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PhytoplanktonMonitoring Network

Freshwater CyanoHAB Program

Promoting a better understanding of Cyano Harmful Algal Blooms by way of volunteer monitoring.

Freshwater

NOAA Marine Biotoxins ProgramJen Maucher Fuquay, PMN Coordinator

(PMN)

Cyanobacterial Harmful Algal Blooms (cyanoHABs)

Cyanobacteria, formerly called blue-green algae, are a type ofphotosynthetic bacteria.

Beneficial in that they can produceoxygen for the water andatmosphere.

When environmental conditions areideal, cyanos can grow rapidly, or‘bloom’, forming thick surface scumlayers.

Cyanobacterial Harmful Algal Blooms (cyanoHABs)SOME species are potentiallyharmful, and can produce toxins thatcan impact drinking and recreationalwaters.

Can affect wildlife, pets and humans

Paul S. Sarbanes Environmental Restoration Site at Poplar Island Project

• Block the sun– Kills other aquatic plants, reducing oxygen production

• Remove oxygen from water when bloom dies off– Kills other organisms like fish

• Produce Toxins– Can cause illness in animals and humans

Harmful Effects of CyanoHABs

Cyanobacteria have been linked to human and animal illnesses around the world, including North and South America, Africa, Australia, Europe, Scandinavia, and China.

Cyano Harmful Algal Blooms (cyanoHABs)

Credits: NOAA

Lure Bar & Grill, Lake Petenwell, WI

Mounting evidence indicates globalclimate changes support increasedfrequency and geographic extent ofHABs.

Each year the desire to live at lakesideand the reliance on large surface watersfor recreation and drinking water putsmore people and animals at risk forexposure to HABs and the toxins theycan produce.

Presenter

Presentation Notes

Do extremely well in high temperatures

Problem: Very few government or private institutionshave the capacity or capability to monitor thousandsof lakes (and reservoirs), from the Great Lakes to smallresidential lakes annually impacted by HABs.

Why the PMN?

Solution: Engage local citizens in environmental monitoring of potentially harmful cyanobacteria to aid NOAA scientists and others in their research.

Train citizen scientists to:• identify algal species• analyze water samples for

HAB toxins• identify temporal and

geographic HAB trends

• PMN started in 2001 as part of Marine Biotoxins Program in Charleston, SC

• Over 100 active sites in 13 coastal states

PHYTOPLANKTON MONITORING NETWORKNATIONAL CENTERS FOR COASTAL OCEAN SCIENCE

Science Serving Coastal Communities



• CyanoHAB monitoring started in 2015 as part of an EPA Office of Water grant

• 22 active sites in 10 states

Cherry Creek Reservoir, Denver, CO June 2016Dolichospermum bloom with low MC levels

Train citizen scientists to:• Collect samples on weekly or bi-

weekly basis• Identify potential harmful algal

species

NOAA scientists can then:• analyze water samples for HAB

toxins• Together can identify temporal

and geographic HAB trends



Monitoring Benefits

• Allows for an ‘early warning system’ – e.g. Can close shellfish beds/recreational waters and help prevent

people from getting sick

• Monitor and maintain an extended survey area along coastal & fresh water bodies throughout the year

• Create a comprehensive list of harmful algal/cyano species inhabiting marine and fresh waters (establish baseline)

• Identify general trends where HABs are more likely to occur • Promote an increased awareness and education to the public

on HABs • Create a working relationship between volunteers and

researchers

• Entered into a partnership tocreate a freshwater citizenscience program to monitorCyanoHABs

-FY15 included a pilot program inEPA Region 5 and Region 8

• Used previously established PMN methods with modifications for freshwater habitats.

2015 PMN and EPA

FED

ERAL

& IN

TERN

ATIO

NAL

MAN

DATE

SFO

RHA

BS

The Harmful Algal Bloom and Hypoxia Research and Control Act

(2014/2010/2004/1998)

authorizes funding for research on HABS and hypoxia to advance scientificunderstanding and the ability to detect, assess, predict, control, and mitigate events.

PHYTOPLANKTONMONITORING NETWORK

NATIONAL CENTERS FOR COASTAL OCEAN SCIENCEScience Serving Coastal Communities


Training & Protocols


Freshwater

(PMN)

Tools for cyanoHAB training & identification

WebEx training sessions– Done remotely to make

training more convenient when on-site is not possible

– Allows NOAA scientists to observe real samples online with volunteers

Phyto app version 2- coming soon!

http://youtu.be/ItzxoB06De0

-Will include freshwater species!-More pictures!-More pronunciations!-Can enter data from phone or ipad!

Developed by PMN volunteer Shawn Gano to assist with and improve volunteer’s identification skills of marine algae & cyanos

http://youtu.be/ItzxoB06De0

Phytoplankton Monitoring Network

1) Collect sample at least once every two weeks during the sampling season (Ice Out – Ice In)

2) Analyze sample identifying target algae3) Take digital pictures to send into the PMN4) Input data into the PMN database5) Preserve sample and ship to PMN when water is

visibly green

Volunteer Requirements:

1. Collect Sample

Step 1:Collect Whole Water Live Surface Samples

125 ml & 1 L

• 125 mL and 1 L bottles will be provided to volunteers• Wear gloves when sampling• Dip bottles to collect samples• Collect air & water temperatures, optional weather data and note any other observations (water color, odor, etc)

Freshwater Phytoplankton Monitoring Network

1a. Preserve SampleIf samples cannot be analyzed within 12 hrs, you can preserve the 125mL sample in the field. Lugol’s solution:

IodinePotassium iodineGlacial acetic acid

1. Add 2.5 mL to 125mL bottle2. NEVER add Lugol’s to live 1 Liter bottle!3. Keep ALL sample bottles out of direct

sunlight and heat. 1. Can bring small cooler with ice pack2. Do not need to be frozen


125mL

X

Freshwater Phytoplankton Monitoring Network Volunteer Requirements:

2) Analyze sample identifying target algae

Credit: NOAA


1) Gently mix 125 mL bottle by inverting 2x2) Take sample from middle of bottle & add

2 drops to gridded slide

3) Scan entire slide using 10X objective• Use ‘lawn mower method’• Go up to 40X objective when needed to ID

4) Identify any target species that may be present

2. Sample Analysis

FPMN Project Impacts:Important to Remember...

You are only responsible for reporting on the 5 target freshwater algae species!

There may be lots of other “stuff” in the water - feel free to investigate,

explore and have fun!

Keep in mind that sometimes you may not see any target species!


Take digital pictures and email to [email protected]

Credit: NOAA, Jeff Paternoster

Credit: Dave Florenzano Credit: Dave Florenzano

Credit: Dave FlorenzanoCredit: Dave Florenzano

Credit: Rob Dryke

Image: www.microscopeworld.com

3. Photograph sample

Take multiple pictures per sample.Be sure to label each photo with

tentative ID and the total magnification used!

e.g. Microcystis_400XUnknown1_@100X

(Bonus points for size bars)


4. Input data

*Log on to http://products.coastalscience.noaa.gov/pmn/screening.aspx

*Choose FRESHWATER

*Enter ALL required data

Image: NOAA PMN

ID will be assigned

Type in site name

Enter in 24-hr format

Enter as MM/DD/YY

Enter to nearest 1ppt

Default is “No”

http://products.coastalscience.noaa.gov/pmn/screening.aspx


4. Input data

*Other section is completely OPTIONAL!

Select from list

Enter any data

Options: No, Yes, Elevated

Can add name of dominant species if known, but it is not required

Type special comments or observations like color, odor, etc

Final step is to “submit”


• If your state/organization already has toxin testing capabilities, you do not need to send us a sample

• Follow their shipping protocols– We would appreciate the sharing of

results to keep track of toxic cyanoHAB events

• If you do NOT have any access to microcystins testing, then………

What to do if water is GREEN:

Sample bottle from August 2015 Microcystis bloom in Lake Erie (credit: Tom Green)


• Pictures will be used to initially ID potential target species

• PMN may then ask to have 1 L sample bottle sent for possible microcystins toxin analysis via ELISA.

• Must be sent ASAP!

What to do if water is GREEN:

Sample bottle from August 2015 Microcystis bloom in Lake Erie (credit: Tom Green)

Dolichospermum, Microcystis, Planktothixare all potential microcystins producers


5. Ship samples1. Add 2.5 mL to

125mL bottle2. NEVER add

Lugol’s to live 1 Liter bottle!

3. Keep ALL sample bottles out of direct sunlight and heat.

X


5. Ship samples• Add completed

sample labels to 1 L bottle & 125 mL bottles

• Wrap lid with tape and put in plastic envelope

• Fill out red UPS label for overnight shipment


5. Ship samples

• Samples can be shipped on blue ice if shipping will take longer than 24 hrs from remote locations

1. Collect whole water live samples (125mL & 1L)- wear gloves!

2. TEMPs- air and water

3. Optional info-- Observe and record surrounding conditions (wind, tide, weather)

- Other important observations: discoloration and/or smell of water

Field Sampling – Summary

Phytoplankton Identification Steps:

1. Prepare gridded slide- Mix the sample gently to re-suspend algae in the bottle- Place two drops of the sample on the gridded slide- Place a cover slip on the slide- Place the prepared slide on the microscope

2. SCAN through slide one time- Use 10X objective to scan through each grid- use 40X objective to identify individual organisms- Use ‘lawn mower’ method to ensure the entire slide has been viewed.

3. PHOTOGRAPH target species if present- Take multiple photos to get best chance of proper ID-Label each photo with target species ID and total magnification

Lab Analysis – Summary

Data Entry- Summary

1. Access the database using the NOAA PMN website2. Enter your unique log in ID, sample site name & date

• If you have multiple sites, you will use same log in ID for each site• Each site name must be entered separately into the database.

3. Enter data4. Submit!


Cyanobacteria


Freshwater

(PMN)

Greek Cyano = dark blue bacteri = bacteria

Photosynthetic bacteria – prokaryotic cells Not true algae, although we still call them HABs

CyanoHABs = Cyanobacterial Harmful Algal Bloom

Live in freshwater, brackish, and marine waters Unicellular to multicellular, some Colonial

Toxic Cyanobacteria in Water: A guide to their public health consequences, monitoring, and management, World Health Organization, 1999

Cyanobacteria Basics

Terminology:

Akinete(s)- thick walled resting spore

Heterocyte(s)- site of nitrogen fixation; also thick walled

Trichome(s)- a filament (rows) of cells which are connected

FPMN Project Impacts:Important to Remember...

Algae and cyanobacteria are a natural part of the water way. NOT all are

“bad.”

However, some can produce toxins.The PMN targets 5 freshwater cyanos that can be harmful, all of which are potential

toxin producers.

Microcystis spp.

Aphanizomenon spp.

aka Oscillatoria spp.

Cylindrospermopsis spp.Dolichospermum spp.

Target Freshwater Algae


aka Anabaena spp.

Planktothrix spp.

Toxins Produced Type of Toxin

Target Organ Onset of symptoms

Anatoxin-a

Neurotoxins Nervous SystemLabored breathing, convulsions, numbness, paralysis and deathDog deaths caused by Anatoxin-a

Minutes to hoursSaxitoxins

Microcystins HepatotoxinLiverGI symptoms, elevated liver enzymes in blood, death of cells, destruction of blood vessels

Hours to days

Cylindrospermopsins HepatotoxinLiver and KidneysSymptoms like food poisoning/Pos. kidney failure

Hours to days

CyanoToxins

nono

no

no

noyes

yes yes

yes

yes

no

Cannot use taxonomy to predict toxicity

Slide blatantly stolen from Barry Rosen @USGS

Freshwater Morphology~ 5 Target Species

NOAA Marine Biotoxins ProgramDr. Steve Morton, Principal Investigator



Microcystis spp.

Aphanizomenon spp.

aka Oscillatoria spp.

Cylindrospermopsis spp.Dolichospermum spp.

Target Freshwater Algae


aka Anabaena spp.

Planktothrix spp.

Fres

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Dolichospermum spp.N-Fixer

• Filamentous• Unbranched & untapered• Trichomes usually solitary• No sheath*• Can be straight, curved or spirally

coiled• Large temp and salinity ranges

Credit: GreenWater Laboratories/CyanoLab

Terminology:

Akinete(s)- thick walled resting spore

Heterocyte(s)- site of nitrogen fixation; also thick walled but clear

Trichome(s)- a filament (rows) of cells which are connected


Anabaena has now been re-classified by some as Dolichospermum

Things to look for:A. Cells rounded or barrel shaped with aerotopesB. Heterocytes are intercalaryC. Akinetes are intercalaryD. Cells constricted at cross walls

DOLICHOSPERMUMB

C

A

Photo: © A. Ballot D

DOLICHOSPERMUM

Fres

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Dolichospermum (Anabaena) spp.N-Fixer

Akinetes known to survive more than 64 years in sediment



Akineteheterocyst

Cred

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Dolichospermum can be confused with Nostoc, but Nostoc trichomes form colonies

Cred

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, USG

S

Cred

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Andrew Chapman- Greenwater Laboratory

Fres

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Aphanizomenon spp.N-Fixer

• Filamentous, straight, unbranched trichomes

• Tapered at both ends• No sheath• Trichomes arranged in

parallel layers.• Has heterocysts and forms

akinetes• Can form winter & summer

blooms

Approximately 15 known species

Akinetes known to survive more than 18 years in sediment


B

CD

A

D

Photo: © A. Ballot

Things to look for:A. Aerotopes (facultative)

B. Heterocytes are intercalary (facultative)

C. Akinetes usually cylindrical and intercalary

D. Terminal ends are elongated and may be “empty” looking

Aphanizomenon

Fres

hwat

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yano

bact

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Aphanizomenon spp. Can be confused with :

Anabaena-Can sometimes be found in bundles

but cells are more rounded than Aphanizomenon.

Planktothrix-Can look similar to individual Aphan. trichomes, especially in absence of

heterocytes.

Cred

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S

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‘lawn clippings”

Fres

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Aphanizomenon bloom

Will be filamentous

Credit: GreenWater Laboratories/CyanoLabCredit: Dr Barry Rosen, USGSAphanizomenon Flos-Aquae Credit: Klamath Valley Botanicals

Fres

hwat

er C

yano

bact

eria

Cylindrospermopsis spp.N-Fixer

• Filamentous, unbranched• Trichomes are straight,

curved or coiled; solitary• No sheath• Heterocysts always

terminal!• Akinetes form behind or

slightly distant from heterocysts (gives asymmetric appearance



A

B

B BC

A

Things to look for:A. Cells cylindrical with aerotopes

B. Heterocytes (when present) are always terminal at one or both ends

C. Akinetes (when present) usually 1-3 cells back from heterocytes

D. Terminal cells attenuate (get narrower)

-Difficult to ID when lacking heterocytes

CYLINDROSPERMOPSIS

D

Fres

hwat

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yano

bact

eria

Cylindrospermopsis spp.

Approximately ?? known speciesCan be confused with :

Raphidiopsis curvata-has tapered terminal cells, NO

heterocysts, and any akinetes will always be near middle of chain,

never at end

Cylindrospermum spp.-Also has terminal heterocyst; benthic

or epiphytic instead of planktonic

Cout

e &

Bou

vy (2

004)

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reen

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Fres

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Microcystis spp.

• Colonial• Unicellular but held

together by snotty sheath• Colonies are irregular,

cloud-like (3D) with hollow spaces

• Buoyant due to gas vesicles• Smells bad!• Zebra mussels selectively

reject Microcystis cells

Approximately 25 known species

Credit: All after Entwisle et. Al (1977)

Held together by mucilaginous sheath

B C

A

Things to look for:A. Rounded cells with aerotopesCells in colony may be

A. loosely associatedB. clathrateC. densely packed

Mucilage can vary in thickness

Microcystis

Fres

hwat

er C

yano

bact

eria

Microcystis spp.

Can be confused with :

Woronichinia sp.

Woronichinia makes a hollow ball of cells

Image by Jason

Oyadom

ari

Cred

it: D

r Bar

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osen

, USG

S

Fres

hwat

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yano

bact

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Microcystis bloom

Will NOT be filamentous and will be quite stinky

Image by Jason

Oyadom

ari

Fres

hwat

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yano

bact

eria

Planktothrix morphotype

• Filamentous, unbranched

• Trichomes cylindrical, straight or slightly wavy

• No sheath• No heterocysts • No akinetes• Motile with gliding

oscillations

More than 100 known species

Cred

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reen

Wat

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Cyan

oLab

Credit: Ohio University

Dillard, G. (1999).

Formerly classified as Oscillatoria

Cred

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OAA

AB

B C

A

Things to look for:A. Cells cylindrical; mostly wider than long

B. LOTS of aerotopes throughout cells

C. Terminal cells rounded

- No heterocytes (not a N2 fixer)

- No akinetes

PLANKTOTHRIX

B CA

Fres

hwat

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yano

bact

eria

Planktothrix morphotype.

Cred

it: G

reen

Wat

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abor

ator

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Cyan

oLab

Can be confused with :

Lyngbya-Has a visible sheath

Dr. Barry Rosen, USGS

Fres

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yano

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Planktothrix bloom

Cred

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reen

Wat

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Cyan

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Credit: Ohio Universityht

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FilamentousCan be red or green depending on species

Planktothrix rubescens

Funding partners

Questions?Please Contact:

Jen Maucher [email protected]



mailto:[email protected]

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