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SOP8 – Live Organisms 10-50 μm
59
9.0. SOP 8 – Live Organisms 10 - 50 µm
Version 2.0
Date of Issuance: July 11, 2012
Author: Tim Mullady
9.1. Sampling Materials
Material
# for Uptake # for Discharge
Bottles for preserved phytoplankton
6
6
Bottles for viable phytoplankton CMFDA
6
6
Lugol’s solution (keep cold)
5 ml sample
-1
5 ml sample
-1
Formaldehyde
1ml sample
-1
1 ml sample
-1
9.2. Sampling on the Barge
Phytoplankton are collected via an integrated flow system controlled by a flowmeter and filled
over the entire course of the fill/discharge operation of each tank. The sampling rate of the
whole-water is 1.5 L minute
-1
at 300 m
3
hour
-1
ballast-flow-rate (less as flow-rates decrease). A
whole-water (not filtered) sampling-rate reference table (Table 4.2) is posted in the sample van
(see section 4.5). Water is collected in a 100L conical bottom, fiberglass, algal-culture cylinder
(referred to as whole water reservoir or collection cylinder) and aerated to keep entire water
column well mixed and in suspension. Collection cylinder is drained for 10 sec to remove any
standing water in the faucet or pipes before any collection of whole water samples are started.
Further details are found in SOP 4. Whole water reservoir cylinders are completely drained,
disconnected, thoroughly washed, and rinsed with potable water between each event.
For both fill (UT) and discharge events (DC), 3 replicate samples are taken from each tank.
Samples are collected in 500 ml, opaque, brown Nalgene bottles. Rinse empty bottles three
times with whole water from the whole water reservoir cylinder. Fill rinsed bottles the
bottleneck and place on ice packs in cooler until return to the laboratory for processing.
9.2.2
Preserved Phytoplankton & Challenge Conditions
For both fill (UT) and discharge events (DC), 3 replicate samples are taken from each tank.
Samples are collected in 500 mL opaque brown Nalgene bottles. Empty bottles are rinsed three
times with whole water from the whole water reservoir cylinder. Rinsed bottles are filled to the
shoulders of the bottle leaving enough room at the top to add preservatives. Each sample is fixed
with 5 mL of Lugol’s solution and 1 mL of formaldehyde. Samples can be set aside until
sampling is complete, and then fixed all at once. Challenge conditions for each test will be
determined by Total Phytoplankton Analysis of UT Control samples (7.5) so that an accurate
count of cell density can be determined (cells mL
-1
). This method has been in use for many years
and is much more accurate and determining the total phytoplankton community then that of
CMFDA/FDA (which determines viability). Size distribution as well as density and sinking
rates are taking into account as samples must be settled in a settling chamber (Utermöhl)
(Paxinos 2000). All preserved samples should be analyzed within a few months but can be stored
for 1 year with minimal sample degradation. Samples are kept in storage after analysis for
backup purposes and disposed of after 1 year (Modigh 2005, Stoeker 1995).