CALIFORNIA MARINE

BIOTOXINS:

Overview of Statewide

Monitoring Program

Marine Biotoxin Monitoring Program

California Department of Public HealthEnvironmental Management Branch

Gregg Langlois

Marine Biotoxin BackgroundMarine Biotoxin Background

Naturally occurring: All Climatic Regimesnerve toxins: nuisance to deadlyproduced by a few phytoplankton species

Dinoflagellates: PSP, NSP, DSPDiatoms: ASP

Bioconcentrated: food chain transportNo antidotes (including garlic butter, EtOH)

Marine Biotoxin BackgroundMarine Biotoxin Background

Regions with History of OutbreaksRegions with History of OutbreaksRegions with only Recent OutbreaksRegions with only Recent Outbreaks

GuatamalaGuatamala: 1987, 26 deaths (PSP; P. : 1987, 26 deaths (PSP; P. bahamensebahamense))PhillipinesPhillipines: 1983, 21 deaths (PSP): 1983, 21 deaths (PSP)Canada: 1987, 4 deaths (ASP)Canada: 1987, 4 deaths (ASP)New Zealand: 1993, 180 cases (NSP, PSP)New Zealand: 1993, 180 cases (NSP, PSP)

Global Increase in Frequency and DistributionGlobal Increase in Frequency and Distribution

Marine Biotoxin BackgroundMarine Biotoxin Background

Paralytic Shellfish Poisoning (PSP) ToxinsDinoflagellate: Alexandrium catenellaProgram (field, lab, management) built around this one toxin

Amnesic Shellfish Poisoning (ASP) = Domoic Acid

Diatom: Pseudo-nitzschia spp.Overnight challenge to existing program: field, lab, management

Alexandrium

Pseudo-nitzschia

PSP Health Effects

Sodium Channel BlockerAlert Level: ≥ 80 ug/100 g tissueMild Symptoms: 200 - 400 ug

(numbness of extremities, headache)Moderate/Severe: 500 - 2000 ug

(loss of coordination, difficulty breathing)Severe Symptoms: > 2000 ug

(paralysis, respiratory failure, death)

1927 – 1936:PSP toxin assay begins using newly developed mouse bioassay.

PSP Assay: Mouse Bioassay

HN

R4

R1

R2R3

O

ON N

NHNH N+

2

NH +2

H

OHOH

Saxitoxin

2007:Only approved method (FDA)Low sensitivity (40 ug/100 g)Labor intensiveRequires large quantities of materialsLimited to central lab concept

D.A. Health Effects

Neuroexcitatory Amino Acid (glutamate receptors)Alert Level: 20 ug/g (ppm)Mild Symptoms: 27 - 75 ppm

(GI: cramps, diarrhea, vomiting)

Moderate : 40 - 700 ug(headache, disorientation)

Severe Symptoms: > 450 ug(loss of short-term memory, seizure, coma, death)

DA Analysis: HPLCHigh capital expenseSimple methodology

Methanol extractionSmall sample sizeTransferable to other labs

Automated: rapid throughputLow detection limit (~ 1 ppm)

M. A. Quilliam. Chemical methods for domoic acid, the amnesic shellfish poisoning (ASP) toxin. In: G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae, Monographs on Oceanographic Methodology, Vol. 11, Chapter 9. Intergovernmental Oceanographic Commission (UNESCO), Paris, 247-266 (2003).

R

E

Z

Alternative Analytical MethodsDomoic Acid

Immunoassays (Biosense, NOAA, Jellett)Jellett test is qualitative (presence/absence)

PSP ToxinsLawrence HPLC MethodReceptor Binding AssayImmunoassay (Jellett)

Shuck musselsShuck musselsMolluscoMollusco--maticmatic

ExtractExtract(acidified IPA)(acidified IPA)

FilterFilterJellett Rapid Test: PSPJellett Rapid Test: PSP

Wait 30 minutesWait 30 minutesAdd extract to Add extract to

bufferbufferAdd buffer to stripAdd buffer to strip

1. Plankton Sampling 2. Plankton observations

3. Decision: run tests on plankton, shellfish?4. Decision: ship samples to lab?

Monitoring DataMonitoring DataShellfish

Traditional ApproachTesting Food for ToxinTide, Weather DependentLow Frequency, High Cost

PhytoplanktonAdvanced warningRelation to toxicityNot limited by tide, weatherImmunoassay (Jellett)

InterannualInterannual VariabilityVariabilityPSP Toxicity in California: 1981 - 2006

0

500

1000

1500

2000

2500

3000

3500

4000

4500

5000

1/1/

1981

1/1/

1982

1/1/

1983

1/1/

1984

1/1/

1985

1/1/

1986

1/1/

1987

1/1/

1988

1/1/

1989

1/1/

1990

1/1/

1991

1/1/

1992

1/1/

1993

1/1/

1994

1/1/

1995

1/1/

1996

1/1/

1997

1/1/

1998

1/1/

1999

1/1/

2000

1/1/

2001

1/1/

2002

1/1/

2003

1/1/

2004

1/1/

2005

1/1/

2006

Date

PSP

Toxi

city

(ug/

100

g)

10,000 ug14,000 ug

Domoic Acid Concentrations in California: 1991 - 2006

0

50

100

150

200

250

300

350

1/1/

1991

1/1/

1992

1/1/

1993

1/1/

1994

1/1/

1995

1/1/

1996

1/1/

1997

1/1/

1998

1/1/

1999

1/1/

2000

1/1/

2001

1/1/

2002

1/1/

2003

1/1/

2004

1/1/

2005

1/1/

2006

Date

Dom

oic

Aci

d (p

pm)

Biological Cues:Phytoplankton

Photo courtesy of Kai Schumann

“RED TIDES”“RED TIDES”Vast majority are nontoxic

Anecdotal health effectsWildlife impactsEconomic impacts“Harmful Algal Blooms (HABs)”

Alexandrium rarely “blooms” Photo courtesy of Tara Crow, Santa Monica Pier Aquarium

Phytoplankton Monitoring

Ceratium

Easy to do in the field:Plankton net, field microscopeToxic species easily to IDRapid throughputLow cost: Volunteer-basedNot weather/tide dependent

Alexandrium

Chaetoceros sp.

Alexandrium: Seasonal Relative Abundance Variability(1993 - 2006)

0

1

2

3

4

5

6

7

8

9

10

1 29 55 83 110 136 163 187 209 232 255 281 306 334 362

Days (Julian)

Rel

ativ

e A

bund

ance

Inde

x

Phyto Data:-Alexandrium:

-Not usually present

-Low #s = Toxicity

-Presence triggers response

Phyto Data:-Pseudo-nitzschia:

-Often present

-Track increases

-Monospecific blooms

-Increasing #s triggers response

Pseudo-nitzschia: Seasonal Variability in Relative Abuncance Index(1993 - 2005)

0

10

20

30

40

50

60

70

80

90

100

1 18 33 46 61 73 86 96 105

114

122

131

140

149

158

166

176

185

195

204

212

221

229

239

248

258

268

281

293

303

315

326

340

354

Days (Julian)

Rel

ativ

e A

bund

ance

Inde

x

Environmental Cues:Remote Sensing Data: Buoys

Environmental Cues15 Day Upwelling Indices: 1989 - 2005

-400

-300

-200

-100

0

100

200

300

400

01/0

1/89

01/0

1/90

01/0

1/91

01/0

1/92

01/0

1/93

01/0

1/94

01/0

1/95

01/0

1/96

01/0

1/97

01/0

1/98

01/0

1/99

01/0

1/00

01/0

1/01

01/0

1/02

01/0

1/03

01/0

1/04

01/0

1/05

Date

Upw

ellin

g In

dex

39N125W

Upwelling Index

Sea Surface Temperatures: SF Buoy

8.0

10.0

12.0

14.0

16.0

18.0

20.0

1/1/

1989

1/1/

1990

1/1/

1991

1/1/

1992

1/1/

1993

1/1/

1994

1/1/

1995

1/1/

1996

1/1/

1997

1/1/

1998

1/1/

1999

1/1/

2000

1/1/

2001

1/1/

2002

1/1/

2003

1/1/

2004

Year

Tem

pera

ture

(°C

) Sea Surface Temperature

(SST)

Environmental Cues:Environmental Cues:Remote Sensing Data: Satellites

Sea Surface Temperatures via AVHRR: Advanced Very High ResolutioSea Surface Temperatures via AVHRR: Advanced Very High Resolution Radiometryn Radiometry

8.89.6

10.411.212.012.813.614.415.216.016.817.618.419.220.0

215 220 225 230 235 240 245 250DAYS (Julian)

100

200

300

PSP

TOXI

NS

(ug)

August 15, 1994

August 25, 1994

8.89.6

10.411.212.012.813.614.415.216.016.817.618.419.220.0

215 220 225 230 235 240 245 250DAYS (Julian)

100

200

300

PSP

TOXI

NS

(ug)

September 3, 1994

8.89.6

10.411.212.012.813.614.415.216.016.817.618.419.220.0

215 220 225 230 235 240 245 250DAYS (Julian)

100

200

300

PSP

TOXI

NS

(ug)

Environmental Cues:Environmental Cues:Marine Mammal Strandings

Symptoms typical of Domoic Acid poisoning (seizures)

Integrated Monitoring: 2009Environmental Data:

Offshore Buoy SSTImagery: AVHRR and SeaWifsMarine mammal strandingsBloom transport tracking models

Phytoplankton DataField sampling, field/lab observations: toxic speciesField test kits: cell toxicityGene probes: toxic species

Shellfish Toxicity DataImproved lab methods: automated, rapid, nonproprietary

Field screening for toxins (cull negatives; harvest info)

Photo courtesy of Todd Warshaw

Acknowledgements:Del Norte County Health DepartmentU.C. Sea Grant ExtensionCoast Seafoods CompanyHumboldt Co. Environmental Health Dept.CDHS VolunteersMendocino Co. Environmental Health Dept.CDHS VolunteerCalifornia Department of Fish and GameCDHS VolunteersCordell Banks National Marine SanctuaryBodega Marine LabJohnson Oyster CompanyDrakes Bay Oyster CompanyHog Island Oyster CompanyMarin Oyster CompanyCove Mussel CompanyCDHS/UCSC Cal-PReEMPTThe Marine Mammal Center (TMMC)San Francisco County Health DepartmentGulf of the Farallones Nat. Marine SantuaryCDHS VolunteersSan Mateo Co. Environmental Health Dept.TMMC/CDHS VolunteersSanta Cruz Co. Environmental Health Dept.U.C. Santa Cruz (UCSC)CDHS/UCSC Cal-PReEMPTTMMC/CDHS VolunteersPacific Cetacean GroupCDHS VolunteersMonterey Abalone CompanyTMMC/CDHS VolunteersMarine Pollution Studies LaboratoryNOAA Coastal Discovery CenterWilliams Shellfish CompanyTomales Bay Oyster Company (Morro Bay)

Morro Bay National Estuary ProgramMorro Bay Natural History MuseumCalifornia Department of Fish and GameTenera EnvironmentalTMMC/CDHS VolunteersCDHS/UCSC Cal-PReEMPTVandenberg AFBCalif. Department of Parks and RecreationU.C. Santa BarbaraSanta Barbara City CollegeSanta Barbara Mariculture CompanyEcomar, Inc.Santa Barbara Channel KeeperNational Park ServiceChannel Island National Marine SanctuarySB CDHS VolunteersVentura Co. Environmental Health Dept.Pt. Mugu NASVentura CDHS VolunteersLos Angeles Co. Health Dept.Los Angeles County Sanitation DistrictCity of L.A. Environmental Monitoring Div.Regional Water Quality Control BoardCatalina Island Marine InstituteCatalina Tall Ships ExpeditionsUniversity of Southern CaliforniaAquarium of the Pacific, Long BeachL.A. CDHS VolunteersOrange County Sanitation DistrictOcean InstituteEcomar, Inc.Orange CDHS VolunteersSan Diego CDHS VolunteersCarlsbad Aquafarms Inc.Scripps Institute of OceanographyAvian Research AssociationU.S. Navy