Document KGpYpovJ2NDz7JGX2jVbZKk1o
072S
ACUTE TOXICITY TO AQUATIC INVERTEBRATES (EASTERN OYSTER)
TEST SUBSTANCE_____________________________________________
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METHOD:_____________________________________________________
Method: Standard Practice for Conducting Static Basic Acute Toxicity
Tests with Larvae of Four Species of Bivalve Mollusks (ASTM, Draft No. 5).
Type: Acute static
GLP: No
Year completed: 1979
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Atenstainlygt.ical monitoring: Temperature, pH, salinity, and DO.
Exposure period: 48-hours
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Dilution water: Filtered natural seawater pumped from Big Lagoon, a
Gulf of Mexico estuary adjacent to the laboratory, Pensacola, FL.
Dilution water chemistry:
SLitgohctkinagn:d
NteosSt tgasilvionelniut.yti:o2n2
ppt preparation:
A primary stock solution was
prepared by adding a weighed amount of test substance to filtered seawater.
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000615
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Temperature range Salinity range (0-48
(0-48 hours): 20 hours): 22 ppt
1
C
(Tweamteprebraattuhr)e
controlled
pDHissraonlvgeed(0o-x4y8gehonurrasn):ge8.(00--488.1hours): > 67% saturation
RESULTS____________________________________________________
Nominal concentrations: Bk control, 0.6,1.0, 3.2, 5.6, and 10 mg/L.
Element values: 48-hour EC50 = 3.5 (0.5 - 22}mg/L
Element values based on nominal concentrations
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CONCLUSIONS_______________________________________________
TChoenftiedsetnscuebIsnttaenrvcael4o8f-0h.o5utroE2C250mwg/aLs. determined to be 3.5 mg/L with a 95%
SPauublm, Mittinenr:eso3tMa, C55o1m3p3any, Environmental Laboratory, P.O. Box 33331, St.
DATA QUALITY________________________________________________
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REFERENCES________________________________________________
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OTHER____________________________________________________
Last changed: 6/27/00
000616
Acute toxicity of 3M Company's Sample
7902 to embryos-larvae of eastern
oysters (Crassostrea virginica)
/=C 2*b
L R 9 7/ 3
/
& A T 7fZVS
Toxicity Test Report Submitted to 3M Company
St. Paul, Minnesota
Project Number H90-500 Report Number BP-79-8-123
EG&G, Bionomics Marine Research Laboratory Route 6, Box 1002 Pensacola, Florida 32507 August 1979
000617
1
A marine toxicity test was conducted at Bionomics Marine 'I Research Laboratory (BMRL), Pensacola, Florida, to determine the
effect of Sample 7902 on embryos-larvae of eastern oysters (Crassostrea virginica). The criterion for effect was reduction of the number of normal larvae (those which developed to the fully-shelled, straight-hinged veliger stage within 48 hours) in test concentrations as compared to the number of normal control larvae. Results of the test are expressed as a ^fl^hour EC5ft))(the concentration of Sample 7902 estimated to be effective in preventing normal development of 50% of the exposed embryos-larvae) .
Data from the test are maintained at BMRL.
MATERIALS AND METHODS
Test material ) The sample was received at BMRL on 3 July 1979, and was con
tained in a 500-milliliter (mi) NALGENE bottle labeled "3M SAMPLE ^7902^) BIOASSAYS: 96hr LC50-GRASS SHRIMP (PALAEMONETES VULGARIS);
48hr LC50-ATLANTIC OYSTER LARVAE (CRASSOSTREA VIRGINICA)." The sample was a medium orange liquid.
Concentrations are reported here as milligrams (mg) of whole test material per Jl of seawater or as parts per million (ppm) . Test animals
Oyster embryos were obtained by induced spawning of sexually mature adult oysters which had been collected from an estuary adjacent to Biloxi, Mississippi, on 20 July 1979 and maintained in flowing, unfiltered seawater at BMRL until testing began.
*
J
000618
Test water
Water used for spawning and testing was natural seawater which
was pumped from Big Lagoon, a Gulf of Mexico estuary adjacent to
BMRL. The pump intake was about 85 meters (m) offshore at a depth
of approximiitely 3m..
Seawater was pumped by a #316 stainless steel pump through
hard polyvinylchloride (PVC) pipes, through sandfilled fiberglass
filters, and through 10-micrometers (jim) pore size polypropylene
core filters into an elevated fiberglass reservoir. Water was
continuously and vigorously aerated in the reservoir and flowed
by gravity through PVC pipes into the laboratory. There it was
pumped through a 5-ym pore size polypropylene core filter and
distributed into test chambers.
-
The chemical composition of BMRL seawater is characterized
in Appendix A.
Test conditions
Methods for the 48-hour oyster embryo-larvae test were based' on ,
Standard Practice for Conducting Static Basic Acute Toxicity Tests
with Larvae of Four Species of Bivalve Molluscs (ASTM, Draft No. 5) .
Individual, sexually mature female oysters were induced to spawn by
placing them in glass chambers containing 1 l of filtered (5-pm),
26 degrees Celsius (C) seawater and increasing the water temperature
to 32C in the presence of viable sperm excised from the gonad of a
sexually mature male oyster. Fertilization occurred upon release of
the eggs into the spawning chambers and was confirmed microscopically.
Fertilization success was estimated to be 29A0%. Density of the embryos
was determined by a Sedgwick-Rafter count of a 1:10 dilution (1 mS,
embryo suspension:9 mi, seawater) from the spawning chamber.
000619
3
All concentrations and the control were triplicated. Test containers were l- glass beakers, each of which contained 900 mi, of filtered (5-ym), natural seawater. A primary stock solution was prepared by adding a weighed amount of Sample 7902 to a known volume of filtered seawater and the appropriate volumes were added to each .test container to obtain the desired test concentrations.
Each test container was inoculated with an estimated 23,400 embryos within 1 hour after fertilization and then maintained at 201C in a temperature-controlled water bath.
After 48 hours of exposure, the larvae from each container were collected in a 37-ym mesh size sieve, rinsed into a plastic bottle with 24 mi, of filtered seawater, and preserved with 1 mi, of neutralized formalin. The number of normally developed 48-hour larvae was determined by a Sedgwick-Rafter count from each tripli cate test and control container.
Percentage reduction of normal embryos was determined as follows: Number of normal 48-hour control larvae minus the number of normal 48-hour larvae
Percentage _ in each test concentration______ .________ . reduction Number of normal 48-hour control larvae x u
The test was conducted 1-3 August 1979. Statistical analyses
Each test concentration was converted to a logarithm and the corresponding percentage reduction of normal larvae was con verted to a probit (Finney, 1971) . The 48-hour EC50 and 95% con fidence limits were then calculated by linear regression.
000620
RESULTS AND DISCUSSION
4
The calculated 48-hour EC50 for embryos-larvae of eastern oysters exposed to Sample 7902 in static, unaerated seawater was ^.5 ppm with 95% confidence limits of 0.5-22 ppm^ Reduction of embryos-larvae which developed normally to the straight-hinged veliger stage after 48 hours was from 12% in 0.6 ppm to 74% in 10 ppm (Tables 1 and 2).
Measured concentrations of dissolved oxygen remained 67% of saturation and the pH was from 8.0-8.1 after 48 hours of exposure.
V*
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000621
5 REFERENCES American Society for Testing and Materials Committee E-35 on Pesticides. August 1978. Standard Practice for Conducting Static Basic Acute Toxicity Tests with Larvae of Four Species of Bivalve Molluscs. Draft No. 5. Finney, D.J. 1971. Probit Analysis. Cambridge University Press, London. 333 p.
)
V'
000622
TABLE 1.
Toxicity of 3M Company's sample 7902 to embryoslarvae of eastern oysters (Crassostrea virginica) exposed for 48 hours in static, unaerated seawater. The criterion for effect was the reduction of the number of normal larvae in test concentrations as compared to the number of normal control larvae. Salinity was 22 /oo and temperature, 201C.
6
Nominal concentration
(mg/l }ppm)
Percentage reduction of normal 48-hour larvae^
Control
0.6 12
1.0 31
3.2 44
5.6 61
10 74
) ----------------------------------------------------------------------------------------------------------------------------------------
Number of normal 48-hour control larvae minus the number of normal 48-hour larvae Percentage _ in each test concentration____________reduction Number of normal 48-hour control larvae
)
000623
TABLE 2.
Calculated number of normal eastern oysters (Crassostrea virginica) larvae following 48 hours of exposure to 3M Company *s Sample 7902 in static, unaerated seawater. The numbers were based on Sedgwick-Rafter counts. Initial inoculum was 23,400. Salinity was 22 /oo and temperature 201C.
o
H
Nominal concentration
(mq/2,;ppm) Control 0.6
3.2 5.6 10
Rep A 19,380 15,390 13,822 12,112
8,408 5,771
Number of normal larvae Rep B Rep C Mean SDS
22,515 18,382 20,092 2,157
18,098 19,238 17,575 1-,976
11,970 15,818 13,870 1,924
8,978 12,825 11,305 2,046
6,484
8,764 7,885 1,226
3,919
6,270 5,320 1,239
aStandard deviation.
000624
APPENDIX A
Results of Chemical Analyses for Routine Characterization of Selected Chemical
Constituents in Bionomics Marine Research Laboratory Seawater
Constituent
Concentration (mg/Jt; ppm) 30 January 1979 14 June 1979
Arsenic Cadmium Chromium Copper Mercury Nickel Zinc Lead Total Phosphate as P Ammonia Nitrogen as N Nitrate Nitrogen as N Nitrite Nitrogen as N Total Petroleum Hydrocarbons Sulfides : Pesticides Polychlorinated Biphenyls
<0.001 0.002 0.0075 0.023 0.0007 0.02 0.05
<0.001 _ <0.02
0.42 <0.01 <0.01 <5.0 <1.0
None detected3 None detected*5
0.006 <0.01 <0.01 <0.01 <0.0005 <0.01 <0.02 <0.02 <0.02
0.14 <0.01 <0.01
< 5 . 0 C-
<1.0
None detected None detected
Water samples were collected from Bionomics Marine Research Laboratory seawater system after the mixing station in the wet lab.
aPesticides:
BHC, lindane, heptachlor, heptachlor eppxide, aldrin,dieldrin, endrin, perthane, DDE, tfDE (DDD), DDT, methoxychlor, endosulfan, strobane, toxaphene, kelthane, and chlordane all <0.005
vg/l.
kpolychlorinated Biphenyls: Aroclor 1016, 1232, 1248, 1260, 1221, 1242, and 1254 all <0.05 yg/A.
cPetroleum hydrocarbon sample collected 10 July 1979.
000625
PREPARED BY:
Terry A. Hollister
8
AUDITED BY REVIEWED BY: APPROVED BY:
G . Scott Ward Quality Assurance Unxt
Peter Shuba, Ph.D. Project Coordinator Rod Parrish
000G26
