Harmful Algal Blooms in Lake Ontario

Home , Microcystis, Microcystis aeruginosa

Cyanobacteria and their Toxins in NY and Great Lakes Waters

Greg Boyer ([email protected]) State Univ. New York- College of Environmental Science and Forestry (ESF) in Syracuse NY Faculty of Chemistry Director, Great Lakes Research Consortium

M ERHAB-LGL Harmful Algal Bloom M oni tor i ng and Ev ent Response i n the Low er Gr eat Lak es Sodus Bay September 2010 Lake Erie 2009

Lake Champlain 2008 Things to cover

• Introduction to the toxins

• Large Lakes – Lake Champlain – Lake Ontario – Lake Erie • Inland lakes of New York

• Take home message: not that different Microcystins • Microcystis aeruginosa • non-N fixer. • Very common – Also produced by a number of other species.

D-Glu (iso) Methyl dehydroAla (Mdha) CH3 CH N 2 CO2H • Peptide hepato-Toxin: 90+ structural variants + R O NH NH O R Adda O CH3 100-200 other bioactives D-Ala H3C S (anabaenapeptins, etc.) OCH3 O NH O S S 1 NH CH CH3 3 O S R NH Leu CH3 CH3 S CH3 • Called “fast death factor” NH O CO2H 2 – LD-50: 25-60 µg kg-1 MeAsp (iso) Arginine Potent carcinogen NH NH2 NH • WHO guideline value is 1 µg/L for drinking water Never trust a name! Microcystin-producing strains include:

• Microcystis • Planktothrix aeruginosa agardhii • M. veridis • P. mougeotii • M. botrys • Nostoc spumigena • N. species • Oscillatoria limosa • Anabaenopsis • Anabaena flos-aquae millerii • A. lemmermannii • Haphalosiphon • A. circinalis hibermicus i.e. Biology is a mess! no no no

yes yes

yes

no yes yes no Difficult to use taxonomy to predict toxicity

So how do you tell who is making the toxin?

DNA based probes!

CYA

mcyB mcyD Mic Tillett et al, 2000 ImageOuellette courtesy and Wilhelm of S. Wilhelm; 2003, 2005 Microcystin Biosynthetic Gene Cluster

Microcystis aeruginosa PCC 7806

J I H GK GP F EP EK D A1 A2 B1 B2 C

Planktothrix agardhii CYA126/8

T D EK EP GP GK H A1 A2 B1 B2 C J

Anabaena sp. 90

H I F EP EK J D GK GP A1 A2 B1 B2 C LE03-WLE1-A07 McyA sequences LE03-WLE1-A12 LE04-882-F02 LE03-WLE1-A05 Microcystis sp. TuM7C Microcystis aeruginosa K-139 Microcystis aeruginosa UV027 Microcystis aeruginosa PCC7806 LE03-WLE1-C04 Microcystis aeruginosa PCC7941 52 LE03-WLE1-D01 Microcystis sp. IZANCYA5 57 LE04-974-C02 LE04-974-D02 64 LE04-974-C11 53 LE03-WLE1-A01

100 LE04-974-C04 Microcystis aeruginosa LE-3 2003 Microcystis aeruginosa NIES-89 LE04-974-C08 2004 LE03-WLE1-D02 LE04-974-B12 LE04-974-B11 50 LE04-974-B09 LE04-974-E05 LE04-974-C05 Nostoc sp. IO-102-I Nostoc sp. 152 2 different Anabaena circinalis 90 98 Anabaena flos-aquae N IVA-CYA83 populations LE04-1163-H05 producing the same 98 LE04-1163-A04 70 LE03-1163-D04 toxin!! Planktothrix agardhii CYA126/8 LE04-1163-H07 53 LE03-1163-E04 Streptomyces verticillus ATCC15003

0.1 Rinta-Kanto and Wilhelm, 2006, AEM 72:5083 Changes in total and toxic Microcystis in Lake Erie

2005 109 July August 108 Cyanob.

107 -1 106 Total M.

5 Cells L Cells 10

104 Toxic M.

103

102 357 478 558 882 971 972 973 974 968 969 e58 e59 e60 344 357 358 469 478 558 580 835 865 881 882 885 971 972 973 974 968* 969* 973* 1163 1163 e91M 478A 478A 1163* N.a.N. Station

Toxigenic Microcystis can account for >95% of the total Microcystis Toxigenic Microcystis can account for <10% of the total Microcystis

These numbers can change with time. Rinta-Kanto et al, Harmful Algae, 2009 Cyanobacteria blooms in the Great Lakes

M ERHAB-LGL Harmful Algal Bloom M oni tor i ng and Ev ent Response i n the Low er Gr eat Lak es

From Watson et al., 2008 NY TotalsNY Ontario L. Erie L. Champlain L. Finger Lakes LakesLocal Occurrence of Microcystins as measured by PPIA (2000 – 2004)

# Analyzed: 2513 736 308 590 138 741

> 0.01ug/L 1223 155 117 296 113 542 (53%) (28%) (40%) (51%) (82%) (73%)

> 0.1 ug/L 829 61 84 190 23 471 (36%) (14%) (29%) (33%) (17%) (64%) > 1 ug/L 326 4 11 71 1 239 (14%) (1%) (4%) (12%) (1%) (32%)

Boyer, Lake Res. Managem, 2007 Anatoxin-a

. Potent Neurotoxin. • (very fast Death Factor) . LD-50: 200 µg kg-1 O . Causative organisms include: NH2Cl • Anabaena species (many) • Oscillatoria sp. • Aphanizomenon sp. • Planktothrix sp. • Responsible for the Lake Champlain animal fatalities in 2000 and 2001. CH3 N CH3 Anatoxin-a(S) HN Organophosphate N O Saxitoxin Family Neurotoxin. NH + O 2 - P Potent neurotoxins cholinesterase O OCH3 Responsible for inhibitor Paralytic Shellfish LD-50: 20 µg kg-1 Poisoning (PSP) -1 LD-50: 10 µg kg Cylindrospermopsin NH2 O LD-50: 300 µg kg-1 H O NH HN Causative species: OH NH + - 2 C. raciborskii O3SO O + NH NH2 N OH Aph. ovalisporum N NH HN NH CH3 OH Umezakia natans NH+ O Very common in Florida

To date, these other toxins are very rare in our region…….. NY TotalsNY Ontario L. ErieL. Champlain L. Finger Lakes LakesLocal Data Mining Other Toxins

Thresholds and samples number

ATX (>0.1 ug/L) 29 2 2 12 2 11 (n>3,000) (1%) (<1%) (1%) (2%) (1%) (2%) ATX (>0.01 ug/L) 75 14 14 24 2 21 (n>3,000) (3%) (5%) (5%) (4%) (1%) (3%) CYL (>0.01ug/L) 8 ? 1 ? 2 ? 0 - 5 ? (n>2,500) (<1%) (<1%) (0%) (0%) (2%) PSP (>0.01ug/L) 2 0 1 0 0 1 (n>2,500) (0%) (0%) (0%) (0%) (0%) (1%)

Modified from Boyer, 2007 What about ATX Distribution?

Distribution of Anatoxin-a in Lake Ontario (n= 940) • Widespread • Ephemeral • Not correlated with Microcystis - different genus - different ecology. Who is the producer ? Anatoxin-a tended to be localized in the embayments.

Sandusky Bay v.s. Sandusky basin Toledo ------>Lake

Maumee River 2004 Stability of Anatoxin-a influenced by pH and light intensity

12 hr experiments Survey of 62 water bodies throughout NYS:

• Presence of Cyanobacteria (77%) • Presence of toxic species (Microcystis 40%) • Presence of toxin genes (50%) • Presence of toxins (50% with 5% >1ug/L)

– Toxic – Non-toxic – Potentially toxic (Toxigenic) M ERHAB-LGL Harmful Algal Bloom M oni tor i ng and Ev ent Response i n the Low er Gr eat Lak es • Correlate this with easily- to-measure parameters. Hotto et al., 2007 Most blooms were associated with shallower waters

20 1.0 15

0.8 10 gene Chlorophyll-a (ug/L) Chlorophyll-a

0.6 5

0 0.4 0 10 20 30 40 50 60 70 80 90 Mean Lake Depth (m) 0.2 Probability of the mcyB the of Probability

0.0 0 50 100 150 200 Temp >21° Max Lake Depth (m) Good correlation between nutrients and algal biomass

20 There was a weak correlation 15 between the 10 presence of

Chlorophyll-a (ug/L) Chlorophyll-a 5 toxins and nutrients 0 0 50 100 150 200 250 Total Phosphorus (ug/L) Toxin ==X growth Increased biomass increased the chance of a toxigenic bloom

1.0

0.8 Oligrotrophic waters <5% gene

0.6 Mesotrophic waters ~20-40%

0.4 Hypereutrophic waters >90%

0.2 Source water protection works! Probability of the mcyA the of Probability

0.0 0 10 20 30 40 50 60 Chlorophyll-a Concentration (ug/L) Lake Neatahwanta

Lake Neatahwanta 2004-2007 50 Hepatotoxins

40 They are gone!

30 Why did it go away?

20 Will it come back? algal toxins in ug/L in toxins algal

0 Mar- Jun- Sep- Dec- Mar- Jun- Sep- Dec- Mar- Jun- Sep- Dec- Mar- Jun- Sep- Dec- Mar- 04 04 04 04 05 05 05 05 06 06 06 06 07 07 07 07 08 Date Lake Neatahwanta

Lake Neatahwanta 2004-2007 50 Hepatotoxins August 2010: 3 ug/L

40 May 2010 600 ug/L

20 algal toxins in ug/L in toxins algal

0 Mar- Jun- Sep- Dec- Mar- Jun- Sep- Dec- Mar- Jun- Sep- Dec- Mar- Jun- Sep- Dec- Mar- 04 04 04 04 05 05 05 05 06 06 06 06 07 07 07 07 08 Date Summary • Cyanobacteria produce a number of toxins but not all species are toxic. – Toxic species, potentially toxic species and non-toxic species • Hepatotoxic microcystins are probably the toxin of most concern for human health in New York State. • These toxins can be produced by a number of different species making visual monitoring difficult. • We do not understand the spatial, temporal and environmental factors affecting cyanobacterial toxin production. • Source water protection remains our best tool for controlling the incidence of toxic cyanobacterial blooms within New York State. People who did the work: SUNY-ESF, Syracuse Univ. of Tennessee • Mike Satchwell (Forestry) • Steven Wilhelm (Molecular biol) • Margaret Pavlac (Environ. Chem.) • Johanna Rinta-Kanto • Jeremy Sullivan (Biochemistry) • Jenn DeBruyn • George Westby (GLOS liason) • Katherine Perry (Environ. Chem.) Lake Champlain. • Tim Mihuc (LCRI) Gone by not forgotten • Mary Watzin (UVM) • Amber Hotto (Biochemistry) SUNY-Brockport • Xingye Yang (Biochemistry) • Pauline Stevens (Remote sensing) • Joe Makarewicz • Juliette Smith (Fish biology) SUNY-Buffalo • Karen Howard (Analytical Chem.) • Joe Atkinson • Elizabeth Konopko (Environ. Chem.) • Steve Ragonese (technical support) SUNY Stony Brook • Guozhang Zou (biochemistry) • Chris Gobler • Sean Thomas (Environmental Science) • Tim Davis Environment Canada • A whole host of undergraduates! • Susan Watson • Captain & Crew of CCGS Limnos