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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.
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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
2009 Blue Green Algae Bloom Sites in Maryland 2 Events 1 3 4 1.Broadford Lake, Garrett 2.Savage Reservoir, Garrett 3.Piney Run Reservoir, Carroll 4.Lakes Frank and Needwood, Mont. 5.Trinity Lake, Charles 6.Hampshire Lake, Charles 7.Lake Lariat, Calvert 8.Transquaking River, Dorchester 5 8 6 7
2009 Sites 2010 Sites Events 2 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
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 • 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
March 2010 • Fountain Rock Quarry • Plankothrixrubescens • 5.6 mil cells/ml • 500 ppb MC • June, 300k, 108 ppb • KMnO4 • Aug, 16 cells, 0.8ppb Fountain Rock Quarry
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
2009 - 2010 Developing Bloom Threshold for Management Consideration
2009 Developing Bloom Threshold for Management Consideration
2009 Developing Bloom Threshold for Management Consideration
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
Maryland Department of the Environment Water Quality Restoration and PreservationProgramCharles PoukishChris LuckettNick KaltenbachJeff CarterEmily Anderson, Kevin KellyPatricia BradyThanks to Cathy Wazniak, Celia Dawson, Walt Butler, DNRCliff Mitchell, Nancy Servatius, DHMH 1800 Washington Boulevard | Baltimore, MD 21230-1718410-537-3000 | TTY Users: 1-800-735-2258www.mde.state.md.us