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MERC+GSI – DRAFT Test Plan
11
Test Species:
A fish, an invertebrate and a plant (algae) will be used in all ballast discharge tests.
Because the test site in Baltimore Harbor is a mesohaline aquatic environment with salinities
ranging from 5 to 15 psu, estuarine organisms will be used in these tests. The algal species will
be
Isochrysis galbana
or
Tetraselmis suecica
depending on which species performs best in
preliminary testing. The algae will be purchased from the University of Texas Algal Culture,
University of Texas, Austin, Texas. The growth media for these species will be those given in
Appendix A3 of ASTM Designation E 1218-04 “
Standard Guide for Conducting Static Toxicity
Tests with Microalgae
” (ASTM, 2006). The culture conditions will follow those given in this
guide. The fish species used in the test will be the sheepshead minnow (
Cyprinodon variegatus
)
while the invertebrate species will be the mysid (
Americamysis bahia
; formerly
Mysidopsis
bahia
). These are estuarine test species suggested for use in EPA’s Short-term Methods for
Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine
Organisms (EPA, 2002). Test organisms will be purchased from Aquatic BioSystems of Fort
Collins, Colorado. This company is our regular supplier of test organisms. They provide
excellent QA/QC, including reference toxicant testing and quality control charts for all of their
test species. Upon receipt by WREC, holding of test organisms will be conducted in accordance
to guidelines outlined in the above referenced EPA manual (2002).
Active Substance and Measurement:
The test solution will be ballast water discharged from the test tanks during each trial of
the SiCURE system. The active substance involved in this treatment is chlorine. According to
Section 5.2.8 of the IMO G9 resolution, information on Total Residual Oxidants (TRO) and
Total Residual Chlorine (TRC) should be provided as part of the application for evaluation, for
both the ballast treatment process and the ballast water discharge. The Standard Methods for the
Examination of Water and Wastewater Low-Level Amperometric Titration methods 4500-Cl D
and E will be used to measure TRO and TRC in the ballast water discharge and in the various
test dilutions. A Fischer and Porter amperometric titrator (Model 17T2000) will be used for all
measurements. By using the high-sensitivity mode, a forward titration, and a 200 ml sample,
TRO quantification limits for method 4500-Cl D are 15 µg/L TRO. With this sample size, 1 ml
phenylarsene oxide (PAO, 0.00564
N
) titrant equals 1 mg/L chlorine equivalents. For lower
levels of oxidant, method 4500-Cl E will be used. A fourfold-diluted PAO titrant (0.00141
N
)
and a strip-chart recorder for signal amplification from the Fischer and Porter amperometric
titrator (Model 17T2000) will be used to measure TRO concentrations to 5 µg/L. Samples will
be analyzed immediately upon collection onboard the
Cape Washington
to avoid loss of oxidant
due to holding. In addition to the amperometric titration method we will use a YSI Mulitmeter
(Model # 556) equipped with a probe to measure oxidation reduction potential (ORP). The
probe uses a platinum button sensor giving the instrument a range of -999 to +999 mV, an
accuracy of ± 20 mV and a Resolution of 0.1 mV.
Experimental Design and Test Conditions:
Toxicity tests will be conducted on the discharge from all test trials. As required by the
IMO G9, the discharge water will be tested with three estuarine species as described in Section
2.1. Both acute and chronic data will be generated for each test. A dilution series, using
Baltimore Harbor water, will be run for each species.