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Spatial and Temporal Distribution of Cyanobacterial Toxins in Lake Erie Principal Investigator: Greg Boyer Professor of Biochemistry State University of New York College of Environmental Science and Forestry Syracuse, NY 13210 315-470-6825 (voice) 315-470-6856 (fax) Email: glboyer@esf.edu Proposed Budget: Dates of the Project: Federal funds requested: $0 Matching funds: na May 1, 2005 – April 30, 2006 Executive Summary: Since the mid 1990’s, Lake Erie has experienced widespread but sporadic blooms of toxic cyanobacteria While the most intense blooms have occurred in the more eutrophic western basin, toxic outbreaks have also occurred in Sandusky Harbor and near Long Point in the eastern basin of the lake In general, the causative organism is reported to be Microcystis aeruginosa which can produce the heptatoxic microcystins This is an oversimplification of the problem as other microcystin-producing species, such as Planktothrix and Anabaena, and other cyanobacterial toxins, such as anatoxin-a and cylindrospermopsin, also occur in the lake This project will determine the occurrence of four classes of cyanobacterial toxins in the three basins of Lake Erie and prepare detailed maps on their distribution and change with time This information will be of use to water supply companies, government officials interested in protecting human health, lake modelers, and other lake scientists studying food webs and the planktonic ecology of the lake Principle funding for this project will come from NOAA’s Lower Great Lakes MERHAB project This request is for ship time only G.L Boyer page 2 Scientific Rationale In 1995, Lake Erie experienced a large bloom of toxic cyanobacteria in the western basin This bloom was caused by Microcystis aeruginosa which produced the hepatotoxin microcystinLR and lesser amounts of the demethyl (Asp3) microcystin-LR and microcystin-AR (Brittain et al, 2000) Since that initial description, toxic Microcystis has continually reoccurred in the western basin of Lake Erie with toxin levels often exceeding the WHO advisory level of µg L-1 (Table 1) The MERHAB – Lower Great Lakes project (MERHAB-LGL) was started in 2002 to investigate the spatial distribution and chemical diversity of cyanobacterial toxins and species in the lower great lakes (Lakes Ontario, Erie and Champlain) The goal of MERHAB is to provide monitoring and event response strategies for harmful algal blooms suitable for use by end users such as health departments, state departments of environmental conservation and water supply providers As part of that project, we have sporadically sampled Lake Erie on a number of different cruises of opportunity Most of these cruises have been in collaboration with the MELEE (Microbial Ecology of the Lake Erie Ecosystem) working group, but we have also obtained samples from other researchers The results from these surveys are shown below Table 1: Recent Occurrence and Distribution of Cyanobacterial Toxins in Lake Erie Cruise and Date # samples % containing Highest Comments toxin measured value * Brittain et al 44 MC’s ~10% 3.4 µg L-1 MC Western basin only Sept 1996 MELEE-VII MC’s % ATX 14% PSP’s % 0.7 µg L-1 MC 0.04 µg L-1 ATX Whole lake survey with highest values at Sandusky, Long Point and Rondeau Bays July 2002 119 MELEE-VIII 59 MC’s 41% ATX 5% 0.65 µg L-1 MC 0.11 µg L-1 ATX Whole-lake survey with highest values in the western basin and Sandusky Bay 48 MC’s 60% ATX 4% 21 µg L-1 MC 0.2 µg L-1 ATX Western basin only, Highest values were obtained near the Maumee River 40 MC’s 38% ATX 33% CYL 0% >1 µg L-1 MC 0.6 µg L-1 ATX Highest values near the Maumee River and in Sandusky Bay July 2003 Lake Guardian & OSU, August 2003 MELEE-IX July 2004 MC’s 85% 2.4 µg L-1 MC Western basin only -1 ATX 31% 0.07 µg L ATX August 2004 CYL 15% 0.18 µg L-1 CYL *These results are extrapolated from Figure in Brittain et al (2000) Abbreviations: MC’s = microcystins, ATX = anatoxin-a, PSP = saxitoxin + neosaxitoxin, CYL = cylindrospermopsin RV Limnos 13 G.L Boyer page Several conclusions are apparent from this table First, both the overall level and spread of microcystin toxicity in the lake appears to be gradually increasing from the original report by Brittain et al They reported two stations in 1996 that exceeded the WHO advisory threshold of 1.0 µg L-1, whereas in 2003 and 2004, a number of stations exceeded that threshold with the highest microcystin concentration exceeding 20 µg L-1 Microcystins have also been found outside the western basin in places such as Long Point Bay (eastern basin), Rondeau Bay (central basin) and Sandusky Harbor (Ouellette et al 2005) We now suspect that organisms other than Microcystis may be producing significant amounts of microcystins and that simply counting Microcystis colonies in the water column may not give an accurate measure of the potential risk Preliminary results also suggest that other cyanobacterial toxins such as the neurotoxin anatoxina and the hepatotoxin cylindrospermopsin are present in Lake Erie (Table 1, Yang et al., 2005, Boyer, unpublished) Unfortunately the development of detailed distribution maps of seasonal toxicity has not been possible as we have not had access to sufficient ship time to properly canvas the lake Most of these results have been obtained from ships of opportunity focused heavily on the western basin Sample collection protocols have also varied widely between cruises, making it difficult to compare results across years The International Field Year on Lake Erie offers a unique opportunity to prepare these detailed toxin distribution maps on a lake-wide basis Understanding the distribution of cyanobacterial toxins and their change with time is essential for the proper placement of observing stations if they are to protect against harmful algal blooms These results will be useful for water supply providers, government officials interested in protecting human health, lake modelers, and other lake scientists studying food webs and the planktonic ecology of the lake Hypothesis and Questions addressed: Preliminary studies suggest that the distribution of cyanobacterial toxins in Lake Erie extends beyond the occurrence of microcystins in the western basin Our working hypothesis is that the occurrence of microcystins in the eastern and central basins will represent distinct bloom events, and that any toxicity measured in the water column is not a result of transport from the western basin Furthermore we hypothesize that anatoxin-a producing species exist in Lake Erie and that this neurotoxin represents an unknown risk to recreational and drinking water users of the lake One of the reasons we think this toxin is poorly reported is that it is relatively unstable in the water column and we not have sufficient sampling density to map its distribution Project Objectives: The objectives of this proposal are three-fold (1) To determine the spatial distribution of particulate cyanobacterial toxins in Lake Erie (2) To examine the change in that spatial distribution with time over an entire growing season (3) To evaluate the chemical diversity of microcystin(s) produced in situ, especially as it affects the different assays for microcystins Project Approach and Methods: The basic methodology for this project is well established, having been used to collect and analyze more than 2000 samples over the last years as part of the MERHAB-LGL project This protocol was adapted to both minimize the time on station for the ship and provide a sufficiently large sample to determine toxin content in the more oligotrophic areas such as the central basin G.L Boyer page Objectives 1and 2: Spatial and Temporal Distribution: Key to getting an accurate map of the spatial and temporal distribution of cyanobacterial toxins in Lake Erie is to collect a large number of samples over a widely dispersed area at multiple times during the summer Here we are proposing to participate on 5-6 whole lake cruises (one each month) to collect samples for toxin analysis Sample Collection: Briefly, a 23-liter grab sample is collected at a fixed depth using a high speed submersible pump In the past, we have deployed this pump by hand off the side of the RV Limnos This usually requires < minutes total time once the ship is stopped and stabilized The carboy is then returned to the ship’s lab and 20L immediately filtered through a 150 mm filter holder equipped with a 934AH glass fiber filter using a high speed peristaltic pump Separate 1-L samples are filtered for chlorophyll and molecular (DNA) analysis These filters are stored at < -20º until the end of the cruise Pigments (chlorophyll and phycocyanin) are measured on board the ship using Turner Designs fluorometers If necessary, samples will be sterile filtered for dissolved nutrient analysis For most samples, a net tow is collected using a small Wisconsin net for identification of the major cyanobacterial species in the water column Once back on shore, filters for toxin analysis are extracted by sonication with 10 ml of 50% methanol acidified with 1% acetic acid, clarified by centrifugation, and the extract used for analysis of the different toxins Microcystins are measured using a combination of different assays including an enzyme-linked immunoassay or ELISA, inhibition of the protein phosphatase 1A (PPIA: Carmichael and An 1999), and by high performance liquid chromatography (HPLC) coupled with either UV or mass selective (MS) detection (Harada 1996) Anatoxin-a is determined by HPLC after derivatization with 7-fluoro-4-nitro-2, 1, 3benzoxadiazole (NBD-F) (James and Sherlock 1996) and confirmed by LCMS of the free or NBD-derivatized toxin The PSP toxins (saxitoxin, neosaxitoxin, and gonyautoxins 1-4) are measured by HPLC with fluorescent detection after either chemical (PCRS: Oshima 1995) or electrochemical (ECOS: Boyer and Goddard 1999) post-column derivatization Cylindrospermopsin is measured by HPLC using PDA detection and confirmed by LCMS (Li et al., 2001) Control experiments have shown that this extraction protocol recovers >90% of the microcystins, PSP toxins, anatoxin-a, and cylindrospermopsin in the sample Assuming normal concentration factors1, the detection limits for the different toxins using this protocol are: Detection limit for the different toxins expressed in µg per L starting lake water Microcystins: Anatoxin-a PSP toxins Cylindrospermopsin < 0.003 µg L-1 (PPIA), < 0.01 µg L-1 (HPLC-PDA) < 0.001 µg L-1 (HPLC-FD) < 0.1 µg L-1 (HPLC-ECOS) < 0.01 µg L-1 (HPLC-PDA) This project will specifically focus on the distribution of particulate cyanobacterial toxins because the existing methodology to accurately measure dissolved toxins in dilute samples such as Lake Erie has not reached a point of sophistication as to provide high quality results We are currently developing novel approaches for differentiating between dissolved and particulate A normal concentration factor assumes a 20 L sample was filtered and extracted in 10 mL For more eutrophic sites such as the western basin of Lake Erie and the Bay of Quinte in Lake Ontario, we often cannot filter 20 L without plugging the filter In those cases, the detection limit per L lake water is higher, but we also have sufficient biomass to easily measure the toxin content of the water if present and can express it on a per chlorophyll basis G.L Boyer page microcystins If those techniques are ready in time for the 2005 field season, they can easily be incorporated into our existing sampling scheme Objective 3: Chemical diversity of microcystins: The individual toxin congeners are determined by HPLC coupled with PDA and MS detection In most cases, identification of the molecular weight and UV absorption pattern is sufficient to identify the microcystin congener In some cases, we must isolate the individual toxin and run either amino acid analysis after hydrolysis (Waters AccQTag) or high resolution NMR (600 MHz) to identify the individual toxins Both capabilities are present at SUNY-ESF Project Relevance: This project represents a significant collaborative venture between NOAA’s MERHAB-LGL regional study and the NOAA-GLERL laboratories Both projects are focused in part on the occurrence of harmful algal blooms in Lake Erie However there are also some major differences and synergies The MERHAB project is specifically focused on monitoring strategies It has a wider focus and includes Lakes Ontario and Champlain in addition to Lake Erie Each lake has a different trophic status and each offers unique challenges MERHAB-LGL is charged with developing and testing new analytical methodology for toxin determination In these regard, we have developed high-throughput methods for toxin analysis and invested three years refining our analytical methodology to accurately measure the concentrations of cyanobacterial toxins in all three lakes (Boyer et al 2004a) We routinely analyze samples submitted by a number of different government agencies and have CDC and EPA approved QA/QC protocols The spatial and seasonal toxin distribution maps proposed here should be directly of interest to several GLERL researchers (G Fahnenstiel, G Leshkevich, and H Vanderploeg) interested in Lake Erie harmful algal blooms Collaboration and other Project Linkages: This project has the potential for a number of collaborative linkages First, the detailed toxin information provided here will be available to all participants in the International Field Year for Lake Erie Since Lake Erie contains both toxic and non-toxic species of cyanobacteria, having the actual toxin concentrations, in addition to any phytoplankton information, is essential to properly interpret any modeling and food web studies As the overall goal of MERHAB-LGL is to develop monitoring and event response strategies for HAB’s on the great lakes, we also have contacts with several water supply providers and health agencies interested in these results Second, since MERHAB-LGL will be supporting the personnel and basic supply costs for toxin analysis, the possibility certainly exists that we can also run samples for toxin analysis collected by other participates in the International Field Year on a no-cost or greatly reduced cost basis (see budget section) Government & Societal Relevance with Implications for Risk Management: Most of our information concerning cyanobacterial toxicity in Lake Erie has focused on the high biomass events that occur in the western basin While these events certainly are important in terms of their impact on drinking water and recreational users of the western basin, the central and eastern basin also serve as drinking water supplies and provide recreational opportunities This study will build on preliminary studies (Ouellette et al, 2005, Boyer et al, in preparation) that show the occurrence of toxic species and cyanobacterial toxins in all three basins of Lake Erie Furthermore, most of our monitoring and toxin detection is in response to a high biomass G.L Boyer page event We have very little information in terms of cyanobacterial toxicity in the early stages of a bloom event For example, recent work on Lake Ontario suggests that toxic August blooms may originate off Toronto in July and be transported by summer lake circulation to the eastern shoreline near Oswego (Boyer et al, 2004a) Similar information is lacking for input into Lake Erie models in that we not have the multiple whole lake time series (Objectives and 2) early in the summer, or the detailed congener/molecular analysis for different bloom events (Objective 3) necessary to track populations This proposal will address those shortcomings A second key point addressed by this proposal is the occurrence of the neurotoxin anatoxin-a Studies in Lake Champlain have shown that the occurrence of anatoxin-a in the water column often has a very different distribution than microcystins First, the anatoxin-a -containing blooms are often low biomass events that would not necessarily trigger reactive sampling Second, the anatoxin-a producing species often occur earlier in the season before the high biomass Microcystis events For example, dog deaths in Lake Neahtawanta (2004) and Lake Champlain (2000, 2001) both occurred in early June (Boyer et al, 2004, unpublished) and were likely due to Anabaena species Microcystis was a minor component in the water column at that time as it usually does not form high biomass blooms until later in the summer Anatoxin-a was found in 12% of the samples we have analyzed from Lake Erie (Yang et al, 2005), but we not at this time have sufficient information to map either its distribution on a whole lake basis, determine the causative organism(s), or evaluate its potential risk to lake users References Brittain, S M., J Wang, L Babcock-Jackson, W W Carmichael, K L Rinehart, and D A Culver (2000) Isolation and characterization of microcystins, cyclic heptapeptide hepatotoxins from a Lake Erie Strain of Microcystis aeruginosa J Great Lakes Res 26:241-249 Boyer, G L., and G D Goddard (1999) High Performance Liquid Chromatography (HPLC) coupled with Post-column electrochemical oxidation (ECOS) for the detection of PSP toxins Natural Toxins 7:353-359 Boyer, G L., J C Makarewicz, M Watzin, and T Mihuc (2004a) Monitoring strategies for harmful algal blooms in the lower great lakes; Lakes Erie, Ontario and Champlain, USA Abstracts, 11th Internat Conference on Harmful Algae Capetown, South Africa, November 15th, 2004 Boyer, G., M C Watzin, A D Shambaugh, M F Satchwell, B R Rosen, and T Mihuc (2004b) The occurrence of cyanobacterial toxins in Lake Champlain In: "Lake Champlain: Partnerships and Research in the New Millennium T Manley, P Manley, T Mihuc, Eds., Kluwer Acad, p 241-257 Carmichael, W W and J An (1999) Using an enzyme linked immunosorbent assay (ELISA) and a protein phosphatase inhibition assay (PPIA) for the detection of microcystins and nodularins Natural Toxins 7:377-385 Harada, K (1996) Chemistry and detection of microcystins In: "Toxic Microcystis" M F Watanabe, K Harida, W W Carmichael, and H Fujiki, Eds., CRC Press, Boca Raton, FL, pp 103-148 James, K J., A Furey, I R Sherlock, M A Stack, M Twohig, F B Caudwell, and O M Skulberg (1998) Sensitive determination of anatoxin-a, homoanatoxin-a and their degradation products by liquid chromatography with fluorimetric detection J Chromatogr A 798:147-157 Li, R., W W Carmichael, S Brittain, G K Eaglesham, G R Shaw, A Mahakhant, N Noparatnaraporn, W Yongmanitchai, K Kaya, and M M Watanabe (2001) Isolation and identification of the cyanotoxin cylindrospermopsin and deoxy-cylindrospermopsin from a Thailand strain of Cylindrospermopsis raciborskii (Cyanobacteria) Toxicon 39:973-980 G.L Boyer page Ouellette, A J A., S M Handy, and S W Wilhelm (2005) Toxic Microcystis is widespread in Lake Erie: PCR detection of toxin genes and molecular characterization of associated cyanobacterial communities Microbial Ecology submitted Yang, X and G.L Boyer (2005) Occurrence of the cyanobacterial neurotoxin, anatoxin-a, in lower Great Lakes Abstracts, International Assoc Great Lake Research Annual Meeting, Ann Arbor MI, May 2005 Project timeline: The regional MERHAB-LGL project continues through 2007 however, it is envisioned that all sample collection associated with this Lake Erie project will occur in 2005 Data analysis should be completed by spring 2006 A brief timeline is given below: MAY APRIL MARCH FEB JAN DEC NOV ? X X X X X X X X X X X X X X Task 3: Identification of Congeners Task 4: Final and interim reports OCT SEPT AUGUS JULY Task 2: Sample analysis JUNE Task 1; Sample Collection MAY 05 Tasks X X X X Budget Request This is a no-cost proposal to NOAA-GLERL as the project is already funded through the NOAA-MERHAB-LGL project MERHAB-LGL will support all the sample analysis and toxin determinations associated with the determination of the spatial and temporal distribution of the toxins in Lake Erie (Objectives and 2), as well as pay for the needed mass spectroscopy time to determine the individual toxin congeners (Objective 3) It will also support travel costs to and from the point of debarkation What is requested is space on board ship for at least two scientists for ~ one week each month (May, June, July, August, Sept, Oct,) In addition, if other investigators participating in the International Field Year for Lake Eire need cyanobacterial toxin analysis for specific samples, the MERHAB-LGL analytical group will attempt to run a reasonable number of those samples either without charge (PPIA, HPLC) or for the cost of supplies only (ELISA; ca $400 per 40 sample plate) Tissue samples represent a special case as standardized extraction techniques often not give an accurate measure of the total toxin content of the tissue We will work with individual investigators to provide high quality results, but NOAA-GLERL should realize that each sample matrix often becomes an individual experiment in itself Depending on the matrix, it simply may not be possible to run those without incurring significant expenses We are also open to discussing any type of at-cost reimbursement if NOAA-GLERL expects a significant number of specialty samples and will work with individual investigators to solve their analytical needs G.L Boyer page Projected Vessel Time Needs Whole lake surveys usually require a large ship deployed for a week at a time We are therefore requesting space for two scientists on a one-week whole lake cruise for the months of May, June, July, August, September and October No specific times are required though it would be best if the cruises are spaced equally apart We would also consider supplemental cruises in August on a 2-week spacing if available or dropping the May cruise if necessary Previous cruises on the Lake Guardian have suggested it is not the best vessel for this work however it is satisfactory We have had excellent results on the RV Limnos and would expect that the RV Laurentian would also be suitable We have some flexibility in that if the ship does not work on a 24 hour schedule, we may be able to get by with fewer (i.e 1) scientists depending upon the ships protocol We also have sufficient equipment that we can double up on two vessels if necessary for simultaneous sampling We have no set stations per se and can easily adapt to others needs In general, the standard monitoring stations (either EPA or Environment Canada) work fine however we need a large number of stations in all three basins to get whole lake coverage Attached is the station map for the RV Limnos in 2004 Coordinates are available from Environment Canada if necessary It would be desirable to have a slightly greater coverage of the western basin There are no special needs other than laboratory space with a sink and the ability to deploy a small (2 lb) submersible pump attached to a 110 volt underwater cable We used this system on the Lake Guardian and Limnos in 2003 without problems A through-hull water system is beneficial in that it allows us to supplement existing stations without stopping the ship, however it is not essential No radioactive material or hazardous waste will be generated or utilized as part of this project Start END G.L Boyer page Curriculum vitae for Gregory L Boyer Faculty of Chemistry, State University of New York College of Environmental Science and Forestry Syracuse, NY 13210 Email: glboyer@esf.edu Telephone: (315) 470-6825 FAX: (315) 470-6855 RESEARCH INTERESTS The chemistry and biochemistry of biologically active natural products from plants and algae including toxins, siderophores, allelopathic agents, and growth regulators Special interests include the biochemistry of iron in forest and aquatic (marine and freshwater) ecosystems, the chemistry / ecology of marine and freshwater harmful algal blooms, brown tides, and rapid detection methods for toxic cyanobacteria and paralytic shellfish poisoning (PSP) toxins EDUCATION: Ph.D., A.B., A.S., University of Wisconsin - Madison, 1980, (Biochemistry) University of California - Berkeley, 1975, (Biochemistry) Reedley College (Reedley, Calif.), 1973, (Chemistry) PROFESSIONAL EXPERIENCE: 1998-present: Professor of Chemistry, State University of New York, College of Environmental Science and Forestry, (SUNY-ESF) Syracuse NY 13210 1991-1998: Associate Professor of Chemistry, SUNY-CESF 1994 Visiting Scientist, Biology Dept., Woods Hole Oceanographic Institute, Woods Hole, MA 02543 1986-1991 Joint Academic Appointment in the Faculty of Environmental Sciences, SUNYESF 1985-1990 Assistant Professor of Chemistry, SUNY-ESF 1983-1984 Research Associate, Dept of Oceanography, Univ of British Columbia, Vancouver, BC, V6T 1W5 1980-1982 Research Associate, Michigan State University - DOE, Plant Research Labs East Lansing, MI, 48824 1975-1980 Research Assistant, Department of Biochemistry, University of Wisconsin, Madison, WI, 53706 HONORS: NRSA predoctoral trainee - Cell & Molecular Biology (1976-79), Life member; Phi Beta Kappa - Alpha (UC-Berkeley Honor Soc.), Life member; Alpha Gamma Sigma (Reedley College Honor Soc.), International Expert for IAEA (International Atomic Energy Agency) on PSP toxins (1999), Participant in EPA’s “Creating a Cyanotoxin Target List for the Unregulated Contaminant Monitoring Rule” taskforce (2001), Participant in NOAA – Sea Grant’s workshop entitled “Developing a National Plan for Remediation of Harmful Algal Blooms”, Steering committee for “National Plan for Marine Biotoxins-2004; Elected as Treasurer and Nominations Chair to the Northeast Algal Society 1999-2005; Recipient; State University of New York Research Foundations 2003 Award for Excellence in the Pursuit on Knowledge G.L Boyer page 10 Selected Publications in the last four years: Giner, J-L., X Li, and G L Boyer (2001) Sterol composition of Aureoumbra legunensis, the Texas brown tide alga Phytochemistry, 57:787-789 Goddard, G D., and G L Boyer (2001) A comparison of HPLC with electrochemical oxidation, HPLC with chemical oxidation, and the mouse bioassay for the analysis of PSP toxins in shellfish In: "Harmful Algal Blooms 2000", G.M Hallegraeff, S.I Blackburn, C.J Bolch, R.J Lewis, eds., p 261-265 Bates, S S., C Leger, M Satchwell, and G L Boyer (2001) The effects of iron on domoic acid production by Pseudo-nitzschia multiseries In: "Harmful Algal Blooms 2000" S.I Blackburn G.M Hallegraeff, C.J Bolch, R.J Lewis, ed., p 320-323 Nichols, D B., M F Satchwell, J E Alexander, N M Martin, M T Baesl, and G L Boyer (2001) Iron nutrition in the brown tide algae, Aureococcus anophagefferens: Characterization of a ferric chelate reductase activity In: "Harmful Algal Blooms 2000", G.M Hallegraeff, S.I Blackburn, C.J Bolch, R.J Lewis, ed., p 340-343 Baker, T R., G J Doucette, C L Powell, G L Boyer, and F G Plumley (2003) Characterization of fluorescent compounds from Pseudomonas stutzeri SF/PS and Pseudomonas /Altermonas PTB-1, bacteria associated with Alexandrium sp and paralytic shellfish poisoning Toxicon 41:339-347 Giner, J.-L., J.A Farldos, G.L Boyer (2003) Unique sterols of the toxic dinoflagellate Gymnodinium breve and a proposed defensive function for unusual marine sterols, J Phycol 39:1-6 Satchwell, M F., and G L Boyer (2003) Comparison of three methods for the detection of microcystin cyanobacterial toxins: Proceedings, 10th International Conference on Harmful Algal Blooms (XHAB), in press Patchett, E.A M.F Satchwell, J Alexander and G.L Boyer (2003) The effects of iron limitation on growth and PSP toxin production in Alexandrium fundyense Proceedings, 10th International Conference on Harmful Algal Blooms (XHAB), in press Boyer, G., M C Watzin, A D Shambaugh, M F Satchwell, B R Rosen, and T Mihuc (2004) The occurrence of cyanobacterial toxins in Lake Champlain In: "Lake Champlain: Partnerships and Research in the New Millennium T Manley, P Manley, T Mihuc, Eds Kluwer, p 241-257 Lehman, P W., S Waller, G Boyer, and K Gehrts (2004) Distribution and toxicity of a new Microcystis aeruginosa bloom in the upper San Francisco Bay region Technical Report for NOAA Coastal Ocean Program Monitoring and Event Response for Harmful Algal Blooms 17p Lehman, P., G Boyer, C Hall, S Waller, and K Gerhts (2004) Distribution and toxicity of a new colonial Microcystis aeruginosa bloom in San Francisco Estuary, California Hydrobiology in press Mihuc, T B., G L Boyer, M F Satchwell, M Pellam, J Jones, J Vasile, A Bouchard, and R Bonham (2005) 2002 Phytoplankton community composition and cyanobacterial toxins in Lake Champlain, U.S.A Verh Internat Verein Limnol 29:xxx-xxx., in press Zou, G., and G L Boyer (2005) Synthesis and Properties of different metal complexes of the siderophore desferriferricrocin Biometals, in press Hotto, A., M Satchwell, and G Boyer (2004) Seasonal production and molecular characterization of microcystins in Oneida Lake, New York, USA Environmental Toxicology in press G.L Boyer page 11 Rinta-Kanto, J M., A J A Ouellette, M R Twiss, G L Boyer, T Bridgeman, and S W Wilhelm (2005) Quantification of toxic Microcystis spp during the 2003 and 2004 blooms in western Lake Erie using quantitative real-time PCR Environ Sci Technol accepted Current and Pending Support Investigator: Gregory L Boyer Home Institution: State University of New York, College of Environmental Science and Forestry Support: √ Current Pending Submission Planned in Near Future Project/Proposal Title: Unrestricted Support for Biochemistry Source of Support: Multiple Sponsors Total Award Amount: $59,750 Total Award Period Covered: 1/1/99 – 12/31/05 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 09 Sumr: Support: √ Current Pending Submission Planned in Near Future Project/Proposal Title: MERHAB 2002: Tier-Based Monitoring for Toxic Cyanobacteria in the Lower Great Lakes Source of Support: NOAA Total Award Amount: $3,622,350 Total Award Period Covered: 9/1/02 – 8/31/07 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: Sumr: 1.5 wk Support: √ Current Pending Submission Planned in Near Future Project/Proposal Title: New York State Disinfection By-Product/Algal Toxin Project Source of Support: NYS-DEC Total Award Amount: $23,000 Total Award Period Covered: 6/15/04 – 5/31/05 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 09 Sumr: Support: √ Current Pending Submission Planned in Near Future Project/Proposal Title: Central New York Biotechnology Research Center Source of Support: US Department of Health and Human Services Total Award Amount: $713,087 Total Award Period Covered: 9/1/03 – 12/31/08 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 09 Sumr: Support: √ Current Pending Submission Planned in Near Future Project/Proposal Title: Source Sentinel : Protection of Municipal Water Supplies Source of Support: NYS Office of Science, Technology and Academic Research Total Award Amount: $300,000 Total Award Period Covered: 4/1/02 – 12/31/05 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 45 Sumr: Support: √ Current Pending Submission Planned in Near Future Project/Proposal Title: NSF Graduate Research Fellowship for Karen Howard Source of Support: National Science Foundation Total Award Amount: $40,500 Total Award Period Covered: 6/1/04 – 5/31/05 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 09 Sumr: NSF Form 1239 (10/98) USE ADDITIONAL SHEETS G.L Boyer page 12 AS NECESSARY G.L Boyer page 13 Current and Pending Support Investigator: Gregory L Boyer Continued Support: Current √ Pending Submission Planned in Near Future Project/Proposal Title: Spatial and temporal Distribution of Cyanobacterial Toxins in Lake Erie (This proposal) Source of Support: NOAA-GLERL Total Award Amount: $0 no cost Total Award Period Covered: 5/1/2005 – 4/30/2006 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 0.09 Sumr: Support: Current √ Pending Submission Planned in Near Future Project/Proposal Title: Technical Evaluation of a Near-real time monitoring platform for drinking water contaminants Source of Support: New York Indoor Environmental Quality Center Total Award Amount: $25,000 Total Award Period Covered: April 1, 2005 – 12/31/2005 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 0.09 Sumr: mo Support: Current √ Pending Submission Planned in Near Future Project/Proposal Title: This Proposal: A Remote-Sensing Based Early Warning System for Algal Blooms in the Great Lakes Source of Support: University of Buffalo/NASA Total Award Amount: $182,226 Total Award Period Covered: 10/1/05 – 9/30/08 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 36 Sumr: mo Support: Current √ Pending Submission Planned in Near Future Project/Proposal Title: The Health and Ecosystem impacts of a newly developed cyanobacterial bloom of Microcystis in the Sacramento-San Joaquin Delta Source of Support: California Department of Environmental Conservation Total Award Amount: $124,612 Total Award Period Covered: 7/1/2006 – 7/1/2009 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 0.36 Sumr: Support: Current Pending √ Submission Planned in Near Future Project/Proposal Title: Deployment of an automated system for the detection of toxic cyanobacteria Source of Support: New York Sea Grant Total Award Amount: $74.028 Total Award Period Covered: 2/1/2006 – 1/31/2007 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 0.45 Sumr: Support: Current Pending √ Submission Planned in Near Future Project/Proposal Title: Historical Distribution of Microcystis and its Toxins in Lake Erie Sediments Source of Support: NOAA-GLERL Total Award Amount: $19,500 Total Award Period Covered: 5/1/2005 – 4/30/2006 Location of Project: SUNY College of Environmental Science and Forestry, Syracuse, NY Person-Months Per Year Committed to the Project Cal: Acad: 0.45 Sumr: NSF Form 1239 (10/98) USE ADDITIONAL SHEETS AS NECESSARY ... support all the sample analysis and toxin determinations associated with the determination of the spatial and temporal distribution of the toxins in Lake Erie (Objectives and 2), as well as pay for... cyanobacterial toxins in Lake Erie extends beyond the occurrence of microcystins in the western basin Our working hypothesis is that the occurrence of microcystins in the eastern and central basins will... Occurrence and Distribution of Cyanobacterial Toxins in Lake Erie Cruise and Date # samples % containing Highest Comments toxin measured value * Brittain et al 44 MC’s ~10% 3.4 µg L-1 MC Western basin