Cyanobacteria and their Toxins Real Concern or Just Green Water?

Nicolas Clercin

Research Scientist, IUPUI Center for Earth and Environmental Science

nclercin@iupui.edu

What is an algal bloom?

NASA’s Aqua/MODIS Satellite – Oct, 9th 2011

Grand Lake St Marys, OH July 2010 (EPA)

Geist Reservoir, IN Summer 2011 (Environmental Law & Policy Center)

• Anoxia (Oxygen Depletion) – Fish kills – Changes in freshwater community structure

• Degradation of recreational resources – Socio-economic impacts – Property value impacts – Recreational use limitation

• Taste and odor in drinking water – MIB (musty) and Geosmin (earthy smell)

• Algal Toxins – Fish and other aquatic life – Livestock – Pets (reports of dog deaths in Indiana, 2009) – Humans

Impacts Associated with Algal Blooms

Fox River, IL 2005

Palestine Lake, IN

University Lake, IN

• Kingdom: Prokaryota – Cyanobacteria or “blue-green algae”

• Prokaryotic algae lacking typical membrane-bound nuclei and plastids • Capable of photosynthesis (Photobacteria) • Gram- bacteria like E. coli* (gastroenteritis), Salmonella* (typhoid fever),

Shigella* (dysentery), Pseudomonas (nosocomial infections), Neisseria (meningitis), Bordetella pertussis, Vibrio cholerae…

• Kingdom: Eukaryota

– Eukaryotic algae with typical nucleus and pigments localized within plastids.

• Chlorophytes (green algae) • Bacillariophytes (diatoms) • Cryptophytes (cryptomonads) • Pyrrhophytes (dinoflagellates) • Chrysophytes (golden algae) • Euglenophytes (euglenoids)

Algae classification

*Microbial Contaminants in EPA Contaminant Candidate List 3 (CCL3) http://www.epa.gov/safewater/ccl/index.html

Cyanobacteria • Chroococcales

– Solidary or colonial coccoid “blue-greens”, never form true filaments • Include: Aphanocapsa, Aphanothece, Chroococcus, Coelosphaerium, Gloeocapsa, Gloeothece,

Gomphosphaeria, Hyella, Merismopedia, Microcystis, Pleurocapsa, Synechococcus, Woronichinia

• Oscillatoriales – Filamentous colonies, no heterocyst- and akinete-formation

• Include: Homeothrix, Limnothrix, Lyngbya, Oscillatoria, Phormidium, Planktothrix, Plectonema, Pseudanabaena, Schizothrix, Spirulina, Trichodesmium

• Nostocales – Filaments forming heterocysts and akinetes, but never true branches

• Include: Anabaena, Anabaenopsis, Aphanizomenon, Calothrix, Cylindrospermun, Cylindrospermopsis, Gloeotrichia, Nostoc, Rivularia, Scytonema

• Stigonematales – Filaments forming heterocysts and true branches

• Include: Brachytrichia, Desmosiphon, Fischerella, Hapalosiphon, Stigonema

Aphanocapsa Merismopedia Lyngbya Spirulina Anabaena Gloeotrichia Fischerella Stigonema

Chroococcales Oscillatoriales Nostocales Stigonematales

• Hepatotoxins: disrupt proteins that keep the liver functioning, may act slowly (days or weeks later) Microcystins (>80 structural variants)

• LD50: 25-60 μg kg-1

• Potent carcinogen (liver, kidneys) Microcystis aeruginosa, M. viridis Anabaena flos-aquae, A. circinalis Planktothrix agardhii, P. mougeotii Oscillatoria limosa Nostoc sp. Anabaenopsis millerii Aphanocapsa

Nodularins (9)

• LD50: 60 μg kg-1

Nodularia spumigena

Cylindrospermopsins (3)

• LD50: 300 μg kg-1 Aphanizomenon ovalisporum Cylindrospermopsis raciborskii Umezakia natans

Toxicity - Cyanotoxins

Microcystin-LR

Cylindrospermopsin

• Neurotoxins: cause rapid paralysis of skeletal and respiratory muscles Anatoxin-a (5)

LD50: 200-250 μg kg-1

Anabaena flos-aquae, A. planctonica Anabaena spiroides, A. circinalis Aphanizomenon sp. Cylindrospermum sp. Microcystis sp. Oscillatoria sp.

Anatoxin-a(S) (1)

LD50: 20 μg kg-1

Anabaena flos-aquae, A. lemmermannii

Saxitoxins (20) LD50: 10 μg kg-1 Potent neurotoxin; Paralytic Shellfish Poisoning (PSP)

Anabaena flos-aquae, A. lemmermannii Aphanizomenon flos-aquae Cylindrospermopsis raciborskii Lyngbya wollei Planktothrix agardhii

β-N-methylamino-L-alanine (BMAA), neurodegenerative agents, causative of amyotrophic

lateral sclerosis (Lou Gehrig’s Disease) and parkinsonism-dementia complex (ALS/PDC)

Toxicity - Cyanotoxins

BMAA

Saxitoxin

Anatoxin-a

Toxicity - Cyanotoxins • Dermatotoxins: produce rashes and other skin reactions (hours or days). Potent

irritants, vesicants and carcinogens. Lyngbyatoxin-a (1)

• Potent irritant and vesicant, aka seaweed dermatitis Moorea producta (ex- Lyngbya majuscula)

Aplysiatoxins (2) • Potent tumor promoters

Lyngbya majuscula

• Endotoxins: inflame gastrointestinal epithelium. They are pyrogenic (fever) and

induce immune response. At high enough concentrations in blood, endotoxins are lethal (septic shock). Lipopolysaccharides (LPS)

Lyngbyatoxin-a

Toxins and Taste & Odor Compounds Produced by Cyanobacteria

Species

Dermatotoxin, Endotoxin

Hepatotoxin (Liver)

Neurotoxin (Nervous System)

Taste/Odor Compound

Aphanacapsa spp. LPS microcystins

Microcystis spp. LPS microcystins, nodularin anatoxins

Snowella spp. LPS microcystins

Synechococcus spp. LPS microcystins MIB, Geosmin

Woronichinia spp. LPS microcystins

Lyngbya spp. LPS, lyngbyatoxins saxitoxins MIB, Geosmin

Oscillatoria spp. LPS, aplysiatoxins microcystins anatoxins, saxitoxins MIB, Geosmin

Planktothrix agardhii LPS, aplysiatoxins microcystins saxitoxins MIB, Geosmin

Pseudoanabaena spp. LPS MIB, Geosmin

Anabaena spp. LPS microcystins

cylindrospermopsin anatoxins saxitoxins

MIB, Geosmin

Anabaenopsis elenkinii LPS microcystins

Aphanizomenon spp. LPS cylindrospermopsin

microcystins anatoxins saxitoxins

Geosmin

Cylindrospermopsis raciborskii LPS cylindrospermopsin saxitoxins

Nodularia spp. LPS microcystins ,nodularin

LPS = Lipopolysaccharide Modified from: Graham et al., 2008; Lawton and Edwards, 2008; NOAA, 2007; Graham, 2007; Wiedner et al., 2006

• Currently, there are NO US Federal guidelines, Water Quality Criteria and Standards or Regulations concerning harmful algal blooms.

• Cyanobacterial toxins are on EPA CCL3; EPA is reviewing standards and risks. • World Health Organization* established guidelines for drinking waters (1998)

and recreational use of water bodies (1999), based on: – Cell concentrations and/or biovolume; – Chlorophyll-a (equivalent to cell levels); – Microcystin.

• Several states have statewide monitoring programs and well-defined public health reporting and response systems – Indiana is following this path.

Status of Harmful Algal Bloom Guidelines and Standards

*WHO (1998) Guidelines for Drinking–Water Quality. Second Edition, Addendum to Volume 2, Health Criteria and Other Supporting Information. World Health Organization, Geneva. *WHO (1999) Toxic Cyanobacteria in Water: A guide to their public health consequences, monitoring and management. I. Chorus and J. Bartram (eds.), E & FN Spon, New York, 416

WHO Guidance Values for Probability of Acute Health Risks During Recreational Exposure to Cyanobacteria and Microcystin-LR

Hazard Status Cyanobacteria Microcystin – LR Recommended Action

Low

< 20,000 cells. mL-1 or < 10 μg. L-1 chlorophyll-a or < 2.5 mm3 L-1 cyanobacterial biomass

<10 µg/L Post Advisory Signs and Continue Monitoring

Moderate

20,000 - 100,000 cells. mL-1 or 10 - 50 μg. L-1 chlorophyll-a or 2.5 - 12.5 mm3 L-1 cyanobacterial biomass

10 - 20 µg/L Post Advisory Signs and Restrict Swimming

High

> 100,000 cells. mL-1 or > 50 μg. L-1 chlorophyll-a or > 12.5 mm3 L-1 cyanobacterial biomass

20 - 2,000 µg/L Post Advisory Signs, Prohibit

Swimming and Other Water-contact Activities

US States with Reports of Acute Cyanobacterial Poisonings*

*Graham, J. L. , K.A. Loftin, and N. Kamman. Monitoring Recreational Freshwaters. Lakeline. Summer 2009. pp 18-24

Cyanobacterial Poisonings and Health Advisories Cyanobacterial Poisonings Health Advisories

Central Indiana Reservoirs 2008-2011

• Cell Density: – Moderate risk from

April to June – High risk from July to

November

• Biovolume: – Always at high risk!

• Microcystin: – Toxic from April to

January – Close to Indiana

recommendation from July to October

Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Den

sity

(cel

ls/m

L)

101

102

103

104

105

106

107

Low Risk

Moderate Risk

High Risk

Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Bio

volu

me

(mm

3 /L)

100

101

102

103

104

105

Low Risk

Moderate Risk

High Risk

Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Mic

rocy

stin

( g/

L)

10-2

10-1

100

101

102

Recreational Water

Drinking Water

State of Indiana

Detection Limit

2008-2011 Occurrences of Microcystin as Measured by Elisa-Adda in Central Indiana Reservoirs

Geist

0.0

2.0

4.0

6.0

8.0

Morse

Mic

rocy

stin

( g/

L)

0.0

1.0

2.0

3.0

Eagle Creek

Jan-08 Jul-08 Jan-09 Jul-09 Jan-10 Jul-10 Jan-11 Jul-11 Jan-12 0.0

1.0

2.0

3.0 Reservoir damUpper reservoir

# Occurrences (% Occurrences)

Central Indiana Eagle Creek Morse Geist

# Analyzed 715 299 202 214

> 0.15 μg/L 365 (51.0%)

97 (34.2%)

116 (57.0%)

152 (71.0%)

> 1 μg/L 82 (11.5%)

5 (1.7%)

13 (6.4%)

64 (29.9%)

> 3 μg/L 16 (2.2%)

0 (0%)

0 (0%)

16 (7.5%)

> 6 μg/L 1 (0.1%)

0 (0%)

0 (0%)

1 (0.5%)

• State of Indiana worked with CEES-IUPUI in 2008 to report results of CEES/Veolia Water monitoring of 3 central Indiana reservoirs

• State of Indiana utilizes 100,000 cells/mL and/or 6 µg/L (ppb) microcystin for posting advisories and press releases – IDEM Website Postings: www.algae.in.gov – ISDH Press Releases

• CEES research programs have continued to provide information to State since 2009 for posting on state website

• CEES now provides information for 3 central Indiana Reservoirs and Patoka Lake (Southern Indiana) since 2010

• IDEM collaborated with CEES-IUPUI in 2010/11 and developed its own statewide monitoring program

Indiana Monitoring and Response to Harmful Algal Blooms

Thank you

NASA’s Aqua/MODIS Satellite – Lake Erie Jan, 15th 2012