3
Statistically-validated (Miller et al., 2011), continuous, time-integrated samples were
taken for each of the following: (a) uptake water for both control and treated conditions, (b)
control and treated water upon discharge after a five-day holding time. Sample volumes and
details of the physical, chemical, and biological analyses for each sample are described below
and in MERC SOPs.
All samples collected to quantify live organisms or water quality were taken by inline
sampling of water during the entire filling or discharge of water from the tanks through sample
ports located on appropriate filling or discharge pipes. All sample ports include a valve and
sample tube with a 90
o
bend towards the direction of flow, placed in the center of the piping
system (based on the design developed and validated by the US Naval Research Laboratory, Key
West Florida, see ETV Protocols).
Water for biological examination was split for sampling the >50µm size fraction
(
nominally zooplankton), and the other fractions (10 - 50 µm size fraction, bacteria, water
quality, etc.). At the completion of each trial, test tanks were thoroughly cleaned by pressure
washer, and piping was flushed with fresh municipal water, prior to conducting the subsequent
trial. See SOPs for additional details on test operations and sampling.
Quantifying Physical Conditions:
Temperature, salinity, dissolved oxygen, chlorophyll fluorescence, and turbidity (NTU)
were measured every 15 minutes during the test trials using two identical multi-parameter probes
(
calibrated before each trial according to manufactures specification) deployed into both the
control and treated tanks. A third hand-held instrument was used to measure temperature,
salinity, and dissolved oxygen of water (and other parameters as required) during uptake or
discharge.
Arizona Test Dust, micronized humate (Micromate) and
Camellia sinensis
(
tea) (as
described in ETV Protocols and MERC SOPs) was injected inline during initial filling of control
and test tanks to increase TSS, POC and DOC levels. Initial inline samples (three replicates,
approximately 500 ml - 2 l each) of ballast water during the filling of the control and treated
tanks were collected, filtered, and analyzed for the water quality parameter total suspended solids
(
TSS). Samples for analysis of dissolved and particulate carbon and chlorophyll were collected
from the 100 L time-integrated canister. Similarly, inline samples of challenge water were
analyzed for percent UV transmittance using a UV254 P200 RealTech field spectrophotometer.
See SOPs for additional details.
When the flow rate is at or less than 100 m
3
/
hr, there is no side discharge stream at the
normal sample point for TSS and %UVT at 254nm. That is, all of the water coming from the
treated and control sample points on the pipes must be used to filter through the nets for
zooplankton or streamed into the time-integrated (whole water) water tubes for phytoplankton,
bacteria, POC, DOC and Chl. When this occurs, as with the BIO-SEA tests, the sample port for
collected challenge water for TSS and %UVT is moved to a sample port located on the vendor
pipe just prior to entering the vendor system (a few feet closer to the vendor system than the
usual sample port).
Live Organisms >50µm in size:
The sampling system consists of two sets of paired canisters, each designed to
accommodate a 35 µm (50 µm diagonal) mesh plankton net used to collect the >50 µm size
fraction. One pair handles water from the treated ballast tank and the other pair handles water