## BIOMONITORING REPORT

FOR 

# NACHURS ALPINE SOLUTIONS INDUSTRIAL ALPINE ICE
- MELT DEICER

LAB ID # 0823019

JULY 2008

PREPARED BY:

ANALYTICAL LABORATORIES, INC. 1804 N. 33RD STREET BOISE, ID 83703

(208) 342
- 5515

### SUMMARY OF ANALYSES FOR CARGIL CENTRAL RESEARCH LAB # 0823019

The results for the Fathead Minnow survival study:

NOEC: 1.00g/L LOEC: 3.00 g/L IC50: 1.92 g/L

The resu lts for the Fathead Minnow growth study:

NOEC: 1.00g/L LOEC: 3.00 g/L IC50: 0.46 g/L

The results for the Ceriodaphnia dubia reproduction study:

NOEC: <0.03 g/L LOEC: 0.03 g/L IC50: 0.37 g/L

The results for the Ceriodaphnia dubia survival study:

NOEC: 0.25 g/L LOEC: 0.50 g/L IC50: 0.38 g/L

The results for the Selenastrum growth study:

NOEC: 3.00 g/L LOEC: >3.00 g/L IC50: >3.00 g/L

Chronic Toxicity was found in the freshwater flea (Ceriodaphnia dubia) reproduction test at all o f the concentrations tried and in the survival test at concentrations of .50, 1.0 and 3.0 g/L. Chronic Toxicity was also found in both the Fathead Minnow survival and growth tests at concentrations of 3.0g/L.

IntroductionToxicity analyses, con sisting of three chronic bioassays, EPA Test Method 1000.0, EPA Test

Method 1002.0 and EPA Test Method 1003.0 were conducted on deicer sam ples submitted by NACHURS
ALPINE SLOUTIONS INDUSTRIAL. Samples were sent to Analytical Laboratories, Inc. for analyse s. Toxicity Method 1003.0 utilizing the green algae Selenastrum capricornutum was initiated on July 25 and
completed on July 28, 2008. Toxicity Method 1000.0 utilizing the freshwater fathead minnow Pimephales
promelas larvae and toxicity method 1002.0 util izing the cladoceran Ceriodaphnia dubia were conducted

simultane ously from July 25 to August 1, 2008. Testing was conducted according to Methods for
Measuring the Chronic Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms,

Fift h Edition, EPA

- 821

- R

- 02

- 013; and Standard Methods for the Examination of Water and Wastewater,

19 th Ed i tion.

Methods and Materials
Test methods are designed to estimate and measure chronic toxicity of whole and partial effluents

to the cladoceran Ceriod aphnia dubia and the fathead minnow Pimephales promelas and in a 7

- day static
renewal test. The green algae Selenastrum capricornutum was exposed in a static system to a series of

concentrations of effluent for 96 hours. Effluent was used, whole or combine d, with artificially prepared
dilution water to prepare dilution series. Di lution water was prepared (eight parts Millipore Milli

- Q water
to two parts Perrier Mineral Water) to produce a moderately hard dilution and control water. Water was
prepared in b ulk 24 hours prior to analyses and was aerated co n tinuously to produce an initial pH of 7.9 to

8.3.
For Test Method 1002.0, adult female daphnias were selected from a supplied stock prior to
analysis start. Each individual was inspected for proper behav ior (swim pattern, physical ap pear ance)
before placement in culture dishes. Neo nates 4

- 6 hours old were selected randomly from a com posite pool,
inspected, and arranged in six dil u tions in cluding control. Analyses at a static renewal were performed

o ver the next seven co n secutive days. Data obtained was used to de termine both NOEC and LOEC for

survival and re production (see Appendix I

- Definition of Terms.)

Test Design/Standard Conditions Method 1002.01. Test Type

- static renewal (d aily)

Collection #1

- July 25, 2008

Day 7

- Review

2. Temperature

- 25 degrees C. + or

- 1 degree C.

3. Light Quality

- incubator chamber (Environmental Chamber Fisher/11

- 67966)

4. Light Intensity

- incubation chamber (as above)

5. Photoperiod

- 16 hrs. light; 8 hrs. dark

6. Test Chamber

- 30 ml. anchor

- hocking

7. Renewal

- all dilutions daily

8. Age (neonates/less than 24 hours

— typically 4

- 6 hours)

9. Organisms per chamber

- one

10. Replicates

- ten chambers/control and each dilution

11. Feeding

- 0.1 ml. YCT; 0.1 ml. Selenastrum suspens ion

- daily

12. Dilution water

- 20% diluted mineral water (reference section)

13. Concentrations used

– .03 g/L, .25 g/L, .5 g/L, 1.0 g/L, 3.0 g/L and control

14. Duration

- 60% of controls have had 3 broods and/or at least 7 days

15. Endpoint

- survival/reproduction

16. Acc eptability

- 80% or greater of control survival/60% of con trol produce 3 rd brood; average young
production 15 or greater per surviving female

17. Source of organisms

– Aquatic Biosystems, Fort Collins, Colorado

For Test Method 1000.0, utilizing fathead minn ow Pimephales pro melas , larvae were sent from
Aquatic Biosystems in Fort Collins, Colorado. Organisms were sent by Federal Express in oxy gen saturated water contained in plastic bags. Once received, larvae were transferred to lab o ra tory control water

an d allowed to acclimate for eight hours prior to transfer to test solution. At transfer, larvae were
individually inspected under dissection m i croscope for deformities and ab normal behavior. Healthy larvae
were transferred to test cells using wide

- bore pip ettes. Larvae were offered freshly hatched, freshwater

rinsed brine shrimp artemia nauplii . Larvae were fed and water renewed daily using fresh test solution for
seven consecutive days. Data obtained was used to determine NOEC and LOEC for survival and g rowth
(weight gain.)

Test Designed/Standard Conditions/Method 1000.0Test design and standard conditions for Method 1000.0 are as fo l lows:

1. Test Type

- static renewal (daily)

Collection #1

- July 25, 2008

Day 7

- Review
2. Temperature

– 25 deg rees C. + or

– 1 degree C.
3. Light Quality

– incubator chamber (Environmental Chamber Fisher/11

- 67966)
4. Light Intensity

– incubation chamber (as above)
5. Photoperiod

- 16 hrs. light; 8 hrs. dark

6. Test Chamber

- 500 mL borosylicate glass beakers

7. Test Solution Volume

- 250 ml./replicate

8. Renewal static

- all dilutions daily
9. Age of Test Organisms

- newly hatched, less than 36

- hours old

10. Individual/Chamber

- 10 per chamber
11. Chamber Replicates

- 4 replicates of control and each dilution
12. Feeding

- 0.1 ml. newly hatched brine shrimp nauplii two times daily, 8 hr. intervals

13. Dilution Water

– 20% diluted mineral water (reference Section 7)

14. Dilution Concentrations

– .03 g/L, .25 g/L, .5 g/L, 1.0, 3.0 g/L and co n trol

15. Test Durati on

- 7 days
16. Endpoints

- Survival and growth (individual dry weight)
17. Acceptability

- 80 % survival in controls. Dry weight of sur viving controls equals or exceeds 0.25
mg/individual
18. Sample Volume Taken

- 1 gallon for test solution renewal/1 l iter for daily composite water chem istries
19. Source of Organisms

- Aquatic Biosystems, Fort Collins, ColoradoFor Test Method 1003.0, utilizing the green algae Selenastrum capricornutum , starter cultures are
initiated by aseptically transferr ing 1 mL to a culture flask containing algal culture medium 4 to 7 days
prior to test initiation. Inoculum was prepared no more than 2 to 3 hours prior to the beginning of the test.
Initial inoculum was estimated to contain 1,000,000 cells per mL added to 100 mL dilution yielding a final
cell concentration of approximately 100,000 per dilution replicate. Vessels were shaken twice daily for the
duration of the test period to avoid settling. Data obtained was used to determine NOEC and LOEC for

increase in cell density.

Test Designed/Standard Conditions/Method 1003.0
Test design and standard conditions for Method 1003.0 are as fo l lows:

1. Test Type

- static system

Collection #1

– July 25, 2008

96 hour

- Review
2. Temperature

- 25 degrees C. + or

- 1 degree C.
3. Light Quality

– incubator chamber (Environmental Chamber Fisher/11

- 67966)
4. Light Intensity

– incubation chamber (as above)
5. Photoperiod

– 24 hrs light

6. Test Chamber

– 250 mL borosylicate glass bottles

7. Test Solution Volume

– 10 0 ml./replicate
8. Age of Test Organisms

– 4 day culture.

9. Individual/Vessel

– 100,000 per mL
10. Vessel Replicates

– 4 replicates of control and each dilution
11. Feeding

– Initial addition of Algal culture medium at equal portion to dillution.

12. Di lution Water

- 20% diluted mineral water (reference Section 7)

13. Dilution Concentrations

– .03 g/L, .25 g/L, .5 g/L, 1.0g/L, 3.0 g/L and co n trol

14. Test Duration

– 96 hours
15. Endpoints

– Measurement of cell density using a Sedgwick

- Rafter hemocytomet er . 16. Acceptability

– cell density in the control flasks must exceed 1 X 10 5 and not vary more than 20%
among replicates
17. Sample Volume Taken

– approximately 500 ml for test dilutions of, .03 g/L, .25 g/L, .50 g/L, 1.0g/L,
3.0g/L and control for c hemistries.

18. Source of Organisms

- Aquatic Biosystems, Fort Collins, Colorado

Water chemistries performed on received effluent samples con sisted of Chlorine Residual, Alka

linity, Conductivity, Hardness, Ammonia, and pH (see water chemistries and test dat a at tached as Table IV;

Raw data is attached as A p pendix VI and VII.)Interpretation

- Statistical Review
Survival data for Method 1002.0 was first analyzed after tabula tion using Fishers Exact test. This
test individually determines statistical signi ficance between control and effluent dilu tions at tempted. Once
survival significance was analyzed, the NOEC and LOEC for survival were inde pendently determined.
Reproduction data was analyzed first by Chi

- Square or Shapiro Wilk’s test for normality an d then by

Bartletts or Levene’s test for variance. Once normal ity and type of variance was established, the data was
formally tested by the hypothesis approach (Dunnett’s or Bonferroni t

- Test) to determine NOEC and LOEC
for reproduction. This same appro ach was applied to both survival and growth data for Methods 1000.0

and 1003.0.

Results

- Method 1002.0
During EPA Method 1002.0, Survival and Reproduction test using Ceriodaphnia dubia , survival
values from specific dilutions of collected deicer were measured and com pared to val ues obtained from
control individuals set up in 20% diluted mineral water. For survival, concentrations of .50, 1.0 and 3.0g/l
clearly exhibit a significant difference from controls. Statisti cal analyses of data also indicat e that these
dilutions are significantly different from controls.
For reproduction, all of the concentrations failed to meet the minimum reproduction rate of 15
organisms per adult in 7 days or three broods. Statistical analyses of reproduction also indica te that all
concentrations are statistically different from controls.

Endpoints Determined

- EPA Method 1002.0

NOEC LOEC
Ceriodaphnia dubia Survival 0.25 g/L 0.50 g/L
Reproduction <0.03 g/L 0.03 g/L

The mortality was less than twenty percent (<20%) in controls. Test was declared valid.Results

- Method 1000.0
During EPA Method 1000.0, larval Survival and Growth test using the fathead minnow Pimephales
promelas , survival and growth from specific diluti ons of deicer wastewater were measured and com pared to
values obtained from controls prepared in 20% diluted mineral water. Statisti cal analyses of survival data
indicate that the dilution of 3.0g/L presents a significant difference from controls. Growth , as a measure of
total dry weight, also shows a significant difference from controls exists in the 3.0g/L dilution. Statistical
analyses of growth data indicated that concentrations of .25, .50, 1.0 and 3.0g/L are significantly different

from controls. O f theses concentrations, .25, .50 and 1.0g/L successfully met the minimum weight

- gain
requirement of 0.25mg per individual and are therefore declared passing.

Endpoints Determined

- Method 1000.0

NOEC LOEC
Pimephales promelas Survival 1.00 g/L 3.00 g/L

Growth 1.00g/L 3.00 g/L

Survival of controls exceeded eighty percent (80%). Net dry weight of surviving indi viduals ex

ceeded 0.25 mg/individuals in all concentrations except one. Test was declared valid.

Results

- Method 1 003.0
During EPA Method 1003.0, Selenastrum growth response test using the green algae Selenastrum
capricornutum Growth from specific dilutions of collected deicer were measured and com pared to values
obtained from controls prepared in 20% diluted minera l water. Statistical analyses of growth data indicate
that none of the concentrations tried are significantly different from controls.

Endpoints Determined

- Method 1003.0

NOEC LOEC
Selenastrum capricornutum Growth 3.0 g/L >3.0 g/L
Survival of controls exceeded 100,000 cell density. Test was declared valid.Test Quality Control
Quality control practices for effluent toxicity tests include certain precautions at each of the fo l

lowing steps:

1. Effluent sampling and handl ing . Sampling containers prepared as per section 7 of Methods for
Measuring and Chronic Toxicity of Effluent to Freshwater and Marine Organisms were pro vided to
client. Transportation containers containing sample and chain of custody were sent to Analyt ical Labs
for analysis.

2. Condition of test organisms . Organisms for Method 1002.0 are separated from stock culture two
weeks prior to test start

- up. Individual survival and reproduction rates are monitored and recorded.
Test organisms are selected random ly from the off

- spring of these monitored indi viduals, thus
minimizing any possible stress prior to testing and to assure a stable, healthy, and similar aged test
individuals. Test organisms for EPA Method 1000.0 are purchased from Aquatic Biosystems, Fo rt
Collins, Colorado, a state and federally approved aquatic test o r ganism supplier. (See Appendix VIII.)
Test organisms for EPA Method 1003.0 were cultured 4 to 7 days prior to test initiation.

3. Conditions of test equipment . All test equipment at a min imum is maintained according to listed
references or manufacturer’s specifications. Equipment such as balances, thermometers, and
spectrophotometers are calibrated annually by outside sources and certificates are main tained. All
equipment use and mai n t enance records, along with calibrations made, are re corded in a mainte nance log for that sp e cific equipment.

4. Test conditions . Only test methods directly from EPA refer ences or methodologies provided by EPA
are used. Any deviations or alterations from these procedures are documented and approved prior to

use.

5. Reference toxicants . Reference toxicants are used for both EPA Methods 1000.0 and 1002.0. Sodium
chloride is made up in dilution control water at prescribed concentrations and is used to determin e Test

LC50 for each method. Reference toxi cants are run concurrently with each proce dure and are plotted
over time. (See Appendix IX.)

6. Record Keeping . All raw data along with data evaluation is included in report to client. Original data
along with all test records are maintained at laboratory for client or future re f er ence.LIST OF TABLES AND APPENDICES

Table I

- Ceriodaphnia dubia Survival and Reproduction Data

- Method 1002.0

Table II

- Pimephales promelas Survival Data

- Method 1 000.0

Table III

- Pimephales promelas Growth Data

- Method 1000.0

Table IV

- Water Chemistries

- Daily Renewal Summary

Table V

- Selenastrum capricorntum water quality and temperature

- 1003.0

Table VI

- Selenastrum capricornutum cell density summary

- Me thod 1003.0

Appendix I

- Definition of Terms

Appendix II

- Ceriodaphnia dubia Raw Data

Appendix III

- Pimephales promelas Raw Data

Appendix IV

- Selenastrum capricorntum Raw Data

Appendix V

- Deicer chemistry reports

Appendix VI

- Organisms (fish)

- Transfer Sheets

Appendix VII

- Literature Cited

Appendix VIII

- Reference Toxicants

## TABLE I

CERIODAPHNIA BIOASSAY

– SUMMARY

METHOD 1002.0

NACHURS ALPINE SOLUTIONS INDUSTRIAL

ALPINE ICE

- MELT

JULY 2008

EFFLUENT PERCENT INITIAL COUNT 48

- HOUR COUNT 96

- HOUR COUNT FINAL COUNT PERCENT SURVIVAL AVERAGE YOUNG/
FEMALE

Control 10 10 10 10 100 18.7 0.03 g/L 10 10 10 09 90 12.4 0.25 g/L 10 10 10 09 90 12.1 0.50 g/L 10 10 10 09 90 6.3 1.00 g/L 10 10 00 00 00 0.0 3.00 g/L 10 00 00 00 00 0.0

## TABLE II

FAT HEAD MINNOW LARVAE

- SURVIVAL SUMMARY

NACHURS ALPINE SOLUTIONS INDUSTRIAL

ALPINE ICE

- MELT

METHOD 1000.0

JULY 2008

EFFLUENT PERCENT INITIAL COUNT 24

- HOUR COUNT 48

- HOUR COUNT 96

- HOUR COUNT FINAL COUNT PERCENT
SURVIVAL

Control 40 40 40 40 37 93 0.03 g/L 4 0 40 40 39 36 90 0.25 g/L 40 40 40 38 37 93 0.50 g/L 40 40 40 39 31 78 1.00 g/L 40 40 40 39 36 90 3.00 g/L 40 29 22 04 01 2.5

## TABLE III

FATHEAD MINNOW LARVAE

- GROWTH SUMMARY

NACHURS ALPINE SOLUTIONS INDUSTRIAL

ALPINE ICE

- MELT

JULY 2008

EFFLUENT PERCENT APPROXIMATE AVERAGE INITIAL

WEIGHT (MG)* ENDING AVERAGE WEIGHT (MG) ENDING AVERAGE WEIGHT GAIN (MG)

Control 0.10 0.96 0.86 0.03 g/L 0.10 0.80 0.70 0.25 g/L 0.10 0.60 0.50 0.50 g/L 0.10 0.46 0.36 1.00 g/L 0.10 0.43 0.3 3 3.00 g/L 0.10 0.008

- 0.09 * Initial weight obtained by taking 40 individuals at beginning of procedure (weight is dry weight/mg, 105 ° C. for 24 hours.) Net weight total used for growth calculation, not weight gain.

## TABLE IV

WATER CHEMISTRIES , DAILY RENEWALS

- pH & DISSOLVED OXYGEN

METHOD 1002.0

– NACHURS ALPINE SOLUTIONS

INDUSTRIAL

ALPINE ICE

- MELT

JULY 2008

Day #0 #1 #2 #3 #4 #5 #6 #7

Control

- pH DO 7.8 7.2 7.2
7.5 7.0 7.6 7.6 7.0 8.1 7.4 8.1
7.5 8.5 7.2 8.0 7.6 .03 g/L

- pH DO 7.5 6.9 8.1
7.7 7.7 8.0 7.3 7.3 8.4 7.4 8.0
7.7 8.4 6.6 8.0
7.7 .25 g/L

- pH DO 7.5 6.9 8.2 7.6 8.2 7.1 8.0 6.2 8.2 6.7 8.0 7.0 8.1 7.0 7.8 7.1 .50 g/L

- pH DO 7.4 7.0 8.3 6.8 8.1 6.8 8.0 6.1 8.2 6.6 7.2 7.2 7.5 6.4 7.5 6.8 1.0 g/L

- pH DO 7.7 7.0 8.4 6.8 8.1 6.6 3.0 g /L

- Ph DO 7.8 7.2 8.4 6.9

## TABLE IV

WATER CHEMISTRIES, DAILY RENEWALS

- pH & DISSOLVED OXYGEN

METHOD 1000.0

– NACHURS ALPINE SOLUTIONS

INDUSTRIAL

ALPINE ICE

- MELT

JULY 2008

Day #0 #1 #2 #3 #4 #5 #6 #7

Control

- pH DO 7.2 7.8 7.9 5.4 7.4 4.9 6.7 3.9 7.6 4.5 6.8 3.0 7.7 3.7 9.7 3.0 .03 g/L

- pH DO 7.5 6.9 7.8 4.6 7.4 4.6 7.1 3.8 7.4 4.1 7.2 3.1 7.6 3.1 7.9 3.3 .25 g/L

- pH DO 7.5 6.9 7.8 4.9 7.5 3 .3 7.2 1.0 7.7 2.8 7.2 2.1 7.6 3.8 7.9 2.8 .5 g/L

- pH DO 7.4 7.0 7.9 3.9 7.6 3.3 7.4 1.3 7.8 7.6 7.3 1.9 7.8 2.9 8.0 1.4 1.0 g/L

- pH DO 7.7 7.0 7.9 3.5 7.5 3.3 7.4 1.2 6.9 2.8 7.6 1.8 7.9 2.1 7.9 1.4

3.0 g/L

- pH DO 7.8 7.2 7.9 3.6 7.8 2.9 6.9 1.0 7.7 1.4 1.0 7.0 7.1 1.2 7.1 1.0

## TABLE V

WATER CHEMISTRIES, DAILY RENEWALS

- pH & DISSOLVED OXYGEN

METHOD 1003.0

– NACHURS ALPINE SOLUTIONS

INDUSTRIAL

ALPINE ICE

- MELT

JULY 2008

EFFLUENT CONC. CONTROL .03 g/L .5 g/L 3.0 g/L pH

Alkalinity

Hardness

Conductivity

Chlorine

Ammonia 7.8

71.1 mg/L

82.5 mg/L

171 umhos

<0.10 mg/L

<0.04mg/L 7.8

73.3 mg/L

80.3 mg/L

196 umhos

<0.10 mg/L

<0.04 mg/L 7.9

130 mg/L

78.8 mg/L

442 umhos

<0.10 mg/L

<0.04mg/L 7.8

424 mg/L

80.5 mg/L

1,680 umhos

<0.10 mg/L

<0.04

## TABLE VI

SELENASTRUM CAPRICORNUTUM GROWTH RESPONSE

SUMMARY

METHOD 1003.0

– NACHURS ALPINE SOLUTIONS

INDUSTRIAL

ALPINE ICE

- MEL T

JULY 2008

EFFLUENT PERCENT INITIAL CELL DENSITY 96

- HOUR CELL DENSITY REPLICATE 1 96

- HOUR CELL DENSITY REPLICATE 2 96

- HOUR CELL DENSITY REPLICATE 3 96

- HOUR CELL DENSITY REPLICATE 4 MEAN GROWTH RESPONSE (Y I 

### )

Control

Log 10 100,000 200,000 5.30 205,000 5.31 130,000 5.11 181,000 5.26 179,000 5.25 .03 g/L Log 10 100,000 156,000 5.19 200,000 5.30 163,000 5.21 177,000 5.25 174,000 5.24 .25 g/L Log 10 100,000 150,000 5.18 230,000 5.36 158,000 5.20 162,000 5.21 175,000 5.24 .5 g/L Log 10 100,000 185,000 5.27 1 79,000 5.25 161,000 5.21 172,000 5.24 174,000 5.24 1.0 g/L Log 10 100,000 139,000 5.14 142,000 5.15 178,000 5.25 212,000 5.33 168,000 5.23 3.0 g/L Log 10 100,000 152,000 5.18 215,000 5.33 213,000 5.33 172,000 5.24 188,000 5.27Appendix I



### Terms*

1. Safe Concentration . The highest concentrations of toxicant that will permit normal

propaga tion of fish and other aquatic life in receiving waters, biologically defined

rather than statist i cally.

2. N.O.E.C. (No

- Observed Effect Concentration)

- The highest concentration of
toxicant to which organisms are exposed in a full

- life cycle or partial

- life cycle test

that causes no o b serv able, adverse effects on the test organisms (the highest
concentration of toxicant in which the values for the observed paramete rs are not
statistically, significantly different from controls), statistically d e fined.

3. L.O.E.C. (Lowest

- Observed Effect Concentration)

- The lowest concentration of
toxicant to which organisms are exposed in a full

- life cycle or a partial

- life cycle test

which causes ad verse effects on the test organisms (where values for the observed

parameters are statistically, significantly different from the co n trols).

4. TUc (chronic toxicity units)

– HIGHEST CONCENTRATION TRIED/NOEC

5. IC50 (Inhibition Concentration 50 %)

– A 50% reduction in the reproduction or
growth of the test organisms.

*Taken from “ Short

- Term Methods for Estimating the Chronic *Toxicity of Effluent and
Receiving Waters to Freshwater Organisms, 3 rd 

### Edition, EPA/600/4

- 91/002App endix VI

Literature Cited

1. Short

- Term Methods for Estimating the Chronic Toxicity of Effluent and Receiving Waters to
Freshwater Organisms , 3 nd Ed i tion, EPA/600/4

- 91/002.

2. Methods for Measuring the Chronic Toxicity of Effluents to Freshwater and Marine Orga nisms

, EPA/600/4

- 85/013, US EPA.

3. Standard Methods for the Examination of Water and Wastewater , 19 Edition, 1995, APHA, AWWA,

WPCF.
Handbook for Analytical Quality Control in Water and Wastew a ter Laboratories , Environ mental Monitoring and Support Labor a tor y, Cincinnati, EPA/600/4

- 79/019, US EPA

### Green Algae, Selenastrum capricornutum Growth Test Method 1003.0

Discharger: De

- Icer #0823019 Test Dates: 7/25/08

- 8/01/08

Location: NACHURS ALPINE SOLUTIONS IN DUSTRIAL

- ALPINE ICE

- MELT



## Day 1 Day 2 Day 3 Day 4

Control

- pH Temp (°C) Cell Density (x10 3 

### ) 7.8 23.4 179 8.1 24.5

-- 9.2 25.0

-- 9.5 24.1 217

0.03 g/L

- pH Temp (°C) Cell Density (x10 3 

### ) 7.9 23.4 174 7.8 24.5

-- 8.8 25.0

-- 9.3 24.1 250

0 .25 g/L

- pH Temp (°C) Cell Density (x10 3 

### ) 7.9 23.4 175 8.0 24.5

-- 8.7 25.0

-- 9.3 24.1 233

0.50 g/L

- pH Temp (°C) Cell Density (x10 3 

### ) 7.9 23.4 174 8.2 24.5

-- 8.8 25.0

-- 9.5 24.1 249

1.0 g/L

- pH Temp (°C) Cell Density (x10 3 

### ) 8.0 23.4 1 68 8.3 24.5

-- 8.9 25.0

-- 9.4 24.1 251

3.0 g/L

- pH Temp (°C) Cell Density (x10 3 

### ) 8.0 23.4 188 8.2 24.5

-- 8.5 25.0

-- 8.9 24.1 237