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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.
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
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
PHYTOPLANKTON MONITORING NETWORKNATIONAL CENTERS FOR COASTAL OCEAN SCIENCE
Science Serving Coastal Communities
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
PhytoplanktonMonitoring Network
Training & Protocols
Promoting a better understanding of Cyano Harmful Algal Blooms by way of volunteer monitoring.
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
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
Freshwater Phytoplankton Monitoring Network
125mL
X
Freshwater Phytoplankton Monitoring Network Volunteer Requirements:
2) Analyze sample identifying target algae
Credit: NOAA
Freshwater Phytoplankton Monitoring Network
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!
Freshwater Phytoplankton Monitoring Network
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)
Freshwater Phytoplankton Monitoring Network
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”
Freshwater Phytoplankton Monitoring Network
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”
Freshwater Phytoplankton Monitoring Network
• 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)
Freshwater Phytoplankton Monitoring Network
• 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
Freshwater Phytoplankton Monitoring Network
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
Freshwater Phytoplankton Monitoring Network
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
Freshwater Phytoplankton Monitoring Network
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!
PhytoplanktonMonitoring Network
Cyanobacteria
Promoting a better understanding of Cyano Harmful Algal Blooms by way of volunteer monitoring.
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
Toxic Cyanobacteria in Water: A guide to their public health consequences, monitoring, and management, World Health Organization, 1999
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
PhytoplanktonMonitoring Network
Promoting a better understanding of Cyano Harmful Algal Blooms by way of volunteer monitoring.
Microcystis spp.
Aphanizomenon spp.
aka Oscillatoria spp.
Cylindrospermopsis spp.Dolichospermum spp.
Target Freshwater Algae
Toxic Cyanobacteria in Water: A guide to their public health consequences, monitoring, and management, World Health Organization, 1999
aka Anabaena spp.
Planktothrix spp.
Fres
hwat
er C
yano
bact
eria
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
Credit: GreenWater Laboratories/CyanoLab
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
hwat
er C
yano
bact
eria
Dolichospermum (Anabaena) spp.N-Fixer
Akinetes known to survive more than 64 years in sediment
Credit: GreenWater Laboratories/CyanoLab
Credit: GreenWater Laboratories/CyanoLab
Akineteheterocyst
Cred
it: D
rBar
ry R
osen
, USG
S
Dolichospermum can be confused with Nostoc, but Nostoc trichomes form colonies
Cred
it: D
rBar
ry R
osen
, USG
S
Cred
it: D
rBar
ry R
osen
, USG
S
Andrew Chapman- Greenwater Laboratory
Fres
hwat
er C
yano
bact
eria
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
Credit: GreenWater Laboratories/CyanoLab
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
er C
yano
bact
eria
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
it: D
r Bar
ry R
osen
, USG
S
Cred
it: h
ttp:
//cf
b.un
h.ed
u/ph
ycok
ey
‘lawn clippings”
Fres
hwat
er C
yano
bact
eria
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
Credit: GreenWater Laboratories/CyanoLab
Credit: GreenWater Laboratories/CyanoLab
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
er C
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)
Cred
it: D
r Bar
ry R
osen
, USG
S
Cred
it: D
rBar
ry R
osen
, USG
S
Cred
it: A
ndre
w C
hapm
an, G
reen
wat
er L
abor
ator
y
Fres
hwat
er C
yano
bact
eria
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
ry R
osen
, USG
S
Fres
hwat
er C
yano
bact
eria
Microcystis bloom
Will NOT be filamentous and will be quite stinky
Image by Jason
Oyadom
ari
Fres
hwat
er C
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
it: G
reen
Wat
er L
abor
ator
ies/
Cyan
oLab
Credit: Ohio University
Dillard, G. (1999).
Formerly classified as Oscillatoria
Cred
it: N
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
er C
yano
bact
eria
Planktothrix morphotype.
Cred
it: G
reen
Wat
er L
abor
ator
ies/
Cyan
oLab
Can be confused with :
Lyngbya-Has a visible sheath
Dr. Barry Rosen, USGS
Fres
hwat
er C
yano
bact
eria
Planktothrix bloom
Cred
it: G
reen
Wat
er L
abor
ator
ies/
Cyan
oLab
Credit: Ohio Universityht
tp:/
/cfb
.unh
.edu
/
FilamentousCan be red or green depending on species
Planktothrix rubescens
Funding partners
Questions?Please Contact:
Jen Maucher [email protected]
PhytoplanktonMonitoring Network
Promoting a better understanding of Cyano Harmful Algal Blooms by way of volunteer monitoring.