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 results 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 of 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. 

 

 

Introduction
Toxicity analyses, consisting of three chronic bioassays, EPA Test Method 1000.0, EPA Test 

Method 1002.0 and EPA Test Method 1003.0 were conducted on deicer samples submitted by NACHURS 

ALPINE SLOUTIONS INDUSTRIAL. Samples were sent to Analytical Laboratories, Inc. for analyses. 

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 utilizing the cladoceran Ceriodaphnia dubia were conducted 

simultaneously 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,

Fifth Edition, EPA-821-R-02-013; and Standard Methods for the Examination of Water and Wastewater,

19th Edition. 

Methods and Materials

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

to the cladoceran Ceriodaphnia 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 combined, with artificially prepared 

dilution water to prepare dilution series. Dilution 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 bulk 24 hours prior to analyses and was aerated continuously 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 behavior (swim pattern, physical appearance) 

before placement in culture dishes. Neonates 4-6 hours old were selected randomly from a composite pool, 

inspected, and arranged in six dilutions including control. Analyses at a static renewal were performed 

over the next seven consecutive days. Data obtained was used to determine both NOEC and LOEC for 

survival and reproduction (see Appendix I - Definition of Terms.)

Test Design/Standard Conditions Method 1002.0
1. Test Type - static renewal (daily)

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 suspension - 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. Acceptability - 80% or greater of control survival/60% of control produce 3rd 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 minnow Pimephales promelas, larvae were sent from 

Aquatic Biosystems in Fort Collins, Colorado. Organisms were sent by Federal Express in oxygen 

saturated water contained in plastic bags. Once received, larvae were transferred to laboratory control water 

and allowed to acclimate for eight hours prior to transfer to test solution. At transfer, larvae were 

individually inspected under dissection microscope for deformities and abnormal behavior. Healthy larvae 

were transferred to test cells using wide-bore pipettes. Larvae were offered freshly hatched, freshwaterrinsed 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 growth 

(weight gain.) 

Test Designed/Standard Conditions/Method 1000.0
Test design and standard conditions for Method 1000.0 are as follows:

1. Test Type - static renewal (daily)

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 - 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 control

15. Test Duration - 7 days

16. Endpoints - Survival and growth (individual dry weight)

17. Acceptability - 80 % survival in controls. Dry weight of surviving controls equals or exceeds 0.25 

mg/individual

18. Sample Volume Taken - 1 gallon for test solution renewal/1 liter for daily composite water chemistries

19. Source of Organisms - Aquatic Biosystems, Fort Collins, Colorado
For Test Method 1003.0, utilizing the green algae Selenastrum capricornutum, starter cultures are 

initiated by aseptically transferring 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 follows:

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 – 100 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. Dilution 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 control

14. Test Duration – 96 hours

15. Endpoints – Measurement of cell density using a Sedgwick- Rafter hemocytometer . 

16. Acceptability – cell density in the control flasks must exceed 1 X 105 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 chemistries.

18. Source of Organisms - Aquatic Biosystems, Fort Collins, Colorado

Water chemistries performed on received effluent samples consisted of Chlorine Residual, Alkalinity, Conductivity, Hardness, Ammonia, and pH (see water chemistries and test data attached as Table IV; 

Raw data is attached as Appendix VI and VII.)
Interpretation - Statistical Review

Survival data for Method 1002.0 was first analyzed after tabulation using Fishers Exact test. This 

test individually determines statistical significance between control and effluent dilutions attempted. Once 

survival significance was analyzed, the NOEC and LOEC for survival were independently determined. 

Reproduction data was analyzed first by Chi-Square or Shapiro Wilk’s test for normality and then by 

Bartletts or Levene’s test for variance. Once normality 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 approach 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 compared to values 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. Statistical analyses of data also indicate 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 indicate 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 dilutions of deicer wastewater were measured and compared to 

values obtained from controls prepared in 20% diluted mineral water. Statistical 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. Of 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 individuals exceeded 0.25 mg/individuals in all concentrations except one. Test was declared valid.

Results - Method 1003.0 

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 compared to values 

obtained from controls prepared in 20% diluted mineral 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 following steps:

1. Effluent sampling and handling. Sampling containers prepared as per section 7 of Methods for 

Measuring and Chronic Toxicity of Effluent to Freshwater and Marine Organisms were provided to 

client. Transportation containers containing sample and chain of custody were sent to Analytical 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 randomly from the off-spring of these monitored individuals, 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, Fort 

Collins, Colorado, a state and federally approved aquatic test organism 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 minimum 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 maintained. All 

equipment use and maintenance records, along with calibrations made, are recorded in a maintenance 

log for that specific equipment.

4. Test conditions. Only test methods directly from EPA references 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 determine Test 

LC50 for each method. Reference toxicants are run concurrently with each procedure 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 reference. 
LIST OF TABLES AND APPENDICES

Table I - Ceriodaphnia dubia Survival and Reproduction Data - Method 

1002.0

Table II - Pimephales promelas Survival Data - Method 1000.0

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-Method 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

FATHEAD 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 40 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 0.33

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-MELT

 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

(YI)

Control

Log10 

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

Log10

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

Log10

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

Log10

100,000 185,000

5.27

179,000

5.25

161,000

5.21

172,000

5.24

174,000

5.24

1.0 g/L

Log10

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

Log10

100,000 152,000

5.18

215,000

5.33

213,000

5.33

172,000

5.24

188,000

5.27
Appendix I

Terms*

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

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

rather than statistically.

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 observable, adverse effects on the test organisms (the highest 

concentration of toxicant in which the values for the observed parameters are not

statistically, significantly different from controls), statistically defined.

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 adverse effects on the test organisms (where values for the observed 

parameters are statistically, significantly different from the controls).

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, 3rd Edition, EPA/600/4-91/002 
Appendix VI

Literature Cited

1. Short-Term Methods for Estimating the Chronic Toxicity of Effluent and Receiving Waters to 

Freshwater Organisms, 3nd Edition, EPA/600/4-91/002.

2. Methods for Measuring the Chronic Toxicity of Effluents to Freshwater and Marine Organisms, 

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 Wastewater Laboratories, Environmental 

Monitoring and Support Laboratory, 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 INDUSTRIAL-ALPINE ICE-MELT 

Day 1 Day 2 Day 3 Day 4

Control -pH

Temp (°C)

Cell Density 

(x103)

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 

(x103)

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 

(x103)

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 

(x103)

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 

(x103)

8.0

23.4

168

8.3

24.5

--

8.9

25.0

--

9.4

24.1

251

3.0 g/L -

pH

Temp (°C)

Cell Density 

(x103)

8.0

23.4

188

8.2

24.5

--

8.5

25.0

--

8.9

24.1

237