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Ohio Water Microbiology Lab

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OWML: Completed Projects

Project Title: Using new tools to better understand and predict harmful cyanobacterial algal blooms (cyanoHABs) at Ohio Lake Erie and inland beaches.

Project chief:  Donna Francy

Project support:  Erin Stelzer, Chris Ecker, Carrie Huitger, Amie Brady

Project funding: Ohio Water Development Authority, U.S. Geological Survey

Cooperators:  University of Toledo, Clermont County Soil and Water Conservation District, Ohio Department of Natural Resources, U.S. Environmental Protection Agency, Ohio Environmental Protection Agency, U.S. Army Corps of Engineers

Project duration:  2013-2015

Introduction and problem:
In Ohio, local health officials have identified the presence of cyanobacterial harmful algal blooms (cyanoHABs) during the summer and early fall seasons at some Lake Erie and inland lake beaches and access areas. The cyanoHABs are caused by cyanobacteria (blue-green algae) that produce microcystin and other toxins.

Microcystins are commonly produced by cyanobacteria in the genera Microcystis, Planktothrix, and Dolichospermum (formerly Anabaena) (Rantala et al., 2006). For toxin production to occur, the microcystin synthetase genes (mcy) must be present in the genome of toxic strains. Known microcystin-producing genera include both toxic strains (with the mcy genes) and nontoxic strains (without the mcy genes), which can be differentiated only by molecular detection methods such as quantitative polymerase chain reaction (qPCR). Whereas DNA-based qPCR methods reveal the presence of toxin genes (irrespective of whether they are actively producing toxin), RNA-based methods can detect microcystin-producing cyanobacteria that are actively expressing the toxin genes (Sipari et al., 2010).

Optical sensors that measure algal pigments have been used to provide early warnings of cyanobacterial presence in recreational waters (Marion et al., 2012) and drinking-water sources (Brient et al., 2008; McQuaid et al., 2011). These include sensors based on the fluorescent properties of chlorophyll (indicative of total phytoplankton abundance) and phycocyanin (indicative of cyanobacterial abundance).

Current methods for identifying cyanobacteria by microscopy are time consuming and are unable to identify whether a strain has the ability to produce toxins. Methods are needed to better identify and understand the potential for toxin production in cyanobacterial populations and, at the same time, provide an early warning system for toxin production. 

Goals and objectives:
The overall goal is to apply qPCR methods and optical sensor tools to better understand the link between cyanobacteria type and abundance, environmental and water-quality factors, and occurrence of toxins. Objectives are to

  1. understand how the cyanobacterial community progresses from nontoxic to toxic strains and the progression from unexpressed to expressed genes;
  2. determine the relations between toxin production and environmental and water-quality variables to help support predictive capabilities for cyanoHABs; and
  3. determine whether adding new tools to daily monitoring programs at beaches could be used to provide better, more relevant public health information to beach managers and the public as compared to the use of currently used methods for determining toxin concentrations and (or) phytoplankton abundance.

Approach:
Sampling sites and frequency
Samples were collected at Ohio recreational sites (Fig. 1) from May–November in 2013 and 2014 using procedures described in Graham and others (2008).

During 2013, samples were collected approximately monthly from the four Lake Erie and four inland lake beach sites listed below. Samples were collected to facilitate an initial assessment and select sites for more intensive sampling during 2014.

  1. Maumee Bay State Park Lake Erie beach
  2. Maumee Bay State Park inland lake beach (control site)
  3. Sandusky Bay at Bay View
  4. Lake Erie at Lakeview Park, Port Clinton
  5. Buck Creek State Park beach (CJ Brown Reservoir)
  6. East Fork State Park Campers beach (Harsha Lake)
  7. Deer Creek State Park beach
  8. Buckeye Lake Crystal beach

During 2014, samples were collected approximately weekly at five sites at three lakes.

  1. Maumee Bay State Park Lake Erie beach
  2. East Fork State Park Main beach
  3. East Fork State Park Campers Beach
  4. Buckeye Lake Fairfield beach
  5. Buckeye Lake Onion Island

Constituents measured included the following:

  • Specific conductance, dissolved oxygen, pH, temperature, chlorophyll, and phycocyanin using an in-situ water-quality sensor at the time of sampling
  • Secchi depth at the time of sampling
  • Samples preserved on ice and shipped to laboratories for
    • Dissolved and total nutrients (USGS National Water Quality Laboratory)
    • Cyanotoxins (microcystin, saxitoxin, and cylindrospermospsins by ELISA (USGS Kansas Organic Geochemistry Laboratory)
    • Phytoplankton abundance and community structure by microscopy (cell counts)
    • Cyanobacteria genes by qPCR on four levels (USGS Ohio Water Microbiology Laboratory)
      • Total cyanobacteria (Rinta-Kanto et al., 2005)
      • Total Microcystis (Rinta-Kanto et al., 2005)
      • Genus-specific DNA mcyE assays for Microsystis and and Planktothrix (Rantala et al., 2006)
      • Genus-specific RNA mcyE assays for expressed genes (Sipari et al., 2010)

Results and Conclusions:
Data are available at the USGS National Water Information System web site (NWIS-web) by site identification number (http://waterdata.usgs.gov/oh/nwis/qw/). The final project report (Francy and others, 2015) is available on-line (http://dx.doi.org/10.3133/sir20155120).

Statistically significant correlations were found between microcystin concentrations and phycocyanin, turbidity, pH, streamflow, lake level change, nutrient concentrations, cyanobacterial genes, and cyanobacterial cell counts. The relations between these factors and microcystin concentrations were site specific and even differed at the same site depending on whether data were continuously or discretely measured. Models were developed at Maumee Bay State Park beach for predicting microcystin concentrations above or below the Ohio Recreational Public Health Advisory level of 6 ug L-1 (Ohio Environmental Protection Agency, 2015). The results of this study showed that environmental factors are promising for use in site-specific models for cyanoHABs at freshwater lakes. Future studies should focus on collecting more frequent data on several consecutive days each week before, during, and after the cyanoHAB season to develop robust site-specific models.

References:
Brient, L., Lengronne, M., Bertrand, E., Rolland, D., Sipel, A., Steinmann, D., Baudin, I., Legeas, M., Le Rouzic, B., and Bormans, M., 2008, A phycocyanin probe as a tool for monitoring cyanobacteria in freshwater bodies: J. Environmental Monitoring 10(2), 248–255.

Francy, D.S., Graham, J.L., Stelzer, E.A., Ecker, C.D., Brady, A.M.G., Struffolino, P., and Loftin, K.A., 2015, Water quality, cyanobacteria, and environmental factors and their relations to microcystin concentrations for use in predictive models at Ohio Lake Erie and inland lake recreational sites, 2013–14: U.S. Geological Survey Scientific Investigations Report 2015–5120, 58 p., http://dx.doi.org/10.3133/sir20155120.

Graham, J.L., Loftin, K.A., Ziegler, A.C., and Meyer, M.T., 2008, Guidelines for design and sampling for cyanobacterial toxin and taste-and-odor studies in lakes and reservoirs: U.S. Geological Survey Scientific Investigations Report 2008–5038, 39 p.

McQuaid, N., Zamyadi, A., Prevost, M., Bird, D.F., and Dorner, S., 2011, Use of in vivo phycocyanin fluorescence to monitor potential microgystin-producing cyanobacterial biovolume in a drinking water source: J. Environmental Monitoring 13, 455–463.

Marion, J.W., Lee, J., Wilkins, J.R. III, Lemeshow, S., Lee, C., Waletzko, E.J., and Buckley, T.J., 2012, In vivo phycocyanin fluorometry as a potential rapid screening tool for predicting elevated microcystin concentrations at eutrophic lakes: Environ. Sci. Technol. 46, 4523–4531.

Ohio Environmental Protection Agency, 2015, State of Ohio—harmful algal bloom response strategy for recreational waters, accessed October 2015 at http://www.epa.state.oh.us/dsw/HAB.aspx

Rantala, A., Rajaniemi-Wacklin, P., Lyra, C., Lepisto, L., Rintala, J., Mankiewicz-Boczek, J., and Sivonen, K., 2006, Detection of microcystin-producing cyanobacteria in Finnish lakes with genus-specific microcystin synthetase gene E (mcyE) PCR and associations with environmental factors: Appl. Environ. Microbiol. 72, 6101–6110.

Rinta-Kanto, J.M., Ouellette, A.J.A., Boyer, G.L., Twiss, M.R., Bridgeman, T.B., and Wilhelm, S.W., 2005, Quantification of toxic Microcystis spp. during the 2003 and 2004 blooms in western Lake Erie using quantitative real-time PCR: Environ. Sci. Technol. 39, 4198-4205.

Sipari, H., Rantala-Ylinen, A., Jokela, J., Oksanen, I., and Sivonen, K., 2010, Development of a chip assay and quantitative PCR for detecting microcystin synthetase E gene expression: Appl. Environ. Microbiol. 76, 3797–3805.