An An in situin situ Sensor of Sensor of Phytoplankton Community Phytoplankton Community Structure Based on Light Structure Based on Light

AbsorptionAbsorption

Gary Kirkpatrick, David Millie, Gary Kirkpatrick, David Millie, Steven Lohrenz, Mark Moline, Steven Lohrenz, Mark Moline,

Ian Robbins and Oscar SchofieldIan Robbins and Oscar Schofield

AcknowledgementsAcknowledgements

National Science FoundationNational Science Foundation– Biological Sciences DirectorateBiological Sciences Directorate– Ocean Sciences DivisionOcean Sciences Division

National Oceanic and Atmospheric AdministrationNational Oceanic and Atmospheric Administration– ECOHABECOHAB– MERHABMERHAB– Sea GrantSea Grant

Florida Fish and Wildlife Conservation Florida Fish and Wildlife Conservation CommissionCommission– Florida Fish and Wildlife Research InstituteFlorida Fish and Wildlife Research Institute

Office of Naval ResearchOffice of Naval Research

Finding, Tracking and Finding, Tracking and Mapping Harmful AlgaeMapping Harmful Algae

Our Approach to HAB Detection, Our Approach to HAB Detection, Tracking and MappingTracking and Mapping

Based on Based on in situin situ particulate absorption particulate absorption spectraspectraOriginally, analysis by similarity of 4Originally, analysis by similarity of 4thth derivative spectraderivative spectraMost recently, multiple species regression Most recently, multiple species regression analysis (after Stæhr and Cullen) analysis (after Stæhr and Cullen)

Advantages Over (traditional) Advantages Over (traditional) Microscopic EnumerationMicroscopic Enumeration

Readily automated – minimal human Readily automated – minimal human involvementinvolvement

Good spatial and temporal coverageGood spatial and temporal coverage

Less weather dependentLess weather dependent

EconomicalEconomical

This PresentationThis Presentation

Single-species SimilaritySingle-species Similarity– laboratory to autonomous field applicationlaboratory to autonomous field application

Multiple-species Community CompositionMultiple-species Community Composition– laboratory to field trialslaboratory to field trials

FO

Spectrometer

Light Source

LWCCFO

ReferenceWater

Cross-flow Filter

Valve

Cleaning Solutions

Pump

Valve Valve

Optical Phytoplankton DiscriminatorOptical Phytoplankton Discriminator(OPD)(OPD)

Ch

l cC

hl b

ph

cob

ilin

Laboratory Class ComparisonsLaboratory Class Comparisons

Shipboard Underway HAB DetectionShipboard Underway HAB Detection

OPD Map of HABOPD Map of HAB

0.25

0.315

0.38

0.445

0.51

0.575

0.64

0.705

0.77

0.835

0.9

-83.4 -83.2 -83 -82.8 -82.6 -82.426.6

26.8

27

27.2

27.4

27.6

Karenia brevis Cell Count

(cells l-1

)

0 to 20,000

20,000 to 200,000

200,000 to 368,000

368,000 to 670,000

670,000 to 1,421,000

Sarasota

TampaBay

Karenia brevisSimilarity Index

OPD-equipped VehiclesOPD-equipped Vehicles

REMUS – Propeller Driven

Glider – Buoyancy Driven

BreveBuster Payload

BSOP Vertical Profiler

Launch

Before Dive

Gliding

OPD-equipped Glider Surveying HABOPD-equipped Glider Surveying HAB

Mission Track& Satellite Image

Resulting Red Tide Distribution

Glider Mission - Sep 28, 2004 – Oct 7, 2004Glider Mission - Sep 28, 2004 – Oct 7, 2004

Karenia brevisKarenia brevis Similarity Contours Similarity Contours

-83.05 -83 -82.95 -82.9 -82.85 -82.8 -82.75 -82.7-30

-25

-20

-15

-10

-5

0

-0.35

-0.25

-0.15

-0.05

0.05

0.15

0.25

0.35

Fitting Multiple ClassesFitting Multiple Classes

Least squares, multiple regression analyses (after Least squares, multiple regression analyses (after Stæhr and Cullen, 2003).Stæhr and Cullen, 2003).

Optimization/reduction algorithm to minimize Optimization/reduction algorithm to minimize computational load.computational load.

Laboratory Species MixesLaboratory Species MixesOPD OPD vsvs CHEMTAX CHEMTAX

Gyrodinium instriatum

y = 1.1951x - 11.348

R2 = 0.5241

-20

0

20

40

60

80

100

120

140

0.0 20.0 40.0 60.0 80.0

Chlorophyll a calculated by CHEMTAX

Ch

loro

ph

yll

a ca

lcu

late

d b

y B

reve

Bu

ster

GI

Linear (GI)

Tetraselmis impellucida

y = 0.8859x + 0.2528

R2 = 0.7526

050

100150200250300350400450500

0.0 100.0 200.0 300.0 400.0 500.0

Chlorophyll a calculated by CHEMTAX

Ch

loro

ph

yll

a ca

lcu

late

d b

y B

reve

Bu

ster

TI

Linear (TI)

Karenia brevis

y = 0.8809x + 5.3908

R2 = 0.8816

0102030405060708090

100

0.0 50.0 100.0 150.0

Chlorophyll a calculated by CHEMTAX

Ch

loro

ph

yll

a ca

lcu

late

d b

y B

reve

Bu

ster

KB

Linear (KB)

Dactyliosolen fragilissimus

y = 0.2787x + 4.9817

R2 = 0.0092

0123456789

10

-1.0 0.0 1.0 2.0 3.0 4.0

Chlorophyll a calculated by CHEMTAX

Ch

loro

ph

yll

a ca

lcu

late

d b

y B

reve

Bu

ster

DF

Linear (DF)

Multiple Class LibraryMultiple Class Library

Species Source Division Class IDTrichodesmium sp. Rutgers Univ, IMCS Cyanophyta Cyanophyceae Cyn_cyan_1Dunaliella tertiolecta Rutgers Univ, IMCS Chlorophyta Chlorophyceae Chl_chlo_1Emilinia huxleyi Rutgers Univ, IMCS Haptophyta Prymnesiophyceae Hap_prym_1Skeletonema costatum Rutgers Univ, IMCS Heterokonta Bacillariophyceae Het_baci_1Thalassira weissflogii Rutgers Univ, IMCS Heterokonta Bacillariophyceae Het_baci_2Phacodactylum tricronutumRutgers Univ, IMCS Heterokonta Bacillariophyceae Het_baci_3Prorocentrum minimum Rutgers Univ, IMCS Dinophyta Dinophyceae Din_dino_1Heterosigma akashiwo Rutgers Univ, IMCS Heterokonta Raphidophyceae Het_Raph_1Tetraselmis sp. Rutgers Univ, IMCS Chlorophyta Prasinophyceae Chl_Pras_1Isochrysis galbana Rutgers Univ, IMCS Haptophyta Prymnesiophyceae Hap_prym_2Karenia brevis Mote Marine Lab Dinophyta Dinophyceae Kb_041111

Library Standard SpectraLibrary Standard Spectra

Absorbance Fourth Derivative

Natural Community ClassesNatural Community ClassesOPD Chl OPD Chl aa vsvs CHEMTAX Chl CHEMTAX Chl aa

Prymnesiophyceae Component

y = -0.2215x + 1.7612R2 = 0.0283

0

0.5

1

1.5

2

2.5

3

0 1 2 3

CHEMTAX Chl a (ug/l)

OP

D C

hl a

(u

g/l)

Cyanophyceae Component

y = 0.3083x - 0.0391R2 = 0.0374

0

0.5

1

1.5

2

0 0.5 1 1.5 2

CHEMTAX Chl a (ug/l)

OP

D C

hl a

(u

g/l)

Bacillariophyceae Component

y = 0.4301x + 0.6429R2 = 0.4447

0

0.5

1

1.5

2

2.5

3

0 0.5 1 1.5 2 2.5 3

CHEMTAX Chl a (ug/l)

OP

D C

hl a

(u

g/l)

Chlorophyceae Component

y = -0.2451x + 0.8897R2 = 0.4847

0

1

2

3

4

5

0 1 2 3 4 5

CHEMTAX Chl a (ug/l)

OP

D C

hl a

(u

g/l)

Dinophyceaea Component

y = 0.4673x - 0.1295R2 = 0.2792

0

2

4

6

8

10

0 2 4 6 8 10

CHEMTAX Chl a (ug/l)

OP

D C

hl a

(ug

/l)

Total Chlorophyll a

y = 0.9837x - 0.406R2 = 0.5101

5

6

7

8

9

10

11

12

5 6 7 8 9 10 11 12

CHEMTAX Chl a (ug/l)O

PD

Ch

l a (

ug

/l)

DiscussionDiscussion

Initial results encouraging.Initial results encouraging.

Library of ‘standard’ species incomplete!Library of ‘standard’ species incomplete!

Libraries need to be regional.Libraries need to be regional.

Culture ID and condition are critical!!Culture ID and condition are critical!!– Garbage in, garbage out!Garbage in, garbage out!