Utilizing a Rapid Laboratory Technique to Assist Public Health Needs Associated with HABs

Department of the Environment

Utilizing a Rapid Laboratory Technique to

Assist Public Health Needs Associated with HABs

OVERVIEW

• Cyanobacteria – blue green algae

• Microcystin – hepatotoxin

• Enzyme Linked Immuno-Sorbent Assay

• EVENTS

• Statistical Analysis – cell counts & ppb

ELISA TECHNOLOGY

• Abraxis, LLC – microtiter well system

• Delaware, Nebraska DEQ, USGS

ELISA TECHNOLOGY

• Polyclonal AB – allows congener detection of MC and nodularins.

• Sensitivity Limit: 0.1 ppb

• Assay range: 0.15 – 5.0 ppb

Indirect competitive ELISA

• (Coated) Analogue

• Environmental Sample + Poly AB

• Enzyme (HRP) + substrate (TMB) = color development

ELISA TECHNOLOGY

• Read spectro-photometrically

• Compared to standard curve, concentration inverse to color intensity

Microcystin• Fresh water - not found recently in tidal

areas

• Microcystin does not bioaccumulate

• MDE and DNR program:

– Monitor Dinophysis/Pseudonitschia

– Domoic Acid ELIZA

– No increases in concentration

Events

• 2009, 8 events, 46 samples

• 2010, 9 events, 40 samples

– March, Fountain Rock Quarry, Frederick

– Summer/Fall, Transquaking River, Dorchester

– Nov/July, Lake Needwood, Montgomery County

Events

• 15, 17 samples exceed 10 ppb -recreational (WHO)

• 1 (‘09) exceed 1 ppb - drinking water guideline

Events2009 Blue Green Algae Bloom Sites in Maryland

1.Broadford Lake, Garrett2.Savage Reservoir, Garrett

3.Piney Run Reservoir, Carroll4.Lakes Frank and Needwood, Mont.

5.Trinity Lake, Charles6.Hampshire Lake, Charles

7.Lake Lariat, Calvert8.Transquaking River, Dorchester

1 23

4

7

86

5

Events2009 Sites2010 Sites

1. Fountain Rock Quarry, Fred.

2. Sassafras River, Cecil

3. Lake Needwood, Mont.

4. Trinity Lake, Charles

5. Lake Lariat, Calvert

6.Transquaking, Dorchester

7. Smithville Lake, Caroline

8. Williston Lake, Caroline

9. Bush River, Harford

2

Transquaking River, Dorchester County

Transquaking River• Significant HAB

• Elevated counts and toxins

• Advisory and monitoring

• Remained elevated into the fall

• (‘09) – 20,700 ppb, 118 million count

• (‘10) – 204 ppb, 87,000 count

Sassafras River

Sassafras River

• Historically, M. aeruginosa detected

• This year, M. botrys, non-toxin producer

• (6/18) 90,000 count, 3.2 ppb

• (7/7) 69,000 count,1.5 ppb

• (8/12) 20,000 count, 0.4 ppb

Fountain Rock Quarry• March 2010

• Fountain Rock Quarry

• Plankothrix rubescens

• 5.6 mil cells/ml 500 ppb MC

• June, 300k, 108 ppb

•KMnO4

•Aug, 16 cells, 0.8ppb

Lake Needwood, Mont.Co.

• Nov ‘09, Woronichinia naegiliana

– 500,000

– 48 ppb

• July ’10, M. aeruginosa

– 36 million cell count

– 3500 ppb

Lake Williston- Mill Creek, Caroline Co.

• M. aeruginosa

• Aug ’10

– 9 million count

– 370 ppb

• Sept ‘10

– 1.3 million count

– 56 ppb

Developing Bloom Threshold for Management Consideration

Microcystis sp. v. Microcystin concentration (ppb)

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MC ELIZA ppb

M. a

eru

g (

cell/

ml)

2009 - 2010

Developing Bloom Threshold for Management Consideration

y = 5659.5x - 281991R² = 0.9938

-20,000,000

0

20,000,000

40,000,000

60,000,000

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0.00 5000.00 10000.00 15000.00 20000.00 25000.00

2009

Developing Bloom Threshold for Management Consideration

y = 5697.1x + 29694R² = 0.4522

0

50,000

100,000

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200,000

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0.0 10.0 20.0 30.0 40.0 50.0 60.0

2009

Developing Bloom Threshold for Management Consideration

Lake Lariat, 12K

TRQ – HMP, 38K

Developing Bloom Threshold for Management Consideration

• 2004, Dyble, found that:

• Microcystis cell densities only explained 34% of the variation in total microcystin concentration

• Several factors likely contribute to this poor correlation,

• (1) the inclusion of both toxic and nontoxic strains of Microcystis in cell counts,

• (2) variability in toxin production within a toxic strain related to cell growth, and

• (3) changing proportions of toxic to nontoxic genotypes in bloom populations

• Microcystin Concentrations and Genetic Diversity of Microcystis in the Lower Great Lakes

• Julianne Dyble, et al, NOAA, Great Lakes Environmental Research Laboratory, 2205 Commonwealth Blvd, 2004

Inter-agency Cooperation• ELISA decreased delays in management

decision to protect public health

1800 Washington Boulevard | Baltimore, MD 21230-1718410-537-3000 | TTY Users: 1-800-735-2258

www.mde.state.md.us

Maryland Department of the Environment

Water Quality Restoration and PreservationProgram

Charles PoukishChris Luckett

Nick KaltenbachJeff Carter

Emily Anderson, Kevin KellyPatricia Brady

Thanks to Cathy Wazniak, Celia Dawson,Walt Butler, DNR

Cliff Mitchell, Nancy Servatius, DHMH