utilizing a rapid laboratory technique to assist public health needs associated with habs
DESCRIPTION
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. - PowerPoint PPT PresentationTRANSCRIPT
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)
1
10
100
1,000
10,000
100,000
1,000,000
10,000,000
100,000,000
1,000,000,000
1 10 100 1000 10000 100000
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
80,000,000
100,000,000
120,000,000
140,000,000
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
150,000
200,000
250,000
300,000
350,000
400,000
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