7/18/2016

Palmer Prints: Algae in Water Supplies Palmer Algal Posters to Cyanotoxins; • 1959: Algae in Water Supplies changes in our knowledge of • “An Illustrated Manual of the Identification, Significance and Control of Algae in Water cyanobacteria (bluegreens) Supplies” • C. Mervin Palmer, artist Harold J. Walter NC AWWA‐WEA • U.S. Dept. of Health, Education and Welfare Drinking Water Rules & Regulations Seminar 2016 • 1965: Prints put into Standard Methods Mark Vander Borgh, Linda Ehrlich and Astrid Schnetzer • “Standard Methods for the Examination of Water and Wastewater” 12th Ed.

Algae Growing on Reservoir Walls

Palmer Prints: Algae in Polluted Water What are Bluegreens? • Bacteria or Algae? • 1970: USEPA created • Eubacteria • Prokaryotic • 1974: Clean Water Act • no membrane bound organelle • Photosynthetic • 1977: Algae and Water Pollution: • opposed to chemotrophic (i.e., iron bacteria) • “An Illustrated Manual on the Identification, • Various Photosynthetic Pigments Significance and Control of Algae in Water • Chlorophyll‐a (green), phycocyanin (blue) or Supplies and in Polluted Water” phycoerythrin (red) • Two prints added, artist: Sharon Adams • Smallest form of algae • From < 1um (picoplankton) to > 20um • filaments > 100 um long • Colonies, filaments and clumps visible to naked eye

Polluted Water Algae Plankton and other Surface Water Algae

What is the Official Name? Why are they Important?

• Bacteria The Good • Myxophyceae • Primary producers • Oxygen, carbon & carbohydrates • Cyanoprokaryotes • Nitrogen fixing • Cyanophytes • heterocysts/heterocytes • Blue‐greens • Symbiotic forms • Bluegreens • Lichen & Azolla (aquatic fern) •Cyanobacteria Azolla

Filter Clogging Algae Trichormus azollae Clean Water Algae

1 7/18/2016

Why are they important? Importance of Bluegreens in North Carolina

The Bad • Common in North Carolina • Taste and odors • Found in water supplies • Geosmin and Methylisoborneol (MIB) • Many form blooms • Bloom formers • Sub‐surface/stratified • Some are basis for “impairments” • Surface films (AKA scums) • Chlorophyll‐a (>40 ug/L) • requires gas vacuoles (aerotropes) • pH (< 6 or > 9) • Some produce toxins • Some may form cyanotoxins • Toxigenic strains • Harmful Algal Blooms (HAB)

Taste and Odor Algae

Basic Bluegreen Taxonomy

• Prokaryotic • (cytoplasm fills cell) • Color: INTRODUCTION TO • Blue, green, bluegreen, red & gray‐violet • Morphology BLUEGREEN TAXONOMY • Unicellular/colonial; coccoid/filamentous (updating nomenclature) • Heterocytes (N‐fixing cells)

Bluegreen Taxonomy: Lumper vs Splitters Basic Bluegreen Groups (Orders)

• Simplest Revision • Chrooccocales • Drouet & Daily, 1956 “revision of the coccoid and nostocales myxophyceae” • round or oval • Most Thorough • Synechococcales • Anagnostidis & Komarek, 1998 “Suswasserflora von mitteleuropa” • round, oval or filamentous (freshwater flora of central Europe) • Komarek, 2013. Cyanoprokaryota. Heterocytous genera. “Suswasserflora von • Oscillatoriales & Spirulinales mitteleuropa” • filaments without heterocytes • Cyanobacteria nomenclature website http://www.cyanodb.cz/ • Nostocales • Current Primary Reference: • filaments with heterocytes • Wehr, Sheath & Kociolek, 2015 “Freshwater Algae of North America” 2ed

2 7/18/2016

Order: Chroococcales Order: Synechococcales

• Family: Microcystacaeae: • Family: Merismopediaceae • Microcystis (Anacystis & Polycystis) • Merismopedia (Agmenellum) • Family: Aphanothecaceae • Limnococcus = (Chroococcus) • Aphanothece • Aphanocapsa • Family: Romeriaceae Microcystis • Romeria

Limnococcus Romeria Aphanothece Aphanocapsa

Order: Synechococcales Order: Synechococcales (filamentous)

• Family: Coelosphaeriacae All previously called Oscillatoria • Coelosphaerium • Family: Pseudanabaenaceae • Woronichinia (Gomphosphaeria) • Pseudanabaena • Snowella • Limnothrix • Family: Leptolyngbyaceae • Leptolyngbya Limnothrix • Planktolyngbya (nordicmicroalgae.org)

Coelosphaerium Woronichinia (Meredith College) Planktolyngbya (www.algalweb.net) Pseudanabaena Snowella (Robin Mathews)

Order: Spirulinales Order: Oscillatoriales Benthic, attached and/or mat‐forming • Family: Microcoleaceae • Planktothrix (Oscillatoria) • Family: Oscillatoriaceae • Oscillatoria • Lyngbya

Planktothrix

Lyngbya wollei = Plectonema wollei = Microseira? Oscilllatoria

3 7/18/2016

Order: Nostocales Order: Nostocales

• Presence of special cells • Family: Aphanizomenonaceae

• Heterocytes (N2 fixation) • Anabaenopsis ( UNC Inst. Marine Sciences) • Akinetes (protective, reproductive • Cylindrospermopsis (Anabaenopsis) spore) • Raphidiopsis (?)

A

H

(missouristate.edu) Aphanizomenon Anabaenopsis (cfb.unh.edu) Dolichospermum Cylindrospermopsis (Meredith College) Raphidiopsis

Order: Nostocales Order: Nostocales

• Family: Nostocaceae • Family: Aphanizomenonaceae Aphanizomenon raft • Anabaena • Aphanizomenon • Most species reclassified to genetically supported genera • Cuspidothrix (Aphanizomenon) • Genus still exists; mostly benthic • Dolichospermum (Anabaena) • Sensu stricto: cylindrical akinetes, obligately without aerotopes

(Meredith College)

Dolichospermum Cuspidothrix (B. Rosen)

Cyanotoxins: What are they?

• EPA Chemical Contaminants List: • Other Toxins • Microcystin • Saxitoxin • Cylindrospermopsin • Nodularin INTRODUCTION TO • Anatoxin • BMAA • EPA Health Advisories: CYANOTOXINS • Microcystin • 70‐80 Varients • Cylindrospermopsin • 3 Varients

Microcystin (enzolifesciences.com)

4 7/18/2016

Cyanotoxins: Who makes them? Cyanotoxins: What do they effect?

• Microcystins • Microcystin & Cylindrospermopsin: • Anabaenopsis, Aphanizomenon, Dolichospermum, • Hepatatoxin, Cytotoxins, Endotoxin & Dermatoxin Limnothrix, Microcystis, Oscillatoria, Planktothrix, • Liver, kidney, stomach & skin • Kidney and liver damage Woronichina & Snowella • Gastroenteritis, gastrointestinal upsets and skin irritation • Cylindrospermopsin • Anatoxin • Aphanizomenon, Cylindrospermopsis, Lyngbya, • Neurotoxin Raphidiopsis • Nervous system Cylindrospermopsin • tingling, numbness, drowsiness, respiratory paralysis • Anatoxins (www.epa.gov/cyano_habs_symposium/monograph/Ch03.pdf) Anatoxin • Aphanizomenon, Cylindrospermopsis, Cuspidothrix, (http://www.epa.gov/cyano_habs_symposium/monograph/Ch03.pdf) Oscillatoria, Pseudanabaena

Cyanotoxins: Testing Cyanotoxins: Testing • Types of samples: • Particulate (cell bound) • Bioassays (class specific) • Dissolved (exocellular) • use of live animal, tissue or uses biological • Total toxin (particulate + dissolved) activity of a substance • enzyme‐linked immunosorbent assay (ELISA) • mouse assays • Toxin Testing Methods • phosphate inhibition assay (PPIA) • Chromatography (compound specific) (clipartpanda.com) • separation of a mixture by passing it in solution or as a vapor through a medium in which components move at different rates • Can separate toxin congeners/varients ELISA test (hadlemosdeciencia.wordpress.com)

Solid Phase Adsorption Toxin Tracking ‐ SPATT

• A simple and sensitive in situ (monitoring) method • involves the passive adsorption of biotoxins onto porous synthetic resin filled sachets (SPATT bags) and their subsequent extraction and analysis. MacKenzie et al. (2004) Toxicon EPA PROS • Time‐integrative signal HEALTH ADVISORIES • Low detection limit • Fresh to marine application • Multiple toxin detection • Easy to deploy and recover

CONS • Semi‐quantitative • No link to regulatory limit

5 7/18/2016

EPA Health Advisories EPA Health Advisories

• Why Issue a Health Advisory • How are Health Advisories Created? • The occurrence of cyanotoxins within HABs • Review of current literature with a focus on: have increased in distribution and frequency • Occurrence • There are currently not enough health effects • Environmental fate data to support a regulation • Mechanism of toxicity • Toxicokinetics • Acute, short term, subchronic and chronic toxicity • Health effects • Exposure • Analysis and treatment for drinking water

EPA Health Advisories EPA Health Advisories

Cyanotoxin Unsafe Threshold Comment Drinking Water Health Advisory Documents Fact Sheet: 2015 Drinking Water Health Advisories for Two Cyanobacterial Toxins (PDF) Microcystins > 0.3µg/L (ppb) All ages Drinking Water Health Advisory for the Cyanobacterial Toxin Cylindrospermopsin (PDF) Cylindrospermopsin > 0.7 µg/L (ppb) Children < 6 years old Drinking Water Health Advisory for the Cyanobacterial Microcystins Toxins (PDF) Cylindrospermopsin > 1.6 µg/L (ppb) Ages ≥ 6 years old Where can I find more information? Information on the HAs for microcystins and cylindrospermopsin and the health effects support documents for these and anatoxin‐a in drinking water DRINKING WATER ONLY! EPA’s Health Advisory webpage: http://water.epa.gov/drink/standards/hascience.cfm These advisories are based on a 10 day exposure Information on additional strategies Public Water Systems and others could consider in managing cyanotoxins EPA’s CyanoHABs website: http://www2.epa.gov/nutrient‐policy‐data/guidelines‐and‐recommendations

Many Thanks! Contact Information Mark Vander Borgh • Rachel Monschein & Erin Lee, Town of Cary Division of Water Resources (919) 743‐8423, [email protected] • Donna Hill and Neil Chernov, EPA RTP Dr. Linda Ehrlich Spirogyra Diversified Environmental Services (336) 570‐2520, [email protected]

Dr. Astrid Schnetzer North Carolina State University (919) 515‐7837, [email protected]

6