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
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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)
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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 %
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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)
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR
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]
NOT FOR REPRODUCTION WITHOUT CONSENT OF AUTHOR