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Aureococcus anophagefferensDelaware Inland Bays 1998 - 2005

PartnersAcademy of Natural Sciences – Ann Marie Hartsig, Richard Lacouture

Maryland DNR – Walt Butler

University of Delaware, College of Marine Studies – Kathy Coyne, Linda Popels, Dave Hutchins

University of Maryland, Baltimore – Holly Bowers, Dave Oldach

[email protected]

DE STAC 3/31/06NOT FOR REPRODUCTIOWITHOUT CONSENT OFAUTHOR

OutlineAnalysis Method to ID and enumerate Aureococcus

Aureococcus cell densities across sampling period and stations

Aureococcus maximum cell density per station

Aureococcus frequency of occurrence per station

Aureococcus cell density per date per station versus water temperature and salinity

Recap

NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR

Aureococcus anophagefferens Analysis MethodsDelaware Inland Bays

U MD Baltimore (Bowers)2004

Qualitative PCR (Preserved Sample)

Variable with physiological state of target cell

MD DNR (Butler)2002Light Microscopy (Live Sample)using Transmitted Light

UD CMS (Coyne/Popels)2001Quantitative PCR (Live Sample)

Variable with physiological state of target cell

ACNS (Hartsig)2002

Suffolk County, NY, Dept. of Health Services1998

Polyclonal Antibody & Epifluorescence microscopy

May underestimate Aureococcus cell density by factor of 2 to 3 (D. Caron, communication)

Laboratory (Researcher)YearAnalytical Method


0.0E+00

2.0E+07

4.0E+07

6.0E+07

8.0E+07

1.0E+08

1.2E+08

1.4E+08

6/24/9

86/2

4/98

5/22/0

15/2

2/01

5/22/0

16/1

1/01

6/11/0

16/1

1/01

6/26/0

16/2

6/01

6/26/0

17/1

1/01

7/11/0

17/1

1/01

7/25/0

17/2

5/01

7/25/0

18/6

/018/6

/018/6

/018/2

9/01

8/29/0

18/2

9/01

6/10/0

26/2

4/02

7/8/02

7/22/0

28/6

/028/2

1/02

9/10/0

210

/1/02

7/1/04

7/13/0

47/2

7/04

8/10/0

48/2

5/04

9/8/04

Date

DE Ditch LA-8

-- - - - - - - - - - - - - - - - - - - - - - - - - - Present - - - - - - - - - - - - - - - - - -

Pre

sent

Aureococcus (cells/L)Delaware Little Assawoman Bay: 1998, 2001, 2002, 2004

(Site location variable by year)

Collection Dates NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR

PCR qual. positiveLA-8The Ditch, LABJuly2004

polyclonal139,976,000LA-8The Ditch, LABlive screening82,000,000LA-8The Ditch, LAB

June2002

PCR quant. 213RLC-3Love Creek, RBSept.

PCR quant. 40,248LA-1Little Assawoman Bay - open bayPCR quant. 3,429IP-2

Pepper Creek, Indian River/Bay (IRB)

PCR quant. 1,470IR-4Indian RiverPCR quant. 15,758IR-1Indian River

August

PCR quant. 189,034LA-6Dirickson Creek, LAB

PCR quant. 29,492LA-5Miller Creek, LABJune

PCR quant. 821,548TC-1Torquay Canal, RBPCR quant. 58,807RLC-2Love Creek, RBPCR quant. 3,846AC-2Arnell Creek, Rehoboth Bay (RB)

May

2001

polyclonal743,000LA-4Dirickson Creek, LAB

polyclonal108,000LA-2Little Assawoman Bay (LAB)June1998

Analytical Method#cells/LStationWaterbodyMonthYear

Aureococcus anophagefferens (Maximum Cell Density)


0.E+00

2.E+07

4.E+07

6.E+07

8.E+07

1.E+08

1.E+08

1.E+08

2.E+08

20 22 24 26 28 30 32

6/10/02, LA-8

6/24/02, LA-8

7/8/02, LA-8

Aureococcus (cells/L) vs. Water Temperature οCDelaware Little Assawoman Bay – multiple years


0.E+00

2.E+07

4.E+07

6.E+07

8.E+07

1.E+08

1.E+08

1.E+08

2.E+08

0 4 8 12 16 20 24 28 32

6/10/02, LA-8

6/24/02, LA-8

7/8/02, LA-8

Aureococcus (cells/L) vs Salinity (ppt)Delaware Little Assawoman Bay – multiple years


RECAP: Delaware Inland Bays

Only 1 in 113 samples tested for Aureococcus showed densities of bloom magnitude (approximately 100,000,000 cells/L )

NO sample exhibited a cell density equivalent to a Maryland Category 3 Bloom (> 200,000,000 cells/L, polyclonal technique)

Cell density varies by analysis method

Cell density was compared to concentrations of selected environmental parameters (Urea, TN, DNH3, DOC, DON/DIN, TDN, TP) ---No significant relationship - sample size small and varied: 58 – 68

Highest concentration was reported from “The Ditch”, which joins Delaware Little Assawoman Bay to Maryland Assawoman Bay


Aureococcus anophagefferens

A glimpse !

Delaware Little Assawoman Bay

and

Maryland Assawoman Bay

Results: 2001, 2002, and 2004

Edythe Humphries, Delaware DNREC

DELAWARE STAC 3/31/06


Outline

Little Assawoman Bay & Assawoman Bay Vital Facts

Sampling Sites in DE and MD

Analysis Methods: DE and MD

Cell Densities and Surface Water Temperature: DE and MD

Cell Densities and Surface Salinity: DE and MD

Cell Densities: DE and MD2001200220042001 – 2004 at “The Ditch”

General Observations

Monitoring Activities in 2006: DE and MD NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR

Little Assawoman Bay (LAB) Vital Facts

Water Surface Area: ~ 4 sq. miles …… 14% of watershed

Groundwater Contribution: yes, to bay shore and as stream baseflow

Watershed Size (2002 record) : 37 sq. miles (35% agriculture, 20% wetlands, 19% urban, 10% forest, 2% barren/range, 14% water)

Residence Time:currently unknownTidal Influence: From North (Assawoman Canal) – max. 5 cm/sec at Canal

From South (Ocean City Inlet) – ~ 50cm/sec at The Ditch(wind driven: max. 10 cm/sec)

Eelgrass: none reportedWidgeon grass: isolated stands

Shellfish:ribbed mussel - presenthard clams – historically, currently limited natural set, currently an active gardening programoysters – currently an active gardening program (good oyster growth)scallops – none reported


Assawoman Bay (AB) Vital Facts

Water Surface Area: 9.2 sq. miles . . . . . . . 85% of watershed

Groundwater Contribution: yes, 35% of Nitrogen source

Watershed Size (2002 record) : 10.7 sq. miles (23.7% agriculture, 21.4% wetlands, 28.3% urban, 25% forest, 1.6% barren/beaches, 85% water)

Residence Time:20.9 days

Tidal Influence: From South (Ocean City Inlet)

SAV (Eelgrass, Widgeon grass, etc.) : 8% of bay bottom most prevalent in south end of Bay

Shellfish:hard clams – average of 0.18 live clams/m2 (2003) – low compared to other

Maryland Coastal Baysscallops – reported in 2002


ComparisonDelaware Little Assawoman Bay (LAB) versus Maryland Assawoman Bay (AB)

RPD = Relative Percent Difference

• 110 RPD – Watershed Size – LAB larger %

• 38 RPD – Agriculture/Watershed – LAB larger %

• 143 RPD – Surface Water/Watershed – AB larger %

• 85 RPD – Forest/Watershed – AB larger %

• 39 RPD – Urban/Watershed – AB larger %


Dave CaronMonoclonal Antibody & ELISA Low cross-reactivity with other protists & bacteria

U MD Baltimore (Bowers)

Qualitative PCR (Preserved Sample -DNA) Variable with physiological state of target cell

2004

MD DNR (Butler)MD DNR (Butler)Light Microscopy (Live Sample)

ACNS (Hartsig)ACNS (Hartsig)Polyclonal Antibody & Fluorescence microscopy

2002

UD CMS (Coyne/Popels)

Quantitative PCR (Live Sample -DNA) Variable with physiological state of target cell

ACNS (Hartsig)Polyclonal Antibody & Epifluorescence Microscopy High cross reactivity with other protists& bacteria2001

MarylandDelawareAnalysis Methodfor Natural Water SamplesYear

Analysis Method: Aureococcus Determination


0.E+00

2.E+07

4.E+07

6.E+07

8.E+07

1.E+08

1.E+08

1.E+08

2.E+08

20 22 24 26 28 30 32

6/10/02, LA-8

6/24/02, LA-8

7/8/02, LA-8

Aureococcus (cells/L) vs. Water Temperature οCDelaware Little Assawoman Bay – multiple years


0.0E+00

5.0E+07

1.0E+08

1.5E+08

2.0E+08

2.5E+08

3.0E+08

12 14 16 18 20 22 24 26 28 30 32

2001

2002

2004

Aureococcus (cells/L) vs. Water Temperature oCMaryland Assawoman Bay


0.E+00

2.E+07

4.E+07

6.E+07

8.E+07

1.E+08

1.E+08

1.E+08

2.E+08

0 4 8 12 16 20 24 28 32

6/10/02, LA-8

6/24/02, LA-8

7/8/02, LA-8

Aureococcus (cells/L) vs Salinity (ppt)Delaware Little Assawoman Bay – multiple years


Aureococcus (cells/L) vs. Salinity (ppt)Maryland Assawoman Bay

0.0E+00

5.0E+07

1.0E+08

1.5E+08

2.0E+08

2.5E+08

3.0E+08

12 14 16 18 20 22 24 26 28 30 32

2001

2002

2004


0.0E+00

5.0E+05

1.0E+06

1.5E+06

2.0E+06

2.5E+06

3.0E+06

3.5E+06

4.0E+06

4.5E+06

5.0E+06

5.5E+06

6.0E+06

6.5E+06

7.0E+06

7.5E+06

4/24 5/1 5/22 6/6 6/11 6/19 6/26 7/11 7/17 7/25 8/6 8/29

DE Miller Creek DE Dirickson Creek DE open water MD Ditch MD Montego Bay MD Greys CreekMD open water

MD Montego Bay

Present PresentPresent

MD

MD

DE DEDE Dirickson Creek

2001 - Aureococcus (cells/L): North to South

Collection Dates NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR

0.0E+00

2.5E+07

5.0E+07

7.5E+07

1.0E+08

1.3E+08

1.5E+08

1.8E+08

2.0E+08

2.3E+08

2.5E+08

2.8E+08

5/17 5/29 6/10 6/12 6/24 6/25 7/8 7/10 7/22 8/6 8/21 9/10 9/25 10/1 10/8

MD Ditch

DE Ditch

MD Montego Bay

MD Greys Creek

MD open water

Maryland Category 3 Brown Tide Bloom Index based on Polyclonal Technique

MD Ditch

MDDE

DE MD

Collection Dates

2002 – Aureococcus (cells/L): North to South


0.0E+00

5.0E+06

1.0E+07

1.5E+07

2.0E+07

2.5E+07

3.0E+07

3.5E+07

4.0E+07

4.5E+07

5.0E+07

4/19 5/3 5/18 5/24 6/29 7/1 7/7 7/13 7/27 8/10 8/25 9/8

MD Ditch

DE Ditch

MD Montego Bay

MD Greys Creek

MD open water

MD open water

present

MD

MD

DEDE

Collection Dates

2004 - Aureococcus (cells/L): North to South


0.0E+00

3.5E+07

7.0E+07

1.1E+08

1.4E+08

1.8E+08

2.1E+08

2.5E+08

2.8E+08June 12

Maryland DNR and Delaware DNREC data

April 24 – July 172001

May 17 – July 82002

April 19 – July 72004

Aureococcus (cells/L) – The Ditch


General Observations: Aureococcus OccurrenceLittle Assawoman Bay and Assawoman Bay

• Cell concentrations: annual variability

• Category 3 Bloom concentrations reported in “The Ditch” on 1 occasion in 3 yr. analysis

• Salinity: 7 - 31 ppt

• Water Temperature: 14 -31 C (18-25 C)

• Cell concentrations: spatial variabilityNOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR

Aureococcus Monitoring in 2006

Maryland Assawoman BayRoutine Monitoring: same sites as previous years

late April to mid July, and late September

Delaware Inland Bays * No Routine Monitoring scheduled as of this date

* Continual monitoring of Aureococcus results from MD Coastal Bays

* Rapid Response Capabilities in place for episodic sampling of Phytoplankton Blooms

* Data sonde with fluorescent probe placed in vicinity of “The Ditch”(funding dependent)


Acknowledgements:Ben Anderson, Terri Cole, Ellen Dickey, Rob Gano, Hassan Mirsajadi, Debbie Sullivan, Jeff Tinsman, Robin Tyler, Judy Denver, John Ewart, Cathy Wazniak, Kuo Wong

DE Contact: [email protected] Contact: [email protected]


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