Welcome to your first training module for becoming a PMN volunteer.
This is a self guided "tour" of the basic information required to participate
and you can refer back to this recording at any time.
We will be covering three main topics:
1) what exactly harmful algal blooms are,
2) some information about our phytoplankton monitoring network,
and 3) how you will be collecting samples in the field
and analyzing your water samples
for the presence of particular types of phytoplankton,
which we refer to as our "target organisms."
The P in PMN stands for phytoplankton,
which comes from the Greek words for plant and wanderer.
Many of them can photosynthesize just like plants on land
and they are at the mercy of the movements of the currents and winds.
Phytoplankton is a general term
used to describe single-celled, photosynthetic microscopic organisms that live in aquatic environments.
Also called micro algae,
they live in mainly in the water column
where they can get enough sunlight to produce their own food,
thus forming the basis of the aquatic food chain.
The oxygen produced
is released into the water and the atmosphere,
which provides about half the oxygen that we breathe.
Phytoplankton are also important in carbon cycling
and climate control of the planet.
So life truly would not exist without phytoplankton.
It is important to keep these crucial benefits in mind
as we will be concentrating only on those
that can potentially cause harmful algal blooms.
Harmful algal blooms, or HABs,
can occur when phytoplankton grow at very rapid rates.
This can lead to obvious discoloration of the water
on small and large scales.
We prefer to use the term HAB
rather than “red tide” or “brown tide”
because there are a variety of colors a HAB can be,
and can even be harmful without discoloration of the water,
depending on the species of phytoplankton present.
Some can even bioluminesce.
Phytoplankton growth is generally seasonal,
and depends on the availability of
carbon dioxide, sunlight, and nutrients.
Phytoplankton, like land plants, require nutrients
such as nitrate, phosphate, silicate, and calcium
at various levels depending on the species.
Other factors influence phytoplankton growth rates,
including water temperature and salinity,
water depth, wind,
and what kinds of predators are grazing on them.
When excess nutrients are available
under ideal growth conditions,
blooms are more likely to appear.
Research is being done to better predict when blooms may form
to help mitigate their potential harmful effects,
but not all algal blooms are harmful!
For example, a phytoplankton called Coccolithophorids
does not produce a toxin and are generally harmless,
even though they can form blooms so large
that you can see them from space.
And there were so many in the ancient ocean
that as they died and sank to the bottom
they formed chalky sediments layers
thick enough for you to walk on top of today,
like at the Cliffs of Dover.
So what do we mean when we say "harmful"?
There are several ways
in which a bloom can be considered harmful.
One is if they cause physical damage.
For example, fish gills can become clogged
or damaged by the phytoplankton cells.
Fish respond to this damage by producing mucus,
and this can ultimately lead to death by suffocation.
When a bloom dies off,
the cells sink through the water column
and can be degraded by bacteria.
This process removes oxygen from the water,
making survival impossible for other organisms
due to the formation of dead zones
from hypoxic or anoxic conditions.
What most people often associate with a harmful algal bloom
is the production of toxins.
There are many different groups of toxins
that vary in their levels of toxicity,
and health effects can be seen in marine animals
like dolphins and sea turtles, as well as in humans.
As seen on the map, this is a global issue,
with some regions susceptible to multiple types of toxin exposure.
Not all phytoplankton are able to produce toxins;
in fact, very few are.
They must have the genes for toxin production,
and even if they HAVE the genes,
those genes may not always be turned on.
But if toxins are being made, they can travel through the food web
and concentrate in the tissues of things like shellfish and fish.
Exposure comes from either feeding on the toxic cells directly or
feeding on something ELSE that has been exposed to the toxins.
Depending on the amount of toxin ingested,
effects can range from relatively mild to lethal.
Bloom-forming toxic microalgae include three main groups of phytoplankton:
the dinoflagellates, the diatoms, and cyanobacteria.
These three types of algae produce the toxins
which cause most human and animal health problems.
The known toxin-producing species
typically do not affect human health directly,
but illness occurs through the consumption
of toxin-contaminated seafood or drinking water.
That said, sometimes toxic species don’t produce any toxin,
and the amount of toxin, again, can vary among species, as well.
These are the five human health syndromes
that are known to be directly attributed to algal toxin exposure.
The primary causative agent is shown in each,
although, in some cases,
it is not necessarily the only organism capable
of producing the different groups of toxins, shown in red.
Symptoms will vary depending on
the type of toxin that has been ingested and how much,
but in some instances, such as with saxitoxins,
it can be lethal in humans.
Unfortunately, it is not as simple as just looking at a bloom
and being able to determine if it is toxic
just by the organisms present or by discoloration of the water.
Toxins can be present even if the water is not discolored.
This could occur if the cells have lysed,
opened upon death,
and the toxins were released into the water,
but there is no color change.
Another way is if the bloom is deep enough in the water,
such that it is not visible from the surface.
As previously mentioned,
not all species have the genes to produce toxins,
and if they do, they are not always turned on.
So, the presence of a species known to produce toxins
does not always mean that toxins are present.
And for each type of toxin, there are multiple forms or congeners,
each with varying degrees of toxicity.
All of these confounding factors
make it more difficult to immediately tell if a bloom is toxic or not,
but knowing the potential for risk
based on identification of what is in the sample
is still extremely beneficial information to have
from a monitoring standpoint.
Because of the potential harmful effects of HABs,
monitoring for the presence of the organisms
that may lead to them is important for several reasons.
We know they are increasing in how often they happen,
they are beginning earlier in the year than before
and lasting longer,
and they are also expanding
in their geographic range of occurrence.
By knowing which organisms are present
and in what relative abundance,
we can use that information with other tools, like satellite imagery,
to better understand and predict where HABs might occur.
Routine monitoring can also allow us to know early on
when a bloom might be starting to form
and thus act as an early warning system
to reduce potential exposure to toxins.
So, why help PMN?
Most agencies lack the funding and resources
to monitor the entire coastline of a state,
or look at every lake and reservoir that can be impacted by HABs.
Having local citizens engaged in this effort
by being the “boots on the ground”
is crucial to increasing the capacity of monitoring
and better understanding where and when HABs occur,
thus helping NOAA and other scientists in their HAB research.
PMN is a national volunteer organization that is
trained to monitor for potentially harmful algae or cyanobacteria.
You will be trained on how to collect samples
from coastal or freshwater environments,
how to identify the organisms specific to your region,
and how to enter that data into NOAA’s database.
Based on what our volunteers find,
we can test for toxins (in a limited capacity),
alert the state or local agency
that is responsible for making management decisions about HABs
and look at long-term trends
of how HABs may be changing in how often and where they occur.
We have volunteers from a wide variety of
backgrounds and groups, both science and non-science
and our volunteers have generated
over 300,000 data points across the country since our inception.
PMN began here in Charleston, South Carolina,
back in 2001 with coastal monitoring,
and in 2015 we expanded into freshwater HAB monitoring
thanks to an interagency agreement with EPA’s Office of Water.
As a PMN volunteer,
you will be provided with all of the equipment seen here.
The only equipment we can not provide is a microscope,
but everything else needed to collect water samples,
measure salinity and temperature,
make slides and ship samples to us will be sent to you.
There is also an app called Phyto
that you can download if you have an iPhone or iPad.
This is another useful resource
to help with the identification of what’s in your sample.
There are photos of both coastal and freshwater organisms,
pronunciations of the scientific names,
and a key based on cell shape
to help narrow down your identification choices.
Unfortunately, the Android version is still being updated,
but the original, older Android version
that lacks the freshwater information may still be available.
The second part of the training will concentrate on teaching you
how to identify the target organisms for your region.
You will learn how to identify to the genus level of classification,
as the features needed to get to the species level are difficult,
or impossible to see with a regular light microscope.
If we need to determine
if you have a toxin-producing species present,
you will send us the sample and we will use our
powerful scanning electron microscope to make that determination.
Feedback will be provided,
so you will know if you have correctly identified your organisms.
This feedback is an important part of our program,
both to create a strong working relationship
between volunteers and NOAA staff,
as well as a way to ensure the quality of the data generated.
And our volunteers produce results.
They have been able to identify many blooms over the years,
and some of those were found to be toxin-producing blooms
that the state regulatory agencies were not aware of.
As previously mentioned, many blooms are harmless, but knowing
which ones have the potential to be harmful is very important!
Even our newer freshwater program
has found toxin-producing blooms.
The data generated by our volunteers is not just for PMN usage,
but other entities use our data as well,
for building models for things like drought and tide gauges
to recreational fisheries trying to avoid potential HAB areas.
This portion will cover the protocols
you will be using to collect samples
and how to go through that sample on your microscope.
You should have already viewed the training video.
that covers many of these steps.
If not, please email pmn@noaa.gov to receive a copy.
The first thing to do
is select the site that you will be sampling on a regular basis.
Where you sample is completely up to you,
but you need to be able to get there consistently,
at least every other week for a year.
It could be a dock, along a beach, or at a park,
wherever you can safely, and legally, tow your plankton net.
You need to be able to get there, again, on a consistent basis.
For picking your site,
this could be someplace where you have seen blooms,
or discolored water before;
someplace that's important to you or your community,
like a neighborhood pond, or beach;
or you might just be curious to see what's in your water.
You don't necessarily need to sample
on the exact same day every week, or at the same time.
You can schedule according to what works best for you.
But, again, you do need to be sampling at least every other week.
Once you decide what site you are going to use,
your homework is to send us your site information.
This would include a name for your site
and GPS coordinates, ideally in decimal format.
This allows us to get your site information into our data entry portal
and make you a dot on the map.
It is possible to have more than one site,
so we would need the information for each site.
Here is an overview of what PMN sampling and analysis involves.
We will go through the field sampling steps briefly,
as they were covered in the training video.
You will receive a copy of a region-specific data sheet.
This shows the target organisms you are looking for in particular.
You are not responsible for identifying everything you see in your
water sample, but you are responsible for being able to identify
the target organsims,
as these have the potential to cause harmful algal blooms.
You should bring a copy of this data sheet with you
each time you sample, in order to record your data.
The date,
time of sampling,
water and air temperatures, along with salinity,
are all required information for each sample.
Items below the dotted line on the data sheet are all optional,
like weather conditions and other measurements you can take
if you have the equipment to make them.
We will cover more details about the data sheet shortly.
So, first, you are going to do your plankton tow.
It is important that there is no air in your net or the bottle.
You'll dunk your net until the bottle is full of water.
Then, let it sink vertically as deep as you can
without hitting the bottom.
This depth can vary with tides and could be several feet,
or just a few inches deeper than the mouth of the net.
After your vertical drop,
you'll bring the net back up to about a foot below the surface,
and you'll begin your three-minute timer for the horizontal tow.
Try to avoid getting anything in the net that is not water,
like sea grass or trash,
and avoid towing nets through visible oil or gas slicks.
Do not let the net bump into pilings that may have mussels
or barnacle growth, as this may tear the net.
Please refer to the training video for more details on the net tow
and how to concentrate your sample at the end.
You will also need to record air and water temperatures.
You can tie a line on to the thermometer
and tie it off to a cleat or railing
and throw it into the water while you do your net tow.
You do want to give the thermometer
time to acclimate to the water temperature,
and take your reading quickly after removal from the water.
Do not let the thermometer lay on the ground or dock
when taking air temperature, as this will not be accurate.
You can also tie it to a railing
to hang while you do your salinity reading,
or hold it at the top of the thermometer
to get an accurate air temperature.
You will also be using a refractometer to take a salinity reading.
Be sure to have the platform flooded with water before closing
the cover, and point it towards the brightest part of the sky
to get good contrast between the blue and white backgrounds.
You will take the reading where those two backgrounds meet,
using the values on the right-hand side.
The last thing to do is collect the whole water samples
in the 1 L and 30 mL bottles.
All you have to do is fill them up.
While at your site,
be sure to take a look around and observe the conditions,
including, optional weather details as seen on the data sheet,
whether the water is discolored or smells bad, etc.
The entire sampling process
should take about 10–15 minutes total.
You do want to take care of your equipment,
so please be sure to give everything
a good freshwater rinse when you return home.
Do not use soap on the plankton net
and be sure to hang it up to dry,
as opposed to letting it lie in a sink or a tub.
Ideally, you want to look at your sample as soon as possible,
and definitely within 6–8 hours of collection.
You will take your sample from the 125 mL bottle
that was attached to the net.
Gently re-suspend the bottle.
Then, collect some water with your pipette,
and add TWO drops to your gridded slide.
Place a cover slip on top, as shown,
placing one edge of the cover slip on top of the slide, and
lowering the opposite edge as low as you can before letting it go.
This will reduce the amount of air bubbles
trapped under the cover slip.
Then, you’re ready to do your analysis.
Each gridded slide is an 8x8 gridded square,
and you will be looking through all 64 grids for your analysis.
The objective is to look for any target organisms present
and determine the relative abundance of each.
You can start in any corner,
using your 10x objective.
Move around in that grid, and look for things that might be targets.
If you see one, focus it in the exact middle of your field of view,
move up to your 40x objective, and make your identification.
If it is a target organism, this is when you would take a photo.
Once you’ve made your ID then go back down to the 10X objective
and continue viewing through that grid
until it has been completely viewed.
Then, move to the next grid, and begin again.
You will go through all 64 grids using this manner
and using a lawn-mower method, going row by row,
to ensure every square is viewed.
You will not need to count all cells,
but you do want to keep track
of how often you observe each target organism.
This will be covered in more detail shortly.
Taking photos of target organisms is crucial for our program.
Any time you identify something as a target organism,
we will need one or two representative photos,
so that we can confirm your identification.
This way we will ensure that the data that get entered
into our database are correct and are validated.
You will be given feedback on your IDs,
so you will know how you’re doing.
We do not need a photo of EVERY single target cell you may see,
but, again, at least one or two representatives
of each target group that are present in your sample.
Photos should be taken at 400X total magnification,
which is when you are using your 40X objective.
If you identify something that is a non-target organism,
meaning a more common type of phytoplankton
that is not necessarily harmful,
take a photograph of that, as well.
If you are not sure what something is,
take a photograph, and we will try to ID it for you.
The goal is for YOU to make the ID, and for us to confirm it.
The video I'm about to show
lets you know how to use your cell phone camera
to take photographs through a microscope.
The most important part of this process is stability,
and it does take some practice.
So,
this is one method to take a photograph
if you don't have a dedicated camera
already connected to your microscope.
Stability is the most important part of this process.
So, using the three fingers on your left hand,
brace your hand against the eyepiece.
Hold your phone between your thumb and first finger.
Hold the other side with your opposite hand
so the thumb is free to click your camera button.
Starting a few inches away,
find the center of the circle of white light with the camera
and move the phone closer
until the image in the microscope comes into focus.
Use your thumb to take the photograph.
To get rid of the black edges,
you can zoom in slightly first on your camera,
then, again, stabilize with your left hand,
focus the white light until the object comes into focus
and there should be no black edge
when you take your photograph.
This does take practice.
So, keep trying!
Don't get frustrated if it doesn't work for you right away.
You do not want to email your photos to us
directly from your cell phone
if that is the method you use to take pictures.
Please upload all photos to your computer first.
When you're finished analyzing your slide,
you're going to label each photo with your identification.
Once all the photos are labeled,
you're going to complile all the photographs into one file.
This can be a Word document, a PDF, or a PowerPoint presentation.
Your PMN ID, your site name, and the sample date
should also be included.
This could be put into the file name,
or at the beginning of the file,
as you see in the example on the right.
You will email this file to PMN@noaa.gov
and this is what you would receive feedback on.
Please be patient with us to receive your feedback,
but it is, usually, within one week.
If you are just starting out,
please submit your photos before you enter your data
to reduce corrections that may need to be made to the database.
So, by submitting your photos first, and getting the feedback,
you will be entering the corrected data.
Again, you will receive a data sheet specific to your region,
with your target organisms listed at the top.
When you are done looking through the entire slide,
you need to have an answer of NO, YES, or ELEVATED
for each of the listed targets for your region.
NO means that you did not see any of that particular target organism.
YES means you did see at least one cell
and maybe up to a few cells over the entirety of the slide.
ELEVATED means that you see multiple cells of that target in a given grid,
and in a majority of the grids on the slide,
meaning this is your dominant organism.
Again, there’s no need to count the cells,
but we are asking for a relative abundance.
Are there NONE? Are there SOME? Are there LOTS?
Here are some examples to show you.
NO means exactly that: No cells for that target were seen anywhere on the slide.
YES means that you are seeing some of that target scattered across your slide.
For example, if you see just one Dinophysis, which is this cell here,
and that is the only one you see, it is a YES.
Now, if you see a few Dinophysis cells here and there,
maybe some here, maybe some over there, one or two over there,
but, overall, they are found in less than 60% of the 64 grids on your slide,
then that is still a YES.
Where to draw the line between a YES and an ELEVATED is subjective,
but it does get easier with practice.
Now, the lower end of ELEVATED might start to look something like this.
In each of these grids,
so we’re assuming each of these photos represents one grid on your slide,
there are multiple cells of the same target organsim.
If you start to see this in 60% or more of the grids overall,
meaning you are seeing multiple cells of that target in the majority of your grid,
then that would be considered a lot, or ELEVATED.
Sometimes this is easier to determine.
Here, you see a common phytoplankton called Rhizosolenia,
which look like glass cigars.
Rhizosolenia is dominating this particular grid,
and, presumably, the rest of the slide as well.
There are still other species of phytoplankton visible,
but it is clearly dominated by the Rhizosolenia.
This would definitely fall into the elevated category.
And if there is one species that seems to cover the entire slide,
then that is most certainly elevated and a bloom.
One usually sees discoloration of the water sample with concentrations of cells like those shown here.
Hopefully, this gives you a visual idea of how to bin relative abundances during your sample analysis.
Remember, you must have an answer of no, yes, or elevated for each target on your data sheet.
These categories are not describing the sample as a whole.
The last thing to do, after you have analyzed your slide, is to enter your data.
This is another very important part of our program.
Once you provide a site name and GPS position,
you’ll be given a PMN ID,
which will correspond to the region that you are in.
To enter your data, you’ll go to our homepage, and click the “submit data” button.
You will then choose your desiganted region,
and it will bring you to a Google form to fill out.
You will type in your email,
find your region, your PMN ID, and site name,
and then fill in all the required information from your data sheet.
For optional information, like the weather, or non-target organisms,
those fields are pre-filled as “not applicable.”
You may fill them in, if you would like.
Photos are not uploaded here, but are emailed separately to PMN@noaa.gov
So, what do you do with your bottles once you are done?
That depends on what you have found in your sample.
You’ll notice on your data sheet,
that there are symbols in your “no,” “yes,” and “elevated” columns.
These symbols will tell you which bottles you should preserve and save for shipment to us here at NOAA.
The description of what each symbol means is found on your data sheet.
We will first go over how to preserve samples, and then how often to ship them.
If a symbol tells you to preserve a sample,
you will add something called Lugol’s solution, which will also be provided to you.
This kills the cells, but keeps them intact for further viewing.
This solution is harmless, but it will stain your skin or clothing, so please be careful when using it.
If you are asked to preserve your 30-mL bottle,
you will fill the provided pipette with Lugol’s up to the first mark,
That is about 0.5 mL. and add that to the bottle.
You will add that to your 30-mL bottle.
If you are asked to preserve your 125-mL net tow bottle,
you will fill it to the 2nd mark with Lugol’s,
and add that to your 125-mL bottle.
A preserved bottle should have the color of strong, iced tea.
If your sample is very dense with phytoplankton,
you may need to add some additional Lugol’s to achieve that color.
You will never add Lugol’s to the 1-Liter bottle!
Once you’re done, you’ll add black, electrical tape to seal the lid shut,
add a filled-out sample label,
and put your bottles in a Ziploc baggie.
You will want to store your preserved bottles and your Lugol’s solution
in a cool, dark place—like a cabinet, or a drawer—
until it’s time to ship the bottles to PMN, according to the calendar.
We will go through some examples, now.
So, if after looking through your entire slide
you do not see any target organisms, you will have all NOs.
The symbols are all circles,
so you do not need to save ANY of your bottles for shipment.
You will empty and rinse all three, and save them to reuse again.
Remember, though, even if you do not see any targets,
you still need to enter the data for that sample!
It’s very important that you enter data for each and every sample,
even if no targets are present.
For this example, notice that there are circles in the YES column, as well.
But, if at the end you still have all circles, the same applies:
empty and reuse all three bottles. There is no need to save them.
If you have a triangle in either the YES or ELEVATED column,
you will preserve both the 30-mL and the 125-mL net tow bottle.
Once they’re preserved, put them in your drawer, or cabinet,
and wait to ship them until the calendar says to.
If you have any diamonds in the YES or ELEVATED column,
you will only save the 30-mL bottle.
Any bottle you are not asked to save, you will rinse and reuse.
This means that 99.99% of the time you will be entering the 1-Liter bottle.
You will be receiving a shipping calendar.
This will tell you when to send all of your saved sample bottles over the course of two months.
For example, if you start sampling in July,
then you would send all of your preserved bottles in September,
according to your region as noted at the bottom.
You, again, will place the bottles in a Ziploc bag.
You’ll place all of those in a white, plastic envelope that is provided,
and you will use the appropriate shipping label.
Most of the time, you will be using this brown, ground shipping label.
You just need to fill out the areas marked in red with your information,
and drop the package off at any UPS store, or call for UPS pick up.
You should never be charged to ship your samples with these labels!
Now, there may be a time when you have a square for your symbol.
These are when you see elevated levels of a target
that can potentially cause one of the five human health syndromes mentioned earlier.
These are the samples that we want to have shipped IMMEDIATELY to us.
You’ll preserve the two smaller bottles.
You’ll, again, NOT add Lugol’s to the 1-Liter,
but you will send the 1-Liter bottle, as well.
And all three bottles need to be sent overnight.
Only in this instance will you use the red, overnight UPS label.
You will fill it out the same as the ground label,
but please remember to either take the “shipper’s” carbon copy,
or take a photograph of the label, so you have a copy of the tracking number.
You will email us to let us know a sample is coming overnight,
so we can prepare to work it up upon arrival.
And you will also provide us with that tracking number, as well.
So, when the sample bottles arrive,
we will first look at the 125-mL bottle
to look at the sample under the scanning electron microscope to get the species identification.
If it turns out to be a known toxin producer,
we will then use the 1-Liter bottle for toxin testing.
This is why it’s important NOT to add Lugol’s to the 1-Liter bottle,
as that color change will interfere with the assay.
The 30-mL bottle is used to get an actual cell count.
Again, any bottles you are not asked to save for shipment will be reused.
And remember to always enter data for every sample,
even if you do not see any targets.
So, here is just a quick review of the protocols we have covered here today.
If you have any further questions about the protocols,
please don’t hesitate to email us.
Now that you’ve completed Part 1 of the training,
you must first complete a review of the information.
This is done using a program called Menti.
Your answers are anonymous,
but it gives us an idea of how well we are getting our information across to our volunteers.
Please email pmn@noaa.gov
and request the special code to use for the review.
Once you have completed that review quiz,
you can then request to set up the second part of training.
Part 2 will be done live via webinar, and according to your specific region.
Possible dates will be determined on a rolling basis.
When Part 2 is complete,
your supplies will then be shipped to you,
and you can begin sampling for PMN.
Again, if you have any questions,
you can email us at pmn@noaa.gov
or Jennifer.Maucher@noaa.gov
Thank you!