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

Events 2009 Blue Green Algae Bloom Sites in Maryland 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

Events 2009 Sites 2010 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

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 KMnO 4 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

Developing Bloom Threshold for Management Consideration 2009

Developing Bloom Threshold for Management Consideration 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 | TTY Users: Maryland Department of the Environment Water Quality Restoration and Preservation Program Charles Poukish Chris Luckett Nick Kaltenbach Jeff Carter Emily Anderson, Kevin Kelly Patricia Brady Thanks to Cathy Wazniak, Celia Dawson, Walt Butler, DNR Cliff Mitchell, Nancy Servatius, DHMH