Toxicity Assessment of Triple Strike. VGT Pty Ltd. Test Report

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1 Toxicity Assessment of Triple Strike VGT Pty Ltd Test Report January 2011

2 Toxicity Assessment of Triple Strike VGT Pty Ltd Test Report January 2011

Toxicity Test Report: TR0706/1 (page 1 of 2) This document is issued in accordance with NATA s accreditation requirements Client: VGT Pty Ltd ESA Job #: PR0706 PO Box 2335 Date Sampled: 24 January

3 Toxicity Test Report: TR0706/1 (page 1 of 2) This document is issued in accordance with NATA s accreditation requirements Client: VGT Pty Ltd ESA Job #: PR0706 PO Box 2335 Date Sampled: 24 January 2011 Greenhills NSW 2323 Date Received: 27 January 2011 Attention: Lisa Thomson Sampled By: Client Client Ref: None Supplied ESA Quote #: PL0706_q01 Lab ID No.: Sample Name: Sample Description: 4543 Triple Strike Chemical received at room temperature in apparent good condition Test Performed: Test Protocol: Test Temperature: Deviations from Protocol: Comments on Solution Preparation: Source of Test Organisms: Test Initiated: 48-hr acute (survival) toxicity test using the freshwater cladoceran Ceriodaphnia cf dubia ESA SOP 101 (ESA 2009), based on USEPA (2002) and Bailey et al. (2000) The test was performed at 25±1 C. Nil The highest test concentration was prepared by adding sample 4543 Triple Strike into dilute mineral water (DMW). The remaining test concentrations were achieved by serially diluting the highest test concentration with DMW. A DMW control was tested concurrently with the sample. ESA Laboratory culture 28 January 2011 at 1500h Sample 4543: Triple Strike Vacant Vacant Concentration (mg/l) % Survival at 48 h (Mean ± SD) DMW Control 100 ± ± ± ± ± 28.3 * ± ± 16.3 * 48-hr EC10 = 30.7mg/L** 48-hr EC50 = >1000mg/L NOEC = 37.0mg/L LOEC = 111.1mg/L *Significantly lower percent survival compared with the DMW Control (Steel s Many-One Rank Test, 1-tailed, P=0.05) **95% confidence limits are not available QA/QC Parameter Criterion This Test Criterion met? Control mean % survival >90.0% 100% Yes Reference Toxicant within cusum chart limits mg KCl/L 279.9mg KCl/L Yes

4 Toxicity Test Report: TR0706/1 (page 2 of 2) Test Report Authorised by: Dr Rick Krassoi, Director on 10 March 2011 Results are based on the samples in the condition as received by ESA. NATA Accredited Laboratory Number: This document is issued in accordance with NATA s accreditation requirements. Accredited for compliance with ISO/IEC NATA is a signatory to the APLAC mutual recognition arrangement for the mutual recognition of the equivalence of testing, calibration and inspection reports. This document shall not be reproduced except in full. Citations: Bailey, H.C., Krassoi, R., Elphick, J.R., Mulhall, A., Hunt, P., Tedmanson, L. and Lovell, A. (2000) Application of Ceriodaphnia cf. dubia for whole effluent toxicity tests in the Hawkesbury-Nepean watershed, New South Wales, Australia: method development and validation. Environmental Toxicology and Chemistry 19: ESA (2008) SOP 101 Acute toxicity test using Ceriodaphnia dubia. Issue No. 8. Ecotox Services Australasia, Sydney, New South Wales. USEPA (2002) Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms.4 th Ed. United States Environmental Protection Agency, Office of Water, Washington DC.

5 Toxicity Test Report: TR0706/2 (page 1 of 2) This document is issued in accordance with NATA s accreditation requirements Client: VGT Pty Ltd ESA Job #: PR0706 PO Box 2335 Date Sampled: 24 January 2011 Greenhills NSW 2323 Date Received: 27 January 2011 Attention: Lisa Thomson Sampled By: Client Client Ref: None Supplied ESA Quote #: PL0706_q01 Lab ID No.: Sample Name: Sample Description: 4543 Triple Strike Chemical received at room temperature in apparent good condition Test Performed: Test Protocol: Test Temperature: Deviations from Protocol: Comments on Solution Preparation: Source of Test Organisms: Test Initiated: 7-day partial life-cycle (chronic) toxicity test using the freshwater cladoceran Ceriodaphnia cf dubia ESA SOP 102 (ESA 2011), based on USEPA (2002) and Bailey et al. (2000) The test was performed at 25±1 C. Nil The highest test concentration was prepared by adding sample 4543 Triple Strike into dilute mineral water (DMW). The remaining test concentrations were achieved by serially diluting the highest test concentration with DMW. A DMW control was tested concurrently with the sample. ESA Laboratory culture 15 February 2011 at 1530h Sample 4543: Triple Strike Concentration (mg/l) Sample 4543: Triple Strike Concentration (mg/l) % Survival at 7 days (Mean ± SD) Number of Young (Mean ± SD) DMW Control 90.0 ± 31.6 DMW Control 20.8 ± ± ± ± ± ± ± ± ± 4.3 * ± ± ± ± day IC10 (survival) = ( )mg/L 7 day IC10 (reproduction) = 11.5 ( )mg/L 7 day EC50 (survival) = ( )mg/L 7 day IC50 (reproduction) = (91.4- NOEC = 111.1mg/L 156.4)mg/L LOEC = 333.3mg/L NOEC = 37mg/L LOEC = 111.1mg/L *Significantly lower percent survival compared with the DMW Control (Wilcoxon Rank Sum Test, 1-tailed, P=0.05) QA/QC Parameter Criterion This Test Criterion met? Control mean % survival >80.0% 90.0% Yes Control mean number of young > Yes Reference Toxicant within cusum chart limits mg KCl/L 235.8mg KCl/L Yes

6 Toxicity Test Report: TR0706/2 (page 2 of 2) Test Report Authorised by: Dr Rick Krassoi, Director on 10 March 2011 Results are based on the samples in the condition as received by ESA. NATA Accredited Laboratory Number: This document is issued in accordance with NATA s accreditation requirements. Accredited for compliance with ISO/IEC NATA is a signatory to the APLAC mutual recognition arrangement for the mutual recognition of the equivalence of testing, calibration and inspection reports. This document shall not be reproduced except in full. Citations: Bailey, H.C., Krassoi, R., Elphick, J.R., Mulhall, A., Hunt, P., Tedmanson, L. and Lovell, A. (2000) Application of Ceriodaphnia cf. dubia for whole effluent toxicity tests in the Hawkesbury-Nepean watershed, New South Wales, Australia: method development and validation. Environmental Toxicology and Chemistry 19: ESA (2011) ESA SOP 102 Acute Toxicity Test Using Ceriodaphnia dubia. Issue No 8. Ecotox Services Australasia, Sydney, NSW. USEPA (2002) Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms.4 th Ed. United States Environmental Protection Agency, Office of Water, Washington DC.

7 Toxicity Test Report: TR0706/3 (page 1 of 2) This document is issued in accordance with NATA s accreditation requirements Client: VGT Pty Ltd ESA Job #: PR0706 PO Box 2335 Date Sampled: 24 January 2011 Greenhills NSW 2323 Date Received: 27 January 2011 Attention: Lisa Thomson Sampled By: Client Client Ref: None Supplied ESA Quote #: PL0706_q01 Lab ID No.: Sample Name: Sample Description: 4543 Triple Strike Chemical received at room temperature in apparent good condition Test Performed: 72-hr microalgal growth inhibition test using the green alga Selenastrum capricornutum Test Protocol: ESA SOP 103 (ESA 2010), based on USEPA (2002) Test Temperature: The test was performed at 25±1 C. Deviations from Protocol: Comments on Solution Preparation: Source of Test Organisms: Test Initiated: Nil Each of the test concentrations were prepared by adding sample 4543 Triple Strike into USEPA media. The highest two concentrations were then centrifuged. A USEPA control and colour control were tested concurrently with the sample. ESA Laboratory culture, originally sourced from CSIRO Microalgal Supply Service, TAS 8 February 2011 at 1200h Sample 4543: Triple Strike Vacant Vacant Concentration (mg/l) Cell Yield x10 4 cells/ml (Mean ± SD) USEPA Control 63.0 ± 7.3 Colour Control 38.3 ± 5.0 * ± ± ± ± 0.5 ** ± 0.4 ** ± hr IC10 = 31.1 ( )mg/L 72-hr IC50 = 70.1 ( )mg/L NOEC = 37mg/L LOEC = 111.1mg/L *Significantly lower cell yield compared with the USEPA Control (Homoscedastic t Test, 1-tailed, P=0.05) **Significantly lower cell yield compared with the USEPA Control (Steel s Many-One Rank Test, 1-tailed, P=0.05)

8 Toxicity Test Report: TR0706/3 (page 2 of 2) QA/QC Parameter Criterion This Test Criterion met? Control mean cell density >16.0x10 4 cells/ml 63.9x10 4 cells/ml Yes Control coefficient of variation <20% 11.6% Yes Reference Toxicant within cusum chart limits g KCl/L 3.2g KCl/L Yes Test Report Authorised by: Dr Rick Krassoi, Director on 10 March 2011 Results are based on the samples in the condition as received by ESA. NATA Accredited Laboratory Number: This document is issued in accordance with NATA s accreditation requirements. Accredited for compliance with ISO/IEC NATA is a signatory to the APLAC mutual recognition arrangement for the mutual recognition of the equivalence of testing, calibration and inspection reports. This document shall not be reproduced except in full. Citations: ESA (2010) ESA SOP 103 Green Alga, Selenastrum capricornutum, Growth Test. Issue No 8. Ecotox Services Australasia, Sydney, NSW. USEPA (2002) Short-term methods for estimating the chronic toxicity of effluents and receiving waters to freshwater organisms. Fourth Edition. EPA-821-R United States Environmental Protection Agency, Office of Research and Development, Washington DC, USA,

9 Toxicity Test Report: TR0706/4 (page 1 of 2) This document is issued in accordance with NATA s accreditation requirements Client: VGT Pty Ltd ESA Job #: PR0706 PO Box 2335 Date Sampled: 24 January 2011 Greenhills NSW 2323 Date Received: 27 January 2011 Attention: Lisa Thomson Sampled By: Client Client Ref: None Supplied ESA Quote #: PL0706_q01 Lab ID No.: Sample Name: Sample Description: 4543 Triple Strike Chemical received at room temperature in apparent good condition Test Performed: 96-hr fish imbalance toxicity test using the rainbowfish Melanotaenia splendida Test Protocol: ESA SOP 117 (ESA 2009), based on USEPA (2002) Test Temperature: The test was performed at 25±1 C. Deviations from Protocol: Comments on Solution Preparation: Source of Test Organisms: Test Initiated: Nil The highest test concentration was prepared by adding sample 4543 Triple Strike into dilute mineral water (DMW). The remaining test concentrations were achieved by serially diluting the highest test concentration with DMW. A DMW control was tested concurrently with the sample. In-house cultures 3 March 2011 at 1400h Sample 4543: Triple Strike Vacant Vacant Concentration (mg/l) % Un-affected (Mean ± SD) DMW Control 90.0 ± ± ± ± ± ± 16.3 * ± 19.2 * 96-hr EC10 = 39.2 ( )mg/L 96-hr EC50 = ( )mg/L NOEC = 111.1mg/L LOEC = 333.3mg/L *Significantly lower percentage of un-affected larval fish compared with the DMW Control (Dunnett s Test, 1-tailed, P=0.05) QA/QC Parameter Criterion This Test Criterion met? Control mean % un-affected >80.0% 90.0% Yes Reference Toxicant within cusum chart limits µg Cu/L 12.5µg Cu/L Yes

10 Toxicity Test Report: TR0706/4 (page 2 of 2) Test Report Authorised by: Dr Rick Krassoi, Director on 10 March 2011 Results are based on the samples in the condition as received by ESA. NATA Accredited Laboratory Number: This document is issued in accordance with NATA s accreditation requirements. Accredited for compliance with ISO/IEC NATA is a signatory to the APLAC mutual recognition arrangement for the mutual recognition of the equivalence of testing, calibration and inspection reports. This document shall not be reproduced except in full. Citations: ESA (2009) SOP 117 Freshwater and Marine Fish Imbalance Test. Issue No 6. Ecotox Services Australasia, Sydney, NSW USEPA (2002) Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine organisms. Fifth edition EPA-821-R United States Environmental Protection Agency, Office of Research and Development, Washington FC, USA

11 Chain-of-Custody Documentation

12 Datasheet ID: Last Revised: 30 April 2009 Sample Receipt Notification Attention Client : Lisa Thomson : VGT PO Box 2335 Greenhills NSW lisa@vgt.com.au Telephone : (02) Facsimile : Date : 27/01/2011 Re : Receipt of Chemical Pages : 2 ESA Project : PR0706 For Review Additional Documentation Required - Please Respond FALSE Sample Delivery Details Completed Chain of Custody accompanied samples: Samples received in apparent good condition and correctly bottled: Security seals on sample bottles and esky intact: YES YES YES Date samples received : 27/01/2011 Time samples received : 11:00 No. of samples received : 1 Sample matrix : chemical Sample temperature : Comments : Includes 1L of Triple Strike (ESA ID# 4543) received at room temperature. Contact Details Customer Services Officer : Tina Micevska Telephone : Facsimile : tmicevska@ecotox.com.au Please contact customer services officer for all queries or issues regarding samples Note that the chain-of-custody provides definitive information on the tests to be performed Ecotox Services Australia ABN Phone : Unit 27, 2 Chaplin Drive Fax : Lane Cove NSW 2066 Australia info@ecotox.com.au

14 Statistical Printouts for the Acute Test with Ceriodaphnia dubia

15 Ceriodaphnia Acute Toxicity Test-48 Hr Immobilisation Start Date: 28/01/ :00 Test ID: PR0706 Sample ID: TRIPLE STRIKE End Date: 30/01/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 101 Test Species: CD-Ceriodaphnia dubia Conc-mg/L DMW Control Transform: Arcsin Square Root Rank 1-Tailed Number Total Conc-mg/L Mean N-Mean Mean Min Max CV% N Sum Critical Resp Number DMW Control * * Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates non-normal distribution (p <= 0.05) Equality of variance cannot be confirmed Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU Steel's Many-One Rank Test Treatments vs DMW Control Maximum Likelihood-Probit Parameter Value SE 95% Fiducial Limits Control Chi-Sq Critical P-value Mu Sigma Iter Slope E Intercept TSCR Point Probits mg/l 95% Fiducial Limits EC EC EC EC EC EC EC EC EC EC EC EC EC EC EC Dose mg/l Significant heterogeneity detected (p = 4.00E-02) Response Page 1 ToxCalc v Reviewed by:

16 Ceriodaphnia Acute Toxicity Test-48 Hr Immobilisation Start Date: 28/01/ :00 Test ID: PR0706 Sample ID: TRIPLE STRIKE End Date: 30/01/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 101 Test Species: CD-Ceriodaphnia dubia Dose-Response Plot Hr Immobilisation DMW Control * *1000 Page 2 ToxCalc v Reviewed by:

17 Ceriodaphnia Acute Toxicity Test-48 Hr Immobilisation Start Date: 28/01/ :00 Test ID: PR0706 Sample ID: TRIPLE STRIKE End Date: 30/01/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 101 Test Species: CD-Ceriodaphnia dubia Auxiliary Data Summary Conc-mg/L Parameter Mean Min Max SD CV% N DMW Control % Survival DMW Control ph DMW Control DO % DMW Control Cond us/cm Page 3 ToxCalc v Reviewed by:

18 Statistical Printouts for the 7-d Chronic Test with Ceriodaphnia dubia

19 Ceriodaphnia Partial Life-Cycle Test-7 Day Survival Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: Triple Strike End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Conc-mg/L DMW Control Not Fisher's 1-Tailed Number Total Conc-mg/L Mean N-Mean Resp Resp Total N Exact P Critical Resp Number DMW Control Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU Fisher's Exact Test Treatments vs DMW Control Trimmed Spearman-Karber Trim Level EC50 95% CL 0.0% % % % Auto-0.0% Response Dose mg/l Page 1 ToxCalc v Reviewed by:

20 Ceriodaphnia Partial Life-Cycle Test-7 Day Survival Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: Triple Strike End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Dose-Response Plot 7 Day Survival DMW Control Page 2 ToxCalc v Reviewed by:

21 Ceriodaphnia Partial Life-Cycle Test-7 Day Survival Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: Triple Strike End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Auxiliary Data Summary Conc-mg/L Parameter Mean Min Max SD CV% N DMW Control No of Young DMW Control % survival DMW Control ph DMW Control DO % DMW Control Cond us/cm Page 3 ToxCalc v Reviewed by:

22 Ceriodaphnia Partial Life-Cycle Test-7 Day Survival Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: Triple Strike End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Conc-mg/L DMW Control Not Fisher's 1-Tailed Isotonic Conc-mg/L Mean N-Mean Resp Resp Total N Exact P Critical Mean N-Mean DMW Control Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU Fisher's Exact Test Treatments vs DMW Control Log-Logit Interpolation (200 Resamples) Point mg/l SD 95% CL Skew IC IC IC IC IC IC IC Response Dose mg/l Page 1 ToxCalc v Reviewed by:

23 Ceriodaphnia Partial Life-Cycle Test-7 Day Survival Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: Triple Strike End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Dose-Response Plot 7 Day Survival DMW Control Page 2 ToxCalc v Reviewed by:

24 Ceriodaphnia Partial Life-Cycle Test-7 Day Survival Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: Triple Strike End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Auxiliary Data Summary Conc-mg/L Parameter Mean Min Max SD CV% N DMW Control No of Young DMW Control % survival DMW Control ph DMW Control DO % DMW Control Cond us/cm Page 3 ToxCalc v Reviewed by:

25 Ceriodaphnia Partial Life-Cycle Test-Reproduction Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: TRIPLE STRIKE End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Conc-mg/L DMW Control Transform: Untransformed Rank 1-Tailed Isotonic Conc-mg/L Mean N-Mean Mean Min Max CV% N Sum Critical Mean N-Mean DMW Control * Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates non-normal distribution (p <= 0.05) Bartlett's Test indicates equal variances (p = 0.05) Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU Wilcoxon Rank Sum Test Treatments vs DMW Control Linear Interpolation (200 Resamples) Point mg/l SD 95% CL Skew IC IC IC IC IC IC IC Response Dose mg/l Page 1 ToxCalc v Reviewed by:

26 Ceriodaphnia Partial Life-Cycle Test-Reproduction Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: TRIPLE STRIKE End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Dose-Response Plot Reproduction DMW Control * Page 2 ToxCalc v Reviewed by:

27 Ceriodaphnia Partial Life-Cycle Test-Reproduction Start Date: 15/02/ :30 Test ID: PR0706/11 Sample ID: TRIPLE STRIKE End Date: 22/02/ :30 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 102 Test Species: CD-Ceriodaphnia dubia Auxiliary Data Summary Conc-mg/L Parameter Mean Min Max SD CV% N DMW Control No of Young DMW Control % survival DMW Control ph DMW Control DO % DMW Control Cond us/cm Page 3 ToxCalc v Reviewed by:

28 Statistical Printouts for the Selenastrum Growth Inhibition Tests

29 Microalgal Growth inhibition Test-Growth-Cell Yield Start Date: 8/02/ :00 Test ID: PR0706/11 Sample ID: Controls End Date: 11/02/ :00 Lab ID: NA Sample Type: AQ-Aqueous Sample Date: Protocol: ESA 103 Test Species: SC-Selenastrum capricornutum Conc-% USEPA Control Colour Control Transform: Untransformed 1-Tailed Conc-% Mean N-Mean Mean Min Max CV% N t-stat Critical MSD USEPA Control *Colour Control Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.05) F-Test indicates equal variances (p = 0.56) Hypothesis Test (1-tail, 0.05) MSDu MSDp MSB MSE F-Prob df Homoscedastic t Test indicates significant differences E E , 6 Treatments vs USEPA Control Dose-Response Plot Growth-Cell Yield tail, 0.05 level of significance USEPA Control *Colour Control Page 1 ToxCalc v Reviewed by:

30 Microalgal Growth inhibition Test-Growth-Cell Yield Start Date: 8/02/ :00 Test ID: PR0706/11 Sample ID: Controls End Date: 11/02/ :00 Lab ID: NA Sample Type: AQ-Aqueous Sample Date: Protocol: ESA 103 Test Species: SC-Selenastrum capricornutum Auxiliary Data Summary Conc-% Parameter Mean Min Max SD CV% N USEPA Control Cell Yield Colour Control USEPA Control ph Colour Control USEPA Control Cond us/cm Colour Control Page 2 ToxCalc v Reviewed by:

31 Microalgal Growth inhibition Test-Growth-Cell Yield Start Date: 8/02/ :00 Test ID: PR0706/10 Sample ID: Triple Strike End Date: 11/02/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 103 Test Species: SC-Selenastrum capricornutum Conc-mg/L USEPA Control Transform: Untransformed Rank 1-Tailed Isotonic Conc-mg/L Mean N-Mean Mean Min Max CV% N Sum Critical Mean N-Mean USEPA Control * * Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.05) Bartlett's Test indicates unequal variances (p = 4.20E-04) Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU Steel's Many-One Rank Test Treatments vs USEPA Control Linear Interpolation (200 Resamples) Point mg/l SD 95% CL(Exp) Skew IC IC IC IC IC IC IC Response Dose mg/l Page 1 ToxCalc v Reviewed by:

32 Microalgal Growth inhibition Test-Growth-Cell Yield Start Date: 8/02/ :00 Test ID: PR0706/10 Sample ID: Triple Strike End Date: 11/02/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 103 Test Species: SC-Selenastrum capricornutum Dose-Response Plot Growth-Cell Yield USEPA Control *111.1 * Page 2 ToxCalc v Reviewed by:

33 Microalgal Growth inhibition Test-Growth-Cell Yield Start Date: 8/02/ :00 Test ID: PR0706/10 Sample ID: Triple Strike End Date: 11/02/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 103 Test Species: SC-Selenastrum capricornutum Auxiliary Data Summary Conc-mg/L Parameter Mean Min Max SD CV% N USEPA Control Cell Yield USEPA Control ph USEPA Control Cond us/cm Page 3 ToxCalc v Reviewed by:

34 Statistical Printouts for the Larval Fish Imbalance Tests

35 Fish Imbalance Test-96 hr Imbalance Start Date: 3/03/ :00 Test ID: PR0706/10 Sample ID: Triple Strike End Date: 7/03/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 117 Test Species: MS-Melanotaenia splendida Conc-mg/L DMW Control Transform: Arcsin Square Root 1-Tailed Number Total Conc-mg/L Mean N-Mean Mean Min Max CV% N t-stat Critical MSD Resp Number DMW Control * * Auxiliary Tests Statistic Critical Skew Kurt Shapiro-Wilk's Test indicates normal distribution (p > 0.05) Bartlett's Test indicates equal variances (p = 0.89) Hypothesis Test (1-tail, 0.05) NOEC LOEC ChV TU MSDu MSDp MSB MSE F-Prob df Dunnett's Test E-05 6, 21 Treatments vs DMW Control Maximum Likelihood-Probit Parameter Value SE 95% Fiducial Limits Control Chi-Sq Critical P-value Mu Sigma Iter Slope Intercept TSCR Point Probits mg/l 95% Fiducial Limits 0.9 EC EC EC EC EC EC EC EC EC EC EC EC EC E+09 EC EC E+08 Dose mg/l Response Page 1 ToxCalc v Reviewed by:

36 Fish Imbalance Test-96 hr Imbalance Start Date: 3/03/ :00 Test ID: PR0706/10 Sample ID: Triple Strike End Date: 7/03/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 117 Test Species: MS-Melanotaenia splendida Dose-Response Plot hr Imbalance tail, 0.05 level of significance DMW Control *333.3 *1000 Page 2 ToxCalc v Reviewed by:

37 Fish Imbalance Test-96 hr Imbalance Start Date: 3/03/ :00 Test ID: PR0706/10 Sample ID: Triple Strike End Date: 7/03/ :00 Lab ID: 4543 Sample Type: CP-Chemical product Sample Date: Protocol: ESA 117 Test Species: MS-Melanotaenia splendida Auxiliary Data Summary Conc-mg/L Parameter Mean Min Max SD CV% N DMW Control % Unaffected DMW Control ph DMW Control Cond us/cm DMW Control DO % Page 3 ToxCalc v Reviewed by:

Toxicity Assessment of Hydragyp. VGT Pty Ltd. Test Report

Toxicity Assessment of Hydragyp VGT Pty Ltd Test Report September 2010 Toxicity Assessment of Hydragyp VGT Pty Ltd Test Report September 2010 Toxicity Test Report: TR0627/1 (page 1 of 2) This document