blue water task force procedures manual
TRANSCRIPT
Page | 1
Surfrider Foundation SLO Chapter
“Keeping It Blue” Project
Blue Water Task Force
Procedures manual
For
Water Sampling and Lab Testing
Compiled* by Niel Dilworth
August 2020
COVID-19 ed.
*This is an abbreviated description of our sampling and lab testing procedures.
For complete information, please refer to the Water Sampling Instructions and
BWTF Lab Instructions, which are available at the following link:
www.bwtf.surfrider.org/resources
Page | 2
ONE: COLLECTING THE SAMPLE
1. Go to the sample collection site.
2. Depending on the collection method, prepare one
sample baggy or 100ml vial for each site where a
sample is to be collected.
3. Write the time, date, and location of the sample on
the baggy or the cap of the 100 ml. vial.
4. Take the baggy/vial to the location where the sample
is to be taken.
Page | 3
5. If you are using a baggy to collect the sample –
At the location, carefully open
the baggy by pulling the two
tabs apart. Do not touch the
mouth of the baggy.
6. Walk at least knee deep into the water. Hold the
sample baggy by the tabs and submerge the baggy at
least 6” (six inches). Fill the baggy to the water line
remembering not to touch the mouth or the inside of
the baggy.
7. Close the baggy carefully without touching the mouth
of the baggy.
Page | 4
8. If you are using a 100ml. vial to collect the sample –
Write the location and time on the bottle cap.
9. Place the bottle in the dipper making sure that the
rubber band is up on the shoulders of the vial. This will
keep the vial from popping out of the dipper when it
hits the water. Remove the bottle cap and place it in a
clean location.
10. Lower the dipper into the water with enough force to
submerge the vial. Once the vial is filled retrieve it and
recap the vial.
Take the sample(s) back to the lab immediately and
prepare the sample for testing. NOTE: If you cannot
immediately test the sample, you must refrigerate it or
put it on ice in a cooler, but in no case for longer than 6
(six) hours.
Page | 5
TWO: TESTING THE SAMPLE
COVID-19 REQUIREMENTS –
All volunteers must wear a mask or face covering at all
times when they are in the aquarium building and maintain
6’ social distancing with other BWTF volunteers and
aquarium staff.
Overview of Lab Area
The Quanti-Tray Sealer
The Incubator
Page | 6
1. If you need to unlock the exterior door to enter the
aquarium, upon entering check to see if the alarm is
armed. If it is not armed proceed to Step 2. If it is
armed (the system will be beeping) enter the alarm
code on the keypad and press ENT., NOT COMMAND.
If you have trouble with the alarm system or if the
alarm sounds, call Mace Security at their number on
the keypad. Explain who you are and that you are
having trouble with the alarm and ask for help to reset
the system.
2. Enter your name, date and time in the Log in the front
of the binder that holds the location sample sheets.
3. If they are not already turned on, turn on the tray
sealer and the incubator and confirm that the incubator
is set to the correct temperature.
Page | 7
4. In the Sample Logs in the binder write in the time,
date, and location of each of your samples, as well as
your name, the tide and the weather.
5. Sterilize the workspace and sink area: take a paper
towel and clean the surfaces with the 10% Clorox
bleach solution. Let dry.
6. Put on clean lab gloves.
7. Pull out these items and place on the sink area: THE SAMPLE(S) THAT YOU COLLECTED
THE DISTILLED WATER THE PIPETTE TOOL (The Extractor) and for each sample: A 100 ml PLASTIC SAMPLE BOTTLE A sterile PLASTIC PIPETTE AN ENTEROLERT PACKET FROM THE REFRIGERATOR A QUANTI-TRAY®
Page | 8
8. Label the Quanti-Tray and the 100 ml plastic bottle
with your sample’s location, date, time.
9. Open plastic sample bottle taking care not to
contaminate the inside of the bottle cap.
10. Add 100 ml of distilled water to the bottle.
11. Insert a sterile pipette into
the green and white extractor
tool and use it to remove 10 ml of
the distilled water from the
plastic bottle, leaving 90 ml
inside the bottle.
Page | 9
12. Discard the 10 ml of distilled water and set the
pipette and extractor aside. Don’t let the pipette touch
anything.
13. Add the Enterolert to the 90 ml of distilled water in
the plastic bottle by breaking the capsule over the
mouth of the vial. Replace the bottle cap. Shake bottle
gently back and forth for 40
seconds or until all visible
chunks are dissolved.
14. Without touching the
pipette to the baggy or the
mouth of the 100ml vial,
extract 10 ml of your sample
from the baggy/vial.
15. Add the 10 ml of sample
water to the plastic bottle
containing the Enterolert
mixture. Gently shake for 20
seconds.
16. Dispose of the plastic baggy, the plastic pipette, and
the Enterolert container. We repurpose the 100ml.
vials. Place the lids on the empty 100ml. vials and place
them in the recycling box that is under the fish tanks.
Page | 10
17. Open the Quanti-Tray by squeezing it near the top.
18. Pour the entire contents from
the plastic bottle into the
Quanti-Tray. It should fill most
of the cells.
Page | 11
19. Check to see that the green light on the Sealer is on.
20. Place the Quanti-Tray onto the red rubber mat and
feed it into the Sealer closed end first.
21. After the Quanti-Tray is sealed, confirm that the
Incubator is at 106° F or 41° C. Then place the sealed
Quanti-Tray in the Incubator.
22. Let the sample sit for 24-28 hours in the Incubator.
23. Dispose of the used gloves, bags and pipettes, rinse
and cap the 120ml bottles and put them in the plastic
lined box for repurposing. Put the green and white
extractor that fits on the pipette back in the box of
pipettes.
Quanti-Tray Filled with Complete Contents of Sample Bottle
Page | 12
THREE: ANALYZING and RECORDING DATA
1. After the incubation period is complete (24-28 hours),
remove the Quanti-Tray(s) from the Incubator.
2. Turn on the
Black Light
and scan the
Quanti-Tray
with the
Black Light.
3. Count the cells, both large and small, that light up or
“glow.”
4. Consult the chart in the binder to find the test result.
Find the number of large cells in the vertical column
down the left side, and the number of small cells in the
horizontal row across the top. At the intersection of
the appropriate column and row is the result or the
MPN (Most Probable Number). Multiply that number
by 10 (ten) to determine the actual level of
enterococcus in the original sample of ocean water in
cfu/100ml; this is the actual result of the test.
Page | 13
5. On the log sheet, record the following: (1) the number
of large cells; (2) the number of small cells; (3) the
MPN, and (4) the actual test result.
6. Communicate the results to the person who is
responsible for posting the results on the BWTF web
site. Currently this is done by taking a photo of each
completed log sheet and sending them by email to the
volunteer who is responsible for uploading the data to
the BWTF website.
7. Dispose of the Quanti-Trays. Clean up the lab area.
Turn off the Sealer and Incubator if not in use by
another BWTF volunteer.
8. If you are the last BWTF volunteer to leave and the
aquarium is open to the public, just leave. If the
aquarium is closed to the public check to determine if
there are any aquarium volunteers or staff in the
building. If staff are present, leave and make sure that
the door locks behind you. If no other volunteers or
staff are present, make sure that all exterior doors are
shut tightly and set the alarm by entering the alarm
code on the key pad and pressing ENT. The warning
beep will begin. You have 60 seconds to leave the
Page | 14
building before the alarm arms. If you have trouble and
the alarm sounds, see Section 2, Item 1.