1. Department of the Environment Utilizing a Rapid Laboratory Technique to Assist Public Health Needs Associated with HABs

2. OVERVIEW Cyanobacteria – blue green algae Microcystin – hepatotoxin Enzyme Linked Immuno-Sorbent Assay EVENTS Statistical Analysis –cell counts & ppb

3. ELISA TECHNOLOGY Abraxis, LLC – microtiter well system Delaware, Nebraska DEQ, USGS

4. ELISA TECHNOLOGY Polyclonal AB – allows congener detection of MC and nodularins. Sensitivity Limit: 0.1 ppb Assay range: 0.15 – 5.0 ppb

5. Indirect competitive ELISA (Coated) Analogue Environmental Sample + Poly AB Enzyme (HRP) + substrate (TMB) = color development

6. ELISA TECHNOLOGY Read spectro-photometrically Compared to standard curve, concentration inverse to color intensity

7. 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

8. 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

9. Events 15, 17 samples exceed 10 ppb - recreational (WHO) 1 (‘09) exceed 1 ppb - drinking water guideline

10. 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 1 2 3 4 7 8 6 5

11. 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

12. Transquaking River, Dorchester County

13. 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

15. Sassafras River

16. 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

17. 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

18. Lake Needwood, Mont.Co. Nov ‘09, Woronichinia naegiliana –500,000 –48 ppb July ’10, M. aeruginosa –36 million cell count –3500 ppb

19. Lake Williston- Mill Creek, Caroline Co. M. aeruginosa Aug ’10 –9 million count –370 ppb Sept ‘10 –1.3 million count –56 ppb

20. Developing Bloom Threshold for Management Consideration 2009 - 2010

21. Developing Bloom Threshold for Management Consideration 2009

22. Developing Bloom Threshold for Management Consideration 2009

23. Developing Bloom Threshold for Management Consideration Lake Lariat, 12K TRQ – HMP, 38K

24. 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

25. Inter-agency Cooperation ELISA decreased delays in management decision to protect public health

26. 1800 Washington Boulevard | Baltimore, MD 21230-1718 410-537-3000 | TTY Users: 1-800-735-2258 www.mde.state.md.us 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