHELLFISH HARVEST CLOSURE DUE TO DOMOIC ACID
Sheila N. O’Dea1, Leanne J. Flewelling1, Jennifer Wolny1, Julie Brame1, Karen Henschen1,
Paula Scott1, Katherine A. Hubbard1, Jeff Wren2, Carrie Jones2 Christopher Knight2, and
Christopher Brooks2
Florida Fish and Wildlife Conservation Commission, Fish and Wildlife Research Institute,
100 Eighth Avenue Southeast, St. Petersburg, FL 33701, US
2
Florida Department of Agriculture and Consumer Services, Division of Aquaculture
1203 Governor’s Square Blvd, Suite 501, Tallahassee, FL 32301, US
1
The Pseudo-nitzschia bloom peaked in early May and was relatively short-lived. The
maximum observed Pseudo-nitzschia abundance during the bloom was 14.7 x 106
cells/L, and total DA concentrations reached up to 31.2 µg DA/L in seawater. Electron
microscopy and genetic analyses indicated that P. cuspidata was the dominant species.
By mid-May, multiple Pseudo-nitzschia species were present and cell densities had
decreased to 3.6-8.5 x 105 cells/L. The bloom had subsided by late May, with DA either
not detected or present at only trace levels in seawater.
Throughout May and June, DA was monitored in oysters, bay scallops (Argopecten
irradians), and pen shells (Atrina rigida). During the bloom, DA concentrations exceeded
the action limit in all three shellfish species. Immediately following the bloom decline
in late May, DA concentrations in oysters and scallops decreased to below the action
limit (0.1- 5 µg DA/g). Concentrations in pen shells, however, remained elevated for
several weeks, and slowly decreased to acceptable levels by June 25. In late May, finfish
representing at least 11 species and covering a range of trophic levels were collected.
Domoic acid was detected at low levels in 93% (51/55) of fish tested (individual or
pooled samples), with average concentrations of 0.07 µg DA/g in muscle and 0.2 µg DA/g
in viscera. St. Joseph Bay was reopened for shellfish harvest in early July. This brief but
significant event marks the first ever closure of a shellfish harvesting area in the Gulf of
Mexico due to DA. Given that Pseudo-nitzschia blooms are occurring more frequently in
the northern Gulf (Parsons et al., 2002) and DA production by Pseudo-nitzschia can vary
with conditions, monitoring of shellfish for DA in this region should be enhanced.
Page 103
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October 2013
SPEED ORAL
TALK PRESENTATIONS
Domoic acid (DA), a potent neurotoxin produced by members of the marine diatom
genus Pseudo-nitzschia, has the potential to cause amnesic shellfish poisoning. High
abundances of Pseudo-nitzschia (> 106 cells/L) occur frequently throughout the year
along Florida’s Gulf Coast although concentrations of DA in seawater and biota typically
remain low. In late April 2013, high Pseudo-nitzschia cell densities (> 4.9 x 106 cells/L)
in St. Joseph Bay in the Florida Panhandle triggered shellfish collection by the Florida
Department of Agriculture and Consumer Services. Oysters (Crassostrea virginica)
collected on May 6 contained 76 µg DA/g shellfish (almost 4-fold higher than the federal
action limit of 20 µg DA/g) initiating the first closure of a shellfish harvesting area in
Florida due to DA.
PROPOSED HARMFUL ALGAL BLOOM CONTROL TECHNOLOGIES:
FLOATING DESALINATION AND WATER PUMPING PLANTS
Kevin C. Owen1, Jenna L. Owen2, and Daniel P. Owen3
Port Dolphin Energy, LLC, Höegh LNG AS, 400 North Tampa Street, Tampa, FL 33602
Tinedale-Oliver & Associates, Inc., 1000 N. Ashley Dr, Tampa, FL 33602
3
Department of Biological Sciences, Florida State University, 600 W College Ave, 32306,
USA
1
2
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
Over the last several decades, there has been a dramatic increase in the areas affected
by harmful algal blooms (HABs); the number and frequencies of the HABs that have
occurred; and the economic losses attributed to HABs. The number of toxic species
identified as forming HABs and the number of toxins identified in HAB-causing species
have also increased.
A number of mechanical, biological, chemical, genetic, and environmental HAB control
technologies have been proposed. Given the characteristics of HABs and dinoflagellate
life cycles, the primary objectives of HAB control strategies should be to disrupt the
environmental conditions that support the persistence and propagation of HABs; stop
dinoflagellate cell division and, thereby, HAB population growth; induce the formation
of temporary cysts for dinoflagellates that have this life cycle stage; and, most
drastically, cause cell mortality in the HAB-causing dinoflagellates.
The authors of this paper suggest using floating desalination plants to treat or control
HABs. In a strongly stratified water column, floating water pumping plants can be used
for control. HAB-producing dinoflagellates are sensitive to such physical and chemical
conditions as changes in temperature and salinity. The discharges from a floating
desalination and water pumping plant can rapidly change the water temperature and
salinity. These changes could be expected to induce encystment in the dinoflagellate
species that form cyst and could cause mortality in those species unable to form
temporary cysts. Preventing population growth, inducing encystment, or causing
mortality would effectively end a HAB.
Discharges from a desalination plant are temporary in nature and include hypersaline
water, freshwater (hyposaline water), and heated water. The water intake, coupled
with the hyposaline and hypersaline discharges, would create advection currents and
alter the water temperature, salinity and stratification. In a strongly stratified water
column exists, pumping water could alter the temperature and salinity and disrupt
stratification without the need for additional desalination. These sudden and significant
environmental manipulations could stress and disrupt a HAB. With continued mixing
after discharge, the water would return to ambient temperature and salinity relatively
quickly with minimal effect on the marine environment. The temperature and salinity
changes created by a floating desalination or water pumping plant would be achieved
without the discharge of chemicals or other foreign materials into the environment.
Since dinoflagellates have been shown to react quickly to environmental changes, the
temperature and salinity of the discharges could be controlled to target the HAB species
and reduce adverse impacts on other marine organisms. Bench-scale and field tests
should establish optimal temperature and salinity ranges for HAB treatment and control.
Page 104
AN INVESTIGATION INTO THE EFFECTS OF SHORELINE USE ON
CYANOBACTERIAL ABUNDANCE IN SODUS BAY, LAKE ONTARIO
Katherine A. Perri1 & Gregory L. Boyer1
State University of New York - College of Environmental Science and Forestry, 1 Forestry
Drive, Syracuse, NY 13210, USA
1
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SPEED ORAL
TALK PRESENTATIONS
Greater Sodus Bay is a large shallow embayment located on the southern shore of
Lake Ontario. Sodus Bay is also home to a large recreational boating fleet, serviced by
nine marinas located around the bay. In the last three years, the Bay has experienced
repeated outbreaks of toxic cyanobacteria including Microcystis and Anabaena species.
To evaluate the impact of shoreline use on these pelagic algal blooms, water samples
were collected weekly for nutrient, pigment and toxin analysis from four marina sites,
two residential sites and the public beach located in the Village of Sodus Point during
the summers of 2011 and 2012. The Bay experienced several cyanobacterial blooms in
both summers and differences were observed between the two years. Cyanobacterial
abundance peaked in September of both years however smaller isolated blooms were
often observed early in the season. Toxin concentrations for microcystins were higher
in 2011 than in 2012. Total microcystin concentrations ranged between 0.3 - 17 µg/L
and there was considerable variability between the different marina, residential, and
beach sites. However, correlations between the toxin concentrations, nutrients, and
environmental variables at these separate sites did not support the conclusion that
marina operations were a strong source of nutrients to drive these cyanobacterial
blooms. Therefore, other factors such as weather conditions and variable anthropogenic
activities will need to be considered.
SAxITOxIN GENE STRUCTURE AND REPRESENTATION IN NON-TOxIC
AND TOxIC ALExAndRIum TAmAREnSE SPECIES
Kathleen Pitz1, Michael Brosnahan1, and Don Anderson1
Woods Hole Oceanographic Institution, 266 Woods Hole Road, Woods Hole, MA 02543
USA.
SPEEDPRESENTATIONS
TALK PRESENTATIONS
ORAL
1
Several dinoflagellates within the species complex Alexandrium tamarense produce
saxitoxins, a group of neurotoxins that cause paralytic shellfish poisoning, arguably the
most widespread HAB poisoning syndrome worldwide. We explored saxitoxin gene
diversity over the entire phylogeny of A. tamarense as well as in Alexandrium affine
and Alexandrium ostenfeldii. We compared the sequences of assembled contigs that
had strong homology to three recently described dinoflagellate genes (sxtA, sxtB, and
sxtG) that are likely to be involved in saxitoxin biosynthesis. The contigs were collected
from meta-assemblies of fifteen A. tamarense transcriptomes representing all five A.
tamarense species: eight toxic Group I clones, one non-toxic Group II clone, three nontoxic Group III clones, two toxic Group IV clones and a non-toxic Group V clone. The
contigs were recovered from the meta-assemblies by reciprocal BLAST searches using
the recently described dinoflagellate homologs as queries. Strand-specific sequencing
was used on eleven of these transcriptomes and reads were mapped back onto
contigs in order to determine presence of antisense RNA transcripts. A certain level of
uncertainty exists in meta-assembly analysis due to errors including fragmentation and
misassembly of expressed genes. We verified presence of saxitoxin genes in genomic
DNA and further examined an A. tamarense proteome for presence of saxitoxin proteins.
By examining relationships among these groups and their complement of saxitoxin genes
we can establish differences between the toxic and non-toxic species, illustrating genetic
changes that may be important for toxin biosynthesis and further elaborating on the
complexity of the saxitoxin gene cluster in dinoflagellate transcriptomes.
Page 106
ISOLATION OF BACTERIA CAPABLE OF DEGRADING MICROCYSTIN-LR
Lauren E. Krausfeldt1, Robbie M. Martin1, Helena L. Pound1, Justine R. Schmidt2, Gary R.
LeCleir1, Gregory L. Boyer2, Steven W. Wilhelm1
Department of Microbiology, The University of Tennessee, Knoxville, TN. USA.
Department of Chemistry, State University of New York, Syracuse, NY. USA.
1
2
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TALK PRESENTATIONS
Cyanobacterial harmful algal blooms (cHABs) occur worldwide, causing detrimental
effects to aquatic ecosystems and impacting local economies. These cHABs can
produce toxins that pose threats to human health and limit potable water supplies.
Microcystin-LR (MCLR) is a potent hepatotoxin and protein phosphatase inhibitor
produced by Microcystis sp. and other genera of cyanobacteria. Conventional municipal
water treatment techniques are ineffective at removing MCLR from water sources.
Heterotrophic bacteria co-occurring with harmful algal blooms have been found to
degrade and utilize MCLR as a carbon source. Published protocols using ethanol as
carrier solvent for MCLR were used to screen for and isolate potential MCLR-degrading
bacteria. High performance liquid chromatography (HPLC) analysis revealed that
ethanol was not a suitable carrier, as it provided an additional source of carbon. Using
deionized, sterilized water as the MCLR solvent, additional screenings were conducted.
Several strains of bacteria were successfully isolated from water samples of Lake Tai,
China, which has a history of recurring harmful cyanobacterial blooms dominated by
Microcystis sp. One strain, identified as Pseudomonas alcaligenes, degraded up to 90%
of MCLR toxin over a 10-day period. Ongoing degradation assays will characterize MCLR
degradation rates and byproducts with the long-term goal of identifying a bacterial
consortium that can be deployed as part of engineered toxin remediation strategies.
CHANS: INCIDENCE OF NEUROTOxIC SHELLFISH POISONINGS DURING
FLORIDA RED TIDE (KAREnIA bREvIS) BLOOMS: IS SHELLFISH
MANAGEMENT EFFECTIVE AT MITIGATING OUTBREAKS?
Jeremy Faris1, Andrew Beet2, Rebecca Lazensky3, Andrew Reich3 and Porter Hoagland2
University of Florida; College of Veterinary Medicine; College of Public Health and
Health Professions, Gainesville, Florida, US
2
Marine Policy Center, Woods Hole Oceanographic Institution, Woods Hole,
Massachusetts, US
3
Bureau of Epidemiology, Florida Department of Health, Tallahassee, Florida, US
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
1
Neurotoxic shellfish poisoning (NSP) is a poisoning in humans and other marine animals
with a range of acute neurologic, gastrointestinal, respiratory, and cardiac symptoms.
NSP can cause significant morbidity; whether it can cause chronic illness in humans
is unknown. NSP is caused by the consumption of molluscan shellfish that contain
accumulated neurotoxins, called brevetoxins, produced by the marine dinoflagellate,
Karenia brevis. This algae causes “Florida red tides” a type of harmful algal bloom (HAB),
which commonly occurs off Florida’s southwest coast. Historically, it has been observed
that most Florida NSP cases have been associated with the consumption of hard clams
(Mercenaria mercenaria) collected from shellfish harvesting areas that were closed
during active Florida red tides.
In order to determine whether NSP cases occur when Florida shellfish harvesting
areas are closed due to K. brevis-related blooms, a dataset of NSP cases reported
from 1997-2009 was compiled using case reports found in the Florida Department
of Health’s (FDOH) electronic notifiable disease database, “Merlin.” Onset-of-illness
dates were compared to shellfish harvesting area closure periods. The number of NSP
illnesses was small (n=21), representing a conservative estimate, as only those cases
reported to FDOH and meeting the strict case definition for NSP have been included.
The implementation of shellfish bed harvest closures during red tide events as a
management tool provides significant public health benefits.
Page 108
USE OF DISSOLVED INORGANIC AND ORGANIC PHOSPHORUS BY THE
TOxIC DINOFLAGELLATES KAREnIA bREvIS AND KAREnIA mIKImOTOI
(DINOPHYCEAE)
Bill Richardson
Fish and Wildlife Research Institute, Florida Fish and Wildlife Conservation Commission
100 8th Avenue SE, Saint Petersburg, Florida 33701
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SPEED ORAL
TALK PRESENTATIONS
Large harmful algal blooms of Karenia brevis and Karenia mikimotoi are thought
to persist, in part, due to their ability to use various forms of dissolved organic and
inorganic phosphorus (DOP, DIP). These species’ ability to grow on different forms of
phosphorus (P) was evaluated in the laboratory using two K. brevis clones (CCFWC 257,
CCMP 2281) and a single K. mikimotoi clone (CCFWC 67) grown axenically and nonaxenically on 28 different P compounds. Axenic and non-axenic cultures grew similarly
well on all forms of DIP and the DOP phosphomonoester compounds, although final
yields on DIP generally exceeded those on DOP. Notably, no single compound provided
exceptionally superior growth rate or yield. Axenic and non-axenic cultures differed in
their ability to use certain compounds. The axenic clones were unable to grow on the
phosphomonoester phytate, any of the phosphodiesters, or phosphonates, whereas
the non-axenic clones grew on phytate, most of the phosphodiesters, phosphonates
2-aminoethylphosphonic acid, and phosphonoacetic acid. Furthermore, differences
between axenic and non-axenic cultures suggested that bacterial degradation of some
DOP compounds was necessary before P use by Karenia, and that bacterial assemblages
in non-axenic Karenia cultures differed in their exploitation of DOP compounds. Results
suggest that large blooms of K. brevis and K. mikimotoi on the West Florida shelf are
not the result of exceptional growth on or the exclusive use of any specific inorganic
or organic P compound, but rather an ability of these species to grow well on a broad
spectrum of naturally occurring DIP and DOP compounds - either through their direct
use (e.g. DIP and phosphate monoesters) or indirectly following bacterial mineralization
(e.g. phosphodiesters and phosphonates). The P requirements of K. brevis and K.
mikimotoi blooms may in part be met by the mineralization of the more refractory
constituents of the DOP pool by the co-occurring bacterial community.
POPULATION STRUCTURE AND GENETIC DYNAMICS OF TOxIC
ALExAndRIum FundyEnSE (DINOPHYCEAE) BLOOMS IN COASTAL
EMBAYMENTS.
Mindy L. Richlen1, Bibiana G. Crespo1,2, Jillian Adams1, Bruce Keafer1, Deana L. Erdner3
and Donald M. Anderson1
Woods Hole Oceanographic Institution, 266 Woods Hole Rd., MS 32, Woods Hole,
Massachusetts 02543, USA
2
Departament de Biología Marina i Oceanografía, Institut de Ciències del Mar, CSIC,
08003 Barcelona, Spain
3
University of Texas Marine Science Institute, Port Aransas, Texas, 78373, USA
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
1
Alexandrium blooms associated with Paralytic Shellfish Poisoning in the northeastern
USA generally occur in two habitats or systems: open coastal waters, spanning hundreds
of kilometers, and coastal embayments, characterized by restricted circulation and
limited tidal flushing. Differences in the environmental and hydrodynamic characteristics
of these systems provide the opportunity for comparative studies of the mechanisms
influencing the genetic structure and diversity of bloom populations. To examine the
effects of estuarine hydrodynamics on the population genetics of Alexandrium fundyense
blooms, we carried out extensive temporal and spatial sampling in the Nauset Marsh
System (NMS) on Cape Cod (MA, USA) in 2009, and again in 2012. Within the NMS, A.
fundyense populations are largely restricted to salt ponds or coves at the distal ends
of the system by hydrographic processes, thus permitting the investigation of how
physical barriers to dispersal influence population substructure of blooms within the
embayments. Temporal sampling was carried out during all phases of the bloom to
determine how the genetic composition of A. fundyense populations changed over time
within a geographically restricted area, when population mixing is limited. These studies
revealed significant spatial differentiation among populations within the NMS, indicating
that population mixing among the terminal endpoints of the system is restricted.
Furthermore, rapid and significant temporal differentiation was observed, with changes
in population structure occurring over the course of the bloom. Results of these
investigations will be discussed, along with the environmental conditions and biological
interactions potentially driving differentiation.
Page 110
CHANS: THE BENEFITS OF PARTICIPATING IN BROADER IMPACTS: A
HIGH SCHOOL TEACHER’S PERSPECTIVE
Katrin Rudge1, Kate Kohler2, Porter Hoagland3, and Barbara Kirkpatrick2,4,5
Riverview High School, Sarasota, FL
3
Marine Policy Center, Woods Hole Oceanographic Institute, Woods Hole, MA
4
Dept of Epidemiology and Public Health, Miller School of Medicine, University of Miami,
Miami, FL
5
Sciences, University of Miami, Miami, FL
1
2
The broader impacts activities of the National Science Foundation Coupled Natural and
Human System (CNH) grant: Collaborative Research: Modeling the Dynamics of Harmful
Algal Blooms, Human Communities, and Policy Choices along the Florida Gulf Coast
include innovative components for engaging with and educating high school students
about our highly cross disciplinary research project.
As an introductory lesson students used technology to create posters about waterborne
diseases in a jigsaw pairing activity with their classmates. Next, students were
introduced to the concept of a dichotomous key with several simplified key examples. A
lab activity incorporated use of a dichotomous key along with microscopic examination
and identification of waterborne disease specimens. Follow-up lessons included a hand
washing, glow germ/black light activity which showed students the importance of
proper hand washing. Once assessed on their knowledge of these diseases a final field
trip was planned to a nearby sewage treatment facility to examine how water is safely
processed and clarified to eliminate chances of spreading waterborne diseases to the
general public.
As a second step in this cooperative learning venture, students were introduced to
the diverse backgrounds of the project investigators in this grant via Safari Live. The
lectures provided students insights into unique career opportunities; varied stages
of careers, from early investigator to emeritus status; differences in institutions from
Academia, to State agencies and non-governmental organizations; incorporating not
only interdisciplinary but also cross disciplinary research examples. Diverse disciplines
students were introduced to included: marine policy, marine biology, engineering, public
health, environmental health, economics, and business/health administration.
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In a cooperative project, Mote Marine scientists and Riverview High School staff,
introduced students the NIEHS “AMBIENT” curriculum, with main focus on the
waterborne disease segment of the curriculum. Below are examples of how students
were engaged and assessed in this teaching unit:
NUTRIENT AND BLOOM DYNAMICS BEFORE DREDGING OF LAKE
NEATAHWANTA, A SHALLOW, FRESHWATER LAKE IN UPSTATE NEW
YORK
Marci L. Savage1 and Gregory L. Boyer2
State University of New York – Environmental Science and Forestry School, 1 Forestry
Drive, NY 13210
2
Drive, NY 13210
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
1
Cyanobacterial species present during bloom conditions can be dictated by the amount
and type of nutrients present. Management plans for shallow lakes include physical
measures such as dredging to remove bottom sediments and reducing the phosphorous
pool. Dredging studies have been published in the past few decades with results varying
from the desired outcome – reduced nutrient levels, increased diversity, decreased
toxin levels, and absence of a bloom, to no significantly discernable differences. Lake
Neatahwanta (Fulton, NY) is one lake that has been plagued by cyanobacterial blooms
seasonally, and this lake is scheduled for dredging in late summer 2013. By monitoring
several parameters including pH, temperature, conductivity, dissolved oxygen, nitrogen,
and phosphorous, a baseline can be prepared for the pre-dredge state of the lake. We
will incorporate these physical parameters along with measurements of algal biomass
and diversity to compare with the post-dredged lake. The change in the species present
affected by this event, and subsequent nutrient changes, will allow a unique opportunity
to model the effect that dredging will have on the species present and toxin levels. This
approach may provide a template that can be applied to other systems for monitor and
predict bloom occurrence and toxicity.
Page 112
VARIATIONS IN THE MICROCYSTIN CONTENT OF DIFFERENT FISH
SPECIES COLLECTED FROM A EUTROPHIC LAKE
Justine R. Schmidt 1, Mylynda Shaskus 2, John F. Estenik 2, Carl Oesch 3, Roman Khidekel 3
and Gregory L. Boyer 1
Department of Chemistry, State University of New York, College of Environmental
Science and Forestry, Syracuse, NY 13210, USA
2
Ohio EPA, Division of Surface Water, Lazarus Government Center, P.O. Box 1049, 50
West Town Street, Suite 700, Columbus, OH 43216, USA;
3
Ohio EPA, Division of Environmental Services (Laboratory), 8955 East Main Street,
Reynoldsburg, OH 43068, USA
1
Page 113
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SPEED ORAL
TALK PRESENTATIONS
Microcystins produced from cyanobacteria can accumulate in fish tissues. Liquid
chromatography coupled with tandem quadrupole mass spectrometry (LC-MS/MS) is
an attractive alternative to immunoassays for the determination of low concentrations
of microcystins in tissues. Fish taken from Grand Lake St. Marys, a eutrophic lake in
Ohio, USA, were analyzed for microcystin-LR in their fillets using LC-MS/MS. Of 129 fish
tested for microcystins, only black crappie (Pomoxis nigromaculatus) and common carp
(Cyprinus carpio) tested positive for microcystin-LR. Less than 10% of Pomoxis and 7%
of Cyprinus samples contained measurable levels of microcystin-LR. Statistical analysis
yielded a p-value of 0.07 between Pomoxis and the pooled results of the other four fish
species. However, this comparison was complicated by the large difference in sample
size between species. Further sampling in Grand Lake St. Marys for microcystin-LR would
help determine if microcystin-LR exposure occurs through foodweb transfer.
ROLE OF AMMONIUM IN ALExAndRIum FundyEnSE BLOOMS IN THE
GULF OF MAINE & GEORGES BANK
Sugandha Shankar, David. W. Townsend, Maura. A. Thomas
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
University of Maine, School of Marine Sciences, 5706 Aubert Hall, Orono, ME 04469,
USA
We present the results of a study designed to test the hypothesis that ammonium is the
more important form of dissolved inorganic nitrogen supporting annual summertime
blooms of the PSP-producing dinoflagellate Alexandrium fundyense in the Gulf of
Maine – Georges Bank region. It is already known that compared to nitrate and nitrite,
ammonium is the more preferred nitrogenous nutrient for phytoplankton because of
its chemically reduced state. Previous oceanographic surveys in the study region have
shown that deep-water nitrate injections into surface waters of the eastern Gulf of
Maine and on the Northern Flank of Georges Bank lead to diatom blooms some distance
downstream in the residual flow field, followed by dinoflagellate blooms, including
populations of A. fundyense. These A. fundyense populations continue to grow and
to maintain high cell concentrations after nitrate is depleted in the surface waters.
Rapid remineralization of particulate nitrogen to ammonium and strong coupling with
subsequent uptake typically keeps ambient ammonium concentrations low; nonetheless,
even very low ammonium concentrations can sustain phytoplankton production.
Results of laboratory culture experiments and multivariate statistical analyses of
field measurements of distributions of A. fundyense and planktonic heterotrophs are
presented, which together support the contention that heterotrophic regeneration of
ammonium is sufficient to sustain continued growth during summertime blooms of A.
fundyense in the Gulf of Maine and on Georges Bank.
Page 114
TRANSCRIPTOME ANALYSIS OF THE DIATOM PSEudO-nITZSCHIA
AuSTRALIS REVEALS PATHWAYS ASSOCIATED WITH DOMOIC ACID
BIOSYNTHESIS
G. Jason Smith1, Thomas J. Savage2, Raphael Kudela3, Kendra Hayashi3 and Holly A.
Bowers4
Moss Landing Marine Laboratories, 8272 Moss Landing Rd, Moss Landing, CA 95039,
USA
2
Department of Chemistry, California State University Sacramento, 6000 J Street,
Sacramento, CA 95819
3
Cruz, CA 95064, USA
4
Monterey Bay Aquarium Research Institute, 7700 Sandholdt Road, Moss Landing, CA
95039, USA
1
DNA-free RNA was submitted to the National Genome Resource Center for RNAseq library construction, sequencing and annotation as part of the Marine Microbial
Eukaryote Transcriptome Sequencing Project (Project IDs: MMETSP_139_2, 140_2,
141_2, 142_2). An average of 23,317 contigs (2178 bp median length) were machine
assembled from each library with 80% having defined protein coding sequences. Of
these putative genes an average of 3899 (22%) sequences per library had significant
homologies (E<10-10) to known proteins enabling pathway reconstruction. P. australis
10249_10AB expressed transcripts encoding enzymes required for complete mevalonate
(hmgr-dependent) and methyl-erithrytol phosphate (dxs-dependent) pathways for
isoprene generation. RNA-seq enables use of normalized read frequency (RPKM) as
an index of relative transcript abundance. Compared to hmgr, dxs abundance was
strongly correlated with DA accumulation (r=0.96) indicating the MEP pathway as the
likely source of the isoprene group on DA. Involvement of the ornithine cycle in supply
of substrates for DA biosynthesis is again indicated by correlation of argD abundance
with DA (r=0.90). Combined with physiological data, this transcriptome database points
to the power of RNA-seq analysis for metabolic network reconstruction and analysis of
toxin biosynthesis.
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SPEED ORAL
TALK PRESENTATIONS
Numerous physiological studies have pointed to a dependence of domoic acid
biosynthesis on the growth status of Pseudo-nitzschia spp, such that DA accu-mulation
is generally enhanced during growth limitation by a range of stressors. While DA
accumulation can be affected through manipulation of growth rate in this diatom genus,
such manipulations confound interpretation of DA biosynthetic patterns and pathway
reconstruction. Here we report on transcriptomes generated from a Monterey Bay
isolate of P. australis (10249_10AB) grown under different N-source supplementations
(NO3, NH4, UREa, GLUtamate) at a common dilution rate. Batch subcultures of antibiotic
treated 10249_10AB were conditioned for 6 passages (ca. 2 months) with specific N
supplements at 40 μM-N and f/2 levels of other constituents. Log phase pre-conditioned
cultures were used to establish continuous cultures under constant light at 15oC with
the same N supplementation supplied at 0.27/d. Cultures were sampled after 10 days
of growth. Total DA accumulation (pg/cell) was highest in the GLU culture (0.253) and
lowest under URE supplementation (0.100) with a rank order of GLU>NO3>NH4>URE.
PURIFICATION AND CHARACTERIZATION OF INDOLE ALKALOIDS FROM A
CENTRAL FLORIDA ISOLATE OF THE FRESHWATER CYANOBACTERIUM,
FISCHERELLA SP., AS INHIBITORS OF VERTEBRATE DEVELOPMENT IN
THE ZEBRAFISH (dAnIO RERIO) EMBRYO MODEL
Dalton D. Steele, Asha Jaja-Chimedza and John P. Berry
SPEEDPRESENTATIONS
TALK PRESENTATIONS ORAL
International University, 3000 N.E. 151st Street, North Miami, FL 33181
Cyanobacteria, also known as the “blue-green algae”, are recognized to produce a
vast array of toxic or otherwise bioactive secondary metabolites. As part of on-going
research focused on identifying bioactive metabolites from cyanobacteria, particularly
as naturally occurring environmental toxicants, extracts of freshwater cyanobacterial
isolates from South and Central Florida were previously screened using the zebrafish
(Danio rerio) embryo as a model of vertebrate development. Using this approach,
extracts from biomass of Fischerella 52-1, a strain of freshwater cyanobacteria
isolated from Central Florida, were found to contain so-called “developmental toxins”
that inhibit or impair pathways or processes in the developing embryo. The present
study focused on using the observed developmental effects, as a means of bioassayguided fractionation, towards purification of relevant metabolites. Recognizing the
production of a diverse repertoire of bioactive indole alkaloids by Fischerella and related
genera, we additionally employed Ehrlich’s spray reagent, as a test for the presence
of indoles, as well as characteristic UV spectroscopic absorbance associated with the
indole chromophore, to specifically target the indole alkaloids as likely contributors
to the observed developmental toxicity. Fractionation was performed using silica-gel
column chromatograpy, and subsequent preparatory thin-layer chromatography (TLC),
followed by purification using reverse-phase high performance liquid chromatography
(HPLC). Indole-containing compounds from the bioactive fractions purified in this way
are currently being characterized chemically by spectroscopic techniques (i.e. mass
spectrometry, NMR, IR), and toxicologically in the zebrafish embryo model. Based on
initial characterization data, one of the purified bioactive compounds is suspected to
be a hapalindole. Further progress in purification, toxicity evaluations and structure
elucidation of relevant metabolites will be reported.
Page 116
HIGH CO2 PROMOTES THE PRODUCTION OF PARALYTIC SHELLFISH
POISONING TOxINS BY ALExANDRIUM CATENELLA FROM SOUTHERN
CALIFORNIA WATERS
Avery O. Tatters1, Leanne J. Flewelling2, Feixue Fu1, April A. Granholm2, David A. Hutchins1
Department of Biological Sciences, University of Southern California, 3616 Trousdale
Parkway, Los Angeles, CA 90089
2
100 8th Ave. SE, St. Petersburg, FL 33701
1
Page 117
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SPEED ORAL
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Marine dinoflagellates are important constituents of microalgal communities in coastal
oceans worldwide. A fraction of them are able to biosynthesize deleterious chemicals
that are capable of impacting human and environmental health. As anthropogenic
CO2 emissions continue to escalate, drawdown of this gas into seawater is leading
to a progressively lower mean pH, referred to as ocean acidification. Concurrently,
increases in sea surface temperatures are being recorded and are projected to continue.
Despite the recognized importance of toxic dinoflagellates, their response to these
climate/global change variables has been relatively unexplored. A clone of paralytic
shellfish poisoning toxin producing Alexandrium catenella (A-11c) was conditioned to
combinations of temperature, CO2 and phosphate concentrations for a period of eight
months. These variables influenced growth and carbon fixation rates and although
these variations only elicited minor differences in toxin composition (on a % basis) or
ratios, the effect on total cellular toxicity was dramatically altered in all treatments.
Cells conditioned to the high CO2, low phosphate at low temperature were the most
toxic and compared to the low temperature of 15 °C, the warmer 19 °C alleviated some
of this toxicity. Overall increased CO2 generally led to enhanced potency. Paralytic
shellfish poisoning toxins are able to vector throughout marine food webs, thus posing
a widespread potential risk to humans, marine inhabitants and their environments.
To our knowledge, this is the first report of how A. catenella may respond to future
marine conditions in Central/Southern California waters. Based on these results further
investigation is warranted.
PHYSICAL MECHANISM FOR KAREnIA bREvIS BLOOM INITIATION IN
TExAS
Kristen M. Thyng1, Robert D. Hetland1, Marcus T. Ogle, Xiaoqian Zhang1, Fei Chen, Lisa
Campbell1
Department of Oceanography, MS 3146, Texas A&M University, College Station, Texas
77843-3146
SPEEDPRESENTATIONS
TALK PRESENTATIONS
ORAL
1
Karenia brevis blooms are known to occur sporadically in Fall, about once every 2-5
years, along the Texas coast. Circulation in the northwestern Gulf of Mexico is suspected
to underlie K. brevis bloom initiation along the Texas coast, and control interannual
variability in bloom presence or absence. This circulation, primarily caused by prevailing
seasonal winds, is investigated using simulated surface drifters in a numerical model run
in the Regional Ocean Modeling System (ROMS). Drifters are moved both forward and
backward in time to understand the movement and origination of drifters for years in
which harmful algal blooms did and did not occur. Bloom occurrence is correlated with
weaker mean downcoast winds in September, as found using station wind data, and with
the presence of southern waters near the Texas coastline, as seen through numerical
drifter experiments. Further understanding of the basic mechanisms of bloom initiation
may lead to better forecasting of these events in the future.
Page 118
QUANTIFYING CYANOBACTERIA AND HIGH BIOMASS BLOOMS FROM
SATELLITE
Michelle C. Tomlinson1, Danielle Dupuy2, Dan Canfield3, Mark Hoyer3, John Hendrickson4,
Rolland Fulton4, Robert Burks4, Erich Marzolf4, Travis Briggs1, Richard P. Stumpf1
CSS-Dynamic, Fairfax, VA 22030, USA
3
University of Florida, Gainsville, FL 32653, USA
4
St. Johns River Water Management District, Palatka, FL 32178, USA
1
2
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PRESENTATIONS
SPEED ORAL
TALK PRESENTATIONS
Algal blooms of high biomass and cyanobacteria are on the rise, occuring both nationally
and internationally. These blooms can foul beaches, clog water intakes, produce toxins
that can contaminate drinking water, and pose a threat to human and domestic animal
health. A quantitative tool can aid in the management need to respond to these issues.
These blooms can affect many lakes within a state management district, pointing to the
need for a synoptic and timely assessment. The 300 m Medium Resolution Imaging
Spectometer (MERIS) satellite imagery provided by the European Space Agency from
2002 to 2012 has led to advances in our ability to monitor these systems. Algorithms
specific to quantifying high biomass blooms are being developed for use by state
managers through a comparison of field radiometry, water quality and cell enumeration
measurements, and remotely-sensed satellite data. Initial evaluations were conducted
for Florida lakes and the St. Johns River. These algorithms are designed to be detect
blooms even with atmospheric interference and suspended sediments. Algorithm
tuning is being evaluated against tunings made for other areas, such as Lake Erie. The
results show that cyanobacteria blooms, especially of Microcystis, can be identified and
their biomass can be estimated (as chlorophyll concentration and other metrics).
THE FOOD QUALITY AND DISSOLVED INORGANIC NUTRIENT EFFECT ON
THE GROWTH AND TOxICITY OF dInOPHySIS ACumInATA FROM NORTH
AMERICA
Mengmeng Tong1,2, Juliette L. Smith2, David M. Kulis2, Donald M. Anderson2
Department of Ocean Science and Engineering, Zhejiang University, Hangzhou, China,
310007
2
Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA
SPEED
TALK PRESENTATIONS
ORAL PRESENTATIONS
1
Dinophysis acuminata, a producer of diarrhetic shellfish poisoning (DSP) toxins and
pectenotoxins, is a mixotrophic species. The ciliate prey and light are required for the
physiological activity of Dinophysis. However, how food quality (i.e. particulate nutrient)
or dissolved inorganic nutrient affects the metabolism of Dinophysis is still unknown.
Here we investigated the growth and DSP toxin production of D. acuminata under two
nutritional sources: food quality and ambient dissolved inorganic nutrient on three
nutritional levels: nutrient replete (N/P=17.3), phosphate limited (N/P=47.4) and
nitrogen limited (N/P=7.0) conditions. Phosphate limitation of prey was accomplished in
the study. The growth rate and cell toxin content of okadaic acid (OA), dinophysistoxin-1
(DTX1) and pectenotoxin-2 (PTX2) did not show any difference in all treatments, with
the cellular OA, DTX1 and PTX2 yielded from 0.18~0.58, 2.2~8.8 and 6.4~15.3pg/cell,
respectively. Toxin was produced at exponential phase. No difference was found in net
toxin production rates in the three nutritional treatments. However, the total toxin
quotas (ng/mL) were significant higher in P-limited treatment when cell aged (at late
plateau phase), with the maximum OA, DTX1 and PTX2 quotas of 9.3±0.5, 91.0±22.7
and 74.0±9.6ng/mL respectively, than that in other two treatments. The higher total
toxin content was driven by the higher biomass of D.acuminata in P-limited condition
(4293cells/mL VS 3986 and 3357cells/mL in nutrient replete and N-limited), which in
return, was driven by the higher biomass of ciliate prey in the mixed culture (1355cells/
mL VS 1078 and 1035cells/mL in nutrient replete and N-limited), indicating the indirect
effect of ambient phosphate substances on the total DSP toxin concentrations in the
culture even no dissolved inorganic nutrient was required by Dinophysis. These results
supported us important information on nutrient availability to dynamic of Dinophysis
blooms and DSP outbreaks in the field.
Page 120
ESTUARINE INFLUENCE ON THE PHYTOPLANKTON COMMUNITY
STRUCTURE DURING A KAREnIA bREvIS BLOOM ON THE FLORIDA GULF
COAST
Jennifer M. Vreeland1, Vincent J. Lovko1, Gary J. Kirkpatrick1, Ari Nissanka1, Val Palubok1,
Susan Launay1, Kellie L. Dixon1
1
Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL, 34236, USA
Page 121
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TALK PRESENTATIONS
In 2012-2013 the southwest coast of Florida experienced a significant bloom of the ‘red
tide’ dinoflagellate Karenia brevis causing massive fish kills, record manatee deaths,
and human respiratory impacts in coastal areas. Bloom initiation is considered to occur
in offshore oligotrophic waters, but less is known about the variety of nutrient sources
that support growth and maintenance as the bloom moves towards the coast. The
estuaries of the central west Florida coast, prominent sources of nutrients to coastal
waters, differ widely in their physical, hydrological and chemical parameters. The scope
of this research was to observe how nutrient levels affect phytoplankton communities in
relation to the development and strength of Florida ‘red tide’ blooms. Throughout the
progression and entirety of the most recent bloom, Mote Marine Laboratory conducted
six nearshore -surveys along the 10 m isobath from Tampa Bay to San Carlos Bay from
August 2012 to April 2013, as part of a cooperative monitoring and research program in
conjunction with the Florida Fish and Wildlife Research Institute (FWRI). Four stations
along this transect were located just outside of the inlets of four major estuarine systems
(Tampa Bay, Sarasota Bay, Charlotte Harbor, San Carlos Bay). Samples collected at these
stations were processed for K. brevis abundance, phytopigments (HPLC), nutrients and
physical water structure. The pigment data were analyzed by chemotaxonomic analysis
(Chemtax) to provide estimated results of the taxonomic structure of the phytoplankton
communities. Primer v6 analyses were used to determine spatial differences and
temporal trends in community structure over the course of the bloom and to associate
community composition (with a focus on K. brevis) and succession with physical and
nutrient concentrations over the duration of the bloom. The statistical relationships
found were interpreted in the context of the effect of nutrients on the maintenance of K.
brevis blooms.
POSTER
PRESENTATIONS
POSTER
ORAL PRESENTATIONS
S
LC-MS METHODOLOGY FOR BIOMARKERS OF BREVETOxINS IN THE
EASTERN OYSTER (CRASSOSTREA vIRGInICA) AND HARD CLAM
(mERCEnARIA SP.) ExPOSED TO KAREnIA bREvIS BLOOMS
Ann Abraham, Edward L.E. Jester, Kathleen R. El Said, Steven M. Plakas and Robert W.
Dickey
ORAL
POSTER
PRESENT
PRESENTATIONS
ATIONS
FDA, Division of Seafood Science and Technology, Gulf Coast Seafood Laboratory,
Neurotoxic shellfish poisoning is caused by the consumption of shellfish contaminated
with brevetoxins. Traditionally, shellfish toxicity is assessed by mouse bioassay. Efficient
in vitro methods are needed for improved monitoring and management programs.
We report the selection of biomarkers of brevetoxin and the development of LC-MS
methodology for applications in regulatory monitoring of brevetoxins in the Eastern
oyster and hard clam. Molluscan shellfish exhibit species-specificity in metabolism
of parent algal brevetoxins when exposed to Karenia brevis blooms. We identified
biomarkers of brevetoxins representative of composite B-type brevetoxins in Eastern
oyster and clam. For oysters, selected biomarkers were the cysteine conjugates BTX-B2
and S-desoxy-BTX-B2. For clams, biomarkers included the cysteine conjugates (BTX-B2
and S-desoxy-BTX-B2), a taurine conjugate BTX-B1, as well as the oxidation product
BTX-B5 and its open A-ring derivative. Oysters and clams collected before, during,
and after K. brevis blooms were analyzed by LC-MS and a commercial ELISA kit. The
composite B-type brevetoxin concentrations as determined by ELISA were highly
correlated with the sum of the biomarker concentrations determined by LC-MS (r2 =
0.94 for both oyster and clam). Comparisons with results of mouse bioassay support the
use of ELISA and LC-MS as rapid screening and determinative/confirmatory methods,
respectively, for monitoring of brevetoxins and assessing toxicity in these species.
Page 124
CHARACTERIZING SENSE AND ANTISENSE RNA POPULATIONS IN
KAREnIA bREvIS AT DIFFERENT TIMES OF THE DIEL CYCLE
Scott B. Anglin1 and Timothy I. McLean1
University of Southern Mississippi, Department of Biological Sciences, 118 College Drive
#5018, Hattiesburg, MS 39406-0001
1
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POSTER
ORAL PRESENTATIONS
Karenia brevis is the responsible agent for generating periodic, if not annual, harmful
algal blooms in certain coastal areas of the Gulf of Mexico. In an effort to better
understand the biology of this organism, a functional genomics project was initiated. As
part of that project, it was determined that a significant number of natural antisense
transcripts (NATs), long non-coding RNAs with reverse complementarity to mRNAs, exist
within the transcriptome of K. brevis. Based on the role of NATs in other organisms,
we hypothesize that K. brevis NATs play a role in regulating gene expression at a posttranscriptional level. To test this prediction, we have extracted total RNA from K. brevis
cells at different points in the diel cycle and sequenced all polyA-containing transcripts
from each sample using the Illumina platform. Bioinformatic analyses will reconstruct
the entire transcriptome of cells at each time point, and then we will cluster, annotate
and determine the relative expression level of each transcript corresponding to the
respective time of day. Specifically, we will assess the relative ratio of each member
of any complementary pairs of mRNA and NAT to determine if the data support or
falsify the hypothesis. The results of these experiments will possibly lead to a better
understanding of environment-gene interactions for this organism, which in turn, will aid
our ability to understand the factors and mechanisms associated with cell growth and
bloom formation.
THINKING ALIKE? COMPARING ExPERT AND PUBLIC PERCEPTIONS OF
THE BENEFITS AND RISKS OF CONSUMING SHELLFISH
Nick Boase1, William Gaze1, Clare Redshaw1,2, Mathew White1, Lora E Fleming1
European Centre for Environment and Human Health, University of Exeter Medical
School, Knowledge Spa, Royal Cornwall Hospital, Truro, Cornwall, TR1 3HD, UK
2
School of Geography, Earth and Environmental Sciences, University of Plymouth, Drake
Circus, Plymouth, Devon, PL4 8AA, UK
1
The shellfish industry is an important part of coastal economies worldwide, including
the UK where shellfish generated 280 million pounds sterling at first point of sale in
2011. Shellfish can also provide a variety of potential health benefits to consumers.
However, as with many food types, shellfish can also present health risks, such as
impacts on human health from consumption of shellfish contaminated with HAB toxins.
UK incidences of shellfish poisoning as a result of HABs are low but consistent, and
harmful algae present in UK waters include Alexandrium spp., Dinophysis spp., and
Pseudo-nitzschia spp. However, little is known about how the UK public perceives the
risks and benefits associated with consuming shellfish, and how this compares with
expert knowledge. Understanding this is important to ensure effective, well-targeted
communication of the risks and benefits from consuming shellfish, resulting in benefits
to both the public and to industry.
POSTER
PRESENTATIONS
ORAL PRESENTATIONS
Aim: This project aims to develop and compare how the public and experts perceive the
risks and benefits (including from HABs) of consuming shellfish, ultimately leading to the
development of effective evidence-based communication about shellfish.
Methods: The well-established Mental Models Approach to Risk Communication, which
establishes people’s perceptions for a specific subject, is used to guide this project.
Semi-structured interviews are underway with experts from across the shellfish supply
chain (e.g. producers) and associated organisations (e.g. government) in the UK, and
also the public. Following analysis of the interview transcripts, influence diagrams will
be constructed to represent the mental models for each of these two groups, and then
compared. In addition, language used by the public and experts to describe risks and
benefits will be discussed.
Results/Conclusions: Findings from initial pilot studies indicated potential
misconceptions amongst the public over some of the risks and benefits from consuming
shellfish; for example, in recognising the difference in potential risk exposure between
self-harvesters and those sourcing from a commercial supply chain. The findings are
expected to inform communications about shellfish between industry and the public to
resolve such misconceptions. In addition, this project will inform a subsequent largescale survey of the UK public to understand the prevalence of these perceptions, and
how these relate to shellfish eating behaviour.
Page 126
LADDER FRAME POLYETHERS AS POTENTIAL DRUG TRANSPORTERS
Andrea Bourdelais1, Elizabeth Elliott1, Karl Jacocks1, Allan Goodman1, Vinita S. Chauan2,
Ian Marriott2 and Daniel G. Baden1
University of North Carolina at Wilmington, Center for Marine Science Wilmington, NC
28409 USA
2
University of North Carolina at Charlotte, Department of Biology, Charlotte, NC 28223 USA
1
Ladder framed polyethers (LFPs) are a class of naturally occurring compounds that are
produced by a variety of marine dinoflagellates. Many of the LFPs are highly toxic and
most show selective binding to receptor sites on ion channels. Examples of LFPs include;
brevetoxins, hemibrevetoxin B, brevenal, brevisin, ciguatoxins, maitotoxins, gambierols
and yessotoxins.
Brevetoxin’s innate toxicity would likely preclude it from being suitable for drug delivery,
so BODIPY-LFP conjugates were also prepared for brevenal and brevisin and studied
for a similar ability to act as drug transporters. These smaller, less toxic, LFPs transport
membrane impermeable compounds into cells as quickly and as efficiently as brevetoxin.
In addition to the previous experiments we have tested the BODIPY and 6-TAMRALFP conjugates in 9 different cell lines, determined the EC50 in cytotoxicity assays and
determined subcellular localization. Additionally, BODIPY-brevisin was administered to
mice and whole body distribution determined. The results from these experiments will
be presented.
Page 127
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October 2013
ORAL PRESENTATIONS
POSTER
Perhaps the most well characterized LFPs originate from the marine dinoflagellate
Karenia brevis and include the brevetoxins, which has been shown to modulate voltage
sensitive sodium channels (VSSC’s). Brevetoxins activate VSSC’s and exposure to these
toxins via consumption of contaminated seafood can result in neurotoxic shellfish
poisoning. In an attempt to fully understand the functional activity of the brevetoxins,
development of a simple fluorescence based receptor binding assay for brevetoxin in
living cells was attempted. Fluorescent conjugates of brevetoxin were prepared using a
variety of the fluorophores (BODIPY®, Texas red, 6-TAMRA, D355, C356, A10439, D100,
D355, P101, and 4-hydroxybenzhydrizide),. Competitive binding studies using rat brain
synaptosomes showed most of the fluorescent brevetoxin conjugates retained good
affinity for the receptor. High affinity compounds were then tested for their brightness
and fade resistance in SJCRH30 human rhabdomyosarcoma cells. The fluorophore
conjugates of BODIPY, 6-TAMRA, and C356 were very bright and easily visualized in
living cells. C356, a fluorescein derivative proved to fade quickly, therefore, BODIPY
and 6-TAMRA were chosen for further studies. Higher magnification of cells treated
with the fluorophore-brevetoxin conjugates showed that the compounds were not
localized on the cell surface but had been internalized, invalidating the development of a
fluorescence based receptor binding assay. Although their utility for identifying receptor
binding sites in whole cells was limited, the rapid uptake of membrane impermeable
fluorophores when conjugated to brevetoxin led to the hypothesis that brevetoxins
and, possibly, other LFPs may have utility to rapidly transport compounds, with low
membrane permeability, across cell membranes.
INNOVATIVE DRINKING WATER TREATMENT FOR TASTE AND ODOR
REMOVAL WITH NATURALLY OCCURRING BACTERIA FOUND IN SURFACE
WATER
A.J. Boyes, K. Gilmore, B. MacLeod, A. Shawvner-Karnitz, and N. Moran
Manatee County Water Treatment Plant 17915 Waterline Rd. Bradenton FL, 34212 USA
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Taste and odor (T&O) compounds found in surface waters globally are produced
as secondary algal metabolites by certain cyanobacteria. These compounds,
2-methylisobrneol (MIB) and geosmin are tertiary alcohols which impart earthy-musty
tastes and odors during cyanobacterial blooms. These odorants are not removed by
conventional treatment at drinking water facilities, and can be detected in by consumers
at concentrations as low as 5-10 ng L -1. Traditionally, powder activated carbon (PAC) is
used as additional treatment to mitigate the T&O effects but it is a costly process and is
sometimes unsuccessful at removing the offending odorants. Drinking water industries
are facing the need for more innovative and cost effective technologies for water
treatment and purification to remove these compounds.
A bench-scale study initiated in 2006 at the Manatee County Water Treatment Plant
(Bradenton, Florida) and later a pilot-scale study starting in 2009, investigated direct
biological filtration of raw surface water prior to conventional treatment for removal of
these T&O compounds. Results indicated that spent granular activated carbon (GAC)
or anthracite contactors which were allowed to colonize with bacteria from the Lake
Manatee achieved successful removal of the odorants. When applying a spike of 2476
ng L-1 of geosmin to the pilot columns, 99.8% of the odorant was removed in a 12.6
minute empty bed contact time. This value of geosmin spike represents the maximum
concentration in the past 10 years of historical occurrences. Direct biological filtration
was shown to be an effective treatment option for removal of the odorants MIB and
geosmin from warm Florida waters with high TOC content.
Page 128
MITIGATION OF A TOxIC ALExAndRIum BLOOM USING HYDROGEN
PEROxIDE
Amanda Burson1, Hans C.P. Matthijs1, Renee Talens2, Wilco de Bruijne2, Ron
Hoogenboom3, Arjen Gerssen3, Kees Steur4, Yvonne van Scheppingen4, Anne Fortuin4,
Petra M. Visser1, Maayke Stomp1, and Jef Huisman1
Aquatic Microbiology, Institute for Biodiversity and Ecosystem Dynamics, University of
Amsterdam, The Netherlands
2
Arcadis Netherlands
3
RIKILT Wageningen UR, The Netherlands
4
Waterschap Scheldestromen, The Netherlands
1
Page 129
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ORAL PRESENTATIONS
POSTER
The dinoflagellate Alexandrium ostenfeldii is a well known harmful algal bloom (HAB)
species that can cause paralytic shellfish poisoning (PSP). Although detrimental
bloom events of A. ostenfeldii have occurred in Western Europe, the Netherlands
have historically not shared this problem. However, in August of 2012 a dense bloom
occurred in the brackish Ouwerkerkse Kreek (Zeeland) and its connecting channels. The
A. ostenfeldii strain produced both saxitoxins and spirolide toxins which killed a dog
via ingestion. The Ouwerkerkse Kreek routinely discharges water via a pump station
into the adjacent Oosterschelde estuary, and an immediate call for remediation of the
bloom was therefore required to avoid contamination of extensive shellfish grounds.
Previously, treatment of infected waters with hydrogen peroxide (H2O2) successfully
suppressed cyanobacterial blooms in Dutch lakes. Therefore, we adapted this treatment
to fight off the Alexandrium bloom using a three-step approach. First, we investigated
the required H2O2 dosage in laboratory experiments with Alexandrium. Second, we
tested the method in a small isolated channel adjacent to the Ouwerkerkse Kreek.
Finally, we applied the optimized method to the entire Kreek. Vegetative cell numbers
were depleted up to 99% within 48 hours, and toxins in the water were reduced below
the safety requirements of 15 ng/mL. The added H2O2 decayed to water and oxygen
within 72 hours and fish mortalities were minimal. This is the first application of
H2O2 to suppress a marine HAB species. This research provides a possible option for
management of Alexandrium spp. blooms and other HAB events.
LOCALIZATION OF BREVETOxINS AND PHOTOAFFINITY LABELING OF
TARGET PROTEINS
Wei Chen, Ryan Cassell and Kathleen Rein
Department of Chemistry and Biochemistry, Florida International University, Miami, FL
33199
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Karenia brevis, the major harmful algal blooms (HABs) dinoflagellate of the Gulf of
Mexico, plays a destructive role in the region. Because Karenia brevis produces potent
neurotoxins, known as the brevetoxins, it has negative impacts on coastal economies,
marine life and public health. Brevetoxins are ladder-shaped polyether (LSP) compounds.
The brevetoxins have caused widespread marine animal mortalities and human
poisonings. This has been attributed to their affinity for voltage-sensitive sodium ion
channels causing channel opening and depolarization of excitable cell membranes.
The endogenous role of brevetoxins is unknown. However, it was recently proposed
that brevetoxins play an important role in osmoregulation by Karenia brevis. Using
fluorescently labeled brevetoxin and brevetoxin photoaffinity probes, we will attempt
to localize brevetoxin to a subcellular organelle or a target protein. This may implicate a
specific role for the brevetoxins and other PE ladder type molecules.
Page 130
BREVETOxIN METABOLISM AND PHYSIOLOGY – A FRESHWATER MODEL
OF MORBIDITY IN ENDANGERED SEA TURTLES
Courtney Cocilova1, Jennifer Yordy2, Courtney Bennett3, Gregory Bossart4, Leanne
Flewelling5, Catherine Walsh6 and Sarah Milton7
1
Florida Atlantic University, 777 Glades Road, Boca Raton, FL 33431, USA
2
3
4
Georgia Aquarium, 225 Baker Street NW, Atlanta, GA 30313, USA
5
Florida Fish and Wildlife Conservation Commission, 100 Eighth Ave SE, St. Petersburg, FL
33701, USA
6
7
Florida Atlantic University, 777 Glades Road, Boca Raton, FL 33431, USA
Page 131
S
October 2013
ORAL PRESENTATIONS
POSTER
The dinoflagellate Karenia brevis is a key organism in harmful algal blooms (HABS,
Florida Red tide) that occur off the coast of Florida. K. brevis produces a suite of
neurotoxins which are collectively referred to as brevetoxins (PbTx). PbTx-3 is one of the
brevetoxin congeners and is known to bind voltage-gated sodium channels which affect
cellular permeability, resulting in a cascade of events leading to cell death. Brevetoxin
exposure affects marine life by interrupting neurological functions, decreasing immune
function, inducing inflammation and causes respiratory effects in humans. HABs are
increasing in frequency and distribution and are not only of immediate concern to
species that inhabit areas where these blooms occur, but may have long term effects
due to biomagnifications and bioaccumulation. In 2005, at least 109 loggerhead sea
turtles in Florida were affected by red tides with over 70 impacted during a 2006 bloom.
However, brevetoxicosis is difficult to treat in sea turtles as the physiological impacts
have not been investigated and magnitude and duration of brevetoxin exposures, as
well as specific toxin congeners involved are generally unknown. We are using fresh
water turtles (Trachemys scripta) as a model for brevetoxin exposure in marine species.
Turtles were exposed to 3.12 mg/kg PbTx-3 by intratracheal instillation (0.05ug/
ul) every other day for one week and harvested after different times post-exposure
(24h, 48h, 1w, 2w, 1m). Tissues from the heart, kidneys, brain, fat, intestine, liver and
lymphoid tissues including the spleen and lungs were frozen in liquid nitrogen for
ELISA analysis to investigate uptake, tissue distribution and routes of excretion as well
as for histology. Blood samples were collected and used for immune analysis studies.
Short-term exposures resulted in few immunological or pathological changes in T.
scripta, though the data did suggest that severe interstitial pneumonia with edema
may be a specific condition related to PbTx-3 exposure in this species. Blood analysis
did not show any changes in hematocrit or plasma (lysozyme, superoxide dismutase,
or glutathione-S-transferase) enzyme activity. Some immune function parameters
were altered with exposure, including a significant decrease in phagocytosis after 48 h
exposure and reduced lymphocyte proliferation after 1 month exposure. Plasma protein
electrophoresis profiles showed a decrease in plasma albumin and corresponding A:G
after 24 h. Longer exposure times to PbTx-3 will be necessary to accurately determine
which organ systems become most severely impacted. Future studies will include
in vitro and in vivo work to further determine the role of PbTx-3 as well as devising
appropriate treatment plans to implement when such blooms occur.
ASSESSMENT OF THE EASTERN GULF OF MExICO HARMFUL ALGAL
BLOOM OPERATIONAL FORECAST SYSTEM: A COMPARATIVE ANALYSIS
OF FORECAST SKILL AND UTILIZATION, 2004-2012
Edward Davis1, Karen Kavanaugh1, Katherine Derner2 and Cristina Urizar3
Services (CO-OPS), Silver Spring, MD 20910, USA
2
NOAA, National Ocean Service, CO-OPS, 672 Independence Parkway, Chesapeake, VA
23320, USA
3
NOAA, National Ocean Service, CO-OPS, 263 13th Avenue South,
St. Petersburg, Florida 33701, USA
1
POSTER
PRESENTATIONS
TIONS
ORAL
PRESENTA
To aid early bloom identification and response efforts, in 2004 NOAA transitioned a
successful forecast system for harmful algal blooms (HABs) from research to operations
along the Gulf coast of Florida. NOAA’s Gulf of Mexico HAB Operational Forecast
System (HAB-OFS) issues weekly bulletins that serve as decision support tools for
coastal resource managers, federal and state agencies, and academic institutions. In
an ongoing effort to improve the HAB-OFS, bulletin utilization and forecast quality are
evaluated regularly. Utilization is measured by user feedback and observed or reported
mitigating actions from public resource managers in response to bulletin information.
Forecast accuracy, reliability and skill (i.e. relative accuracy) are calculated for each of
the following forecast components: bloom transport, bloom intensification, and the daily
potential for respiratory irritation along the coast.
Blooms of the toxic dinoflagellate Karenia brevis were present alongshore southwest
Florida in the Gulf of Mexico four out of five years during the assessment period from
May 1, 2008 to April 30, 2012, impacting public health, ecosystems, and regional
economies. During this time, over 200 bulletins were issued for the eastern Gulf of
Mexico, covering 4 individual bloom events, totaling 526 bloom days. Preliminary
analysis shows that forecast accuracy each year was highest for respiratory irritation
forecasts, with the highest relative accuracy amongst the “moderate” and “high” level
impacts. However, all ‘very low’ respiratory irritation forecasts were unassessable. These
results highlight challenges faced by the HAB-OFS team. Confirmation of respiratory
irritation relies on a network that does not cover the entire forecast area and therefore
cannot adequately confirm all respiratory forecasts. At least 90% of respiratory irritation
forecasts could not be assessed due to lack of information. Additionally, the ‘very low’
respiratory irritation category is difficult to assess because it corresponds with a level
of irritation that is limited to individuals with chronic respiratory conditions, such as
asthma. These results will be compared to the assessment of previous operational years
(2004 to 2008) and used as guidance to improve forecasting protocols for the HAB-OFS
program in the future.
Page 132
ASSESSMENT OF THE WESTERN GULF OF MExICO HARMFUL ALGAL
BLOOM OPERATIONAL FORECAST SYSTEM: ANALYSIS OF FORECASTS
AND UTILIZATION OVER THE FIRST TWO OPERATIONAL YEARS, 2010-2012
Katherine Derner1, Karen Kavanaugh2, Edward Davis 2, and Cristina Urizar 3
Services (CO-OPS), 672 Independence Parkway, Chesapeake, VA 23320, USA
2
NOAA, National Ocean Service, CO-OPS, Silver Spring, MD 20910, USA
3
NOAA, National Ocean Service, CO-OPS, 263 13th Avenue South, St. Petersburg, FL
33701, USA
1
This analysis details the assessment of bulletin utilization and forecast skill for the
first two years of operational status along the Texas coastline, encompassing 106 total
bulletins issued for the western Gulf of Mexico from October 2010 through April 2012.
Of those, 44 bulletins were issued during the 2011 Texas red tide, one of the longest
lasting and largest blooms on record in Texas, covering the coastline and inshore bays
and waterways from South Padre Island to the Galveston region. Preliminary analysis
for the first two operational years shows a relatively high degree of forecast skill for
respiratory irritation forecasts, with greater than 70% confirmed correct, and greater
than 80% of all moderate and high level forecasts confirmed correct. Heidke skill scores
for respiratory irritation forecasts ranged between >.30 to .90, indicating a >30% to
90% improvement in forecast accuracy when compared to chance. These results also
highlight the special challenges in the assessment of each forecast component. The
majority of respiratory irritation forecasts, which are reliant on daily reports of field
observations, could not be assessed due to a lack of information. The high amount
of resuspension present along the Texas coastline also presents challenges in the
assessment of transport, which is dependent upon the ability to discern and track
distinct features through satellite imagery. A comparison in forecast assessment
between the eastern Gulf of Mexico (Florida) and the western Gulf of Mexico (Texas) is
also explored to highlight possible enhancements in product operations and assessment.
ORAL PRESENTATIONS
POSTER
October 2013
S
Blooms of the toxic dinoflagellate, Karenia brevis, occur nearly every year in coastal
regions of the Gulf of Mexico causing impacts on public health, ecosystems, and regional
economies. Following the successful transition of the eastern Gulf of Mexico forecast
system for harmful algal blooms (HABs) from research to operations along the gulf coast
of Florida in 2004, NOAA expanded its Gulf of Mexico HAB Operational Forecast System
(HAB-OFS) to include the coast of Texas (western Gulf of Mexico) in 2010. NOAA’s Gulf
of Mexico HAB-OFS issues weekly bulletins that serve as decision support tools for
coastal resource managers, federal and state agencies, and academic institutions. The
Texas bulletins include three forecast components: bloom transport direction, transport
distance and the daily potential for respiratory irritation along the coast. In order to
continually improve the HAB-OFS, forecast skill and bulletin utilization are evaluated
regularly. Each of the forecast components is statistically compared to observational
data. Utilization is measured by user responses and observed or reported mitigating
actions based on bulletin information.
Page 133
KAREnIA SP. IN SW FLORIDA – TRAJECTORIES OF NITROGEN, SILICA,
OTHER WATER QUALITY, AND TAxONOMIC COMPONENTS DURING
THREE ExTENDED BLOOMS
Kellie Dixon1, Ari Nissanka1, Jason M. Lenes2, Jennifer M. Vreeland1, and Gary J.
Kirkpatrick1
1
University of South Florida, 140 7th Avenue South, St. Petersburg, FL 33701, USA
ORAL
POSTER
PRESENTATIONS
PRESENTATIONS
2
Three harmful algal blooms of Karenia brevis in the coastal waters of SW Florida were
investigated for behavior over time of K. brevis and the biomass of other phytoplankton
taxonomic groups, with the progressions statistically related to a variety nutrient
and physical parameters, and with a particular emphasis on available nitrogen and
silica supplies. Allocation of chlorophyll a among the various taxa (K. brevis, diatoms,
haptophytes, chlorophytes, cyanophytes, prochlorophytes, cryptophytes, and other
dinoflagellates) was performed via HPLC and Chemtax analyses. Available data were
limited to coastal samples with K. brevis cell counts greater than 2000 cells L-1 to
emphasize bloom growth and maintenance phases rather than non-bloom and bloom
initiation phases. The blooms evaluated represented a range of conditions; two more
‘typical’ occurrences beginning in late summer through early winter (2006, 2012-2013),
and a less common occurrence initiating during the winter of 2005. Some taxonomic
groups (cyanophytes and prochlorophytes) displayed reproducible seasonal groupings,
while others varied over time but in less correlation with thermal cues. At times, K.
brevis biomass comprised nearly 90% of all phytoplankton, predominantly displacing
diatoms, cyanophytes, and chlorophytes. In a monthly progression for each bloom, K.
brevis dominance was reduced as DIN:DSi ratios increased from <0.3 to >0.5 μM:μM,
although the replacement species was not always diatoms. A multivariate analysis
(Primer V6) determined that samples were distinguished predominantly by biomass
due to K. brevis, diatoms, and followed to a much lesser extent by cryptophytes and
chlorophytes. Community similarity among samples (Bray-Curtis) was significantly
associated with selected nutrients and physical parameters, with results varying among
blooms. Significant explanatory parameters variously included temperature, inorganic
phosphorus, silica, and ammonia concentrations.
Page 134
A PERSISTENT BLOOM OF AnAdyOmEnE J.V. LAMOUROUx
(ANADYOMENACEAE, CHLOROPHYTA) IN BISCAYNE BAY, FLORIDA.
Ligia Collado-Vides1, Natalie Dou1 Christian Avila2, Stephen Blair2, Frederik Leliaert3; Dení
Rodriguez4, Sabrina Schneider1, Pamela Sweeney5, Diego Lirman6
. Florida International University, Miami, FL 33199, US
DERM Miami-Dade County, FL 33136, US
3.
Ghent University, Krijgslaan 281 S8, 9000, Ghent, Belgium
4.
Univerisdad Nacional Autónoma de México. D.F. 04510, México
5.
Department of Environmental Protection, Miami, FL 33138, US
6.
University of Miami, 4600 Rickenbacker Cswy., Miami, FL 33149, US
1
2.
Page 135
S
October 2013
POSTER
ORAL PRESENTATIONS
Green macroalgal blooms are becoming a common problem in coastal waters and
estuaries. This study describes the first occurrence of a persistent macroalgal bloom of
the genus Anadyomene J.V. Lamouroux (Cladophorales, Anadyomenaceae) in the world
and particularly in Biscayne Bay, Florida, USA. The morphological-based identification
of species was verified by a molecular analysis that sequenced the variable C1D2
region of the large subunit (LSU) nrDNA. Results indicate that the bloom is composed
of two species: Anadyomene stellata, reported previously for Florida, and a diminutive
perforate undetermined species, Anadyomene. sp. Morphological traits of Anadyomene
sp. are similar to Anadyomene linkiana, however the lack of vein-encircled perforations
and molecular data of the original species impede us to give a definitive identification
to these species. However it is potentially representing an introduction in the area.
General surveys in Biscayne Bay based on a stratified random design, to visually estimate
the percent cover of submerged aquatic vegetation, date from 1999, using the same
methods recent intensive surveys of the detected bloom were conducted once a year
from 2010 to 2012. Results show that the Anadyomene bloom densities have persisted
since 2005 through 2012 covering an area of approximately 60 km2 of seagrass habitats.
The spatial distribution of the bloom is restricted to the central inshore section of the
Bay, an area affected by canals and groundwater discharges. The persistent 75 % cover
reported for several sites, has caused significant negative impacts to seagrass beds. This
bloom occurring adjacent to metropolitan Miami, adds to the world trend of increasing
green macroalgal blooms occurring at enriched coastal waters.
A SYNOPTIC VIEW OF FLORIDA INLAND ALGAL BLOOMS
Danielle Dupuy1, Michelle C. Tomlinson2, Rolland Fulton3, John Hendrickson3, Robert
Burks3, Erich Marzolf3, Travis Briggs2, Richard P. Stumpf2
CSS-Dynamic, Fairfax, VA 22030, USA
1
2
St Johns River Water Management District , Palatka, FL 32178, USA
3
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Toxic cyanobacterial blooms have become an increasing occurrence in Florida’s lakes,
rivers, and estuaries. Cyanobacterial blooms pose a threat to human and animal
health and degrade water quality, which can have substantial effects on aesthetic,
ecological, and recreational values. Expansive monitoring is essential for the successful
management of these resources. High resolution remote sensing products, like the
300m ocean color products available from the medium-spectral resolution imaging
spectrometer (MERIS) have shown success in capturing Florida’s smaller water bodies.
These remotely-sensed products and in situ data acquired from various sources, which
include Florida LAKEWATCH and St Johns River Water Management District (SJRWMD),
provide a comprehensive view of Florida’s inland algal blooms from 2009 -2011.
Seasonal and interannual patterns in bloom activity can be tracked throughout the
state, as well as the development of individual bloom events, such as the cyanobacterial
bloom in the St Johns River in 2011. This information is a useful supplement to current
agency monitoring programs for directing field sampling and management efforts.
Page 136
UPTAKE MECHANISM OF FLUORESCENT CONJUGATES OF KAREnIA
bREvIS LADDER FRAME POLYETHER COMPOUNDS
Elizabeth A. Elliott, Allan Goodman, Daniel Baden and Andrea Bourdelais
Center for Marine Science, UNCW, 5600 Marvin K. Moss Ln, Wilmington, NC 28409
The marine dinoflagellate Karenia brevisproduces a host of toxins (brevetoxins) and nontoxic natural compounds (brevenal and brevisin) classified as ladder frame polyethers
(LFPs).In an attempt to develop a fluorescentreagent for use in the develop-ment of a
whole cell receptor binding assay, a series of LFP-fluorophore conjugates were prepared
using cell impermeable fluorescent compounds. However, when cells were treated
with these conjugates, it was found that the compounds were rapidly transported into
the cell. Based on the chemical structure of the fluorophore, the LFP conjugates were
internalized to different organelles within the cell. The ability for the LFPs to facilitate
the uptake of cell impermeable compounds has led us to dub them as Escortins.
To discern which active transport mechanism the LFP-fluorophores were utilizing,
a series of experiments were performed using selective uptakeinhibitors. From
this, it was determined that brevetoxin, brevenal and brevisin all enter the cell via
dynamin dependent mechanisms. However, using more specific inhibitors, it has been
discovered that all three LFPs utilize slightly different pathways within the dynamin
mediated mechanism. Results show that brevetoxin and brevisin are facilitated by
caveolin dependent mechanisms, but appear to have subtle differences in their specific
pathways, whereas brevenal appears to utilize a separate mechanism. These findings
suggest that the different LFP conjugates may offer the tools necessary to better
understand these different uptake mechanisms. A complete description of methods and
results from these studies will be presented.
ORAL PRESENTATIONS
POSTER
October 2013
S
Studies were initiated to determine whether the conjugates were taken up into
the cellsvia simple passive diffusion or through active transport mechanisms.Live
SJCRH30cells at either 4 °C or 37 °C were treated with LFP-fluorophore conjugate and
the uptake of the compound was measured using live cell imaging andfluorescence
intensity scoring software.Results showed a significant reduction in uptake at the
lower temperature, suggesting that uptake was not through passive diffusion. To
further confirm this, studies utilizing 2-deoxy-D-glucose to reduce all active transport
mechanisms were performed. In this study, cells werepretreated with the deoxyglucose
to deplete the cells of an energy source. The cells werethen treated with the LFPfluorophore conjugates and the uptake measured as before. Again, comparison of
these results with controls grown under normal conditions, showed a decrease in LFPfluorophore uptake, further supporting an active transport mechanism.
Page 137
MONITORING STRATEGIES FOR THE CIGUATERA-CAUSING BENTHIC
DINOFLAGELLATE, GAmbIERdISCuS
Amanda Ellsworth, Ashley Brandt and Michael L. Parsons
Florida Gulf Coast University, 10501 FGCU Boulevard South, Fort Myers, FL 33965, USA
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Ciguatera is the leading form of phycotoxin-related seafood poisoning in humans
worldwide. Ciguatera is caused toxins produced by some members of the benthic,
epiphytic dinoflagellate genus, Gambierdiscus. Bioaccumulation of these toxins in coral
reef food chains ultimately affects human health and food seafood safety.
Gambierdiscus has been shown to exhibit preferences for some host macrophytes (e.g.,
filamentous rhodophytes) over others (e.g., chlorophytes). While such preferences may
reflect differences in chemical attraction/revulsion between different host species, the
difference in preference may simply be a function of surface area (i.e., a filamentous
thallus will have more surface area than a blade). Differences in host preference,
coupled with a research need to compare Gambierdiscus populations between different
sites and regions (where different host species are likely to be present), have led to
efforts to use neutral (artificial) substrates as a proxy for host macrophytes. In this
study, three different artificial substrates (burlap, nylon screen, and PVC tiles) were
compared at four sites in the Florida Keys, in the vicinity of Long Key. The substrates
were deployed monthly over the course of one year to determine which substrate
was best representative of the predominate macrophytes indigenous to those areas.
Results demonstrated that Gambierdiscus cell densities from the PVC tiles were better
correlated with the Gambierdiscus cell densities on the macrophytes than the other
substrates. For this reason, coupled with their reliability, durability, and ease of use,
tiles were chosen over the other two artificial substrates for the continued monitoring
of Gambierdiscus populations around Long Key. Monthly monitoring of Gambierdiscus
populations using macrophytes and artificial substrates showed seasonal variation,
possible causes of which will be discussed.
Page 138
NATURAL MORTALITY DURING THE DECLINE OF ALExAndRIum BLOOMS
C.J. Choi and D.L. Erdner
The University of Texas at Austin, Marine Science Institute, Port Aransas, TX 78373 USA
While our knowledge of factors controlling the development of phytoplankton blooms
is increasing, the mechanisms leading to bloom decline and termination are not well
characterized. It has been assumed that grazing and sedimentation can cause a decline
of phytoplankton populations, and more recently, lytic viruses and programmed cell
death (PCD) have been recognized as sources of mortality. However, the relative
significance of the various mechanisms is not well understood. To distinguish factors
regulating bloom decline, we studied blooms of Alexandrium spp. in Mill Pond and Salt
Pond within the Nauset Marsh System (Cape Cod, MA) where this species forms massive
blooms annually. Weekly samples were collected at four depths (1m, 3m, 5m, and
7m) from March to May 2013, to investigate the spatial and temporal extent of bloom
initiation, development, and decline. We focused specifically on mortality caused by
PCD as a factor in bloom decline. A greater than 10% increase in dead Alexandrium cells
was observed during the bloom decline, highlighting the potential for natural mortality
to contribute to Alexandrium bloom termination. In addition to quantifying PCD, we
investigated the role of reactive oxygen species (ROS), known mediators for PCD. No
substantial cell death was observed during the peak of the blooms, and evidence of
other potential factors for bloom demise, e.g. induction of sexuality was observed. The
contribution of these different mechanisms to bloom termination will be discussed.
ORAL PRESENTATIONS
POSTER
Page 139
S
October 2013
THE EFFECT OF TEMPERATURE CONDITIONING ON ALExAndRIum
FundyEnSE CYST GERMINATION DYNAMICS IN A SHALLOW ESTUARINE
SYSTEM
Alexis D. Fischer1 and Donald M. Anderson1
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA
The flux of cells from germinated cysts is critical to the population dynamics of many
harmful algal bloom species. A distinctive feature of the widespread, open coastal
blooms of Alexandrium fundyense in the Gulf of Maine region is an endogenous annual
clock that restricts cyst germination to a discrete interval in the spring and early summer.
Relatively little is known of the mechanism regulating the seasonality of excystment of
A. fundyense from shallow water habitats. Through cyst germination experiments, the
control of seasonal excystment was examined in the Nauset Marsh System (NMS), a
shallow estuary on Cape Cod, Massachusetts (USA) with recurrent blooms tied to cyst
seedbeds. No endogenous clock was evident in the germination of cysts produced in
the previous spring bloom and stored under cold, dark, anoxic conditions for 18 months.
To investigate seasonal temperature regulation, freshly sampled cysts from the NMS
were isolated monthly into well plates and incubated at the ambient temperature, with
excystment monitored weekly. Regardless of the incubation temperature and month
isolated, 68(±6)% of the cysts had germinated after 120 days. Excystment success after
30 days of incubation had an autumn minima and spring maxima. Maximum values
were coincident with the water temperature increasing from its annual minimum in
January. Minimum values were coincident with the water temperature decreasing
from its annual maximum in August. Seasonal excystment patterns were independent
of a temperature function. Although the same ambient temperature (2°C) was used
to incubate both January and February cyst populations, the January cysts required
more than twice as many days of incubation to achieve 10% germination, yet the same
length of incubation was required by both populations to attain 50% excystment. This
lag experienced by the January cysts can be explained by the additional month of field
conditioning the February cysts experienced. Excystment success of A. fundyense in
the NMS seems to be modulated by a history of the seasonal temperature patterns.
Like the Alexandrium cysts of Cork Harbor, Ireland and the seeds of higher plants,
secondary dormancy is likely a strategy used by A. fundyense cysts in the NMS to prevent
germination during conditions unfavorable for growth. With secondary dormancy, cyst
germination timing is more flexible than is the case with endogenous clock regulation,
and thus species with this strategy would be best suited for shallow water systems
where interannual variation in environmental conditions are significant. This strategy
would also allow the species to be more adaptable to climate-related environmental
fluctuations.
Page 140
A MODIFIED CONCEPTUAL MODEL INTEGRATING HEALTH AND
ENVIRONMENTAL IMPACT ASSESSMENT: HARMFUL ALGAL BLOOMS
Lora E. Fleming1, Stefan Reis2, George Morris1, Sheila Beck3, Michael H. Depledge1,
Melanie Austen4, Mathew White1, Tim Taylor1
School, Knowledge Spa, Royal Cornwall Hospital, Truro Cornwall, TR1 3HD, UK
2
Centre for Ecology & Hydrology, Bush Estate, Penicuik, EH26 0QB, UK
3
NHS Health Scotland, Meridian Court, 5 Cadogan Street, Glasgow G2 6QE UK
4
Plymouth Marine Laboratory, Prospect Place, The Hoe, Plymouth, PL1 3DH, UK
1
Background - Methodologies for risk and impact assessment of human and
environmental health effects often incorporate the use of conceptual models for
problem framing and for the identification and evaluation of interventions. As
the understanding of the intricate relationships between human health and the
environment grows - particularly with regard to humans as part of ecosystems - the
need for integration across scientific and professional disciplines and policy communities
increases.
Aims - Through linking environmental health and ecosystem services, we aim to
address inefficiencies introduced by discipline and policy barriers and so foster mutual
understanding and improvements to human health and well-being.
Conclusions - Using the example of harmful algal blooms, we highlight the need to
account for second order effects on human health via ecosystem services. Furthermore,
we show socio-economic context is vital to account for distributional effects and
modifiers for exposure-effect-relationships. We also recognise the potential for
positive health and well-being effects (e.g. through green or blue space; or blue carbon
initiatives). Quantifications of effects which omit these positive aspects are potentially
misleading, wrongly implying an overall reduction in societal welfare. Also it is vital to
consider where measures to address an issue may create new problems.
Page 141
S
October 2013
ORAL PRESENTATIONS
POSTER
Methods/Results - We propose the “Ecosystem-Enriched Driver-Pressure-StateExposure-Effect (eDPSEEA) model,” to overcome the fragmentation of scientific
disciplines and policy areas. It recognises positive and negative connections between
the health of both humans and the environment. eDPSEEA incorporates socio-economic
and other contextual modifiers of exposure and exposure-effect relationships, and
recognises the importance of feedback-loops, trade-offs, and synergies between actions.
As an illustration, we discuss how harmful algal bloom effects on human health and
wider implications can be addressed using eDPSEEA as both a theoretical model and
stakeholder engagement process.
LINKING HUMAN HEALTH AND WELLBEING WITH WEATHER, CLIMATE
AND THE ENVIRONMENT WITH HARMFUL ALGAL BLOOMS (HABS)
Lora E Fleming1, Brian Golding2, Anthony Kessel4, Andy Haines3, Michael Depledge1, Anna
Cichowska4, Shakoor Hajat3, Christophe Sarran2, Nick Osborne1, Clive Sabel1,5, Trevor
Bailey5, Dan Bloomfield1,5
School, Truro, Cornwall, UK
2
UK Meteorological (Met) Office, Exeter, UK
3
London School of Hygiene and Tropical Medicine, London, UK
4
Public Health England, London, UK
5
University of Exeter, Exeter, UK
1
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
A large part of the global disease burden can be linked to environmental factors,
underpinned by unhealthy behaviours. However, research into these linkages suffers
from the lack of common tools and databases for carrying out investigations across
many different scientific disciplines to explore these complex associations. The MED MI
Partnership brings together leading organisations and researchers in climate, weather,
environment, and human health and wellbeing.
The main aim of the recently funded MED MI Project is to create a central data and
analysis source as an internet-based Platform which will be a vital new common
resource for medical and public health research in the UK and beyond. We will link
and analyse complex meteorological, environmental, and epidemiological data. This is
a vital step to translate this data and analysis resource into epidemiologic, clinical, and
commercial collaborative applications, and thus, improved human health and wellbeing
in a rapidly changing environment. Existing databases, currently stored in various
locations/organizations, will be combined enabling climate, weather and environment
data to be linked and analysed with human health and wellbeing data. With appropriate
confidentiality and ethical safeguards, the Platform will be available to researchers.
The “Climate, Coastal & Ocean Dynamics, and HABs: Blue Sky Demonstration Project”
will leverage new Met Office capabilities to link coastal and oceanographic processes
with remote sensing data to explore the possible link between HABs and climate change.
The aim will be to look at whether there is an association between recent climate
variability and the observed occurrence of blooms of the specific species sampled in
ongoing monitoring programmes. If associations are seen with this modelling, health
endpoint data from a variety of sources will be linked to explore acute and chronic
diseases possibly associated with HABs and residence relative to coasts and other
water bodies using time series and case-crossover analyses. The expansive scale in time
and space, as well as large sample sizes of the human health databases, will provide
sufficient data to explore the climate change-HAB-human health hypothesis and other
aspects (e.g. identification of potentially vulnerable populations, modelling for early HAB
warnings).
Page 142
DEVELOPMENT AND EVALUATION OF A DIRECT LC-MS/MS METHOD FOR
DETERMINATION OF DMSP IN BIOLOGICAL SAMPLES
Harold A. Flores Quintana1, Alison Robertson1 and Ronald P. Kiene2, 3
FDA, Gulf Coast Seafood Laboratory, Dauphin Island, AL 36528, USA
Department of Marine Sciences, LSCB-25, University of South Alabama, Mobile, AL
36688, USA
3
Dauphin Island Sea Lab, 101 Bienville Blvd, Dauphin Island, AL 36528, USA
1
2
Dimethylsulfoniopropionate (DMSP) is an organic sulfur zwitterionic compound
produced by a variety of marine micro- and macro-algae, and a few higher plants. This
compound is involved in oceanic carbon and sulfur cycling and is major precursor of
marine dimethylsulfide (DMS). DMSP has been found in a variety of harmful algae
including Alexandrium spp., Dinophysis spp., Prorocentrum spp., Gymnodinium spp.,
Karenia spp., Gambierdiscus spp., and may play an important role in toxin production.
Traditional quantification methods for DMSP are indirect and include base hydrolysis
and measurement of the cleavage product DMS, using gas chromatography (GC).
While sensitive, these methods may overestimate the DMSP concentration when other
dimethylsulfonium compounds are present.
This HILIC UPLC-MS/MS method is sensitive and provides a fast, direct, and specific
analysis of four dimethylsulfonium compounds. It allows simultaneous determination
of these compounds in a variety of biological matrices. Applications of this method may
improve our understanding of the complex role of DMSP in the oceanic sulfur cycling
and toxin production by harmful algal species.
Page 143
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ORAL PRESENTATIONS
POSTER
We developed a sensitive and specific ultra-performance liquid chromatography
tandem mass spectrometry (UPLC-MS/MS) method for direct measurement of
DMSP in a variety of biological matrices including seawater, phytoplankton cultures,
macroalgae, plants, and fish tissues. Hydrophilic interaction chromatography (HILIC)
was used to separate related dimethylsulfonium compounds including DMSP, 2-methylDMSP, dimethylsulfoxonium propionate (DMSOP), and gonyauline. Selected reaction
monitoring (SRM) in positive ion mode with multiple transition ions was employed
to unambiguously confirm and quantify DMSP and related compounds. The limit
of quantification for DMSP was 0.37 nM and with a linear range of 0.37 – 745 nM
(r2 > 0.998). Ionization suppression was observed in fish extracts but these matrix
effects were eliminated using standard dilution methods. DMSP extraction methods
were evaluated in fish tissue by spiking with 2-methyl DMSP as an internal standard.
Maximum recoveries were obtained with 100% methanol and yielded > 90% recovery.
Comparison of the UPLC-MS/MS method with the indirect GC method in phytoplankton,
macro algae, higher plants, and fish samples confirmed that the latter, overestimates
the DMSP concentrations in some instances. The highest levels of DMSP were found
in higher plants and phytoplankton; concentrations in fish were much lower, and were
species-specific.
A CHEMICAL ANALYSIS OF KAREnIA PAPILIOnACEA
Nicholas Fowler, Andrea Bourdelais, Carmelo Tomas, Elizabeth Elliott, Bob York, Daniel
Baden.
Center for Marine Science, UNCW, 5600 Marvin K. Moss Lane, Wilmington NC 28409
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Harmful algal blooms (HABs) occur worldwide and can produce devastating effects to
marine life, such as massive fish kills, death to marine mammals or contamination of
seafood for human consumption. One of the most widely studied organisms responsible
for HABs is the marine dinoflagellate Karenia brevis. This organism produces the highly
neurotoxic compounds known as brevetoxins. When ingested, these compounds
produce severe gastrointestinal maladies, as well as other neurological disorders such
as hot-cold sensorial reversal. A related dinoflagellate, Karenia papilionacea, is known
to co-bloom with Karenia brevis. However, K. papilionacea has received little attention
as a possible toxin producing species. Therefore, studies were undertaken, using a
combination of liquid-liquid extraction techniques along with HPLC, Mass Spectrometry,
NMR, and cytotoxicity assays to isolate compounds, test their toxicity and to perform
structural identification of the compounds produced by two isolated strains of K.
papilionacea
Results from these studies led to the isolation of a toxic extract from two isolated strains
of K. papilionacea (New Zealand and Delaware). Comparison studies of this isolate with
known compounds produced by K. brevis identified this compound as PbTx-2. These
results reveal, for the first time, the identity and quantities of a toxin produced by K.
papilionacea. This could provide an explanation for why the toxic bloom of 1993 in
New Zealand showed the presence of brevetoxins, without the presence of a known
brevetoxin producer. Full results of this research will be presented.
Page 144
TRANSLATIONAL RESPONSE TO HEAT STRESS IN THE FLORIDA RED
TIDE DINOFLAGELLATE, KAREnIA bREvIS
Kelly A. Fridey1,2, Jeanine S. Morey2, Matthew R. Paul3, Paul E. Anderson3, Rosemary
Jagus4, Allen R. Place4, and Frances M. Van Dolah2,1
Graduate Program in Marine Biology, College of Charleston, Charleston, SC 29412
3
Bioinformatics Research Group, Department of Computer Science, College of
Charleston, Charleston, SC 29424
4
Institute of Marine and Environmental Technology, University of Maryland, Baltimore,
MD 21202
1
2
Page 145
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ORAL PRESENTATIONS
POSTER
Karenia brevis is responsible for harmful algal blooms in the Gulf of Mexico that
cause extensive marine animal mortalities and human illnesses. K. brevis blooms are
particularly damaging when they persist at high density in coastal waters over long
periods of time. However, it is not yet known how K. brevis cells adapt to stresses
associated with changing coastal conditions. Previous work in our laboratory revealed
a lack of transcriptional activation of stress genes under conditions that induced stress
proteins. This is consistent with an emerging view that dinoflagellate gene expression
is regulated predominantly at the post-transcriptional level, in part by differential rates
of translation. The current project was undertaken to determine if stress responses are
mounted at the translational level. Translational activity can be inferred from polysome
profiles. Polysomes are mRNAs with multiple ribosomes attached and represent the
mRNA pool being actively translated, whereas ribosome-free mRNAs are translationally
inactive. In this study, triplicate cultures of K. brevis were exposed to a 5 °C heat shock
for a short time course (0, 30, or 60 min) to determine their translational response to
heat stress. Sucrose density gradient fractionation was used to separate polysomes
from ribosome-free RNA, detected spectrometrically by absorbance of 254 nm. The
abundance of polysomes decreased rapidly in response to heat shock, indicating a
reduction of translation, with the lowest polysome abundance found at 60 min of
exposure. RNA was isolated from the translationally active fractions at each time point
and RNA-seq analysis was performed on an Illumina Hi-Seq2000 sequencer at a depth
of 15 million reads per sample. A reference K. brevis transcriptome was assembled from
113 million Illumina reads (50 bp, paired ends) using Trinity. This assembly contains
127K unique contigs. Quantitative read mapping to the reference transcriptome,
currently in progress, will allow the assessment of whether stress response gene
transcripts are specifically recruited to the actively translated RNA pool following heat
shock.
STRUCTURE ACTIVITY RELATIONSHIP OF BREVENAL DERIVATIVES
Allan Goodman, Jennifer McCall, Karl Jacocks, Alysha Thompson, Daniel Baden and
Andrea Bourdelais
Center for Marine Science, UNCW, 5600 Marvin K. Moss Lane, Wilmington, NC 28409
ORAL
PRESENTATIONS
POSTER
PRESENTATIONS
The marine dinoflagellate Karenia brevis is associated with the phenomenon known as
Florida Red Tide and produces a variety of ladder frame polyether compounds, including
the highly active neurotoxic brevetoxins. In addition to these toxins, K. brevis produces
brevenal, a compound shown to possess antagonistic activity to the brevetoxins. In
asthmatic sheep models, brevenal was able to both inhibit and reverse brevetoxin
induced bronchoconstriction. Furthermore, when administered alone, brevenal was
able to increase tracheal mucosal velocity. These findings have led to the development
of brevenal as a potential treatment for cystic fibrosis and the compound is in late stage
preclinical studies.
To further understand the mechanism of action of brevenal and potentially develop
a second generation drug candidate, a series of brevenal derivatives were prepared
through modification of the aldehyde moiety. These derivatives include aliphatic,
aromatic and heteroaromatic derivatives. The brevenal derivatives were then tested in
in vitro binding assays to determine the ability of the compounds to displace brevetoxins
and brevenal from their native receptors. From the data generated, it was found that
a large variety of functionalities were tolerated, but binding at the brevenal receptor
remained mostly unchanged. However, compounds containing a meta-substituted
phenylhydrazide led to ligands with the greatest ability to displace brevetoxin. For
instance, in the competitive binding assay for brevetoxin, the 3-chloro- (1) and
3-methoxyphenylhydrazides (2) displaced the brevetoxin with 7 and 8 nM affinities
respectively, compared to 1133 nM for brevenal in the same assay. Additionally, a biotin
derivative was prepared to aid in a streptavidin mediated isolation of the brevenal
receptor. Full findings from these studies will be presented.
Page 146
DETERMINING FUNCTIONAL EFFECTS OF BREVETOxIN AND
BREVETOxIN ANTAGONISTS
Meghan Grandal1, Alison Taylor1, Daniel G. Baden2 and Andrea Bourdelais2
Department of Biology and Marine Biology, University of North Carolina at Wilmington,
Wilmington NC 28409 USA
2
Center for Marine Science, University of North Carolina at Wilmington, Wilmington NC
28409 USA
1
Page 147
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October 2013
POSTER
ORAL PRESENTATIONS
Brevetoxin (PbTx) is lipophilic polyether compound produced by the dinoflagellate
Karenia brevis that is responsible for massive fish kills in the Gulf of Mexico and can
result in neurotoxic shellfish poisoning in humans after consumption of shellfish
contaminated with the toxins. The physiological effects of PbTx are due to its activity
on neuronal sodium (Na+) channels where it binds selectively to site 5 of voltage gated
sodium channels. Binding of brevetoxin to this receptor results in hyperexcitability by
increasing the mean open time of the channel and inhibition of channel inactivation,
resulting in a range of neuroexcitatory symptoms. The nontoxic brevetoxin derivative,
brevenal, inhibits the icthyotoxic and cytotoxic effects of PbTx by competitively binding
to Na+ channels. Attention has been drawn to brevenal as a potential therapy for specific
channelopathies due to its affinity for Na+ channels and nontoxic nature. Specifically,
brevenal has been shown to reduce bronchoconstriction and increase mucus clearance
in an asthmatic sheep model more effectively than current marketed drugs for cystic
fibrosis (CF). A number of other naturally and synthetically produced PbTx derivatives
are currently being tested for pharmaceutical potential. In this study, cell assays have
been developed to measure functional responses at the cellular level, using quantitative
methods. Amperometry and mass spectrometry were used to quantify catecholamine
release from PC12 cells; both brevetoxin and brevenal stimulate secretion of the
neurotransmitter dopamine. This suggests that either both compounds affect Na+
dynamics and membrane potential upon binding to their receptor sites, or that entrance
of the toxin through cell membrane (as shown by studies with fluorescently labeled
PbTx derivatives) alters intracellular Ca2+ dynamics and cellular pathways leading to
neurotransmitter secretion. Ion-sensitive dyes are currently being used to measure
change in intracellular Na+ and Ca2+ after PbTx and brevenal treatment to characterize
downstream signaling cascades and the mechanism by which these compounds
stimulate secretion. The ability to stimulate secretion makes PbTx derivatives a
compound of interest, not only for CF, but for a range of other neuro- and endocrine
degenerative diseases.
ELUCIDATING THE GENETIC DIVERSITY OF mICROCySTIS AERuGInOSA
WITHIN A TOxIC BLOOM IN SOUTH CAROLINA COASTAL PONDS USING
454-PYROSEQUENCING TECHNOLOGY
Dianne I. Greenfield 1,2,3; William J. Jones 2,4; Sarah Hogan 3; Chuck Keppler 3
Belle W. Baruch Institute for Marine and Coastal Sciences, University of South Carolina,
Columbia SC 29208, USA
2
Marine Sciences Program, University of South Carolina; Columbia SC 29208, US3Marine
Resources Research Institute, Charleston SC 29412, USA
4
Arnold School of Public Health, University of South Carolina, Columbia SC 29208, USA
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
Coastal South Carolina (SC) is currently undergoing some of most rapid development
in the United States, including residential neighborhoods and recreational (golf course)
construction with detention ponds as catchments for stormwater runoff. Due to
their poor flushing, SC detention ponds accumulate nutrients and fertilizers, making
them susceptible to eutrophication and harmful algal blooms (HABs). Cyanobacteria
HABs are particularly common, with Microcystis aeruginosa being among the most
abundant species. A particularly dense (> 6 x 105 cells/ml) and toxic multi-specific
cyanobacteria bloom occurred during August-September of 2011 in one such pond,
with M. aeruginosa as a dominant component. We evaluated the spatial and genetic
diversity of a key gene critical for the production of the hepatoxin microcystin (mcyD)
within this bloom. Relevant enviromental parameters (standard water quality, nitrogen,
phosphorus, chlorophyll a, microcystin concentrations, etc.) were also evaluated from
multiple sampling stations. To assess spatial variability within the bloom, each sample
was individually barcorded using a unique multiplex identifier (MID) sequence, and DNA
was amplified using primers specific to mcyD. Products were subsequently sequenced
using Roche/454 pyrosequencing technology. Initial results revealed multiple and
distinct clades, suggesting a high degree of intraspecific variability. Here we discuss our
findings, the various challenges encountered while applying this sequencing approach
to HAB research, and the environmental factors that may mediate cyanobacteria bloom
development.
Page 148
SUCESSFUL OCEAN OBSERVATORIES AT WORK ON A HARMFUL ALGAL
BLOOM ON THE WEST FLORIDA SHELF
Alan Hails1, Gary Kirkpatrick1, Chad Lembke2, Robert Currier1, L. Kellie Dixon1
University of South Florida, 140 7th Avenue South, St. Petersburg, FL 33701, USA
1
2
In October, 2012, during an evolving Red Tide bloom event, Mote Marine Laboratory
(Mote) and University of Southwest Florida (USF) deployed multiple instruments and
ocean platforms to gather data needed to characterize and study it. The autonomous
underwater vehicle (AUV) portion of the resources for this collaborative project was
sponsored by the Gulf of Mexico Coastal Ocean Observing System (GCOOS) and State of
Florida Fish and Wildlife Conservation Commission (FWC). The GCOOS portion of the
project had the additional objective of demonstrating real-time telemetering of multiple
AUV and sensor data to the GCOOS data servers where visualizations of AUV data fused
with satellite imagery and ocean models could be produced.
The Mote AUV was equipped with a flow-thru
CTD and with the Mote-developed Optical
Phytoplankton Discriminator. The OPD is a
mobile water sampling spectrophotometer
employed to quantify the many species of
the Gulf phytoplankton community, including
red tide. The USF AUV was equipped with
a similar CTD as well as with Fluorometric
sensors for chlorophyll, CDOM, and backscatter
and with oxygen and irradiance sensors. The
surface vessels took CTD measurements and
obtained water samples returned to the lab
for HPLC analysis of toxins and pigments and
for direct cells/liter measurements of red tide.
Ashore, remote imagery data and ocean model
predictions were produced to guide the data
collection efforts.
We describe the many resources employed during this event, the numerous types of
data collected and how they were combined and compared to learn about the origins,
3-D location and progression and strength of the bloom, and how the data was ported to
the Gulf regional observatory for presentation and sharing in real-time.
Page 149
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ORAL PRESENTATIONS
POSTER
During the period from October 6 through October 23, 2012 the AUV Waldo operated
by Mote and the AUV Bass, operated by USF, were concurrently “flown” through an
area 15 nm offshore of Englewood, Florida to 50 nm offshore of Naples, Florida. Within
that area a concentrated data collection effort took place 5-11 nm SW of Sanibel Island.
Also concurrently, under other programs sponsored by FWC, manned surface research
vessels were deployed by Mote on October 4, 9, and 10.
FACILITATING IMPLEMENTATION OF THE RECEPTOR BINDING ASSAY
FOR PSP
Sherwood Hall1, Frances Van Dolah2, Gregg Langlois3, and Leanne J. Flewelling4
U. S. Food and Drug Administration, 1US FDA Center for Food Safety and Applied
Nutrition, 5100 Paint Branch Parkway, College Park, MD 20740, USA.
2
3
California Department of Public Health, Environmental Management Branch, 850
Marina Bay Parkway, Richmond, CA 94804
4
100 Eight Avenue SE, St. Petersburg, FL 33701, US
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
The receptor binding assay (RBA) for Paralytic Shellfish Poison (PSP) toxins offers
excellent sensitivity, high throughput, and a reliable measure of potency. Several steps
have been taken recently to facilitate its implementation. The RBA is now an AOAC
International-validated method, OMA 2011.27. The radiolabeled saxitoxin required
for the RBA is now commercially available. An exemption has been obtained from the
Nuclear Regulatory Commission (NRC) so that labs wishing to use the RBA can now
purchase, use, and dispose of the radiolabeled saxitoxin without an NRC license. The
membrane suspension needed for the method is now also commercially available,
frozen in vials, ready for use in the assay. Finally, it has been found that the multiwell
plate counters needed for the assay are available used/reconditioned for about a third of
the cost of a new counter, significantly lowering the cost of implementing the RBA.
Page 150
AN ExPERIMENTAL ASSESSMENT OF THE IMPORTANCE OF PROPAGULE
PRESSURE AND COMMUNITY RESISTANCE ON THE INVASION SUCCESS
OF THE TOxIGENIC HAPTOPHYTE PRymnESIum PARvum
Richard M. Zamor and K. David Hambright
Plankton Ecology and Limnology Laboratory and Program in Ecology and Evolutionary
Biology, Department of Biology, University of Oklahoma, Norman, OK
In recent years, harmful algae have bloomed in habitats where they were previously
unknown to exist, suggesting that they may be invasive species. Alternatively, harmful
algae may have always been present in these systems, perhaps as members of the
microbial rare biosphere, and these systems have changed environmentally (e.g., via
nutrient pollution, water resource overexploitation, or climate change) to become
more amenable to population explosions by these previously undetected species.
The toxigenic and ecosystem disruptive algal bloom species, Prymnesium parvum, is
one such species. Originally described in marine coastal environments, P. parvum has
successfully established populations in inland brackish systems globally, causing major
ecological devastation, exemplified by massive fish kills. Thus, P. parvum represents
a model organism for studying potential invasion and range expansion in a microbial
species. If P. parvum and other harmful algae can in fact be invasive, then we would
expect that general ecological understanding and theory derived from the study of
multi-cellular invaders should be applicable to microbes as well.
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October 2013
ORAL PRESENTA
PRESENTATIONS
POSTER
Two major factors implicated in the establishment success of invasive species are
propagule pressure and community resistance to invasion. The former represents
a game of numbers, with generally larger invading populations or populations with
repetitive invasions having greater chances for successfully establishing reproducing
populations in new habitats, while the latter is embodied in Hutchinson’s n-dimensional
niche theory, where the number of niches available for invasion is dependent on the
diversity of the community being invaded. In experimental mesocosms, we manipulated
community diversity in a natural phytoplankton assemblage (isolated from an inland
lake subject to frequent P. parvum blooms, and determined by high-throughput
pyrosequencing) by altering resource availability. We then subjected the manipulated
communities, in which P. parvum was not at detectable levels, to different levels of
propagule pressure by P. parvum. Our results indicate that, in the environmentally
suitable habitats of our experimental systems, neither community diversity nor
resource availability had any effect on invasion success by P. parvum. However, our
results documented a significant role for propagule pressure in the establishment of P.
parvum populations in these experimental systems. Although it is difficult to extrapolate
laboratory results to natural systems directly, our results indicate that general ecological
principles thus far developed from the study of macrobial invasive species can provide a
solid framework for better understanding what appear to be major range expansions in
P. parvum and other harmful algae.
SEASONAL PATTERNS IN NUTRIENTS AND PHYTOPLANKTON IN
QUARTERMASTER HARBOR, AN ENCLOSED PUGET SOUND EMBAYMENT
WITH RECURRENT HARMFUL ALGAL BLOOMS.
Gabriela Hannach1, Kimberle Stark2 and Cheryl Greengrove3
Environmental Lab, King County Department of Natural Resources and Parks Seattle,
WA, USA
2
King County Department of Natural Resources and Parks, Seattle, WA, USA
3
Department of Environmental Science, University of Washington, Tacoma, WA, USA
1
POSTER
PRESENTATIONS
ORAL PRESENTATIONS
Quartermaster Harbor, an enclosed bay within the Puget Sound Central Basin, has
poor tidal flushing and historically low levels of dissolved oxygen, leading King County
to initiate a nitrogen management study with the goal to identify key factors affecting
nitrogen loading in the bay. Additionally, recurrent blooms of the dinoflagellate
Alexandrium catenella occurring in spring and fall have been shown to coincide with
elevated levels of saxitoxin in shellfish and the frequent closure of shellfish beds to
harvesting. Bottom surveys have revealed unusually high concentrations of A. catenella
cysts in Quartermaster Harbor surface sediments, providing a ready source of motile
cells under favorable conditions. Blooms of the toxigenic diatom Pseudo-nitzschia
are also common in the bay. These combined findings have prompted a number of
investigations aimed at improving our understanding of the relationship between the
distinct environmental conditions present at Quartermaster Harbor and the seasonal
dynamics of its phytoplankton populations.
Sampling of the water column at Quartermaster Harbor is ongoing and consists primarily
of a combination of a) routine monthly or study-specific water samples for water quality
parameters, chlorophyll-a fluorescence and nutrients, and b) moorings that provide
continuous measurement of water quality parameters and chlorophyll-a fluorescence.
Routine phytoplankton analysis is conducted bi-weekly from March to November.
A comparison with open water sites indicated that the bay is characterized by an
extended bloom season, typically spanning from March or earlier and into October.
The shallow inner harbor waters warm up earlier in the year and stay warmer longer,
contributing to the extended bloom season. A sharp reduction in surface water
nutrients follows the early spring blooms, and inorganic nitrogen tends to remain
depleted throughout the summer. Large diatom blooms have also led to silica depletion.
As is characteristic of many estuarine systems, flagellate populations increase in
late summer/early fall in response to limited inorganic nutrients and warmer water
temperatures. In contrast to other sites, where Alexandrium catenella rarely forms
blooms, in Quartermaster Harbor it is often one of the dominant species during this
time. Results from these water property and phytoplankton community studies with a
focus on the occurrence of Alexandrium catenella and timing of shellfish bed closures
will be presented.
Page 152
LINKING FLORIDA RED TIDES TO HUMAN HEALTH EFFECTS: DATA
ISSUES AND PRELIMINARY SIGNALS
Porter Hoagland1, Di Jin1, Andy Beet1, Andy Reich2, Barbara Kirkpatrick3,4, Steve Ullmann6,
Sharon Watkins2, Lora E Fleming4,5,7
Marine Policy Center, Woods Hole Oceanographic Institution, Woods Hole, MA 02543,
USA
2
Aquatic Toxins Program, Environmental Health, Florida Department of Health,
Tallahassee, FL 32399, USA
3
4
Department of Epidemiology and Public Health, Miller School of Medicine, University of
Miami, Miami, FL 33177 USA
5
Sciences, University of Miami, Miami, FL 32399, USA
6
Miami, FL 33177 USA
7
European Centre for Environment and Human Health, Peninsula College of Medicine
and Dentistry, Truro, Cornwall, UK
1
Page 153
S
October 2013
ORAL PRESENTATIONS
POSTER
Florida red tides due to harmful blooms of the dinoflagellate Karenia brevis are known
to cause respiratory and gastro-intestinal illnesses in humans and marine mammals. In
humans, these illnesses can lead to a range of health impacts as reflected in physician
appointments, emergency department (ED) visits, and hospital admissions. Public
health specialists would like to understand further the nature of the human health
risks of Florida red tide across illness severities, but to date there is only a limited body
of research on this topic. Kirkpatrick et al. (2006, 2009) found that respiratory and
gastrointestinal ED visits in a Florida coastal community are associated with Florida red
tides in Sarasota County. Hoagland et al. (2009) used an exposure-response framework
to estimate the effects of Florida red tide on weekly ED visits at this hospital for
respiratory illnesses during 2001-2006. These authors found that a local measure of in
situ K. brevis cell counts lagged by one week could explain ED visits when controlling
for temperature, influenza cases, pollen, and a measure of tourist visits. In our current
study of the dynamics of HABS and human communities, we examine the human health
risks of Florida red tide for both respiratory and gastrointestinal illnesses for both ED
visits and hospitalizations over a longer period of time, across a wider geographic area
of Florida, and stratified by age. Because of limitations in the geographic monitoring for
K. brevis blooms, we have compiled panel data on the location and duration of closures
due to Florida red tide for 43 shellfish harvesting areas (SHAs) along the Florida Gulf
Coast. We discuss some of the difficulties in compiling data on exposures and responses,
and we report on the preliminary results of the analysis. When controlling for resident
population size, tourism, and seasonal effects, we find that ED visits for respiratory
illnesses can be explained by the number of monthly closures in coastal counties of
proximate SHAs as a measure of the occurrences of Florida red tide blooms. These
findings are geographically restricted to four mid-coast counties (Pinellas, Hillsborough,
Sarasota, and Lee), and further limited to persons ≥ 55 years. A relationship between the
red tide measure and gastro-intestinal illnesses may exist, but is more ambiguous.
DEVELOPMENT OF AN LCMS ASSAY FOR DETERMINATION OF AMINO
ACID CONCENTRATION IN KAREnIA bREvIS
Tanya Hogue, Nicholas Fowler, Daniel Baden and Andrea Bourdelais
Center for Marine Science, University of North Carolina at Wilmington, 5600 Marvin K.
Moss Lane, Wilmington, NC 28409 USA
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Karenia brevis is a marine dinoflagellate responsible for the production of brevetoxins
and other ladder frame polyether (LFP) compounds. The brevetoxins are a class of
neurotoxins and are associated with massive fish kills, marine mammal mortality and
adversely affect tourism and seafood production, especially in the Florida coastal
waters in the Gulf of Mexico. Previous research has shown that K. brevis exposed to
sudden reduction in salinity levels, such as may be experienced when blooms approach
estuarine environments, leads to a rapid increase in LFP production. Therefore, it has
been hypothesized that the LFPs are utilized by K. brevis in an osmoregulatory fashion.
An alternate hypothesis is that brevetoxins are not involved in acclimation to salinity
changes, rather compounds such as: lipids, dimethylsulfoniopropionate, ions and amino
acids may be the osmoregulators in phytoplankton
Therefore, studies were initiated to develop an LCMS based assay to determine
intracellular concentrations of amino acids produced by K. brevis under salinity stress.
One of the key challenges of this research has been development of a suitable method
for isolating and detecting the very low concentrations of free amino acids found inside
K. brevis cells. In particular, extraction and quantification of amino acids from K. brevis
has required overcoming various obstacles such as: competition of intracellular amino
acids and salts found in sea water with ion exchange columns, difficulty in separation
of amino acids on traditional HPLC columns due to their polar properties, instability
of amino acid conjugates and the inability to use non-volatile buffers with the LC-MS
system available at UNCW. Various methods for isolation of amino acids from sea
water, conjugation techniques to make the amino acids less polar and LC conditions
were investigated. Development of methods for isolation and detection of amino acids
produced by K. brevis will be presented.
Page 154
COMPETITIVE AND MUTUALISTIC INTERACTIONS BETWEEN TOxIC
AND NON-TOxIC STRAINS OF THE HARMFUL ALGAL BLOOM-FORMING
mICROCySTIS AERuGInOSA
Sarah M. Holmes1, Mark W. Silby1, and Pia H. Moisander1
University of Massachusetts Dartmouth, Department of Biology, 285 Old Westport Rd.,
N. Dartmouth, MA 02747, USA
1
Page 155
S
October 2013
ORAL PRESENTATIONS
POSTER
Microcystis aeruginosa is a common bloom-forming cyanobacterium that is found
in eutrophic bodies of fresh or brackish water throughout the world. Both toxic and
nontoxic strains can be found within the same bloom with composition changing
over the course of the bloom. Some blooms are considered to be HABs as the toxic
strain produces a hepatotoxin, microcystin, which can be lethal to livestock that
accidentally drink contaminated water and has non-lethal effects on humans including
gastrointestinal discomfort and contact dermatitis. The genetic mechanism by which
some Microcystis strains are toxic is known but the ecophysiological reasons are not
fully understood. The fact that toxic and nontoxic strains appear together and their
ratios predictably change over time suggests the possibility of a specific interaction
between the strains which may involve signaling by microcystin. In this ongoing
study we are investigating the competitive and mutualistic interactions between the
toxic and nontoxic Microcystis strains. We propose that the toxic strain is providing
a benefit due to the toxin while the nontoxic strain saves energy by not participating
in toxin production whereby that energy is directed toward reproduction while still
gaining the benefit of the toxin. We are investigating variation in growth rate between
mono- and co-cultures of the toxic PCC7806 strain and nontoxic KLA2 strain under
varying light conditions in order to identify if the toxin increased fitness under high
light. Initial experiments indicate that the co-culture has an increased growth rate in
comparison to the mono-cultures which lends preliminary support to the concept of
the increased benefit of the toxic strain to the nontoxic strain. Novel qPCR primerprobe sets were designed to target the two strains used in the co-culture experiments,
using the published genome for PCC7806 and the draft genome for the non-toxic strain
from our laboratory. Initial qPCR runs with the primer sets indicate that they will be
useful in determining the abundance of toxic and nontoxic cells grown together since
these strains are visually indistinguishable. In addition to growth rate data, we plan
to look at the transcriptomes of mono- and co-cultures with the prediction that there
will be differential transcription between these cultures. The results will improve our
understanding of the role of the toxin while investigating communication between
strains and influence of variations of their relationship under different environmental
conditions. This knowledge will be useful in monitoring and managing cyanoHABs and
protecting water resources.
GEITLERInEmA SP. PRODUCES A NOVEL ICHTHYOTOxIC
CYANOBACTERIAL TOxIN
I-Shuo Huang1, Peter Moeller2, Dale Casamatta3, Danielle Gutierrez1, and Paul V. Zimba1
Texas A&M University—Corpus Christi, Center for Coastal Studies, 6300 Ocean Dr,
Corpus Christi, TX 78412
1
NOAA, Hollings Marine Laboratory, 331 Ft. Johnson Rd, Charleston, SC 29412
2
University of North Florida, 1 UNF Dr, Jacksonville, FL 32224
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
3
Cyanobacterial toxins are best known in freshwater systems, with only two toxins
known from marine systems. It is likely that many other marine cyanobacterial toxins
are undescribed. A marine Geitlerinema sp. was isolated from an aquatic animal
rearing facility having mysid mortality events. The cyanobacteria possibly originated
from Chesapeake Bay, MD, or Corpus Christi Bay, TX. Unialgal bulk cultures were used
to produce sufficient material to isolate the toxin. Isolation of the unknown toxin was
accomplished by bioassay-guided fractionation using HPLC-MS. The morphology of the
Geitlerinema isolate does not match other known marine taxa. Molecular phylogenetic
analysis in addition to morphology will be used for species identification. The toxin has
a unique mass-to-charge ratio of 475.3 amu that does not match any known toxins.
Structural confirmation by TOF-MS and NMR is ongoing. Toxicity of fish, mysids, and
copepods occur after toxin exposure; LD50 for copepods was 158 mg/L. The LD50 values
are helpful for understanding the species response once exposed to the toxin and the
relative potency of the toxin compared to other toxins.
Page 156
BIOASSAY-GUIDED PURIFICATION OF A BIOACTIVE CAROTENOID
FROM THE FRESHWATER CYANOBACTERIUM, CyLIndROSPERmOPSIS
RACIbORSKII
Asha Jaja-Chimedza1, Miroslav Gantar2, and John P. Berry1
Department of Chemistry and Biochemistry, Florida International University, 3000 NE
151st Street, North Miami, FL 33181 USA
2
Department of Biological Sciences, Florida International University, 11200 SW 8th Street,
Miami, FL 33199, USA
1
Page 157
S
October 2013
ORAL PRESENTATIONS
POSTER
Cyanobacteria (“blue-green algae”) are recognized to produce a wide array of bioactive
compounds. Cylindrospermopsis is a known toxigenic genus of freshwater cyanobacteria
that produces a number of toxic metabolites, particularly including the water-soluble
toxins cylindrospermopsin and saxitoxin, and is frequently associated with toxic
cyanobacterial blooms. Lipophilic extracts from a strain of C. raciborskii that does
not contain these known toxins, was previously shown to inhibit the development of
zebrafish (Danio rerio) embryos as a relevant freshwater teleost fish species. Bioassayguided fractionation using the zebrafish embryo, as a model of so-called “developmental
toxicity,” was subsequently utilized in the purification of the apparent toxic metabolite.
Using this approach, an apparent pigment was identified as the bioactive compound
and partial chemical characterization suggested that this compound was a carotenoid.
Carotenoids are widely distributed among cyanobacteria, and generally associated with
harvesting light and protection against photooxidative damage, but they have not been
previously associated with any bioactivity. Our studies have shown that this compound
clearly inhibits vertebrate development in zebrafish embryos, and thus may, indeed,
contribute to the toxicity of this species. Chemical structure and associated toxicology
of this presumed carotenoid will be presented. Due to the frequent occurrence of C.
raciborskii in association with cyanobacterial harmful algal blooms, these bioactive
compounds may pose a previously unrecognized environmental and health risk.
INVOLVEMENT OF MULTIPLE EIF4ES IN MRNA RECRUITMENT IN
DINOFLAGELLATES
Grant D. Jones1, 2, Tsvetan R. Bachvaroff2, Allen R. Place2 and Rosemary Jagus2
University of Maryland, Baltimore, 620 W. Lexington St., Baltimore, MD 21201, USA
1
Institute of Marine and Environmental Technology, University of Maryland Center for
Environmental Science, 701 E. Pratt St., Baltimore, MD 21202, USA
2
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Insight into the molecular mechanisms that control the growth and persistence
of dinoflagellate blooms is critical for the development of mitigation strategies.
Organization of the genome and gene expression in dinoflagellates has been shown
to be distinct from other eukaryotes. A wide range of studies has implicated mRNA
recruitment as a major site of regulating gene expression in dinoflagellates. However,
relatively little is understood regarding this process in these organisms. It is known
that a short 22-nucleotide Spliced Leader exon is spliced from a capped small nuclear
RNA and transferred to pre-mRNA, thereby providing the 5’-terminal end and the cap
structure to mature mRNAs. Given this modification of mRNAs and a non-traditional cap
structure, we have focused our investigation on the cap-binding translational initiation
factor eIF4E.
Similar to plants and metazoans, dinoflagellates express a variety of eIF4E family
members that have the potential to function in the recruitment of mRNA to the
ribosome or in mRNA regulation. Our phylogenetic analysis of RNAseq data on multiple
dinoflagellate species found three separate clades of eIF4E. Clade 1 eIF4Es displayed
the highest expression and contained up to seven conserved subtypes. Clades 2 and
3 had lower expression and each contained two or three more divergent subtypes.
To compare the functions of eIF4Es from each clade, we synthesized four different
members of Clade 1, one member of Clade 2, and one member of Clade 3 from
Karlodinium veneficum, and we cloned them into a mammalian expression vector for
in vitro translation in rabbit reticulocyte lysate. Using binding to methyl7GTP-agarose
beads as a measure of affinity to the 5’-mRNA cap, we demonstrated differential binding
of the eIF4Es from different clades.
Clade 1 eIF4Es bound to methyl7GTP beads. In contrast, members of Clades 2 and 3 did
not bind to the methyl7GTP beads. This is consistent with Clade 1 eIF4Es representing
the functional initiation factors. Clade 1 eIF4Es contain extended amino acid stretches
between the structural units of the eIF4E core. Interestingly, these regions show
marked heterogeneity between the Clade 1 subtypes. From this, we may anticipate
differing functions between the subtypes, such as selectivity for specific mRNAs.
Cap binding characteristics of eIF4Es from Clades 2 and 3 suggest they may fulfill
regulatory functions. This work is establishing additional tools for investigating the posttranscriptional regulatory mechanisms in dinoflagellates.
Page 158
EFFECTS OF NUTRIENTS AND GRAZING MORTALITY ON THE
ABUNDANCE OF AuREOumbRA LAGunEnSIS DURING A FLORIDA
BROWN TIDE BLOOM IN 2012
Yoonja Kang, Florian Koch, and Christopher J. Gobler
School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook, NY
11794-5000, USA
Page 159
S
October 2013
ORAL PRESENTATIONS
POSTER
During the summer of 2012, a brown tide bloom caused by Aureoumbra lagunensis
emerged in the Mosquito and Indian River Lagoons, Florida, USA. To assess contribution
of top-down (e.g. grazing) and bottom-up (e.g. nutrients) controls in shaping
phytoplankton community composition during the bloom, nutrient amendment and
dilution-style zooplankton grazing experiments were conducted during this brown tide
bloom in the southern Mosquito Lagoon. During the study, A. lagunensis represented
from up to 90% of total phytoplankton biomass and cell densities exceeded 2 x 106 cells.
The brown tide bloom was associated with high levels of dissolved organic nitrogen
(DON, > 80 μM) and low levels of dissolved inorganic nitrogen (DIN, < 2 μM). The low
levels of DIN and low DIN:DIP ratio were indicative of a inorganic nitrogen-limited
system, while the high DON:DOP ratio indicated there was an abundant supply of
organic nitrogen. Nutrient amendment experiments demonstrated that the addition
of DIN and DIP favored the growth of non-brown tide eukaryotes and phycocyanin
and phycoerythrin cyanobacteria population, whereas the growth of A. lagunensis
was not significantly affected by the addition of the same nutrients. These results
suggested that ambient nutrients were ideal for the growth of A. lagunensis during
the blooms. Dilution experiments demonstrated that microzooplankton grazing rates
on A. lagunensis were significantly lower than those on other major phytoplankton
populations. Low grazing pressure likely allowed A. lagunensis to out-compete other
phytoplankton populations. Given these findings, it is concluded that both top-down
(selective microzooplankton grazing) and bottom-up (high DON and low DIN system)
promoted A. lagunensis brown tides in the southern Mosquito Lagoon, FL, in 2012.
EFFECTS OF CHRONIC LOW-DOSE DOMOIC ACID ExPOSURE ON
MITOCHONDRIAL FUNCTION IN MICE
Preston Kendrick1, Kathi Lefebvre2, David Marcinek1
Department of Radiology, University of Washington, Seattle WA 98109
Environmental and Fisheries Science Division, NOAA NMFS, Northwest Fisheries
Science Center, Seattle, WA 98112
1
POSTER
PRESENTATIONS
ORAL PRESENTATIONS
2
The neurobehavioral excitotoxic effects of acute exposure to domoic acid have been
well documented in both field (marine mammal) and lab (rat, mouse) studies, but the
impacts of long-term, sub-clinical exposure to domoic acid have not been addressed
in a mammalian system. In zebrafish, repeated sub-clinical doses of domoic acid have
been shown to impact mitochondrial function. These impacts included increased
mitochondrial content in exposed fish, but decreased O2 flux per unit mitochondria,
indicating that the mitochondria functioned less efficiently in exposed fish. Chronic subclinical exposure to domoic acid also induced an immune reaction in zebrafish, eliciting
the production of a domoic acid specific antibody detectable in serum. Similar impacts
from chronic exposure are expected in a mammalian system. To elucidate these
responses, mice were exposed to sub-clinical doses of domoic acid once a week for six to
twelve weeks. At the end of six weeks, samples of cerebellum were taken for testing of
mitochondrial function and content. Additionally blood was collected after twelve weeks
of exposure to test for the presence of a domoic acid specific antibody. Preliminary
results indicate a decrease in mitochondrial function, however the presence of a domoic
acid specific antibody was not detected.
Page 160
CHANS: PUBLIC PERCEPTION OF FERTILIZER ORDINANCES AND
FLORIDA RED TIDES
Karen Scheller1,2, Barbara Kirkpatrick2,3,4, Kate Kohler2, Lora E Fleming3,4,5, Margaret
Byrne3, Jamie Studts6, Andy Reich7, Gary Kirkpatrick2, Steve Ullman8, Gary Hitchcock4,
Porter Hoagland9.
University of Notre Dame, Notre Dame, Indiana
3
Dept of Epidemiology, Miller School of Medicine, University of Miami, Miami, FL
4
Oceans and Human Health Center, RSMAS, University of Miami, Miami, FL
5
European Centre for Environment and Human Health, University Exeter Medical School,
Truro, Cornwall, UK
6
Department of Behavioral Science, University of Kentucky, Lexington, KY
7
Aquatic Toxins Program, Environmental Health, FL Dept of Health, Tallahassee, FL
8
Business School, University of Miami, Miami, FL
9
Marine Policy Center, Woods Hole Oceanographic Institute (WHOI), Woods Hole, MA
1
2
Methods: In 2012, reported knowledge and behaviors around HABs and nutrients were
evaluated in 305 permanent and seasonal (snowbirds) residents in a coastal community
regularly affected by blooms of Florida red tide and with a municipal fertilizer ordinance.
Results: The overall study population was predominantly older with a mean age (±SD)
of 56 ± 14.2 years (range: 19-86 years), female (57.9%), and highly educated. Only 34%
of the study population had any knowledge of the fertilizer ordinance, yet 45.9% then
stated that the purpose of the ordinance was to reduce red tide. 20% of participants
reported applying fertilizer to their lawns themselves and another 25% reported that
their lawn company applies fertilizer. There were no significant differences between the
residents and snowbirds regarding their knowledge and behavior with regards to HABs,
fertilizer use, or knowledge of policy.
Conclusions: This study demonstrates the gaps between public perceptions, and the
implementation of public policy. With poor awareness of the fertilizer ordinance,
the effectiveness of the ordinance is unknown. Given a scenario of unproven and
controversial science, we recommend that local communities consider surveying
stakeholder use and knowledge of potential impacts from fertilizer application prior to
policy implementation.
Page 161
SS
October 2013
ORAL PRESENTATIONS
POSTER
PRESENTATIONS
Background: Karenia brevis (Florida red tide), the most common marine HAB, produces
natural toxins that causes morbidity and mortality in exposed animals and humans.
A possible but controversial cause of Florida red tide blooms is coastal inputs of
macronutrients, nitrate and phosphate, through runoff of fertilizers. Despite scientific
uncertainty about causality, in 2007 some coastal communities instituted local
nutrient-restriction policies (including a municipal ordinance regulating lawn fertilizer
applications) in response to repeated and severe Florida red tide events.
WHERE AND HOW DO INDIVIDUALS RECEIVE AND PREFER TO RECEIVE
INFORMATION ON FLORIDA RED TIDE?
Kate Kohler1, Margaret M Byrne2, Jamie Studts4, Ben Whitenack1,5, and Barbara
Kirkpatrick1,2,3
Dept of Public Health Studies, Miller School of Medicine, Univ of Miami, Miami, FL
3
Rosenstiel School of Marine and Atmospheric Sciences, Univ of Miami, Miami, FL
4
Department of Behavioral Science, Univ of Kentucky, Lexington, KY
5
University of Pennsylvania, Philadelphia, PA
1
2
Background: Harmful algal blooms (HABs) pose challenging issues for public health
communication efforts. Many products such as brochures, pamphlets, websites and
Facebook pages have been created over the years to dissemination information
to interested parties. This is certainly true with Florida red tide, ie Karenia brevis,
blooms. However, to our knowledge, those potentially affected by HABs have not been
surveyed for their preferences regarding information delivery mechanisms and trusted
information sources when seeking information regarding HABs.
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Methods: We surveyed 185 residents and snowbirds (seasonal residents of 3-6 months)
regarding where they want to receive their information regarding Florida red tide and
ratings of trusted information sources regarding HAB information. This survey was a
subset of a larger survey investigating people decision making during a Florida red tide
(see Byrne and Studts abstracts). The survey was administered via SurveyMonkey in the
summer, 2012.
Results: The overall study population was predominantly older with a mean age (±SD) of
58.8 ± 13.6 years (range: 21-88 years), female (66.5%), and highly educated (60% had at
least a 4 year degree). When queried what 3 organizations people go to for information
regarding Florida red tide, participants selected private organizations (75%), media
(67%), and state agencies (65%). When queried about the type of information products
people prefer, they preferred websites (70%), television (56%), and newspapers (54%) as
the top three products. When queried about who they trust the most for Florida red tide
information (0 = no trust, 1 = A little, 2= Some, and 3 = A lot), private organizations were
trusted the most (2.84 + 0.45), with state and federal agencies the second and third
choices (2.53 + 0.69 and 2.46 + 0.75 respectively).
Discussion: Although websites were selected as the top mechanism people seek for
information regarding Florida, it should be noted that traditional sources such as media
and newspaper were also highly preferred. Private organizations are the most trusted
for information, with state and federal agencies also highly trusted. It should be noted
that the survey respondents were older and well educated. We suspect that the lack of
interest in social media such as Facebook and Twitter could be could be due to an age
cohort effect. The Mote Marine Laboratory sent the survey recruitment email, and so
could have caused positive bias toward private organizations such as Mote.
Conclusion: Our survey suggests that websites produced by private organizations are
the preferred information source for HAB information such as Florida red tide.
Page 162
STIMULATION OF DOMOIC ACID PRODUCTION FROM TRANSIENT
CHANGES IN NUTRIENTS
Raphael M. Kudela1, G. Jason Smith2, Kendra Hayashi1 and Clarissa Anderson1
Cruz, CA 95064, USA
2
Moss Landing Marine Laboratories, 8272 Moss Landing Rd, Moss Landing, CA 95039, USA
1
Page 163
S
October 2013
ORAL PRESENTATIONS
POSTER
There is a wealth of evidence from both laboratory and field settings that domoic
acid production in the genus Pseudo-nitzschia is linked to nutrient availability. Toxin
production is generally linked to physiological stress and/or growth rate, suggesting
that nutrient availability is simply modulating this response. Here we synthesize data
from a range of experiments including semi-continuous batch cultures, xenic and axenic
chemostats, and field studies (including both nutrient-amended grow-out experiments
and natural perturbations) to examine the transient changes in domoic acid production
resulting from changes in nutrients. While there is clear evidence for nutrient stress
(slowing or cessation of growth) as a cause for increased toxicity, we also see evidence
for rapid (days) changes in toxin cell quotas and particulate versus dissolved fractions,
triggered by changes in nutrient type and concentration. These rapid responses are
transient, indicative of physiological adjustments within the cellular biosynthetic
pathways. Domoic acid production also reaches a “steady state” in the chemostat
experiments after a greater delay than for other common metrics (cell density, pigment
per cell, C:N ratio). These shifts in toxicity in response to a variety of conditions, not just
nutrient stress leading to slower growth, perhaps explain the often contradictory reports
in the literature regarding the triggers for toxin production. We use these data to test
several of the recently proposed physiological models of domoic acid production that
have been developed. Based on concomitant measurements of variable fluorescence
(Fv/Fm), we suggest that Fv/Fm may be a universal indicator of cell toxicity, perhaps
providing an easy to use and cost effective method for tracking toxin potential that could
serve as a “trigger” for sampling using more conventional (but also more expensive and
time-consuming) methods or in conjunction with automated detection systems such as
the Environmental Sampling Processor (ESP).
SILICON CONTROL OF DIATOM COMPETITORS AGAINST THE RED TIDE
DINOFLAGELLATE, KAREnIA bREvIS, WITHIN THE EASTERN GULF OF
MExICO
Jason M. Lenes1, Brian P. Darrow1, John J. Walsh1, L. Kellie Dixon2, Gary J. Kirkpatrick2 and
Margie R. Mulholland3
College of Marine Science, University of South Florida, 140 7th Avenue South, St.
Petersburg, FL 33701, USA
2
3
Department of Ocean, Earth and Atmospheric Sciences, Old Dominion University, 5115
Hampton Boulevard, Norfolk, VA 23529, USA
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
The role of dissolved silicon was explored numerically in 2001 as a potential control
of annual phytoplankton succession, from diatoms to the toxic dinoflagellate, Karenia
brevis, observed in the eastern Gulf of Mexico each year during these and other field
samples of 1998-2001, 2003, and 2007-2011. Multivariate plots were constructed
during some of the latter years of the percent in situ biomass of the total phytoplankton
population attributed to each functional group of diatoms, cyanophytes, cholorophytes,
haptophytes, prasinophytes, K. brevis, other dinoflagellates, cryptophytes, and
prochlorophytes against each other and the dissolved inorganic and organic forms of
silica, nitrogen, and phosphorus macronutrients. They suggested that diatoms and K.
brevis were unlikely to co-exist. Diatom chlorophyll biomass off southwestern Florida,
from Apalachee Bay to the Ten Thousand Islands, did not exceed 1 μg chl l-1, when
silicate concentrations were <0.9 μmol SiO4 kg-1. Similarly, K. brevis abundances were
<40% of the total micro-algal population at concentrations of >3 μmol SiO4 kg-l when
K. brevis concentrations were <1 x 106 cells l-1. Within these niche constraints, the
percentage of K. brevis was highest at total dissolved nitrogen concentrations of 6-14
μmol N kg-1 and at total dissolved phosphorus concentrations of 0.2-0.8 μmol P kg-1.
The impact of Si-limitation was quantitatively studied with a one dimensional causal
simulation model of the 2001 K. brevis harmful algal bloom (HAB) on the West Florida
Shelf (WFS). These model results indicated that just alleviation of Si-limitation among
diatoms reduced the depth-integrated biomass of competing HABs of K. brevis by 36%
over the 20-m water column.
Page 164
GAmbIERdISCuS NUTRIENT UPTAKE KINETICS
Alexander K. Leynse and Michael L. Parsons
Florida Gulf Coast University, 10501 FGCU Boulevard South, Fort Myers, FL 33965, USA
Ciguatera fish poisoning (CFP) is the most wide-spread phycotoxin-borne illness in
the world, responsible for tens of thousands of poisonings per year world-wide. The
causative toxins are produced by epibenthic dinoflagellates of the genus Gambierdiscus.
These toxins bio-accumulate and biomagnify in tropical fish causing these fishes to
become toxic for human consumption.
CFP outbreaks have been difficult to predict, due in part to taxonomic ambiguities
and an incomplete understanding of ecological aspects of Gambierdiscus. With recent
revisions in Gambierdiscus taxonomy, it is important that future research focuses on
ecophysiological, toxicological, and host preference variation among Gambierdiscus
species and strains.
Many observed increases in harmful algal blooms appear to be related to anthropogenic
nutrient loading. However, the influence of nutrient concentrations on Gambierdiscus
blooms and toxin production are unclear. Several field studies have been conducted,
but oftentimes Gambierdiscus cells densities did not correlate with inorganic nutrient
concentrations. It is likely that species variation plays some role in this disconnection.
Page 165
S
October 2013
ORAL PRESENTATIONS
POSTER
In this study, we measured the nitrate and ammonium uptake rates of multiple
Gambierdiscus species in different concentrations of each nutrient. These data were
then used to construct Michaelis-Menten kinetic equations. Species comparisons will
be discussed as well as the future application of these data to ecological models for
predicting ciguatera outbreaks.
CHANS: FRAMING THE RED TIDE STORY: DAILY NEWSPAPER COVERAGE
IN SOUTHWEST FLORIDA
Z. Li1, B. Garrison1, and S. Ullmann2
School of Communication, University of Miami, Coral Gables, FL 33146
1
Coral Gables, FL 33146
2
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
One of the central themes in media effects studies is the agenda-setting role of mass
media, which argues that the mass media impact heavily on the dominant issues being
discussed in the public agenda. Furthermore, as part of the efforts of agenda-setting
research, scholars have argued over the “framing” effects of mass media, which draws
attention to how particular news coverage attributes dominate – or frame – the way
certain topics are addressed in storytelling. Based on framing theory, this study focuses
on news media reporting about HABs (specifically Florida red tide) along the Southwest
Coast of Florida during the past two decades.
The study examines how the red tide story has been framed by the media (e.g., as a
human health risk issue, an environmental story focused on marine animal threats, a
business story emphasizing tourism and recreation, and so forth). Approximately 20
years of data are content analyzed using the NewsBank and Lexis-Nexis databases from
four local daily newspapers in three standard metropolitan statistical areas defined
by the U.S. Census Bureau along the Florida Gulf Coast: Sarasota-Bradenton, Tampa
Bay-St. Petersburg, and Bonita Springs-Naples. A coding scheme is designed based on
previous literature and pretested for reliability. Two coders are used to collect data. The
variables examined include the theme or frame of news storytelling, story type, the
sources journalists use to report the story, and images used to supplement storytelling.
Other newspaper story characteristics are also measured. We are also interested in how
the health-related information has been communicated by the news coverage. Finally,
the study looks at the coverage findings in the context of hospital occupancy, regional
tourism, and timing of active red tide blooms. The study findings offer insights as to the
long-term outcome of the public awareness and perceptions of Florida red tide.
Page 166
HABITAT MODELING OF PSEudO-nITZSCHIA DISTRIBUTION AND
TOxICITY IN THE COASTAL WATERS OF THE NORTHWEST PACIFIC USING
NON-PARAMETRIC MULTIPLICATIVE REGRESSION
Susan C. Lubetkin1 and Evelyn J. Lessard1
University of Washington, School of Oceanography, Box 357940, Seattle, WA 98195,
USA
1
It has been difficult to assess the environmental regulators of toxigenic Pseudo-nitzschia
spp. blooms within the Juan de Fuca eddy and along the Pacific Northwest coast using
traditional statistical methods, perhaps in part because of non-linear interactions
between important variables. We are applying a new statistical habitat modeling
method, non-parametric multiplicative regression (NPMR), to the rich dataset collected
over ten ECOHAB and RISE cruises from 2003 to 2006 to isolate factors which best
describe the abundance of Pseudo-nitzschia spp. and domoic acid (DA) production.
NPMR combines predictor variables multiplicatively, such that the effect of one predictor
variable can covary in a complex way with others, and requires no assumptions about
the overall shape of the response surface. Using NPMR, we isolated several variables
from a list of over 40 physical, chemical, and biological variables, which in combination
are good predictors of Pseudo-nitzschia spp. and DA concentrations. The list of
predictors varied across sampling subsets (i.e, season, cruises with extremely high
Pseudo-nitzschia spp. concentrations), giving us new insights into the complex factors
leading to toxigenic Pseudo-nitzschia spp. blooms in this region.
ORAL PRESENTATIONS
POSTER
Page 167
S
October 2013
IDENTIFICATION OF CYANOBACTERIAL TOxINS INVOLVED IN THE
APPARENT INTOxICATION OF DOLPHINS IN THE FLORIDA KEYS
Christina Lydon1, Shunmei Liu2, Larry Brand3 and John P. Berry1
International University, 3000 NE 151st Street, North Miami, FL 33181
2
Department of Pharmaceutical and Biological Science, Weifang Medical University, 7166
West Baoton Street, Weifang, Shandong Province, 261053
3
Sciences, University of Miami, 4600 Rickenbacker Causeway, Miami, FL 33146
1
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Cyanobacteria produce a diverse repertoire of toxic or otherwise bioactive metabolites
that have been linked to intoxication of humans and wildlife. Captive dolphins in
the Florida Keys were recently observed to graze cyanobacteria, and perhaps other
associated algae, occurring within enclosures followed by apparent signs of intoxication,
and specifically seeming neurotoxicity. To identify possible toxic metabolites involved
in these intoxication events, we have employed the zebrafish (Danio rerio) embryo, as
a model of vertebrate toxicity, toward bioassay-guided fractionation/purification, and
subsequent chemical and toxicological characterization, of relevant biologically active
metabolites. Initial studies indicate that extracts, and several subsequent chemical
fractions, derived from cyanobacteria are, indeed, toxic in the zebrafish model, and
generally support these as potentially toxic algae. Current progress on the purification
and characterization of these compounds will be presented.
Page 168
POTENTIAL HAB SPECIES FOUND IN THE CALOOSAHATCHEE ESTUARY,
FLORIDA
Laura R. Markley and Michael L. Parsons
Coastal Watershed Institute, Florida Gulf Coast University, 10501 FGCU Blvd South, Fort
Myers, FL, United States 33965
Page 169
S
October 2013
ORAL PRESENTATIONS
POSTER
Anthropogenic activities over the last several decades are thought to play a role in the
increased frequency, duration and intensity of harmful algal blooms (HABs) worldwide.
While the dominant HAB species in the Gulf of Mexico, Karenia brevis, continues to be
a focus of scientific study, research has been sparse on estuarine phytoplankton and
other potential HAB species that may be present in inshore environments. In southwest
Florida, the Caloosahatchee is a highly managed riverine system where freshwater
inflow can be negligible in the dry season (winter-spring) and extremely high (>5000
cfs) in the wet season (summer-fall). As freshwater inflow is a primary controlling factor
of both nutrient and light availability for phytoplankton in the Caloosahatchee estuary,
phytoplankton growth can vary greatly throughout the year. We collected monthly
plankton samples from 2008 to 2010 at 14 locations along the Caloosahatchee estuary
from the Franklin lock and dam (S-79) to the river mouth. Phytoplankton were identified
to the lowest taxonomic level possible and quantified as cells/L and cell volume/L.
Several previously undocumented HAB species were found in the estuary (e.g., Akashiwo
sanguinea and Pseudo-nitzschia pungens). Blooms were evident year-round, occurring
upstream when freshwater inflows were low and further downstream as rates of
discharge increased. As several species documented by this study have caused HABs
in other areas, further studies should be conducted to determine if the strains of the
species found in the estuary are toxic and what environmental conditions promote cell
growth. These species also need to be considered in the management of freshwater
inflow from S-79 in order to minimize the threat of HABs in the Caloosahatchee estuary.
TIME SERIES ANALYSIS FOR PERIODS WITH HIGH CONCENTRATION
VERSUS PERIODS WITH LOW CONCENTRATION OF KAREnIA bREvIS
Grace Maze1, Josefina Olascoaga1, Larry Brand1
1
University of Miami, RSMAS, 4600 Rickenbacker Causeway, Miami, FL, 33146
The West Florida Shelf is home to a toxic dinoflagellate, Karenia brevis. This
dinoflagellate produces brevetoxins, a suite of polyether neurotoxins, which are
hazardous to mammals and marine life. Since 1954, 28 K. brevis blooms with a
concentration >104 cells/L have been observed off the coast of west Florida between
Tampa Bay and Charlotte Harbor. There are many theories on what causes the onset of a
bloom, however no theory currently accounts for all blooms.
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Using Karenia brevis concentration between 26.5oN and 28oN and shoreward of 85oW
periods of no bloom (<100 cells/L) and periods of large bloom (>105 cells/L) were
identified. For each case the concentration levels were required to continue for at least
10 days with no deviations lasting more than 5 days. From 1992 to 2006, 15 instances
were found with no bloom detected and 8 instances were detected with a large bloom.
Values for alongshore wind, cross shore wind, Peace River discharge, Caloosahatchee
River discharge, and the northern most position of the Loop Current were analyzed
to compare the periods of no bloom versus the periods of large blooms. Using a twosided student’s t-test the average values were compared and it was found that the only
statistically significant difference between periods of large bloom and periods of no
bloom was the freshwater discharge of the Peace River.
Page 170
DEVELOPMENT OF COMPETITIVE FLUORESCENCE-BASED BINDING
ASSAYS FOR BREVETOxINS AND BREVENAL
Jennifer McCall, Karl Jacocks, Allan Goodman, Susan Niven, Alysha Thompson, Daniel
Baden, and Andrea Bourdelais
Center for Marine Science, UNCW, 5600 Marvin K. Moss Lane, Wilmington, NC 28409
Page 171
S
October 2013
ORAL PRESENTATIONS
POSTER
Brevetoxins are a family of ladder-frame polyether toxins produced during blooms of
the marine dinoflagellate Karenia brevis. Consumption of fish exposed to K. brevis
blooms can lead to the development of neurotoxic shellfish poisoning. The toxic effects
of brevetoxins are due to activation of voltage-sensitive sodium channels (VSSCs) in
cell membranes. K. brevis also produces a non-toxic antagonist to brevetoxin known
as brevenal, which is able to inhibit and/or negate many of the detrimental effects of
brevetoxins. Brevenal binding to VSSCs has yet to be fully characterized, in part due
to the difficulty and expense of current techniques. Binding of toxins has historically
been measured using a radioligand competition assay that is fraught with difficulty. In
this study, we developed two novel fluorescence-based binding assays (FBAs) for the
brevetoxin receptor and the brevenal receptor. Several fluorophores were conjugated
to PbTx-2 or brevenal and used as the labeled ligands. Both assays were qualified
against the standard radioligand receptor assay for the respective receptors and
yielded comparable equilibrium constants. On the brevetoxin FBA, brevetoxin analogs
were able to compete for binding with the fluorescent ligands. The FBA was used to
determine relative concentrations of toxins in raw extracts of K. brevis culture, and to
determine ciguatoxin affinity to site 5 of VSSCs. Using the brevenal FBA, in depth studies
to characterize brevenal’s binding site on rat brain synaptosomes have shown that the
fluorescent brevenal conjugate was not displaced by VSSC ligands for sites 1-5 or 7,
or the sodium channel blocker amilioride, suggesting that brevenal does not bind to
VSSCs at any of these locations. As such, brevenal may elicit its action through a novel
mechanism and/or currently unknown receptor site on VSSCs. The FBAs are quicker,
safer, far less expensive, and do not generate radioactive waste or need radioactive
facilities. As such, these assays can be used to replace the current radioligand assays
and will be a vital tool for future experiments examining the binding affinity of various
ligands on sodium channels.
DEVELOPMENT OF PCNA AS A MOLECULAR BIOMARKER FOR GROWTH
STATUS OF KAREnIA SPECIES IN GULF OF MExICO RED TIDES
Megan E. Meek1,2, Alina Corcoran3, and Frances M. Van Dolah2,1
Graduate Program in Marine Biology, College of Charleston, Charleston, SC 29412
3
St. Petersburg, FL 33701
1
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
2
Blooms of the toxic dinoflagellate Karenia brevis have been documented in the Gulf
of Mexico for more than 125 years. There have been many efforts to model K. brevis
blooms to forecast impacts and improve management practices. Current models
focus on physical drivers of large scale bloom dynamics, but do not provide insight
into biological processes occurring within a given bloom patch that may be predictive
of its fate. Moreover, models focus solely on the single species K. brevis, ignoring cooccurring Karenia mikimotoi, Karenia selliformis, and Karenia papilionacea, each of
which has unique temporal and spatial distributions relative to K. brevis blooms. The
purpose of this study is to develop a molecular biomarker for K. brevis growth rate and
adapt it for use with existing Karenia probes to enhance monitoring and inform model
development. The proliferating cell nuclear antigen (PCNA) protein is part of the DNA
replication fork complex that occurs during the DNA synthesis (S) phase of the cell cycle.
In K. brevis, the cell cycle is under circadian control. PCNA is present throughout the cell
cycle in K. brevis, but its expression increases in actively growing cells, with the highest
concentration occurring during S-phase. In laboratory cultures, a linear correlation is
observed between growth rate and expression of PCNA (assessed by western blotting),
suggesting a potential for PNCA as a biomarker for growth rate. Here we present
preliminary data on the comparative circadian cell cycle behavior and PCNA expression
in Karenia species from the Gulf of Mexico. We demonstrate that the anti-K. brevis PCNA
antibody recognizes PCNA not only in K. brevis, but also in other Karenia species. We are
currently adapting PCNA detection to a flow cytometric assay suitable for monitoring
applications, with the application of species-specific rRNA probes in conjunction with
anti-PCNA detection to enable the simultaneous determination of Karenia species
composition and relative growth in mixed bloom populations.
Page 172
LOCALIZATION OF BREVISIN-DAUNORUBICIN CONJUGATE IN SJCRH30
CELLS
Nina E. Neill, Elizabeth A. Elliott, Allan J. Goodman, Daniel G. Baden and Andrea J.
Bourdelais
Center for Marine Science University of North Carolina at Wilmington, Wilmington NC
28409
Page 173
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ORAL PRESENTATIONS
POSTER
One of the most difficult challenges facing drug discovery is the issue of maximizing
the bioavailability of active pharmaceutical agents, which is in part regulated by a
compound’s ability to cross cellular membranes and overcome efflux mechanisms within
the cell. In an effort to overcome these barriers, increasingly sophisticated methods to
improve a drug’s effectiveness are being pursued, such as stimuli specific drug delivery
systems and bio-engineered nanomolecular carriers. Research at UNCW’s Center for
Marine Science on ladder frame polyethers (LFPs), including brevenal, brevisin and
brevetoxins, produced by the dinoflagellate Karenia brevis, has led to another possible
drug carrier with the capability of increasing the bioavailability of compounds that
normally exhibit poor cellular uptake. LFPs demonstrate an ability to rapidly cross cell
membranes. When conjugated to large compounds with low membrane permeability,
such as the chemotherapeutic agent daunorubicin, LFPs retain the ability to quickly cross
the cell membrane while concurrently escorting the drug into the cell. Once inside the
cell, the LFP-drug conjugates localize in discrete subcellular compartments, the identity
of which is critical to the complex’s function. By staining several cellular organelles and
visualizing the uptake and localization of the naturally fluorescent brevisin-daunorubicin
compounds, we were able to determine that the LFP conjugates primarily localize in the
endoplasmic reticulum as well as the late endosomes. In contrast, daunorubicin alone
has previously been shown to sequester primarily in lysosomes as well as the nucleus, to
a lesser extent (Schindler 1996). Lysosomes are the enzyme containing organelles that
are responsible for cellular waste degradation, leading to subsequent expulsion from
the cell. Many basic drugs have been shown to accumulate within these organelles,
which inhibits their efficacy. Our results have implications for future drug delivery
developments as the property of LFP-conjugates to quickly and efficiently traverse
cellular membranes and localize in new organelles could be capitalized upon to increase
pharmaceutical bioavailability.
RESPONSES OF A NATURAL KAREnIA BLOOM AND OTHER TAxA OF THE
WEST FLORIDA SHELF TO SINGLE NUTRIENT, MULTIPLE NUTRIENT, OR
ESTUARINE WATER ADDITIONS
Ari Nissanka1, L. Kellie Dixon1, Susan Launay1, Lori J. Zaworski1, Patricia J. Minotti1, Camia
M. Charniga1, Alexandria G. Hounshell1, Jennifer M. Vreeland1
POSTER
PRESENTATIONS
TIONS ORAL
PRESENTA
1
While blooms of the toxic dinoflagellate, Karenia brevis, predominantly originate in
oligotrophic waters offshore and while K. brevis is slow-growing relative to the diatoms
that form a dominant community of the coastal phytoplankton on the West Florida Shelf,
K. brevis blooms are carried inshore where they can outcompete other species and form
nearly monospecific concentrations of many millions of cells per liter. The potential
sources of nutrients to maintain such blooms are varied, but include estuarine outflow
from the three major estuaries of SW Florida. A program was initiated to challenge and
quantify the response of a naturally occurring K. brevis bloom with a variety of nutrients,
including water from the three estuaries. A larger scale (~20 L per sample) bioassay
was conducted with a natural bloom of 1.0-2.6 X 105 cell L-1. Treatments included three
amino acids, inorganic nitrogen species (ammonia, nitrate), urea, inorganic phosphorus,
silicate, urea plus inorganic phosphorus, and individually, the reduced salinity waters
of the three major local estuaries. Reductions in salinity as a result of either reagent
or estuarine water additions were carefully matched. Treatments were randomly
prepared in triplicate, and samples incubated in ambient water at the surface, but
under reduced light intensity (60%) for approximately 48 hours. Similar to prior work
on coastal phytoplankton communities in the absence of Karenia, nitrogenous additions
displayed responses in K. brevis growth and total biomass, elemental ratios, and other
phytoplankton community components that differed in trajectory and endpoints from
those of the other treatments. There was little response to the estuarine additions.
The urea plus inorganic phosphorus resulted in the highest growth rates of Karenia,
above that of either urea or phosphorus alone and with the lowest consumption of
nitrogen. Among treatments, similar amounts of total fixed nitrogen were achieved
with very different nitrogen uptakes due to the success of other taxonomic groups. In
an interesting interaction between K. brevis and diatoms, the highest silica depletion
was observed in the urea plus phosphorus treatment that also experienced the highest
K. brevis growth rates. Additional combined treatments (N+Si, Urea+Si+P) should be
investigated.
Page 174
ISOLATION AND CHARACTERIZATION OF A BREVENAL EPIMER FROM
CULTURED KAREnIA bREvIS
Susan Niven1, Andrea Bourdelais1, Jen McCall1, Alysha Thompson1, Karl Jacocks1, Ned
Martin2, Bill Abraham3, and Daniel Baden1
Center for Marine Science, University of North Carolina at Wilmington, Wilmington NC
28409
2
Department of Chemistry and Biochemistry, University of North Carolina at Wilmington,
Wilmington NC 28403
3
Division of Pulmonary and Critical Care Medicine, University of Miami at Mount Sinai
Medical Center, Miami Beach, FL
1
Cell based, receptor binding and in vivo experiments have shown that the small
difference in structure has a dramatic effect on the receptor binding to the brevenal and
brevetoxin receptors. Epi-brevenal showed lower affinity for the brevenal receptor (Ki
epi-brevenal = 2025 nM vs Ki brevenal = 146 nM ), however epi-brevenal had increased
affinity for the brevetoxin receptor (Ki epi-brevenal = 3.1 nM vs Ki brevenal = 1133 nM).
Both brevenal and epi-brevenal did not increase bronchoconstriction in sheep on their
own and blocked brevetoxin induced bronchoconstriction significantly.
These results indicate that the stereochemistry at C27 is crucial for optimal affinity of
brevenal to its receptor and suggest that the final ring closure is not enzymatic but
chemical in nature, with the lowest energy conformer as the major product.
Page 175
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October 2013
ORAL PRESENTATIONS
POSTER
Brevenal is a ladder frame polycyclic ether compound produced by Karenia brevis that
was first described in 2004. Research has identified brevenal as a natural antagonist
to the brevetoxins which can facilitate mucus clearance in mammals and block
brevetoxin activity. Since its discovery, isolation methods for brevenal from K. brevis
have been optimized to improve yield and increase purity. During the development of
these methods, a second compound with a similar retention time and identical mass
(MW=656.4) as brevenal was isolated. Comparison of 1H NMR spectra with brevenal
showed large areas of similarity, but did not overlay exactly. A full suite of 1D and
2D NMR experiments were performed and the structure was elucidated. The new
compounds was structurally similar to brevenal, but the chemical shifts of protons and
carbons on the ladder frame were different starting at C18, and showed maximum
differences in chemical shifts occurring at C27. Complete structure determination
identified the new compound as an epimer of brevenal with different stereochemistry
at C27. The other chemical shifts observed in the 1H NMR spectrum (C18-C26) are likely
due to conformational changes induced by the different connectivity at C27. Molecular
modeling comparisons of brevenal to epi-brevenal suggest that brevenal is in the lowest
energy conformation with epi-brevenal at a slightly higher energy level.
ARE FATTY ACID AMIDES RESPONSIBLE FOR FISH MORTALITY DURING
PRymnESIum PARvum BLOOMS?
Sean P. O’Mara1, Greg Southard2, Danielle Gutierrez1, and Paul V. Zimba1
Texas A & M University – Corpus Christi, 6300 Ocean Drive, Corpus Christi, TX 78412,
USA
2
Texas Parks and Wildlife Department, Inland Fisheries-Analytical Services, 507 Staples
Road, San Marcos, TX 78666, USA
ORAL
POSTER
PRESENTATIONS
PRESENTATIONS
1
The global frequency and distribution of harmful algal blooms has increased in recent
decades. One species of concern is Prymnesium parvum - a microscopic, unicellular,
flagellate from the phylum Haptophyceae, which has been associated with harmful
algal blooms for nearly a century. The primary toxins produced by P. parvum have long
been thought to be two metabolites, prymnesin 1 and 2. However, it was recently
shown that toxic fatty acid amides (FAA) were undescribed toxic compounds produced
by P. parvum. The purpose of this study will be to (1) determine a correlation of FAA
concentration to P. parvum cell number using at least 5 strains isolated from sites
distributed worldwide, and (2) perform fish dosing studies to determine the effect of pH
on FAA toxicity. Fish dosing studies will be conducted in a full factorial design utilizing
2 FAA concentrations and 3 pH values. Unlike previous studies, preliminary findings
suggest a strong correlation (r2=.87) between FAA concentration and cell number of field
samples collected in Texas river systems, and a moderate correlation (r2=.33) between
FAA concentration and ichthyotoxicity. A multiple regression analysis will be conducted
to determine the environmental conditions (e.g. pH, salinity, temperature, etc.) that
influence FAA concentration and ichthyotoxicity in Texas field samples. In addition,
preliminary analysis of P. parvum strains supports previous findings that toxin production
is reduced or halted during times of excess nutrients.
Page 176
PHOTOOxIDATION OF DOMOIC ACID
Sk Md Sazzad Hossain1, Jenna L. Logsdon1 Dionysios D. Dionysiou2, Kevin E. O’Shea1
Department of Chemistry and Biochemistry, Florida International University, Miami, FL,
33199, USA
2
Environmental Engineering and Science Program, University of Cincinnati, Cincinnati, OH
45221-0012, USA
1
Domoic acid, a structural analog of kainic acid, is a potent neurotoxin produced by the
diatom species Pseudo-nitzschia during harmful algal blooms (HABs). Consumption of
domoic acid tainted seafood leads to amnesic shellfish poisoning which causes shortterm memory loss, brain damage and in severe cases, death. While photooxidation may
play an important role in the transformation of domoic acid in the environment, the
photooxidation of domoic acid has received little attention. In the presence of sunlight,
oxygen and the colored pigments associated with HABs photodynamic action can lead
to the formation of singlet oxygen and transformation of a variety of naturally occurring
toxins. Domoic acid contains a number of functional groups, including a conjugated
diene and carboxylic acids. Because of the high cost of domoic acid we have chosen to
study the singlet oxygenation of sorbic acid, which also contains a conjugated diene and
carboxylic acid functional groups as a model compound for domoic acid.
Our results demonstrate sorbic acid readily undergoes singlet oxygenation yielding a
complex mixture of peroxide type products. We will report detailed product studies for
the photooxidation of sorbic acid based on one and two-dimensional NMR techniques.
The reaction kinetics and bimolecular rate constant for sorbic and domoic acids in
aqueous media were evaluated using competition kinetics with furfuryl alcohol. The
results of ongoing product studies and toxicity tests as a function of photooxidation of
domoic acid will be reported.
POSTER
ORAL PRESENTATIONS
October 2013
S
Domoic Acid
Sorbic Acid
Page 177
BIOACCUMULATION OF BREVETOxINS AND MAJOR METABOLITES
IN FILTER-FEEDING AND CARNIVOROUS MOLLUSKS ExPOSED TO
NATURAL KAREnIA bREvIS HARMFUL ALGAL BLOOMS
R. Pierce1, M. Henry1, P. Blum1, R. Medvecky1, L. Flewelling2
1
ORAL
POSTER
PRESENTATIONS
PRESENTATIONS
2
Harmful algal blooms (HABs) of the dinoflagellate Karenia brevis (Florida red tide) occur
annually along the Florida Gulf coast. Neurotoxic compounds (brevetoxins, PbTxs) produced
by K. brevis are ingested by filter-feeding mollusks as they filter K. brevis cells and toxins
from the water. Human consumption of contaminated shellfish results in neurotoxic
shellfish poisoning (NSP) that can cause serious neurological and gastrointestinal problems.
Multiple cases of NSP have resulted from human consumption of various filter-feeding and
well as carnivorous mollusks harvested non-commercially from unapproved areas. This
study was undertaken to investigate the accumulation, transformation and persistence
of brevetoxins and selected metabolites within various species of estuarine mollusks
that are likely vectors of NSP. The most abundant metabolites observed to accumulate
in filter-feeding mollusks (clams and oysters) were cysteine conjugates of brevetoxins.
Parent brevetoxins produced by K. brevis (PbTx-1,-2,-3, etc.) were not detected in filterfeeding mollusks, except for low concentrations of PbTx-3. Filter-feeding mollusks retained
the PbTx metabolites for several months after HAB termination, but did not retain NSP
toxicity. The carnivorous mollusk (lightning whelk) exhibited NSP toxicity 6 months after
HAB termination. Elevated concentrations of PbTx-3 and metabolites were still present
when the study ended 7 months after HAB termination.
Page 178
GROWTH AND TOxIN PRODUCTION OF THE MIxOTROPHIC
DINOFLAGELLATE ALExAndRIum CATEnELLA
Regina Radan1 and Raphael M. Kudela1
1
Page 179
S
October 2013
POSTER
ORAL PRESENTATIONS
Despite the role of inorganic nutrients in maintaining toxic blooms of the dinoflagellate
Alexandrium catenella, there is strong evidence for use of both particulate ingestion
(mixotrophy) and direct utilization of organic compounds. A. catenella causes
detrimental effects to humans, marine mammals, fish and seabirds from ingestion
of contaminated fish or shellfish, and is routinely monitored in California because of
the potential for negative impacts at low cell densities. Mixotrophy has the potential
to provide A. catenella a competitive advantage over other phytoplankton when in
nutrient-limited conditions. Another common strategy employed by dinoflagellates is
to vertically migrate to acquire inorganic nutrients at depth. It is unclear whether A.
catenella utilizes both strategies, and if so, whether these strategies are complementary,
and how these nutritional modes might change growth and toxicity in California strains
of A. catenella. Preliminary data has shown that cultures of A. catenella grown on a
base medium of f/2 and then starved will ingest the haptophyte Isochrysis galbana.
We will present results for growth, toxicity, and nutrient utilization using a laboratorybased vertical migration chamber to assess whether A. catenella exhibits a preference
for vertical migration or mixotrophy when presented with both potential modes of
nutrition. Cells in nutrient replete conditions should maintain faster growth rates then
cells in nutrient limited conditions. If prey is present in nutrient-replete conditions A.
catenella may ingest I. galbana as an additional nutrient source. However, in nutrientlimited conditions, I. galbana should allow A. catenella to maintain growth but won’t
necessarily increase growth rate. Given the tradeoff between energy expenditure on
vertical migration and mixotrophy, we expect there to be significant difference in cellular
toxin quota due to changes in cellular stress and metabolic balance. These results will
ultimately lead to a better understanding A. catenella growth dynamics and could be
used to improve mechanistic models of growth and toxin production.
QUANTIFICATION OF DOMOIC AND OKADAIC ACID IN SHELLFISH AND
ALGAE USING A LC/MS/MS (TRIPLE QUADRUPOLE) APPROACH
Carlton Rauschenberg1, Craig Burnell1 and Stephen Archer1
1
Bigelow Laboratory for Ocean Sciences
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
A series of test results are presented for the quantification of amnesic shellfish poisons
(ASP) and diarrhetic shellfish poisons (DSP) in shellfish. The previously described LC/
MS/MS approach (McNabb et al 2005 [M2005]) is practical for use in routine monitoring
programs and the test results shown represent the initial steps towards single laboratory
validation to quantify ASPs, DSPs and other lipophilic algal toxins in shellfish at Bigelow
Laboratory. A comparison between the M2005 approach and traditional LC-UV approach
is presented for domoic acid, addressing sensitivity and specificity of detection in
shellfish matrix-fortified standards. The M2005 approach is applied to explore seasonal
variations in both ASP and DSP content in natural shellfish populations in the Gulf of
Maine.
Page 180
USE OF DISSOLVED INORGANIC AND ORGANIC NITROGEN BY THE
TOxIC DINOFLAGELLATES, KAREnIA bREvIS AND KAREnIA mIKImOTOI
(DINOPHYCEAE)
Bill Richardson
100 8th Avenue SE, Saint Petersburg, FL 33701, USA
Page 181
S
October 2013
ORAL PRESENTATIONS
POSTER
Dissolved inorganic nitrogen (DIN) and dissolved organic nitrogen (DON) compounds
are important sources of nitrogen (N) for large harmful blooms of Karenia brevis and
Karenia mikimotoi. The ability to use different DIN and DON compounds as sole sources
of N for growth was evaluated in the laboratory using two Karenia brevis clones (CCFWC
257, CCMP 2281) and a single Karenia mikimotoi clone (CCFWC 67) grown axenically
and non-axenically on forty-nine different N-containing compounds. DIN compounds
included NH4+, NO3-, and NO2- and DON compounds included amino acids, peptides,
a protein, nucleic acids, nucleotides, purines, a polyamine, and an amino sugar. Both
axenic and non-axenic cultures grew well on all DIN compounds, but differed in their
ability to use DON. Axenic cultures of K. brevis and K. mikimotoi were unable to grow on
any DON compound other than urea. In contrast, non-axenic cultures of K. brevis and
K. mikimotoi grew well on urea and most of the other DON compounds. Among the N
compounds that supported growth, no single compound provided exceptionally superior
growth for either axenic or non-axenic Karenia cultures. Overall, growth was better on
DIN compounds than on DON compounds. Differences in growth between the axenic
and non-axenic cultures indicated that K. brevis and K. mikimotoi were able to directly
use all DIN forms and urea for growth, while the use of other forms of DON for growth
was conditional upon prior bacterial mineralization. The results suggest a potentially
beneficial relationship in which N-rich components of the DON pool not directly
accessible to either K. brevis or K. mikimotoi are mineralized by co-occurring bacterial
communities into N forms that can then be used by these Karenia species to meet their
cellular N growth demands. This relationship may become increasingly important for
both Karenia species when DIN and urea are in short supply.
THE EFFECTS OF FIELD ExPOSURE OF CRASSOSTREA vIRGInICA TO
KAREnIA bREvIS: HISTOPATHOLOGY AND BREVETOxIN ACCUMULATION
IN GAMETES
Anne Rolton1,2, Julien Vignier1,2, Audrey Barbe1, Philippe Soudant2, Richard Pierce3,
Sandra Shumway4, Monica Bricelj5 and Aswani Volety1.
Florida Gulf Coast University, College of Arts and Sciences, 10501 FGCU Blvd South, Fort
Myers, Florida 33965; 2Universitéde Bretagne Occidentale—IUEM, LEMAR CNRS UMR
6539, Place Nicolas Copernic, Technopôle Brest Iroise, 29280 Plouzané, France; 3Mote
Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236; 4Department
of Marine Sciences, University of Connecticut, 1080 Shennecossett Road, Groton, CT
06340; 5Institute of Marine and Coastal Sciences, Rutgers University, 71 Dudley Road,
New Brunswick, NJ 0890.
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
The most prevalent HAB species in the Gulf of Mexico is the red-tide dinoflagellate,
Karenia brevis, a producer of lipophilic brevetoxins (PbTxs). As well as impacting
commercially and ecologically important shellfish species, K. brevis is also a risk to
human health, with consumption of contaminated shellfish leading to neurotoxic
shellfish poisoning (NSP). The specific effects of K. brevis on shellfish reproduction
remain poorly understood; however, it is known to cause recruitment failure in some
shellfish species. The frequency of Karenia blooms is increasing and for certain shellfish
species, periods of ripening/spawning overlap with red-tide blooms. It is therefore
important to determine effects on reproduction and recruitment.
In Lee county, FL, the 2012-2013 red-tide began in September/October 2012 and has
continued through April 2013. The linear distance of the bloom has extended beyond
Lee county borders, about 100 miles from Lee county to Pinellas, FL. Throughout this
time, the eastern oyster, Crassostrea virginica, which contributes markedly to the
ecology of the region, has been exposed to the bloom during a time of gamete ripening,
spawning and larval recruitment. Although C. virginica can accumulate brevetoxins, it is
not known if there is maternal transfer of brevetoxin to the gametes. Previous laboratory
exposures of C. virginica to cultured K. brevis have shown increased histopathological
lesions in exposed oysters.
In this study, ripe adult eastern oysters were collected from a site exposed to K. brevis
(Iona cove, Caloosahatchee river, FL) and a site 200 miles north with no previous
exposure to K. brevis (Shellfish Harvesting Area 3402, Cedar key, FL). Oysters from
exposed and non-exposed sites were examined for histopathology and brevetoxin
content was assessed on stripped, filtered and concentrated gametes using LC-MS/MS
analysis.
Page 182
KAREnIA bREvIS REFERENCE TRANSCRIPOME ASSEMBLY AND
TRANSMEMBRANE PROTEIN SEQUENCE DISCOVERY
Darcie Ryan1 and Lisa Campbell1
Texas A&M University, 3146 TAMU Dept. Oceanography, College Station, TX 77843
1
Karenia brevis, a brevetoxin-producing dinoflagellate, blooms nearly annually in the
Gulf of Mexico and is linked to marine animal mortalities and human health threats.
Though the biological function of brevetoxins is unknown, elevated brevetoxin levels
in K. brevis cells have been measured during laboratory osmotic stress treatments.
To investigate the mechanisms underlying K. brevis toxin production, we assembled
reference transcriptomes for three clones: Wilson CCFWC268, SP3, and SP1, a low toxin
producing variant. These are among the first dinoflagellate transcriptomes assembled.
Each contained ≈90,000 contigs.
Complete core eukaryotic genes were identified in the transcriptomes to assess
completeness of assembly without a reference genome, and approximately 40% of the
contigs produced a significant hit (E value < 1.00E-6) with a sequence in the NCBI nonredundant (nr) protein database with a BLASTX search. Of the non-hits, more than 85%
contained a predicted complete open reading frame (ORF) that was ≥ 100 amino acids
long. These contigs represent potential unique K. brevis proteins.
Of the 32 putative ion channels, 16 were Na+ channels, 4 were Ca2+ channels, and 12
were K+ channels, based on BLASTx top hit results and conserved amino acid motifs.
They were identified with strict search criteria: each first hit one or more proteins in
a custom multi-species voltage-gated Na+ channel α subunit database, contained a
complete ORF and six predicted transmembrane regions, and finally significantly (E value
< 1.00E-6) contained one of the Pfam Ion_trans families during a conserved domain
search. The 32 putative ion channel sequences therefore represent a conservative set
that was chosen based on similarity to annotated voltage-gated Na+ channel units.
The identification of ion channels and aquaporins suggests that the concentrations of
ions and water within K. brevis cells are regulated by transmembrane proteins. Further,
K. brevis osmoacclimatory and osmoregulatory processes may be sensitive to voltage
gradients.
Page 183
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October 2013
ORAL PRESENTATIONS
POSTER
Putative aquaporin and voltage-gated Na+, Ca2+, or K+ channel sequences were
discovered in the transcriptomes using a variety of techniques, including conserved
domain searches and transmembrane region prediction. The putative aquaporin
sequence was 405 aa long and highly conserved among clones. The sequence very
significantly (E value < 1.00E-20) matched over 70 aquaporin or potential aquaporin
entries in the nr database. However, the top three hits were hypothetical proteins from
Aureococcus anophagefferen, Emiliania huxleyi, and Guillardia theta. This indicates that
similar major intrinsic proteins are present in algae other than K. brevis, but have not yet
been annotated.
BIOFOULING TUNICATES ON AQUACULTURE GEAR AS POTENTIAL
VECTORS OF HARMFUL ALGAL INTRODUCTIONS
M. Rosa1, S.E. Shumway1, B.A.Holohan1, S.G. Bullard2, G.H. Wikfors3, S. Morton4, and T. Getchis5
Department of Marine Sciences, University of Connecticut, 1080 Shennecosset Road,
Groton, CT, 06340 USA
2
University of Hartford, Hillyer College, 200 Bloomfield Ave., West Hartford, CT 06117,
USA
3
NOAA, National Marine Fisheries Service, Northeast Fisheries Science Center, 212
Rogers Avenue, Milford, CT 06460, USA
4
NOAA, National Ocean Service, Marine Biotoxins Program, 219 Fort Johnson Rd.,
Charleston, SC 29412, USA
5
CT Sea Grant College Program, Department of Marine Sciences, University of
Connecticut, 1080 Shennecosset Road, Groton, CT 06340, USA
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
Biofouling ascidians are ubiquitous in coastal ecosystems and are among the main
colonizers of aquaculture gear. Our study tested the hypothesis that the transport,
removal, and transfer of fouling ascidian species by aquaculturists provide a mechanism
for concentration and distribution of harmful-algal cells to new areas. Wild-caught
specimens of common, biofouling ascidian species (Styela clava, Ciona intestinalis,
Molgula manhattensis, Bottryloides violaceus, Didemnum vexillum, and Botryllus
schlosseri) were exposed individually to cultured strains of co-occuring harmful algae
(Prorocentrum minimum, Alexandrium fundyense, Alexandrium monilatum, Karenia
brevis, Aureococcus anophagefferens, or Heterosigma akashiwo) at simulated bloom cell
densities of each HAB species. After feeding, ascidians were transferred to ultrafiltered
seawater. Immediately after exposure, and after 24 and 48 h in ultrafiltered seawater,
biodeposits were collected and observed microscopically for the presence of intact,
potentially-viable cells. Subsamples of biodeposits were transferred into culture tubes
with ultrafiltered seawater and monitored for algal growth during 8 weeks. Cells of all
HAB species were found to pass intact through the ascidian digestive system, remained
viable, and in many cases were capable of re-establishing populations at least 48 h
post-ingestion. The results of our study will inform industry and managers as to the
potential threat and ecological impact of spreading biofouling ascidians, and practices to
mitigate adverse impacts. Additionally, these management practices have been formally
incorporated into a new cost-share program developed to help shellfish producers
implement management practices to prevent the further spread of ascidians and
associated HAB species.
Page 184
THE INFLUENCE OF NUTRIENT AND LIGHT PERTURBATIONS ON
NATURAL PHYTOPLANKTON ASSEMBLAGES CONTAINING THE TOxIN
PRODUCING DIATOM PSEudO-nITZSCHIA
Jayme Smith1, Alyssa Gellene1, Paige Conne1l1, Victoria Campbell1, Erica Seubert1 and
David A. Caron1
University of Southern California, 3616 Trousdale Parkway, AHF 301, Los Angeles,
California 90089, USA
1
In March 2013, a multi-institutional ECOHAB field study was organized to characterize
the physical and biological parameters that are thought to influence bloom formation of
the toxin-producing diatom, Pseudo-nitszchia in the Southern California Bight. Pseudonitszchia blooms now occur with some regularity in the springtime in this region, an
observation that informed the timing of the field study. The principal hypothesis of
this study was that seed populations of Pseudo-nitszchia cells are advected from a
subsurface deep chlorophyll maximum to surface waters during upwelling events,
leading to a toxic event. Environmental parameters leading up to a bloom were
characterized, via ship based sampling and remote sensing.
Incubations 1 and 2 were conducted with seawater collected at the deep chlorophyll
maxima prior to obvious environmental indicators of a bloom event. A third incubation
was conducted with surface water collected during a minor Pseudo-nitszchia bloom
event as indicated by a regional shore based sampling effort. In each experiment,
communities were incubated with an array of nutrient and vitamin treatments, including
treatments with nitrate, ammonia, urea, and high and low light levels. Chlorophyll a
values indicate a general increase in biomass for incubations 1 and 2 with particular
growth observed in treatments containing nitrogen and vitamins. Incubation 3 showed
a general decrease in biomass except in the treatment containing vitamins. Overall
particulate domoic acid increased incrementally from below detectable levels for all
treatments in incubation 1. Domoic acid per cell in incubation 1 increased marginally
through the duration of the experiment with the increase occurring most obviously
in the high light urea treatment. A difference in domoic acid per cell in high light and
low light urea treatments was also observed. Domoic acid production remained below
detection in incubation 2, despite the stimulation of Pseudo-nitzschia cell grow in some
of the treatments. Domoic acid was down regulated in all treatments of incubation
3 despite measurable domoic acid per cell concentrations at the beginning of the
experiment.
ORAL PRESENTATIONS
POSTER
October 2013
S
In addition to field sampling efforts, three community incubation experiments were
conducted to test the effects of irradiance and nutrient availability on the growth
and toxin production of Pseudo-nitzschia cells presents in communities collected at
different time points. Each experiment was incubated for six days in ambient light and
temperature with a suite of samples collected, including samples for particulate domoic
acid, chlorophyll a, and preserved whole seawater at five discrete time points.
Page 185
UNPRECEDENTED mICROCySTIS BLOOM IN NORTHERN CALIFORNIA’S
BAY-DELTA ESTUARY AND POSSIBLE IMPACT ON ZOOPLANKTON
ABUNDANCE
William T Stringfellow1,3, Chelsea L Spier1, Jeremy Hanlon1,3, Mark Brunell1,2, Monica
Estiandan1,2, Teemu Koski4, and Juha Kääriä4
Ecological Engineering Research Program, School of Engineering & Computer Science,
University of the Pacific, 3601 Pacific Avenue, Stockton, CA 95211
2
Department of Biological Sciences, University of the Pacific, 3601 Pacific Avenue,
Stockton, CA 95211
3
Lawrence Berkeley National Laboratory Earth Sciences Division, 1 Cyclotron Road,
Berkeley, CA 94720
4
Turku University of Applied Sciences, Sepänkatu 1, Fin-20700 Turku, Finland
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
California’s Sacramento-San Joaquin Delta is an expansive fresh-water tidal estuary
that is part of the San Francisco Estuary, the largest estuary on the US Pacific coast.
Recently, there has been a collapse in the pelagic fish community in the San Francisco
Estuary, known as the pelagic organism decline (POD). Blooms of the cyanobacteria
Microcystis, which often produce the cyanotoxin microcystin, were first documented in
the Sacramento-San Joaquin Delta in 1999. Cyanotoxins have been suspected as one
contributing factor to POD. It has been proposed that microcystin could be contributing
to POD directly though poisoning of fish or indirectly by affecting zooplankton and
other food sources. In this study, we documented the spatial and temporal extent of
cyanobacteria blooms in the Southern Bay-Delta Estuary (south Delta), an area further
east and upstream in the San Francisco Estuary than previously studied. Copepod,
rotifer and total zooplankton abundance was examined in comparison to the spatial
distribution of microcystin. The environmental factors associated with cyanobacteria
blooms in the south Delta were investigated. Microcystis and other cyanobacteria were
common in the south Delta and a large, persistent Microcystis bloom was observed
in the summer of 2012, but not in 2009 or 2011. There was a strong relationship
between microcystin and cyanobacteria biomass (r2=0.74). In July and August of 2012,
microcystin concentrations were measured above California EPA recreational advisory
limits (0.8 µg/L) and World Health Organization drinking water limits (1.0 µg/L), with a
maximum observed concentration of 2.1 µg/L. In previous years, maximum observed
microcystin concentrations were below 0.1 µg/L in this study and in studies by others
(2004, 2005, 2007, and 2008). This bloom was associated significantly with lower density
and mass of zooplankton, including species which are important to a functioning foodweb in the Delta. We compared water quality, flow and climate conditions of 2012,
when the persistent bloom occurred, to 2009 and 2011. Significantly higher phosphorus
concentrations and temperatures occurred in 2012 corresponding to higher cyanotoxins
concentrations, and timing of Spring flows differed in 2012 compared 2009, both low
flow years, but cause and effect cannot yet be established. The implications of these
results in the context of the POD and climate change will be discussed.
Page 186
EPOxIDE HYDROLASES IN KAREnIA bREvIS: KEY ENZYMES IN
BREVETOxIN BIOSYNTHESIS
Pengfei Sun and Kathleen S. Rein
Department of Chemistry and Biochemistry, Florida International University, Miami, FL
33199
Epoxide hydrolases have been proposed to be key enzymes in the biosynthesis of
polyether ladder (PE) compounds such as the brevetoxins. These enzymes have the
potential to catalyze kinetically disfavored endo-tet cyclization reactions. As such,
they could have enourmous potential if applied toward semi-enzymatic syntheses. An
epoxide hydrolase which was identified in a Karenia brevis EST library has been cloned
and expressed for characterization. Kinetic parameters were evaluated using a variety
of epoxide substrates to assess substrate selectivity and enantioselectivity as well as its
potential to catalyze the critical endo-tet cyclization of epoxy alcohols.
ORAL PRESENTATIONS
POSTER
Page 187
S
October 2013
COMPARING THE DIEL VERTICAL MIGRATIONS OF SYMPATRIC
KARLOdInIum vEnEFICum (DINOPHYCEAE) AND CHATTOnELLA
SubSALSA (RAPHIDOPHYCEAE) IN LABORATORY COLUMNS
Charles L. Tilney1 and Mark E. Warner1
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
1
University of Delaware, 700 Pilottown Road, Lewes, DE 19958, USA
In a wide range of organisms, adaptation to the daily rise and fall of the sun has led to
diel rhythms in physiology and behavior. Diel rhythms, particularly those controlled
by molecular clocks (=endogenous circadian rhythms), are thought to enhance an
individual’s fitness by allowing organisms to utilize and preempt, cyclical changes
(temporal and spatial) in the biotic and abiotic environment. Factors such as light and
temperature vary predictably throughout the day, and in marine environments, factors
such as: light, temperature, nutrients, predators, and competitors can also be vertically
segregated in the water column. Many motile marine plankton have adapted to these
temporal and spatial gradients by performing diel vertical migrations (DVM) in addition
to exhibiting diel rhythms in photosynthesis and other metabolic processes. Here we
compare the diel rhythms in vertical migration and photophysiology in a mixotrophic
dinoflagellate (Karlodinium veneficum) and a mixotrophic raphidophyte (Chattonella
subsalsa) in a variety of artificial water column conditions. No comprehensive
assessment of DVM has yet been published for K. veneficum, and the comparison with a
sympatric raphidophyte may yield clues about these species’ relative reliance on DVM,
and on conditions conducive to affording a competitive advantage to one or the other.
Water columns were held in transparent acrylic columns (1.8m tall, holding 31L), each
illuminated from above by ramped-intensity LEDs. In-vivo chlorophyll fluorescence was
used to monitor the vertical distribution of cells, and active chlorophyll fluorescence was
used to assess PSII functionality with water column depth. Measurements were made
every 3h for 39h at 11 depths. In nutrient replete columns under 14:10 L:D cycles, both
species underwent diel vertical migrations, rising toward the surface during the daytime,
and receding to depth at night. During the day, C. subsalsa showed a single well-defined
near-surface peak, whereas K. veneficum migrated to a bi-modal distribution with 1
broad mid-depth peak, and 1 narrow surface peak. Migration to the surface or nearsurface in both species resulted in reductions in the maximum quantum yield of PSII (Fv/
Fm), likely due to photoprotective mechanisms or photodamage. K. veneficum exhibited
larger declines in Fv/Fm, that occurred earlier in the day than C. subsalsa, perhaps
explaining the formation of a secondary mid-depth biomass peak. Following several
days of L:D cycles, columns were shifted into continual darkness (D:D). Diel and depth
dependent periodicity of photochemistry noted under L:D cycles, was immediately
lost under continuous darkness in both species, wherein K. veneficum photochemistry
remained at the pre-dawn minimum level, while C. subsalsa photochemistry matched
night-time values. Furthermore, DVM in K. veneficum ceased in constant darkness, while
C. subsalsa continued DVM, suggesting that DVM is under endogenous circadian control
in C. subsalsa, but not in K. veneficum. The effect of continuous light and nutriclines will
be presented, with an assessment of the relative gains in carbon fixation from DVM in
both species.
Page 188
ENVIRONMENTAL FACTORS AFFECTING THE RELEASE OF DOPAMINE
AND OxIDANTS BY THE BLOOM-FORMING GREEN SEAWEED uLvARIA
ObSCuRA
Kathryn L. Van Alstyne, Katie J. Anderson, and Daniel H. van Hees
Shannon Point Marine Center, Western Washington University, 1900 Shannon Point
Road, Anacortes, WA, USA
Page 189
S
October 2013
ORAL PRESENTATIONS
POSTER
In Washington State, ulvoid macroalgae can form large blooms that have detrimental
effects on other organisms in the vicinity. A prominent component of these blooms
is the seaweed Ulvaria obscura, which grows in the lower intertidal and subtidal
zones. During spring tides, Ulvaria can be stranded on the shore during low tide were
it may be subjected to several stresses, including desiccation, high light levels, and
high temperatures. This alga can release a variety of compounds including dopamine
and oxidants, which are known to be toxic to other marine organisms. Using a series
of experiments that simulated conditions that might be encountered by low intertidal
algae, we quantified the release of dopamine and oxidants. Dopamine was released
when Ulvaria was desiccated then rehydrated, but was not released in response to high
water temperatures or different light conditions. It also was not released in response
to the presence of dopamine or exudates from other individuals. Oxidant release was
significantly lower in desiccated than undesiccated plants, regardless of the duration of
desiccation. This contrasts with results from another bloom-forming ulvoid alga, Ulva
lactuca, in which similar amounts of oxidants were released by undesiccated plants and
plants that had been desiccated for 1, 2, or 4 hours prior to rehydration. Our results
demonstrate that that the release of toxins by Ulvaria is affected by desiccation at low
tides in a species-specific manner.
THE DEVELOPMENT AND VALIDATION OF A MULTIPLEx QPCR ASSAY
FOR THE DETECTION OF TOxIN PRODUCING CYANOBACTERIA
Mark Van Asten1 Brett Neilan2 Jamal Al-Tebrineh2
Diagnostic Technology Pty Ltd, Suite 45, 7 Narabang Way, Belrose, NSW, 2085, Australia
School of Biotechnology and Biomolecular Sciences, University of New South Wales,
Sydney NSW 2052, Australia
1
2
We now have a good understanding of the genetic basis for toxin production by
a number of groups of microorganisms. The discovery of these toxin biosynthetic
pathways has enabled the development of genetic screening of environmental samples.
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Cyanotoxins pose a direct threat to public health. Molecular genetics underlying
cyanotoxin production in fresh and brackish water environments, but more specifically,
the toxin biosynthesis genes have been used to develop a multiplex quantitative PCR
assay for the detection and quantitation of micocystin, nodularin, cylindrospermopsin
and saxitoxin producing genes. Along with the copy number of the relevant cyanotoxin
biosynthesis gene an internal cyanobacteria-specific 16S rDNA control target is used as
a reference target. Validation and test data of the assay will be presented along with
details of the development of standards to each of the relevant target toxin genes to
which these sequences are quantitated against.
This multiplex quantitative PCR assay should become a very important addition to the
resources available to laboratories and authorities for better surveillance, detection,
prediction and monitoring of harmful algal blooms.
Page 190
IMMUNE FUNCTION IN RESCUED MANATEES ExPOSED TO BREVETOxINS
Catherine J. Walsha, Jennifer E. Yordya, Matthew B. A. Butawana, Courtney L. Bennetta,
Leanne Flewellingb, Ray Ballc, Martine De Witb, Katarina Englisha, Robert K. Bonded
Marine Immunology Program, Mote Marine Laboratory, Sarasota, FL, USA
St. Petersburg, FL, USA
c
Lowry Park Zoo, Tampa, FL, USA
d
Sirenia Project, Southeast Ecological Science Center, United States Geological Survey,
Gainesville, FL, USA
a
b
Page 191
S
October 2013
ORAL PRESENTATIONS
POSTER
The health of many Florida manatees (Trichechus manatus latirostris) is adversely
affected by exposure to blooms of the toxic dinoflagellate, Karenia brevis. K. brevis
blooms are common in manatee habitat areas off the southwestern coast of Florida and
produce a group of cyclic polyether toxins collectively referred to as red tide toxins, or
brevetoxins. Although the majority of manatees exposed to significant levels of red tide
toxins die, many manatees are also rescued as a result of sublethal exposure and are
successfully treated and returned to the wild. Sublethal exposure to brevetoxins may
potentially impact the manatee immune system. Lymphocyte proliferative responses
and a suite of immune function parameters in the plasma were used to evaluate effects
of brevetoxin exposure on manatee health in manatees rescued as a result of natural
exposure in their habitat. Healthy, unexposed manatees from Crystal River, FL, were
used for comparison of immune function parameters. Peripheral blood leukocytes
(PBL) isolated from whole blood of rescued manatees were stimulated with T-cell
mitogens, ConA and PHA. Significant decreases (P < 0.05) in lymphocyte proliferation
were observed in ConA or PHA stimulated lymphocytes collected from rescued animals
compared to unexposed animals. A suite of plasma parameters, including plasma
protein electrophoresis profiles, lysozyme activity, superoxide dismutase activity,
and reactive oxygen/nitrogen species, were also used to assess manatee health. A
significant correlation between plasma brevetoxin concentration and plasma ROS/RNS
concentrations was observed. No significant correlations were observed among other
immune function parameters measured in manatee plasma. To summarize, manatees
rescued from red tide toxin exposure demonstrate reduced lymphocyte proliferation
responses compared to healthy free-ranging manatees not exposed to brevetoxins.
A potential for increased oxidative stress in rescued manatees exists, with a positive
correlation between ROS/RNS activity and brevetoxin levels in the plasma. Sublethal
exposure to brevetoxins in the wild has the potential to impact immune function, and
thus overall health, in the Florida manatee.
MONITORING NUTRIENTS IN SODUS BAY NEW YORK
Samantha Weber1 and Gregory L. Boyer2
Drive, NY 13210
2
Drive, NY 13210
1
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
Located on the southern shore of Lake Ontario, Sodus Bay is an ecologically diverse
watershed that is also a popular tourist destination during the summer months. Bluegreen algal blooms are naturally occurring in Sodus Bay, but beginning in 2010 toxinproducing harmful algal blooms began occurring in concentrations high enough to cause
closures in some parts of the Bay. These blooms are likely to be driven by phosphorus
entering the Bay through the surrounding creeks and watershed. To help determine the
phosphorus loads to Sodus Bay from Sodus Creek East (the main creek entering Sodus
Bay), a continuous soluble reactive phosphorus monitoring system (WET Labs Cycle-P)
will be deployed just before the creek enters the Bay. This system should allow us to
determine the episodic phosphorus concentrations entering the bay from the creek.
This information will help us identify potential management options that may help
reduce the intensity and occurrence of these algal blooms in the future.
Page 192
ASSESSING THE STRENGTHS AND WEAKNESSES OF EACH FLORIDA
HAB REPORTING SYSTEM
Ben Whitenack1,2, Kate Kohler1, and Barbara Kirkpatrick1,3,4
University of Pennsylvania, Philadelphia, PA
3
Dept of Public Health Studies, Miller School of Medicine, Univ of Miami, Miami, FL
4
Rosenstiel School of Marine and Atmospheric Sciences, Univ of Miami, Miami, FL
1
2
The 2012-2013 Karenia brevis bloom impacting the west coast of Florida started in late
September, with approximately six months of human health impacts at the beach Also
in the Fall of 2013, he National Weather Service (NWS) instituted their new Harmful
Algal Bloom (HAB) reporting system, a “beach hazard statement”. The beach hazards
statements are pushed directly to various public instant weather alerting systems.
With this new alert system, there are now five HAB reporting systems in operation that
cover the Gulf coast of Florida; the Mote Marine Laboratory (MML) Beach Conditions
Reporting System, the Florida Fish and Wildlife Conservation Commission/Fish and
Wildlife Research Institute’s (FWC/FWRI) Red Tide status report, the National Oceanic
and Atmospheric Administration’s (NOAA) HAB Bulletin, and Florida Department of
Health in Sarasota County’s Healthy Beaches reports
Conclusions support a need for better evaluations from the user groups as well as
the intended stakeholders of each HAB reporting system in order to identify desired
prospective improvements to each system. Additionally, in order to increase the
marketability and usage of all of the systems, as all five systems collaborate with each
other, messaging of the benefits and weaknesses of each system needs more prominent
attention.
Page 193
S
October 2013
ORAL PRESENTATIONS
POSTER
A mixed method approach to assessing the Florida HAB reporting systems was used
to measure the intended usage of these five systems. A qualitative approach using
interviews of the administrators and creators of each of the systems was used to
understand the history of the system; the intended and actual audience; evaluations;
and desired prospective improvements. The quantitative method followed, comparing
Sarasota lifeguarded beach visitor counts at six beaches with the Karenia cell counts, the
usage numbers of the various HAB systems and temperature.
ROLE OF ROS SIGNALING IN THE DOMOIC ACID BIOSYNTHETIC
PATHWAY
April L. Woods1 and G. Jason Smith1
Moss Landing Marine Labs, 8272 Moss Landing Rd Moss Landing, CA 95039
1
POSTER
PRESENTATIONS
ORAL
PRESENTATIONS
This study investigates the role of cellular chemical signaling during initiation of Domoic
Acid (DA) biosynthesis in the HAB forming diatom Pseudo nitzschia. Environmental
parameters promoting DA production include silica limitation, source of nitrogen supply
and the availability of redox active metals (particularly iron and copper). This study
explores the role of nutrient stress and metal stress in the generation of Reactive Oxygen
Species (ROS), both biotically (intracellular) and abiotically (extracellular). When present
at levels higher than can be neutralized by cellular machinery, ROS cause damage to
cells; sometimes resulting in cell death. Conversely, low levels of ROS operate as a
normal part of healthy cell signaling, cueing the cell to generate a response.
An assay has been optimized to quantify the ROS generating potential of the cell free
environmental sample based on dithiothrietol oxidation. This assay is employed to
evaluate the correlation of ambient ROS activity with DA accumulation and potential
mitigation effects of DA on ROS active compounds. Profiles of ROS generating potential
in natural seawater, assessed from cruises taken off the Southern CA Bight and the
Monterey Peninsula conducted in Spring and Fall of 2013, respectively will be presented.
Variation in intracellular (pDA) vs extracellular (dDA) pools will be evaluated with respect
to patterns of ROS signaling observed. The ROS dependent fluorescence probe CarboxylH2DCFDA (CHDCFDA), evaluated by flow cytometry, is used to elucidate the magnitude
and dynamics of intracellular ROS signaling during stress challenge. Preliminary results
using fluorescence show that an immediate ROS burst is issued upon challenge with
a ROS generating reagent, but little ROS signal is seen if the culture is first allowed to
acclimate to the challenge before addition of the probe. Taken together, patterns of
response to extra and intracellular ROS challenges will help elucidate the role of ROS
signaling in regulation of DA biosynthesis by Pseudo-nitzschia.
Page 194
DETERMINATION OF PHYTOPLANKTON GROUPS USING TURNER
DESIGNS’ PHYTOFIND
Lawrence Younan
Turner Designs Turner Designs, Inc., 845 W. Maude Ave., Sunnyvale, CA 94085 USA
Page 195
S
October 2013
ORAL PRESENTATIONS
POSTER
Turner Designs’ PhytoFind is configured with multiple sensors, each with different
excitation/emission wavelengths designed to distinguish among phytoplankton groups
using group specific fluorescence characteristics. Preliminary lab data show positive
results for differentiating among phytoplankton groups, which is desirable for monitoring
efforts of most all aquatic habitats because it provides researchers with valuable data
that can be used for: predicting algal blooms, determining whether phytoplankton
communities have the potential to be harmful, monitoring algal stock used for feeding
aquaculture farms, detailed estimates of productivity and system turnover rates. The
problem with this type of measurement are interference factors, primarily turbidity and
dissolved organic material, that convolute the fluorescence response from various algal
groups making it difficult to accurately determine percent group contribution in a mixed
phytoplankton community. Empirically derived correction factors used for correcting
such interferences are disadvantageous because they don’t encompass the various
types of materials that exist, both as dissolved organics and suspended sediments.
The PhytoFind uses additional sensors configured to measure scattering and dissolved
organic materials, along with algal fluorescence, simultaneously. This results in the
generation of a correction factor per data point, increasing accuracy of measurement
greatly. The PhytoFind’s ability to accurately determine percent algal group contribution
and provide phytoplankton concentration estimates makes it a powerful research tool
for monitoring or characterizing algae in any type of system.
DEVELOPMENT OF CAPILLARY ELECTROPHORESIS AS PART OF A
MICRO-TOTAL ANALYTICAL SYSTEM FOR MICROCYSTINS
Bingxue Zheng, Bruce McCord and John P. Berry
POSTER
PRESENTATIONS
ORAL
ORAL
PRESENTATIONS
PRESENTATIONS
Department of Chemistry and Biochemistry, Florida International University, 11200 SW
8th Street, Miami, FL 33199, USA
The presence of “harmful algal blooms” (HAB) is a growing concern in aquatic
environments worldwide. Among HAB organisms, cyanobacteria are of special concern
because they have been reported worldwide to cause environmental and human health
problem through contamination of drinking water, and possibly bioaccumulation in
freshwater food-webs. Although several analytical approaches have been applied to
monitoring cyanobacterial toxins, conventional methods (e.g. ELISA, HPLC-MS) are
both costly and time-consuming such that analyses take weeks for field sampling and
subsequent lab analysis. The results obtained from the field sample, thus, reflect the
outdated data, but not the current status. Deployable micro total analytical system
(µTAS) for in situ detection of bloom toxins is emerging as a promising approach to
monitor these toxins. As part of on-going research, a deployable µTAS for real-time
and continuous detection of cyanotoxins, and specifically the microcystin (MC) family,
is currently being developed. Toward this end, capillary electrophoresis (CE) is a
particularly suitable method of analytical separation that can couple very small samples
(~1 nL) and rapid separations (≤2-5 min) to a wide range of selective and sensitive
detection techniques (e.g. UV, LIF, MS, ECD). Moreover, with respect to this on-going
research, CE is scalable to existing µTAS instrumentation, particularly including so-called
“lab-on-a-chip” (LOC) platforms. We will present current research into the optimization
of CE-based analysis of MCs, in relation to the future adaptation to previously developed
LOC-based µTAS, including investigation of analytical separation of MC variants (i.e.
modifications of buffer/solvent and pseudo-stationary phases) and subsequent detection
(e.g. highly sensitive laser-induced fluorescence versus UV detection).
Page 196
AUTHOR
INDEx
ORAL
ORAL
PRESENTATIONS
PRESENTATIONS
AUTHOR
INDEx S
a
Page 199
S
October 2013
AUTHOR INDEx
Blum, Patricia __________________________ 178
Abbott, Margaret _____________________ 45, 72
Boase, Nick ____________________________ 126
Abraham, Ann __________________________ 124
Bonde, Robert K. _______________________ 191
Abraham, Bill___________________________ 175 Borchert , Jerry __________________________ 46
Adams, Jillian __________________________ 110 Bossart, Gregory ________________________ 131
Adams, Rachel M ________________________ 78 Bottein, Marie-Yasmine____________________ 71 Allen, Allison L. __________________________ 86 Bourbonniere, Richard A___________________ 78
Al-Tebrineh, Jamal ______________________ 190
Bourdelais, Andrea ____________ 41, 89, 127, 137, 144, 146, 147, 154, 171, 173, 175
Anderson, Clarissa R. _________________ 31, 163 Bowers,
Holly
A.
__________________
73, 87, 115
Anderson, Donald M. ____________ 36, 39, 52, 59, 61, 65, 106, 110, 120, 140
Boyer, Gregory L ______________ 35, 78, 105, 107,
112, 113, 192
Anderson, Katie J. _______________________ 189 Boyes,
A.J.
_____________________________
128
Anderson, Paul E. _______________________ 145 Anglin, Scott B __________________________ 125
Brame, Julie ___________________________ 103
Archer, Stephen ________________________ 180
Brand, Larry________________________ 168, 170
Ardjmand, Ehsan_________________________ 64
Brandt, Ashley _________________________ 138
Asong, Jinkeng___________________________ 39
Bricelj, Monica _________________________ 182
Atkinson, Joseph _________________________ 35
Briggs, Travis _______________________ 119, 136
Austen, Melanie ________________________ 141
Bronk, Deborah __________________________ 47
Avila, Christian _________________________ 135
Brooks, Christopher _____________________ 103
Brosnahan, Michael L._________________ 36, 106
Brunell, Mark __________________________ 186
B
Bullard, S.G.____________________________ 184
Bachvaroff, Tsvetan R. ________________ 69, 158
Bumpus, Liz ____________________________ 76
Baden, Daniel G_________________ 127, 137, 144,
146, 147, 154, 171, 173, 175 Burks, Robert ______________________ 119, 136
Bailey, Trevor __________________________ 142
Burnell, Craig __________________________ 180
Baker, David_____________________________ 80
Burson, Amanda ________________________ 129
Ball, Ray ______________________________ 191
Butawan, Matthew B.A ___________________ 191
Banas, Neil S._________________________ 49, 65
Byrne, Jon ______________________________ 57
Barbe, Audrey _________________________ 182
Byrne, Margaret M ____________ 79, 88, 161, 162
Barbieri, Michelle ________________________ 71
Bargu, Sibel _____________________________ 77
C
Barron, Heather W. _______________________ 32
Campagna, Shawn R. _____________________ 78
Bartleson, Richard D. __________________ 32, 33
Campbell, Katrina ________________________ 39
Beck, Sheila ___________________________ 141
Campbell, Lisa _____________ 41, 48, 89, 118, 183
Becker, Richard __________________________ 35
Campbell, Victoria ______________________ 185
Beet, Andrew ______________________ 108, 153
Canfield, Dan __________________________ 119
Belisle, B. Shafer _________________________ 78
Carmichael, Wayne ______________________ 53
Bennett, Courtney L _________________ 131, 191
Caron, David A. ___________________ 51, 75, 185
Berry, Dianna L. __________________________ 56
Casamatta, Dale ________________________ 156
Berry, John P _________________ 34, 54, 116, 157, Cassell, Ryan ________________________ 90, 130
168, 196
Chao, Yi ________________________________ 51
Bianco, Colleen __________________________ 37
Charniga, Camia M. _____________________ 174
Bill, Brian D __________________________ 46, 65
Chauan, Vinita S. _______________________ 127
Blair, Stephen __________________________ 135
Chen, Fei ______________________________ 118
Bloomfield, Dan ________________________ 142
Chen, Wei _____________________________ 130
Choi, C.J. ______________________________ 139
Christman, Mary C _______________________ 91
Cichowska, Anna _______________________ 142
Clark, Quintin ___________________________ 76
Clement, Amy ___________________________ 95
Clemente, Jennifer _______________________ 76
Cocilova, Courtney ______________________ 131
Cody, Theresa ___________________________ 99
Collado-Vides, Ligia _____________________ 135
Connell, Laurie __________________________ 68
Connell, Paige ___________________________ 75
Connolly, Thomas ________________________ 49
Corcoran, Alina _________________________ 172
Cox, Annie _____________________________ 94
Coyne, Kathryn J _______________ 37, 70, 83, 102
Cray, Carolyn ____________________________ 32
Crespo, Bibiana G. ______________________ 110
Crump, Byron C. _________________________ 63
Currier, Robert _________________________ 149
AUTHOR INDEx
d
Daly, Kendra L. __________________________ 92
Darrow, Brian P. ________________________ 164
Davis, Edward ___________________ 98, 132, 133
Davis, Kristen____________________________ 49
de Bruijne, Wilco _______________________ 129
De Wit, Martine ________________________ 191
Dearth, Stephen P. _______________________ 78
Deeds, Jonathan R. _______________________ 38
Delwiche, Charles F. ______________________ 69
Depledge, Michael H_________________ 141, 142
Derner, Katherine ____________________ 98, 133
Dickey, Robert W________________________ 124
Dill, Brian D. ____________________________ 78
Dionysiou, Dionysios D ________________ 97, 177
Disney, Jane_____________________________ 52
Dixon, L. Kellie _______________ 47, 121, 134, 149, 164, 174
Doll, Cameron ___________________________ 37
Dou, Natalie ___________________________ 135
Doucette, Gregory J ________________ 39, 73, 87
Dreher, Theo W. _________________________ 66
Dupuy, Danielle ____________________ 119, 136
e
Eberhart, Bich-Thuy L._____________________ 46
Edwards, Christopher A ___________________ 31
Page 200
Effler, T. Chad ___________________________ 78
El Said, Kathleen R ______________________ 124
Eldred, K. C. _____________________________ 65
Elliott, Christopher T.______________________ 39
Elliott, Elizabeth A ___________ 127, 137, 144, 173
Ellsworth, Amanda ______________________ 138
Engene, Niclas ___________________________ 40
English, Katarina ________________________ 191
Erdner, Deana L. ____________________ 110, 139
Errera, Reagan M. _____________________ 41, 89
Estenik, John F. _________________________ 113
Estiandan, Monica ______________________ 186
f
Fahnenstiel, Gary L. ______________________ 64
Fahnenstiel, John A. ______________________ 64
Faris, Jeremy ___________________________ 108
Fernandez, Facundo M. ___________________ 57
Fine, Isaac ______________________________ 49
Fischer, Alexis D. ________________________ 140
Fleming, Lora E ______________ 50, 126, 141, 142, 153, 161 Flewelling, Leanne J __________ 52, 103, 117, 131,
150, 178, 191
Flores Quintana, Harold A____________ 52, 59, 72, 143
Foreman, Michael ________________________ 49
Fortuin, Anne __________________________ 129
Fowler, Nicholas ____________________ 144, 154
Frame, Elizabeth _________________________ 58
Frenzel, Hartmut _________________________ 51
Fridey, Kelly A. _________________________ 145
Fries, David _____________________________ 93
Fu, Feixue _____________________________ 117
Fulton, Rolland _____________________ 119, 136
Fuquay, Jennifer _________________________ 71
g
Gantar, Miroslav __________________ 34, 54, 157
Gao, Yonghui ____________________________ 74
Garcia, Ana C. ________________________ 59, 72
Garrett, Matthew J. ___________________ 47, 92
Garrison, B. ____________________________ 166
Gaze, William __________________________ 126
Geier, Susan_____________________________ 49
Gellene, Alyssa G. _________________ 51, 75, 185
Gerssen, Arjen _________________________ 129
Getchis, T. _____________________________ 184
Gibble, Corinne M. _______________________ 42
Gibbs, Patrick D. L. ____________________ 34, 54
Giddings, Sarah _________________________ 49
Gilmore, K. ____________________________ 128
Glibert, Patricia __________________________ 43
Gobler, Christopher J_______________ 44, 56, 159
Goebel, Nicole L _________________________ 31
Golding, Brian __________________________ 142
Goodman, Allan ________________ 127, 137, 146, 171, 173
Grandal, Meghan _______________________ 147
Granholm, April A. ______________________ 117
Grattan, Lynn M. ___________________ 45, 58, 72
Greenfield, Dianne I. __________________ 37, 148
Greengrove, Cheryl L. _________________ 65, 152
Gregson, Brian __________________________ 93
Gunasekera, Sarath P. _____________________ 40
Gutierrez, Danielle __________________ 156, 176
h
Ingraham, Helen L. _______________________ 32
J
Jacocks, Karl _______________ 127, 146, 171, 175
Jagus, Rosemary ____________________ 145, 158
Jaja-Chimedza, Asha ______________ 54, 116, 157
Janecek, Paul ___________________________ 93
Jayroe, David S. _________________________ 96
Jester, Edward L.E. ______________________ 124
Jiang, Wenjun ___________________________ 97
Jin, Di ________________________________ 153
Jochens, Ann E. _________________________ 55
Johnstone, J. A. __________________________ 65
Jones, Burton ___________________________ 51
Jones, Carrie ___________________________ 103
Jones, Christina __________________________ 57
Page 201
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i
AUTHOR INDEx
Hails, Alan _____________________________ 149
Haines, Andy __________________________ 142
Hajat, Shakoor _________________________ 142
Hall, Lauren _____________________________ 56
Hall, Nathan S.___________________________ 67
Hall, Sherwood _________________________ 150
Hallam, Steven __________________________ 94
Hambright, K. David _____________________ 151
Hanlon, Jeremy ________________________ 186
Hannach, Gabriela ______________________ 152
Hardison, D. Ransom______________________ 60
Harrington, Neil _________________________ 46
Harvey, Elizabeth L. ______________________ 62
Harvey, J. B. J. ___________________________ 73
Hatch, Marco B.A. _______________________ 94
Hattenrath-Lehmann, T.K.__________________ 44
Hauser, Loren J. _________________________ 78
Hay, L. R. _______________________________ 65
Hayashi, Kendra__________________ 51, 115, 163
Heil, Cynthia ____________________________ 47
Hendrickson, John __________________ 119, 136
Henrichs, Darren W. ______________________ 48
Henry, Chuck ___________________________ 76
Henry, Michael _________________________ 178
Henschen, Karen _______________________ 103
Hetland, Robert D. ___________________ 48, 118
Hickerson, Emma L._______________________ 81
Hickey, Barbara __________________________ 49
Higginbotham, Tom ______________________ 76
Hiolski, Emma ___________________________ 58
Hitchcock, Gary _________________ 47, 50, 92, 95, 101, 161
Hoagland, Porter ____________ 50, 101, 108, 111, 153, 161 Hogan, Sarah __________________________ 148
Hogue, Tanya __________________________ 154
Holland, William C. _______________________ 81
Holmes, Sarah M. _______________________ 155
Holohan, B.A ___________________________ 184
Hooe-Rollman, Jennifer I. __________________ 72
Hoogenboom, Ron ______________________ 129
Hossain, Sk Md Sazzad ___________________ 177
Hounshell, Alexandria G. _________________ 174
Howard, Meredith D.A.____________________ 51
Hoyer, Mark ___________________________ 119
Huang, I-Shuo __________________________ 156
Hubbard, Katherine A. _____________ 52, 93, 103
Hudnell, H. Kenneth ______________________ 53
Hudson, Derrick S. ______________________ 101
Huisman, Jef ___________________________ 129
Hutchins, David A. ______________________ 117
Jones, Grant D. _________________________ 158
Jones, William J. ________________________ 148
K
AUTHOR INDEx
Kääriä, Juha ___________________________ 186
Kachel, Nancy ___________________________ 49
Kang, Yoonja ________________________ 56, 159
Kashinsky, Liz ___________________________ 71
Kaufman, Angie _________________________ 71
Kavanaugh, Karen ________________ 98, 132, 133
Keafer, Bruce A.___________________ 39, 61, 110
Kendrick, Preston ____________________ 58, 160
Keppler, Chuck _________________________ 148
Kessel, Anthony ________________________ 142
Khidekel, Roman _______________________ 113
Kibler, Steven R. _________________________ 81
Kiene, Ronald P. ________________________ 143
Killberg-Thorsen, Lynn ____________________ 47
Kirkpatrick, Barbara _____________ 50, 55, 79, 88, 95, 101, 111, 153, 161, 162, 193
Kirkpatrick, Gary _______________ 47, 50, 95, 101, 121, 134, 149, 161, 164
Kleindinst, Judith L. _______________________ 52
Knight, Christopher _____________________ 103
Koch, Florian ________________________ 56, 159
Kohler, Kate ___________________ 50, 79, 88, 95, 111, 161, 162, 193, Koski, Teemu __________________________ 186
Krausfeldt, Lauren E. ____________________ 107
Kubanek, Julia __________________________ 57
Kudela, Raphael M ______________ 31, 42, 49, 51, 87, 115, 163, 179, Kulis, David M. _________________________ 120
l
Landsberg, Jan H. _____________________ 91, 99
Langlois, Gregg _________________________ 150
Lasi, Margaret ___________________________ 56
Launay, Susan ______________________ 121, 174
Lazensky, Rebecca ______________________ 108
LeCleir, Gary R. _________________________ 107
Ledreux, Aurelie _________________________ 71
Lefebvre, Kathi ______________________ 58, 160
Leliaert, Frederik _______________________ 135
Lembke, Chad __________________________ 149
Lenes, Jason M.__________________ 47, 134, 164
Page 202
Lessard, Evelyn J. ____________________ 49, 167
Leynse, Alexander K. ____________________ 165
Li, Zhou ________________________________ 78
Li, Zongchao ___________________________ 166
Liefer, Justin D. _______________________ 59, 72
Linton, Brian ____________________________ 91
Lirman, Diego __________________________ 135
Litaker, R. Wayne _____________________ 60, 81
Littan, Charles ___________________________ 71
Liu, Shunmei ___________________________ 168
Loeffler, Christopher R. __________________ 100
Logsdon, Jenna L. _______________________ 177
Lovko, Vincent J. ______________ 50, 95, 101, 121
Lubetkin, Susan C. ___________________ 49, 167
Luther, Mark E. __________________________ 92
Lydon, Christina ________________________ 168
m
MacCready, Parker _______________________ 49
MacLeod, B. ___________________________ 128
Main, Christopher R. _________________ 37, 102
Makarewicz, Joseph ______________________ 35
Mantua, N. J. ____________________________ 65
Marcinek, David _____________________ 58, 160
Marin, Roman ________________________ 39, 87
Markley, Laura R. _______________________ 169
Marriott, Ian ___________________________ 127
Martin, Ned ___________________________ 175
Martin, Robbie M. ______________________ 107
Marzolf, Erich ______________________ 119, 136
Masson, Diane __________________________ 49
Masura, J. E. ____________________________ 65
Matthijs, Hans C.P. ______________________ 129
May, Amanda L. _________________________ 78
Maze, Grace ___________________________ 170
McCabe, Ryan ___________________________ 49
McCall, Jennifer ________________ 146, 171, 175
McCord, Bruce _________________________ 196
McGillicuddy, Dennis J. _________________ 52, 61
McInnis, Katherine B. _____________________ 32
McLaughlin, Karen _______________________ 51
McLean, Timothy I. ___________________ 96, 125
Medvecky, Rebecca _____________________ 178
Meek, Megan E. ________________________ 172
Menden-Deuer, Susanne __________________ 62
Mendez, Hector _________________________ 76
Mengel, Michael J. _______________________ 51
Meyer, Kevin A. _______________________ 47, 63
Mihuc, Timothy __________________________ 35
Mikulski, Christina M. _______________ 39, 73, 87
Milbrandt, Eric C. ________________________ 33
Miller, Jan D. ____________________________ 56
Millie, David F. __________________________ 64
Milton, Sarah __________________________ 131
Minotti, Patricia J. ______________________ 174
Moeller, Peter __________________________ 156
Moisander, Pia H. _______________________ 155
Moore, Leslie____________________________ 46
Moore, Stephanie K. ___________________ 65, 94
Moran, N. _____________________________ 128
Morey, Jeanine S. ____________________ 82, 145
Morris, George _________________________ 141
Morris, J. Glenn ____________________ 45, 58, 72
Morton, Steve _______________________ 52, 184
Mueller, Karl ____________________________ 94
Mulholland, Margaret R._______________ 47, 164
p
n
r
Neilan, Brett ___________________________ 190
Neill, Nina E. ___________________________ 173
Nezlin, Nikolay___________________________ 51
Nilsson, William _________________________ 94
Nissanka, Ari ___________________ 121, 134, 174
Niven, Susan ____________________ 41, 171, 175
Nunn, Brook ____________________________ 57
o
Qin, Boqiang ____________________________ 67
Radan, Regina __________________________ 179
Radke, Elizabeth G. ____________________ 45, 72
Ralston, David K. _________________________ 36
Ramsdell, John __________________________ 71
Rauschenberg, Carlton ___________________ 180
Redshaw, Clare _________________________ 126
Reich, Andrew _______________ 50, 108, 153, 161
Rein, Kathleen S. _________________ 90, 130, 187
Reis, Stefan ____________________________ 141
Rhodes, Linda ___________________________ 94
Richards, R. Peter ________________________ 80
Richardson, Bill __________________ 47, 109, 181
Richlen, Mindy L._____________________ 59, 110
Roberts, Sparkle M. ________________ 45, 58, 72
Robertson, Alison _______________ 45, 52, 59, 72,
100, 143, Robertson, George ____________________ 51, 75
Rodriguez, Dení ________________________ 135
Rolton, Anne ___________________________ 182
Rosa, M. ______________________________ 184
Roy, Jessie ______________________________ 57
Rudge, Katrin __________________________ 111
Ryan, Darcie ________________________ 89, 183
Ryan, John P. _________________________ 73, 87 Page 203
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October 2013
Q
AUTHOR INDEx
O’Dea, Sheila N. _____________________ 52, 103
O’Mara, Sean P. ________________________ 176
O’Neil, Judy M. _______________________ 47, 63
O’Neill, Charles __________________________ 35
O’Shea, Kevin E. _____________________ 97, 177
Oesch, Carl ____________________________ 113
Ogle, Marcus T. _________________________ 118
Olascoaga, Josefina _____________________ 170
Olsen, David A. _________________________ 100
Olson, Robert J. __________________________ 36
Osborne, Nick __________________________ 142
Otten, Timothy G. _____________________ 66, 67
Owen, Jenna L. _________________________ 104
Owen, Daniel P. ________________________ 104
Owen, Kevin C. _________________________ 104
Paerl, Hans W. ________________________ 66, 67
Palubok, Val ___________________________ 121
Paolisso, Michael_________________________ 74
Parsons, Michael L. _____________ 138, 165, 169
Paul, Matthew R. _______________________ 145
Paul, Valerie J. ___________________________ 40
Perri, Katherine A. ______________________ 105
Phillips, Jennifer _________________________ 68
Pierce, Richard _____________________ 178, 182
Pitz, Kathleen __________________________ 106
Place, Allen R. ________________ 69, 74, 145, 158
Plakas, Steven M. _______________________ 124
Pokrzywinski, Kaytee L. ___________________ 70
Poulson-Ellestad, Kelsey ___________________ 57
Pound, Helena L. _______________________ 107
Procise, Leo _____________________________ 47
AUTHOR INDEx
S
Sabel, Clive ____________________________ 142
Salathé, E. P. ____________________________ 65
Sandifer, Paul A. _________________________ 86
Sarran, Christophe ______________________ 142
Savage, Marci L. ________________________ 112
Savage, Thomas J. _______________________ 115
Sayers, Michael J. ________________________ 64
Scheller, Karen _________________________ 161
Schmale, Michael C. ______________________ 34
Schmidt, Justine R. __________________ 107, 113
Schneck-Scott, Adria ______________________ 98
Schneider, Sabrina ______________________ 135
Schnetzer, Astrid _________________________ 75
Scholin, Christopher A____________ 39, 73, 87, 94
Scott, Paula ____________________________ 103
Sellner, Kevin G. _________________________ 74
Senqupta, Ashmita _______________________ 51
Seubert, Erica L. __________________ 51, 75, 185
Shankar, Sugandha ______________________ 114
Shaskus, Mylynda _______________________ 113
Shawvner-Karnitz, A._____________________ 128
Shea, Damian ___________________________ 60
Shipley, Dianne __________________________ 76
Shoesmith, Zoe __________________________ 95
Shuchman, Robert A. _____________________ 64
Shumway, Sandra E. _________________ 182, 184
Siedlecki, Samantha ______________________ 49
Silby, Mark W. __________________________ 155
Sipler, Rachel ____________________________ 47
Sirois, Alison ____________________________ 52
Smith, Don______________________________ 58
Smith, Emily A. __________________________ 77
Smith, G. Jason ______________ 87, 115, 163, 194
Smith, Jayme ________________________ 75, 185
Smith, Juliette L. _____________________ 39, 120
Smith, T.________________________________ 59
Sosik, Heidi M.___________________________ 36
Soudant, Philippe _______________________ 182
Southard, Greg _________________________ 176
Spier, Chelsea L. ________________________ 186
Stark, Kimberle _________________________ 152
Steele, Dalton D. ________________________ 116
Steffen, Morgan M. ______________________ 78
Steidinger, Karen A.____________________ 27, 91
Stein, J. E. ______________________________ 65
Page 204
Steur, Kees ____________________________ 129
Stomp, Maayke ________________________ 129
Stringfellow, William T. ___________________ 186
Strom, Mark ____________________________ 94
Studts, Jamie L. _______________ 79, 88, 161, 162
Stumpf, Richard P. ______________ 80, 86, 91, 101, 119, 136
Sultan, Mohammed ______________________ 35
Sun, Pengfei ___________________________ 187
Sunda, William G. ________________________ 60
Sutula, Martha __________________________ 51
Sweeney, Pamela _______________________ 135
t
Talens, Renee __________________________ 129
Tang, Ying Zhong _________________________ 56
Tatters, Avery O. ________________________ 117
Taylor, Alison __________________________ 147
Taylor, Tim ____________________________ 141
Tester, Patricia A.______________________ 60, 81
Thomas, Maura. A. ___________________ 61, 114
Thompson, Alysha ______________ 146, 171, 175
Thompson, Mark A. ______________________ 33
Thomson, Richard ________________________ 49
Thyng, Kristen M. _______________________ 118
Tiedeken, Jessica ________________________ 71
Tilney, Charles L. _____________________ 83, 188
Tomas, Carmelo ________________________ 144
Tomlinson, Michelle C. ____________ 80, 119, 136
Tong, Mengmeng _______________________ 120
Townsend, David W. __________________ 61, 114
Trainer, Vera L. ____________________ 46, 65, 94
Tustison, Jacob __________________________ 91
u
Ullmann, Steve __________________ 50, 153, 166
Urizar, Cristina _____________________ 132, 133
v
Van Alstyne, Kathryn L. __________________ 189
Van Asten, Mark _______________________ 190
Van Dolah, Frances M _____________ 82, 145, 150,
172
van Hees, Daniel H. _____________________ 189
van Scheppingen, Yvonne ________________ 129
Vandersea, Mark W. ______________________ 81
VanDolah, Elizabeth ______________________ 74
VerBerkmoes, Nathan C. __________________ 78
Viant, Mark _____________________________ 57
Vignier, Julien __________________________ 182
Visser, Petra M. ________________________ 129
Volety, Aswani _________________________ 182
Vreeland, Jennifer M ____________ 121, 134, 174, W
Wallace, Ryan B. _________________________ 44
Walsh, Breanne __________________________ 47
Walsh, Catherine J. __________________ 131, 191
Walsh, John J. _______________________ 47, 164
Walter, John ____________________________ 91
Walters, Linda ___________________________ 56
Walton, Katherine ________________________ 34
Wang, Zhihong __________________________ 71
Warner, Mark E. __________________ 70, 83, 188
Watkins, Sharon ________________________ 153
Watson, Sue B. _______________________ 35, 78
Watzin, Mary____________________________ 35
Weber, Samantha _______________________ 192
Weckman, Gary R. _______________________ 64
Weidert, Chase __________________________ 77
Weisberg, Robert ________________________ 47
White, Mathew ____________________ 126, 141
Whitenack, Ben ____________________ 162, 193
Wikfors, G.H. ___________________________ 184
Wilhelm, Steven W. _______________ 35, 78, 107
Williams, Ernest ______________________ 69, 74
Wilson, Patrick __________________________ 99
Wolfe, Steven H. _________________________ 55
Wolny, Jennifer _________________________ 103
Woods, April L. _________________________ 194
Wren, Jeff _____________________________ 103
Wynne, Timothy T. _______________________ 80
x
Xu, Hai _________________________________ 67
y
Yordy, Jennifer E. ___________________ 131, 191
York, Bob _____________________________ 144
Younan, Lawrence _______________________ 195
Young, William A., II ______________________ 64
z
Zamor, Richard M. ______________________ 151
Zaworski, Lori J. ________________________ 174
Zhang, Xiaoqian ________________________ 118
Zhang, Y. _______________________________ 73
Zheng, Bingxue _________________________ 196, Zhu, Guangwei __________________________ 67
Zimba, Paul V. ______________________ 156, 176
AUTHOR INDEx
Page 205
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October 2013
PARTICIPANT
LIST
ORAL
ORAL
PRESENTATIONS
PRESENTATIONS
PARTICIPANT
LIST S
ann abraham
P.O. Box 158, 1 Iberville Drive
ann.abraham@fda.hhs.gov
Clarissa anderson
University of California Santa Cruz
1156 High Street
Santa Cruz, CA 95064
clrander@ucsc.edu
don anderson
Woods Hole Oceanographic Institution
MS #32
Woods Hole, MA 02543
danderson@whoi.edu
Scott anglin
The University of Southern Mississippi
118 College Drive, #5018
Hattiesburg, MS 39406
scott.anglin@eagles.usm.edu
dan ayres
Washington Department of Fish and
Wildlife
119 Brady Loop Road East
Montesano, WA 98563
daniel.ayres@dfw.wa.gov
heather Barron, dvm
CROW, Inc.
PO Box 150
Sanibel, FL 33957
heather@crowclinic.org
patricia Blum
1600 Ken Thompson Parkway
Sarasota, FL 34236
pcblum@mote.org
Jerry Borchert
Washington State Department of Health
P.O. Box 47824
Olympia, WA 98504-7824
jerry.borchert@doh.wa.gov
andrea Bourdelais
University of North Carolina Wilmington
Center for Marine Science
5600 Marvin K. Moss Lane
Wilmington, NC 28409
bourdelaisa@uncw.edu
holly Bowers
Monterey Bay Aquarium Research Institute
7700 Sandholdt Road
Moss Landing, CA 95039
hbowers@mbari.org
greg Boyer
State University of New York - ESF
1 Forestry Drive
Syracuse, NY 13210
glboyer@esf.edu
Page 209
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October 2013
John Berry
Florida International University
3000 NE 151st Street
North Miami, FL 33181
berryj@fiu.edu
PARTICIPANT LIST
lorraine C. Backer
Centers for Disease Control
National Center for Environmental Health
4770 Buford Highway NE
MS F-60
Chamblee, GA 30341
lfb9@cdc.gov
richard Bartleson
Sanibel-Captiva Conservation Foundation
Marine Laboratory
900 A Tarpon Bay Road
Sanibel, FL 33957
rbartleson@sccf.org
anamari Boyes
Manatee County Water Treatment Plant
QC Lab
17915 Waterline Road
Bradenton, FL 34212
anamari.boyes@mymanatee.org
ashley Brandt
Florida Gulf Coast University
10501 FGCU Blvd. South
Fort Myers, FL 33965
albrandt@eagle.fgcu.edu
michael Brosnahan
Redfield 3-30, MS 32
mbrosnahan@whoi.edu
PARTICIPANT LIST
amanda Burson
University of Amsterdam
904 Science Park
Amsterdam, the Netherlands
1098 XH
a.m.o.burson@uva.nl
meridith Byrd
Texas Parks and Wildlife
2805 North Navarro, Suite 600A
Victoria, TX 77901
meridith.byrd@tpwd.texas.gov
margaret Byrne
University of Miami
Department of Public Health Services
Miller School of Medicine
1120 NW 14th Street, Suite 912
Miami, FL 33136
mbyrne2@med.miami.edu
lisa Campbell
Texas A&M University
3146 TAMU
College Station, TX 77843
lisacampbell@tamu.edu
Page 210
Wayne Carmichael
Department of Biological Sciences
Wright State University
Dayton, OH 45435
wayne.carmichael@wright.edu
ryan Cassell
875 NW 2nd Street
Miami, FL 33128
rcass004@fiu.edu
Camia Charniga
Sarasota, FL 34236
camia@mote.org
Wei Chen
11200 SW 8th Street
Miami, FL 33199
wchen020@fiu.edu
mary Christman
MCC Statistical Consulting
2219 NW 23rd Terrace
Gainesville, FL 32605
marycchristman@gmail.com
Courtney Cocilova
Florida Atlantic University
777 Glades Road
Boca Raton, FL 33431
ccocilov@fau.edu
Jaime Cook
Florida Department of Health
in Collier County
Environmental Health and Engineering
3339 Tamiami Trail East, Suite 145
Naples, FL 34112
jaime_cook@doh.state.fl.us
alina Corcoran
Florida Fish and Wildlife Conservation
Commission
Fish and Wildlife Research Institute
100 8th Avenue SE
St Petersburg, FL 33701
alina.corcoran@MyFWC.com
Kathryn Coyne
University of Delaware
School of Marine Science and Policy
700 Pilottown Road
Lewes, DE 19958
kcoyne@udel.edu
edward davis
NOAA National Ocean Service
1305 East West Highway
Silver Spring, MD 20910
Edward.Davis@noaa.gov
Jonathan deeds
5100 Paint Branch Parkway
College Park, MD 20740
jonathan.deeds@fda.hhs.gov
l. Kellie dixon
Sarasota, FL 34236
lkdixon@mote.org
Quay dortch
NOAA NCOS
Building IV, Room 8218
quay.dortch@noaa.gov
danielle dupuy
NOAA/NCCOS & CSS Dynamic
10301 Democracy Lane, Suite 300
Fairfax, VA 22030
danielle.dupuy@noaa.gov
elizabeth elliott
elliotte@uncw.edu
amanda ellsworth
10501 FGCU Blvd South
aellsworth@fgcu.edu
niclas engene
Smithsonian Marine Station at Ft. Pierce
701 Seaway Drive
Fort Pierce, FL 34949
engenen@si.edu
ivory engstrom
McLane Research Labs
121 Bernard Saint Jean Drive
East Flamouth, MA 02536
Iengstrom@mclanelabs.com
Page 211
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October 2013
greg doucette
219 Fort Johnson Road
Charleston, SC 29412
greg.doucette@noaa.gov
PARTICIPANT LIST
Katherine derner
672 Independence Parkway
Chesapeake, VA 23320
katie.derner@noaa.gov
natalie doucette
11200 SW 8th Street, OE 167
Miami, FL 33199
ndou1@fiu.edu
deana erdner
University of Texas Marine Science
Institute
750 Channel View Drive
Port Aransas, TX 78373
derdner@utexas.edu
reagan errera
3146 TAMU
rmerrera@tamu.edu
alexis fischer
266 Woods Hole Road
Redfield 332 (MS #32)
afischer@whoi.edu
PARTICIPANT LIST
timothy fitzpatrick
Saigene Biotech
1660 Race Street
Denver, CO 80206
tfitzpatrick@saigene.com
lora fleming
European Centre for Environment and
Human Health
University of Exeter Medical School
Truro Cornwall, United Kingdom
TR1 3HD
lora.fleming@pcmd.ac.uk
leanne flewelling
Commission
100 8th Ave SE
leanne.flewelling@myfwc.com
Page 212
harold flores Quintana
harold.floresquintana@fda.hhs.gov
nicholas fowler
nlf7589@uncw.edu
hugo freudenthal
Long Island University (Retired)
1524 Fairway Drive
Dunedin, FL 34698
hfreudenthal@knology.net
anita freudenthal
Nassau County Dept. of Health (Retired)
1524 Fairway Dr.
Dunedin, FL 34698
hfreudenthal@knology.com
Kelly fridey
College of Charleston
frideyka@g.cofc.edu
matthew garrett
Commission
100 8th Avenue SE
matt.garrett@ myfwc.com
Corinne gibble
University of California, Santa Cruz
217 Highland Ave. Apt. 1
cgibble@ucsc.edu
pat glibert
University of Maryland Center for
Environmental Science
P.O. Box 775
Cambridge, MD 21613
glibert@umces.edu
Christopher gobler
School of Marine and Atmospheric
Sciences
Stony Brook University
Stony Brook, NY 11794
christopher.gobler@stonybrook.edu
allan goodman
goodmana@uncw.edu
meghan grandal
601 S. College Road
mkg7188@uncw.edu
dianne greenfield
University of South Carolina
Hollings Marine Laboratory
dgreenfield@belle.baruch.sc.edu
Brian gregson
Spyglass Technologies, Inc.
101 16th Avenue South, Suite 4A
St. Petersburg, FL 33701
brian.gregson@spyglasswater.com
K. david hambright
Department of Biology
University of Oklahoma
730 Van Vleet Oval
304 Sutton Hall
Norman, OK 73019
dhambright@ou.edu
gabriela hannach
King County Environmental Lab
322 West Ewing Street
Seattle, WA 98119
gabriela.hannach@kingcounty.gov
Clara hard
Washington State Dept of Health
P.O. Box 47824
Olympia, WA 98504
clara.hard@doh.wa.gov
Samantha harlow
Sarasota, FL 34236
sharlow@mote.org
neil harrington
Jamestown S’Klallam Tribe
1033 Old Blyn Hwy.
Sequim, WA 98282
nharrington@jamestowntribe.org
Page 213
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October 2013
alan hails
Sarasota, FL 34236
ahails@mote.org
PARTICIPANT LIST
lynn grattan
University of Maryland
School of Medicine
110 S. Paca Street
Baltimore, MD 21201
lgrattan@som.umaryland.edu
andrew griffith
Stony Brook, NY 11794
awgriffit@gmail.com
PARTICIPANT LIST
marco hatch
Northwest Indian College
2522 Kwina Road
Bellingham, WA 98226
marcoh@nwic.edu
porter hoagland
MS#41, Marine Policy Center
phoagland@whoi.edu
Cynthia heil
Bigelow Laboratory for Ocean Sciences
60 Bigelow Dr., P.O. Box 380
East Boothbay, ME 04544
cheil@bigelow.org
tanya hogue
hoguet@uncw.edu
John hendrickson
St. Johns River Water Management District
4049 Reid Street
Palatka, FL 32177
jhendrickson@sjrwmd.com
Sarah holmes
University of Massachusetts Dartmouth
1399 Phillips Rd
New Bedford, MA 02745
sholmes1@umassd.edu
darren henrichs
3146 TAMU, Eller O&M Bldg
dhenrichs@tamu.edu
alexandria hounshell
Sarasota, FL 34236
ahounshell@mote.org
michael henry
1600 Ken Thompson Pky
Sarasota, FL 34236
mhenry@mote.org
meredith howard
Southern California Coastal Water
Reserach Project
3535 Harbor Blvd. Suite 110
Costa Mesa, CA 92626
mhoward@sccwrp.org
Barbara hickey
University of Washington
School of Oceanography
Box 355351
Seattle, WA 98022
bhickey@u.washington.edu
i-Shuo huang
Texas A&M University-Corpus Christi,
Center for Coastal Studies
7350 McArdle Rd.
ihuang@islander.tamucc.edu
gary hitchcock
University of Miami
4600 Rickenbacker Causeway
Miami, FL 33149
g.hitchcock@miami.edu
Kate hubbard
Commission
100 8th Avenue SE
katherine.hubbard@MyFWC.com
Page 214
h. Kenneth hudnell
Medora Corp. & UNC-Chapel Hill
105 Serrano Way
Chapel Hill, NC 27517
ken.hudnell@medoraco.com
derrick hudson
Sarasota, FL 34236
derrickshudson@gmail.com
asha Jaja-Chimedza
3000 N.E. 151st Street, MSB 332
ajaja001@fiu.edu
david Jayroe
University of Southen Mississippi
118 College Drive, #5018
Hattiesburg, MS 39406
david.jayroe@eagles.usm.edu
Wenjun Jiang
11200 SW 8th Street
Miami, FL 33199
wjian001@fiu.edu
heather Johnson
U.S. Geological Survey
6520 Mercantile Way
Suite 5
Lansing, MI 48911
hejohnson@usgs.gov
yoonja Kang
239 Montauk Highway.
Southampton, NY 11968
yoonja.kang@stonybrook.edu
david Karlen
Environmental Protection Commission
of Hillsborough County
P.O. Box 1110
Tampa, FL 33601
karlen@epchc.org
Karen Kavanaugh
1305 East-West Highway
Silver Springs, MD 20910
karen.kavanaugh@noaa.gov
preston Kendrick
NOAA Fisheries
2725 Montlake Blvd East
Seattle, WA 98112
preston.kendrick@noaa.gov
Barb Kirkpatrick
Sarasota, FL 34236
bkirkpat@mote.org
Page 215
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October 2013
Sonia Joseph Joshi
NOAA Center of Excellence for
Great Lakes and Human Health
4840 South State Road
Ann Arbor, MI 48108
sonia.joseph@noaa.gov
PARTICIPANT LIST
ann Jochens
GCOOS
3146 TAMU
ajochens@tamu.edu
grant Jones
University of Maryland
Center for Environmental Sciences
701 East Pratt Street
Baltimore, MD 21202
gdjones@umaryland.edu
gary Kirkpatrick
Sarasota, FL 34236
gkirkpat@mote.org
Judy Kleindinst
MS #32, Redfield 332
jkleindinst@whoi.edu
florian Koch
School of Marine and Atmospheric
Sciences
239 Montauk Highway
Southampton, NY 11968
floriankoch@gmail.com
Kate Kohler
Sarasota, FL 34236
kkohler@mote.org
PARTICIPANT LIST
Julia Kubanek
Georgia Institute of Technology
School of Biology, 310 Ferst Drive
Atlanta, GA 30332
julia.kubanek@biology.gatech.edu
raphael Kudela
Ocean Sciences Department
1156 High Street
kudela@ucsc.edu
Jan landsberg
Commission
100 8th Ave SE
jan.landsberg@myfwc.com
Page 216
Brian lapointe
Harbor Branch Oceanographic Institute
777 Glades Road
blapoin1@hboi.fau.edu
Susan launay
Sarasota, FL 34236
susans@mote.org
Stephanie lear
Mote Marine Lab
Sarasota, FL 34236
lears@mote.org
Kathi lefebvre
NOAA Fisheries
Northwest Fisheries Science Center
2725 Montlake Boulevard East
Seattle, WA 98112
kathi.lefebvre@noaa.gov
Jason lenes
University of South Florida
140 7th Avenue South
lenes@marine.usf.edu
alex leynse
akleynse@eagle.fgcu.edu
zangchao (Cathy) li
University of Miami
School of Communication
5150 Brunson Drive
z.li13@umiami.edu
Justin liefer
justin.liefer@fda.hhs.gov
rob magnien
DOC/NOAA/NOS/CSCOR
rob.magnien@noaa.gov
Wayne litaker
NOAA
101 Pivers Island Dr.
Beaufort, NC 28516
wayne.litaker@noaa.gov
Christopher main
700 Pilottown Road
Lewes, DE 19958
cmain@udel.edu
Christopher loeffler
christopher.loeffler@fda.hhs.gov
laura markley
Commission
100 8th Ave SE
laura.markley@myfwc.com
Keith loftin
U.S. Geological Survey
4821 Quail Crest Place
Lawrence, KS 66049
kloftin@usgs.gov
vince lovko
Mote Marine Laboratories
1600 Ken Thompson Pkwy
Sarasota, Fl 34236
vlovko@mote.org
Christina lydon
3000 NE 151st Street
chlydo@fiu.edu
dennis mcgillicuddy
Bigelow 209 - MS 11
dmcgillicuddy@whoi.edu
megan meek
Graduate Program in Marine Biology,
College of Charleston
meekme@g.cofc.edu
Page 217
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October 2013
Jennifer mcCall
mccalljr@uncw.edu
PARTICIPANT LIST
Susan lubetkin
University of Washington
Box 357940
Seattle, WA 98195
lubetkin@uw.edu
grace maze
RSMAS, University of Miami
3894 Grand Ave
Miami, FL 33133
gmaze@rsmas.miami.edu
Susanne menden-deuer
University of Rhode Island
Graduate School of Oceanography
South Ferry Road
Narragansett, RI 02882
smenden@mail.uri.edu
nina neill
ninaeneill@gmail.com
Kevin meyer
University of Maryland Center for
Environmental Science
2020 Horns Point Road
Cambridge, MD 21613
kmeyer@umces.edu
harry nelson
Fluid Imaging Technologies
65 Forest Falls Drive
Yarmouth, Maine 04096
harry@fluidimaging.com
elizabeth miller
Florida Department of Environmental
Protection
2600 Blairstone Road, MS6515
Tallahassee, FL 32399
elizabeth.miller@dep.state.fl.us
PARTICIPANT LIST
david millie
Palm Island Enviro-Informatics/
Michigan Technological University
4645 Stone Ridge Trail
Sarasota, FL 34232
dfmillie@mtu.edu
Sarah milton
777 Glades Road, SC 288
smilton@fau.edu
Stephanie moore
NOAA Fisheries
Northwest Fisheries Science Center
2725 Montlake Boulevard East
Seattle, WA 98112
stephanie.moore@noaa.gov
Page 218
ari nissanka
Sarasota, FL 34236
ari@mote.org
Susan niven
nivens@uncw.edu
Sheila o’dea
Commission
100 8th Ave SE
sheila.odea@myfwc.com
Sean o’mara
Texas A&M University - Corpus Christi
6300 Ocean Drive, Unit 5866
somara@islander.tamucc.edu
Kevin o’Shea
11200 SW 8th Street
Miami, FL 33174
osheak@fiu.edu
timothy otten
Oregon State University
220 Nash Hall
Corvallis, OR 97331
ottent@onid.orst.edu
Kevin owen
Port Dolphin Energy, LLC
Höegh LNG AS
400 North Tampa Street, Suite 1015
Tampa, FL 33602
kowen1972@yahoo.com
hans paerl
University of North Carolina at Chapel Hill
Institute of Marine Sciences
3431 Arendell Street
Morehead City, NC 28557
hpaerl@email.unc.edu
valeriy palubok
Sarasota, FL 34236
palubok@mote.org
Kevin petrus
Florida Department of Environmental
Protection
2600 Bairstone Road
kevin.petrus@dep.state.fl.us
Jennifer phillips
University of Maine
5735 Hitchner Hall, Rm. 297
Orono, ME 04469
jennifer.phillips@umit.maine.edu
richard pierce
Sarasota, FL 34236
rich@mote.org
Kathleen pitz
266 Woods Hole Road
kpitz@whoi.edu
allen place
UMCES-IMET
701 East Pratt Street
Baltimore, MD 21202
shorn@umces.edu
Kaytee pokrzywinski
700 Pilottown Rd
Lewes, DE 19958
kaytee@udel.edu
Katherine perri
State University of New York - College of
Environmental Science and Forestry
1 Forestry Drive, 121 Jahn Lab
Syracuse, NY 12310
katherineaperri@gmail.com
helena pound
University of Tennessee Knoxville
M409 Walters Life Sciences
Knoxville, TN 37996
hpound@utk.edu
Page 219
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PARTICIPANT LIST
michael parsons
mparsons@fgcu.edu
pete Quasius
Collier County Audubon
4523 E Riverside Drive
pquasius@msn.com
regina radan
1156 High Street
rradan@ucsc.edu
John ramsdell
NOAA-National Ocean Service
john.ramsdell@noaa.gov
PARTICIPANT LIST
Carlton rauschenberg
Bigelow Laboratory
P.O. Box 380
East Boothbay, ME 04544
carlton@bigelow.org
andrew reich
Florida Department of Health
4052 Bald Cypress Way
Bin A12
andy_reich@doh.state.fl.us
tammi richardson
University of South Carolina
Dept. of Biological Sciences and
Marine Science Program
Columbia, SC 29208
richardson@biol.sc.edu
mindy richlen
266 Woods Hole Road, MS32
mrichlen@whoi.edu
alison robertson
University of South Alabama
Department of Marine Sciences
Mobile, AL 36606
arobertson@disl.org
anne rolton
10501 FGCU Blvd, South
annerolton@hotmail.co.uk
Katrin rudge
Riverview High School
1 Ram Way
Sarasota, FL 34231
katrin.rudge@sarasotacountyschools.net
Kelly rein
11200 SW 8th Street
Miami, FL 33199
reink@fiu.edu
John ryan
Monterey Bay Aquarium Research Institute
7700 Sandholdt Road
ryjo@mbari.org
Bill richardson
Commission
100 8th Ave SE
bill.richardson@myfwc.com
darcie ryan
deryan@geos.tamu.edu
Page 220
marci Savage
State University of New York
Environmental Science and Forestry School
155 Hood Ave
Syracuse, NY 13208
mlsavage@syr.edu
dianne Shipley
Florida Department of Health in Sarasota
County
2200 Ringling Boulevard
Sarasota, FL 34237
dianne_shipley@doh.state.fl.us
Justine Schmidt
State University of New York
College of Environmental Science and
Forestry
341 Jahn Lab, 1 Forestry Drive
Syracuse, NY 13210
jrschmid@syr.edu
Sandra Shumway
University of Connecticut
Department of Marine Sciences
1080 Shennecossett Road
Groton, CT 06340
patricia.evans@uconn.edu
Julius Schneider
East Georgia State College
131 College Circle
Swainsboro, GA 30401
jschneid@ega.edu
Chris Simoniello
GCOOS
140 7th Avenue South
simo@marine.usf.edu
emily Smith
Louisiana State University
1239 Energy, Coast and Environment
Building
Baton Rouge, LA 70803
esmi122@lsu.edu
erica Seubert
University of Southern California
3616 Trousdale Parkway
Los Angeles, CA 90089
seubert@usc.edu
g. Jason Smith
Moss Landing Marine Laboratories
8272 Moss Landing Road
jsmith@mlml.calstate.edu
Sugandha Shankar
University of Maine
School of Marine Sciences
5706 Aubert Hall
Orono, ME 04469
sugandha.shankar@maine.edu
Jayme Smith
3616 Trousdale Parkway, AHF 301
jaymesmi@usc.edu
Justin Shapiro
Sarasota, FL 34236
shapiro@mote.org
Juliette Smith
266 Woods Hole Road
jlsmith@whoi.edu
Page 221
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October 2013
PARTICIPANT LIST
Kevin Sellner
Chesapeake Research Consortium
645 Contees Wharf Road
Edgewater, MD 21037
sellnerk@si.edu
Stephanie Smith
Beagle Bioproducts, Inc.
959 Schrock Road
Columbus, OH 43229
stephanie.smith@beaglebioproducts.com
dalton Steele
3000 N.E. 151st Street
dstee002@fiu.edu
morgan Steffen
University of Tennessee
M409 WLS
Knoxville, TN 37996
morgan.m.steffen@gmail.com
PARTICIPANT LIST
Karen Steidinger
Commission
100 8th Ave SE
karen.steidinger@myfwc.com
William Stringfellow
University of the Pacific-Ecological
Engineering Research Program
3601 Pacific Avenue
Chambers Tech Center Rm 224
Stockton, CA 95211
wstringfellow@lbl.gov
Jamie Studts
Department of Behavioral Science
University of Kentucky
127 Medical Behavioral Science Building
Lexington, KY 40536
jlstu2@email.uky.edu
rick Stumpf
NOAA NCOS
1305 East-West Highway
Silver Spring, MS 20902
richard.stumpf@noaa.gov
Page 222
marc Suddleson
DOC/NOAA/NOS/CSCOR
1305 East-West Highway, Rm 8254
Silver Spring, md 20910
marc.suddleson@noaa.gov
pengfei Sun
11200 SW 8th Street
Miami, Fl 33199
psun001@fiu.edu
Karen Swajian
5100 Paint Branch Parkway
College Park, MD 20740
karen.swajian@fda.hhs.gov
avery tatters
3616 Trousdale Parkway
tatters@usc.edu
pat tester
101 Pivers Island Road
Beaufort, NC 28516
patricia.tester@noaa.gov
Kristen thyng
3146 TAMU
kthyng@tamu.edu
Charles tilney
700 Pilottown Rd.
Lewes, DE 19958
ctilney@gmail.com
mengmeng tong
Zhejiang University/Woods Hole
Oceanographic Institution
266 Woods Hole Road, MS32
mtong@whoi.edu
Kevin tyre
Fort Myers, NY 33965
kntyre@eagle.fgcu.edu
Steven ullmann
The University of Miami Health Sector
Management and Policy
P.O. Box 248442
sullmann@bus.miami.edu
Cristina urizar
NOAA/NOS/CO-OPS
1305 East West Highway, Station 7124
cristina.urizar@noaa.gov
Kathryn van alstyne
Western Washington University
1900 Shannon Point Road
Anacortes, WA 98221
kathy.vanalstyne@wwu.edu
mark.vanasten@diagnostictechnology.com.au
frances van dolah
NOAA Center for Coastal Environmental
Health and Biomolecular Research
fran.vandolah@noaa.gov
mark Warner
700 Pilottown Road
Lewes, DE 19958
mwarner@udel.edu
rhonda Watkins
Collier County Governemnt
3327 Tamiami Trail East
Naples, FL 34112
rhondawatkins@colliergov.net
Cathy Wazniak
Maryland Dept of Natural Resources
580 Taylor Ave, D2
Annapolis, MD 21401
cwazniak@dnr.state.md.us
Samantha Weber
State University of New York College of
Environmental Science and Forestry
816 Maryland Ave
Syracuse, NY 13210
saweber@syr.edu
Ben Whitenack
Mote Marine Lab
Sarasota, FL 34236
whiteben@sas.upenn.edu
Page 223
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October 2013
Cathy Walsh
Sarasota, FL 34236
cjwalsh@mote.org
PARTICIPANT LIST
mark van asten
Diagnostic Technology Pty Ltd.
Suite 45, 7 Narabang Way
Belrose, NSW, 2085
Australia
Jennifer vreeland
Mote Marine Laboratories
Sarasota, FL 34236
jvreeland@mote.org
Steven Wilhelm
Department of Microbiology
The University of Tennessee
Knoxville, TN 37996
wilhelm@utk.edu
lori zaworski
1600 Ken Thompson Pkwy.
Sarasota, FL 34236
lori@mote.org
april Woods
Moss Landing Marine Labs
8272 Moss Landing Road
awoods@mlml.calstate.edu
Bingxue zheng
11200 SW 8th Street
Miami, FL 33199
bzhen003@fiu.edu
Jennifer yordy
1600 Ken Thompson Pkwy
Sarasota, FL 34236
jyordy@mote.org
paul zimba
Center for Coastal Studies
6300 Ocean Drive Unit 5866
paul.zimba@tamucc.edu
lawrence younan
Turner Designs, Inc.
845 West Maude Avenue
Sunnyvale, CA 94085
lyounan@turnerdesigns.com
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rich zamor
University of Oklahoma
1033 W. Boyd St.
Norman, OK 73069
rmzamor@ou.edu
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