Harmful Algal Bloom Research at Stone Lab: Monitoring Blooms and Determining. Drivers of Bloom Toxicity

Harmful Algal Bloom Research at Stone Lab: Monitoring Blooms and Determining WMAO/OWRC Luncheon October 12, 2016 Drivers of Bloom Toxicity Justin Chaffin, PhD Stone Laboratory, Ohio Sea Grant Chaffin.46@osu.edu

Most algae are not bad for lakes Algae are tiny plant-like organisms 50% of Earth s oxygen produced by algae Every other breath you take, thank algae Base of the lake food web Good Algae are food for zooplankton Zooplankton are food for small fish Large fish Algae Food Zooplankton (small shrimp-like creatures) Small fish Oxygen

There are hundreds of species of algae in lakes Algae need: Sunlight, fertilizers, carbon dioxide

Lake Erie produces the most fish of all the Great Lakes because it has the most algae Highest nutrient concentrations Warmest water temperature Lake Erie 50% of fish 2% of the water Lake Superior 2% of fish 50% of water

Too much of the wrong kind of algae is harmful Harmful = Has the potential to produce toxins. Harmful impacts on ecosystems Algal = Blue-green algae (cyanobacteria) Red tide (dinoflagellates) Bloom = Biomass that far exceed normal geology.com Hungeree.com

Lake Erie harmful algal blooms Lake Erie blooms are a type of Cyanobacteria called Microcystis Also called blue-green algae Technically true bacteria Require sunlight like algae Microcystis produces the toxin microcystin that was responsible for Toledo s 2014 water crisis. Other HABs in Lake Erie Sandusky Bay Central Basin J.Chaffin

Tracking blooms requires multiple scales of monitoring Blooms visible (and quantifiable) from space NOAA HAB bulletin Entire lake once or twice a week But: No information on toxicity, nutrients, species Water samples from boats A lot of information on toxicity, nutrients, species, molecular But: Few locations and low frequency. Lab analysis time lag Buoys equipped with sensors Real-time data. Early warning of a bloom Potential issues: Infrequent calibration, Zebra mussel clogging, Variable fluorescence per chlorophyll. Do sensors work?

Charter boat captains monitor western basin water quality HABs hurt charter captain business. 30% less business in 2015 Trips cancelled due to bloom Customers not returning In 2012 (after large 2011 bloom), captains asked How can we help? Funded by Ohio EPA Surface water in 2014 Environmental Education in 2013, 2015, 2016

Charter boat captains sampling kit

Charter boat captains monitor western basin water quality Captains record: Lat/Long, Depth, Surface temp Measure Secchi Disk depth Sample surface water with 0-2m intergraded tube sampler Drop off cooler at designated bait shops/marinas. 7 samples per week Captains receive data updates each week. Answer How is the lake doing this year? Basspro.com Eriehopper.com

Charter boat sampling data update Total algae levels have remained lower in September and October than August. Highest algae levels were recorded in early May during a diatom bloom. Microcystin levels slightly increased in late September and October. This increase was associated with an increase of cyanobacteria (a.k.a. blue-green algae). 2016 overall has had much less algae and microcystin than previous years.

Charter boat sampling data update Phosphorus is the main nutrient that limits algae growth. Phosphorus levels were greater in August and September than in May, June, and July. Recent phosphorus levels have been similar to levels measured in previous years. The phosphorus target for the western basin is 15 ppb. During late May and early June the western basin was below that target level. Nitrogen is the second most important nutrient for algae. Nitrate is the most common form on nitrogen in Lake Erie. Nitrate levels have steadily decreased since very high levels were measured in April.

The map shows where the captains have collected samples by month during 2016. There has been a good coverage of the entire western basin. Captains have collected 145 samples as of October 3.

Data buoys provide real-time data Fixed monitoring stations Records biological, physical, chemical, and meteorological variables Continuous, real-time data Data can be utilized by lake managers, researchers, water treatment plant operators, tourists and the general public Gibraltar Buoy

HABs Real-Time Monitoring Slide courtesy of Ed Verhamme

Data buoys measure water quality 0.7 meter from the surface. Buoy parameters (15 min) Dissolved Oxygen Temperature ph Specific Conductivity Water clarity (NTU) Total algae (chlorophyll fluorescence) Cyanobacteria (phycocyanin fluorescence) Lake bottom

Potential issues with the sensors? Infrequent calibration Sensor drift throughout deployment? Algae biomass is not measured Sensors measure fluorescence, which can change with physiological status of the cell Clogging from Dreissena mussels Low water exchange

Water samples collected next to the buoy Deployed May October in 2015 and 2016 Water samples next to the buoy throughout summer using 0-2 meter integrated tube sampler (several samples /week) Microcystin (total and extracellular) Chlorophyll and phycocyanin Phytoplankton biovolume data FluoroProbe (chlorophyll a associated with green algae, diatoms, and cyanobacteria) Nitrate, ammonium, TKN, TP, DRP, Si TSS & NVSS Secchi disk depth 125 samples

Water Sample BUOY OHIO SEA GRANT AND STONE LABORATORY 2015 large bloom at Stone Lab buoy July 24, 2015 Chaffin unpublished

Water Sample BUOY OHIO SEA GRANT AND STONE LABORATORY 2016 bloom just above detection limit Chaffin unpublished

Water Sample BUOY OHIO SEA GRANT AND STONE LABORATORY The 2016 bloom never reached the islands. NOAA HAB Bulletin August 11 2016

Good relationship between buoy BGA-PC RFU and water sample cyanobacteria Chaffin unpublished

Total Chlorophyll: Different trends between the two years and low correlation Chaffin unpublished

Buoy NTU sensor good indicator of water clarity. Chaffin unpublished

Microcystin concentration tracked with buoy data Chaffin unpublished

Can buoy sensor data be used to predict water quality? Cyanobacteria: Yes. R 2 = 0.96 Total chlorophyll: No. R 2 <0.50; different relationship between the years Water clarity: Yes. R 2 = 0.86 for TSS Yes. R 2 = 0.81 for Secchi Disk Microcystin concentration tracked the buoy cyanobacteria fluorescence data.

Cyanobacterial blooms just not western basin problem. Microcystis in western basin A lot of research ongoing Many research groups Planktothrix in Sandusky Bay See: Davis et al. 2016 ES&T Dolichospermum (Anabaena) in central basin Research focus on hypoxia Little known about phytoplankton NOAA HAB bulletin, July 15 2015

Dolichospermum in the central basin Formally called Anabaena Nitrogen fixing cyanobacterium Typically found in low N and high P waters. Not believed to be a microcystin producer in Lake Erie (Rinta-Kanto et al. 2009) Early July blooms 4 of last 5 years D. lemmermannii D. circinale D. planctonicum

Central basin Dolichospermum bloom occurred in high nitrate and high TN:TP ratios. BLOOM DRP was below detection in all samples. A N-fixer HAB in high N waters could indicate that Fe was limiting growth. Chaffin unpublished 2013 bloom: NOAA July 7, 2016: Brian Fowler, Lake Co Metroparks

Role of nitrogen in bloom growth and toxicity Human activities are increasing the amount of nitrogen available to ecosystems. Gilbert et al. 2014

Research question: How will the increase use of nitrogen affect cyanobacterial blooms in terms of growth and toxicity? Experiment addressed: Forms of nitrogen Nitrate, Ammonium, Urea Loading rate Single high: Storm pulsed Low sustained: Internal recycling Light level Interaction with N?

Nitrogen limits OHIO SEA GRANT Lake AND STONE Erie LABORATORY cyanobacteria abundance in August and September Control +P +N +P&N See: Chaffin, Bridgeman, Bade. 2013. Nitrogen constrains the growth of late summer cyanobacterial blooms in Lake Erie. Advances in Microbiology. 6: 16-26

Nitrogen constrains Planktothrix blooms in Sandusky Bay Davis et al. 2015. Environ. Sci. Technol. 49: 7197 7207

Is nitrogen important for cyanobacteria toxin production? Non-nitrogen fixing cyanobacteria dominate the toxic blooms in Lake Erie Microcystis in the main lake Planktothrix near river mouths and Sandusky Bay Microcystin-LR is 14% nitrogen by mass. Microcystis is only 7% nitrogen by mass (Chaffin et al. 2011). Thus, making microcystin is expensive in terms of nitrogen. Microcystis and Planktothrix cannot grow or make microcystin without combined nitrogen. Nitrate, Ammonium, Urea

Combined nitrogen enters a lake through rivers and by recycling in sediments and water column NO3, NH4, Organic N Particulate N Following storms, near shore nitrate concentrations in Lake Erie exceed 100 µmol/l (Chaffin et al. 2013) Ammonium is recycled at a rate of 2.5 µmol/l/h in Taihu (Paerl et al. 2011) NH4 and Urea NO3 NH4 and Urea Sediment Nitrogen

Three study locations. OHIO SEA GRANT AND STONE LABORATORY Sandusky Bay Planktothrix agardhii Maumee Bay Microcystis aeruginosa Bass Islands Microcystis aeruginosa

Three study locations. OHIO SEA GRANT AND STONE LABORATORY Data shown in today s presentation is from Maumee Bay experiments Maumee Bay August 6 2014 Microcystis aeruginosa [chla] = 68 µg/l

Nitrogen form X loading rate experiments. Control: No additional nutrients Phosphorus: 1 µmol/l added at beginning Single high Nitrate, P: Single high Ammonium, P: 100 µmol N/L added at beginning Single high Urea-N,P: Low continuous Nitrate, P: Low continuous Ammonium, P: 8.3 µmol N/L added every 4 hours Low continuous Urea-N, P: The single high enrichments simulate nitrogen concentration following a rain water pulse from a river Low continuous enrichments simulate nitrogen recycling rate of 2.07 µmol/l/h. The cumulative nitrogen added in the low continuous treatments equaled that of the single high enrichments. 3 separate replicates per treatment

Experiment methods Add N and P every 4 hours

Experiment methods OHIO SEA GRANT AND STONE LABORATORY

Measurements from bottles OHIO SEA GRANT AND STONE LABORATORY Microcystin Hour 0, 48 Total and extracellular Chlorophyll Hour 0, 48 Dissolved nutrients Hour 0, 48 Nitrate, ammonium, urea, phosphate Phytoplankton biovolume Hour 0, 48 Gene expression Hour 0, 4, and 48 mcyd RPOC1 (RNA polymerase gamma subunit) The 4 hour sample will show how quick genes are up- or downregulated

Nitrogen enrichment increased chlorophyll, and highest chlorophyll seen with ammonium enrichments Chaffin unpublished Gray bars t = 48 h Bars = mean (± 1 standard error) of 3 replicates. Letters = Tukey test results, where mean of A is greater than mean of B

Nitrogen increased microcystin concentration and highest microcystin with increased nitrate and urea Chaffin unpublished Gray bars t = 48 h Bars = mean (± 1 standard error) of 3 replicates. Letters = Tukey test results, where mean of A is greater than mean of B

0.18 OHIO SEA GRANT AND STONE LABORATORY 0.16 0.14 0.12 0.10 0.08 0.06 0.04 0.02 - t = 0 t = 4, Control t = 4, P only t = 4, Nitrate (once) t = 4, Ammonium (once) t = 4, Urea (once) t = 4, Nitrate (every 4h) t = 4, Ammonium (every 4h) t = 4, Urea (every 4h) t = 48, Control Relative expression (mcyd:rpoc1) Microcystin synthesis genes up-regulated within 4 hours of exposure to 100 µm ammonium and urea* but not nitrate. A A B B A B B B t = 4 h Bars = Mean ± SE; A and B = significantly up-regulated (p < 0.05) t = 48, P only t = 48, Nitrate (once) t = 48, Ammonium (once) t = 48, Urea (once) t = 48, Nitrate (every 4h) t = 48, Ammonium (every 4h) t = 48, Urea (every 4h) Chaffin unpublished

0.18 0.16 0.14 0.12 0.10 0.08 0.06 OHIO SEA GRANT AND STONE LABORATORY 0.04 0.02 - t = 0 t = 4, Control t = 4, P only t = 4, Nitrate (once) t = 4, Ammonium (once) t = 4, Urea (once) t = 4, Nitrate (every 4h) Relative expression (mcyd:rpoc1) Microcystin synthesis genes expression was similar between 100 µm and 8.3 µm nitrogen 4 hours after exposure 100 µm 8.3 µm A A Small and large pulses of reduced nitrogen increased B microcystin synthesis gene expression. A B B B B Bars = Mean ± SE; A and B = significantly up-regulated (p < 0.05) t = 4, Ammonium (every 4h) t = 4, Urea (every 4h) t = 48, Control t = 48, P only t = 48, Nitrate (once) t = 48, Ammonium (once) t = 48, Urea (once) t = 48, Nitrate (every 4h) t = 48, Ammonium (every 4h) t = 48, Urea (every 4h) Chaffin unpublished

0.18 0.16 0.14 0.12 0.10 0.08 0.06 0.04 0.02 - t = 0 t = 4, Control t = 4, P only t = 4, Nitrate (once) t = 4, Ammonium (once) t = 4, Urea (once) t = 4, Nitrate (every 4h) t = 4, Ammonium (every 4h) t = 4, Urea (every 4h) t = 48, Control Relative expression (mcyd:rpoc1) mcy expression maintained 48 hours after N exposure. A A B B A B B B Bars = Mean ± SE; A and B = significantly up-regulated (p < 0.05) t = 48, P only t = 48, Nitrate (once) t = 48, Ammonium (once) t = 48, Urea (once) t = 48, Nitrate (every 4h) t = 48, Ammonium (every 4h) t = 48, Urea (every 4h) Chaffin unpublished

0.18 OHIO SEA GRANT AND STONE LABORATORY 0.16 0.14 0.12 0.10 0.08 0.06 0.04 0.02 - t = 0 t = 4, Control t = 4, P only t = 4, Nitrate (once) t = 4, Ammonium (once) t = 4, Urea (once) t = 4, Nitrate (every 4h) t = 4, Ammonium (every 4h) t = 4, Urea (every 4h) Relative expression (mcyd:rpoc1) After 48 hours, nitrate and P-only increased expression. No difference between single dose and every 4 hour doses. A A B B A B B B Bars = Mean ± SE; A and B = significantly up-regulated (p < 0.05) t = 48, Control t = 48, P only t = 48, Nitrate (once) t = 48, Ammonium (once) t = 48, Urea (once) t = 48, Nitrate (every 4h) t = 48, Ammonium (every 4h) t = 48, Urea (every 4h) Chaffin unpublished

Overall summary Charter captains augment existing water quality monitoring programs Citizen science. Public education Data buoys can be used to predict cyanobacteria and water clarity. But not total chlorophyll Dolichospermum (Anabaena) blooms in the central basin despite high N:P ratios Blooms 4 of last 5 summers Nitrogen enrichment stimulates microcystin production by Microcystis and Planktothrix. Low pulses and large pulses triggered same responses. Does not down play the role of phosphorus

Acknowledgements Ohio EPA (Charter captain sampling) Ohio Water Resources Center (Buoy project) Ohio Department Higher Education (Central Basin) Ohio Sea Grant (Nitrogen and toxicity) Collaborators: The Captains, Tim Davis, Doug Kane, Darren Bade, George Bullerjahn, Mike McKay, Rick Stumpf, Heather Raymond Fondriest Environmental Stone Lab REU students and Friends of Stone Lab Stone Lab staff

Questions & Discussion MODIS. October 2011