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BLOCKENVIRONMENTAL SERVICES
2451 Estand Way
Pleasant Hill, CA 94523-3911 (925) 682-7200 FAX 686-0399
Aquatic Toxicity Testing Results for Soilworks, LLC Three 96-Hour Product LC-50 Bioassays
Prepared For:
Chad Falkenberg Soilworks, LLC
681 N. Monterey St., Ste 101
Gilbert, AZ 85233-3818
BES Sample #22382
Prepared By:
Block Environmental Services, Inc.
2451 Estand Way Pleasant Hill, CA 94523
October 6, 2005
David Block, Ph.D
Laboratory Director
Nanette Malan Lake
Laboratory Manager
Soilworks of Gilbert, Arizona requested Block Environmental Services (BES) to perform 96-Hr LC-50 aquatic bioassays, using rainbow trout (Oncorhynchus mykiss), fathead minnows (Pimephales promelas) and the mysid (Americamysis bahia) for the product Durasoil™, a synthetic organic dust control agent.
BES received the product on July 25, 2005 and performed the definitive toxicity tests in order to determine concentration-response curves for fish mortality, LC-S0’s and the 95 percent confidence inte rvals.
A definitive test was performed for each species in order to estimate a concentration range within which the LC-50 value would fall. The definitive tests consisted of 5 concentrations and a laboratory control, run in duplicate.
The test protocols and guidelines followed the EPA Ecological Effects Test Guidelines (OPPTS 850.1075 Fish Acute Toxicity Test, Freshwater and Marine).
As a QNQC measure, standard reference toxicant tests were run to confirm good organism quality .
This report describes the procedures used and the results obtained for the aquatic toxicity tests initiated on July 27, 2005 and completed August 20, 2005.
BES is an Environmental Laboratory Accreditation Program certified laboratory (#1812).
BES# 22382 Page 2 of 8
2.1 Sample and Test Preparation
Sample identification – Durasoil™
Chemical name: Synthetic Organic Dust Control Agent Appearance and odor: Colorless, odorless
Solubility: Insoluble in water
The sample was received on July 25, 2005 and kept at ambient room temperature for the duration of the study.
Samples to set up the concentration series for the range finding tests were weighed out using a Sartorius Balance, disposable dispenser pipettes and clean glassware. The concentration series for the range finding tests are summarized in Table 1.
A summary of testing parameters by organism can be found in Table 2.
2.2 Organism information
Species: Rainbow trout (Oncorhynchus myldss)
Supplier: In house culture Stock date: 07/05/2005
Age: Juvenile(< 3.0g)
Species: Fathead minnow (Pimephales promelas)
Supplier: Commercial supplier Stock date: 07/20/2005
Age:
Species:
Juvenile(< 3.0g)
Mysid (Americamysis bahia)
Supplier : Commercial supplier Stock date: 08/16/2005
Age: 4 days old
BES# 22382 Page 3 of 8
2.3 Reference Toxicant
Sodium Dodecyl Sulfate was used as the reference toxicant for the rainbow trout and fathead minnows. . Potassium chromate was used as the reference toxicant for the mysid. The toxicity endpoints from the reference toxicant tests are subsequently plotted on a running control chart from the last 20 tests. The mean values as well as the upper and lower control limits(± 2 standard deviations) are recalculated with each successive test result. The outliers, which are values falling outside the upper and lower control limits, and trends of increasing or decreasing sensitivity, are readily identified.
3.1 Test Procedures
A detailed procedure for each test is outlined in laboratory standard operating procedures (SOP’s) kept at the BES laboratory and are based upon the following references:
Additional guidelines used for these tests:
Test).
BES# 22382 Page 4 of8
3.2 Data Analysis
Toxicity testing results were analyzed using the software program ToxCalc Version 5.
The trimmed Spearman Karber method was used to determine the LC/EC values. The testing compared the sample responses with the laboratory control water. The parameters of interest are the LC-50 value and the 95% upper and lower confidence limits.
Table 1: Concentration series for the range finding tests (mg/L).
Organism |
||
0. mykiss |
P. promelas |
A. bahia |
Control |
Control |
Control |
1 |
1 |
1 |
10 |
10 |
10 |
100 |
100 |
100 |
1,000 |
1,000 |
1,000 |
10,000 |
10,000 |
10,000 |
BES# 22382 Page 5 of8
Table 2: Summary of Testing Parameters II
Test Conditions |
Organism |
||
0. mykiss |
P. promelas |
A. bahia |
|
Temperature |
12 ± 1 °C |
20 ± 1 °C |
25 ± 1 °C |
Light Intensity |
50-100 ft-c |
||
Photoperiod |
16hr light: 8hr dark |
||
Test Chamber |
6 L glass aquarium |
6 L glass aquarium |
400 ml glass beaker |
Test Solution Volume |
SL |
SL |
200ml |
Organism Age |
Juvenile(< 3.0g) |
Juvenile (< 3.Og) |
1-5 days |
Organism source |
Trout Lodge Inc., Sumner WA |
Thomas Fish Co., Anderson CA |
Aquatic Biosystems, Fort Collins, CO |
Organisms/chamber |
10 |
||
Chamber/ concentration |
2 |
||
Organisms/concentration |
20 |
||
Food Source |
None |
None |
Artemia nauplii |
Feeding Frequency |
None |
None |
Day O and at 48 hr |
Cleaning |
None |
||
Aeration |
None, unless D.O < 6.0 mg/L |
None, unless D.O < 4.0 mg/L |
None, unless D.O < 4.0 mg/L |
Reference Toxicant |
SDS |
SDS |
K2CrO4 |
Reference toxicant concentrations |
8, 16, 24, 32 and 40 mg/L |
8, 16, 24, 32 and 40 mg/L |
1.05, 2.1, 4.2, 8.5 and 16.9 mg/L |
Test Duration |
96 hr |
||
Effects Measured |
Survival |
||
Test Acceptability |
2:90% survival in control |
BES# 22382 Page 6 of 8
5.1 Rainbow trout (Oncorhynchus mykiss) results
Table 3 Range Fm’ d”mg Test Resu ts
Concentration (mg/L) |
Survival (%) |
Cl |
100 |
10 |
100 |
50 |
100 |
100 |
100 |
1,000 |
100 |
10,000 |
100 |
5.2
|
Fathead minnow (Pimephales promelas) results Table 4: Range Finding Test Results
5.3 Mysid (Americamysis bahia) results
Table 5:
Range Fma· mg Test Resu ts
Concentration (me/L) |
Survival (%) |
Cl |
100 |
10 |
100 |
50 |
100 |
100 |
100 |
1,000 |
100 |
10,000 |
100 |
5.4 Testing Notes
The Laboratory control water passed the acceptability criteria for survival (2:_90%) for the rainbow trout, fathead minnows and mysid 96 hr bioassays. The reference toxicant test results generated survival and growth endpoints that were within control chart limit ations. These responses indicate that the values elicited for the Durasoil™ product are valid.
BES# 22382 Page 7 of8
The objective of these tests was to determine the toxicological effects (if any) of the product, Durasoil™ on aquatic organisms that represent different thropic levels in the environment. The results for the definitive bioassay tests using the product Durasoil™, received in July 2005 indicate that:
BES# 22382 Page 8 of 8
Copyright Soilworks, LLC 2003-. All Rights Reserved. Soilworks®, Soiltac®, Gorilla-Snot®, and Durasoil®are registered trademarks of Soilworks, LCC.
Copyright Soilworks, LLC 2003-. All Rights Reserved. Soilworks®, Soiltac®, Gorilla-Snot®, and Durasoil® are registered trademarks of Soilworks, LCC.