RESULTS OF TOXICITY TESTS CONDUCTED
ON EFFLUENTS, AMBIENT WATERS, AND SEDIMENTS FROM
THE LOWER CALCASIEU RIVER ESTUARY, LOUISIANA
ELISE TORELLO
MICHELE REDMOND
SCIENCE APPLICATIONS INTERNATIONAL CORPORATION
AND
GEORGE MORRISON
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
ENVIRONMENTAL RESEARCH LABORATORY, NARRAGANSETT, RI
Contribution No. 1020 of the U.S. EPA
Environmental Research Laboratory, Narragansett, RI
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ACKNOWLEDGMENTS
The authors would like to thank Robert Burgess, Pamela Comeleo,
Randy Comeleo, Wendy Greene, Kathy McKenna, Suzanne Lussier,
Steven Schimmel, Cathy Sheehan, Mark Tagliabue, Glen Thursby,
and Raymond Walsh for their hard work above and beyond the call
of duty in conducting these toxicity tests. Their assistance in
reviewing this paper is also greatly appreciated.
Mention of trade names does not constitute endorsement by the
United States Environmental Protection Agency.
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EXECUTIVE SUMMARY
The Environmental Protection Agency's Environmental Research
Laboratory at Narragansett, Rhode Island (ERL-N) participated
with EPA Region VI, U.S. Geological Survey, and the State of
Louisiana in a comprehensive study of the lower Calcasieu River.
This study, conducted during the summer of 1988, also included
two tributary bayous (Bayou Verdine and Bayou D'Inde). Staff
from ERL-N conducted a series of toxicity tests on effluents from
eight discharges, receiving waters from 24 sites, and sediments
from 35 sites.
Toxicity tests used for effluents and receiving waters were
the 2-hour sea urchin (Arbacia punctulata) fertilization test,
the 2-day macroalga (Champia parvula) reproductive test, the
mysid (Mysidopsis bahia) 7-^day survival, growth, and fecundity
test, and the larval inland silverside (Menidia beryllina) 7-day
survival and growth test. Toxicity of sediments was evaluated
with the amphipod (Ampelisca abdita) 10-day survival test. The
most sensitive test species for effluents and receiving waters
was the mysid, followed by the larval fish and sea urchin tests
in that order. Macroalga results were not used because of
unacceptable control response.
Significant toxicity was observed in six of the eight
effluents tested. Toxicity was also evident at nine receiving
water and 19 sediment sites. Toxicity, for purposes of
comparison, is arbitrarily defined here as high(ly) (toxic to two
or more of the three test species), medium-moderate (toxic to one
of the test species), or non-toxic. Sediment toxicity is
likewise classified as high (< 50% survival), medium (50 - 75%
survival), or low (> 75% survival).
Bayou Verdine was the most heavily impacted geographical
region where both of the effluent discharges tested were highly
toxic. Likewise, three of the four receiving water sites in this
bayou were highly toxic. One site, near the mouth of the bayou,
was not toxic. All five of the sediment sites in Bayou Verdine
were highly toxic.
Bayou D'Inde was somewhat less severely affected. The two
effluents tested showed low and medium toxicity. Of the eight
receiving water sites, two were moderately toxic and six were
non-toxic. Sediments in this bayou were moderately toxic at four
and highly toxic at eight of the 12 sites.
Effluents discharged into the Calcasieu River and associated
lakes (Lake Charles and Prien Lake) were highly toxic at three
and non-toxic at one of the four sites tested. Receiving waters
were non-toxic at seven and moderately toxic at four sites.
Sediments from the river and lakes were non-toxic at 12 locations
with only one site having moderate and one high toxicity.
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These toxicity test results will be incorporated with
associated sediment and water chemistry data, generated by USGS,
LADEQ, and EPA Region VI, in a final report to be written by EPA
Region VI.
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INTRODUCTION
The United States Environmental Protection Agency's
Environmental Research Laboratory in Narragansett, Rhode Island
(ERL-N) participated in a collaborative effort with four other
state and federal agencies in a study of the lower Calcasieu
River estuary, Louisiana and its tributaries, Bayou Verdine and
Bayou d'Inde. Three coastal lakes hydrologically connected to
the river (Lake Charles, Prien Lake, and Calcasieu Lake) were
also included in the study. The other participants in the
project were: the U.S.EPA Region VI, Water Management Division,
the Region VI Environmental Services Division Regional
Laboratory at Houston, the Louisiana Department of Environmental
Quality (DEQ), and the United States Geological Survey (USGS)
Water Resources Division, Louisiana District.
The study area (Figures 1 and 2) is located in an
industrialized area and is impacted by industrial stormwater
runoff and by numerous industrial and municipal point sources.
The normal course of the lower Calcasieu River is modified by
extensive channel realignment and dredging for the maintainance
of a major navigation channel. (LA DEQ and U.S. EPA, 1988)
Portions of the lower Calcasieu River and some of its
tributaries have been designated by DEQ as priority waterbodies
with known or suspected water quality problems related to toxic
pollutants. The USGS, the Louisiana Department of Health and
Hospitals (DHH), and DEQ have demonstrated the presence of
chemically contaminated sediments in localized areas and
contaminated seafood species over a wide area in the Calcasieu
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River estuary including tributaries and connected coastal lakes.
This contamination is suspected to originate from discharges
from petrochemical and organic chemical manufacturing facilities
in the Lake Charles/Calcasieu area. The DEQ has also documented
the occurrence of elevated water column concentrations of low
molecular weight halogenated volatile organics in Bayou d'Inde,
Bayou Verdine, and the near reaches of the Calcasieu River.
Efforts by DEQ, EPA Region VI, and ERL-N during 1984 and 1985
indicated the occurrence of chronic toxicity to the standard
test organisms in the ambient waters at several locations in the
Calcasieu estuary. (LA DEQ and U.S. EPA, 1988).
These data demonstrate the need for monitoring the
Calcasieu River estuary for pollutants and toxicity. The
specific objectives of ERL-N during this monitoring project
included:
1) Evaluating the utility and sensitivity of the
toxicity test methods employed;
2) Determining the occurrence and geographical extent of
any short term chronic ambient water or sediment
toxicity that may be exhibited in selected reaches of
the Calcasieu estuary during an anticipated low flow,
warm water temperature period;
3) Documenting the occurrence of any effluent toxicity
that may be exhibited by selected industrial wastewater
discharges into the Calcasieu estuary.
The results of toxicity testing will be used in conjunction with
chemical data from the estuary to determine if any relationships
between specific chemical concentrations and measured toxicity
to aquatic life exist. All data generated will be used to
provide information for the assessment and development of permit
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limits and state water quality standards.
METHODS
Sample Collection
A preliminary study to screen Calcasieu River effluents for
toxicity was conducted using the sea urchin Arbacia punctulata
and the alga Champia parvula~ Samples were collected from 14
outfalls on two days: May 23 and June 6, 1988 (Table 1). The
results of the screening studies were taken into consideration
when effluents for later sampling and testing were chosen.
Water samples from twenty-three ambient stations were
collected in addition to effluent samples from eight dischargers
(Figures 1 and 2, Tables 2-5) for definitive toxicity tests
using the full suite of marine species. Ambient water samples
were collected at mid-depth using a submersible Johnson-Keck
Trace Organics pump sampler, while all effluents were grab
sampled. Each effluent and ambient water test included three
samples collected over a one week period (Table 5). All ambient
water and effluent samples were transferred to plastic
Cubitainers and shipped on ice to ERL-N under appropriate EPA
chain-of-custody procedures. At ERL-N, samples were held at
4ฐC until they were tested. All water samples collected
over the four week test period were used within 48 hours
following collection.
Sediment samples were collected by staff of EPA Region VI,
DEQ, and USGS from thirty-five ambient sampling locations
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selected for toxicity testing (Figure 1, Tables 3 and 4). Two
grabs equally spaced across each littoral shelf plus two grabs
from the dredged channel (total of six) were composited at
stations in the Calcasieu River/Ship Channel. A minimum of
three randomly spaced grabs from 5 meter square area
(approximately) were composited at the four lake stations.
Sediment samples were collected once at each location using a
stainless steel Petite Ponar bottom sampler and were composited
and mixed in stainless steel containers. Composites were
transferred to and shipped in wide-mouthed glass jars.
Sediments for toxicity testing were shipped to ERL-N in a manner
similar to the water samples. At ERL-N, samples were held at
4ฐC until they were tested.
All of the sediments collected on a single sampling day
could not be tested concurrently due to laboratory space
constraints. Thus, it was necessary to store some sediments at
4ฐC for as long as 29 days prior to testing. A preliminary
experiment was conducted using the amphipod Ampelisca abidita to
determine the persistence of toxicity of sediments held in
refrigerated storage over an extended period of time. Sediment
samples from two stations were collected and were shipped to
ERL-N on ice. The samples were split into two portions. One
portion of each sample was immediately subjected to the standard
ten-day amphipod toxicity test. The remaining portions were
held under normal refrigerated conditions for seven weeks and
then tested. The results from the two test were compared
statistically to determine if any change in toxicity occurred in
storage.
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Another preliminary experiment was conducted to determine
if amphipod survival was affected by the salinity of a
sediment's interstitial water. Two salinities were tested in
this study: 7 parts per thousand (ppt) and 30 ppt.
Controls
Low salinity ambient and effluent samples (Tables 6 and 7)
were adjusted up to test salinity using hypersaline brine made
from Narragansett Bay water. Ambient water samples were tested
at the highest concentrations possible after salinity
adjustment. Effluent samples were also tested at the highest
concentration possible after salinity adjustment, as well as
four serial 1:1 dilutions down from the high concentration.
Salinity adjusted effluent and ambient water samples were
statistically compared to a Narragansett brine + deionized water
(brine + DI) control to assure that no toxicity resulted from
the addition of brine to samples. Station 18 was a reference
site upstream of the study area and above the US Army Corps of
Engineers saltwater barrier, and was used as the site control
for all ambient water and sediment tests. Most toxicity tests
included a performance control to demonstrate the response of
the organisms tested to water or sediment of known quality. The
performance control for water toxicity tests was filtered
Narragansett Bay water, and the sediment test performance
control was collected from an uncontaminated site in central
Long Island Sound.
The sediment tests also included a low-salinity control to
determine if the low interstitial salinity of some sediment
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samples was toxic to the amphipods. The low salinity sediment
was created by mixing performance control sediment with a
calculated amout of deionized water, allowing it settle, then
pouring off the supernatant. The supernatant water salinity was
measured to ensure that the calculated salinity was achieved.
Toxicity Tests
Toxicity tests conducted on ambient water and effluent
samples were the red macroalga (Champia parvula) reproduction
test, the sea urchin (Arbacia punctulata) fertilization test,
the mysid shrimp (Mysidopsis bahia) survival, growth, and
fecundity test, and the inland silversides (Menidia beryllina)
7-day larval survival and growth test (Tables 8 and 9) (US EPA,
1988). These tests were conducted on effluents from the
dischargers listed in Table 2 and on water from the ambient
stations indicated in Tables 3 and 4. The toxicity test
conducted on sediment samples from the Calcasieu estuary was an
acute test using the amphipod, Ampelisca abdita. This test was
conducted on sediment, samples from ambient stations 1-35. The
following method descriptions are a summary of the toxicity test
techniques employed during the project.
Champia parvula
The macroalgal (Champia parvula) reproductive test was
conducted at 30 ppt salinity and consisted of a simultaneous
2-day exposure of male and female plants to an effluent or
receiving water. The female plants were then placed in a
control medium for a 5- to 7-day recovery period during which
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Page 9
the cystocarps, evidence of sexual reproduction, developed.
Toxicity was expressed as a numerical reduction of cystocarp
production relative to the controls.
Arbacia punctulata
The sea urchin (Arbacia punctulata) fertilization test was
conducted at 30 ppt salinity. The test involved the exposure of
dilute sperm solutions to effluents or receiving waters for one
hour. Eggs were added following this exposure period and the
gametes were allowed to incubate for twenty minutes, after which
fertilization was stopped by the addition of formalin. Toxicity
was expressed as a significant reduction in percent egg
fertilization relative to the controls.
Mysidopsis bahia
The mysid (Mysidopsis bahia) short-term chronic test
involved exposing 7-day old Mysidopsis bahia juveniles to an
effluent or receiving water for 7 days. The Calcasieu River
test series was conducted at 20 ppt salinity. As a static,
renewal procedure, test water was replaced daily by the most
recently collected sample. The females matured during this
exposure period, and some produced eggs by the end of the test.
The test endpoints were growth (measured as dry weight), and
survival. Fecundity (measured as the percentage of females with
eggs), was not a consistantly acceptable endpoint and therefore
was not used in this study.
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Menidia beryllina
The inland silversides (Menidia beryllina) short-term
chronic test was conducted at 20 ppt salinity and consisted of
the exposure of 7-9 day old larvae to effluents or receiving
waters for 7 days. As in the mysid procedure, exposure water
was replaced daily with the most recently collected sample. The
test endpoints for this method were growth (measured as dry
weight per fish) and survival.
Ampelisca abdita
The acute tests with this benthic amphipod consisted of
ten-day exposures of juvenile amphipods to samples from each of
the 35 Calcasieu River ambient stations. The samples were
divided into three groups for testing, since space constraints
precluded the simultaneous testing of 35 samples. Sediments,
after they were press sieved (2 mm) to remove large debris and
potential predator organisms, were homogenized and added to the
exposure chambers. Each exposure chamber contained 30 juvenile
amphipods, and approximately 200 ml of sediment and 600 ml of
seawater. There were three replicates per treatment. The water
column in this flow-through test was filtered and aerated
Narragansett Bay water. The animals were not fed during the
exposure period. Sediments were screened and sorted at the
conclusion of the test to determine the number of survivors in
each treatment replicate. The test endpoint was a statistical
reduction in survival relative to the controls. See Appendix a
for a more detailed description of the Ampelisca abdita test
methodology.
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Data Analysis
Data from all of the toxicity tests were subjected to a
one-way analysis of variance (ANOVA). Series of "T" Tests were
performed on sediment and ambient water toxicity test data,
while effluent tests were analyzed using Dunnett's Test
(Dunnett, 1955). Arcsine square root transformations were
performed on the sea urchin and amphipod data, the silversides
and mysid survival data, and on the data relating to mysid
percent females with eggs before statistical tests were
conducted. Abbott's formula was used to correct for control
amphipod mortality before arcsine transformation and analysis by
Fisher's protected least significant difference test in the
preliminary sediment storage experiment. No transformations
were performed on the mysid or silversides weight data prior to
analysis.
Effluent test results were expressed as the lowest observed
effect concentration (LOEC) and no observed effect concentration
(NOEC) for each effluent, while ambient water and sediment test
results were expressed as significant toxic effects compared to
the controls.
RESULTS
Champia parvula
The first screening test of effluents was not acceptable
due to poor control response (<10 cystocarps per plant). The
second screen included two concentrations per effluent: 1ฐ? and
10%. Effects were evident in 10% effluent from Citgo outfalls
001 and 003, W.R. Grace outfall 001, Firestone outfall 001,
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Vista outfall 001, Westlake Polymers outfall 007, and PPG
outfall 001 (Table 1). No toxicity was evident at 1% in any of
these effluents. Effects were observed at 1% and 10% in
effluents from Oxy outfall 002E, Olin outfall 010, and Conoco
outfall 001. Effluent from Citgo 003, W.R. Grace 001, Vista
001, Olin 010, and Conoco 001 was acutely toxic to Champia (<_1
cystocarp per plant) at 10%. No effects were observed in
effluent from Himont outfall 001, PPG outfall 004, or Westlake
Polymers outfall 001.
Due to difficulties in maintaining the quality of
laboratory Champia cultures during the study period, none of the
definitive test controls produced enough cystocarps to maintain
test acceptability.
Arbacia punctulata
Four of the twelve effluent samples tested during the first
screening were toxic to Arbacia (Table 1). No observed effect
concentrations (NOECs) for effluents from W.R. Grace outfall
001, PPG outfall 001, Olin outfall 028, and Vista 001 were
17.5%, 35.0%, 17.5%, and 8.8%, respectively. Concentrations
which would reduce fertilization by 50% relative to the controls
(EC50s) were calculable for effluents from Olin 028 and Vista
001; these values were 47.36% and 23.91% respectively.
Three of the thirteen effluents tested during the second
screening were toxic to Arbacia (Table 1). Effluent from w.r.
Grace 001 yielded a NOEC value of <35.0%, and effluents from PPG
001 and Firestone 001 had NOECs of 35.0%. EC50s for these
effluents were 48.91%, 51.92%, and 64.72%, respectively. NOECs
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for effluents from W.R. Grace 001 and PPG 001 were similar in
both screenings.
Most of the effluents tested definitively were toxic to
some degree to the sea urchin; however, a dose effect was
evident on only one occasion (Tables 10 and 11). Effluent from
Olin outfall 001 collected 7/11 yielded an EC50 of 53.6 percent
effluent. No significant effects were observed at any time in
effluent from PPG outfall 001, Olin outfall 028, or Citgo
outfall 001.
About a third of the ambient water samples tested during
the first week of the study (Stations 3-18) caused statistically
significant reductions in fertilization; however, the toxicity
observed was slight (<10% reduction from the controls) (Table
13). Moderate toxicity was evident on one occasion, when
fertilization in sample from Station 4 collected on 6/20 was
reduced to 75.9%. Several samples collected during the second
week of the study (Stations 18-34) caused significant effects,
but again, the toxicity was slight except on one occasion (Table
15). Toxicity was observed in sample collected from Station 19
on 6/29, when fertilization was reduced to 53.6%.
Mysidopsis bahia
Six of the eight effluents tested were toxic to the mysids
(Tables 16-19). Effluents from Vista Chemical 001 (Table 16)
and Conoco, Inc. 001 (Table 18) were the most toxic tested.
Toxic effects were observed in these effluents at 5%, the lowest
effluent concentration tested. Vista and Conoco effluents
caused 0% survival at 42% and 20%, respectively. Effluents from
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Olin outfalls 001 and 010 (Table 17) and Citgo outfall 003
(Table 19) were also quite toxic, with toxic effects evident at
10%, 11%, and 10.5%, respectively. These effluents caused 0%
survival at 40%, 22%, and 84%, respectively. Fecundity and
final weights were reduced in Citgo outfall 001 (Table 19)
effluent at 20% and 40%, respectively, but there was no
significant effect on survival at the highest concentration
tested. No toxic effects were observed in effluent from PPG
Industries 001 (Table 16) or Olin outfall 028 (Table 18).
Toxic effects were observed in nine of the 23 ambient
waters tested (Tables 20 and 21). Samples from Stations 13, 15,
19, 20, 21, and 26 all caused toxic effects when compared to the
site control (Station 18). Water from Station 20 caused 0%
survival. Reduced survival was observed in samples from
Stations 3, 5, and 18 when compared to the brine + DI control.
Menidia beryllina
Five of the eight effluents tested were toxic to Menidia
(Tables 22-25). Effluent from Vista Chemical 001 (Table 22)
caused reduced growth at 21% and reduced survival at 83%.
Effluent from Olin Corporation outfall 001 (Table 23) affected
survival of larvae at 40.0% and caused 0% survival at 81.0%.
Effluent from Olin outfall 010 (Table 23) caused 0% survival at
22.0%. Conoco, Inc. 001 (Table 24) effluent caused reduced
growth at 5.0%, the lowest concentration tested, and caused 0%
survival at 41.0%. Citgo 003 (Table 25) effluent was quite
toxic, causing reduced growth at 10.5% effluent and 0% survival
at 84.0% effluent. Effluents from PPG Industries (Table 22),
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Olin Corporation outfall 028 (Table 24), and Citgo Petroleum
outfall 001 (Table 25) were not toxic to Kenidia beryllina.
Some toxicity was evident in three of the twenty-three
ambient waters tested using the silverside (Tables 26 and 27).
The sample from Station 20 was highly toxic, with a mean
survival of 2.2%. Growth was significantly reduced in samples
from Stations 19 and 21, and survival was significantly reduced
in the sample from Station 20.
Ampelisca abdita
No significant reduction of toxicity due to storage was
evident (Table 28). No significant difference in amphipod
survival was observed between sediments with low and high
interstitial water salinities (Table 28).
Most of the sediments tested during the definitive study
were toxic to Ampelisca (Tables 29-31). The only samples tested
which did not reduce survival compared to Station 18 were from
Stations 8, 13-17, 24, 26-28, and 31-35. Samples from Stations
3A-C, 4, 9, 12, 29, and 30 were moderately toxic, with mean
percent survival values between 30.0% and 77.8%. Samples from
Stations 2, 5-7, 19, and 25 were highly toxic, with mean percent
survival values below 10%. Samples from Stations 1, 10, 11, and
20-23 caused 0% survival.
Replicates to be tested separately were taken at Stations 3
and 21. Three replicates were tested from Station 3, and all
were moderately toxic. Replicates B and C were significantly
different from each other. The two replicates tested from
Station 21 both caused 0% survival.
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CONCLUSIONS
Three of the four water toxicity test methods yielded
useful results when all of the data were examined. The
Mysidopsis bahia and Menidia beryllina tests proved to be
especially sensitive to the effluents and ambient waters tested,
and the results of the two test procedures correlated very
closely (Tables 32 and 33). The first two mysid tests, however,
did not meet the criterion for acceptability for control dry
weights. When young mysids are held in tanks that are too
densely populated during the 7-days prior to a test, they do not
grow as quickly as they do at optimum population density. We
believe that the failure to meet the acceptability criterion was
caused by these overcrowded conditions, and that the mysids were
otherwise healthy and the test results were valid.
One disturbing result of the first week of mysid ambient
water testing was that the site control (Station 18) appeared to
be somewhat toxic. This suggests a need to evaluate the use of
this station as a reference in future testing.
One of the most toxic effluents tested was from Olin
outfall 010. This effluent was quite distinctive in that it
possessed a strong green color (believed to be due to the
presence of Chlorella) and was highly odorous. 100% mortality
was evident in both the Menidia and mysid tests at 81% and 40%,
respectively, within 48 hours of test initiation. Several
fractionations were performed on this effluent using Menidia
tests on the fractions, but the results were inconclusive.
The amphipod test proved to be very sensitive to
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contaminated Calcasieu River sediments. In fact, 60% of the
sediments tested were toxic to Ampelisca. Sediment toxicity was
accompanied by water column toxicity at five stations (Table
33). The significant difference in survival between two
replicates at Station 3 indicates possible patchy distribution
of toxicants in the sediment; however, it is difficult to draw
firm conclusions on one sample.
Relationships between effluent toxicity and sediment and/or
ambient water toxicity are apparent at most of the stations
where toxicity was evident. All three sediment samples from
stations associated with the PPG outfall were toxic, although
the effluent itself was not found to be toxic to the water
column species tested at that time. Sediment from Station 1 (at
the PPG 001 outfall) was acutely toxic to Ampelisca, but
toxicity decreased with distance downstream from the PPG
outfall. Stations 5 through 9 are downstream of Citgo outfall
001, effluent from which was found to be toxic to the mysids.
Stations 19 and 20 are upstream and downstream, respectively, of
Vista outfall 001, and sediment samples from these stations were
toxic to Ampelisca. Effluents from Vista and Conoco 001 were
toxic to both Menidia and the mysids. Stations 21 through 23
are downstream of both Vista 001 and Conoco outfall 001, and
sediment from these stations was toxic to Ampelisca. Likewise,
water from Station 21 was toxic to the mysids and Menidia.
Sediment from Station 25 was toxic to Ampelisca. Station 25 is
immediately downstream of Olin outfall 010, effluent from which
was toxic to the mysids and Menidia. Sediment from Station 30,
downstream of Citgo outfall 003, was toxic to the amphipods.
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This outfall discharged an effluent which was toxic to the
mysids and Menidia.
The most severely impacted section of the Calcasieu River
estuary based on toxicity test results was Bayou Verdine.
Ambient water from Stations 19-21 in this Bayou was toxic to
both the mysids and Menidia, and sediment from Stations 19-23
caused amphipod survival to be reduced to 1.1% or less. Coon
Island splits Bayou Verdine into two channels after Station 23.
Station 24 is located in the west channel, while Station 25 is
located in the east channel. Sediment from Station 24 was not
toxic to the amphipods, but sediment from Station 25 reduced
amphipod survival to 8.9%. This indicates a possible tendency
for more of the toxicants from upstream to follow the eastern
channel around Coon Island than the western channel.
The most sensitive species used to test Calcasieu River
ambient waters and effluents was Mysidopsis bahia. Sensitivity
of the mysid test was equal to or greater than the Menidia and
Arbacia tests for all of the effluents tested. Menidia were
sensitive to three of the ambient waters tested, while mysids
were sensitive to these three plus an additional six stations.
Five of these nine stations (Stations 5, 19-21, and 26) were
downstream of effluent discharges which proved to be toxic to
Menidia and/or the mysids.
Survival appeared to be a more sensitive endpoint than
growth in the mysid test when ambient water toxicity was
detected. Growth was the more sensitive endpoint in the flenidia
test. The mysid endpoints of survival and growth were equally
sensitive in the effluent tests, whereas the Menidia growth
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endpoint was again more sensitive than survival.
In recommending species for further testing, the
sensitivity of the test methods employed is an important
consideration. The Arbacia punculata test tended not to be
sensitive to the ambient waters tested and effluent toxicity
results were for the most part inconclusive. The Ampelisca,
mysid, and Menidia tests, however, yielded very good results and
we strongly recommend their use in further testing.
We believe that the toxicity testing portion of this
project was highly successful in demonstrating effluent, ambient
water, and sediment toxicity and their interrelationships. The
results of this testing will also be very useful in comparing
chronic toxicity with chemistry data from other portions of this
project.
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REFERENCES
Dunnett, C.W. 1955. A multiple comparisons procedure for
comparing several treatments with a control.
JASA 50:1096-1101.
LA DEQ, U.S. EPA. 1988. Sampling and Analysis Project
Plan for a Biotoxicity and Chemical Contaminants
Characterization of the Lover Calcasieu River
Estuary, Louisiana.
Scott, K.J., and M.S. Redmond. in press. The effects of
a contaminated dredged material on laboratory
populations of the tubicolous amphipod,
Ampelisca abdita. Aquatic toxicology and
hazard assessment: Twelfth Symposium.
American Society for Testing and Materials,
Philadelphia, PA.
U.S. EPA. 1988. Short-Term Methods for Estimating the
Chronic Toxicity of Effluents and Receiving Waters
to Marine and Estuarine Organisms. Weber, C.I.,
et al (eds). EPA Office of Research and
Development EPA-600/4-87/028 (May 1988).
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Salt Water Barrier
Cooney
Island
Bayou Verdine
22 25
Lake Charles
Coon Island
24
Bayou d'lnde
Contraband Bayou
29
Prien Lake
30
Moss Lake
Bayou Guy
32
Olson Bayou
Black Bayou
Choupique Island
Choupique Bayou
34
Ship Channel
Calcasieu Lake
Figure 1. Ambient sampling locations for the Calcasieu River study.
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Salt Water Barrier
CONOCC 001
VISTA 001
OLIN 028 I
\OLIN 001 }
Lake
Charles
Bayou Verdine
OLIN 010 '
PPG 001
Clooney Island
Contraband
Bayou
Coon Island
Bayou d'lnde
Prien
V Lake
CITGO 001
CITGO 003 J
Figure 2. Effluent discharges sampled for the Calcasieu River study.
-------
Table 1. Results of Calcasieu River effluent screening evaluations
using the alga Champia parvula and the sea urchin Arbacia
punctulata. Results are presented as no observed effect
concentrations (NOECs) for each effluent on each collection day.
Estimated concentrations at which fertilization would be reduced by
50% relative to the controls (EC50s) are in parentheses. EC50s were
not calculable for most of the effluents tested.
Effluent
C. parvula
A. punctulata
6/6/88
5/23/88
6/6/88
Citgo 001
1.0%
>70.0%
>70.0%
Citgo 003
1.0%
>70.0%
>70.0%
W.R. Grace 001
1.0%
17.5%
<35.0% (48.
91)
Himont 001
>10.0%
>70.0%
>70.0%
Oxy 002E
<1.0%
>70.0%
>70.0%
PPG 001
1.0%
35.0%
35.0% (51.
92)
PPG 004
>10.0%
>70.0%
>70.0%
Olin 010
<1.0%
>70.0%
>70.0%
Olin 028
17
.5% (47.36%)
Westlake 001
>10.0%
>70.0%
>70.0%
Westlake 007
1.0%
>70.0%
Vista 001
1.0%
8
.8% (23.91%)
>70.0%
Conoco 001
<1.0%
>70.0%
Firestone 001
1.0%
>70.0%
35.0% (64.
72)
-------
Table 2. Descriptions of the permitted industrial
sampled during the Calcasieu River, LA study.
wastewater discharges
DISCHARGE
OUTFALL
NUMBER
NPDES
NUMBER
RECEIVING
WATERBODY
PPG Industries, Inc.
Vista Chemicals
Olin Corporation
Olin Corporation
Olin Corporation
Citgo Petroleum Corp,
Citgo Petroleum Corp,
Conoco, Inc.
001
001
001
010
028
001
003
001
LA0000761
LA0003336
LA0005347
LA0005347
LA0005347
LA0005941
LAO005941
LA0003026
PPG Canal/
Bayou d'Inde
Bayou Verdine
Calcasieu River
Clooney Island Loop
Calcasieu River
Coon Island Loop
Calcasieu River
Clooney Island Loop
Bayou d'Inde
Calcasieu River
Bayou Verdine
-------
Table 3. Descriptions of the stations sampled for toxicity testing during
the Calcasieu River, LA study. Sediment samples were collected at all of
these stations. Stations where ambient water samples were also collected
are marked (*).
STATION DESCRIPTION 3
1 * PPG Canal immediately downstream of Mobil Bridge 2 (PPG 001)
2 PPG Canal immediately downstream of Mobil Bridge 3, 1/4 mile
upstream from Bayou d'Inde
3 * PPG Canal at mouth
4 * Bayou d'Inde 200 yards downstream Little Bayou d'Inde
5 * Bayou d'Inde 200 yards downstream of CITGO 001
6 * Bayou d'Inde immediately downstream of Firestone 001
7 Bayou d'Inde 150 yards upstream of LA Hwy. 108
8 * Bayou d'Inde immediately downstream of LA Hwy. 108
9 * Bayou d'Inde 1/4 mile upstream of PPG Canal
10 Bayou d'Inde 1/4 mile downstream of PPG Canal
11 * Bayou d'Inde 1/2 mile downstream of PPG Canal, 1/4 mile
upstream of Calcasieu Ship Channel
12 * Bayou d'Inde at mouth
13 * Calcasieu River Ship Channel adjacent to mouth of Bayou d'Inde
14 Prien Lake at mouth of cut from Calcasieu River Ship Channel
15 * Prien Lake in littoral area along western shoreline midway
between Ship Channel "cut" and "outlet"
16 Prien Lake Outlet
17 * Lake Charles at Rangia reef directly east of Buoy 130 and
south of Lake Charles public beach
18 * Calcasieu River at U.S. Hwy. 171 near Moss Bluff
a From Calcasieu River study project plan.
-------
Table 4. Stations sampled from June 27 - July 1, 1988 for toxicity testing
during the Calcasieu River, LA study. Sediment samples were collected at all
of these stations. Stations where ambient water samples were also collected
are marked (*). Station 18 was sampled again as a site control.
STATION DESCRIPTION 9
19 * Bayou Verdine immed. upstream of Vaughn Rd. Bridge (Westlake)
20 * Bayou Verdine approx. 1/2 mi. downstream of Truesdale Rd.
(downstream of Vista 001 and upstream of Conoco 001)
21 * Bayou Verdine at Interstate 10
22 Bayou Verdine at road approx. 1/2 mi. upstream from Coon Is.
Loop barge slip
23 * Bayou Verdine at mouth (barge slip)
24 * Calcasieu River Coon Is. Loop midstream adjacent to PPG
South Terminal Dock (west side of loop)
25 * Calc. R. midstream and approx. 200 yds. SSE of Olin 010 (east
side of loop)
26 * Calc. R. Clooney Is. Loop midstream adjacent to "Mike Hooks"
dock (east side of loop)
27 * Calc. R. Clooney Is. Loop midstream adjacent to SW corner of
Clooney Is. (west side of loop)
28 Calc. R. at Buoy 112 between Port of Lake Charles and Prien Lake
29 Calc. R. Ship Channel at Buoy 108, approx. 1.3 mi. SW and down-
stream of Bayou d'Inde
30 * Calc. R. at Bouy 106, approx. 1/4 mi. upstream of Vincent Landing
31 * Calc. R. immed. due E of "SOHIO" stack, approx. 1/4 mi. N of
Bouy 104
32 Calc. R. at Buoy 96, adj. to SE outlet from Moss Lake
33 Calc. R. at Bouy 90, approx. 1/2 mi. S of Gulf Intracoastal
Waterway (GIWW)
34 * Calcasieu Lake, West Pass immed. NE of Cutoff Point (northern
end of Calc. Lake)
35 Calc. Lake, mid-lake approx. two mi. W of Commissary Point
a From Calcasieu River study project plan.
-------
Table 5. Schedule of sampling dates during the Calcasieu River, LA study.
COLLECTION DATES
6/20a,6/22,6/24 6/27a,6/29,7/01 7/06,7/08,7/11 7/11,7/13,7/15
PPG Indus
t. 001
Vista Chem
RW
2
RW
18
b
RW
3
b
RW
19
b
RW
4
b
RW
20
b
RW
5
b
RW
21
b
RW
6
b
RW
22
RW
7
RW
23
b
RW
8
b
RW
24
b
RW
9
b
RW
25
b
RW
10
RW
26
b
RW
11
b
RW
27
b
RW
12
b
RW
28
RW
13
b
RW
29
RW
14
RW
30
b
RW
15
b
RW
31
b
RW
16
RW
32
RW
17
b
RW
33
RW
18
b
RW
34
b
RW
35
Olin Corp. 001 Conoco, Inc. 001
Olin Corp. 010 Citgo Petro. 001
Olin Corp. 028 Citgo Petro. 003
a
b
Indicates days on which sediment samples were collected,
samples were collected at all stations.
Indicates ambient water sampling sites.
Sediment
-------
Table 6. Estimated salinities (parts per thousand) of
effluents collected during the Calcasieu River, LA study based
on the refraction of light.
COLLECTION
EFFLUENT
1
2
3
MEAN
PPG Industries
16
16
18
16.7
Vista Chemicals
4
4
4
4.0
Olin Corp. 001
0
0
2
0.7
Olin Corp. 010
16
8
8
10.7
Olin Corp. 028
0
0
0
0.0
Conoco, Inc.
2
2
2
2.0
Citgo Petro. 001
0
0
0
0.0
Citgo Petro. 003
5
5
6
5.3
-------
Table 7. Salinities (ppt) of ambient waters
collected for toxicity testing during the
Calcasieu River, LA study.
COLLECTION
STATION 12 3 MEAN
3
16
16
16
16.0
4
11
8
9.0
5
11
8
12
10.3
6
12
11
11
11.3
8
12
12
12
12.0
9
13
15
15
14.3
11
12
15
16
14 . 3
12
12
16
14
14.0
13
22
22
22
22.0
15
14
12
14
13.3
17
8
8
8
8.0
18
0
0
0
0.0
18
1
0
0
0.3
19
1
4
2
2.3
20
1
4
4
3.0
21
3
6
5
4.7
23
11
12
10
11.0
24
16
16
15
15.7
25
10
8
8
8.7
26
21
16
15
17.3
27
21
16
16
17.7
30
18
18
17
17.7
31
20
18
17
18.3
34
20
18
18
18.7
-------
Table 8.
Calcasieu
A summary
River, LA
of the toxicity test methods employed in the
study .
SPECIES
TYPE OF
TEST
LENGTH OF
EXPOSURE
TEST
ENDPOINTS
Champia parvula
Effluent/
Receiving water
48 hours
Reproduction
Arbacia punctulata
Effluent/
Receiving water
60 minutes
Fertilization
Mysidopsis bahia
Effluent/
Receiving water
7 days
Survival
Growth
Fecundi ty
Nenidia beryllina
Effluent/
Receiving water
7 days
Survival
Growth
Ampelisca abdita
Sediment
10 days
Survival
a See U.S. EPA (1988).
-------
Table 9. Schedule of Champia parvula and Arbacia punctulata tests
performed on Calcasieu River ambient water and effluent samples.
Collection
Date C_^ parvula A. punctulata
6/20/88
XX
XX
6/22/88
XX
XX
6/24/88
XX
6/27/88
XX
XX
6/29/88
XX
XX
7/01/88
7/06/88
XX
XX
7/08/88
XX
7/11/88
XX
XX
7/13/88
XX
XX
7/15/88
-------
Table 10. Results of Calcasieu River effluent evaluations conducted
during June, 1988 using the sea urchin, Arbacia punctulata. Results are
presented as percent of eggs fertilized on each collection day. Controls
included in the test were a Narragansett Bay natural seawater control
(NSW) and a Narrgansett brine + deionized water control (BR + Dl).
EFFECT, PERCENT FERTILIZED
EFFLUENT,
(%)
6/20
6/22
6/24
6/27
6/29
CONTROL
(NSW)
97.7+0.6
98.0+1.7
98.3+0.6
92.0+2.6
91.3+1.2
CONTROL
(BR + DI)
96.3+2.3
97.3+1.5
99.0+0.0
81.3+2.1
90.0+6.1
PPG IND. a
70.0%
97.7+0.6
.
83.3%
97.3+0.6
85.4%
94.7+2.9
VISTA CHEM.
4.6%
9.1%
18.2%
36.5%
72.9%
76.3+11.7
29.5+6.4 b
23.6+12.2 b
24.1+22.8 b
15.4+2.7 b
88.7+2.3
82.5+0.9
61.0+7.9
57.7+1.2 i
54.3+5.0 1
a Lower concentrations not included since no effects were observed,
b Significantly lower than the BR + DI control.
-------
Table 11. Results of Calcasieu River effluent evaluations conducted
during July 1988 using the sea urchin, Arbacia punctulata. Results
presented as percent of eggs fertilized on each collection day.
EFFECT, PERCENT FERTILIZED
EFFLUENT,
(%) 7/06 7/11 7/13
NSW
73.1
+
7.1
95.0
+
2.0
93.7
+
1.5
BR + DI
85.0
+
3.0
94.7
+
4.5
95.0
+
2.0
OLIN 001
4.4%
64.0
+
8.9 a
8.8%
63.3
+
6.7 a
-
17.5%
57.3
+
19.7 a
-
35.0%
60.3
+
15.0 a
--
70.0%
61.3
+
2.5 a
17.9%
94.0
+
1.7
b
_ ,
m m. m
35.7%
82.8
+
6.6
a
--
71.4%
-ฆ
21.0
+
4.6
a
--
OLIN 010
41.7%
90.0
+
3.0
83.3%
92.3
+
1.5 a
76.1%
-ฆ
94.0
+
1.0
OLIN 028
4.4%
96.0
+
1.0
8.8%
90.3
+
1.5
-
17.5%
86.8
+
8.2
-
35.0%
81.9
+
5.4
-
70.0%
68.0
+
26.3
95.7
+
0.6
CITGO 001
70.0%
98.3
+
0.6
97.7
+
0.6
CITGO 003
18.6%
92.0
+
1.0
-
37.0%
80.3
+
4.9
a
96.7
+
0.6
74.5%
60.0
+
2.6
a
77.0
+
2.6
CONOCO, INC.
8.9%
--
-
93.7
+
0.6
17.9%
-
97.0
+
2.6
84.0
+
7.2
35.7%
--
82.7
+
1.5
a
59.7
+
4.7
71.4%
~
81.0
+
1.7
a
51.9
+
7.9
a Significantly lower than the BR + DI control,
b EC50 - 53.6 + 2.2
-------
Table 12. Concentrations of receiving water samples from Stations 3-18
ฆrequiring salinity adjustment in tests using the sea urchin Arbacia
punctulata on each collection day.
CONCENTRATION (%)
STATION 6/20 6/22 6/24 MEAN
3
83.3
83.3
83.3
83.3
+ 0.0
4
79.5
76.1
76.1
77.2
+ 2.0
5
79.5
76.1
79.5
78.4
+ 2.0
6
79.5
79.5
79.5
79.5
+ 0.0
8
79.5
79.5
79.5
79.5
+ 0.0
9
81.4
83.3
85.4
83.4
+ 2.0
11
79.5
83.3
83.3
82.0
+ 2.2
12
79.5
83.3
81.4
81.4
+ 1.9
13
89.7
89.7
89.7
89.7
+ 0.0
15
81.4
79.5
81.4
80.8
ฑ 1-1
17
76.1
76.1
76.1
76.1
+ 0.0
18
70.0
70.0
70.0
70.0
+ 0.0
(SITE CONTROL)
-------
Table 13. Results of Calcasieu River ambient water evaluations using the
sea urchin, Arbacia punctulata. Results are presented as percent of eggs
fertilized on each collection day. Controls included in the test were a
Narragansett Bay control (NSW), a Narrgansett brine + deionized water
control (BR + DI), and a site control (Station 18).
EFFECT, PERCENT FERTILIZED
STATION 6/20 6/22 6/24 MEAN
CONTROL 97.7+0.6 98.0+1.7 98.3+0.6 98.0+0.3
(NSW)
CONTROL 96.3+2.3 97.3+1.5 99.0+0.0 97.5+1.4
(BR + DI)
3
99.0+1.0
96.7+2.3
98.3+0.6
98.0+1.2
4
75.9+5.2
a, b, c
97.7+1.5
98.3+0.6
91.6+12.8
5
97.3+2.3
97.0+2.0
97.7+0.6
b
97.3+0.4
6
97.7+0.6
97.7+0.6
97.7+2.1
97.7+0.0
8
98.7+0.6
97.3+1.5
95.7+2.5
b
97.2 + 1.5
9
93.3+2.1
a, c
96.0+3.0
95.0+1.0
a, b
94.8+1.4
11
98.0+1.7
90.6+4.9 a
89.3+1.2
a, b, c
92.6+4.7
12
97.7+0.6
94.7+3.1
93.0+1.0
a, b
95.1+2.4
13
95.1+3.3
88.7+6.1 a,b
93.3+3.2
a,b
92.4+3.3
15
96.7+1.5
95.7+1.5
90.7+0.6
a ,b
94.4+3.2
17
95.3+0.6
a, c
96.7+1.2
93.0+2.0
a, b
95.0+1.9
18
98.0+0.0
96.7+3.0
94.7+2.3
a,b
96.5+1.7
(SITE CONTROL)
a Significantly lower than the NSW control,
b Significantly lower than the BR + DI control,
c Significantly lower than the Site control.
-------
Table 14. Concentrations of receiving water samples from Stations 18-
requiring salinity adjustment in tests using the sea urchin Arbacia
punctulata on each collection day.
CONCENTRATION, (%)
STATION 06/27/88 06/29/88 MEAN
18 71.4 70.0 70.7 + 1.0
(SITE CONTROL)
19 71.4 72.9 72.2 + 1.1
20 71.4 72.9 72.2 + 1.1
21 72.9 74.5 73.7 + 1.1
23 79.5 79.5 79.5 + 0.0
24 83.3 83.3 83.3 + 0.0
25 77.8 76.1 77.0 + 1.2
26 89.7 83.3 86.5 + 4.5
27 89.7 83.3 86.5 + 4.5
30 85.4 85.4 85.4 +0.0
31 87.5 85.4 86.5 + 1.5
34 87.5 85.4 86.5 + 1.5
-------
Table 15. Results of Calcasieu River ambient water evaluations using the
sea urchin, Arbacia punctulata. Results are presented as percent of eggs
fertilized on each collection day. Controls included in the test were a
Narragansett Bay control (NSW), and a Narrgansett brine + deionized water
control (BRINE + DI) .
EFFECT, PERCENT FERTILIZED
STATION 06/27/88 06/29/88 MEAN
CONTROL
(NSW)
92.0
+
2.6
91.3
+
1.2
91.7
+
0.5
CONTROL
(BRINE + DI)
81.3
+
2.1
90.0
+
6.1
85.7
+
6.2
18
(SITE CONTROL)
91.7
+
1.2
96.0
+
1.0
93.9
+
3.0
19
89.3
+
1.5
53.6
+
1.4
a, b, c
71.5
+
25.
20
90.5
+
0.9
93.3
+
1.5
91.9
+
2.0
21
92.3
+
0.6
95.7
+
3.2
94.0
+
2.4
23
87.6
+
2.2
b
93.3
+
2.3
90.5
+
4.0
24
86.7
+
5.9
94.7
+
3.5
90.7
+
5.7
25
87.7
+
1.5
b
95.0
+
1.0
91.4
+
5.2
26
84 .7
+
1.5
a, b
93.7
+
2.3
89.2
+
6.4
27
88.3
+
4.0
90.7
+
1.5
b
89.5
+
1.7
30
87.0
+
1.7
b
89.3
+
2.1
b
88.2
+
1.6
31
84.6
+
3.2
a, b
92.3
+
2.1
b
88.5
+
5.4
34
95.6
+
1.4
91.7
+
2.1
b
93.7
+
2.8
a Significantly lower than the NSW control,
b Significantly lower than the Site control,
c Significantly lower than the BR + DI control.
-------
Table 16. Effects on the mysid shrimp (Mysidopsis bahia) during the 7-day laboratory exposure to
effluents from PPG Industrial and Vista Chemical during the Calcasieu River study. Effects measured
are growth (weight) and survival. Fecundity was not an acceptable endpoint of either test.
PINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY UT/ TEMPERATURE SALINITY D.O.
(%) (Z) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
PPG IND. 001
BRINE + DI
87.5
0.163
+
0.027
26.1
+
0.3
21.0
+
0.8
5.8
+
0.5
6.0
90.6
0.171
+.
0.036
26.0
+
0.1
21.0
+
0.9
5.7
4-
0.4
12.0
90.6
0.177
+
0.020
26.1
~
0.2
21.0
+
0.9
5.8
+
0.4
24.0
100.0
0.159
+
0.024
26.0
+
0.3
21.0
+
0.8
5.8
+
0.4
48.0
96.9
0.153
+ฆ
0.037
25.9
+
0.3
21.0
+
1.0
5.8
+
0.4
96.0
84.4
0.166
+
0.022
25.9
+
0.1
20.0
+
0.7
5.8
+
0.3
VISTA CHEM. 001
BRINE + DI
87.5
0.155
+
0.031
25.9
+
0.3
21.0
-------
Table 17. Effects on the mysid shrimp (Mysidopsis bahia) during the 7-day laboratory exposure to
effluents from Olin Corp. outfalls 001 and 010 during the Calcasieu River study. Effects measured
are growth (weight) and survival. Fecundity was not an acceptable endpoint for either test.
FINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY UT/ TEMPERATURE SALINITY D.O.
(X) (2) INDIVIDUAL
-------
Table 18. Effects on the mysid shrimp (Hysidopsis bahia) during the 7-day laboratory exposure to
effluents from Olin Corp. outfall 028 and Conoco, Inc. during the Calcasieu River study. Effects
measured are growth (veight) and survival. Fecundity was not an acceptable endpoint for either test.
PINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY VT/ TEMPERATURE SALINITY D.O.
(X) (Z) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
OLIN 028
BRINE + DI
100.0
0.200
+
0.020
26.0
0.7
22.0
+
0.6
5.3
+
0.7
5.0
97.5
0.213
+
0.026
26.0
+
0.4
21.0
+
0.9
5.5
+
0.6
10.0
97.5
0.222
+
0.018
26.0
+
0.7
21.0
0.9
5.5
+
0.5
20.0
97.5
0.217
+
0.027
25.8
0.6
21.0
+
1.0
5.4
0.7
40.0
97.5
0.229
0.029
26.1
0.5
21.0
0.7
5.4
+
0.5
80.0
95.0
0.236
+
0.015
25.9
+
0.5
21.0
+
1.1
5.4
0.7
CONOCO, INC. 001
BRINE ~ DI
96.9
0.233
+
0.035
26.7
0.1
21.0
+
1.4
5.7
~
0.9
5.0
87.5
0.173
+
0.031 a
26.5
+
0.2
22.0
+
0.7
5.3
+
0.5
10.0 b
25.0 a
0.144
+ฆ
0.045
26.5
+
0.2
21.0
~
0.9
5.4
+
0.5
20.0
0.0 a
26.4
0.2
21.0
A
0.8
5.4
0.4
+
T
+
41.0
0.0 a
26.4
A.
0.1
21.0
0.9
5.5
0.2
"T
T
+
82.0
0.0 a
26.4
+
0.2
21.0
1.0
5.3
0.4
+
+
a Significantly lower than the brine + DI control.
b Veight data were not analyzed at this and higher concentrations since survival effects were observed.
-------
Table 19. Effects on the mysid shrimp (Mysidopsis bahia) during the 7-day laboratory exposure to
effluents from Citgo Petroleum outfalls 001 and 003 during the Calcasieu River study. Effects
measured are growth (weight) and survival.
FINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY UT/ TEMPERATURE SALINITY D.O.
(Z)
-------
Table 20. Effects on the mysid shrimp (Mysidopsis bahia) during the 6/21-6/28/88 laboratory exposure to
ambient waters from the Calcasieu River. Effects measured are growth (weight) and survival. Percent
ambient sample is the mean percent of sample in the test treatment over seven days; the remainder of
the treatment water consisted of the Narragansett brine required to adjust sample salinity.
FINAL MEAN MEAN MEAN
STATION SURVIVAL MEAN DRY VT/ TEMPERATURE SALINITY D.O.
(% SAMPLE) (%) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
BRINE + DI
87.5
0.163
0.027
26.1
+ 0.3
21.0
+
0.8
5.8
0.5
3 (95)
59.4
b
0.154
+
0.039
26.2
+ 0.3
21.0
+
0.9
5.6
+
0.7
y-s
00
00
w
87.5
0.169
+
0.014
26.2
~ 0.3
21.0
+
0.9
5.6
+
0.5
5 (90)
56.3
b
0.172
+
0.033
26.2
+ 0.4
21.0
+
0.9
5.7
+
0.5
6 (90)
71.9
0.175
+
0.050
26.2
+ 0.3
21.0
+
1.0
5.6
+
0.6
8 (91)
84.4
0.210
+
0.037
26.0
+ 0.4
21.0
1.0
5.9
+
0.6
9 (94)
75.0
0.163
+
0.030
26.0
+ 0.5
20.0
0.8
5.7
+
0.5
11 (94)
59.4
0.156
+
0.038
25.8
+ 0.5
21.0
+
0.7
5.7
+-
0.6
12 (93)
78.1
0.155
+
0.036
25.8
+ 0.7
21.0
4-
0.8
5.8
+
0.5
13 (100)
85.7
0.132
+
0.028 a,b
25.8
+ 0.7
23.0
+
1.6
5.9
+
0.5
15 (93)
84.4
0.132
+
0.031 a
25.8
+ 0.7
20.0
+
0.8
5.8
0.4
17 (87)
75.0
0.147
+
0.029
25.7
+ 0.5
21.0
+
0.8
6.1
+
0.6
18 (80)
65.6
b
0.183
+
0.037
25.7
+ 0.5
21.0
f
0.9
5.8
+
0.5
(SITE CONTROL)
a Significantly lower than the site control,
b Significantly lower than the brine + DI control.
-------
Table 21. Effects on the mysid shrimp (Mysidopsis bahia) during the 6/28-T-7/5/88 laboratory exposure to
ambient waters from the Calcasieu River. Effects measured are growth (weight) and survival. Percent
ambient sample is the mean percent of sample in the test treatment over seven days; the remainder of the
treatment water consisted of the Narragansett brine required to adjust sample salinity.
FINAL MEAN MEAN MEAN
STATION SURVIVAL MEAN DRY WT/ TEMPERATURE SALINITY D.O.
(X SAMPLE) (Z) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
BRINE + DI
87.5
0.155
+
0.031
25.9
~
0.3
21.0
+
0.8
6.0
+
0.2
18 (80)
(SITE CONTROL)
81.3
0.165
+
0.011
25.6
+
0.7
22.0
+
0.6
5.8
0.4
19 (82)
40.6
a, b
0.168
+
0.034
25.7
+
0.8
21.0
+
0.6
5.9
+
0.4
20 (82.5)
0.0
a h
25.9
X
0.2
21.0
0.9
6.1
0.4
Ay U
T
-
21 (84) c
15.6
a,b
0.097
+
0.050
25.7
0.8
21.0
+
1.0
6.0
0.4
23 (90)
93.8
0.172
+
0.022
25.8
+
0.4
22.0
+
0.8
5.9
~
0.3
24 (94.5)
93.8
0.165
~
0.024
25.7
+
0.4
22.0
+
0.3
5.9
0.5
25 (87.5)
90.6
0.146
+
0.033
25.6
ป+
0.4
21.0
+
0.9
5.9
+
0.4
26 (96)
93.8
0.132
0.038 a
25.7
+
0.4
21.0
+
0.7
6.0
+
0.4
27 (96)
96.9
0.158
+
0.014
25.5
+
0.5
21.0
+
0.9
6.0
ฆf
0.5
30 (96.5)
93.8
0.181
+
0.056
25.4
+
0.7
21.0
+
0.9
6.0
+
0.5
31 (97)
81.3
0.174
~
0.027
25.3
+
0.8
21.0
+
0.8
6.1
+
0.5
34 (98)
75.0
0.163
+
0.028
25.4
+
0.7
21.0
+
0.7
6.0
+
0.4
a Significantly lower than the site control,
b Significantly lower than the brine + DI control.
c Weight data were not analyzed since survival effects were observed.
-------
Table 22. Effects on inland silversides (Menidia beryllina) larvae during the 7-day laboratory
exposure to effluents from PPG Industrial and Vista Chemical during the Calcasieu River study. Effects
measured are growth (weight) and survival.
FINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY WT/ TEMPERATURE SALINITY D.O.
(%) (X) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
PPG IND. 001
BRINE + DI
97.8
+
3.8
1.036
+
0.06
25.5
1.4
20.5
+
0.5
5.9
0.5
6.0
93.3
+
6.7
1.092
~
0.07
25.4
+
1.3
20.1
+
0.3
5.9
+-
0.6
12.0
97.8
+
3.8
1.107
+
0.03
25.3
+
1.3
20.1
0.3
6.0
+
0.6
24.0
95.6
3.8
1.149
+
0.05
25.1
1.6
20.0
+
0.2
6.0
+
0.7
48.0
97.8
+
3.8
1.123
+
0.04
25.1
+
2.0
20.0
+
0.0
6.0
+
0.8
96.0
97.8
3.8
1.090
+
0.04
24.9
+
2.7
19.9
+
0.3
6.0
+
0.8
VISTA CHEM. 001
BRINE + DI
100.0
0.0
0.824
+
0.13
24.9
+
1.6
21.2
+
1.0
6.5
+
0.5
6.0
91.1
+
10.2
0.650
~
0.09
a
25.3
+
0.8
21.1
+
0.8
6.3
0.4
11.0
93.3
7.9
0.725
+
0.08
25.5
+
0.7
20.9
+
0.7
6.6
+
0.5
21.0
93.3
+
11.5
0.653
~
0.03
a
25.4
0.9
21.1
+
0.7
6.4
+
0.5
42.0
86.7
+
13.3
0.578
+
0.02
a
25.3
+
0.8
21.3
+
0.6
6.3
+
0.6
83.0
82.2
+
7.9 a
0.335
+
0.05
a
25.2
+
1.1
20.8
+
0.6
6.0
0.7
a Significantly lower than the brine + DI control.
-------
Table 23. Effects on inland silversides (Menidia beryllina) larvae during the 7-day laboratory
exposure to effluents from Olin Corp. outfalls 001 and 010 during the Calcasieu River study. Effects
measured are grovth (weight) and survival.
FINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY WT/ TEMPERATURE SALINITY D.O.
(X) (X) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
OLIN 001
BRINE + DI
93.3
0.0
1.118
0.02
25.9
+
1.0
20.4
+
0.5
6.2
+
0.4
5.0
95.6
+
7.7
1.106
~
0.02
26.1
+
0.6
20.2
+
0.4
6.0
+
0.5
10.0
100.0
~
0.0
1.081
4-
0.11
26.1
+
0.6
20.2
+
0.4
6.3
+
0.5
20.0
97.8
3.8
1.058
ฆซ-
0.04
26.0
+
O.b
20.4
+
0.5
5.9
+
0.6
40.0
33.3
+
6.7
atb
0.605
0.05
25.9
+
0.8
20.8
+
0.5
6.3
+
0.5
81.0
n n
0.0
a
26.0
0.7
21.4
0.7
6.0
*
0.7
v U
*r
T
"t*
OLIN 010
BRINE + DI
91.1
+
3.8
1.062
+
0.04
26.2
|
+
0.5
20.0
+
0.9
6.2
+
0.4
6.0
97.8
*ฆ
3.8
1.168
4*
0.09
26.3
0.6
20.0
+
0.2
5.8
0.6
11.0
97.8
+
3.8
1.013
+
0.04
26.3
0.6
20.0
+
0.0
5.8
+
0.6
22.0
0.0
+
0.0
a
26.0
+
0.0
20.2
+
0.4
6.3
+
0.2
45.0
0.0
0.0
a
26.0
+
0.0
20.0
+
0.0
5.7
+
0.5
89.0
0.0
+
0.0
a
26.1
~
0.2
20.2
+
0.4
4.0
+
0.4
a Significantly lower than the brine + DI control.
b Weight data were not analyzed at this and higher concentrations since survival effects were observed.
-------
Table 24. Effects on inland silversides (Henidia beryllina) larvae during the 7-day laboratory
exposure to effluents from Olin Corp. outfall 028 and Conoco, Inc. during the Calcasieu River study.
Effects measured are growth (weight) and survival.
PINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY WT/ TEMPERATURE SALINITY D.O.
(X) (Z) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
OLIN 028
BRINE + DI
93.3
+
11.5
1.120
+
0.06
26.1
+
0.6
20.3
+
0.5
6.2
+
0.3
5.0
100.0
+
0.0
0.994
+
0.07
26.0
+
0.7
20.1
+
0.3
6.1
+
0.6
10.0
93.3
+
6.7
1.048
+
0.01
26.0
+
0.7
20.1
+
0.3
6.4
+
0.4
20.0
86.7
~
1.2
1.094
+
0.09
25.9
+
0.7
20.2
+
0.4
6.2
+
0.6
40.0
93.3
+
6.7
1.086
+
0.05
26.0
+
0.7
20.1
+
0.3
6.5
0.4
80.0
86.7
18.6
1.085
+
0.09
26.0
+
0.6
20.3
+
0.8
6.1
ฆf
0.5
CONOCO, INC.
BRINE + DI
001
100.0
~
0.0
1.244
0.02
26.3
+
0.6
21.0
1.1
6.2
0.5
5.0
95.2
+
4.1
0.841
+
0.02 a
26.3
+
0.7
20.6
+
0.7
6.2
+
0.4
10.0
95.2
+
8.2
0.622
+
0.01 a
26.2
+
0.7
20.6
+
0.5
6.2
+
0.5
20.0
40.5
+
21.8 a,b
0.333
+
0.04
26.3
+
0.7
20.5
+
0.5
6.2
+
0.5
41.0
0.0
A
0.0 a
26.5
0.8
20.3
0.5
5.9
0.9
V
+
+
82.0
0.0
t
0.0 a
27.0
0.0
20.2
0.4
5.6
0.4
~
+
+
4-
a Significantly lower than the brine ~ DI control.
b Weight data not analyzed at this and higher concentrations since survival effects were observed.
-------
Table 25. Effects on inland silversides (Henidia beryllina) larvae during the 7-day laboratory
exposure to effluents from Citgo Petroleum outfalls 001 and 003 during the Calcasieu River study.
Effects measured are growth (veight) and survival.
FINAL MEAN MEAN MEAN
EFFLUENT SURVIVAL MEAN DRY WT/ TEMPERATURE SALINITY D.O.
(X) (%) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
CITGO 001
BRINE + DI
100.0
+
0.0
1.282
+
0.05
26.4
+
0.8
20.9
+
0.9
6.1
+
0.4
5.0
97.6
+
4.1
1.371
+
0.04
26.3
+
0.8
20.4
+
0.5
6.2
+
0.5
10.0
100.0
+
0.0
1.297
+
0.04
26.3
+
0.8
20.5
+
0.5
6.3
+
0.5
20.0
100.0
+
0.0
1.289
+
0.02
26.3
+
0.8
20.5
+
0.5
6.3
+
0.6
40.0
88.1
+
4.1 a
1.325
+
0.06
26.3
+
0.8
20.8
+
0.4
6.2
+
0.6
80.0
95.2
+
4.1
1.195
+
0.08
26.2
+
0.7
20.9
+
0.8
6.0
4-
0.6
CITGO 003
BRINE + DI
97.6
+
4.1
1.318
0.07
26.4
+
0.7
21.0
+
1.0
6.2
0.4
5.0
95.2
+
8.2
1.278
+
0.05
26.4
+
0.6
20.5
+
0.5
6.2
+
0.5
10.5
97.6
+
4.1
1.179
0.04 a
26.4
+
0.7
20.4
~
0.5
6.3
+
0.4
21.0
100.0
+
0.0
0.993
+
0.10 a
26.3
+
0.7
20.3
+
0.5
6.2
+
0.4
42.0
88.1
+
4.1
0.589
0.03 a
26.3
+
0.7
20.2
+
0.4
6.0
ฆf
0.3
84.0
0.0
+
0.0 a
26.6
0.6
20.2
+
0.4
5.3
+
0.4
a Significantly lower than the brine + DI control.
-------
Table 26. Effects on inland silversides (Menidia beryllina) larvae during the 6/21-6/28/88 laboratory
exposure to ambient vaters from the Calcasieu River. Effects measured are growth (weight) and survival.
Percent ambient sample is the mean percent of sample in the test treatment over seven days; the remainder
of the sample consisted of the Narragansett brine required to salinity adjust samples.
PINAL MEAN MEAN MEAN
STATION SURVIVAL MEAN DRY UT/ TEMPERATURE SALINITY D.O.
(X SAMPLE) (%) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
3 (95)
93.3
+
6.7
1.173
0.07
25.9
+
0.6
20.3
+
0.5
5.8
0.7
4 (88)
97.8
+
3.8
1.138
~
0.02
25.9
+
0.9
20.3
+
0.5
5.7
+
0.5
5 (90)
95.6
+
7.7
1.129
+
0.02
25.8
+
0.7
20.0
~
0.0
5.8
+
0.4
6 (90)
95.6
3.8
1.128
+
0.10
25.7
+
0.6
20.1
+
0.3
5.7
+
0.5
8 (91)
97.8
+
3.8
1.128
~
0.05
25.5
+
1.2
20.2
+
0.4
5.9
+
0.6
9 (94)
97.8
~
3.8
1.139
~
0.05
25.7
+
1.3
20.0
+
0.0
5.8
+
0.8
11 (94)
97.8
+
3.8
1.168
~
0.04
25.7
+
1.2
20.0
+
0.0
5.9
0.9
12 (93)
95.6
+
3.8
1.148
+
0.04
25.5
~
1.0
20.3
~
0.5
5.9
+
0.8
13 (100)
95.6
+
7.7
1.124
0.04
25.5
+
1.6
22.0
0.0
5.9
+
0.7
15 (93)
100.0
0.0
1.135
+
0.06
25.4
+
1.4
19.9
+
0.3
5.9
+
0.8
17 (87)
97.8
+
3.8
1.137
0.01
25.5
+
1.1
20.0
+
0.0
5.9
0.7
18 (80)
100.0
+
0.0
1.125
~
0.04
25.6
+
1.1
20.4
+
0.5
6.0
+
0.8
(SITE CONTROL)
-------
Table 27. Effects on inland silversides (Menidia beryllina) larvae during the 6/28-7/5/88 laboratory
exposure to ambient vaters from the Calcasieu River. Effects measured are growth (weight) and survival.
Percent ambient sample is the mean percent of sample in the test treatment over seven days; the
remainder of the treatment water consisted of the Narragansett brine required to salinity adjust
the sample.
FINAL MEAN MEAN MEAN
STATION SURVIVAL MEAN DRY WT/ TEMPERATURE SALINITY D.O.
(X SAMPLE) (Z) INDIVIDUAL (mg) (ฐC) (ppt) (mg/1)
18 (80)
91.1
+
3.8
0.869
+
0.09
25.3
+
1.2
21.0
1.0
6.5
+
0.6
(SITE CONTROL)
19 (82)
97.9
3.6
0.665
+
0.03 a
25.4
+
1.2
21.0
1.0
6.3
+
0.8
20 (82.5)
2.2
+
3.8 a
0.188
0.00
25.4
+
0.8
21.1
+
1.2
6.3
ฆh
0.6
21 (84)
51.1
+
36.7
0.194
+
0.05 a
25.2
+
1.2
21.3
+
1.0
6.4
+
0.6
23 (90)
93.3
6.7
0.824
+
0.11
25.2
+
1.0
21.4
+
1.4
6.4
+
0.5
24 (94.5)
95.6
+
7.7
0.871
+
0.04
25.3
+
0.9
21.6
+
0.9
6.3
+
0.6
25 (87.5)
95.6
+
3.8
0.842
+
0.05
25.2
+
i-
1.2
21.0
0.7
6.5
' +
0.7
26 (96)
96.7
+
4.7
0.798
~
0.00
24.9
+
1.4
21.2
+
1.0
6.3
+
0.4
27 (96)
95.6
3.8
0.818
+
0.14
25.2
+
1.2
21.0
1.3
6.3
+
0.3
30 (96.5)
88.9
+
3.8
0.805
0.06
25.1
1.2
20.6
+
0.7
6.3
+
0.5
31 (97)
93.3
+
0.0
0.798
0.04
25.1
+
1.1
20.4
+
0.7
6.4
+
0.5
34 (98)
91.1
+
7.7
0.859
+
0.06
25.3
+
1.2
20.5
+
0.7
6.3
+
0.7
a Significantly lower than the site control.
-------
Table 28. Effects of low salinity and cold storage on the toxicity
of sediments to Ampelisca abdita. The salinity experiment was
performed on uncontaminated sediments from the Narrow River, RI,
while the storage experiment was performed on sediment from from
the Calcasieu River. No significant differences in mortality were
evident between the two salinities tested or between the stored and
fresh sediments.
Treatment
Mean
Percent Survival
High (30ฐ/oo)
Salinity
99.3
Low (7ฐ/oo)
Salinity
98.0
Site 1
Fresh
76.0
Stored 7 weeks
at 4ฐC
94.4
Site 2
Fresh
7.0
Stored 7 weeks
at 4ฐC
17.8
-------
Table 29. Effects of sediments from the Calcasieu River on the
survival of Ampelisca abdita, test 1. Sediments for this test were
collected on 6/20 and tested on 6/22. Controls were from Long
Island Sound and Calcasieu River Station 18. Temperature during
the test was 20.5C + 0.5, Salinity was 30.4 + 0.5 parts per
thousand, and dissolved oxygen was >75% saturation.
Mean
Station Percent Survival
Control 96.7
(Long Island)
1
O
o
a
2
6.7
a
4
68.9
a
5
2.2
a
6
3.3
a
7
1.1
a
11
0.0
a
12
30.0
a
15
95.6
18
98.9
(SITE CONTROL)
a Significantly lower than the Long Island and Site
controls.
-------
Table 30. Effects of sediments from the Calcasieu River on the survival of
Ampelisca abdita, test 2. Samples from Stations 8-17 were collected 6/20
ana sediments from Stations 18 - 24 were collected 6/27. All samples were
tested on 7/8. Controls were from Long Island Sound, Calcasieu River
Station 18, and a low salinity control from Long Island Sound. Temperature
during the test was 20.8C + 0.5, salinity was 30.9 + 0.8 parts per thousand,
and dissolved oxygen was >75% saturation.
Station
Mean Percent
Survival
Long Island Sound Control
95.6
Low salinity Control
97.8
3A
71.1
a
3B
72.2
a
3C
36.7
a
8
76.7
9
60.0
a
10
0.0
a
13
95.6
14
90.0
16
94.4
17
97.8
18 (Site control)
96.7
19
1.1
a
20
0.0
a
21A
0.0
a
21B
0.0
a
22
0.0
a
23
0.0
a
24
90.0
a Significantly lower than the Long Island Sound,
low salinity, and site controls.
-------
Table 31. Effects of sediments from the Calcasieu River on the
survival of Ampelisca abdita, test 3. Samples were collected 6/27
and tested 7/2b. Controls were Long Island Sound sediment,
Calcasieu River Station 18, and a low salinity control from Long
Island Sound. Temperature during the test was 20.2C + 0.7,
salinity was 30.4 + 0.7 parts per thousand, and dissolved oxygen
was >75% saturation.
Mean
Station Percent Survival
Long Island Sound Control 100.0
Low Salinity Control 93.4
18 94.4
(Site Control)
25
8.9 b
26
92.2 a
27
95.6
28
94.4
29
51.1 b
30
55.6 b
31
77.8 0.
32
82.2 a
33
91.1 a
34
98.9
35
95.6
a Significantly lower than the Long Island Sound control,
b Significantly lower than the Long Island Sound, low
salinity, and site controls.
-------
Table 32. Summary of results of Calcasieu River effluent tests using Arbacia punctulata, Hysidopsis
bahia, and Menidia beryllina. All results are given as no effect concentrations (NOECs) and
lowest observed effect concentrations (LOBCs) for the most sensitive endpoint for each species and
effluent.
Effluent
A. punctulata
NOEC LOEC
M. bahia
Endpoint
NOEC
LOEC
M. beryllina
Endpoint
NOEC LOEC
PPG IND.
VISTA CHEM.
OLIN 001
7/06
7/11
OLIN 010
OLIN 028
CONOCO, INC.
7/11
7/13
CITGO 001
CITGO 003
7/11
7/13
>85.42
4.62 9.1*
<4.42 4.42
17.9% 35.72
>83.32
>70.02
17.92
8.92
>70.02
17.92
8.92
35.72
17.92
Survival/
Grovth
Survival
Survival
Grovth
Survival/
Grovth
Grovth
Grovth
Survival
>96.02
<5.02
5.02
5.02
10.02
6.02 11.02
>80.02
<5.02 5.02
20.02 40.02
10.52 21.02
Survival/
Grovth
Grovth
Survival
Survival
Survival/
Grovth
Grovth
Survival/
Grovth
Grovth
>96.02
11.02 21.02
20.02 40.02
11.02 22.02
>80.02
<5.02
>80.02
5.02
5.02
10.52
35.72
17.92
-------
Table 33. Summary of effects of Calcasieu River ambient waters on Arbacia punctulata, Hysidopsis bahia
and Menidia beryllina and effects of Calcasieu sediments on Ampelisca abdita. xx = effect,
NE = no effect, = not tested, NA = not analyzed due to survival effects.
H. bahia M. beryllina
Station /L punctulata A. abdita Survival Growth Survival Growth
1
XX
2
XX
3
NE
XX
XX
NA
NE
NE
4
XX
XX
NE
NE
NE
NE
5
XX
XX
XX
NA
NE
NE
6
NE
XX
NE
NE
NE
NE
7
XX
8
XX
NE
NE
NE
NE
NE
9
XX
XX
NE
NE
NE
NE
10
XX
i
11
XX
XX
NE
NE
NE
NE
12
XX
XX
NE
NE
NE
NE
13
XX
NE
NE
XX
NE
NE
14
NE
15
XX
NE
NE
XX
NE
NE
16
NE
17
XX
NE
NE
NE
NE
NE
18
XX
NE
XX
NA
NE
NE
-------
Table 33, cont. Summary of effects of Calcasieu River ambient waters on Arbacia punctulata, Hysidopsis
bahia and Menidia beryllina and effects of Calcasieu sediments on Ampelisca abdita. xx = effect,
NE = no effect, ป not tested, NA = not analyzed due to survival effects.
H. bahia M. beryllina
Station punctulata A. abdita Survival Growth Survival Growth
19
XX
XX
XX
NA
NE
XX
20
NE
XX
XX
NA
XX
NA
21
NE
XX
XX
NA
NE
XX
22
XX
23
XX
XX
NE
NE
NE
NE
24
NE
NE
NE
NE
NE
NE
25
XX
XX
NE
NE
NE
NE
26
XX
NE
NE
XX
NE
NE
27
XX
NE
NE
NE
NE
NE
28
NE
29
XX
30
XX
XX
NE
NE
NE
NE
31
XX
NE
NE.
NE
NE
NE
32
NE
33
NE
34
XX
NE
NE
NE
NE
NE
35
_
NE
__
-------
APPENDIX A:
Ampelisca abdita TEST METHOD
-------
Introduction
The benthic amphipod Ampelisca abdita has been shown to be sensitive to a
variety of anthropogenic materials in the marine environment. Scott et al.
(1983) developed acute toxicity test methods with A. abdita in experiments
with copper, cadmium, and arsenic. The amphipod exhibited a good dose
response to these materials, and showed a sensitivity range comparable to that
of other organisms tested. When exposed to dredged material from Black Rock
Harbor, Connecticut, in the solid phase, Ampelisca abdita was the most
sensitive of 11 species of fish and invertebrates tested (Rogerson et al.
1985). This material was contaminated primarily with polyaromatic
hydrocarbons and heavy metals. At a concentration of 5 mg/1 suspended Black
Rock Harbor sediment, growth, and consequently sexual maturation, were
delayed, and effects were seen in the laboratory population structure (Scott
and Redmond, in press). A. abdita also showed sensitivity to a series of
sediments from New Bedford Harbor, Massachusetts, which were heavily
contaminated with polychlorinated biphenyls and heavy metals (Scott et al. in
prep.).
In addition to its sensitivity, Ampelisca abdita is a useful organism for
toxicity tests because of its wide availability and ecological importance. A.
abdita is a euryhaline amphipod found from the intertidal zone to depths of
60m, ranging from Maine to southcentral Florida and the eastern Gulf of Mexico
(Bousefield 1973). It has also been introduced into San Francisco Bay
(Nichols and Thompson 1985). A tube-dwelling amphipod which constructs a
soft, upright membranous tube 3 to 4 cm long in surface sediments, A.abdita is
a particle feeder, ingesting either surface-deposited particles or particles
in suspension. Where large communities of Ampelisca are present, the patches
of amphipod tubes may affect the benthic community structure by creating a
-------
topographically complex microhabitat and by trapping suspended material
(Mills 1967). This amphipod is also a common food source for bottom fish
(Richards 1963).
Materials and Methods
Collection and Holding
Ampelisca were collected from tidal flats in the Pettaquamscutt (Narrow)
River, a small estuary flowing into Narragansett Bay, Rhode Island. Sediment
containing the amphipods was transferred to the laboratory within one half
hour, and sieved through a 0.5mm mesh screen using laboratory seawater at
ambient Narragansett Bay temperature and salinity. Ampelisca were collected
with a dip net after flotation on the air/Vater interface. The amphipods were
held in the laboratory in presieved uncontaminated collection site sediment
and flowing filtered seawater, and acclimated to the assay temperature at the
rate of 1 to 2ฐC per day. During acclimation, the Ampelisca were fed, ad
libidum, the laboratory cultured diatom Phaeodactylum tricornutum.
-------
Exposure System Design
In each experiment, Ampelisca were exposed for 10 days in flowing
filtered 20ฐC seawater to control and Calcasieu River sediments in the solid
phase. Lighting was continuous to inhibit the amphipods' swimming behavior.
The system used to deliver seawater for all but the preliminary tests is
shown in Figure 1. So that it could be used in a variety of experiments, the
system was designed with a series of components which can be modified as
needed. In these tests, the system was used solely to distribute seawater
evenly to all exposure containers. Filtered seawater at the rate of 250 ml/min
entered the upper accelerator chamber continuously. This chamber periodically
emptied into a lower, larger mixing chamber, and when the mixing chamber was
full, it in turn emptied into the 4-way flow distributor below it. The
distributor emptied into the proportioning chambers and from there into the
delivery chambers. The bottom of each delivery chamber was fitted with 15
glass capillary tubes, for a total of 60 delivery ports, and was levelled so
that each tube delivered the same amount of liquid. The capillary tubes
emptied into glass delivery tubes attached to a plate beneath the delivery
chambers. The latter were not continuous with the capillary tubing to prevent
siphoning action. The delivery tubes carried the fluid to individual exposure
containers. Turnover rate for each exposure chamber was approximately 10
volume replacements per day. A smaller system, similar in operation, was used
to deliver seawater in the preliminary tests (Figure 2).
The exposure container (Figure 3) was a 900 ml glass canning jar with a
half inch overflow hole covered with 400 micron Nitex mesh. Two hundred
milliliters of control or test sediment filled the bottom of each jar, with
approximately 600 ml of overlying seawater. A tube from the delivery system
described above entered a hole in the glass cover (3.5 inch diameter glass
-------
culture dish) of the exposure jar, emptying above the water surface. Air was
delivered from the building air supply into the water column through a glass
2 ml pipette inserted through the cover opening. Air delivery ensured
acceptable dissolved oxygen concentration and circulation of the newly
delivered seawater. Contaminated aerosol particles were prevented from
leaving through the cover opening by a foam plug. The overflow water exited
the screened overflow in the exposure chamber, flowed into a water bath, and
then to the drain and treatment system. Additional 20ฐC seawater flowed to
the water bath to stabilize the temperature in the exposure containers.
System Monitoring
The flow rate of seawater entering the seawater delivery system was
checked daily, and the volume delivered to each exposure container and from
the flow distributor was measured before and after each test. Temperature in
the water bath was monitored continuously with a temperature recorder which
was checked daily against a thermometer. Salinity of the incoming seawater
was measured daily with a refractometer, and dissolved oxygen measured in the
water column of each exposure container with a LG Nester oxygen meter twice
during each assay.
Biological Design
Before use in an experiment, each sediment was pressed through a 2 mm
mesh sieve to remove large debris and potential predator organisms, and
thoroughly homogenized. (In the preliminary sediment storage experiment, the
test sediments were not press-sieved.) Material from replicate collection
jars was composited before homogenization. The day before amphipods were
-------
added to the exposure system, sediments were press-sieved, mixed, and added to
exposure containers, and the containers filled to the overflow with filtered
seawater. A petri dish attached to a glass rod was used when adding seawater
to minimize disturbance of the sediment. Exposure containers were placed in
the water bath, and air and seawater delivery begun.
The next day, amphipods were sieved from holding containers through a
0.5mm stainless steel screen using 20ฐC filtered seawater. Juvenile amphipods
were distributed sequentially into 100-ml plastic beakers containing 20ฐC
filtered seawater. After sorting, these were examined for dead or outsized
animals, which were replaced with others from the same sieved population. The
beakers were randomized, and at least one beaker of organisms preserved in 5%
buffered formalin for later measurement by use of a computerized digitizer and
camera lucida device. Air delivery in the exposure system was halted, and
one beaker of amphipods was added to each of the exposure containers in the
experiment. After one hour containers were checked, any amphipods not
burrowed into the sediment were noted, and air delivery was restarted.
Exposure containers were checked daily and the number of individuals dead,
nearly dead, on the sediment surface, and on the water surface were recorded.
The number of molts and condition of the tubes constructed were also
monitored. Dead amphipods and molts were removed.
After 10 days, the experiment was terminated and the contents of each
exposure container were sieved through a 0.5mm screen. Material retained on
the sieve was preserved in 5% buffered formalin with Rose Bengal stain for
later sorting. Recovered animals were counted, and any missing individuals
were assumed to be mortalities.
-------
Statistical Analyses
An arc sine transformation of the square root of the proportional
mortality was conducted before analysis. Within each of the three principal
tests, mortality in each station sediment was compared with that in the site
control (station 18) using a series of t-tests (Dunnett's procedure).
Differences between station replicates (samples taken at one station site to
be tested separately), between controls (station 18, performance control, low
salinity control), and between treatments in the preliminary salinity test
were determined with a one-way analysis of variance followed by Duncan's
multiple range test.
In the preliminary sediment storage experiment, Abbott's formula was
used to correct for control mortality before the arc sine transformation and
analysis by Fisher's protected least significant difference test.
Sediment for performance controls were collected from an uncontaminated
site in central Long Island Sound (40ฐ7.95'N and 72ฐ52.7'W) with a Smith-
Maclntyre grab sampler. The sediment was returned to the laboratory, press
sieved wet through a 2 mm mesh stainless steel screen, homogenized, and
stored at 4ฐC in clean glass jars until used. A.abdita has been tested many
times in this sediment and both its survival and reproduction have been good
(Scott and Redmond, in press).
-------
Literature Cited
Bousefield, E.L. 1973. Shallow water gammaridean amphipoda of New England.
Cornell University Press, Ithaca, New York.
Mills, E.L. 1967. The biology of an ampeliscid amphipod crustacean sibling
species pair. J. Fish. Res. Bd. Canada 24(2): 305-355.
Nichols, F.H. and J.K. Thompson. 1985. Persistence of an introduced raudflat
community in South San Francisco Bay, California. Mar. Ecol. Prog. Ser. 24:
83-97.
Richards, S.W. 1963. The demersal fish population of Long Island Sound. III.
Food of juveniles from a mud locality. Bull. Bingham Oceanogr. Coll. 18:
32-101.
Rogerson, P.F., Schimmel, S.C. and G. Hoffman. 1985. Chemical and biological
characterization of Black Rock Harbor dredged material. U.S. Army Engineer
Waterways Experiment Station, Tech. Rept. D-85-9.
Scott, K.J., Yevich, P.P. and W.S. Boothman. 1983. Toxicological methods using
the benthic amphipod Ampelisca abdita Mills. Contribution 576,
Environmental Protection Agency, Environmental Research Laboratory,
Narragansett, RI.
-------
Scott, K.J. and M.S. Redmond, in press. The effects of a contaminated dredged
material on laboratory populations of the tubicolous amphipod, Ampelisca
abdita. Aquatic toxicology and hazard assessment: Twelfth Symposium.
American Society for Testing and Materials, Philadelphia, PA.
Scott, K.J., Redmond, M.S. and R.J. Pruell. in preparation. The acute response
of the amphipod, Ampelisca abdita, to contaminated sediments from New
Bedford Harbor, MA.
-------
H
njuu
I
to exposure
chaabers
Figure 1. Seawater delivery system used in three 10-day experiments exposing
Aapelisca abdlta to sediaents fro* the Calcaaieu River, Louisiana. A -
seawater inflow, B - accelerator chaaber, C - mixing chaaber, 0 - flow
distributor, E - proportioning chaabers, P - delivery chaabers, 0 - capillary
tubes, H ฆ delivery tubes. Algal food was delivered via a peristaltic pup
into the flow distributor.
-------
food
pซtri difth
ond
support
rigur* 2. Seawater dalivtry systw uMd in preliminary ซxpฎriaปntซ with
Aapallgca abdita.
-------
delivery.
to drain
water
bath
oir line
foam plug
gloss tubing
culture dish
gloss jor
screened
overflow
\ sediment
Figure 3. Exposure container used in Ampclisca abdita experiments.
------- |