ML20094H636
| ML20094H636 | |
| Person / Time | |
|---|---|
| Site: | Perry  |
| Issue date: | 10/31/1995 |
| From: | Mcculloch W, Sohn V EA ENGINEERING, SCIENCE & TECHNOLOGY, INC. |
| To: | |
| Shared Package | |
| ML20094H598 | List: |
| References | |
| 2141, NUDOCS 9511140278 | |
| Download: ML20094H636 (39) | |
Text
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. RESULTS OF ACUTE TOXICITY TESTS WITH Daphnia magna AND Pimephalespromelas ON SAMPLES FROM CALGON CORPORATION Preparedfor:
Calgon Corporation Prepared by:
g EA Engineering, Science, and Technology, Inc.
15 Loveton Circle Sparks, Maryland 21152 October 1995 EA Project 70003.10 Report Number 2141 951114027d 951103 PDR ADDCK 05000440 P PDR
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1 RESULTS OF ACUTE TOXICITY TESTS :
s WITH Daphnia magna AND Pimephales promelas l ON SAMPLES FROM CALGON CORPORATION Preparedfor: .
calgon corg. tion Prepamt by:
EA hpW, Science, and Technology, Inc.
15 Ieveton Circle Sparks, Maryland 21152 l
Jo Oefe k /995 l I VirgidA. Sohn Date l Pro.imt Manager M '
30 b dro k \ % f Wayne L. McCulloch ' Date Senior Scientist October 1995 Report Nanber 2141
I. INTRODUCTION At the request of Calgon Corporation, EA Engineering, Science, and Technology (EA) performed acute toxicity screening tests with Daphnia magna (water flea) and Pimephales promelas (fathead minnow) on samples from Perry Nuclear Power Plant, North Perry, Ohio.
Each species was exposed to undiluted (100 percent) (1) influent, (2) effluent grab sample i
collected prior to chemical treatment for zebra mussels and (3) effluent composite sample
. collected during chemical treatment.
Test results, test organism source and age, reference toxicant data, and water quality ranges measured during the tests are .ummarized on pages 3-6. Sample collection and initial water I quality data are reported in Table I. Test protocols, copies of raw data sheets, and the Report Quality Assurance Record are included as Attachments I, II, and III, respectively.
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RESULTS !
1 The results of the D. magna acute toxicity screening tests performed on the samples collected from Perry Nuclear Power Plant, North Perry, Ohio are presented on page 4. At 48 hours5.555556e-4 days <br />0.0133 hours <br />7.936508e-5 weeks <br />1.8264e-5 months <br />, there was 100 percent survival of organisms exposed to the influent sample, the effluent grab sample collected prior to zebra mussel treatment and the composite effluent sample collected during treatment. Survival in the laboratory control was also 100 percent.
The results of the Pimephalespromelas tests are summarized on page 6. The three undiluted samples from Perry Nuclear Power Plant had 100 percent survival after 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> of exposure.
The laboratory control had only 85 percent survival, which is less than the minimum of 90 percent for acceptable control survival based EA quality control criteria. The results of the most recent reference toxicant test performed on the in-house cultured stock of fathead l minnows was within an acceptable range, based on EA's Aquatic Toxicology Control Chart.
In our best professional judgment, the poor laboratory control survival does not affect the integrity of the results of the tests performed on the three Perry Nuclear Power Plant samples.
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SUMMARY
OF SAMPLFJTEST INFORMATION i Test: 48-hour Daphnia magna Static Acute Toxicity Test Client Name: Calgon Corporation Sample Site: Perry Nuclear Power Plant
. Test Procedure: EA Protocol ATS-SAI-DS-04 Acute 48-hour assay with Daphnia Sample
Description:
Influent; Emuent grab prior to treatment; Emuent composite during tatatment EA Accession Number: ATS-1088, AT5-1989, AT5-1090 EA QC Test Number: TN-95-678 Test initiation time and date: 1330, 6 October 1995 Test completion time and date: 1313, 8 October 1995 Control Water: Dechlorinated tap water Orennimm Int Informatinn I.ot Number: Not Applicable Source: EA's Culture Facility, Sparks, Maryland Age: <24 hours Reference Toriennt (SDS) Test Informatinn Reference Toxicant: Sodium dodecyl sulfate EA QC Test Number: RT-09-26-95-162 Dilution Water: Dechlorinated tap water Results:
48-hour If50: 17.1 mg/L SDS Laboratory control chart negability range for 48-hour LC50:
8.7 - 19.4 mg/L SDS 3
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SUMMARY
OF SAMPLE / TEST INFORMATION (Cont.) 1 Test Organism: D. magna EA QC Test Number: TN-95-678 Test Matenal
- (100 nercent concentration) Percent Survival at 48 Hours Laboratory Control 100 Influent (ATS-1088) 100 Effluent prior to treatment (AT5-1089) 100 Effluent during treatment (AT5-1090) 100 Relected Wnter Onnlity Rnnae Temperature (*C) 19.9 - 20.8 pH 7.9 - 8.6 Dissolved oxygen (mg/L) 8.0 - 8.8 Conductivity ( S/cm) 258 - 374 l
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SUMMARY
OF SAMPLFJTEST INFORMATION l
l Test: 96-hour Pimephalespromelas Static Acute Toxicity Test ,
Client Name: Calgon Corporation Sample Site: Perry Nuclear Power Plant i i
~ l Test Procedure: EA Protocol ATS-SAF-FM-04 Acute assay with fathead minr.ows (Pimephalespromelas)
Sample
Description:
Influent; Emuent grab prior to titatment; Emuent composite during treatment EA Accession Number: AT5-1088, ATS-1089, AT5-1090 EA QC Test Number: IN-95-679 Test initiation time and date: 1500, 6 October 1995 Test completion time and date: 1530,10 October 1995 Control Water: Dechlorinated tap water n
Oranninm Imt Informatinn Iot Number: FH 5-10/3 Source: EA's Culture Facility, Sparks, Maryland Age: 3 days (hatched within a 24-hour window) j Reference Tnriennt (CdCl3 Test Information i Reference Toxicant: Cadmium chloride EA QC Test Number: RT-09-16-95-158 Dilution Water: Dechlorinated tap water
. Results:
48-hour LC50: 20.8 yg/L Cd Iaboratory control chart acceptability range for 48-hour LC50:
15.2 - 55.2 g/L Cd 5
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SUMMARY
OF SAMPLFJTEST INFORMATION (Cont.)
Test Organism: P. proselas -
EA QC Test Number: TN-95-679 Test Material
. (100 nercent enne entration) Percent Survival at 96 Hours I4 oratory Control 85 "
Influent (ATS-1088) 100 Effluent prior to treatment (AT5-1089) 100 Effluent during treatment (ATS-1090) 100 Relecteel Water Onnlity Rnnsre Temperature (*C) 19.6 - 20.8 pH 7.0 - 8.2 Dissolved oxygen (mg/L) 8.5 - 9.1 Conductivity ( S/cm) 261 - 374 (a) See page 2 for discussion.
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TABLE 1 SAMPLE COLLECTION AND INITIAL WATER QUALITY DATA FOR SAMPLES FOR CALGON CORPORATION '
Sample EA Accession Type of Sample Sample Alkalinity Hardness Dew ririion Numher Sample Collection Receipt (me/L CaCOJ (me/L CACO)
Influent ATS-1088 Grab 0940,4 OCT 95 0930,6 OCT 95 80 108 Effluent prior ATS-1089 Grab 0850,4 OCT 95 0930,6 OCT 95 112 '
132 to treatment i
Effluent ATS-1090 Composite 0954,4 OCT 95 to 0930,6 OCT 95 100 132 j during treatment 2120,4 OCT 95 '
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ATTACHMENTI PROTOCOLS l
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. PROTOCOL ATS-SAI-DS-04 ACUTE ASSAYWITH Daphnia sp.
- 1. TEST OBJECTIVE To assess the toxicity of a test material to Daphnia and determ%e the LC50 or EC50 using mortality or immobilization, respectively, as the test end points.
- 2. TEST ARTICLE 2.1 Dex pGon/ Identification Unless otherwise specified, the test material is supplied by the client. Adequate chemical specifications with special refemnce to hazardous properties and storage conditions is also supplied by the client.
2.2 Methods of Synthesis In most cases the test article is an effluent sample. Information on the methods of synthesis, stability, and composition or other characteristics which define the test article are on file with the client.
- 3. EXPERIMENTAL DESIGN 3.1 Test Organisms A species of Daphnia (water fleas), as determined by project needs, is the test organism.
3.2 Source Daphnia used for acute toxicity tests are obtained from stock cultures maintained in EA's Aquatic Toxicology Culturing Facility.
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ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 1 of 9
PROTOCOL ATS-SAI-DS-04 3.3 Culturine and Holdine Conditions Daphnia cultures are maintained at 20i2 C and a 16-hour light, 8-hour dark photoperiod cycle in an environmentally controlled laboratory. Cultures are maintained in 18.9-L all glass aquaria or other appropriate container and are fed algae (Selenastrum capricornutwn) and a trout chow / yeast / cereal leaves suspension in the manner described in Weber 1991.
New cultures are initiated on a routine basis to ensure healthy, productive populations.
, Organisms from cultures producing ephippia are not used for toxicity tests. Certain regulatory or project specific objectives may require organism acclimation to the dilution water when it is different from the holding / culture water.
3.4 Are of Test Organisms at Test Initiation Neonates of known age (i.e., less than 24-hours old) are obtained for testing by segregating adult females from the mass cultures on the day before a test is initiated. On the day of the test, neonates are segregated from the parent organisms.
3.5 Dilution Water The source of dilution water is the City of Baltimore Municipal Water System. Upon entry to the laboratory, the water is passed through a high-capacity, activated-carbon filtration system to remove chlorine and other possible contaminants such as detergents, and hexane-soluble organic materials. This water source has proven safe for aquatic organism toxicity testing as evidenced by maintenance of the multigeneration Daphnia and Pimephales promelas cultures, with no evident loss of fecundity. If requested, reconstituted fresh water or an appropriate receiving water can be used as the dilution water.
3.6 Test Concentration Series
' The test concentration series consists of a minimum of five dilutions and may be determined from a prior screening of the test material (e.g., 1, 3,10, 30, and 100 percent effluent; or 6.25,12.5,25,50, and 100 percent effluent plus a control).
ATS-102 EA Engineering, Science, l and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 2 of 9
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3.7 Test Concentration Preoaration Test concentrations are prepared with Class A glassware.
3.8 Test vessels and Test Volume Test vessels are 400-ml glass beakers or 200-ml culture bowls; the final test volume of each
. replicate is 200 ml. Depending on the study requirements, other test vessels and test volumes may be used.
3.9 Test Organism Number Tests are conducted using two replicates per concentration, with ten organisms per container.
Neonates are randomly assigned to each replicate test container. More replicates can be added, if appropriate.
3.10 Test Environment The test vessels are maintained at 20 1 C or 25il C in an environmentally controlled laboratory with a 16-hour light, 8-hour dark photoperiod.
3.11 Analysis of Test Concentrations for Test Article If required, test solutions may be analyzed for verification of chemical concentrations. The analytical method and number of analyses are determined after consultation with the client.
When chemical analyses are necessary, both nominal and actual measured test solution concentrations are reported.
3.12 Test Observations
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Each test day, test organisms are observed to record the number of surviving organisms.
The study terminates after completion of the 2-day observation period. The study may be
. extended, however, at the request of the client.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 3 of 9 1
. PROTOCOL ATS-SAI-DS-04 Each sample received is analyzed for temperat'ure, conductivity, alkalinity, and hardness.
Measurements of water quality taken daily include dissolved oxygen, pH, temperature and conductivity from a minimum of one replicate of every concentration. Aliquots of effluent and receiving water may be gently aerated (100 bubbles / min) if dissolved oxygen is less than
- 4 mg/L or greater than 105% saturation. After test initiation, if the dissolved oxygen in any test chamber is less than 4 mg/L, all test chambers are gently aerated or other corrective action is taken. Analytical determinations are conducted according to APHA et al. (1992)
, and EPA (1979).
3.13 Solution Renewal (When Applicable)
When static-renewal testing is required, the test solutions are renewed at 24-hours. New test solutions are prepared on the day of renewal and dispensed according to Section 3.7. After the new solutions have reached test temperature, the test organisms are transferred from the old test vessels to the new test vessels using a wide bore pipet or glass tube. The number of live organisms is recorded. Caution is given not to stress the test organisms during transfer.
After water quality measurements (temperature, pH, dissolved oxygen, and conductivity) are completed, the old solution is discarded.
3.14 Data Analysis The LC50 or EC50 values and associated statistics are calculated using the probit, moving average, and binomial methods as described by Stephan (1977). Depending on the nature of the data, other methods may be used, including the Trimmed Speannan-Karber method, the probit approximation method of Litchfield and Wilcoxon (1949), SAS probit analysis (SAS Institute 1985) or graphical interpolation using the log concentration vs. percent mortality as described by APHA et al. (1989). The methods used are specified in the final report.
3.15 Test Accentability An individual test may be conditionally acceptable if temperature, DO, and other specified conditions fall outside specifications, depending on the degree of the departure and the
, objectives of the tests.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 4 of 9 1
PROTOCOL ATS-SAI-DS-04
- 4. FINAL REPORT The final report is prepared to contain at a minimum the following information:
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Objectives and procedures stated in the approved protocol, including any changes made to the original protocol Identity of the test article (s) by name or code number and the strength (i.e., quality / purity and a description of any pretreatment)
Source of the dilution water, its chemical characteristics, and a description of any pretreatment Test concentration series used and duration of the assay
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Water quality characteristics of dilution water and selected test concentrations during testing (pH, dissolved oxygen, temperature, etc.)
Any unforeseen circumsamces that may have affected the quality or integrity of the study Signature of the project manager, senior technical reviewer, and quality control officer, authorizing release of the report I.ocation of all archived data and the original copy of the final report at EA Items of data to be included in the report consist of experimental design and test performance; effects on general appearance of test organisms (if applicable); morbidity and mortality; presentation of water quality characteristics; and survival data.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992
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. PROTOCOL ATS-SAI-DS-04
- 5. QUALITY ASSURANCE l 5.1 Amendments to Protocol
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Amendments to the authorized protocol established by EA or by the client are made only !
after proper authorization. Such authorization is achieved by completion of the Amendment l to Protocol Form by EA after consultation with the client.
5.2 Standard Operatine Prrwwiures Unless otherwise specified, all procedures specified in the protocol are subject to detailed Standard Operating Procedures (SOPS) which are contained in the SOP manuals of the participating departments. These SOPS and protocols generally follow the types of requirements as outlined in the U.S. EPA's Good 14 oratory Practice Standards (GLPs)
(EPA 1989).
5.3 Reference Toxicant A reference toxicant test, utilizing sodium dodecyl sulfate (SDS), cadmium chloride, or another appropriate chemical is used as an internal quality check of the sensitivity of the test organisms. Testing is. conducted at least once monthly on organisms which are cultured
, in-house. The results of eac'i test are compared with historical, species-specific toxicological !
information from reference toxicant tests performed at EA, to determine if the results are within acceptable limits. Limits are established using the control charts outlined in Weber (1991).
5;4 Ouality Assurance Evaliation Studies conducteo under this protocol may be subject to internal audit by EA's Quality
, Assurance Unit. A q'tality control officer is responsible for monitoring each study to assure the client that the facilities, equipment, personnel, methods, practices, records, and controls are in conformance with EA's QC program and, if applicable, EPA's GLPs.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effecdve May 1981 Revision 4, April 1992 Page 6 of 9
PROTOCOL ATS-SAI-DS-04 5.5 Insnection by Reculatory Authorities In the event of an inspection of EA by an outside authority during the course of the study, the client whose study is being inspected will be consulted before examiners are permitted access to any of the project records or the experimental areas.
5.6 Archives Copies of project-specific records shall be transferred to the client promptly after the project is completed or as negotiated and budgeted. Original primary data are retained at EA for 5 years. Primary data include chain-of-custody records, laboratory data sheets, records, memoranda, notes, photographs, microfilm, and computer printouts that are a result of the original observations and activities of the study and which are necessary for the reconstniction and evaluation of the study report.
5.7 Location Studies are conducted at the Aquatic Toxicology Laboratory of EA Engineering, Science, and Technology, Inc. at the IAveton Office in Sparks, Maryland.
- 6. SPECIFICATIONS OF THE Daphnia ACUTE TOXICITY TEST 6.1 Basic References American Public Health Association (APHA), American Waterworks Association, Water Environment Federation.1992. Standard Methods for Examination of Water and Wastewater,18th edition or most recent version. APHA, Washington, D.C.
EA Manual ATS-102.1992. Aquatic Toxicology Studies, Quality Control and Standard
- Operating Procedures Manual. Fourth Revision. Intemal document prepared by Aquatic Toxicology Services, EA Engineering, Science, and Technology, Inc.
Litchfield, J.T., Jr. and F. Wilcoxon. 1949. A simplified method of evaluating dose-effect experiments. J. Pharm. Exp. Ther. 96:99-113.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 7 of 9
PROTOCOL ATS-SAI-DS-04 SAS Institute Inc. 1985. SAS* User's Guidei Basics, Version 5 Edition. Cary, NC: SAS Institute Inc. 1290 pp.
Stephan, C.E. 1977. Methods for calculating an LC50, in Aquatic Toxicology and Hazard
American Society for Testing and Materials, Philadelphia.
U.S. Environmental Protection Agency. 1979. Methods for Chemical Analysis of Water and Wastes. EPA-600/4-79-020. U.S. EPA, Washington.
Weber, C.I. 1991. Editor. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms. Fourth Edition.
EPA /600/4-90/027. U.S. Environmental Protection Agency. Environmental Monitoring Systems Laboratory, Cincinnati, Ohio.
U.S. EPA. 1989. Toxic Substances Control Act (TSCA); Good Laboratory Practice Standards. Title 40 CFR Part 792. Ecd. Regist. 54(158):34034-34074.
6.2 Test Soecifications Test organism: Daphnia magna or D. pulex; species to be specified in the study plan and final report Age: Less than 24 hours2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> old Temperature: 20i1 C or 25 1C Light quality: Wide-spectrum fluorescent light
, Light intensity: 50-100 f.c.
Photoperiod: 16-hour light, 8-hour dark Aeration: None, unless dissolved oxygen falls below 4 mg/L l
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Effective May 1981 Revision 4, April 1992 Page 8 of 9 l
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PROTOCOL ATS-SAI-DS-04 Dilution water: Dechlorina' ted municipal water, reconstituted water, or appropriate receiving water
, Test container: 400-ml beaker or 200 ml culture bowl Test volume: 200 ml per replicate No. of concentrations: Minimum of five test concentrations and a control No. of replicates: Two No. of animals per replicate: Ten Feeding regime: Animals will not be fed during test Test type and duration: Rangefinding--24 to 48 hours5.555556e-4 days <br />0.0133 hours <br />7.936508e-5 weeks <br />1.8264e-5 months <br /> Definitive--48 hours Endpoints: Mortality or immobilization Immobilization-defined as cessation of mevement except for minor activity of appendages Mortality--defined as cessation of all movement for a period of at least 5 seconds even when the test container is tapped or rotated, or the organism is gently prodded with glass rod
, Test acceptability: 90% or greater survival in the control solution.
ATS-102 .
EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 9 of 9
PROTOCOL ATS-SAF-FM-04 ACUTE ASSAY WITH FATHEAD MINNOWS (Pimephale's promelas)
- 1. TEST OBJECTIVE
- To assess the toxicity of a test material to Pimephales promelas and determine the 1450 or EC50 using mortality and/or immobilization, respectively, as the test endpoint (s).
, 2. TEST ARTICLE 2.1 Derri%on/ Identification Unless otherwise specified, the test material is supplied by the client. Adequate chemical specifications with special reference to hazardous pipdes and storage conditions are also supplied by the client.
2.2 Mathnde of Syntheeic In most cases the test article is an effluent sample. Information on the methods of synthesis, stability, and composition or other characteristics which define the test article are on file with the client.
- 3. EXPERIMENTAL DESIGN 3.1 Test Oreanisms
'Ihe test species is the fathead minnow, Pimephales pmmelas.
3.2 Source P. promelas used for toxicity tests are usually obtained from stock cultures maintained at EA's Aquatic Toxicology Culturing Facility. However, organisms may be obtained from a scientific organism vendor, if r+:"=ry.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 1 of 11 l
PROTOCOL ATS-SAF-FM-04 3.3 Culturine and Holdine Conditions P. promelas stock are maintained at 20 2 C and a 16-hour light, 8-hour dark photoperiod cycle in an environmentally controlled laboratory. Test organisms are maintained in 18.9-L or 30-L all glass aquaria in a recirculating system of dechlorinated municipal tap water. If
- organisms are obtained from a scientific vendor, the fish, upon receipt, are transferred to static recirculating holding tanks of an appropriate size containing dechlorinated tap water at the temperature ( 2 C) of the water in which the organisms were shipped. All fish stocks
- are examined regularly. Dead fish, or those displaying abnormal swimming behavior, discoloration, or pronounced lethargy are removed as observed, and recorded on appropriate log sheets. Fish stocks are fed a commercial fish food or Anemia nauplii a minimum of once daily. Certain regulatory or project specific objectives may require organism acclimation to the dilution water when it is different from the holding / culture water.
3.4 Are of Test Organisms at Test Initiation The age of the test organisms is dependent on the objectives of the study and the specific guidelines being followed, but in general, fathead minnows used for acute toxicity testing are 1-14 days old (with a 24-hour range in age) upon test initiation.
3.5 Dilution Water The source of dilution water is the City of Baltime e Municipal Water System. Upon entry to the laboratory, the water is passed through a Egh-capacity, activated-carbon filtration system to remove chlorine and other possible contaminants such as detergents and hexane-soluble organic materials. This water source has proven safe for aquatic organism toxicity testing, as evidenced by maintenance of the multigeneration Daphnia and fathead minnow cultures, with no evident loss of fecundity. If requested, reconstituted fresh water or other dilution water may be used.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 2 of 1I
PROTOCOL ATS-SAF-FM-04 3.6 Test Concentration Series The test concentration series consists of a minimum of five dilutions and may be determined from a prior screening of the test material (e.g., 1,3,10,30, and 100 percent effluent; or 6.25,12.5,25,50, and 100 percent effluent plus a control).
3.7 Test Concentration Preoaration
, Test concentrations are prepared with Class A glassware.
3.8 Test Vessels and Test Volume Test vessels are 2 L glass culture bowls; the final test volume is 1 L. Other test vessels may be used depending on the study's requirements.
3.9 Test Orcanism Number Tests are conducted using 2 replicates per concentration, with 10 organisms per container.
Fish are randomly assigned to each replicate test container. More replicates can be added, if appropriate.
3.10 Test Environment The test vessels are maintained at 20 1 C or 25 1 C (unless a different project-specific temperature is required) in an environmentally controlled laboratory with a 16-hour light, 8-hour dak photoperiod. Temperature within the environmental room is monitored continuously using temperature recorders.
3.11 Analysis of Test Concentrations for Test Article If required, test solutions may be analyzed for verification of chemical concentrations. The analytical method and number of analyses are determined after consultation with the client.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 3 of 11
PROTOCOL ATS-SAF-FM-04 When chemical analyses are necessary, both nominal and actual measured test concentrations are reported.
3.12 Test Observations Each test day, test organisms are observed to record the number of surviving organisms.
Dead organisms are removed when observed. The study terminates after completion of the observation period (24 to 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br />). The study may be extended, however, at the request of the client.
Each sample received is analyzed for temperature, conductivity, alkalinity, and hardness.
Measurements of water quality taken daily include dissolved oxygen, pH, temperature, and conductivity from a minimum of one replicate of every concentration. Aliquots of effluent and receiving water may be gently aerated (100 bubbles / min) if dissolved oxygen is less than 4 mg/L or greater than 105% saturation. After test initiation, if the dissolved oxygen in any test chamber is less than 4 mg/L, all test chambers are gently aerated or other correctie action is taken. Analytical determinations are conducted according to APHA et al. (1992) and EPA (1979).
At the end of the test period, a minimum of 30 surviving fish are removed from the test vessels, placed in labeled containers, and frozen for later measurements. The fish are subsequently thawed, measured to the nearest mm (standard length), patted dry, and weighed to the nearest 0.01 g. The average length and weight of the test fish are reported, along with their respective ranges and standard deviations. For fish between 1-14 days old and within a 24-hour age range, weights and lengths usually will not be measured.
3.13 Solgtion Renewal (When Applicable)
When static-renewal testing is required, the test solution is renewed daily. New solutions are prepared on the day of renewal and dispensed according to Section 3.8. After the new solutions have reached test temperature and water quality measurements (temperature, pH, dissolved oxygen, and conductivity) are completed, the solution renewal may be performed by transferring the organisms from one test chamber to another or by replacing the test ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 4 of 11 I
r PROTOCOL ATS-SAF-FM-04 solution. If the test solution replacement method is used, caution must be given not to stress the test organisms while the test chamber is si;ihoned. Usually 75 percent of the old solution is removed and replaced; however, the amount is dependent on the size of the test organisms.
3.14 Data Analysis The LC50 and/or EC50 values and associated statistics are calculated using the probit, moving average, and binomial inethods as described by Stephan (1977). Depending on the nature of the data, other methods may be used including the Trimmed Spearman-Karber
' Method, the pmbit approximation method of Litchfield and Wilcoxon (1949), SAS probit analysis (SAS Institute 1985), or graphical interpolation using the log concentration vs.
percent mortality and/or percent affected as described by APHA et al. (1992). The methods used are specified in the final report.
3.15 Test Anolicability An individual test may be conditionally acceptable if temperature, DO, and other specified conditions fall outside specifications, depending on the degree of the departure and the objectives of the tests.
- 4. FINAL REPORT The fmal report is prepared to contain, at a minimum, the following information:
- Objectives and procedures stated in the approved protocol, including any changes made to the original protocol
- Identity of the test article (s) by name or code number and their strength (i.e., quality / purity), and a description of any pretreatment
- Source of the dilution water, its chemical characteristics, and a description of any pretreatment 4
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 5 of 11
PROTOCOL ATS-SAF-FM-04
- Test concentration series used and duration of the assay
- Mean standard lengths and wet weights of test fish, the respective standard deviations, and approximate biomass loadings (g fish /L of test solution), if applicable
- Water quality characteristics (pH, dissolved oxygen, temperature, etc.) of dilution water and selected test concentrations during testing
- Any unforeseen circumstances that may have affected the quality or integrity of the study
- Signature of the project manager, senior technical reviewer, and quality control officer authorizing release of the report
- Imation of all archived data and the original copy of the final report e EA Items of data to be included in the report consist of experimental design and test performance; effects on general appearance of test organisms (if applicable); morbidity and mortality; presentation of water quality characteristics, and survival data.
- 5. QUALITY ASSURANCE 5.1 Amendments to Pie;r-wl Amendments to the authorized protocol established by EA or by the client ase made only after proper authorization. Such authorization is achieved by completion of the Amendment to Protocol Form by EA after consultation with the client.
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Effective May 1981 Revision 4, April 1992 Page 6 of 11
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PROTOCOL ATS-SAF-FM-04 5.2 Standard Ooeratint Procedures Unless otherwise speciEed, all procedures given in the protocol are subject to detailed Standard Operating Procedures (SOPS) which are contained in the SOP manuals of the participating departments. These SOPS and protocols generally follow the types of
~ requirements outlined in the U.S. EPA's Good Laboratory Practice Standards (GLPs)
(EPA 1989).
, 5.3 Reference Toxicant A reference toxicant test, utilizing sodium dodecyl sulfate (SDS), cadmium chloride, or another appropriate chemical is used as an internal quality check of the sensitivity of the test organisms. Testing is conducted at least once monthly on organisms that are cultured in-house. The results of each test are compan:d with historical, species-specific toxicological information from reference toxicant tests performed at EA, to determine if the results are within acceptable limits. Limits are established using the control charts outlined in Weber (1991).
5.4 Ouality Assurance Evaluation Studies conducted under this protocol may be subject to internal audit by EA's Quality Assurance Unit. A quality control officer is responsible for monitoring each study to assure the client that the facilities, equipment, personnel, methods, practices, records, and controls are in conformance with EA's QC program and, if applicable, EPA's GLPs.
5.5 Inspection by Reculatory Authorities In the event of an inspection of EA by an outside authority during the course of the study, the client whose study is being inspected will be consulted before examiners are permitted access to any of the project records or the experimental areas.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 7 of 11 l
i
t PROTOCOL ATS-SAF-FM-04 5.6 Archives Copies of project-specific records shall be transferred to the client promptly after the pioject is completed or as negotiated and budgeted. Original primary data are retained at EA for 5 years. Primary data include chain-of-custody records, laboratory data sheets, records, memoranda, notes, photographs, microfilm, and computer printouts that are a result of the original observations and activities of the study and which are necessary for the reconstruction and evaluation of the study report.
5.7 Location Studies are conducted at the Aquatic Toxicology 1.aboratory of EA Engineering, Science, and Technology, Inc. at the Loveton Office in Sparks, Maryland.
- 6. SPECIFICATIONS OF THE FATHEAD MINNOW ACUTE TOXICITY TEST 6.1 Basic References American Public Health Association (APHA) American Water Works Association, Water Environment Federatian. 1992. Standard Methods for Examination of Water and Wastewater,18th or most recent version. APHA, Washington, D.C.
American Society for Testing and Materials (ASTM). 1991. Standard Practice for Conducting Acute Tests with Fishes, Macroinvertebrates, and Amphibians. ASTM Designation: E729-80, Philadelphia, Pa.
EA Manual ATS-102.1992. Aquatic Toxicology Studies, Quality Control and Standard Operating Procedures Manual. Fourth Revision. Internal document prepared by Aquatic Toxicology Services, EA Engineering, Science, and Technology, Inc.
Litchfield, J.T., Jr. and F. Wilcoxon. 1949. A simplified method of evaluating dose-effect experiments. J. Pharm. Exp. Ther, 96:99-113.
ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 8 of 11
PROTOCOL ATS-SAF-FM-04 SAS Institute Inc. 1985. SAS* User's Guide: Basics, Version 5 Edition. Cary, NC: SAS Institute Inc. 1290 pp.
Stephan, C.E. 1977. Methods for calculating an LC50, in Aquatic Toxicology and Hazard Evaluation (F.L. Mayer and J.L. Hamelink, eds.), pp. 65-84. ASTM STP 634.
. American Society for Testing and Materials, Philadelphia.
U.S. Environmental Protection Agency. 1979. Methods for Chemical Analysis of Water
, and Wastes. EPA-600/4-79-020. U.S. EPA, Washington.
U.S. EPA. 1989. Toxic Substances Control Act (TSCA); Good Laboratory Practice Standards. Title 40 CFR Part 792. Ecd. Engist. 54.
Weber, C.I.1991. Editor. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms. Fourth Edition.
EPA /600/4-90/027. U.S. Environmental Protection Agency. Environmental Monitoring Systems laboratory, Cincinnati, Ohio.
6.2 Test Snecifications Test organism:
Fathead minnow (Pimephales promelas)
Temperature:
20il C or 25il C Fish age:
Dependent on test guidelines being followed, but in general,1-14 days with a 24-hour range in age Fish length:
The largest fish is not more than 50 percent longer than the smallest fish Loading ratio:
In general, test containers may not contain greater than 0.65 g/L biomass ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 9 of 11
\ - _ _ _ _ _ _ _ _ _
PROTOCOL ATS-SAF-FM-04 Aeration: None, unless dissolved oxygen falls below 4 m'g/L Light quality: Wide-spectrum fluorescent light Light intensity: 50-100 f.c.
Photoperiod: 16-hour light, 8-hour dark Dilution water: Dechlorinated municipal tap water, reconstituted fresh water, or appropriate receiving water Test containers: 2 L glass culture bowl Test volume: 1 IJreplicate No. of concentrations:
Minimum of five test concentrations and a control No. of replicates: 2 Test animals per container:
10 (if loading is exceeded, more replicates or larger test containers may be used)
Feeding regime:
Animals are not fed during tests unless fish are too small to survive % hours without feeding.
Test type and duration:
Rangefinding-24-% hours Definitive-24-96 hours ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 10 of 11 l
PROTOCOL ATS-SAF-FM-04 Endpoints: Mortality and/or Immobilization Immobilization--defined as cessation of movement except for minor activity of appendages .
Mortality--defined as cessation of opercular !
movement and inability to respond to external '
stimuli (gentle prodding with glass rod) l
' l Test acceptability: 90 percent or greater survival in the control solution l
l ATS-102 EA Engineering, Science, and Technology, Inc.
Effective May 1981 Revision 4, April 1992 Page 11 of 11
ATTACHMENTli DATA SHEETS I
1 j
l
nuun a su v..annive- = muu s m i varve Y stos uMsA GHi.tI Project No.: 70063 IC TEST Flow-through, Monswel TEST ORGANISM: Beginning Cate: 1OlIr!4I Time- tMO Cne e. enlann Test Co.,isiner. w b l. Co, on umme: u,aham ;
OC Test No.: N'N 200 O Enan. Deie. 10- & 9 5 u me. R/3 Test Volume: Scientific Nome: 0 ha Adusi. isoisted (Tim., Doies: so 5-if /4,.
Test Article- 'e Test Duration: Mr5 Lot No.: d Organisms Puned (Time, Datel: 10 -IP-1 T ' M
- Sample No. : ar<-bSilDMT.ld96 lo h Source: Accumation: # or Age O%
, Diluent: huNi# TAMGET VALUES D#vtlon No.: h[8 Temp: ."Ic71 I CWture Water T/S. 9 Transfer (12) 1*
- DO: 7'Irw+/L pH: (n- 9 Sannity: NA ALL TESTS Number of Conc. Rep. Live Organisms Dissolved Oxygen ConductMty luS/cml Temperature (*Cl pH ( . ,.O or % No , SI .iIppt) Other o 24 an 12 na o 24 am 72 na o 2a da 72 na o' 141 da 72 na o ;d as 72 m o 24 da 72 na hl A
/0 @ sh 36 4 %.I C t.I gs' M7 &
G ec /t> I() 20 33 k) 43 nrs - btf loo'I A ro 10 nA #.9 rg fr4 10 71 r.3 3x m D 'e Io Ib 00] 85 @3 338 Ars-loft loo-/ A /# lo lo @i1 *k fI P3 tct M
/0 3W St G Io 10 W.1 %5 85 3M (175-R40 1or / & ,a lo O m b.4 8t 8i 16 8.ie 3N 143
- 0) f8 b d !
10 O f'7 Instrument Number 7fp kk 1% T3 h h % h E/n kd ;
C - e n rector 4s a k &b E r.t t.m LJ %
Time $ M (3i3 til pik 13/I pit t~d> cd ip1 43o 13f% nft # 13 4 kwestigator @ Ob M W @ t:(4 7 @ 0 46 P (b E6 7" (b $(A ATS AC5 3 et 7 me:
l i
AQUATIC TOXICOLOGY BENCH SHEET SET UP (SMALL VOLUMES) !
Project No.- 7 0 3' '
Client- IW e**
[ QC Test No.. -rd-f,W
- . Date/ Time Activity Investigator i Dilutions Made h l
/H -R (12 Test Vessels Filled l
Transferred Organisms M f 3 yf Head Counts Sample No.. AE ' 10E%f 0fR,109D i
Dilution No.: 00 Md V Volume Test Test Concentration Material (ml) Final Volume O
O n..L- Q00 &
ATC~ low) l 0 0 ' l-
,,s.,e ) ior /.
400 &
its- /o9o ) 10o Y d ATS-M13 2/27/95
AQUATIC TOXICOLOGY BENCH SHEET Project No: '? D 0 0 3 ' D -
- Client: O Aleen
! QC Test No.: O - trM
., Date/Tirne Activity Investigator I JD- b -97 loso bei Do g- %
- 8. 7 PF 6o A jbg l A g -fogp 93 8.9 A Tr- lo M Y2 9. 0 MW -toto 6
e ATS-Ms2 3/10/93
__ . _ _m . _ _ . . . . . . _ _ _ _ _ . _ _ . _ . .
AQUATIC GRGANISMS ACUTE TOXICITY TEST DATA SHGET Project No.: 7 OOC)3 I U TEST: t:.ti) Flow through, Renewal TEST ORGANISM: Seginning Date: . lolle l'l5 (CdW Time: $ 00 Client: Test einer: l-l b"N Common Name: bdS Ending Date: /0NoN) Time: MO OC Test No.: 'IMla19 Test Volume: 25D O Scientific Name: f.@/3Wd**> Adults isolated . Dateh d4
) Test Article:
I dvM Test Duration: % br5 Lot No.: 5 " IM3 b Organisms Pulled (Time, Datel: MA l
Sample No. : (Wi- /OEJ0lG,(C80 Source: N Dilvent: drrh)or Acclimation: -
or Age 3 da ==
TARGET VALUES Culture Water TIS. 9 Transfer i* 2) M,Y Dilution No.: d Temp: JOlI DO: 'PI r+ ! I-pH: b~9 Salinity: @
ALL. TESTS Number of Conc. Rep. Live Organisms Dissolved Oxygen Conductivity fuS/cm)
Temperature (*Cl pH tmg/L) or % No -- ~. L, "i ; .. -
Other o 24 am 12 en o 2a am 72 na n na 24 da 72 o 24 da 72 nn 0 24 da 72 en a 24 da 72 as OeM A to 9 9 7 9 m EiiW.fN3 h// f.: t) 7.7 7/f G f.8 99 4 f4 gf M A,4 a 27 pr a io g ,
& v o a.
.o h"
us-Itd ) It*/ R to 16 to /0 fo MM nt W W.4 7,9 9.l 'g .i D KJ g.5 1b g.g 9. c 7) gcg .1gr 3032999 (b to to l0 (0 10
- g3.((fM) 100 l. O to 3 Ik / /0 N M M.'1 M7 Pl 1.i 73 t.1 f.0 D V.7 66 Ws 1.4 77 %1 at 6
'551 3Y9 h
- is to to ID to
' is-ide ) Icel = tt3 to f0 .91 n.f ao.o #7 kl tt is 5O7 pW A ID P>
S.1 9./ Q P4 % si x3 3e3 16 to es 10 (0
. .s '
Instrument Number 7h h 3 J h M 7h Ik % l g % [{ T( h 9h %7 h Co.,ecton racio, in i.4 t.r +5 .Y K t to % V, % nc r.a p r4 6 k 7.4, %
Time 19 6 f2C # G )F Fth tv # E35 ile 4 iM W lgs nul N22 so nn 195 if 4 # #e f0f Investigator C[- Ch $ f (O @ Cl4 # f @ % Ct% 104- F%
ao f (d #" [ $ f ctt f f NI
- t. 6 la D .
AT9 AM
' I
) AQUATIC TOXICOLOGY BENCH SHEET SET UP (SMALL VOLUMES) i Project No.. 9 o o o 3 .i o Client- M '"
, QC Test No.- Nd 4
2
- Date/ Time Activity investigator )
Dilutions Made .
K i
/o- M r pao d Test Vessels Fille I
1 1 l
/f#O Transferred Organisms CAJ S
/9I Head Counts d //-
t 1
l 4 Sample No.- N' ION < /06 MO Dilution No.: h kI W l 1
Volume Test t J Test Concentration Material (ml) Final Volume l
, &h'h I 0 vJ- SDCL s
TC IN)
I 00 l. sQD D rL -
. l j& -ivy l00 .
kJ }K - /&)o l 00 1. an.m.
2127195 l
Chain-of-Custody R: cort EA EngiIeerirg, Scir c0, .
cnd Tc:hnlogy FOR OFFICE USE ONLY EA AquateToxico6ogy Laboratones y,,AQ, g,m\*, Species to be tested:
15 LovetonCarcie Sparks (Balbrnore). Maryland 21152 D. manga Mer%a sp D. pulex P.pugo Tclophone:(410)T71-4950 9 C. dubia C. vanegatus Ftx(41 771-4204 P. promelas Chent M ProjectNo.: Other M. batua Other A = Acute C = Chomic ca a lE~~,1,W B = Beaccumulatoi NPDES Number- ..
Order r: ,
StawCaryccmiyConcied-PLEASEREAD SAMPUNG INSTRUCTIONS ON BACK0FFORM Accession j Collection Sample Desenption Number i Start End (ndudingSite, Station NumberNolume loffice use only) Grab Composite Datemrne Datemme Number,andOutfallNumber) ofContamer J)pf-$09$l W hs Y -_ f $ 5 b h N lf Y W- lDf1 l l $$ '
l mr ,& Gt"& 'TY:_ G :n
- c er a ^ )> w
/
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! I i
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i Sam led By: Datemme Received By: Datemme
. Q Sampler's printed Name:
-~
/OfY W
Title:
RelinquishedBy: Datemme
\ t Refin utshedB Datamme By
'Rpivec
, J mVW/8W 2CWlhG ' Datemme i dr orn Sample Chilled Dunng Collection?ho Sample Shippe y;(circle)
Comments: Fed.Ex. Puro. UPS Aarbome (g\v N Cpm e-up.A e W J Other
,w sw v a4 3. e ra.ced .,h l AirBAlNumber.
Sample Temperature at Receipt: C
= =- v. -.,_o,, , ~ , . . . . .
s ,
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! I J l 4
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i !
4 i ATTACHMENT III
- i REPORT QUALITY ASSURANCE RECORD E
.I 1
4 2
i i
3 i
A 4
4 4
l J
. - . . - . - _ . . - - - . - - . . - - . - _ . - - - - . . - ~ _ _ - - - . - - - - - - -
i REPORT QUALITY ASSURANCE RECORD Client: Sin- Nen +i n EA Report No.: ALVI 1 0 ~ I A
! Project Number: 70on2 80 Type Analysis: Heo4e Author: rania O beLn Tat Organisms: 3. man nn f b f MMEloS L
REPORT CHECKLIST ,
1
), GA/QC M REVIEWER D&IE
- 1. Samples collected, transported, and received g f according to study pian requirements. un % > r. - -
- b. Ad g / o /2 (/9 F
- 1
. g
- 2. Samples prepared and processed according to l
j study pian requirements. - - , > r, O _lm /0 / 2 Y /9
- i
- 3. Data collected using calibrated equipment. 7 n> i n . ad ~ fo /2 Y / 9 S' g
i i 4. Calcuistions checked:
- Hand calculations checked M a e e > . r. n 1.Jo l -
Documented and verified statistical j procedure used. D o y 1. a <, AJ t i
j 5. Data input / statistical analyses complete and
- .:= ~~~- wiuk, ;d,n,
2 " C ."*m"'" *t Ylaf+aff.rd Iehr/rr
,ian O"!" *""*"" ** **"'" d O. k, to /2s/9s-U i l
I Aunion DAIE
- 9. Commentary reviewed and resolved. >*a>-a O, d /o/30/9f io. = study ,i.o and ouaiity as.ur. ,ceicont,oi Manve seen , net and tse ,e,o,t is approved:
W i (2. } & ioho19s-
' DAlt l
PROJIfT MANAGER Lh k &FyCEV ALITY TP Is/ der
' DXTE Uc 'A to/3olar SENIORMCHNIDAL REVIEWER DATE i ATS-QA12 3/10/93