ISO Common Name Thiamethoxam. Company Code: CGA 293343 Tech

RMS: Spain Thiamethoxam Doc III-A

98/8 Doc IIIA 7.4.1.1 / Acute toxicity to fish section No. 01 91/414 Annex II Acute toxicity to fish Point addressed 8.2.1 / 03

1. Annex point(s) II A, 8.2.1 Acute Toxicity to Fish 2. Location in Dossier Section 6 3. Authors / Year

Title 1 ACUTE TOXICITY TEST OF CGA 293343 TECH. TO RAINBOW TROUT (ONCORHYNCHUS MYKISS) IN THE FLOW-THROUGH Report No. / Date SYSTEM Novartis File N° 95R002 / 30.01.1996

Source / Owner Novartis Study # 293343-36

Unpublished / Novartis Crop Protection AG

4. Testing facility

5. Dates of work September 15, 1995 until December 6, 1995 6. Test substance ISO common name thiamethoxam. Company Code: CGA 293343 tech.,

7. Test method OECD Guideline No.: 203; U.S. EPA FIFRA Pesticide Assessment Guidelines, Subdivision E, Section No. 72-1; 92/69/EEC C.1. 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: Thiamethoxam technical; Juvenile Rainbow trout, Oncorhynchus mykiss, (mean body length 53 mm; mean body weight 135 g) were exposed to one concentration of the test substance (nominally 100 mg/L) under flow- through conditions for 96 hours. Seven fish randomly selected were employed in the treatment group and the water control group with one test chamber each. The compound appeared to be homogeneously distributed in the test vessels at all observation times and concentrations, however, at the start of the test the formation of small clumps clogging the valve of the pump lead to the high variation of the test substance concentration measured during the first 24 hours of the study. Measured concentrations of thiamethoxam determined daily ranged from 39 to 243% of nominal during the first 24 hours and from 108 to 125 % during the following 72 hours of the test. Taking into account the irregular dosing at the start of the test, the compound seemed to be stable in the test solution over the period of the study under flow-through conditions. Results are based on the mean measured concentration of 125 mg a.i./L.

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Findings: Acute toxicity of thiamethoxam to Rainbow trout

Concentration Mortality Exposure LCs o• (mg a.i./L] after 96 h period (mg a.i./L] (%] (hours] (95 % cont. interval) Nominal Mean measured control 0 0 24h > 125 100 125 0 96 h > 125 NOEC: 125 mg a.i./L

•calculated based on measm·ed concentrations

Observations: No sublethal effects, such as a change in pigmentation, loss of equilibrium, change in the swimming behaviour or in the respiratory function were observed at any time of the study.

Conclusion: The acute 96-hour LC50 for Rainbow trout exposed under flow-through conditions was detennined to be > 125 mg a.i./L.

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Results and discussion -

Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability

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Acceptability Remarks

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98/8 Doc IIIA 7.4.1.1 / Acute toxicity to fish section No. 02 91/414 Annex II Acute toxicity to fish Point addressed 8.2.1 / 05

1. Annex point(s) II A, 8.2.1 Acute Toxicity to Fish 2. Location in Dossier Section 6 3. Authors / Year

Title 2 ACUTE TOXICITY TEST OF CGA 293343 TECH. TO RAINBOW TROUT (ONCORHYNCHUS MYKISS) UNDER FLOW-THROUGH CONDITIONS Report No. / Date 972548 / 26.11.1997 Novartis File N° Novartis Study # 293343-388 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility . 5. Dates of work June 10, 1997 through June 14, 1997 6. Test substance ISO common name thiamethoxam. Company Code: CGA 293343 tech.,

7. Test method OECD Guideline No.: 203; U.S. EPA FIFRA Pesticide Assessment Guidelines, Subdivision E, Section No. 72-1; 92/69/EEC C.1. (Total Length: mean 54 mm, (range 49- 62 mm) based on 20 fish, one fish out of 20 was exceeding the OECD range of 40-60 mm) 8. Deviations none 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: Thiamethoxam technical; Juvenile Rainbow trout, Oncorhynchus mykiss, (mean body length 54 mm; mean body weight 139 g) were exposed to one concentration of the test substance (nominally 100 mg/L) under flow- through conditions for 96 hours. Thirty fish randomly selected were employed in the treatment group (3 replicates of 10 fish) and 20 fish in the water control group (2 replicates of 10) with 10 fish per test chamber. The compound was homogeneously distributed in the test vessels at all observation times and concentrations. No precipitation was observed in the stock solution or the testing solutions throughout the duration of the test. Measured concentrations of thiamethoxam determined daily ranged from 89-120% of nominal concentrations over the period of the test. The mean measured concentration was 107 mg ai/l. Results are based on nominal values.

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Findings: Acute toxicity of thiamethoxam to Rainbow trout

Concentration Mortality Exposure LC so a (mg a.i./L] after 96 h period (mg a.i./L] (%] (hours] (95 % cont. interval) Nominal Mean measured control 0 0 24h > 100 100 107 0 96-h > 100 NOEC: 100 mg a.i./L

•calculated based on nominal concentrations

Observations: No sublethal effects, such as a change in pigmentation, loss of equilibrium, change in the swimming behaviom or in the respiratory function were observed at any time of the study.

Conclusion: The acute 96-hour LC50 for Rainbow trout exposed under flow-through conditions was detemlined to be > 100 mg a.i./L.

Results and discussion Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and M ethods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc lllA 7.4.1.1 Acute toxicity to fish section No. 03 91 /414 Annex II Acute toxicity to fish Point addressed 8.2.1 I 02

1. Annex point(s) II A, 8.2.1 Acute Toxicity to Fish 2. Location in Dossier Section 6 3. Authors I Year

Title 3 CGA-293343 A 96-HOUR FLOW-THROUGH ACUTE TOXICITY TEST WITH THE BLUEGILL, LEPOMIS MACROCHIRUS Report No. I Date 108A-189 I 14.10.1996

Novartis File N° Novartis Study# 293343-145

Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility 5. Dates of work September 5, 1996 through September 9, 1996 6. Test substance ISO common name thiamethoxam.

7. Test method U.S. EPA FIFRA Pesticide Assessment Guidelines, Subdivision E, Section No.72-1 (1982), and ASTM Standard E 729-88a. 8. Deviations None 9. GLP The study was conducted to conform with Good Laborato1y Practice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989; OECD, ISBN 92-84-12367-9, Paris 1982; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: Thiamethoxam technical; Bluegill sunfish (Lepomis macrochirus) (mean o y engt mm; mean o y we1 t . g were exposed to five concentrations of the test substance (nominal test concentrations of 16, 26, 43, 72 and 120 mg a.i./L) under flow-through conditions for a period of 96 hours. Two groups of ten fish randomly selected were employed per concentration and the water control. After incorporating the test substance into the diluter system, test solutions appeared clear and colourless. Measured test concentrations of thiamethoxam ranged from 81-96% of the nominal values during the period of exposure. The mean measured concentrations were 14, 24, 40, 64 and 114 mg test substance/I. The following values are based on mean measured concentrations.

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Findings: Acute toxicity of thiamethoxam to Bluegill sunfish Concentration Mortality Exposure LCso " (mg a.i./L] after 96 h period (mg a.i./L] (%] (hours] (95 % cont. interval) Nominal Mean measured control 0 0 24h > 114 16 14 0 48 h > 114 26 24 0 72h > 114 43 40 0 96 h > 114 72 64 0 120 114 0 NOEC: 114 mg a.i./L a calculated based on measured concentrations

Observations: No mo1talities were obse1ved and fish appeared n01mal throughout the test with no ove1t signs of toxicity or any other sublethal effects.

Conclusion: The acute 96-hour LC50 for Bluegill sunfish exposed to thiamethoxam under flow-through conditions was dete1mined to be > 114 mg a.i./L.

Evaluation by Competent Authorities 7.4.1.1./03

108A-189

EVALUATION BY RAPPORTEUR MEMBER STATE Date 09/02/2005 Materials and Methods

Results and discussion

Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Give date ofcomments submitted

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Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summary and conclusion. Discuss if deviating from view of rapporteur member state Results and discussion Discuss if deviating from view of rapporteur member state Conclusion Discuss if deviating from view of rapporteur member state Reliability Discuss if deviating from view of rapporteur member state Acceptability Discuss if deviating from view of rapporteur member state Remarks

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98/8 Doc IIIA 7.4.1.1 / Acute toxicity to fish section No. 04 91/414 Annex II Acute toxicity to fish Point addressed 8.2.1 / 06

1. Annex point(s) II A, 8.2.1 Acute Toxicity to Fish 2. Location in Dossier Section 6 3. Authors / Year

Title 4 ACUTE TOXICITY TEST OF CGA 322704 (METABOLITE OF CGA 293343) TO RAINBOW TROUT (ONCORHYNCHUS MYKISS) IN THE STATIC SYSTEM Report No. / Date 962527 / 21.01.1997 Novartis File N° Novartis Study # 322704-9 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility

5. Dates of work August 19, 1996 through November 5, 1996 6. Test substance Company Code: CGA 322704,

7. Test method OECD Guideline No.: 203; 92/69/EEC C.1. U.S. EPA FIFRA Pesticide Assessment Guidelines, Subdivision E, Section No. 72-1; (Deviation from FIFRA -Guideline: Number of replicates of test organisms, Continuous monitoring of temperature and equilibration time) 8. Deviations none 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: CGA 322704, The test was performed as a limit test. Juvenile Rainbow trout, Oncorhynchus mykiss, (mean body length 53 mm; mean body weight 13 g) were exposed to one concentration of the test substance (nominally 100 mg/L) under static test conditions for 96 hours. Seven fish per test concentration, and control were added to each aquarium. Measured concentrations of CGA 322704 determined at the beginning and the end of the test ranged from 72% to 102% of nominal values. Based on the dissolved fraction in the samples the measured concentrations were 70% and 100% of the nominal values, respectively. However, a precipitate was observed at 100 mg ai/l from the start of the test until 24h of exposure which lead to a 70% recovery in the dissolved fraction at the beginning of exposure. After 24 hours this precipitate was completely dissolved resulting in 100% recovery at the end of the exposure period. Therefore, the following values are based on nominal concentrations.

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Findings: Acute toxicity of CGA 322704 (metabolite of thiamethoxam) to Rainbow trout

a Concentration of CGA 322704 Mortality Exposure LC50 [mg /L] after 96 h period [mg/L] [%] [hours] (95 % conf. interval) Nominal Mean measured control 0 0 100 87 0 96 h > 100 NOEC: 100 mg/L a based on nominal values

Observations: No sublethal effects, such as a change in pigmentation, loss of equilibrium, change in the swimming behaviour or in the respiratory function were observed at any time of the study.

Conclusion: The acute 96-hour LC50 for Rainbow trout exposed to CGA 322704 (metabolite of thiamethoxam) under static conditions was determined to be >100 mg /L.

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Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 09/03/05 Materials and Methods

Results and discussion

Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc IIIA 7.4.1.1 / Acute toxicity to fish section No. 05 91/414 Annex II Acute toxicity to fish Point addressed 8.2.1 / 02

1. Annex point(s) II A, 8.2.1 Acute Toxicity to Fish 2. Location in Dossier Section 6 3. Authors / Year

Title 5 ACUTE TOXICITY OF CGA 355190 (METABOLITE OF CGA 293343) FOR RAINBOW TROUT (DETERMINATION OF THE LC-VALUES) Report No. / Date G 54104, 982588 / 29.10.1998 Novartis File N° Novartis Study # 355190-2 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility 5. Dates of work September 7, 1998 through September 30, 1998 6. Test substance Company Code: CGA 355190,

7. Test method OECD Guideline No.: 203; 92/69/EEC C.1. 8. Deviations none 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: CGA 355190, The test was performed as a limit test. Juvenile Rainbow trout, Oncorhynchus mykiss, (mean body length 42 mm; mean body weight 0.68 g) were exposed to one concentration of the test substance (nominally 100 mg/L) under static test conditions for 96 hours. Seven fish per test concentration and control were added to each aquarium. Measured concentrations of CGA 355190 determined at the beginning and the end of the test ranged from 95.1% to 96.9% of nominal values. Therefore, the following values are based on nominal concentrations.

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Findings: Acute toxicity of CGA 355190 (metabolite of thiamethoxam) to Rainbow trout

a Concentration of CGA 355190 Mortality Exposure LC50 [mg /L] after 96 h period [mg/L] [%] [hours] (95 % conf. interval) Nominal Mean measured control < 0.4 0 100 95.8 0 96 h > 100 NOEC: 100 mg/L a based on nominal values

Observations: No sublethal effects, such as a change in pigmentation, loss of equilibrium, change in the swimming behaviour and in the respiratory function were observed at any time of the study.

Conclusion: The acute 96-hour LC50 for Rainbow trout exposed to CGA 355190 (metabolite of thiamethoxam) under static conditions was determined to be >100 mg /L.

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Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 10/03/05 Materials and Methods

Results and discussion

Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc IIIA 7.4.1.1 / Acute toxicity to fish section No. 06 91/414 Annex II Acute toxicity to fish Point addressed 8.2.1 / 07

1. Annex point(s) II A, 8.2.1 Acute Toxicity to Fish 2. Location in Dossier Section 6 3. Authors / Year

Title 6 ACUTE TOXICITY OF NOA 407475 (METABOLITE OF CGA 293343) TO RAINBOW TROUT (ONCORHYNCHUS MYKISS) IN A 96-HOUR STATIC Report No. / Date TEST 688781 / 14.08.1998 Novartis File N° Novartis Study # 407475-10 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility 5. Dates of work July 06, 1998 through July 22, 1998 6. Test substance Company Code: NOA 407475,

7. Test method OECD Guideline No.: 203; 92/69/EEC C.1. 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: NOA 407475, The test was performed as a Limit test. Juvenile Rainbow trout, Oncorhynchus mykiss, (mean body length 46 ± 0.2 mm; mean body weight 1.00 ± 0.2 g) were exposed to one concentration of the test substance (nominally 100 mg/L) under static test conditions for 96 hours. Seven fish per test concentration, and control were added to each aquarium. Measured concentrations of NOA 407574 determined at the beginning and the end of the test were 87 % of nominal values. Therefore, the following values are based on nominal concentrations.

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Findings: Acute toxicity of NOA 407475 (metabolite of thiamethoxam) to Rainbow trout

a Mortality Exposure LC50 Concentration of NOA 407475 after 96 h period [mg/L] [mg /L] [%] [hours] (95 % conf. interval) Nominal Mean measured control < 0.4 0 100 87.0 0 96 h > 100 NOEC: 100 mg/L a based on nominal values

Observations: No sublethal effects, such as a change in pigmentation, loss of equilibrium, change in the swimming behaviour or in the respiratory function were observed at any time of the study.

Conclusion: The acute 96-hour LC50 for Rainbow trout exposed to NOA 407475 (metabolite of thiamethoxam) under static conditions was determined to be >100 mg /L.

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Evaluation by Competent Authorities 7.4.1.1/06

688781

EVALUATION BY RAPPORTEUR MEMBER STATE Date 10/03/05 Materials and Methods

Results and discussion

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98/8 Doc lllA 7.4.1.1 Acute toxicity to fish section No. 7 91 /414 Annex II Acute toxicity to fish Point addressed 8.2.1

1. Annex point(s) II A, 8.2.1 Acute Toxicity to Fish 2. Location in Dossier Section 6 3. Authors I Year

Title 7 NOA 459602 (THIAMETHOXAM METABOLITE): ACUTE TOXICITY TEST TO RAINBOW TROUT (ONCORHYNCHUS MYKISS). Report No. I Date 01 -0416/B I 14.08.1998

Novartis File N° 459602/0016

Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility 5. Dates of work July 06, 1998 through July 22, 1998 6. Test substance

7. Test method OECD - Guideline No.: 203, Paris 1992; 92/69/EEC C.1. 8. Deviations None 9. GLP Yes (certified laborato1y)

Test System: NOA 459602 (metabolite of thiamethoxam), Rainbow trout, Oncorhynchus mykiss, (source: control fish at test end: mean body length 57 mm range 55 to 59 mm ; mean o y Ml43 g (range 1.36 to 3.71 g) were exposed to a single concentration of the test item under static conditions for a pe1iod of 96 hours. The test concentration (nominal: 120 mg NOA 459602 per litre) was evaluated. For the test concentration a single stock solution was prepared without using a solvent. In addition to the test item treatment a blank water control was tested. One group of 10 fish randomly selected were maintained in the treatment and control groups. The test was pe1fo1med in 40 L-glass aqua1ia containing 25 L of test solution. Loading rate was approximately 0.97 g fish/L (test end in control). Mo1talities and symptoms of toxicity were recorded after 3, 24, 48, 72 and 96 hours. Temperature, dissolved oxygen and pH were measured daily in each test chamber. Total hardness of the water was dete1mined pe1iodically at the laborato1y. Samples of test media from the test item and control aquaria were taken before test strut and after 96 hours for the analytical dete1mination (HPLC) of the test concentrations. Findings: Results are given on basis of nominal concentrations. Concentrations va1ied between 100% of nominal at test stait and 108% of nominal at test end. The overall mean concentration was 108% of nominal. The water temperature was in the range of 14.9 to 15.3 °C and a lighting regime of 16 hours light and 8 hours dark per day was employed. The dissolved oxygen content vai·ied between 97 and 102 % of saturation, pH was 7.39 - 7.76, at test stait the total hai·dness was 45.0 mg CaC03/L and the conductivity was 209 µSiem.

Table 1 Acute toxicity of NOA 459602 (metabolite of thiamethoxam) to rainbow trout I Mortality I Exposure I LCso 5

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Concentration after 96 h period (95 % conf. (mg NOA 459602 I L] (%] (hours] interval) (mg NOA 459602 I LI Nominal Sta1t End - Control <0.75 <0.75 0 120 120 130 0 96h > 120 (n.a.) NOEC: 120 mg NOA 459602 I L

a based on nominal concentrations; n.a. = not applicable

Observations: No mortalities or sublethal effects were observed at any time of the study.

Conclusion: The acute 96-hour LC50 for Rainbow trout exposed to NOA 459602 (metabolite of thiamethoxam) under static conditions was detennined to be > 120 mg /L.

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 10/03/05 Materials and Methods

Results and discussion Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviatin g from view ofra pporteur member state Results and discussion Discuss ifdeviatin g from view ofra pporteur member state Conclusion Discuss ifdeviating from view ofra pporteur member state Reliability Discuss ifdeviating from view ofra pporteur member state Acceptability Discuss ifdeviating from view ofra pporteur member state Remarks

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98/8 Doc lllA 7.4.1.2 Acute toxicity to invertebrates section No. 01 91 /414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4 / 04

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Inve1·tebrates 2. Location in Dossier Section 6 3. Authors I Year Neumann, C. (1996)

Title 8 ACUTE TOXICITY TEST OF CGA 293343 TO THE CLADOCERAN DAPHNIA MAGNA STRAUS UNDER STATIC CONDITIONS Report No. I Date 950003 1 25.04.1996

Novartis File N° Novartis Study# 293343-43

Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection, Inc. Ecotoxicology Laboratories, CH-4002 Basie Switzerland 5. Dates of work September 5, 1995 through September 7, 1995 6. Test substance ISO common name thiamethoxam.

7. Test method OECD Guideline No.: 202, Paris 1984 and FIFRA Guideline No.: 72-2, December 24, 1989. 8. Deviations None 9. GLP The study was conducted to conform with Good Laborato1y Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989 1) OECD Principles of Good Laborato1y, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: Thiamethoxam technical, The acute toxicity of thiamethoxam to Daphnia magna has been etemune wit ive concentrat10ns of test substance and a blank control at a temperature of 20 ±2 °C. Daphnids were exposed to the test concentrations under static conditions. For each concentration and the control, four replicates were set up. Each replicate consisted of 5 daphnids in 100 mL test medium (M4 medium). The daphnids were observed for immobilisation and behavioural changes after 24 h and 48 h of exposure. Nominal concentrations of thiamethoxam were 10, 18, 32, 58 and 100 mg/L. Concentrations measured at the test initiation were 9.92, 19.46, 35.85, 62.23, and 109.5 mg/L. At test termination measured concentrations were 11 .15, 18.34, 37.05, 58.78 and 102.1 mg/L. Since the test substance was stable under the experimental conditions and constant exposure maintained throughout the static exposure, results are based on nominal concentrations.

Findings: Acute toxicity of thiamethoxam to Dapnia magna Concentration Immobilisation Exposure ECsoa (mg a.i./L] after 48 h period (mg a.i./L] (%] (hours] (95 % cont. interval)

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Nominal Measured End controls 0 0 24h > 100 10 11.15 0 18 18.34 0 48 h > 100 32 37.05 0 58 58.78 5 100 102.1 15 NOEC: 32 mg a.i./L

• calculated based on nominal concentrations

Observations: A low percentage of immobilisation was recorded only at the highest concentrations of 58 and 100 mg a.i./L No fmther effects on daphnids were obse1ved.

Conclusion: The acute 48-hour EC50 of Daphnia magna exposed to thiamethoxam under static conditions was dete1m ined to be > 100 mg a.i./L.

Results and discussion

Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 02 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors (Year) Knauer, K. (2000b)

Title 9 Acute toxicity test of CGA 293343 tech. to Gammarus Sp. under static Report No. / Date conditions 2002614 / 10.07.2000 Syngenta File N° 293343-1229 Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Syngenta Crop Protection AG, Basel Switzerland 5. Dates of work 9th June 2000 through 17th June, 2000 6. Test substance ISO common name thiamethoxam; Company Code: CGA 293343 tech.;

7. Test method OECD-Guideline No.: 202, Paris 1984 EEC-C.2, December 29, 1992 U.S. EPA-FIFRA Guideline Number 72-2, December 24, 1989 U.S. EPA OPPTS Test Guideline OPPTS 850.1010, April, 1996 8. Deviations None 9. GLP GLP with the exception of range finding test the study was conducted in compliance with the Siss Ordinance relating to Good Laboratory Practice, adopted 2nd February, 2000 (RS 813.016.5). This ordinance is based upon the OECD Principles of Good Laboratory Practice, as revised in 1997 and adopted 26th November, 1997 by decision of the OECD Council [C (97) 186/Final]. These procedures are based on the OECD Principles of Good Laboratory Practice adopted 12th May, 1981 by the decision of the OECD Council [C (81) 30 (Final)] concerning Mutual Acceptance of Data in the Assessment of Chemicals, as ammended by the Council Decision-Recommendation of 2 October, 1989 concerning Compliance with Principles of Good Laboratory Practice Council [C (89) 87 (Final)].

Test System: Thiamethoxam technical, . The acute toxicity of thiamethoxam (technical) to the freshwater crustacean Gammarus Sp. has been determined at six different test concentrations and a blank control at a temperature of 20 ± 2 °C. The test organism, Gammarus Sp. was collected from the River Kastelbach, Kaltbrunnental, Switzerland, then transferred, together with the natural river water, to an aquarium housed in the Novartis laboratories. The single species required for testing was accordingly separated. Gammarus Sp. was acclimatised for at least a period of 20-h. No feed was provided during the 48-h test period. A range-finding test was performed at four concentrations (0.1, 1, 10 and 100 mg/L nominal) prior to embarking upon the definitive study phase. Based on results from the range-finder, the definitive test was conducted at 0 (Control), 1.6, 3.1, 6.3, 13, 25 and 50 mg thiamethoxam/L. Analysis of fortified test solutions at 0-hr and 48-hrs indicated that measured exposure concentrations were, respectively, in the range 93-99% and 92-96% of the nominal concentrations. Four replicates per test concentration containing 5 organisms per replicate were prepared in 250 mL glass test vessels containing 200 mL of test solution (CGA 293343 tech. dissolved in river water). Test vessels containing Gammarus Sp. were incubated in a climatic chamber for a period of 48-hours and subjected to a 16 hour light / 8 hour dark light regime, employing a 30 minute transition period. Average light intensity, during periods of illumination, was the order of 24 µE/m2sec. Water quality measurements included pH (range 8.3-8.5), temperature (constant at 21 °C) and oxygen content (range 65-101 % of

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Findings: No immobilisation was observed in the control (blank pond water medium) during the course of the 48-hour exposure period, with no unexpected phenomena being observed. The test was thus deemed valid. The results are presented below. Acute toxicity ofthiamethoxam (tech.) to Gammarus

Concentration Immobilisation Exposure ECso0 (mg a.i./L] after 48 h period (mg a.i./L] (%] (hours] (95 % cont. interval) Nominal Measured Beginning / End control < 0.01 I< 0.01 0 24 h 15 1.6 1.57 I l.56 40 3. 1 3.07 / 2.94 50 6.3 6.13 / 5.85 60 48 h 2.8 13 12.1 / 12.5 95 25 24.0 / 23.1 95 50 47.4 I 46.7 100 NOEC" : < 1.6 mg a.i./L

• calculated based on nominal concentrations " Highest concentration with no immobilisation or behavioural changes

Conclusion: The acute 48-hour EC50 for the Gammarus Sp. when exposed to thiamethoxam under static conditions was determined to be 2.8 mg a.i./L. The 48-hour NOEC was determ ined to be less than 1.6 m a.i./L, the lowest concentration level tested. Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the cormnents and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 24/11/2004 Materials and Methods

Results and discussion

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Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Give date ofcomments submitted Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 03 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors (Year) Knauer, K. (2000c)

Title 10 Acute toxicity test (24h) of CGA 293343 tech. to three invertebrate species Daphnia pulex Leydig, Thamnocephalus platyurus, and Brachionus calyciflorus under Report No. / Date static conditions. 2002612 / 21.07.2000 Syngenta File N° 293343/1274 Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Syngenta Crop Protection AG, Basel Switzerland 5. Dates of work 6th June 2000 through 15th June, 2000 6. Test substance ISO common name thiamethoxam; Company Code: CGA 293343 tech.,

Test System: Thiamethoxam technical, The acute toxicity of thiamethoxam (technical) to three individual invertebrate species (Daphnia pulex Leydig, Thamnocephalus platyurus both of which are freshwater crustaceans, and Brachionus calyciflorus, a freshwater rotifer species) has been determined under static conditions at six different test concentrations, plus a blank control, at a temperature of between 21 and 24 °C. A 24-hour exposure period was employed for the definitive test in view of the rapid metabolic characteristics of the test species. Additional prolongation could potentially result in starvation. Daphnia pulex and Thamnocephalus platyurus (both genus Daphnia) and Brachionus calyciflorus (genus Brachionus) were obtained from a reliable commercial supplier. The test animals were derived from cysts and ephippia hatched in the laboratory according to the given standard operating procedure. The acclimation period lasted for at least 2 hours, not exceeding 48 hours, and for Brachionus not in excess of 32 hours. The hatched organisms were maintained in a “moderately hard” reconstituted water (in-line with the formula recommended by the US EPA) and adjusted to pH 8.5. Stock solutions of thiamethoxam were prepared in “moderately hard” reconstituted water medium. A range-finding test was performed at three concentrations (1, 10 and 100 mg/L nominal) prior to embarking upon the definitive study phase. Based on results from the range- finder, the definitive test was conducted at 0 (Control), 3.1, 6.3, 12.5, 25, 50 and 100 mg thiamethoxam/L. Analysis of fortified test solutions at 0-hr and 48-hrs indicated that measured exposure concentrations

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were, respectively, in the range 97-116% and 96-113% of the nominal concentrations for the test conducted with Daphnia pulex, and, 95-118% and 95-117% of nominal over the same exposure period for both Thamnocephalus and Brachionus. A total of 20, 30 and 30 organisms, respectively for Daphnia pulex Leydig, Brachionus calcyciflorus and Thamnocephalus platyurus, for each test concentration and the control, constituting, respectively, four, three and six replicates at each test concentration and control were performed. Multiwell plates were adopted as test vessels for the test. The following volumes were correspondingly used: Daphnia pulex, 5 mL; Thamnocephalus platyurus, 1 mL; Brachionus calyciflorus, 0.3 mL. Test vessels were incubated in a climatic chamber regulated at 21 to 24 °C. The test was performed in the dark. Water quality measurements included the following parameters: pH (range 8.0- 8.3), temperature (21-22 °C for Daphnia pulex test, 23-24 °C for the two remaining species), oxygen content (93-101 % of saturation), hardness (82-84 mg CaCO3/L), and conductivity (301-318 µS/cm). Immobilisation or other behavioural changes were recorded after 24-hours.

Findings: No immobilisation was observed in the control (blank culture medium) during the course of the 24-hour exposure period, with no unexpected phenomena being observed. The test was thus deemed valid. The results of the acute tests performed with Daphnia pulex, Thamnocephalus platyurus and Brachionus calcyiflorus are presented below.

Acute toxicity of thiamethoxam (tech.) to Daphnia pulex

a Concentration Immobilisation Exposure EC50 [mg a.i./L] after 24 h period [mg a.i./L] [%] [hours] (95 % conf. interval) Nominal Measured Beginning / End control < 0.01 / < 0.01 0 3.1 3.12 / 3.08 0 6.3 7.28 / 7.05 10 12.5 12.6 / 12.8 15 24 h > 100 25 24.9 / 24.2 25 50 48.7 / 47.9 10 100 97.6 / 98.3 20 NOEC^: 6.3 mg a.i./L a calculated based on nominal concentrations ^ Highest concentration with ≤ 10% immobilisation or behavioural changes (NB - up to 10% immobilisation is considered acceptable in the control to fulfill test validity criteria)

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Acute toxicity of thiamethoxam (tech.) to Thamnocephalus platyurus

a Concentration Immobilisation Exposure EC50 [mg a.i./L] after 24 h period [mg a.i./L] [%] [hours] (95 % conf. interval) Nominal Measured Beginning / End control < 0.01 / < 0.01 0 3.1 3.13 / 3.13 3.3 6.3 7.38 / 7.34 3.3 12.5 12.5 / 12.2 3.3 24 h > 100 25 23.8 / 24.5 0 50 48.3 / 47.4 3.3 100 101.3 / 98.5 3.3 NOEC^: 100 mg a.i./L a calculated based on nominal concentrations ^ Highest concentration with ≤ 10% immobilisation or behavioural changes (NB - up to 10% immobilisation is considered acceptable in the control to fulfill test validity criteria) Acute toxicity of thiamethoxam (tech.) to Brachionus calyciflorus

a Concentration Immobilisation Exposure EC50 [mg a.i./L] after 24 h period [mg a.i./L] [%] [hours] (95 % conf. interval) Nominal Measured Beginning / End control < 0.01 / < 0.01 0 3.1 3.13 / 3.13 0 6.3 7.38 / 7.34 0 12.5 12.5 / 12.2 0 24 h > 100 25 23.8 / 24.5 0 50 48.3 / 47.4 6.7 100 101.3 / 98.5 0 NOEC^: 100 mg a.i./L a calculated based on nominal concentrations ^ Highest concentration with ≤ 10% immobilisation or behavioural changes (NB - up to 10% immobilisation is considered acceptable in the control to fulfill test validity criteria)

Observations: The EC50 was not attained for any of the three species tested, and thus is cited as > 100 mg a.i./L, where 100 mg a.i./L represents the highest test concentration studied. With reference to the accepted validity criteria set for Control samples, which permits up to a maximum of 10 % immobilisation over the period of the test and applying this principle to the various test concentrations studied, an NOEC of 6.3 mg a.i./L, 100 mg a.i./L and 100 mg a.i./L was established for Daphnia pulex, Thamnocephalus platyurus and Brachionus calyciflorus, respectively.

Conclusion: The acute 24-hour EC50 for Daphnia pulex, Thamnocephalus platyurus and Brachionus calyciflorus when exposed tro thiamethoxam under static conditions was determined to be > 100 mg a.i./L.

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments

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and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 26/ 1112004 Materials and Methods

Results and discussion Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 04 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors (Year) Knauer, K. (2000d)

Title 11 Acute toxicity test of CGA 293343 tech. to individual invertebrate species and molluscs from a natural pond assemblage under static conditions. Report No. / Date 2002642 / 21.07.2000 Syngenta File N° 293343/1273 Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Syngenta Crop Protection AG, Basel Switzerland 5. Dates of work 20th June 2000 through 5th July, 2000 6. Test substance ISO common name thiamethoxam; Company Code: CGA 293343 tech.,

Test System: Thiamethoxam technical, The acute toxicity of thiamethoxam (technical) to several invertebrate and mollusc species, notably Chaoborus Sp., a freshwater aquatic species, and Ostracoda, a freshwater crustacean, from the former group and Lymnea stagnalis and Radix peregra from the mollusc group of organisms. The test organisms were collected from a natural assemblage from the Novartis Crop Protection field aquatic ecosystem at Stein, Aargau, Switzerland, and transferred, together with the natural pond water, to an aquarium housed in the Novartis laboratories. The single species required for testing were accordingly separated and acclimatised for at least a period of 20-h. No feed was provided during the 48-h test period. Several range-finding tests were performed at various concentrations for the various species (0.001, 0.01, 0.1, 1, 10 and 100 mg/L nominal) prior to embarking upon the definitive study phase. Based on results from the range-finder tests, the definitive tests were conducted at the following nominal concentrations: Chaoborus Sp. Control (0), 1.6, 3.1, 6.3, 13, 25 and 50 mg thiamethoxam/L; Ostracoda Control (0), 0.016, 0.031, 0.063, 0.125, 0.25 and 0.50 mg thiamethoxam/L; Lymnea stagnalis and Radix peregra Control (0), 100 mg thiamethoxam/L, only. Analysis of fortified test solutions at 0-hr and 48-hrs indicated that measured exposure concentrations were in the range 82-102 % of the nominal concentrations for all test systems. Stock solutions and corresponding dilutions were performed in pond water. Four replicates per test concentration containing 5 organisms per replicate were prepared. Exposure of Chaoborus Sp. was performed in 250 mL glass test vessels containing 200 mL of test solution (thiamethoxam tech. dissolved

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in pond water). Petri dishes (60 mL) containing 40 mL of test solution were employed for the exposure of Ostracoda and both mollusc species. Test vessels containing the various species were incubated in a climatic chamber at 20 ± 2 °C for a period of 48-hours and subjected to a 16 hour light / 8 hour dark light regime, employing a 30 minute transition period. Average light intensity, during periods of illumination, was the order of 21 µE/m2sec during the Chaoborus Sp. and mollusc exposure tests, and 16 µE/m2sec during the test with Ostracoda. Water quality measurements included pH (range for all tests 7.4-9.2), temperature (constant at 20 °C), oxygen content (range for all tests 86-167% of saturation), water hardness and conductivity (334 mg CaCO3/L and 202 µS/cm, respectively). Immobilisation or other behavioural changes were recorded after 24- and 48-hours.

Acute toxicity of thiamethoxam (tech.) to Chaoborus Sp.

a Concentration Immobilisation Exposure EC50 [mg a.i./L] after 48 h period [mg a.i./L] [%] [hours] (95 % conf. interval) Nominal Measured Beginning / End control < 0.05 / < 0.05 0 24 h 6.9 1.6 1.51 / 1.43 0 3.1 3.06 / 2.97 5 6.3 5.99 / 6.14 80 48 h 5.5 13 13.3 / 12.7 90 25 24.4 / 23.0 100 50 46.1 / 45.0 100 NOEC^: 3.1 mg a.i./L a calculated based on nominal concentrations ^ Highest concentration with no immobilisation or behavioural changes

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Acute toxicity of thiamethoxam (tech.) to Ostracoda

a Concentration Immobilisation Exposure EC50 [µg a.i./L] after 48 h period [µg a.i./L] [%] [hours] (95 % conf. interval) Nominal Measured Beginning / End control < 2 / < 2 0 24 h 240 16 13.1 / 13.2 0 31 25.3 / 26.4 0 63 54.9 / 54.9 0 48 h 180 125 113 / 112 30 250 236 / 233 65 500 466 / 466 100 NOEC^: 63 µg a.i./L a calculated based on nominal concentrations ^ Highest concentration with no immobilisation or behavioural changes

Acute toxicity of thiamethoxam (tech.) to Lymnea stagnalis

a Concentration Immobilisation Exposure EC50 [mg a.i./L] after 48 h period [mg a.i./L] [%] [hours] (95 % conf. interval) Nominal Measured Beginning / End control < 0.05 / < 0.05 0 24 h > 100

100 94.8 / 98.8 0 48 h > 100

NOEC^: 100 mg a.i./L a calculated based on nominal concentrations ^ Highest concentration with no immobilisation or behavioural changes

Acute toxicity of thiamethoxam (tech.) to Radix peregra

a Concentration Immobilisation Exposure EC50 [mg a.i./L] after 48 h period [mg a.i./L] [%] [hours] (95 % conf. interval) Nominal Measured Beginning / End control < 0.05 / < 0.05 5 24 h > 100

100 93.9 / 92.7 10 48 h > 100

NOEC^: 100 mg a.i./L a calculated based on nominal concentrations ^ Highest concentration with ≤ 10% immobilisation or behavioural changes (NB - up to 10% immobilisation is considered acceptable in the control to fulfill test validity criteria)

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Conclusion: The 48-hour EC50 for the invertebrates Chaoborus Sp., Ostracoda when exposed to thiamethoxam under static conditions was 5.5 mg a.i./L and 0.18 mg a.i./L, respectively. The 48-hour EC50 was not attained for either of the two mollusc species tested, and thus is cited as > 100 mg a.i./L, where 100 mg a.i./L represents the highest test concentration studied. The NOEC was determined to be 3.1 mg a.i./L, 0.063 mg a.i./L for Chaoborus Sp. and Ostracoda, respectively, and 100 mg a.i./L for both Lymnea stagnalis and Radix peregra.

Results and discussion . Conclusion Reliability Acceptability Remarks COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 05 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors / Year Neumann, C. (1997a)

Title 12 ACUTE TOXICITY TEST OF CGA 322704 (METABOLITE OF CGA 293343) TO THE CLADOCERAN DAPHNIA MAGNA STRAUS UNDER STATIC CONDITIONS Report No. / Date 962528 / 31.01.1997 Novartis File N° Novartis Study # 322704-8 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection, Inc. Ecotoxicology Laboratories, CH-4002 Basle Switzerland 5. Dates of work August 27, 1996 through October 31, 1996 6. Test substance Company Code: CGA 322704,

7. Test method OECD Guideline No.: 202, Paris 1984 and FIFRA Guideline No.: 72-2, December 24, 1989. 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: CGA 3322704, The acute toxicity of CGA 322704 to Daphnia magna was determined in a 48-hour static test with five test substance concentrations and a blank control. At initiation, 120 daphnids were randomly selected and equally distributed among four replicates of each treatment and the control (5 animals/replicate). Each 150 mL test vessel contained 100 mL test solution or the blank control. The temperature during exposure was 20 ± 2°C. Five concentrations were employed of nominal 10, 18, 32, 58 and 100 mg/L. During exposure water concentrations were always close to nominal values both at test initiation as well as at the end of the end of exposure and were > 95% of nominals in all samples. Results were therefore expressed in nominal values. Findings: Acute toxicity of CGA 322704 (metabolite of thiamethoxam) to Daphnia magna

a Concentration Immobilisation Exposure EC50 [mg/L] after 48 h period [mg/L] [%] [hours] (95 % conf. interval)

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Nominal Measured End controls 0 0 10 9.6 0 24h > 100 18 17.3 0 48 h > 100 32 35.5 0 58 56.0 0 100 97.2 10 NOEC: 100 mg/L

• calculated based on nominal concentrations

Observations: After 48 hours of exposure, no immobilisation or behavioural changes of daphnids had occun ed at any of the test concentrations, nor in the controls.

Conclusion: The acute 48-hour EC50 for Daphnia magna when exposed to CGA 322704 metabolite of thiamethoxam under static conditions was detennined to be > 100 m IL. Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 30/ 11/2004 Materials and Methods

Results and discussion Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view of rapporteur member state Results and discussion Discuss ifdeviating f rom view ofrapporteur member state Conclusion Discuss ifdeviating f rom view ofrapporteur member state

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Reliability Discuss if deviating from view of rapporteur member state Acceptability Discuss if deviating from view of rapporteur member state Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 06 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors / Year Maetzler, P. (1998a)

Title 13 ACUTE TOXICITY OF CGA 355190 TO DAPHNIA MAGNA (IMMOBILISATION TEST) Report No. / Date G 54114, 982589 / 30.10.1998 Novartis File N° Novartis Study # 355190-3 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility Novartis Services AG, Scientific Services, Ecotox Center, CH-4002 Basle, Switzerland 5. Dates of work August 18, 1998 through August 20, 1998 6. Test substance Company Code: CGA 355190,

7. Test method OECD Guideline No.: 202, Paris 1984 and 92/69/EEC, C.2. 1992. 8. Deviations none 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: CGA 355190, The acute toxicity of CGA 355190 to Daphnia magna was determined in a 48-hour static test. The test was performed as a limit test with one test substance concentration (100 mg/L) and a blank control. For each concentration 20 Daphnia divided into two groups of 10 animals were used. Each 250 ml test vessel contained 60-100 mL test solution or the blank control. The temperature during exposure was between 19 and 20°C. During exposure water concentrations were always close to nominal values and were in the range of 99.5 at test initiation to 103.5% of nominals at the end of exposure in all samples. Results were therefore expressed in nominal values.

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Findings: Acute toxicity of CGA 355190 (metabolite of thiamethoxam) to Daphnia magna

Concentration Immobilisation Exposure EC so~ [mg/L] [mg/L] after 48 h period (95 % cont. interval) (%] (hours] Nominal Mean measured controls < 0.40 0 24h > 100 100 101.5 0 48 h > 100 NOEC: 100 mg/L

• calculated based on nominal concentrations

Observations: After 48 hours of exposure, no immobilisation or behavioural changes of daphnids had occuned at any of the test concentrations, nor in the controls.

Conclusion: The acute 48-hour EC50 for Daphnia magna when exposed to CGA 355190 (metabolite of thiamethoxam) under static conditions was detennined to be > 100 mg/L.

Results and discussion Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and M ethods Results and discussion

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Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 07 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors / Year Seyfried, B. (1998b)

Title 14 ACUTE TOXICITY OF NOA 407475 (METABOLITE OF CGA 293343) TO DAPHNIA MAGNA IN A 48-HOUR IMMOBILISATION TEST Report No. / Date 688803 / 22.09.1998 Novartis File N° Novartis Study # 407475-11 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility RCC Ltd, Environmental Chemistry & Pharmanalytics Division, Itingen, Switzerland 5. Dates of work July 06, 1998 through August 26, 1998 6. Test substance Company Code: NOA 407475,

7. Test method OECD Guideline No.: 202, Paris 1984 and 92/69/EEC, C.2. 1992. 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laboratory, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: NOA 407475, The acute toxicity of NOA 407475 to Daphnia magna was determined in a 48-hour static test. Five test concentration were employed with nominal values of 4.6, 10, 22, 46 and 100 mg/L. For each concentration 20 Daphnia divided into two groups of 10 animals were used. Each 100 mL test vessel contained 50 mL test solution or the blank control. Two test series with the same test concentrations but with different initial pH-values were performed. In the first series using freshly prepared, non- equilibrated test media, the initial pH-values ranged from 8.1 to 9.5 and were positively correlated with increasing concentration of NOA 407475 in the test media. In the second test series equilibrated test media with initial pH-values in the range of 8.2 to 8.4 were used. The temperature during exposure was 20°C ± 2. During exposure water concentrations were measured in the range from 86 to 109% of the nominal values. Results were therefore expressed in nominal values.

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Findings: Acute toxicity of NOA 407475 (metabolite ofthiamethoxam) to Daphnia magna

Concentration Immobilisation Exposure EC so* EC so .. (mg/L] after 48 h period pH not pH equilibrated equilibrated 1%1 fhoursl Imo /Ll rmo/Ll Nomin Mean pH not pH al measur equilibrat equlibrate ed ed d control - 0 0 s 4.6 - 0 0 24 h > 100 > 100 10 9.08 0 0 48h 91.5 82.9 22 19.4 0 15 46 39.6 0 25

100 87.5 5 55 48-h EC0 and NOEC: 10 mg /L • calculated based on nominal concentrations

Observations: After 48 hours of exposure, immobilisation of daphnids occun ed at concentrations of 22 and 46 mg/L for the not-equilibrated and the equilibrated test media, respectively. The slightly different 48h EC50 values found for the two test series can be explained by the usual biological va1iations in acute toxicity tests with Daphnia. They are unlikely to be due to the different pH-values at the start of the tests.

Conclusion: The acute 48-hour EC50 for Daphnia magna when exposed to NOA 407475 (metabolite ofthiamethoxam) under static conditions was detennined to be 91.5 mg/L (pH not equilibrated) and 82.9 mg/L (pH equilibrated).

Results and discussion

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Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 08 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors (Year) Wallace, S.J. (2002b)

Title 15 NOA 459602 (Thiamethoxam metabolite): Acute toxicity to daphnia magna Report No. / Date BL7244/B/ 19.04.2002 Syngenta File N° 459602/0017 Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Brixham Environmental Laboratory, Brixham, Devon, England 5. Dates of work 11th December 2001 through to 13th December 2001 6. Test substance Company Code: NOA 459602.,

7. Test method OECD - Guideline No.: 202, Part I, Paris 1984 8. Deviations None 9. GLP The study was conducted to conform with the UK Principles of Good Laboratory Practice (United Kingdom) GLP Regulations 1999) These principles are in accordance with the OECD Principles of Good Laboratory Practice, revised 1997 (ENV/MC/CHEM(98)17). These international standards are acceptable to the United States Environmental Protection Agency and this study, therefore satisfies the requirements of 40 CFR Part 160 and 40 CFR Part 792.

Test System: NOA 459602 (metabolite of thiamethoxam), . Test species: waterflea, Daphnia magna Straus; age: first instars less than 24 hours; source: in-house culture (Brixham Environmental Laboratory, England). Number of daphnids: for the test item test concentration and the water control four replicates each with 5 daphnids were used. Test vessels: 250 mL glass beakers containing 200 mL test medium (reconstituted water, Elendt’s M4). The test media were not exchanged and not aerated during the test. Nor were the daphnia fed during the test. All test vessels were placed under controlled environmental conditions at 21 ± 1 °C and a lighting regime of 16 hours light and 8 hours dark per day. The test concentration (nominal: 120 mg NOA 459602/L) was evaluated in this test. For the test concentration a single stock solution was prepared without using a solvent. In addition to the test item treatment a blank control with dilution water only was included. Test duration: 48 hours. The number of dead or immobilised organisms was counted at 24 and 48 hours. Dissolved oxygen and pH were measured at the start and end of the test. Total hardness of the water was determined at test start. Samples of the test media were taken from the excess of the freshly prepared test solutions and from the test vessels after 48 hours for the analytical determination (HPLC) of the test concentrations. Findings: Results are given on basis of nominal concentrations since concentrations did not vary being 100% of nominal at test start and 100 % of nominal at test end. The water temperature was 19.9-20.5 °C during the test period. The dissolved oxygen content ranged between 94.8% and 97.0% of saturation and pH values ranged from 7.97 to 8.14. The total hardness at test start was 220.3 mg CaCO3/L.

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Acute toxicity of NOA 459602 (metabolite of thiamethoxam) to Daphnia magna

a Immobilisatio Exposure LC50 Concentration n after 48 h period (95 % conf. [mg NOA 459602 / L] [%] [hours] interval) [mg NOA 459602 / L] Nominal Start End - Control <0.75 <0.75 0 120 120 120 0 48 h > 120 (n.a.) NOEC: 120 mg NOA 459602 / L a based on nominal concentrations; n.a. = not applicable

Observations: No immobilisation or abnormal behaviour was observed in either treatment during the test period.

Conclusion: The acute 48-hour LC50 for Daphnia magna exposed to NOA 459602 (metabolite of thiamethoxam) under static conditions was determined to be > 120 mg /L.

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 1/12/2004 Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks COMMENTS FROM ... Date Give date of comments submitted Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc lllA 7.4.1.2 Acute toxicity to invertebrates section No. 09 91 /414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Inve1·tebrates 2. Location in Dossier Section 6 3. Authors (Year) Knauer, K. (2000a)

Title 16 Acute Toxicity Test of CGA 293343 tech.to the ephemeroptera Cloen sp. tmder static conditions Report No. I Date 2002613 I 10.07.2000

Syngenta File N° 293343/1228

Source I Owner Unpublished I Syngenta Crop Protection AG

4. Testing facility Syngenta Crop Protection AG, Basel Switzerland 5. Dates of work 1st Jtme 2000 through 9th June, 2000 6. Test substance

7. Test method OECD-Guideline No.: 202, Paris 1984 EEC-C.2, December 29, 1992 U.S. EPA-FIFRA Guideline Number 72-2, December 24, 1989 U.S. EPA OPPTS Test Guideline OPPTS 850.1010, April, 1996 8. Deviations None 9. GLP GLP with the exception of range finding test, the study was conducted in compliance with the Siss Ordinance relating to Good Laborato1y Practice, adopted 2nd Febrna1y, 2000 (RS 813.016.5). This ordinance is based upon the OECD Principles of Good Laboratory Practice, as revised in 1997 and adopted 26th November, 1997 by decision of the OECD Council [C (97) 186/Final]. These procedm·es are based on the OECD Principles of Good Laborato1y Practice adopted 12th May, 1981 by the decision of the OECD Council [C (81) 30 (Final)] conceming Mutual Acceptance of Data in the Assessment of Chemicals, as ammended by the Cotmcil Decision-Recommendation of 2 October, 1989 conceming Compliance with Principles of Good Laboratory Practice Cotmcil [C (89) 87 (Final)].

Test System: Thiamethoxam technical, The acute toxicity of thiamethoxam (technical) to the aquatic insect Ephemeroptera Cloeon Sp. has been detennined at six different test concentrations and a blank control at a temperature of 19 °C. The test organism, Cloeon Sp. was collected from a natural assemblage from the Novartis Crop Protection field aquatic ecosystem at Stein, Aargau, Switzerland, then transfeITed, together with the natural pond water, to an aquarium housed in the Novaitis laboratories. The single species required for testing was accordingly sepai·ated. Cloeon Sp. were acclimatised for at least a period of20-h. No feed was provided during the 48-h test period. A range-finding test was performed at six concentrations (0. 001, 0. 01, 0 .1, 1, 10 and 100 mg/L nominal) prior to embarking upon the definitive study phase. Based on results from the range-finder, the definitive test was conducted at 0 (Control), 0.0031, 0.0063, 0.013, 0.025, 0.050 and 0.100 mg thiamethoxam/L. Analysis of fortified test solutions at 0-hr and 48-hrs indicated that measured exposure concentrations were, respectively, in the range 93-104% and 86-100% of the nominal concentrations. Four replicates per test concentration containing 5 organisms per replicate were prepared in 250 mL glass test vessels containing 200 mL of test solution (CGA 293343 tech. dissolved in pond water). Test vessels containing Cloeon Sp. were incubated in a climatic chamber for a period of 48-hours and subjected to a 16 hour light I 8 hour dark light regime, employing a 30 rninute transition period. Average light intensity, during periods of illumination, was in the order of20 ~1E/m2 sec. Water quality measurements included pH (range 8.2-8.5), temperature (constant at 19 °C) and oxygen content (87-96% of saturation) monitoring. Immobilisation or other behavioural changes were recorded after 24- and 48-hours.

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Findings: No immobilisation was observed in the control (blank pond water medium) dming the comse of the 48-hom exposme period, with no unexpected phenomena being observed. The test was thus deemed valid. The results are presented below .

Acute toxicity ofthiamethoxam (tech.) to Ephemeroptera Cloeon a Concentration Immobilisation Exposure EC so (µg a.i./L) after 48 h period [µg a.i./L) (%] (hours] (95 % cont. interval) Nominal Measured Beginning / End control < 0.5 I< 0.5 0 24h 19 3.1 3.05 I 3.11 0 6.3 6.55 I 6.32 0 13 11.9 I 11.2 50 48 h 14 25 25.0 / 24.2 90 50 48.6 I 47.8 100 100 92.5 I 91.1 100 NOEC A: 6.3 µg a.i./L

• calculated based on nominal concentrations " Highest concentration with no immobilisation or behavioural changes

Conclusion: The acute 48-houc EC50 for the Ephemeroptera Cloeon Sp. when exposed to thiamethoxam under static conditions was determined to be 14 µg a.i./L. The 48-houc NOEC was determined to be 6.3 µg a.i./L based on no evidence of immobilisation or behavioucal chan es. Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 2/12/2004 Materials and Methods

Results and discussion

Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods

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Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.2 / Acute toxicity to invertebrates section No. 10 91/414 Annex II Acute toxicity to invertebrates Point addressed 8.2.4

1. Annex point(s) II A, 8.2.4 Acute Toxicity to Aquatic Invertebrates 2. Location in Dossier Section 6 3. Authors (Year) Mank, M.A. and Krueger, H.O. (1998)

Title 17 CGA 293343 Technical: A 48-hour static acute toxicity test with the midge (Chironomus riparius). Report No. / Date 819-98 / 15.10.1998 Syngenta File N° 293343/0890 Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Wildlife International Ltd., Maryland, USA 5. Dates of work 29th September 1998 through 1st October 1998 6. Test substance ISO common name thiamethoxam; Company Code: CGA 293343 tech.,

7. Test method US-EPA, 1982, FIFRA Subdivision E, EPA 540/9-82-024. US-EPA, 1985, FIFRA Subdivision E, EPA 540/9-85-005. ASTM Standard Method E729-96a, 1994. 8. Deviations None 9. GLP The study was conducted in compliance with Good Laboratory Practice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989; OECD Principles of Good Laboratory Practice, OCDE/GD (92) 32, Environment Monograph No. 45, Paris 1992; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Bureau, 10 August 1984.

Test System: Thiamethoxam technical; Test organism: Chironomus riparius were obtained from a commercial supplier. Midges were cultured in water. Midge egg casings and newly hatched midges were held for six days (22 °C, pH 8.3-8.7, dissolved oxygen 8.4-8.6 mg/L) prior to collection for testing. Organisms used for the test were two days old and appeared to be in the 2nd instar development stage. Chironomus riparius were exposed to a geometric series of five test concentrations and a negative (dilution water - filtered well water decsribed as “moderately-hard”) Control. Two replicate test chambers (2 L test beakers containing 1 L of test solution, equivalent to a depth of 7.9 cm) were prepared for each treatment level, and 10 midges indiscriminately introduced into each test chamber. Nominal test concentrations were 0, 6.3, 13, 25, 50 and 100 µg a.i./L. Mean measured concentrations of between 100 and 105% of nominal were determined as the average of duplicate sample analyses taken at 0-hr and 48-hr. Test chambers were placed in a water bath and incubated at a constant temperature of 22 ± 1 °C during the course of the experiment. A photoperiod of 16 hours light (approximately 501 Lux) and 8 hours of darkness, applying a 30 minute transition period were used. Measured physical parameters included: water temperature, pH, dissolved O2, hardness, alkalinity and specific conductance. Observations were made periodically (after 1-, 24- and 48-hours) to determine the number of mortalities and immobility of test organisms, and the number of individuals exhibiting clinical signs of toxicity or abnormal behaviour were registered.

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Findings: Results from both the biological- and analytical-phase of the study are presented below. Acute toxicity ofthiamethoxam (tech.) to Chironomus riparius

Concentration Immobilisation Exposure ECso0 (µg a.i./L) after 48 h period [µg a.i./L) 1%1 lhoursl (95 % cont. interval\ Nominal Mean Measured Beginning I End control < LOQ /< 0 24 h 61 LOQ 6.3 6.88 I 6.40 0 13 13.3 I 12.4 0 48 h 35 25 26.8 / 25.2 10 50 53 .0 I 51.2 95 100 105 I 102 100 NOEC: 13 µg a.i./L • EC50 calculations based on mean meas1U'ed thiamethoxam concentrations Midges in the negative Control, 6.6 and 13 µg a.i./L treatment levels all appeared n01mal and healthy throughout the test. After an exposure period of 48-hr, mo1tality in the 26, 52 and 104 µg a.i./L treatment groups represented 10, 95 and 100 %, respectively.

Conclusion: The 48-hr EC50 and NOEC value for Chironomus riparius exposed to thiamethoxam fo1tified in the water-phase were determined to be 35 ~tg a.i./L and 13 ~tg a.i./L, respectively.

Results and discussion

Conclusion Reliability I Acceptability Remarks -

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COMMENTS FROM ... Date Give date of comments submitted Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc lllA 7.4.1.3 Growth inhibition test on algae section No. 01 91 /414 Annex II Effects on algal growth Point addressed 8.2.6 / 01

1. Annex point(s) II A, 8.2.6 E ffects on Algal Growth 2. Location in Dossier Section 6 3. Authors I Year Grade, R. (1996a)

Title 18 GROWTH INHIBITION TEST OF CGA 293343 TECH. TO GREEN ALGAE (SELENASTRUM CAPRICORNUTUM) IN A STATIC SYSTEM Report No. I Date 9500051 12.01.1996

Novartis File N° Novartis Study# CGA 293343-35 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection AG, Ecotoxicology Department, CH-4002 Basie Switzerland 5. Dates of work October 10, 1995 through October 25, 1995 6. Test substance ISO common name thiamethoxam.

7. Test method OECD - Guideline No. 201, Paris 1984, Guideline 92/69/EEC: C3. U.S. EPA Pesticide Assessment Guidelines, Subdivision J, Section No. 122-2 and 123-2 (1989). 8. Deviations None 9. GLP The study was conducted to conform with Good Laborato1y Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) OECD Principles of Good Laborato1y, May 1981. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: Thiamethoxam technical; After a preculture period of three days, Selenastru.m capricornutum imtla ce ens1ty 9700 cells/mL) was exposed to nominal test concentrations of 0.8, 1.6, 3.2, 6.4, 12.8, 25.6, 50 and 100 mg test substances/L for 72 hours. The measured test substance concenti·ations were 0.82, 1.59, 3.19, 6.12, 13.08, 26.75, 51.25, 100.45 at the strut of the study and 0.66, 0.93 , 1.92, 4.5, 9.91 , 20.56, 45.24, 81.78 mg test substance/L at the end of exposure, respectively. The toxicity values ai·e based on measured concenti·ations at the end of the study.

Findings:

Acute toxicity of thiamethoxam to green algae

Concentration Cell densities Inhibition .. 72-h EbCso u 72-h ErCsou (mg a.i./L] after 72 hours (growth rates) (95% cont. (95% cont. interval) interval)

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Nominal Measured End [cells x 10 [%) [mg a.i./L) [mg a.i./L) 4 /mLl I control 0 111 .9 0.0 0.8 0.66 126.7 0.0 1.6 0.93 120.3 0.0 > 81.8 > 81.8 3.2 1.92 143.3 0.0 none none 6.4 4.5 105.0 1.5

12.8 9.91 106.5 1.0

25.6 20.56 106.5 1.1 50 45.24 91 .5 5.8 NOErC: 81.8 mg a.i./L 100 81.78 109.8 0.3 • a negative inhibition indicates stimulation b calculated based on measm·ed end concentrations

Conclusion: The 72-hour ErC50 and EbC50 for Selenastrum capricornutum when exposed to thiamethoxam were detennined to be > 81.8 mg a.i./L.

Results and discussion

Conclusion Reliability Acceptability Remarks COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion

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Reliability Acceptability Remarks

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98/8 Doc lllA 7.4.1.3 Growth inhibition test on algae section No. 02 91 /414 Annex II Effects on algal growth Point addressed 8.2.6 I 03

1. Annex point(s) II A, 8.2.6 E ffects on Algal Growth 2. Location in Dossier Section 6 3. Authors I Year Grade, R. (1998a)

Title 19 GROWTH INHIBITION TEST OF CGA 293343 TECH. TO GREEN ALGAE (SELENASTRUM CAPRICORNUTUM) UNDER STATIC Report No. I Date CONDITIONS 972549 1 16.06.1998 Novartis File N° Novartis Study# CGA 293343-580 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection AG, Ecotoxicology Department, CH-4002 Basie Switzerland 5. Dates of work April 21 , 1997 through April 25, 1997 6. Test substance ISO common name thiamethoxam.

7. Test method OECD - Guideline No. 201 , Paris 1984 static system guideline 92/69/EEC: C3. U.S. EPA Pesticide Assessment Guidelines, Subdivision J, Section No. 122-2 and 123-2 (1982). EPA ecological test guidelines, OPPTS 850.54000, Public Draft, April 1996. Deviation of pH > 1.5 tmits 8. Deviations none 9. GLP The study was conducted to conform with Good Laborato1y Pra.ctice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) OECD Principles of Good Laborato1y, May 1981. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: Thiamethoxam technical; After a preculture period of three days, Selenastru.m capricornu.tu.m A so own as Pseudokirch. subcapitata SAG 61.81) (initial cell density 10100 cells/mL) was exposed to nominal test concentrations of 10, 18, 32, 58 and 100 mg thiamethoxam/L for 96 hours. The measured test substance concentrations were 10.1, 18.0, 32.8, 58.0 and 101.9 at the sta1t of the study and 6.8, 16.3, 29.7, 52.1 and 91.7 mg thiamethoxam/L at the end of exposure, respectively. The measured test concentrations of thiamethoxam were within 100 ± 20% of the nominal values. The toxicity values are, therefore, based on nominal concentrations.

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Findings:

Acute toxicity of thiamethoxam to green algae

Concentration Cell densities Inhibition .. 96-h EbCso u 96-h ErCsou (mg a.i./L] after 96 hours (growth rates) (95% cont. (95% cont. interval) interval) Nominal Measured End (cells x 10 (%] (mg a.i./L] (mg a.i./L] 4/mLl control 0 1081 . 10 6.8 1193 0.0 18 16.3 1376 0.0 > 100 > 100 32 29.7 1261 0.0 none none 58 52. 1 1165 0.0 100 91.7 111 2 0.0 NOErC: 100 mg a.i./L

• a negative inhibition indicates stimulation b calculated based on nominal concentrations

Conclusion: The 96-hour ErC50 and EbC50 for Selenastrum capricornutum when exposed to thiamethoxam were detennined to be > 100 mg a.i./L.

Results and discussion

Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and M ethods Results and discussion Conclusion Reliability

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Acceptability Remarks

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98/8 Doc IIIA 7.4.1.3 / Growth inhibition test on algae section No. 03 91/414 Annex II Effects on algal growth Point addressed 8.2.6 / 02

1. Annex point(s) II A, 8.2.6 Effects on Algal Growth 2. Location in Dossier Section 6 3. Authors / Year Grade, R. (1997)

Title 20 GROWTH INHIBITION TEST OF CGA 322704 (METABOLITE OF CGA 293343) TO GREEN ALGAE (SELENASTRUM CAPRICORNUTUM) UNDER STATIC CONDITIONS Report No. / Date 962529 / 09.01.1997 Novartis File N° Novartis Study # CGA 322704-7 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection AG, Ecotoxicology Department, CH-4002 Basle Switzerland 5. Dates of work November 12, 1996 through November 22, 1996 6. Test substance Company Code: CGA 322704,

7. Test method OECD - Guideline No. 201, Paris 1984, Gu ideline 92/69/EEC: C3. U.S. EPA Pesticide Assessment Guidelines, Subdivision J, Section No. 122-2 and 123-2 (1989). 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) OECD Principles of Good Laboratory, May 1981. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: CGA 322704, The influence of CGA 322704 on the growth of the green algal species Selenastrum capricornutum was investigated in a 72-hour static test. After a preculture period of three days, Selenastrum capricornutum (initial cell density 10200 cells/mL) was exposed to nominal test concentrations of 6.25, 12.5, 25, 50 and 100 mg CGA 322704/L for 72 hours. The measured test substance concentrations were 5.7, 11.0, 22.5, 44.0, 94.5 and 5.5, 10.1, 20.9, 45.5, and 91.2 mg CGA 322704/L at the start and the end of exposure, respectively. At all times measured start and measured end concentrations were close to nominal values, therefore, the toxicity values are based on nominal concentrations.

Findings:

Acute toxicity of CGA 322704 (metabolite of thiamethoxam) to green algae

b c c Concentration Cell densities Inhibition 72-h EbC50 72-h ErC50 [mg /L] a (growth (95 % conf. (95 % conf. after 72 hours rates) interval interval)

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Nominal Measured End [mean [%] [mg/L] [mg/L] emissionl I control 0 788.3 - 6.25 5.5 776.3 0.1 12.5 10. 1 766.3 0.3 > 100 > 100 25 20.9 731 .7 1.2 (107-1078) (216-37500) 50 45.5 679.7 2.6 100 91.2 497.7 8.5 NOErC: 50 mg/L

• mean emission: initial cell density (10200 cells/mL) = 3.88 b a negative inhibition indicates stimulation c calculated based on nominal concentrations

Conclusion: The 72-hour ErC50 and EbC50 for Selenastrum capricornutum when exposed to CGA 322704 (metabolite ofthiamethoxam) were determined to be > 100 mg IL.

Results and discussion

Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.3 / Growth inhibition test on algae section No. 04 91/414 Annex II Effects on algal growth Point addressed 8.2.6 / 04

1. Annex point(s) II A, 8.2.6 Effects on Algal Growth 2. Location in Dossier Section 6 3. Authors / Year Maetzler, P. (1998b)

Title 21 TOXICITY OF CGA 355190 TO GREEN ALGAE (GROWTH INHIBITION TEST) Report No. / Date

G54117, 982590 / 30.10.1998 Novartis File N° Novartis Study # 355190-4 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility Novartis Services AG, Sientific Services, Ecotox Center, CH-4002 Basle Switzerland 5. Dates of work September 08, 1998 through September 11, 1998 6. Test substance Company Code: CGA 355190,

7. Test method OECD - Guideline No. 201, Paris 1984, Gu ideline 92/69/EEC: C3. 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) OECD Principles of Good Laboratory, May 1981. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: CGA 355190, The influence of CGA 355190 on the growth of the green algal species Pseudokirchneriella subcapitata alias Scenedesmus subspicatus was investigated in a 72-hour static test. After a preculture period of three days, the green algae (initial cell density 10,000 cells/mL) were exposed to nominal test concentrations of 4.3, 9.4, 21, 45 and 100 mg CGA 355190/L for 72 hours. The measured test substance concentrations were in the range of 98.8 - 100.5% of nominal values during the test. At all times measured start and measured end concentrations were close to nominal values, therefore, the toxicity values are based on nominal concentrations.

Findings:

Acute toxicity of CGA 355190 (metabolite of thiamethoxam) to green algae

b c c Concentration Cell densities Inhibition 72-h EbC50 72-h ErC50 [mg/L] a (growth (95 % conf. (95 % conf. after 72 hours rates) interval interval)

Nominal Measured End [biomass] [%] [mg/L] [mg/L] control 0 712.44 -

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4.3 n.d. 728.78 -0.7 9.4 n.d. 695.44 0.6 > 100 > 100 21.0 n.d. 699.44 0.5 (-) (-) 45.0 43.9 730.56 -0.7 100 99.0 621.78 3.1 NOErC: 100 mg/L • pa1t1cle number x 1000 b a negative inhibition indicates stimulation c calculated based on nominal concentrations

Conclusion: The 72-hour ErC50 and EbC50 for Selenastn11n capricornutum when exposed to CGA 355190 (metabolite ofthiamethoxam) were determined to be > 100 mg IL.

Results and discussion

Conclusion Reliability I Acceptability Remarks -

COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc IIIA 7.4.1.3 / Growth inhibition test on algae section No. 05 91/414 Annex II Effects on algal growth Point addressed 8.2.6 / 04

1. Annex point(s) II A, 8.2.6 Effects on Algal Growth 2. Location in Dossier Section 6 3. Authors / Year Seyfried, B. (1998c)

Title 22 TOXICITY OF NOA 407475 (METABOLITE OF CGA 293343) TO SENEDESMUS SUBSPICATUS IN A 72-HOUR ALGAL GROWTH INHIBITION TEST Report No. / Date

688825 / 14.08.1998 Novartis File N° Novartis Study # 407475-9 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility RCC Ltd., Environmental Chemistry & Pharmanalytics Division, Itingen, Switzerland 5. Dates of work July 03, 1998 through July 22, 1998 6. Test substance Company Code: NOA 407475,

Test System: NOA 407474, The influence of NOA 407475 on the growth of the green algal species Scenedesmus subspicatus was investigated in a 72-hour static test. After a preculture period of three days, Scenedesmus subspicatus (initial cell density 10,000 cells/mL) was exposed to nominal test concentrations of 1.0, 2.2, 4.6, 10.0, 22.0 and 46.0 mg/L. Additionally, a control was tested in parallel. The mean analytically determined concentrations of NOA 407475 in the analyzed test media ranged from 86 to 101% of the nominal values during the test period of 72 hours. At all times measured start and measured end concentrations were close to nominal values, therefore, the toxicity values are based on nominal concentrations.

Findings:

Acute toxicity of NOA 407475 (metabolite of thiamethoxam) to green algae

b c c Concentration Cell densities Inhibition 72-h EbC50 72-h ErC50 [mg/L] a (growth (95 % conf. (95 % conf. after 72 hours rates) interval interval)

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Nominal Mean (cells x 10 (%] (mg/L] (mg/L] 4 Measured /mLl I control - 9 1.6 0.0 1.0 - 97.0 -1.7 2.2 - 90.5 0.0 14.0 33.8 4.6 4.6 87.6 0.6 (6.9-20.3) (20.5-125.4) 10.0 9.1 41.2 17.3

22.0 19.0 19.7 33.7 NOErC and NOEbC: 4.6 mg /L

46.0 40 6.1 60.0

• initial cell density (10'000 cells/mL) b a negative inhibition indicates stimulation c calculated based on nominal concentrations

Conclusion: The 72-hour ErC50 and EbC50 for Selenastrum subspicatus when exposed to NOA 407475 (metabolite ofthiamethoxam) were dete1mined to be 33.8 mg IL and 14.0 mg /L, respectively.

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 14/12/2004 Materials and Methods Results and discussion Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.1.3 / Growth inhibition test on algae section No. 06 91/414 Annex II Effects on algal growth Point addressed 8.2.6

1. Annex point(s) II A, 8.2.6 Effects on Algal Growth 2. Location in Dossier Section 6 3. Authors (Year) Wallace, S.J. (2002c)

Title 23 NOA 459602 (Thiamethoxam metabolite): Acute toxicity to the green algae (Selenastrum capricornutum) Report No. / Date BL7245/B/ 25.04.2002 Syngenta File N° Syngenta Study # 459602-0018 Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Brixham Environmental Laboratory, Brixham, Devon, England 5. Dates of work 10th December 2001 through to 14th December 2001 6. Test substance Company Code: NOA 459602.,

7. Test method US Environmental Protection Agency, Office of Prevention, Pesticides and Toxic Substances, Ecological Effects Test Guideline OPPTS 850.5400 Algal Toxicity, Tiers I and II “Public Draft”. April 1996 8. Deviations None 9. GLP The study was conducted to conform with the UK Principles of Good Laboratory Practice (United Kingdom) GLP Regulations 1999) These principles are in accordance with the OECD Principles of Good Laboratory Practice, revised 1997 (ENV/MC/CHEM(98)17). These international standards are acceptable to the United States Environmental Protection Agency and this study, therefore satisfies the requirements of 40 CFR Part 160 and 40 CFR Part 792.

Test System: NOA 459602 (metabolite of thiamethoxam), Freshwater green algae, Selenastrum capricornutum (strain: ATCC 22662; from an exponentially growing 3 day-old pre-culture; source: in-house culture) were exposed to five concentrations of the test item under static conditions for a period of 96 hours. The following nominal test item concentrations were tested: 7.5, 15, 30, 60 and 120 mg NOA 459602 per litre. A single stock solution (the highest test concentration) was prepared by mixing an appropriate amount of the test item directly into the sterile culture medium without additional solvent. Dilution of aliquots of this stock solution with sterile culture medium yielded the lower concentrations. A blank control with test medium only was included. The starting concentration of algal cells was approximately 1.01 x 104 cells/mL. Three replicate test chambers were maintained for the test item treatment groups and six replicates for the control group. The test flasks comprised 250 ml conical glass flasks each containing 100 mL of the appropriate test solution/algal suspension. The flasks were randomly positioned within a laboratory shaker and continuously shaken at 100 rpm under continuous illumination. Samples from each replicate of the test groups were collected at approximately 24-hour intervals for the determination of the algal cell density by means of a cell counter. The temperature of the incubator was measured daily. The pH was recorded at 0 h and 96 h. Test item concentrations were measured (HPLC) on samples taken from the excess test solutions at test start and from the flask at the end of the test.

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Findings: Results are given on basis of nominal concentrations since concentrations va1ied between 100 to 11 3% of nominal at test strut and 107 to 113% of nominal at test end. Dming the test pe1iod, temperature was maintained at 23 .9-24.0 °C and the light intensity measured once during the study was 4040 lux (49.1 µE/m2·sec, in te1m s of quantum response). The pH ranged from 7.41 to 7.47 at test start, and 9.28 to 9.93 at test te1mination (increase in pH due to the massive growth of algae in control and test item groups).

Acute toxicity of NOA 459602 (metabolite ofthiamethoxam) to green algae

Concentration Cell density nhibition (AUC 96-h ErCso a 96-h EbCso a Ima NOA 459602 I L1 after 96 hours • 96 h (95% confid. limit) (95% confid. limit) Nomina Measured (cells x 1O 4/mL] (%] (mg NOA 459602 / L] (mg NOA 459602 I L] I DayO Day 4 Contro <0.75 <0.75 388 - 1 7.5 8.0 8.2 390 1 > 120 (n.a.) > 120 (n.a.) 15 17 17 378 2 30 31 33 369 6 60 61 64 348 10 NOErC : NOEbC: 120 120 130 314 17· 60 mg NOA 459602/ 60 mg NOA 459602/ L L based on nominal concentrations n.a. not applicable *significant difference (P=0.05) from the culture meditun

Observations: The density of the cells in the control group increased by factor 388 during the course of the test (exponential growth), demonstrating the validity of the test system. In the test item treatment after 96 hours a dose-responsive inhibition was obse1ved. However, at all test item concentrations tested the percentage inhibition compared to the controls was < 50% and only in the highest test item concentration was this inhibition > 10% and statistically significant.

Conclusion: The acute EbCso (96 h) for green algae exposed to NOA 459602 (metabolite of thiamethoxam) was dete1mined to be > 120 mg IL.

Results and discussion Conclusion Reliability Acceptability

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Remarks COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc lllA 7.4.1.4 Inhibition of microbiological activity section No. 01 91 /414 Annex II Effects on biological methods for sewage treatment Point addressed 8.7 I 01

1. Annex point(s) II A, 8.7 Effects on biological methods for sewage treatment 2. Location in Dossier Section 6, 3. Authors (year) Grade, R. (1996b) Title Repo1t on the test for activated sludge respiration inhibition of CGA 293343 tech.

Report No., Date 950002, 8 Janua1y 1996 Novartis File N° (Desire) 293343/34 Owner Novartis Crop Protection AG 4. Testing facility Ciba-Geigy Ltd. Ecotoxicology Department CH-4002 Basie Switzerland 5. Dates of work Study Initiation: 4 September 1995 Experimental Start: 20 September 1995 Experimental Termination: 20 September 1995 Study Completion: 4 September 1995 6. Test substance ISO common name thiamethoxam.

7. Test method OECD- Guideline No.: 209 (Paris 1984) A settled sludge was used instead of a centrifuged sludge. The test substance was directly applied to the test medium and not as a stock solution. 8. Deviations none 9. GLP Yes , with the exception of the deviations to the protocol, the study was pe1fon11ed in compliance with Good Laborato1y Practice (OLP) in Switzerland, Procedm·es and Principles, March 1986 (Ve1fahren und Gmndsiitze der Outen Labo1praxis (OLP) in der Schweiz, Miirz 1986), issued by the Swiss Federal Department of the Interior and the Intercantonal Office for the Control of Medicaments, Switzerland. These procedures are based on the OECD Principles of Good Laboratory Practice, adopted May, 1981 by the decision of the OECD Council [C (81) 30 (Final)] concerning mutual acceptance of data in the assessment of chemicals.

Test System: CGA 293343 technical; . Activated sludge from the Reinach Sewage Treatment P ant m Switzer an was e a standard amount of synthetic sewage. The pH of the sludge before use was 8.4. After settlement, 1.72 g sludge/L inoculate were treated with five test doses of thiamethoxam ranging from 1.0 to 100.0 mg a.i./L, two treatment blanks and three doses of a toxic standard (3 ,5-dichlorphenol) ranging from 3.2 to 32.0 mg a.i./L. Nominal concentrations were used. Oxygen consumption of bacteria was measured for three hours.

Findings:

Effects ofthiamethoxam on activated sludge respiration

Nominal cone. Consumption Inhibition • EC so (mg a.i./L] rate (%] (mg a.i./L] rma 02/l/hl 1.0 69.2 1 3.2 64.9 7

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10.0 72.7 -4 32.0 50.0 28 100.0 77.8 -11 > 100 Blank 1 70.6 -1 Blank 2 69.2 1 toxic standard 3.2 64.9 7 10. 53.3 24 32.0 15.6 78 2.19 a negative inhibition indicates stimulation

Conclusion: The 3-hour EC20, EC50 and EC80 values were estimated to be >100 mg/L. No dose response relationship between the test substance doses and the Oxygen consumption rates were found, implying that CGA 293343 did not have an inhibito1y effect on activated sludge respiration.

Results and discussion

Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion

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Conclusion Reliability Acceptability Remarks

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98/8 Doc lllA 7.4.3.1 Prolonged toxicity to an appropriate species of fish section No. 01 91 /414 Annex II Chronic toxicity test on juvenile fish Point addressed 8.2.2.1 01

1. Annex point(s) II A, 8.2.2.1 Chronic Toxicity Test on Juvenile Fish 2. Location in Dossier Section 6 3. Authors I Year

Title 24 PROLONGED TOXICITY TEST OF CGA 293343 TECH. TO RAINBOW TROUT (ONCORHYNCH US M YKISS) IN THE FLOW-THROUGH Report No. I Date SYSTEM 95R003/ 30.07.1997 Novartis File N° Novartis Study # 293343-296 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility

5. Dates of work Febiua1y 5, 1997 through March 25, 1997 6. Test substance ISO common name thiamethoxam.

7. Test method OECD Guideline No.: 204, Fish- Prolonged Toxicity Test, 04/04/ 1984 8. Deviations None 9. GLP The study was conducted to conform with Good Laborato1y Pra.ctice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Envirorunental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) OECD Principles of Good Laborato1y, May 1981. 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Blllea, IO August 1984.

Test System: Thiamethoxam technical; In a prolonged exposure test, effects of thiamethoxam on t e growt an e av10ur o JUVem e Rainbow trout (Oncorhynchus mykiss) were investigated under flow-through conditions. The crite1ia for effects were m01tality, weight gain, increase in length, food conversion index and other non-lethal effects on behaviour. Survival and growth were monitored dming a pe1iod of28 days of exposure. The study was performed with one tank per concentration and control containing ten fish each. Fish were individually marked and had a mean body weight of 1.8 g and a mean length of 57 mm at the beginning of the test. Five concentrations between 10 and 100 mg a.i./L were tested. The test substance appeared homogeneously distributed in the test vessels at all test times and test concentrations. The actual mean concentrations of thiamethoxam were 9.4, 19, 34, 60, and 109 mg a.i./L. con esponding to 95-109% of the nominal concentrations. Results are based on nominal concentrations.

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Findings: Prolonged toxicity of thiamethoxam to Rainbow trout Concentration Mortality Food Mean length of Mean wet weight [mg a.i./L] conversion juveniles (28d) of juveniles (28d) Index Nominal Mean [%] [mm] [mg] measured Control 0 0 112.3 74.0 4.2 10 9.5 0 143.3 72.3 3.8 18 19 0 116.4 74.0 4.0 32 34 0 113.5 73.2 3.8 58 60 0 115.7 70.9 3.6 100 109 0 137.0 72.7 3.8

Observations: After 28 d exposure, no sublethal effects such as growth parameters (rate of weight gain, change in length and food conversion), feeding activity, change in swimming behaviour or respiratory function, pigmentation, loss of equilibrium as well as a reaction to external stimuli were observed at any concentration. Conclusion: The NOEC (28 days) with regard to lethal and sublethal effects of thiamethoxam on Rainbow trout was 100 mg a.i./L. The LOEC was > 100 mg a.i./L.

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 15/12/2004 Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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Section A 7.4.3.(01) Effects on aquatic organisms - further studies Annex Point IIIA XIII.3

Official 1 REFERENCE use only 1.1 Reference Ashwell, J., Dark, R. Emburey, S. (2003) Thiamethoxam 25 WG (A9584C) Outdoor Microcosm Study to Assess Effects on Aquatic Organisms. Syngenta, Jealott's Hill International Research Centre, Bracknell, Berkshire, UK. Unpublished Repo1t no. RJ3379B (Syngenta File no. CGA293343/1851). Study dates. 15th April - 19th September 2002. 1.2 Data protection Yes. 1.2.1 Data owner Syngenta Crop Protection. 1.2.2 Companies with letter of access .. 1.2 .3 Criteria for data protection

2 GUIDELINES AND QUALITY ASSURANCE 2.1 Guideline study CLASSIC (Community Level Aquatic System Studies - Interpretation Criteria)(2002). Proceedings from the 1999 CLASSIC workshop. Society of Environmental Toxicology and Chemistiy (SETAC), Florida and SETAC -Europe Brussels. HARAP (1999). Guidance document on higher-tier aquatic risk assessment for pesticides. Society of Environmental Toxicology and Chemistiy-Europe, Bmssels 179pp. 2.2 GLP Yes. 2.3 Deviations No.

3 MATERIALS AND METHODS 3.1 Test material 3.1.1 Lot/Batch number 3.1.2 Purity 3.2 Refer ence Not used. substance 3.3 Testing procedure An outdoor pond microcosm study was conducted to investigate the effects of A-9584 C on conununities of freshwater organisms. The test systems were rectangular varied depth microcosms containing approximately 1300 L of water over 10 cm sediment, stocked with macrophytes and inve1tebrates. Cormnunities were allowed to establish in the inicrocosms for a period of 12 weeks before chemical application. The replicated study design consisted of five A-9584 C treatinents at 100, 30, 10, 3 and 1 µg ai/L, and an untreated conti·ol. A single application of A-9584 C was made to the microcosms on 18th June 2002. Application was made by direct addition and thorough mixing to rapidly disti-ibute the fonnulation throughout the water column - taking the toxicological approach favoured by HARAP (Guidance document on Higher Tier Aquatic Risk Assessment for Pesticides, 1999) and CLASSIC (Community-Level Aquatic System Studies-Interpretation Criteria, 2002 . Ph sico-chemical ro e1ties of the water and

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Section A 7.4.3.(01) Effects on aquatic organisms - further studies Annex Point IIIA XIII.3 phytoplankton, zooplankton and macroinvertebrate communities were studied for up to 93 days after application. Multivariate analysis (Principal Response Curves method) was used to identify any treatment-related changes in community composition. Analysis of variance was also used to look at population level differences for selected taxa; those not analysed were considered to be either too variable in occw1·ence or not abundant enough to permit meaningful conclusions to be drawn.

4 RESULTS

4.1 Range finding test Not perfo1med. 4.2 Results test substance 4.2.l Initial concentration Initial concentrations of thiamethoxam in the microcosms were of test substance measmed at betv.•een 81and130 % of nominal (mean 99 %), and so it was considered that the appropriate treatment levels had been achieved at dosing. 4.2.2 Effect data No long-te1m ecologically adverse effects (on physicochemical parameters or communities of phytoplankton, zooplankton and macroinve1tebrates) were observed at any of the treatment levels (up to and including 100 µg ai/L). An effect on Chironomidae was observed at 100 µg ai/L; however, this was an isolated event seen only in the emergence trap samples on day 15, and was not observed at any subsequent sampling points. The rapid dissipation of thiamethoxam in the microcosms as compared to laboratory-based tests is the likely explanation for effects seen in laboratory tests not being observed in this study.

5 APPLICANT'S SUMMARY AND CONCLUSION 5.1 Materials and As above. methods 5.2 Results and Initial concentrations of thiamethoxam in the microcosms were discussion measmed at betv.re.en 81and130 % of nominal (mean 99 %), and so it was considered that the appropriate treatment levels had been achieved at dosing. Thiamethoxam concentrations then declined rapidly following application with treatment means ranging from 34 % to 60 % of applied after 3 days. Concentrations were at or below 0.6 µg/L in all microcosms after 21 days. Results are reported in relation to nominal application concentrations of thiamethoxam. 5.2.1 NOEC 30 µg ai/L. 5.3 Conclusion The No Observed Ecologically Adverse Effect Concentration (NOEAEC) of A-9584 C in an outdoor pond microcosm study was 100 µg ai/L, and 30 µg ai/L was determined to be the overall NOECcommunity 5.3.l Reliability 1 5.3.2 Deficiencies No

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Section A 7.4.3.(01) Effects on aquatic organisms - further studies Annex Point IIIA XIII.3 Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted

EVALUATION BY RAPPORTEUR MEMBER STATE Date 7-03-06 Materials and Methods Results and discussion Conclusion Reliability Acceptability

Remarks

COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc lllA 7.4.3.2 Effects on reproduction and growth rate on an appropriate species section No. 01 offish 91 /414 Annex II Fish early life stage toxicity test Point addressed 8.2.2.2 01

1. Annex point(s) II A, 8.2.2.2 Fish early life stage toxicity test 2. Location in Dossier Section 6 3. Authors I Year

Title 6 CGA-293343: AN EARLY LIFE-STAGE TOXICITY TEST WITH THE RAINBOW TROUT (ONCORHYNCHUS MYKISS) Report No. I Date 322-96, 108A-188 / 14.02.1997

Novartis File N° Novartis Study# 293343-205 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility Wildlife International Ltd, 8598 Collllllerce Drive, Easton, MD 21601 5. Dates of work September 13, 1996 through December 12, 1996 6. Test substance ISO common name thiamethoxam.

7. Test method U.S. EPA Pesticide Assessment Guidelines, Subdivision E, Section No. 72-4(a) (1982), ASTM Standard E 1241-88 (1988) Standard Guide for conducting Early Life-Stage Toxicity Tests with Fish, and US EPA Standard Evaluation Procedure , Fish Early Stage Test. 8. Deviations none 9. GLP The study was conducted to conform with Good Laborato1y Pra.ctice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989; OECD, ISBN 92-84-12367-9, Paris 1982; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Bm·ea, 10 August 1984 ..

Test System: Thiamethoxam technical; In a sub-chronic test, effects of thiamet oxam on ear y 1 e stages o Ram ow trout (Oncorhynchus mykiss) were investigated under flow-through conditions. The criteria for effects were hatching success of embryos, time to hatch, time for larvae to swim-up and growth and smvival of juveniles during a study period of 88 days. Fish embryos were exposed to a geometric series of five concentrations and a negative control. Four replicate test chambers were maintained in each treatment and control group, with each test chamber containing two incubation cups. The test was initiated with the distribution of newly-fertilised eggs to the incubation cups. Each cup contained 15 embryos resulting in a total of 30 embryos per replicate and 120 embryos per concentr·ation group. The total exposure period was 88 days which included a 28-day hatching period and a 60-day post-hatch period. Insoluble material was not observed and the substance appeared in solution in all test vessels and at all times and concentrations. The mean measured concentr·ations of thiamethoxam based on weekly measurements were, 1.3, 2.5, 5.1 , 10 and 20 mg test substance/Lin the tr·eated groups. The following calculations and values ar·e based on mean measured concentrations.

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Findings: Toxicity of thiamethoxam to the early life stages of Rainbow trout Concentration Percent Percent survival Mean length of Mean wet weight of (mg a.i./L] hatch 18 d post hatch juveniles 60 d post juveniles 60-d post hatch hatch Nominal Mean (%] (%] (mm] (mg] measured control 0 77 98 50.5 0.277 1.3 1.3 73 99 51.3 0.28 2.5 2.5 74 99 51.1 0.277 5.0 5.1 75 100 50.9 0.277 10 10 76 100 51.3 0.283 20 20 73 100 51.7 0.29

Observations: There were no apparent treatment-related effects on time to hatch, hatching success, time to reach swim-up stage of development, la1vae smvival, fry smvival or growth of Rainbow trout exposed to thiamethoxam. Conclusion: The NOEC was 20 mg a.i./L, the highest concentration tested, while the LOEC were estimated to be greater than 20 mg a.i./L.

Conclusion Reliability I Acceptability Remarks -

COMMENTS FROM ... Date Materials and M ethods Results and discussion Conclusion

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Reliability Acceptability Remarks

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98/8 Doc lllA 7.4.3.4 Effects on reproduction and growth rate with an appropriate invertebrate section No. 01 species 91 /414 Annex II Chronic toxicity to aquatic invertebrates Point addressed 8.2.5 / 01

1. Annex point(s) II A, 8.2.S Chronic Toxicity to Aquatic Inve1·tebrates 2. Location in Dossier Section 6 3. Authors I Year Neumann, C. (1997b)

Title 7 DAPHNIA MAGNA REPRODUCTION TEST: EFFECTS OF CGA 293343 ON THE REPRODUCTION OF THE CLADOCERAN DAPHNIA MAGNA STRAUS IN A SEMI-STATIC LABORATORY TEST Report No. I Date 95G004 / 24.09.1997 Novartis File N° Novartis Study# 293343-323 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection, Inc. Ecological Toxicology Laborato1y, CH-4002, Basie, Switzerland. 5. Dates of work September 25, 1996 through Febmaiy 26, 1997. 6. Test substance ISO common name thiamethoxam.

7. Test method OECD Guideline No.: 202, Paris 1984 (revised draft ofOECD Guideline 202 Pait II, Janua1y 1996), FIFRA Guideline No.72-4, December 24, 1989. 8. Deviations none 9. GLP The study was conducted to conform with Good Laborato1y Pra.ctice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989 1) OECD Principles of Good Laborato1y , May 1981. 4) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Bmea, 10 August 1984.

Test System: Thiamethoxam technical; Test organism: Daphnia magna; Age: ~ 24 hours old; Daphnids were exposed to a geometiic series of five concenti·ations of the test substance and a water conti·ol under semi-static conditions with renewal of the test solution three times a week. Ten replicate test chambers were maintained in each treatment and the conti·ol group. Nominal test concenti·ations were: 6.0, 12.5, 25.0, 50.0 and 100 mg/L. Measured contents of thiamethoxam in all test concentrations were greater than or equal to 90% of the nominal values throughout the exposure pe1iod with two exceptions of 89.6 and 81.6% of nominal for 12.5 mg/L on days 5 and 19 of exposure. Measured end concentrations were 6.5, 12.5, 24.9, 50.2, and 100.3 100 mg/L. The results were repo1ted based on nominal values. Biological obse1vations on adult smvival, immobilisation and changes in behaviour or appearance were recorded daily. With the onset of brood production, young smvival and immobilisation were also recorded daily.

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Findings: Reproductive toxicity of thiamethoxam to Daphnia magna Concentration Adult Mean Time Immobilisation Mean Mean Adult (mg a.i./L] mortality to first brood of young Young/ Length daphnids female Measured 21 d 21 d 21 d Nominal end (number] (days] (%] (number] (mm] Control 0 0 9.0 0 122.2 4466 6 6.5 1 9.6 0 105.9 4312 12.5 12.5 1 8.9 0 117.3 4177 25 24.9 0 9.2 0 119.7 4432 50 50.2 1 9.0 0.8 125.4 4406 100 100.3 1 9.1 0 101.9 4282

Observations: Parental viability as well as the reproductive output of the cladoceran Daphnia magna strauss were not affected by the treatment with thiamethoxam up to nominal concentrations of 100 mg/L. Nor were clinical signs of toxicity or any other sublethal effects observed among the parent and the off-spring generation. Conclusion: The No-observed-effect concentration was determined to be 100 mg/L (NOEC reproduction 2l d). The lowest-observed-effect concentration (LOEC reproduction 2ld) for the number of young produced per parent animal was not found within the test concentration range and was estimated to be greater than 100 mg/L.

Results and discussion

Conclusion Reliability I

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98/8 Doc lllA 7.4.3.5.1 Effects on sediment dwelling organism section No. I 01 91 /414 Annex II Effects on sediment dwelling organisms Point addressed 8.2.7 / 01

1. Annex point(s) II A, 8.2.7 Effects on sediment dwelling organisms 2. Location in Dossier Section 6 3. Authors I Year Grade, R. (1998b)

Title 8 TOXICITY TEST OF CGA 293343 TECH. ON SEDIMENT-DWELLING CHIRONOMUS RIPARIUS (SYN. CHIRONOMUS THUMM[) UNDER STATIC CONDITIONS Report No. I Date\ 972552 / 02.10.1998 Novartis File N° Novartis Study# 293343-720 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection, Inc., Ecotoxicology Laboratories, CH-4002, Basie, Switzerland 5. Dates of work Febiua1y 24, 1998 through May 5, 1998 6. Test substance ISO common name thiamethoxam.

7. Test method BBA Guideline Proposal, 1995, Effects of Plant Protection Products on Sediment-Dwelling Larvae of Chironomus riparius in a water-Sediment System. OECD Guideline for Testing of Chemicals, Proposal for toxicity Test with Chironomidae, November 1997. 8. Deviations None 9. GLP The study was conducted to conform with Good Laborato1y Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989 1) OECD Principles of Good Laborato1y, May 1981. 4) Japan MAFF, 59 NohSan, Notification No. 3850, Agriculttu·al Production Bm·ea, 10 August 1984.

Test System: Thiamethoxam technical; ;. Test organism: Chironomus riparius larvae (syn. Chironomjus t ummi ; Age: 2-3 ys o instar larvae. Two exposure scenarfos were included in a water-sediment test system. In scenarfo A, thiamethoxam was applied to the water phase simulating a spray drift event. In scenarfo B, which was intended to simulate a nm-off event, the compound was added to the sediment. The tests were perfo1med in 1-L glass beakers containing approximately 1.6 cm sediment and a water column of 8 cm height at the beginning and of about 6 cm height at the end of the study. A constant temperature of20 ± 2°C was maintained during the test with a photoperiod of 16 hours light and 8 hours dark Exposure Scenruio A: A range of concentrations ofthiamethoxam (1.25, 2.5, 5, 10, 20 and 50 µg/L) was applied below the smface to the water column of sediment-water systems using a pipette. The test was canied out under static conditions. Twenty-four hours before addition of the test substance 20 1st instar la1vae of Chironumus riparius were introduced in the test vessels.

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Measured test concentrations in the water phase were below the limit of detection for the first four concentrations and ranged between 9 and 33 µg a.i./L for the three highest concentrations at 1 to 3 hours after application of the substance. At the end of the study (day 30) measured concentrations had decreased below the limit of detection at all test concentrations. None of the metabolites could be identified. Results are based on nominal concentrations in the water phase. Exposure Scenario B: Thiamethoxam treated sand was mixed with aged artificial sediment at a range of concentration (12.5, 25, 50, 100, 200 and 400 µg/kg sediment dry weight). The spiked sediment and water were added to the test vessels 21 hours prior to the introduction of Chironomus larvae and the start of the test. Actual measured test concentrations in the water phase were below the limit of detection at test day 0 after addition of the larvae, except for the two highest dose levels (measured concentrations 6 and 11 µg/L in water). At the end of the study corresponding values were still below the limit of detection and were 11 and 13 µg/L for the two highest dose levels. In the sediment, measured concentrations ranged from being below the limit of detection to 160 µg/ kg sediment for the highest dose level (400 µg/kg sediment nominal) at test day 0. For this dose level, measured concentrations were 86, 76 and 0% of nominal values at day 0, 7 and 30, respectively. Traces of CGA 355190 were detected in these samples at day 30. No other metabolites could be identified in the remaining samples from any concentration level. Results were based on nominal concentrations in the sediment. The biological assessment was based on impacts on full maturation of the larvae to adult midges. Main parameters examined were the rate and time of emergence and the total number of emerging male and female midges.

Findings:

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Scenario A: Application to the Water Phase

Table 7.4.3.5.1/01-1 Chronic toxicity of thiamethoxam to Chironomus riparius following application to the water phase

a a Concentration Emergence Development 30-d EEMC50 30-d EDEC50 [µg a.i./L ] Rate Rate Nominal Measured [Mean midge [Mean [mg a.i./L] [mg a.i./L] Start emerged] develoment (day 0) rate] 0 - 1.28 0.087 1.25 < LOQ 1.23 0.076 2.5 < LOQ 1.26 0.073 0.0114 > 0.010 5.0 < LOQ 1.25 0.075 10.0 9.0 1.22 0.075 20.0 16.0 0.0 - 40.0 33.0 0.0 - NOEC = 0.010 mg a.i./L LOQ: limit of quantification a EEMC50 : Endpoint Emergence rate / EDEC50: Endpoint Development rate Scenario B: Application to the Sediment

Table 7.4.3.5.1/01-2 Chronic toxicity of thiamethoxam to Chironomus riparius following application to the sediment

a a Concentration Emergence Development 30-d EEMC50 30-d EDEC50 [µg a.i./kg dry weight] Rate Rate Nominal Measured [Mean midge [Mean [mg a.i./kg [mg a.i./kg Start emerged] develoment sediment] sediment] (in sediment) rate] 0 - 1.29 0.070 12.5 < LOQ 1.31 0.066 25.0 < LOQ 1.06 0.068 0.11 > 0.10 50.0 < LOQ 1.13 0.071 100 < LOQ 1.08 0.071 200 < LOQ 0.0 - 400 160 0.0 - NOEC = 0.10 mg a.i./kg sediment LOQ: limit of quantification a EEMC50 : Endpoint Emergence rate / EDEC50: Endpoint Development rate

Observations: Emergence in the controls was greater than 80% and the mean development time for the larvae in exposure scenario A and B was not longer than 23 days. Conclusion: Exposure of Chironomus riparius to thiamethoxam, which was incorporated in the sediment simulating a run-off event, resulted in EC50 values of 0.11 mg a.i./kg for the emergence rate of larvae to adult midges and of > 0.10 mg a.i./kg sediment dry weight for the development rate of larvae. The NOEC was 0.10 mg a.i./kg sediment for both parameters.

If thiamethoxam is applied to the water phase the EC50 values for emergence and development rate in Chironomus riparius were 0.0114 and > 0.010 mg a.i./L water. The NOEC was 0.010 mg a.i./L.

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Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.3.5.1 Effects on sediment dwelling organism section No. / 02 91/414 Annex II Effects on sediment dwelling organisms Point addressed 8.2.7 / 02

1. Annex point(s) II A, 8.2.7 Effects on sediment dwelling organisms 2. Location in Dossier Section 6 3. Authors (Year) Grade, R. (1999)

Title Toxicity test of CGA 322704 (Metabolite of CGA 293343) on sediment-dwelling chironomus riparius (syn. Chironomus thummi) under static conditions.

Report No. / Date\ 982581 / 09.02.1999

Syngenta File N° 322704/021

Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Novartis Crop Protection, Inc., Ecotoxicology Laboratories, CH-4002, Basle, Switzerland 5. Dates of work 19th August 1998 through to 16th October, 1998 6. Test substance Company Code: CGA 322704, 7. Test method BBA Guideline Proposal, 1995, Effects of Plant Protection Products on Sediment-Dwelling Larvae of Chironomus riparius in a water-Sediment System. OECD Guideline for Testing of Chemicals, Proposal for toxicity Test with Chironomidae, May 1998. 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. OECD Principles of Good Laboratory, May 1981.

Test System: CGA 322704 (metabolite of thiamethoxam); . Test organism: Chironomus riparius larvae (syn. Chironomus thummi); Age: 2-3 days old 1st instar larvae. The scenario in which the compound is added to the sediment and which is intended to simulate a run-off event was selected as a most probable exposure scenario for metabolites. The test was performed in 1-L glass beakers containing approximately 1.6 cm sediment and a water column of 8 cm height at the beginning and of about 6 cm height at the end of the study. A constant temperature of 20 ± 2°C was maintained during the test with a photoperiod of 16 hours light and 8 hours dark. Exposure Scenario: CGA 322704 treated sand was mixed with aged artificial sediment at a range of concentrations (3.8, 7.5, 15, 30, 60 and 120 µg/kg sediment dry weight). The test concentrations were selected based on the results of a range finding study. The spiked sediment and water were added to the test vessels about 48 hours prior to the introduction of Chironomus larvae and the start of the test. Findings: Actual measured concentrations of CGA 322704 in the water phase at test day 0 after addition of the larvae were below the limit of detection in the two lowest test concentrations and 0.3, 0.8, 0.14 and 0.78 µg/L in the remaining groups. At the end of the study (test day 28) corresponding values were below the limit of detection for the two lowest doses and 0.3, 0.9, 4.4 and 6.5 µg/L for four higher dose levels. The sediment concentrations were measured in samples from the two highest administration rates i.e. 60 and 120 µg/kg sediment dry weight equivalent to 48 and 96 µg/kg sediment wet weight. At day 0, 7 and 28 the measured

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There were no indications on different sensitivities of sexes, therefore male and female results were pooled for statistical analysis. The biological assessment was based on impacts on full maturation of the la1vae to adult midges. Main parameters examined were the rate and time of emergence and the total number of emerging male and female midges.

Emergence in the control samples was greater than 80% and the mean development time for the la1vae was not longer than 23 days. Findings:

Table 7.4.3.5.1102-1 Chronic toxicity of CGA 322704 (metabolite of thiamethoxam) to Chironomus riparius following application to the sediment

Concentration Emergence Development 28-cl EEMCso a 28-d EoECso a CGA 322704 Rate Rate Nominal Measured [Mean midge [Mean of [mg/kg sediment] [mg/kg sediment] [µg/kg dry Start emerged] development weight (sum sediment rate per sediment] and water) vessel] (%nominal] 0 . 0.88 0.057 3.8 n.m. 0.90 0.060 7.5 n.m. 0.92 0.060 0.025 0.0529 15 n.m. 0.90 0.069

30 n.m. 0.12 0.037 60 62 0.00 . 120 82 0.00 . NOEC =0 .015 ma/ka sediment n m. = not measm·e.d a EEMCso : Endpoint Emergence rate I EoECso: Endpoint Development rate

Conclusion: Exposure of Chironomus riparius to CGA 322704 (metabolite of thiamethoxam) which was incorporated in the sediment simulating a nm-off event, resulted in EC50 values of 0.025 mg/kg for the emergence rate of laivae to adult midges and of 0.0529 mg/kg sediment diy weight for the development rate of laivae. The NOEC was 0. 01 5 mg CGA 322704/kg sediment for both parameters.

Results and discussion

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98/8 Doc IIIA 7.4.3.5.1 Effects on sediment dwelling organism section No. / 03 91/414 Annex II Effects on sediment dwelling organisms Point addressed 8.2.7 / 03

1. Annex point(s) II A, 8.2.7 Effects on sediment dwelling organisms 2. Location in Dossier Section 6 3. Authors (Year) Grade, R. (2000)

Title Toxicity Test of NOA 407475 (Metabolite of CGA 293343) on Sediment-Dwelling Chironomus riparius (syn. Chironomus thummi) under Static Conditions

Report No. / Date\ 982580 /July 12, 2000

Syngenta File N° 407475/ 14

Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Novartis Crop Protection, Inc., Ecotoxicology Laboratories, CH-4002, Basle, Switzerland 5. Dates of work 20th January, 1999 through to 28th May, 1999 6. Test substance Company Code: NOA 407475,

Test System: NOA 407475 (metabolite of thiamethoxam); Test organism: Chironomus riparius larvae (syn. Chironomus thummi); Age: 2-3 days old 1 instar larvae. The scenario in which the compound is added to the sediment and which is intended to simulate a run-off event was selected as a most probable exposure scenario for metabolites. The test was performed in 1-L glass beakers containing approximately 1.6 cm sediment and a water column of 8 cm height at the beginning and of about 6 cm height at the end of the study. A constant temperature of 20 ± 2°C was maintained during the test with a photoperiod of 16 hours light and 8 hours dark. Exposure Scenario: NOA 407475 treated sand was mixed with aged artificial sediment at a range of concentrations (32, 63, 125, 250, 500 and 1000 µg/kg sediment dry weight). The test concentrations were selected based on the results of a range finding study. The spiked sediment and water were added to the test vessels about 48 hours prior to the introduction of Chironomus larvae and the start of the test. Findings: Actual measured concentrations of NOA 407475 in the water phase were

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the measured test substance concentrations in the sediment (incl. interstitial water) were 91, 100, 100 and 100 µg/test vessel at a nominal concentration of 120 µg/test vessel. Total recovery from test system was 76, 83, 83 and 83 % of the nominal concentration at day –2, 0, 7 and 28.

There were no indications on different sensitivities of sexes at all test concentrations, therefore male and female results were pooled for statistical analysis. The biological assessment was based on impacts on full maturation of the larvae to adult midge. Main parameters examined were the rate and time of emergence and the total number of emerging male and female midges. Emergence in the control samples was greater than 80% and the mean development time for the larvae was not longer than 23 days. Table 7.4.3.5.1/03-1 Chronic toxicity of NOA 407475 (metabolite of thiamethoxam) to Chironomus riparius following application to the sediment

a a Concentration Emergence Development 28-d EEMC50 28-d EDEC50 NOA 407475 Rate Rate Nominal Measured [Mean midge [Mean of [mg/kg sediment] [mg/kg sediment] [µg/kg dry Start emerged] development weight (sum sediment rate per sediment] and water) vessel] [% nominal] 0 - 0.97 0.050 32 n.m. 0.97 0.051 63 n.m. 0.98 0.054 > 1.0 > 1.0 125 n.m. 0.95 0.052 250 n.m. 0.95 0.048 500 n.m 0.97 0.046 1000 83 0.85 0.058 NOEC = 1.0 mg/kg sediment n m. = not measured a EEMC50 : Endpoint Emergence rate / EDEC50: Endpoint Development rate

Conclusion: Exposure of Chironomus riparius to NOA 407475 (metabolite of thiamethoxam) which was incorporated in the sediment, resulted in EC50 values of >1.0 mg /kg for the emergence rate of larvae to adult midges and for the development rate of larvae. The NOEC was 1.0 mg /kg sediment for both parameters, the highest concentration tested.

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 27/12/2004 Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks COMMENTS FROM ... Date

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Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.4.3.5.1 Effects on sediment dwelling organism section No. / 04 91/414 Annex II Effects on sediment dwelling organisms Point addressed 8.2.7 / 04

1. Annex point(s) II A, 8.2.7 Effects on sediment dwelling organisms 2. Location in Dossier Section 6 3. Authors (Year) Grade, R. (2002)

Title Toxicity test of NOA 459602 (Metabolite of CGA 293343) on sediment-dwelling Chironomus riparius (syn. Chironomus thummi) under static conditions.

Report No. / Date\ 2012671 / 04.06.2002

Syngenta File N° 459602/0009

Source / Owner Unpublished / Syngenta Crop Protection AG

4. Testing facility Syngenta Crop Protection AG. CH-4002, Basel, Switzerland 5. Dates of work 14th February 2002 through to 11th April 2002 6. Test substance Company Code: NOA 459602,

7. Test method Effects of Plant Protection Products on the Development of Sediment Dwelling Larvae of Chironomus riparius in a Water-Sediment System (BBA Guideline Proposal 1995) OECD Guideline for Testing of Chemicals, Proposal for a new Guideline 219, Sediment- water Chironomid toxicity test using spiked water, February 2001. 8. Deviations None

9. GLP With the exception of the range finding test, characterisation of sediment and development of analytical method, this study was performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted February 2nd, 2000 (RS 813.016.5).

This Ordinance is based on the OECD Principles of Good Laboratory Practice, as revised in 1997 and adopted November 26th, 1997 by decision of the OECD Council [C (97) 186 (Final)].

Test System: NOA 459602 (metabolite of thiamethoxam); Test organism: Chironomus riparius larvae (syn. Chironomus thummi); Age: 2-3 days old 1st instar larvae. The scenario in which the compound is added to the water and which is intended to simulate a spray-drift event was selected as a most probable exposure scenario for this metabolite. The test was performed in 1-L glass beakers containing approximately 1.6 cm sediment and a water column of approximtely 8 cm height at the beginning. Three replicates were set up to study emergence, whilst an additional replicate was included to determine larval survival and growth by day 10. A constant temperature of 21.0 – 21.7°C was maintained during the test with a photoperiod of 16 hours light and 8 hours dark and a light intensity of approximately 1000 lux. The conductivity was 790 and 785 μS/cm and the water hardness corresponded to 282 and 280 mg CaCO3/L (two bottles). Exposure Scenario: NOA 459602 stock solutions were added to the water column and the upper water layer was gently mixed without disrurbing the aged artificial sediment to give a range of concentrations (3.13, 6.25, 12.5, 50 and 100 mg/L). The test concentrations were selected based on the results of a range finding study. The water column was treated one day after the introduction of Chironomus larvae (20 larvae per test vessel).

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Findings: Actual measured concentrations of NOA 459602 were 3.3, 7.1, 13.7, 53.3 and 106.9 mg/L, corresponding to 105.4, 113.6, 109.6, 106.6 and 106.9 % of nominal concentrations at the beginning of the test. At the end of the study (day 24) measured concentrations of NOA 459602 in the water phase were 2.7, 4.5, 10.9, 40.2 and 82.0 mg/L, corresponding to 86.3, 72.0, 87.2, 80.4 and 82.0 % of nominal concentrations at the beginning of the test. NOA 459602 concentrations in the sediment and the interstitial water was measured only for the highest test concentration and the lower one and the control. The amount in the sediment increased from 0.84 to 8.6 and 7.8 % of total nominal concentration of 50 mg/L at day 0, 7 and 24, respectively. For nominal concentration of 100 mg/L the amount in the sediment increased from 1.0 to 7.7 and 7.9 of total on these respective sampling days.

Oxygen content ranged from 5.1 to 6.8 mg/L just after application of the test item to test vessels. At test termination the oxygen content was within a range of 8.0 to 9.8 mg/L. The pH increased from 7.1 to 7.4 just after application to values ranging from 8.6 to 9.2 at test termination.

There were no indications of different sensitivities of sexes at all test concentrations, therefore male and female results were pooled for statistical analysis. The biological assessment was based on impacts on full maturation of the larvae to adult midge. Main parameters examined were the rate and time of emergence and the total number of emerging male and female midges. Growth measurements of the larvae via the estimation of the weight was carried out. After 10 days of exposure 17, 18, 20, 16, 20 and 19 larvae were found at blank and at test concentrations of 3.13, 6.25, 12.5, 50 and 100 mg/L. The corresponding average dry weight of the test groups were 1.43, 1.49, 1.38, 1.48, 1.25 and 1.39 mg/larvae.

Emergence in the control samples was greater than 80% and the mean development time for the larvae was not longer than 23 days. Table 7.4.3.5.1/04-1 Chronic toxicity of NOA 459602 (metabolite of thiamethoxam) to Chironomus riparius following application to the water phase

a a Concentration Emergence Development 24-d EEMC50 24-d EDEC50 NOA 459602 Rate Rate Nominal in Measured in [Mean midge [Mean of [mg/L] [mg/L] water phase water phase at emerged/ development [mg/L] a start introduced rate per [% nominal] larvae] vessel] 0 - 0.98 0.072 3.13 105.4 0.90 0.074 6.25 113.6 0.93 0.072 56.0 n.d. 12.5 109.6 0.88 0.068 50 106.6 0.90 0.075 100 106.9 0.0 - NOEC = 50 mg/L n.d. = not determined a EEMC50 : Endpoint Emergence rate / EDEC50: Endpoint Development rate

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Conclusion: Exposure of Chironomus riparius to NOA 459602 (metabolite of thiamethoxam) which was inco1porated in the water phase, resulted in EC50 values of 56.0 mg/L for the emergence rate of laivae to adult midges. No EC50 value for the development rate of laivae could be calculated. The NOEC for emergence rate and development rate were 50 mg /L for both pai·ameters.

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Section A 7.4.3.5.1 Effects on sediment dwelling organisms Annex Point IIIA XIII.3.4

Official 1 REFERENCE use only 1.1 Reference Smyth, D.V., Brown, R.J., Maynard, S.J., (2004); CGA 322704 (Thiamethoxam metabolite): Toxicity to the sediment dweller Chironomus 1·iparius using spiked water. Brixham Environmental Laborat01y, Brixham, Devon, England, Repo1t No.: BL7987/B (Syngenta Project No. 2033605), 2 December 2004 (unpublished). 1.2 Data protection Yes. 1.2.1 Data owner Syngenta Crop Protection 1.2.2 Companies with letter of access .. 1.2.3 Criteria for data protection

2 GUIDELINES AND QUALITY ASSURANCE 2.1 Guideline study Proposal for a new OECD Guideline 219: "Sediment-Water Chironomid Toxicity Test Using Spiked Water" (Februaiy 2001). 2.2 GLP Yes (ce1tified laboratory). 2.3 Deviations None.

3 METHOD 3.1 Test material CGA 322704 (metabolite ofCGA 293343) 3 .1.1 Lot/Batch number ..

3.1.2 Purity - 3 .1.3 Fwther relevant Not applicable x prope1ties 3.2 Preparation of TS A primary stock solution was prepared by direct addition to ASTM solution for poorly 'hard' water (prepared at half strength). Secondaty stock solutions were soluble or volatile prepared by using either aliquots of the primary stock or a higher test substances concentration secondaty stock solutions to dilution water 3.3 Reference Not used in this study. x substance 3.4 Testing procedure 3.4.1 Dilution water and See Table A 7.4.3.5.1-1 sediment 3.4.2 Test organisms See Table A 7.4.3.5.1-2 3.4.3 Test system See Table A 7.4.3.5.1-3 3.4.4 Test conditions See Table A 7.4.3.5.1-4 3.4.5 Dmation of the test 28 days. 3.4.6 Test parameter Emerged midges, development rate. 3.4.7 Examination I Sex, time and number of emerged adults were observed daily dming

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Section A 7.4.3.5.1 Effects on sediment dwelling organisms Annex Point IIIA XIII.3.4 Sampling period of emergence. 3.4.8 Monitoring of TS Yes. 0, 7, 28 days. concentration 3.4.9 Statistics Nonnality analysis (Shapiro and Wilks, 1965), homogeneity of variance (Snedecor and Cochran, 1980), parametric analysis (Hayslett and Mmphy, 1976), Mann Whitney U-test (Zar, 1999), probit analysis (Stephan, 1977).

4 RESULTS 4.1 Range finding test Not mentioned. 4.2 Results test substance 4.2.1 Initial 0.073, 0.22, 0.67, 2.0, 6.0 and 18 µg/L (nominal initial). concentrations of test substance See Table A 7.4.3.5.1-5. 4.2.2 Effect data See Table A 7.4.3.5.1-6,7,8. 4.3 Results of controls See Table A 7.4.3.5.1-5,6,7,8 4.4 Test with Not perfo1med in this study. x reference substance

5 APPLICANT'S SUMMARY AND CONCLUSION 5.1 Materials and The influence of CGA 322704 (thiamethoxam metabolite) on methods development and emergence of larvae of Chironomus ripmius in a water-sediment system was investigated according to the new OECD Guideline 219: "Sediment-Water Chironomid Toxicity Test Using Spiked Water" (Febmaiy 2001). Larvae of Chironomus riparius (1st instars < 48 hours old, 4 beakers per test concentration and control with 20 animals each) were exposed for 28 days in a static test system to concentrations of0.073, 0.22, 0.67, 2.0, 6.0 and 18 µg/L (nominal initial) in a water-sediment system (spiked water). 5.2 Results and The validity criteria of the study were met, see table A7.4.3.5.1J9 discussion The analytical results for the test concentrations of 0.22, 0.67, 2.0, 6.0 and 18 µg/L were 0.21, 0.69, 2.1, 4.9, and 15 µg/L, coITesponding to 95, 103, 105, 82 and 83% of nominal concentrations on day 0.

See table A 7.4.3.5.1 -00.

The results of the study are based on the nominal exposure concentrations.

No emergence occUITed at test concentrations of 6.0 and 18 ~1g IL. Total numbers of emerged tnidges were significantly lower in the 2.0 µg/L treatment.

The EC50 based on the number of emerged midges after 28 days was calculated to be 1.2 µg/L (95% confidence intervals of 0.96 to 1.5 µg/L).

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Section A 7.4.3.5.1 Effects on sediment dwelling organisms Annex Point IIIA XIII.3.4

The nominal test concentrations of0.67 µg/L and below were assessed as having no effect on chi.ronomid emergence (P=0.05) based on the criteria time to first emergence, mean emergence time, number emerged after 28 days and the sex ratio of adults. 5.2.1 NOEC Time to first emergence - 2.0 µg/L Mean emergence time - 2.0 ~1g/L Number emerged after 28 days - 0.67 µg/L Sex ratio of adults - 0.67 ~1g/L 5.3 Conclusion The overall no-observed effect concentration (NOEC) for the study was 0.67 µg/L 5.3.1 Reliability 1 5.3.2 Deficiencies No

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Section A 7.4.3.5.1 Effects on sediment dwelling organisms Annex Point IIIA XIII.3.4 Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted

EVALUATION BY RAPPORTEUR MEMBER STATE Date l l/4/2006 Materials and Methods

Results and discussion Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... (specify) Date Giw date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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Table A 7.4.3.5.1-1: Dilution water and sediment Criteria Details Water ASTM ‘hard’water (half strength) Sediment artificial (75% quartz sand, 5% peat, 20% kaolin, pH = 6.0 ± 0.5) Measured organic matter content of sediment 4.3% Organic content of sediment 2.5%

Hardness 77 mg/L CaCO3 pH 8.08 Ca / Mg ratio Not specified. Na / K ratio Not specified. Oxygen content 9.0 mg/L. Conductance 304 µS/cm. TOC 2.452 mg/L Holding water different from dilution water Not applicable.

Table A 7.4.3.5.1-2: Test organisms Criteria Details Strain / Clone Chironomus riparius. Source Laboratory culture Age Larvae <48 hour post-hatch Breeding method Not stated in the report Kind of food Fish food extract (Aquarian® Tropical Fish Flakes). Amount of food Not stated in the report Feeding frequency Not stated in the report Pretreatment Not stated in the report Feeding of animals during test Yes. Once daily days -1 to 5 each vessel recieved 5.0 mg of food, for days 6 to 9 each received 20 mg of food, for days 10 and 11 each received 10 mg of food, for the remainder of the study each revieved 5 mg per day.

Table A 7.4.3.5.1-3 Test system Criteria Details Test type Static Volume of test vessels 0.4 L glass beakers fitted with lids (inverted plastic beakers with meshed air vents). Each vessel contained a sediment layer (17 mm, equivalent to 120 g dry weight) and overlying water (292 mL of dilution water, depth of 68 mm)in a 1:4 ratio Number of animals/vessel 20 (except replicate D, 0.073 µg/Lwhich recieved 30 and replicate D, 0.022 µg/Lwhich received 10)

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Number of vessels/ concentration 6 per treatment (four biological replicates and two analytical replicates) (dilution water control), 0.073, 0.22, 0.67, 2.0, 6.0 and 18 µg/L

Table A 7.4.3.5.1-4: Test conditions Criteria Details Test temperature 19.7 to 20.5°C

Dissolved oxygen 8.0 to 8.9 mg O2/L pH 7.56 to 8.08 Adjustment of pH Not necessary Aeration of dilution water Yes, gentle Quality/Intensity of irradiation Not stated in the report Photoperiod 16 h photoperiod daily with a 20 minute transition period

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Table A 7.4.3.5.1-5: Analysis of CGA 322704 (thiamethoxam metabolite) in overlying water Initial nominal Measured CGA 322704 concentrations (µg/L) concentration (µg/L) Day 0 Day 7 Day 28

Control <0.046 <0.074 <0.022 0.073 <0.047 <0074 <0.023 0.22 0.21 <0.072 <0.023 0.67 0.69 0.41 <0.020 2.0 2.1 1.1 <0.022 6.0 4.9 <3.0 <1.2 18 15a 6.7a 5.8a (15, 15, 15) (6.2, 7.0, 6.8) (6.2, 5.4, 5.9) a mean of triplicate analysis

Table A 7.4.3.5.1-6: Influence on the emergence and development of Chironomus riparius after 28 days (based on nominal initial concentrations of CGA322704) Initial nominal Emergence of inserted larvae Development concentration (pooled sex) (µg/L) Time to first Mean emergence Mean total (%) emergence (d) time (d) Mean rate (1/d) Control 14 15.6 89 0.06405 0.073 14 15.2 93 0.06597 0.22 14.3 16.0 93 0.06256 0.67 14 15.2 95 0.06568 2.0 14 14.3 28 0.06988 6.0 - - 0 - 18 - - 0 - - no emergence

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Table A 7.4.3.5.1-7: Sex ratio of emerged adults Initial nominal % males concentration (µg/L) Replicate A Replicate B Replicate C Replicate D Mean Control 39 50 56 63 52 0.073 61 68 63 NR 64 0.22 39 26 68 NR 45 0.67 45 50 44 50 47 2.0 75* 71* 60* 83* 72* 6.0 - - - - - 18 - - - - - NR – replicate value not reported and excluded from statistical determination, error in number of midges added * - significantly different to the control (P=0.05) - no emergence

Table A 7.4.3.5.1-8: Influence on the emergence and development after 28 days (based on nominal initial concentrations)

Parameter EC50 (µg/L) 95% confidence No-observed effect interval concentration (P=0.05) (µg/L) Time to first emergence na na 2.0 Mean emergence time na na 2.0 Number emerged after 1.2 0.96 – 1.5 0.67 28 days Sex ratio of adults na na 0.67

Table A 7.4.3.5.1-9: Validity criteria for invertebrate reproduction test according to OECD Guideline 219 Criterion fulfilled Not fullfilled Mortality in the controls < 30% at test termination Yes Emergence in controls should be 12 to 23 days Yes Dissolved oxygen concentration should be at least 60% ASV and pH 6 Yes to 9 in all test vessels Water temperature should not differ by more than ± 1°C between test Yes vessels

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98/8 Doc IIIA 7.4.3.5.2 Aquatic plant toxicity section No. / 01 91/414 Annex II Aquatic plants Point addressed 8.2.8 / 01

1. Annex point(s) II A, 8.2.8 Aquatic Plants 2. Location in Dossier Section 6 3. Authors / Year Grade, R. (1998c)

Title 6 ACUTE TOXICITY TEST OF CGA-293343 TECH. TO THE DUCKWEED LEMNA GIBBA G3 UNDER SEMI-STATIC CONDITIONS Report No. / Date\ 972561 / 16.06.1998 Novartis File N° Novartis Study # 293343-595 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility Novartis Crop Protection AG, Ecotoxicology Department, CH-4002 Basle Switzerland 5. Dates of work October 17, 1997 through November 24, 1997 6. Test substance ISO common name thiamethoxam. Company Code: CGA 293343 tech.,

7. Test method ASTM Guideline E1415-91, Lemna gibba. US-EPA FIFRA Guideline Number 122-2 and 123-2 OECD Guideline (Draft proposal, July 1996) OPPTS Guideline, 850.4400, Lemna gibba. EPA ecological test guidelines, Public Draft, April 1996. 8. Deviations None 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by: 1) Good Laboratory Practice in Switzerland, Procedures and Principles, March 1986. 1) OECD Principles of Good Laboratory, May 1981. 1) U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989 1) Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984.

Test System: Thiamethoxam technical; Fronds of Lemna gibba (strain G3) were exposed for 7 days to CGA 299343 in a static test system with renewal of the test solution after 3 days. Six concentrations ranging from nominal 3.13 to 100 mg/L were employed with three replicates per treatment level and the controls. The test was initiated by placing asceptically five plants with 2-5 fronds into each test vessel. At intervals of 3 days and at test termination fronds were counted and biological observations were made. The results are based on mean measured concentrations and are reported as the 7-day NOErC and ErC50 values (the concentrations of test substance that reduced frond number and growth rate by 50% as compared to the control).

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Findings:

Acute toxicity of thiamethoxam to aquatic higher plants

Frond production, relative to control, based upon frond number (growth rate) and frond drv weiaht (biomass\ on Dav 7 Concentration Mean Frond Mean Frond Reduction Exposure EC-so Number dry weight in growth period (95 % conf. Ima a.i./Ll (biomass) rate 8 interval) Nominal Mean (mean indiv. (mg dry (%) (days) (mg a.i./L) Measure counts) weight) d Control . 181 29.6 0.00 3.13 2.9 169 25.1 4.51 6.25 5.7 184 29.3 0.04 7-day > 90.2 12.5 11.4 172 25.9 4.14 25 22.0 177 26.0 2.45 50 43.9 185 31.5 0.91 100 90.2 177 27.1 2.65 NOEC: 90.2 mg a.i./L a promot10n oflemna growth with regard to control Conclusion: The 7-day ErCso value for frond density was dete1m ined to be > 90.2 mg a.i./L, based on mean measured concentrations, and > 100 mg a.i./L based on nominal values. The EC50, expressed as the reduction in frond biomass (diy weight), was also> 90.2 mg a.i./L. The NOEC for effects on frond density and biomass was found to be 90.2 mg a.i./L, the highest concentration tested.

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted EVALUATION BY RAPPORTEUR MEMBER STATE Date 28/ 12/2004 Materials and M ethods Results and discussion Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and M ethods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc lllA 7.5.1.1 Inhibition of microbiological activity section No. 01 91 /414 Annex II Effects on soil non-target micro-organisms Point addressed 8.5 / 01

1. Annex point(s) II A, 8.5 Effects on soil non-target micro-organisms 2. Location in Dossier Section 6, 3. Authors (year) Bader, U. (1998): Title The effect of CGA 293343 tech. on soil respiration and nitrification

Report No., Date 972515, 24April, 1998 Novartis File N° (Desire) 293343/532 Owner Novartis Crop Protection AG 4. Testing facility Novartis Crop Protection AG Basel Switzerland 5. Dates of work Study Initiation: 2 July 1997 Experimental Start: 14 January 1998 Experimental Termination: 27 Febmary 1998 Study Completion: 24 April, 1998 6. Test substance ISO common name thiamethoxam.

7. Test method BBA Guidelines, Part VI, 1-1 (2. edition) March 1990. OECD Guidelines for Testing of Chemicals (Draft June 1996), Soil Microorganisms, Carbon and Nitrogen Mineralisation.

SETAC-Europe, Procedm·es for Assessing the Environmental Fate and Ecotoxicity of Pesticides (March 1995). Deviations: Measuring of anunonitun, nitrite and nitrate concentrations in soil was pe1fon11ed after amending the soil with powered Lucerne, the respiration rate was measured in soil not amended with Lucerne meal. 8. Deviations none 9. GLP Yes , Good Laborato1y Pra.ctice (GLP) in Switzerland, Procedm·es and principles, March 1986, issued by the Federal department of the Interior and the Intercantonal Office for the Control of Medicaments, Switzerland. These are based on the OECD Principles of Good Laborato1y Pra.ctice, adopted 12 May, 1981 by the decision of the OECD Council [C (81) 30 (Final)] concerning Mutual Acceptance of Data in the Assessment of Chemicals, the Recommendation of the Council of26 July, 1983 concerning the Mutual Recognition of Compliance with Good Laboratory Practice [C (83) 95 (Final)], and the Council Decision Recommendation of2 October, 1989 concerning Compliance with Principles of Good Laborato1y Pra.ctice Council [C (89) 87 (Final)]. With the exception of the determination of soil properties by Agrolab AG, CH-6030 Ebikon.

Test System: Thiamethoxam tech. The possible effects of thiamethoxam on soil micro-organisms were etemune y measunng the rate of sho1t-te1m respiration and nitrogen conversions (nitrogen mineralisation and nitiification) in soil ti·eated with n01ninal test concenti·ations of 0.27 and 2.67 mg thiamethoxamlkg illy soil (10 times the lower test concenti·ation). The Inicrobial biomass of the loamy sand, (0.99 % organic carbon and 71.86 % sand, pH 7.5), was measured p1ior to applying the test substance by substi·ate-induced respiration. The biomass was calculated from the constant initial respiration rate. It was dete1mined to be 67 mg C per 100 g illy soil and confnm adequate Inicrobial activity of the soil. Based on these results 1000

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mg glucose per 100 g soil dry weight was added to the soil samples and the initial short-term respiration rates were measured over a period of approximately 24 hours. Soil respiration was measured over a period of 28 days. Nitrogen conversion was determined by measuring the content of NH4-N and NO3-N (NO3 and NO2-N) in soil amended with lucerne meal as a nitrogen source. Ammonium-N nitrite-N and nitrate-N were determined after 0-3 hours, 14 days and 28 days after application. The test method was calibrated with the reference substance Dinoseb.

Findings: Rates of short-term respiration measured during the constant initial phase of respiration (% deviation of treated samples from the control) Samples Control 0.27 mg CGA 2.7 mg CGA 293343/kg Dinoseb 293343/kg dry soil dry soil

[mg CO2/h] [mg CO2/h] % [mg CO2/h] % [mg CO2/h] % 0-3 hours 1.858 1.882 1.3 1.845 -0.7 1.669 -10.2 day 14 2.067 2.000 -3.2 1.971 -4.6 1.396 -32.5 day 28 1.633 1.587 -2.8 1.524 -6.7 0.960 -41.2

After 0, 14 and 28 days of exposure no significant test substance induced effects on soil short term respiration rates were observed.

NO3-N, NH4-N and total-N concentrations in thiamethoxam- and dinoseb treated soil: Average concentrations in mg/100g soil dry weight at the lower and higher test item concentrations

Samples Incubation 0-3 hours 14 Days 28 Days

NO3-N NH4-N Total-N NO3-N NH4-N Total-N NO3-N NH4-N Total-N [mg/100g soil dry weight] Control 4.70 0.93 5.7 6.50 1.30 7.8 8.15 0.55 8.7 0.27 mg 4.57 0.90 5.6 6.33 0.93 7.3 8.23 0.40 8.6 CGA 293343/kg a 2.7 mg 4.47 0.97 5.5 6.07 0.87 6.9 7.97 0.43 8.4 CGA 293343/kg a Dinoseb 4.040 0.87 5.4 8.13 0.50 8.6 10.03 0.37 10.4 a kg dry weight of soil

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N0 3-N, NH4-N and total-N concentrations in thiamethoxam- and dinoseb treated soil: % deviation of treated samples from the control at the lower and higher test item concentrations

Samples Incubation Day

0-3 hours 14 Days 28 Days [Total N, % deviation from control) Control --- 0.27 mg CGA 293343/kg a -1.8 -6.4 -1.1

2.7 mg CGA 293343/kg a -3.5 -1 l.5 -3.4 Dinoseb -5.3 10.3 19.5 a kg diy weight of soil

In the loamy sand total-N concentrations over the whole incubation pe1iod, i.e. after 0, 14 and 28 days exposure to thiamethoxam, deviated by less than 12 % from the control at 0.27 mg thiamethoxam and 2.7 mg thiamethoxamlkg illy weight soil.

Conclusion: Thiamethoxam had no significant effects on soil sho1t-te1m respiration rates or on nitrogen conversion up to a dose rate of 2.67 mg/kg illy soil. At test te1mination, 28 days after the treatment with thiamethoxam, sho1t-te1m respiration and nitrogen conversion in soil differed by less than 12% from the control at all treatment rates. Thiamethoxam at the concentrations tested, therefore had, no adverse effects on organic matter turnover and hence on soil fe1t ility according to the classification scheme given by Malkomes 199028.

Results and discussion Conclusion Reliability I Acceptability -

28Malkomes, H.P. 1990/03 : Richtlinien fur die amtliche P1iifung von Pflanzenschutzmitteln Teil VIl- 1. 2. Auflage, Auswirkungen auf die Aktivitat der Bodenmikroflora.

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Remarks COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.5.1.1 / Inhibition of microbiological activity section No. 02 91/414 Annex II Effects on soil non-target micro-organisms Point addressed 8.5 / 01

1. Annex point(s) II A, 8.5 Effects on soil non-target micro-organisms 2. Location in Dossier Section 6, 3. Authors (Year) Bader, U. (1999): Title The effect of CGA 322704 + CGA 355190 (two Metabolites of CGA 293343) on soil respiration and nitrification

Report No., Date 992668 / 11.11.1999

Syngenta File N° 322704/0023

Source / Owner Unpublished / Syngenta Crop Protection AG 4. Testing facility Novartis Crop Protection AG, Basel, Switzerland 5. Dates of work 19th May 1999 through to 16th August 1999 6. Test substance Company Code: CGA 322704., Company Code: CGA 355190., 7. Test method OECD Guidelines for Testing of Chemicals (revised draft January 1999), Proposal for a new Guideline 216, Soil Micro-organisms, Nitrogen Transformation test and Proposal for a new Guideline 217, Soil Micro-organisms, Carbon Transformation Test. 8. Deviations None 9. GLP Good Laboratory Practice (GLP) in Switzerland, Procedures and principles, March 1986, issued by the Federal department of the Interior and the Intercantonal Office for the Control of Medicaments, Switzerland. These are based on the OECD Principles of Good Laboratory Practice , adopted 12 May, 1981 by the decision of the OECD Council [C (81) 30 (Final)] concerning Mutual Acceptance of Data in the Assessment of Chemicals, the Recommendation of the Council of 26 July, 1983 concerning the Mutual Recognition of Compliance with Good Laboratory Practice [C (83) 95 (Final)], and the Council Decision- Recommendation of 2 October, 1989 concerning Compliance with Principles of Good Laboratory Practice Council [C (89) 87 (Final)]. With the exception of the determination of soil properties by Agrolab AG, CH-6030 Ebikon.

Test System: The possible effects of two metabolites of thiamethoxam, CGA 322704 and CGA 355190, on soil micro-organisms were determined by measuring the rate of short-term respiration and nitrogen conversions (nitrogen mineralisation and nitrification) in soil treated with nominal test concentrations of 0.1 and 0.5 mg of both CGA 322704 and of CGA 355190 /kg dry soil. The microbial biomass of the loamy sand (1.78-1.86% OC, 56.4-64.2% sand) was measured prior to applying the test substance by substrate-induced respiration. The biomass was calculated from the constant initial respiration rate. It was determined to be 27.2 mg C per 100 g dry soil used for the respiration test and 23.5 mg in the soil used for the nitrogen transformation test. This confirms adequate microbial activity of the soil. Based on these results 400 mg glucose per 100 g soil dry weight were added to the soil samples and the initial short-term respiration rates were measured over a period of approximately 24 hours. Soil respiration samples were taken after incubation periods of 1-3 hours, 7, 14 and 28 days at 20 ± 1° C in the dark. Nitrogen conversion was determined by measuring the content of ammonium nitrogen (NH4-N) and nitrate and nitrite nitrogen (NO3- and NO2-N) in soil amended with lucerne meal as a

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nitrogen source. Ammonium-N, nitrite-N and nitrate-N were determined after 1-3 hours, 7 days, 14 days and 28 days after application. The test method was calibrated with the reference substance Dinoseb.

Findings: Rates of short-term respiration measured during the constant initial phase of respiration (% deviation of CGA 322704 + CGA 355190 treated samples from the control) Samples Control 0.1 mg/kg dry soil 0.5 mg/kg dry soil Dinoseb

[mg CO2/h] [mg CO2/h] % [mg CO2/h] % [mg CO2/h] % 1-3 hours 11.57 10.86 -6.1 9.63 -16.8 10.21 -11.8 day 7 11.49 11.04 -3.9 10.35 -9.9 8.33 -27.5 day 14 9.35 9.82 5.0 9.37 0.2 6.29 -32.7 day 28 10.55 9.86 -6.5 9.47 -10.2 5.72 -45.8

After 0, 7, 14 and 28 days of exposure no significant test item induced effects on soil short term respiration rates were observed for both application rates.

NO3-N, NH4-N, NO2-N and total-N concentrations in CGA 322704 + CGA 355190 - and in dinoseb treated soil: average concentrations in mg/kg soil dry weight at the lower and higher test item concentrations Samples Control 0.1 mg/kg dry soil 0.5 mg/kg dry soil Dinoseb

1- 3 hours NO3-N 17.7 17.8 17.8 17.8

NH4-N 7.5 10.0 10.0 9.0

NO2-N 0.3 0.2 0.5 0.8 Total-N 25.5 28.0 28.3 27.6

7 days NO3-N 27.1 19.4 16.4 37.3

NH4-N 4.8 4.7 4.7 5.0

NO2-N 0.2 0.2 0.2 0.7 Total-N 32.1 24.3 21.3 43.0

14 days NO3-N 40.2 35.5 31.5 62.5

NH4-N 3.0 3.7 3.3 3.3

NO2-N 0.2 0.2 0.2 0.2 Total-N 43.4 39.4 35.0 66.0

28 days NO3-N 55.6 51.1 50.5 80.1

NH4-N 1.5 1.7 2.0 1.3

NO2-N 0.2 0.2 0.2 0.2 Total-N 57.3 53.0 52.7 81.6 - - + Total N = sum of NO3 , NO2 and NH4

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Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc lllA 7.5.1.2 Acute toxicity to earthworm and other non-target macro-organism section No. 01 91 /414 Annex II Effects on earthworms: Acute toxicity Point addressed 8.4.1 /01

1. Annex point(s) II A, 8.4.1 Effects on earthworms: Acute toxicity 2. Location in Dossier Section 6, 3. Authors (year) Candolfi, M.P. (1995) Title CGA 293343 tech: 14-day acute toxicity test with the earthworm (Eiseniaf oetida). Report No., Date 95-065-1008, 28 November, 1995 Novartis File N° (Desire) 293343/23 Owner Novartis Crop Protection AG 4. Testing facility Springbom Laboratories AG Hom Switzerland 5. Dates of work Study Initiation: 4 September 1995 Experimental Start: 25 September 1995 Experimental Tel'lllination: IO October 1995 Study Completion: 28 November, 1995 6. Test substance ISO common name thiamethoxam.

7. Test method OECD-Guideline No. 207, 1984, Earthwonn, Acute Toxicity Test 8. Deviations none 9. GLP Yes , Performed in compliance with all the pertinent OECD and Swiss Good Laboratory Practice Standards (OECD, 1992; Eidg. Dept. des Innem, 1988) with the following exception: routine soil contaminant screening analysis for pesticides, PCBs and metals were conducted using standard U.S. EPA procedures by Lancaster Laboratories, Lancaster, PA.

Test System: Thiamethoxam tech.; . Test organism: earthwo1m (Eiseniafoetidafoetida); Age: mature a u ts wit c um > 2 months old); weight range of eruthwo1ms 300 to 600 mg; Source: Gier-Angel, Neuenhof, Switzerland. Eruthwo1ms were dosed by unifo1mly mixing the test substance into an a1tificial soil substrate which was then dispensed into each test vessel. The moisture content of the soil was 34 to 35% and the pH was 5.5 to 5.6. The study was a limit test with one test substance concentration of 1000 mg a.i./kg soil diy weight and one deionized water control (artificial soil substrate). Expe1imental design: 40 eaithwo1ms with 4 replicates per treatment (each replicate with 10 eaithwo1ms). Test duration: 14 days. Assessments on m01tality and abno1mal behaviour were made at 7 and 14 days after treatment. The wo1ms were weighed at the beginning and end of the test.

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Findings: Acute toxicity of thiamethoxam to earthworms

Mortality Mean Mean live Mean Concentratio Burrowing weight change in Exposur LC so n [%) time [mg) live e period (95 % cont. lminutesl weiaht 1%1 interval) [mg a.i./kg 7d 14 d Od 7d Od 14 d 0 - 14 d [days] [mg a.i./kg soil] soil] Control 0.0 0.0 4.5 4.0 349. 361. 3.4 14 > 1000 (none) 3 2 1000 5.0 7.5 4.3 8.3 342. 278. -18.6 14-d NOEC: <1000 mg 3 5 a.i./kg soil

Observations: A significant higher bunowing time of eruthwo1ms in the test item group (8.3 minutes) was noted compru·ed to eruthwo1ms from the control group (4 minutes). Ea1thwo1ms exposed to the control soil remained stable in weight over the 14 day exposure period, while, erut hwo1ms exposed to thiamethoxam lost weight (- 18.6%).

Conclusion: The 14-day LC50 for erut hwo1ms exposed to thiamethoxam was dete1mined to be > 1000 mg a.i./kg diy weight soil and the overall NOEC was dete1mined to be < 1000 mg a.i./kg diy weight soil (the only concentration tested).

Results and discussion

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Conclusion

Reliability Acceptability Remarks COMMENTS FROM ... Date Materials and Methods Results and discussion Conclusion Reliability Acceptability Remarks

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98/8 Doc IIIA 7.5.1.2 / Acute toxicity to earthworm and other non-target macro-organism section No. 02 91/414 Annex II Effects on earthworms: Acute toxicity Point addressed 8.4.1 / 02

1. Annex point(s) II A, 8.4.1 Effects on earthworms: Acute toxicity 2. Location in Dossier Section 6, 3. Authors (Year) Bryan, R.L. (1999a) Title NOA 407475: An acute toxicity study with the Earthworm in an artificial soil substrate. Report No. / Date 108-417 / 09.12.1999

Syngenta File N° 407475/0013 Source / Owner Unpublished / Syngenta Crop Protection AG 4. Testing facility Wildlife International, Ltd., 8598 Commerce Drive, Easton, Maryland 21601 5. Dates of work 14th October 1999 through to 28th October 1999 6. Test substance Company Code: NOA 407475,

7. Test method OECD-Guideline No. 207, 1984, Earthworm, Acute Toxicity Test 8. Deviations Verification of the test concentrations, stability and homogeneity in the soil were not determined. 9. GLP Performed in compliance with all the pertinent Good Laboratory Practice Standards U.S. Environmental Protection Agency 40 CFR Parts 160 and 792, 17 August 1989, OECD and Japan MAFF) with the following exception : The stability of the test substance under storage conditions at the site was not conducted in accordance with GLP.

Test System: The acute effects of NOA 407475, metabolite of thiamethoxam; on earthworms was tested.. Earthworms (Eisenia foetida foetida); of mature age with an average individual body weight 310 to 360 mg; Source: Willingham Worm Farm, Butler Georgia. Earthworms were dosed by uniformly mixing the test substance into an artificial soil substrate which was then dispensed into each test vessel. The moisture content of the soil was 33% approximately and the pH at preparation on day 0 was between 7.8 to 8.1. The worms were exposed to five test concentrations of 62.5, 125, 250, 500 and 1000 mg NOA 407475/kg of dry soil. Four replicate test chambers were maintained in each treatment and control group, with 10 worms in each chamber. Assessments on mortality and abnormal behaviour were made at 7 and 14 days after treatment. The worms were weighed at the beginning and end of the test and were not fed during testing. A reference toxicity test was conducted with chloroacetamide.

Findings: Acute toxicity of NOA 407475 (metabolite of thiamethoxam) to earthworms

Mortality Mean live Mean Concentration weight change in Exposure LC50 of NOA 407475 [%] [mg] live period (95 % conf. weight [%] interval) [mg /kg soil] 7 d 14 d 0 d 14 d 0 - 14 d [days] [mg /kg soil] Control 0.0 0.0 360 210 -41.7 62.5 0.0 0.0 320 220 -31.3 125 0.0 0.0 310 200 -35.5 14 >1000 (none) 250 0.0 0.0 310 190 -38.7 500 0.0 0.0 320 180 -43.8

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1000 0.0 0.0 310 180 -41.9 14-d NOEC: 125 mg /kg soil

Observations: There was no apparent aversion to the soil in any of the treatment groups. All worms had burrowed into the soil within 75 minutes. Bodyweight change during the test was not statistically significant (p>0.05) at any concentration tested in comparison to the control group. There was some thinning and reduced reaction to external stimuli observed in the 250, 500 and 1000 mg/kg treatment group at test termination. There were no mortalities in any treatment group.

Conclusion: The 14-day LC50 for earthworms exposed to NOA 407405 (metabolite of thiamethoxam) was determined to be > 1000 mg /kg dry weight soil and the overall NOEC was determined to be 125 mg /kg dry weight soil based on thinning and reduced reaction observed in the three highest treatment groups.

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Section A7.5.1.2 Earthworm, acute toxicity test Annex Point IIIA XIII 3.2 Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted

EVALUATION BY RAPPORTEUR MEMBER STATE Date l I/OI/2005 Materials and Methods

Results and discussion Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies refen·ing to the (sub)heading numbers and to applicant's summmy and conclusion. Discuss ifdeviating/ram view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating/ram view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating/ram view ofrapporteur member state Remarks

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98/8 Doc IIIA 7.5.1.2 / Acute toxicity to earthworm and other non-target macro-organism section No. 03 91/414 Annex II Effects on earthworms: Acute toxicity Point addressed 8.4.1 / 03

1. Annex point(s) II A, 8.4.1 Effects on earthworms: Acute toxicity 2. Location in Dossier Section 6, 3. Authors (Year) Bryan, R.L. (1999b) Title CGA 355190: An acute toxicity study with the Earthworm in an artificial soil substrate. Report No. / Date 108-425 / 09.12.1999

Syngenta File N° 355190/0006

Source / Owner Unpublished / Syngenta Crop Protection AG 4. Testing facility Wildlife International, Ltd., 8598 Commerce Drive, Easton, Maryland 21601 5. Dates of work 21st October 1999 through to 5th November 1999 6. Test substance Company Code: CGA 355190, 7. Test method OECD-Guideline No. 207, 1984, Earthworm, Acute Toxicity Test 8. Deviations Verification of the test concentrations, stability and homogeneity in the soil were not determined. 9. GLP Performed in compliance with all the pertinent Good Laboratory Practice Standards U.S. Environmental Protection Agency 40 CFR Parts 160 and 792, 17 August 1989, OECD and Japan MAFF) with the following exception : The stability of the test substance under storage conditions at the site was not conducted in accordance with GLP.

Test System: CGA 355190, metabolite of thiamethoxam; was used a s test substance. Test organism: earthworm (Eisenia fetida); Age: mature adults average individual body weight of earthworms was 300 to 320 mg; Source: Willingham Worm Farm, Butler Georgia, U.S.A. Earthworms were dosed by uniformly mixing the test substance into an artificial soil substrate which was then dispensed into each test vessel. The moisture content of the soil was 33% approximately and the pH at preparation on day 0 was between 7.6 to 7.7. The worms were exposed to five test concentrations of 62.5, 125, 250, 500 and 1000 mg CGA 355190/kg of dry soil. Four replicate test chambers were maintained in each treatment and control group, with 10 worms in each chamber. Assessments on mortality and abnormal behaviour were made at 7 and 14 days after treatment. The worms were weighed at the beginning and end of the test and were not fed during testing. A reference toxicity test was conducted with chloroacetamide. The test units were maintained at 20-22oC under continuous light of 458-772 lux.

Findings: Acute toxicity of CGA 355190 (metabolite of thiamethoxam) to earthworms Mortality Mean live Mean Concentration weight change in Exposure LC50 of CGA 355190 [%] [mg] live period (95 % conf. weight [%] interval) [mg /kg soil] 7 d 14 d 0 d 14 d 0 - 14 d [days] [mg /kg soil] Control 0.0 0.0 320 230 -28.1 62.5 0.0 0.0 320 220 -31.3 125 0.0 0.0 310 210 -32.3 14 753 (500-1000) 250 2.5 2.5 320 210 -34.4 500 0.0 0.0 310 180 -41.9 1000 77.5 92.5 300 200 -33.3 14-d NOEC: 250 mg /kg soil

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Observations: There was no apparent aversion to the soil in any of the treatment groups on day 0, with all worms having burrowed into the soil within 75 minutes. On day 7 all worms having burrowed into the soil within 30 minutes, with the exception of of the highest test item concentration, in which aversion was noted. With one worm not having buried within 105 minutes. During the test the change in bodyweight was statistically significant (p>0.05) at the 500 mg/kg concentration tested in the comparison to the control group. No significant bodyweight differences were observed in the highest test item treatment and the control. However the statistical results for this treatment should be interpreted with caution due to the low numbers of surviving worms in this treatment. There was some thinning observed in the 500 and 1000 mg/kg treatment group at test termination. One worm from the 250 mg/kg treatment group was not found and presumed dead on day 7. However, the mortality was considered incidental.

Conclusion: The 14-day LC50 for earthworms exposed to CGA 355190 (metabolite of thiamethoxam) was determined to be 753 mg /kg dry weight soil and the overall NOEC was determined to be 250 mg /kg dry weight soil based on thinning and reduced reaction observed in the two highest treatment groups and mortality observed in the highest treatment group.

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EVALUATION BY RAPPORTEUR MEMBER STATE Date 12/01/2005 Materials and Methods

Results and discussion Conclusion

Reliability I Acceptability Remarks - COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc IIIA 7.5.1.2 / Acute toxicity to earthworm and other non-target macro-organism section No. 04 91/414 Annex II Effects on earthworms: Acute toxicity Point addressed 8.4.1 / 04

1. Annex point(s) II A, 8.4.1 Effects on earthworms: Acute toxicity 2. Location in Dossier Section 6, 3. Authors (Year) Pfeifle, V. (2000) Title Acute Toxicity of CGA 355190 to the Earthworm Eisenia fetida. Report No. / Date 2002506 / 16.10.2000

Syngenta File N° 355190/0007

Source / Owner Unpublished / Syngenta Crop Protection AG 4. Testing facility Solvias AG, Basel, Switzerland 5. Dates of work 14th September 2000 through to 29th September 2000 6. Test substance Company Code: CGA 355190, 7. Test method OECD-Guideline No. 207, 1984, Earthworm, Acute Toxicity Test Commission Directive 87/302/EEC, Official Journal of the European Communities L133, Part C, pp 95-98. 8. Deviations None

9. GLP Performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted February 2nd, 2000 [RS 813.016.5]. This Ordinance is based on the OECD Principles of Good Laboratory Practice, as revised in 1997 and adopted November 26th, 1997 by decision of the OECD Council [C(97)186/Final].

These procedures are based on the OECD Principles of Good Laboratory Practice, adopted May 12, 1981 by Decision of the OECD Council [C (81) 30 (Final)] concerning Mutual Acceptance of Data in the Assessment of Chemicals, as amended by the Council Decision- Recommendation of 2nd October 1989 concerning Compliance with Principles of Good Laboratory Practice [C (89) 87 (Final)].

Test System: CGA 355190, metabolite of thiamethoxam; was the test item. Test organism: earthworm (Eisenia fetida); Age: approximately 9 month old mature adults average replicate body weight of earthworms was 330 to 384 mg; Source: From cultures at the testing facility, orginally obtained from SARL; Moulin, Litz, France. The test units consisted of 1.5 L glass jars filled with 700 g moist soil, equivalent to 500 g dry weight (soil specifications according to OECD 207). The test item was uniformly mixed into an artificial soil substrate which was then dispensed into each test unit. The moisture content of the soil was 37.3-37.8% and the pH at preparation on day 0 was 5.8 to 6.1. The worms were exposed to five test concentrations of 95, 171, 309, 556 and 1000 mg CGA 355190/kg of dry soil, by placing them on the soil surface. Four replicate test units were maintained in each treatment and control group, with 10 worms in each unit. Assessments on the mortality and behaviour/condition were made at 7 and 14 days after treatment, at which times the soil was removed from the test units and sorted through for the worms. Following the assessment on day 7 the soil was returned to the test units and the surviving worms replaced on the soil surface. The worms were weighed at the beginning and end of the test and were not fed during testing. In addition burrowing times were noted on days 0 and 7. The results from a reference toxicity test with chloroacetamide were also reported. The test units were maintained in continuous light (mean 448 lux) at a temperature of 18.5-21.0oC

Findings:

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Acute toxicity of CGA 355190 (metabolite of thiamethoxam) to earthworms

Mortality Mean live Mean Concentration weight change in Exposure LC50 of CGA 355190 [%] [mg] live period (95 % conf. weight [%] interval) [mg /kg soil] 7 d 14 d 0 d 14 d 0 - 14 d [days] [mg /kg soil] Control 0.0 0.0 350 332 -5 95 0.0 0.0 345 308 -11 171 0.0 0.0 352 309 -12 14 > 1000 (none) 309 0.0 0.0 339 280 -18 556 0.0 0.0 345 263 -24 1000 0.0 5.0 348 225 -35 14-d NOEC: 171 mg /kg soil

Observations: In the control and all test item treatments up to and including 556 mg CGA 355190/kg dry weight soil all worms had burrowed beneath the soil surface within 15 minutes at test start and on day 7. At the highest concentration tested (1000 mg/kg dry weight soil) all worms had buried beneath the soil surface within 15 minutes of the test start. However, on day 7 worms had still not buried within 2 hours of placing them on the soil surface. No sublethal effects, such as abnormal behaviour, flaccidity or other symptoms were observed in the control and the test item concentrations of 95, 171 and 309 mg/kg dry weight soil. At 556 mg CGA 355190/kg dry weight soil no sublethal effects were noted at day 7. However, after 14 days of exposure light flaccidity and open wounds were noted. At 1000 mg CGA 355190/kg dry weight soil moderate flaccidity was noted at day 7, with open wounds and severe flaccidity being recorded by test end. A dose-dependant reduction in bodyweight was observed during the test with such reductions being statistically significantly greater than in the control, in the test item concentrations of 171 mg/kg dry weight soil and higher. Despite the statistically significant difference of the decrease in body weight in the 171 mg/kg dry weight soil treatment group, the effect can be regarded as lying within the normal biological variation of the historical control in this test system, and is therefore considered not to be biologically relevant.

Conclusion: The 14-day LC50 for earthworms exposed to CGA 355190 (metabolite of thiamethoxam) was determined to be > 1000 mg /kg dry weight soil and the overall NOEC was determined to be 171 mg /kg dry weight soil based on biologically relevant bodyweight reductions in the three highest treatment groups.

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EVALUATION BY RAPPORTEUR MEMBER STATE Date 12/01/2005 Materials and Methods

Results and discussion

Conclusion

Reliability I

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Section A7.5.1.2 Earthworm, acute toxicity test Annex Point IIIA XIII 3.2 Acceptability

Remarks

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98/8 Doc IIIA 7.5.1.2 / Acute toxicity to earthworm and other non-target macro-organism section No. 05 91/414 Annex II Effects on earthworms: Acute toxicity Point addressed 8.4.1 / 05

1. Annex point(s) II A, 8.4.1 Effects on earthworms: Acute toxicity 2. Location in Dossier Section 6, 3. Authors (Year) Porch, J.R (2000) Title CGA 322704: An acute toxicity study with the Earthworm in an artificial soil substrate. Report No. / Date 108-418A 28.03.2000

Syngenta File N° 322704/0026

Source / Owner Unpublished / Syngenta Crop Protection AG 4. Testing facility Wildlife International, Ltd., 8598 Commerce Drive, Easton, Maryland 21601 5. Dates of work 11th January 2000 through to 26th January 2000 6. Test substance Company Code: CGA 322704, 7. Test method OECD-Guideline No. 207, 1984, Earthworm, Acute Toxicity Test 8. Deviations Verification of the test concentrations, stability and homogeneity in the soil were not determined. 9. GLP Performed in compliance with all the pertinent Good Laboratory Practice Standards U.S. Environmental Protection Agency 40 CFR Parts 160 and 792, 17 August 1989, OECD and Japan MAFF) with the following exception : The stability of the test substance under storage conditions at the site was not conducted in accordance with GLP.

Test System: CGA 322704, metabolite of thiamethoxam; was used a s test substance. Test organism: earthworm (Eisenia foetida foetida); Age: mature adults average individual body weight of earthworms was 400 to 440 mg; Source: Worm Man’s Worm Farm, Monroe Twp., NJ. Earthworms were dosed by uniformly mixing the test substance into an artificial soil substrate which was then dispensed into each test vessel. The moisture content of the soil was approximately 31% of the dry weight of the soil and the pH at preparation on day 0 was between 6.7 to 6.9. The worms were exposed to six test concentrations of 1.25, 2.50, 5.00, 10.0, 20.0 and 40.0 mg CGA 322704/kg of dry soil. The test concentrations were selected based upon the results of a range finding study. Four replicate test chambers were maintained in each treatment and control group, with 10 worms in each chamber. Assessments on mortality and signs of toxicity were made at 7 and 14 days after treatment. The worms were weighed at the beginning and end of the test and were not fed during testing. A reference toxicity test was conducted with chloroacetamide.

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Findings: Acute toxicity of CGA 322704 (metabolite of thiamethoxam) to earthworms

Mortality Mean live Mean Concentration weight change in Exposure LC50 of CGA 322704 [%] [mg] live period (95 % conf. weight [%] interval) [mg /kg soil] 7 d 14 d 0 d 14 d 0 - 14 d [days] [mg /kg soil] Control 0.0 0.0 430 370 -14.0 1.25 0.0 0.0 420 330 -21.4 2.50 0.0 0.0 430 340 -20.9 14 5.93 (5.31-6.65) 5.00 5.0 30.0 400 310 -22.5 10.0 47.5 95.0 410 350 -14.6 20.0 97.5 100.0 430 - - 40.0 100.0 100.0 440 - - 14-d NOEC: 2.50 mg /kg soil

Observations: As was observed in the control group, a slight loss in average individual body weights from day 0 to day 14 is typical, since the worms were not fed during the test. There was no apparent aversion to the soil in any of the treatment groups on day 0. All worms had burrowed into the soil within one hour on Day 0. There was apparent aversion to the soil on day 7. Two worms had not burrowed in the 5.00 mg/kg group after two hours. In the 10.0 mg/kg group eight worms had not burrowed and an additional worm had partially burrowed. All worms in the two lowest treatment groups were normal in appearance and behaviour on day 14. In the 5.00 mg/kg group signs of toxicity such as necrosis, thinner in size and demonstrating reduced reaction to mechanical stimuli were observed on during the test.

Conclusion: The 14-day LC50 for earthworms exposed to CGA 322704 (metabolite of thiamethoxam) was determined to be 5.93 mg /kg dry weight soil and the overall NOEC was determined to be 2.50 mg /kg dry weight soil based on, signs of necrotic appearance, thinning and reduced reaction observed in the four highest treatment groups.

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EVALUATION BY RAPPORTEUR MEMBER STATE Date 13/01/05 Materials and Methods

Results and discussion

Conclusion

Reliability I Acceptability Remarks -

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98/8 Doc IIIA 7.5.1.2 / Acute toxicity to earthworm and other non-target macro-organism section No. 06 91/414 Annex II Effects on earthworms: Acute toxicity Point addressed 8.4.1 / 06

1. Annex point(s) II A, 8.4.1 Effects on earthworms: Acute toxicity 2. Location in Dossier Section 6, 3. Authors (Year) Gillham, A. (2002)

Title Acute toxicity (LC50) of the metabolite NOA 459602 to the Earthworm (Eisenia fetida) in an artificial soil test.

Report No., Date JW2401 / 28.02.2002

Syngenta File N° 459602/0004

Source / Owner Unpublished / Syngenta Crop Protection AG 4. Testing facility Central Science Laboratory, Sand Hutton, York, England. 5. Dates of work 12th December 2001 through to 22nd January 2002 6. Test substance Company Code: NOA 459602; 7. Test method OECD-Guideline No. 207, 1984, Earthworm, Acute Toxicity Test. Directive 87/302/EEC, Part C. 8. Deviations None

9. GLP Performed in compliance with United Kingdom Good Laboratory Practice Statutory Instrument No. 3106 December 1999. These regulations are in accordance with the OECD Principles of Good Laboratory Practice 1997 [ENV/MC/CHEM(98)17] and re believed to be consistent with the EPA (FIFRA regulations.

Test System: NOA 459602, metabolite of thiamethoxam; Test organism: earthworm (Eisenia fetida); Age: mature adults at least 2 months old with clitellum, average individual body weight of earthworms was 300 to 600 mg. Earthworms were dosed by uniformly mixing the test item into an artificial soil substrate which was then dispensed into each test vessel. The moisture content of the soil was approximately 35% of the soil dry weight and the pH at preparation on day 0 was 6 ± 0.5. The worms were exposed to two test concentrations of 100 and 1000 mg NOA 459602/kg of dry soil. Four replicate test chambers were maintained in each treatment and control group, with 10 worms in each chamber. Assessments on mortality and signs of toxicity were made at 7 and 14 days after treatment. The worms were weighed at the beginning and end of the test and were not fed during testing. A reference toxicity test was conducted with chloroacetamide at a single concentration of 50 mg/kg dry soil. The test vessels were maintained at 20.0 to 21.3oC and in continuous light of bewteen 430-525 lux.

Findings: Acute toxicity of NOA 459602 (metabolite of thiamethoxam) to earthworms

Mortality Mean live Mean Concentration of weight change in Exposur LC50 NOA 459602 [%] [mg] live e period (95 % conf. interval) weight [%] [mg /kg soil] 7 d 14 d 0 d 14 d 0 - 14 d [days] [mg /kg soil] Control 0.0 0.0 407 384 - 6 100 0.0 0.0 411 396 - 4 14 > 1000 (n.a.) 1000 0.0 0.0 413 398 - 4 14-d NOEC: 1000 mg /kg soil Chloroacetamide - 100 415 n.a. n.a. 50 mg /kg soil

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n.a. - not applicable Observations: As was observed in the control group, a slight loss in average individual body weights from day 0 to day 14 is typical, since the worms were not fed during the test. There were no mortalities or abnormal signs of behaviour (such as worms remaining on the soil surface) in the control and eirther of the two test item treatments. All worms died within 14 days of exposure to chloroacetamide at 50 mg/kg dry soil.

Conclusion: The 14-day LC50 for earthworms exposed to NOA 459602 (metabolite of thiamethoxam) was determined to be > 1000 mg /kg dry weight soil and the overall NOEC was determined to be 1000 mg /kg dry weight (the highest concentration tested).

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EVALUATION BY RAPPORTEUR MEMBER STATE Date 14/01/2005 Materials and Methods

Results and discussion Conclusion

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Section A7.5.l.2 (applicant) Earthworm, reprnduction toxicity test (applicant) Section A7.5.2.1 01 Reproduction study with other soil non-target macro- Annex Point IIIA XIII 3.2 organisms

Official 1 REFERENCE use only 1.1 Reference Batscher, R. (2000): Effects of CGA 322704 (metabolite of CGA 293343) on survival, growth, and reproduction of the earthwonn Eisenia f etida RCC Ltd., Repo1t No.: 773976 (Syngenta Project No. 2002615), 9 October 2000 (unpublished) . 1.2 Data protection Yes. 1.2.1 Data owner Syngenta Crop Protection 1.2.2 Companies with letter of access .. 1.2.3 Criteria. for data protection

2 GUIDELINES AND QUALITY ASSURANCE 2.1 Guideline study ISO 11268-2: 1998 (E) BBA Guideline Patt VI, 2-2, 1994 2.2 GLP Yes. 2.3 Deviations None.

3 METHOD

3.1 Test material CGA 322704 (metabolite of CGA 293343) 3.1.1 Lot/Batch number 3.1.2 Specification -As given in section 2 3.1.3 Purity 3.2 Refer ence Benomyl WS50% was applied in a similar, separate study at 10 mg substance -fonnula.tion/kg dry soil (equivalent to 5 mg a.s./kg dry soil). 3.3 Testing procedure 3.3.1 Preparation of the The test substance was dissolved in purified water to provide a stock test substance solution of 15 mg/L. This was diluted tenfold to provide the 0.3 mg/kg application solution, fwt her dilutions were sued to prepare the 0.18 and 0.06 mg/kg application solutions (see table A 7_ 5_ 1_2-1) 3.3.2 Application of the Ea.ch test replicate was prepared individually by the addition of 100 Ml test substance of the appropriate application solution to the individual test vessel containing 500 g soil (dty weight) . 3.3.3 Test organisms See tableA7 5 1 2-2 3.3.4 Test system See table A7 5 1 2-3 3.3.5 Test conditions See tableA7 5 1 2-4 3.3.6 Test duration 8 weeks, adults were present for the first we.eks, the second four week

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Section A7.5.l.2 (applicant) Earthworm, reprndnction toxicity test (applicant) Section A7.5.2.1 01 Reproduction study with other soil non-target macro- Annex Point IIIA XIII 3.2 organisms

was without adults 3.3.7 Test parameter Mean weight, mortality and reproduction, qualitative assessment of food consumption 3.3.8 Examination See tables A7 5 1 2-5 - 7 for details of examinations 3.3.9 Monitoring oftest No substance concentration 3 .3 .10 Statistics Williams-test, one sided smaller

4 RESULTS

4.1 Filter paper test Not perfo1med 4.1.1 Concentration Not applicable 4.1.2 Number/ Not applicable percentage of animals showing adverse effects 4.1.3 Nature of adverse Not applicable effects 4.2 Soil test 4.2.1 Initial 0.06, 0.18 and 0.3 mg/kg dty soil concentrations of test substance 4.2.2 Effect data The mortality data is presented in table A7_ 5_ 1_2-5; (mortality, weight Body weight data is presented in table A 7_ 5_ l _ 2-6; and reproduction) Reproduction data is presented in table A7 _5_1_2-7 4.2.3 Concentration I Not applicable effect curve 4.2.4 Other effects Not applicable

4.3 Results of controls 4.3. I Mortality Control mortality was 5% (2 dead wonns from 40). 4.3.2 Number/ No wonns showed any adverse effects percentage of earthwonns showing adverse effects 4.3.3 Nature of adverse Not applicable effects

4.4 Test with Perfo1med (as a separate study) reference substance

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Section A7.5.l.2 (applicant) Earthworm, reprndnction toxicity test (applicant) Section A7.5.2.1 01 Reproduction study with other soil non-target macro- Annex Point IIIA XIII 3.2 organisms

4.4.1 Concentrations 5 mg a.s./kg dry soil 4.4.2 Results Swvival rate was not significantly affected, however a significant reduction in mean body weight was obse1ved. The reproduction rate of Eisenia fetida was almost completely inhibited (0 .1% of the control value).

5 APPLICANT'S SUMMARY AND CONCLUSION 5.1 Materials and ISO 11268-2: 1998 (E) methods BBA Guideline Patt VI, 2-2, 1994 No deviations 5.2 Results and The control group mortality was 5% after 28 days exposme. Mortality discussion in the 0.06, 0.18 and 0.3 mg/kg treatments was 0, 2.5 and 0% respectively. Therefore CGA 322704 had no effect on the swvival rate of adult E. fetida up to a rate of 0.3 mg/kg dry soil. There was a tempora1y reduction in food consumption in the 0.18 and 0.3 mg/kg treatments on day 7 only. The mean body weight of the control ea1ihwomlS increased by 25% dming the exposme period. The increases in body weight in the CGA 322704 treatments were slightly lower compared to the control, 12, 19 and 19% in the 0.06, 0.18 and 0.3 mg/kg treatments respectively. However, tltis slight reduction was not statistically significant. In the control the mean number of juveniles produced per swviving adult was 17.4. The mean number of juveniles produced per swviving adult was 17 .9, 7.3 and 10.4 in the 0.06, 0.18 and 0.3 mg/kg treatments respectively. The reproduction rate was 103% of the control for the 0.06 mg/kg treatment, 42 and 60% for the 0.18 and 0.3 mg/kg treatments respectively. Reproduction was significantly reduced at the 0.18 and 0.3 mg/kg treatments (a = 0.05) 5.2.1 NOEC The highest concentration of CGA 322704 tested without toxic effects on survival, body weight and reproduction of Eiseniafetida was 0.06 mg/kg dty soil. 5.3 C onclusion The highest concentration of CGA 322704 tested without toxic effects on survival, body weight and reproduction of Eiseniafetida was 0.06 mg/kg dty soil. The validity criteria of the study were met, see table A7 5 1 2-8. 5 .3 .1 Other Conclusions None 5.3.2 Reliability 5.3.3 Deficiencies No

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Section A7.5.l.2 (applicant) Earthworm, reprndnction toxicity test (applicant) Section A7.5.2.1 01 Reproduction study with other soil non-target macro- Annex Point IIIA XIII 3.2 organisms

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted

EVALUATION BY RAPPORTEUR MEMBER STATE Date 2l/03/2006 Materials and Methods Results and discussion

Conclusion

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Table A7_5_1_2-1: Preparation of TS solution

Criteria Details Type and source of dilution water Purified water In case of the use of an organic solvent Dispersion No

Table A7_5_1_1-2: Test organisms Criteria Details Species/strain Eisenia fetida Source of the initial stock Blades Biological, Kent, TN8 7DX, United Kingdom Culturing techniques The earthworms were held for at least three weeks prior to testing. During holding they were fed with suitable food (e.g. animal manure and potatoes). Age/weight The worms used in the study were 5 – 6 months old adults with a clitellum. The age of the worms from the synchronised culture differed by not more than four weeks. Body weight of the individual worms ranged between 312 and 598 mg. Pre-treatment Three days prior to the study the worms were acclimated to the artificial soil and test temperature.

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Table A7_5_1_1-3: Test system Criteria Details Artificial soil test substrate Sphagnum moss peat – 10% Kaolinite clay – 20% Sand – 69% Calcium carbonate (CaCO3) - 0.3% Food (dried horse manure) – 1% Moisture content was adjusted to approximately 33% using purified water. Size, volume and material of test container Glass dishes with a diameter of 14 cm and a height of 7 cm. Test vessels were covered by transparent lids. Lids had a fine mesh (mesh size 0.5 mm) to allow air exchange. Amount of artificial soil (kg)/ container 500 g dry weight (soil depth of 5 – 6 cm. Nominal levels of test concentrations 0.06, 0.18 and 0.3 mg/kg dry soil Number of replicates/concentration 4 Number of earthworms/test concentration 40 Number of earthworms/container 10 Light source Room lighting Feeding 5 g of air dried horse manure was added to the test vessels when they were prepared. After the addition of the earthworms 2.5 g of food was added to each vessel, thereafter approximately 2 – 4 g was added to the adults weekly. The offspring were fed only once at the start of the 4 week period. Test performed in closed vessels due to significant No volatility of test substrate

Table A7_5_1_2-4: Test conditions

Criteria Details Test temperature Room temperature was in the range 19 to 22ºC. Temporarily the temperature was between 22 and 23 ºC due to technical problems. However, the results demonstrate that this did not affect the outcome of the study. Moisture content Soil moisture content was 33% at the start of the study and 35 – 37% at the termination of the study (8 weeks). pH pH was in the range 6.2 – 6.4 at the test start and pH was 6.2 in all treatments at the end of the study (8 weeks) Adjustment of pH No Light intensity / photoperiod Photoperiod was 16 hours light, 8 hours dark Light intensity was approximately 620 – 730 lux.

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Table A7_5_1_2-5: Mortality data Test Vessel No. of Number of Total number % mortality Substance number earthworms surviving of dead after 4 weeks Concentration exposed earthworms earthworms (nominal) after 4 weeks [mg/kg exposure artificial soil] 1 10 10 Control 2 10 9 2 5 3 10 9 4 10 10 1 10 10 0.06 2 10 10 0 0 3 10 10 4 10 10 1 10 10 0.18 2 10 9 1 2.5 3 10 10 4 10 10 1 10 10 0.3 2 10 10 0 0 3 10 10 4 10 10

Table A7_5_1_2-6: Mean body weights of earthworms at the start and end of the four week exposure period Test Vessel Day 0 Day 28 Difference Substance number Concentration (nominal) No of Mean No of Mean (mg) (%) [mg/kg earthworms weight earthworms weight artificial soil] (mg) (mg) 1 10 440 10 506 66 15 Control 2 10 456 9 614 159 35 3 10 439 9 551 112 26 4 10 494 10 609 115 23 Mean - 457 570 113 25 SD - 26 51 38 1 10 470 10 562 92 20 0.06 2 10 462 10 523 61 13 3 10 456 10 486 30 7 4 10 500 10 537 37 7 Mean - 472 527 55 12 SD - 20 32 28 1 10 461 10 503 42 9 0.18 2 10 451 9 562 112 25 3 10 455 10 545 90 20 4 10 486 10 596 110 23 Mean - 463 552 88 19 SD - 16 39 32 1 10 447 10 507 60 13 0.3 2 10 463 10 559 96 21 3 10 477 10 543 66 14 4 10 423 10 536 113 27 Mean - 453 536 84 19 SD - 23 22 25

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Table A7_5_1_2-7: Reproduction Test Vessel Juvenile worms Reproduction rate Substance number Concentration (nominal) No. per Mean ± Per Mean ± CV (%) % of [mg/kg vessel SD surviving SD control artificial soil] adult 1 152 15.2 Control 2 129 165 ± 30 14.3 17.4 ± 3.4 19.9 100 3 198 22.0 4 179 17.9 1 165 16.5 0.06 2 218 179 ± 31 21.8 17.9 ± 3.1 17.1 103 3 187 18.7 4 147 14.7 1 99 9.9 0.18 2 61 71 ± 19 6.8 7.3 ± 1.8 24.1 42* 3 62 6.2 4 63 6.3 1 110 11.0 0.3 2 131 104 ± 41 13.1 10.4 ± 4.1 38.9 60* 3 130 13.0 4 45 4.5 * - significantly different to the control (Williams Test, one sided smaller, α = 0.05)

Table A7_5_1_2-8: Validity criteria for earthworm reproduction study Criterion Fulfilled Not fulfilled Reproduction: at least 30 juveniles per test vessel in the control, yes - coefficient of variance ≤30%

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Section A7.4.3.S/Ol (applicant) Effects on any other specific, non-target 01·ganisms (flora or fauna) believed to be at 1·isk-Earthworm field study (applicant) A. 7.5.2.1 02 Annex Point IIIA XIII 2.4 Reproduction study with other soil non-target macro organisms

Official 1 REFERENCE use only 1.1 Reference Pease, G. & Webster, D. (2004) CGA322704 (A metabolite of CGA293343 (thiamethoxam)): A field study to investigate the forced effect and recovery of earthworm populations following application to a bare field site in Denmark Ecotox Ltd. Tavistock, Devon, England, Report No.: ER-04-KCB 196 (Syngenta. Project No. 2033604), 4 November 2004 (unpublished). 1.2 Data protection Yes. 1.2.1 Data owner Syngenta Crop Protection 1.2.2 Companies with letter of access •· 1.2.3 Criteria. for data protection

2 GUIDELINES AND QUALITY ASSURANCE 2.1 Guideline study BBA PartVl-2-3 (1994) and ISO 11268-3 (1999) 2.2 GLP Yes, except for weather data 2.3 Deviations None.

3 METHOD

3.1 Test material CGA 322704 (metabolite of CGA 293343) 3.1.1 Lot/Batch number 3.1.2 Specification -As given in section 2 3.1.3 Pmity 3.2 Reference •Carbenda.zim applied at a rate of 4000 g a.s./ha. substance 3.3 Testing procedure 3.3.1 Pre-treatment On 30 May 2003 a pre-study earthworm sample was conducted to assessment determine whether the site yielded sufficient numbers of earthworms per m2 (BBA 1994 and ISO 11268-3 1999 guidelines) and included appropriate representative species. Earthworm species representative of the major functional groups were present on the site at the time of the pre-treatment sampling, including Apporectodea longa, and ApOl'rectodea caliginosa, epilobous juveniles were the dominant groups in terms of numbers and biomass. Adults of other species, such as Lumbricus ten·estris and Allolobophora chlorotica, were also present. There were fewer occul1'ences of epigeic species such as Lumbricus f estivus and L. castaneus. 3.3.2 Test location and Study site - The study site was a bare earth field site in Denmark, which design had been undisturbed and untreated since 2000. The site was

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Section A7.4.3.5/0l (applican t) Effects on an y other specific., non-target 01·ganisms (flora or fauna) believed to be at 1·isk-Earthworm field study (applicant) A. 7.5.2.1 02 Annex Point IIIA XIII 2.4 Reproduction study with other soil non-target macro organisms

approximately 14 1 x 56 m, approximately 0. 75 hectare. A randomised block design of five treatments and four replicates was used. Each replicate was approximately 12 x 12 m (with an inner sampling area of 10 x 10 m) with an approximately 5 m dividing strip of grass between plots. The treatments were as follows: Control (water) 37.5 g CGA 322704/ha 75 g CGA 322704/ha 150 g CGA 322704/ha Reference substance, carbendazim 4000 g a.s./ha 3.3.3 Soil Following analysis the soil on the site was classified as a loamy sand. characterisation The mean pH was 5.7, the mean cation exchange capacity (meq/lOOg) was 7.9, mean organic matter content was 1.8% w/w and the mean water holding capacity was 11.0% w/w. The mean soil components content was sand 815, silt 95 and clay 10%. 3.3.4 Test organisms Field population of ea1ihworms 3.3.5 Application of the The application was made to the site on 16 June 2003. All treatments test substance were applied using a tractor mounted Hardi LX MB boom and nozzle sprayer with a 12 m boom and 50 cm nozzle spacing. The nozzles were Hardi ISO LD 03 110 flat fan low drift type. Water was applied to the control plots first, followed by the CGA 322704 treatments from low to high rate. The carbendazim treatment was applied last. The application volume was equivalent to 1000 L/ha. The percentage of applied volume of spray in relation to the percentage of actual volume per plot ranged from a minimum of97.9% (in the control treatment) to a maximum of 101.7% (in the 150g/ha treatment). 3.3.6 Test conditions In·igation - A pennanent Bording Mobil M5 in-igation system at the study site was used both before and after treatment application. Between 4 June 2003 and 15 June 2003 (pre-treatment), approximately 50 mm in-igation was applied to the site. A combination of 22 mm rainfall and in-igation at the site was recorded for the 3-day period following application. In the 5 day period, 8 to 13 July 2003 leading up to the first post-treatment sampling occasion approximately 34 mm in-igation was applied to the site. An electronic data monito1-ing system was installed approximately 50 m from the site, it was programmed to record minimum and maximum air and soil temperatures and daily rainfall. Soil moisture was recorded on each sampling occasion. 3.3.7 Test duration 386 days 3.3.8 Test parameter Ea1ihworm abundance and biomass 3.3.9 Sampling Ea1ihworm sampling - Sampling took place within a central 10 m x 10 m area of each plot (12 m x 12 m), using four 0.25 m2 qua.drats in each 2 plot, combined to give a. sample of l m . EarthwomlS were sampled using a digging (to a depth of approximately 30 cm) and hand-sorting method on all occasions. For a period of seven days immediately after application, surface searches were can-ied out daily and earthwonns collected from the same four 1 m2 areas per plot were identified and

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counted in the test and reference item treatments. 3.3.10 Monitoring oftest Yes. Analysis was perfonned by high pe1fo1mance liquid substance chromatography with triple quadrople mass spectrometry detection (LC concentration MS/MS). 3 .3 .11 Statistics For both abundance and biomass, totals per plot were calculated before analysis was performed. One of the assumptions of the analysis was that the variances in ea1ihworm abundance/biomass between plots in different treatment groups are equal. This assumption was appropriate for earthworm biomass but not for earthwo1m abundance, where treatment groups with larger mean counts tend to have larger variances and vice versa. Before analysis, therefore, both pre- and post treatment

abundance data were transfo1med onto a. log10 (abundance + 1) scale. This transfo1ma.tion is commonly used to coITect for the specific fo1m of heterogeneity of variance observed in abundance data. (Sokal and Rohlf, 1981). An analysis of variance, appropriate to a randomised complete block design, was performed on the transfo1med pre-treatment abundance data. and untransfonned pre-treatment biomass data. In each case, the pooled estimate of residual eITor variance obtained was used to compare each treatment to a control using a. two-sided Dunnett's t-test (Hsu, 1996) at a significant level p<0.05. An analysis of covariance, appropriate to a randomised block design, was perfonned on the transfonned post-treatment abundance data, using the transfo1med pre-treatment abundance data. as a. covariate, and on the untransfonned post-treatment biomass data, using the untransfo1med pre-treatment biomass data. as covariate. These analyses were followed by significance tests (F-tests) at the significance level p < 0.05 to detennine whether the pre-treatment abundance/biomass data. influenced the post-treatment abundance/biomass data. If the covariate was found to be significant, an analysis of covariance was selected whereas, if the covariate was not found to be significant, an analysis of variance was selected. For both abundance and biomass data, the pooled estimate of residual eITor obtained from the selected fonn of analysis was sued to compare each treatment to the control using a two-sided Dunnett's test (Hsu, 1996) at the 5% significance level.

4 RESULTS

4.1 Field study 4.1.1 Measured CGA 322704 residues in the plots applied with the lowest test rate concentrations of ranged from 18.5 to 27.6 g /ha. test substance CGA 322704 residues in the plots applied with the medium test rate ranged from 31. lto 82.9 g /ha. CGA 322704 residues in the plots applied with the highest test rate ranged from 68.0 to 114.3 g /ha. There was no contamination of the control plots. 4.1.2 Meteorological data. The meteorological data. is presented in table A 7_ 4_ 3_ 4-1.

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4.1.3 Effect data Total mean number of earthworms collected on each sampling occasion (biomass and following application of CGA322704 in the field is presented in table abundance) A7 4 3 4-2. Total mean weight (g) of earthwonns collected on each sampling occasion following application of CGA322704 in the field is presented in table A7 4 3 4-3. 4.2 Test with reference substance 4.2.1 Concentrations 4000 g a.s./ha 4.2.2 Results There were significant differences between the reference item, carbendazim, applied at 4000 g ai/ha for total numbers and biomass of earthwonns when compared with controls approximately one, four and nine months after treatment. These data confirm the validity of the study. There were no significant differences betwe.en the reference item treatment and the controls for any taxa on the final sampling occasion approximately one year after application.

5 APPLICANT'S SUMMARY AND CONCLUSION 5.1 Materials and BBA Patt V l -2-3 (1994) and ISO 11268-3 (1999). methods No deviations

5.2 Results and Results of the post-treatment surface searches showed that~ 1 % of the discussion pre-treatment sample population died on the surface during the first week after application in the test and reference item treatments. A significant reduction in abundance for ' epilobous juveniles' and 'total juveniles' was observed at the lowest test item application rate of 37 .5 g/ha approximately one month after application. Biomass for 'other epilobous juveniles' was also significantly different when compared to control at approximately one month after application. There were no significant differences in either abundance or biomass in any other groups or on any subsequent sampling occasion. There were no significant reductions in total eruihwo1m abundance on any post-treatment sampling occasion in the 75 g /ha treatment. Biomass was significantly lower in this treatment for 'total ea1thwonns' on one post-treatment sampling occasion only, approximately three months after application. There were no subsequent effects on abundance or biomass for any taxon. Abundance ofA . caliginosa was lower in the 150 g/ha treatment than in the controls on one occasion only, but biomass was significantly lower in this treatment for three taxa; A. caliginosa on one occasion only approximately nine months after application; ' other epilobous juveniles' at one and three months after application; 'total ea1ihwo1ms' at approximately three months after application. There were no effects of this treatment on either abundance or biomass on the final sampling occasion approximately one year after application. On the final srunpling occasion, approximately one yeru· after application, there were no significant differences betv.•een the controls and the test material, CGA 322704 applied at 37.5, 75or150 g /hain

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Section A7.4.3.5/0l (applican t) Effects on any other specific., non-target 01·ganisms (flora or fauna) believed to be at 1·isk-Earthworm field study (applicant) A. 7.5.2.1 02 Annex Point IIIA XIII 2.4 Reproduction study with other soil non-target macro organisms

either abundance or biomass of earthwonns. 5.3 Conclusion CGA322704, when applied at tlu·ee rates of37.5, 75 and 150 g CGA 322704/ ha showed no adverse effects on earthwonn populations for either ecological groups or individual species in samples collected one year after application of the treatments. Equivalent to a thiamethoxam maximum rate of approximately 200 g thiamethoxam/ha. 5 .3 .1 Other Conclusions None 5.3.2 Reliability 5.3.3 Deficiencies No

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Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted

EVALUATION BY RAPPORTEUR MEMBER STATE Date 21103/2006 Materials and Methods Results and discussion

Conclusion Reliability I Acceptability Remarks - COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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Table A7_4_3_4-1: Monthly rainfall, mean soil moisutre potential and minimum and maximum air and soil temperatures during the study period Date Rainfall Minimum air Maximum Minimum Maximum Mean soil (mm) temperature air soil soil moisture (°C) temperature temperature temperature potential (°C) (°C) (°C) (hPa) June 5 – 30 100.7 8.2 27.9 12.5 20.3 Pre-treat (13 2005 Jun to 02 Jul 03) 47.0 July 2003 64.4 10.3 34.6 14.7 23.1 1 MAA (07 Jul to 22 jul 03) 74.4 August 2003 64.6 4.8 34.6 12.3 23.6 * September 31.9 1.7 27.8 7.7 21.6 3 MAA (09 2003 Sep to 22 Sep 03) 91.1 October 36.2 -5.7 17.6 1.6 12.3 * 2003 November 56.3 -1.4 13.8 2.7 8.8 * 2003 December 51.8 -7.3 9.3 0.1 7.3 6 MAA 2003 (24 Nov to 10 Dec 03) 12.1 January 88.7 -11.0 6.6 -0.6 2.8 * 2004 February 49.9 -5.8 11.1 -5.8 7.1 * 2004 March 2004 47.0 -5.1 15.8 -5.1 15.8 9 MAA (09 Mar to 25 Mar 04) 17.6 April 2004 32.7 0.5 19.7 0.5 17.9 * May 2004 30.2 2.6 26.3 8.4 16.7 * June 2004 70.3 4.1 26.4 11.6 18.3 12 MAA (29 Jun to 09 Jul 04) 188.2 July 1- 9 40.1 7.1 24.2 12.9 17.9 * 2004 For the year Ttoal Minimum Maximum Minimum Maximum Mean 71.7 764.8 -11.0 34.6 -5.8 23.6 * - no soil potentiometer reading taken MAA – months after application

Table A7_4_3_4-2: Total mean number of earthworms collected on each sampling occasion following application of CGA322704 in the field Treatment Application Mean total number of earthworms collected / m2 rate Pre- 28DAT 92DAT 169DAT 274DAT 386DAT treatment Control - 120.25 76.50 85.00 72.25 88.25 63.75 37.5 g/ha 106.00 50.75 64.75 65.75 74.00 67.25 CGA322704 75 g/ha 123.25 67.00 72.00 61.50 84.00 80.25 150 g/ha 112.25 59.75 54.25 65.50 66.75 55.75 Carbendazim 4000 g ai 105.75 47.50* 40.25* 55.25 47.75* 53.75 /ha

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DAT – Days after treatment. *Significantly different from the control in Dunnett’s test, (p<0.05).

Table A7_5_1_1-3: Total mean weight (g) of earthworms collected on each sampling occasion following application of CGA322704 in the field Treatment Application Mean total number of earthworms collected / m2 rate Pre- 28DAT 92DAT 169DAT 274DAT 386DAT treatment Control - 81.63 55.81 84.42 70.10 87.48 68.75 37.5 g/ha 72.04 40.07 59.20 62.80 73.23 71.98 CGA322704 75 g/ha 86.73 47.60 60.37* 65.39 89.24 87.63 150 g/ha 74.69 49.65 53.43* 59.03 66.78 55.07 Carbendazim 4000 g ai 71.91 31.99* 40.25* 49.63 51.91* 61.30 /ha DAT – Days after treatment. *Significantly different from the control in Dunnett’s test (p<0.05).

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98/8 Doc IIIA 7.5.3.1.1 Acute oral toxicity section No. I 01 91/414 Annex II Effects on birds - Acute oral toxicity Point addressed 8.1.1 I 02

1. Annex point(s) II A, 8.1.1 Effects on Birds -Acute Oral Toxicity 2. Location in Dossier Section 6 3. Authors I Year

Title 6 CGA 293343 ACUTE ORAL TOXICITY (LD50) TO THE Report No. I Date BOBWHITE QUAIL CBG 744, 960155 / 23.04. 1996 Novartis File N° Novartis Study # 293343-46 Source I Owner Unpublished I Nova1tis Crop Protection AG

4. Testing facility

5. Dates of work December 19, 1995 to January 9, 1996 6. Test substance ISO common name thiamethoxam.

7. Test method U.S. EPA FIFRA Assessment Guidelines, Subdivision E, Section No. 71-1 (October 1982) and its revised draft (1988). 8. Deviations None noted 9. GLP The study was conducted to confonn with Good Laborat01y Practice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Patt 160, 17 August 1989; OECD, ISBN 92-64-12367-9, Paris 1982; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984. Good Laborato1y Practice, The United Kingdom Compliance Programme, Deprutment of Health and Social Security, and Department of Health, 1989. EC Council directive, 87/ 18 EEC of 18 December 1986.

Test System: Thiamethoxam technical, Thiamethoxam, dispersed in 1% methylcellulose, was administere to m 1v1 a Bo w rte qua1 , Co inus virginianus by oral intubation using a disposable syringe and a CHIO Nelaton plastic catheter. Sixty adult birds (30 males and 30 females), approximately 22 months old with a body weight range of 171-208 g at study initiation, were distributed by random draw into 6 groups of ten birds each (five males I five females), with five treatment groups and one control group. Treatment levels: 125, 250, 500, 1000 and 2000 mg/kg. The birds were examined for mortality, clinical signs, body weight gain and food consumption for a period of 14 days.

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Findings: Acute oral toxicity of thiamethoxam to the bobwhite quail

Dose Toxicological LLD, LDso NOEL [mg/kg bw] results• [mg/kg bw] [mg/kg bw] [mg/kg bw] Control 0 I 0 I IO I25 0 I 0 I IO 250 0 I 0 I IO 1000 1552 125 500 0 I 0 I IO IOOO 2 I 4 I IO 2000 7 I 9 I IO

•number of birds which died I number of birds with ove1t clinical signs I total number of birds exposed

Clinical signs: Clinical signs of toxicity were observed at 1000 and 200 mg/kg. These included subdued behaviour, unsteadiness and rnffled feathers. Some females at 2000 mg/kg did not show clinical signs of toxicity until several days after dosing. Markedly lower food consumption and large body weight losses occtmed at 1000 and 2000 mg/kg. Slight effects on both parameters were noted at 250 and 500 mg/kg.

Gross necropsy: At necropsy, clinical findings were mainly in birds from the dose group 2000 mg/kg and were generally associated with body weight loss. Red/dark coloured intestines were also observed in a few birds.

Conclusion: The acute oral LD50 of bobwhite quail exposed to thiamethoxam was determined to be 1552 mg/kg bw.

Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 16/01/2005 Materials and Methods Results and discussion Conclusion

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Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

Results and discussion Discuss if deviating from view of rapporteur member state Conclusion Discuss if deviating from view of rapporteur member state Reliability Discuss if deviating from view of rapporteur member state Acceptability Discuss if deviating from view of rapporteur member state Remarks

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98/8 Doc lllA 7 .5.3.1.1 Acute oral toxicity section No. I 02 91 /414 Annex II Effects on birds . Acute oral toxicity Point addressed 8.1.1 I 03

1. Annex point(s) II A, 8.1.1 Effects on Birds - Acute Oral Toxicity 2. Location in Dossier Section 6 3. Authors I Year

Title 7 CGA 293343 ACUTE ORAL TOXICITY (LDSO) TO THE MALLARD Report No. I Date DUCK CBG 745, 960013 I 23.04.1996 Novartis File N° Novartis Study# 293343-44 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility

5. Dates of work Janua1y 3, 1996 through January 17, 1996. 6. Test substance ISO common name thiamethoxam.

7. Test method U.S. EPA FIFRA Pesticide Assessment Guidelines, Subdivision E, Section No. 71-1 (October 1982) and draft revised guideline 71-1 dated march 1988. 8. Deviations None noted 9. GLP The study was conducted to conform with Good Laborato1y Pra.ctice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989; OECD, ISBN 92-64-12367-9, Paris 1982; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, IO August 1984. Good Laborato1y Practice, The United Kingdom Compliance Programme, Depaitment of Health and Social Sectu'ity, and Depaitment of Health, 1989. EC Cotmcil directive, 87/18 EEC of 18 December 1986.

Test System: Thiamethoxam technical, Thiamethoxam, dispersed in I% methylcellulose, was a llllll1stere to m iv1 ua M ard ducks (Anas platyrhynchos) by oral intubation using a disposable sy1inge and a CHIO Nelaton plastic catheter. Sixty adult birds (30 males and 30 females), 21 weeks old with a body weight range of 925-1375 g at study initiation, were distributed by random draw into six groups of ten birds each (five males I five females), with five treatment groups and one control group. Treatment levels: 76, 137, 247, 444 and 800 mg/kg. The birds were examined for m01tality, clinical signs, body weight gain and food consumption for a period of 14 days.

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Findings: Acute oral toxicity of thiamethoxam to the Mallard duck

Dose Toxicologic 29 LD50 NOEL a LLD [mg/kg bw] al results [mg/kg bw] [mg/kg bw] [mg/kg bw] Control 0 / 0 / 10 76 0 / 0 / 10 137 0 / 0 / 10 444 576 >76 247 0 / 10 / 10 444 4 / 10 / 10 800 7 / 10 / 10 a number of birds which died / number of birds with overt clinical signs / total number of birds exposed

Clinical signs: All treated birds vomited after dosing. Clinical signs of toxicity were observed at 247 mg/kg and above. These included subdued behaviour, unsteadiness and inability to stand. Body weight losses occurred at 444 and 800 mg/kg during the week following dosing. No food was consumed by birds from the highest treatment group of 800 mg/kg during days 1 to 3 after dosing. In all other treatment groups food consumption was generally low relative to controls. Gross necropsy: At necropsy, abnormalities were found in a few birds at 800 and 444 mg/kg found dead after dosing. These findings were mainly in the gastrointestinal tract and included colouration of intestines and watery red fluid in the abdominal cavity.

Conclusion: The acute oral LD50 of the Mallard duck exposed to thiamethoxam was determined to be 576 mg/kg bw.

29 Lowest lethal dose

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Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 16/01/2005 Materials and Methods Results and discussion

Conclusion

Reliability I A cceptability Remarks - COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state A cceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc IIIA 7.5.3.1.1 Acute oral toxicity section No. / 03 91/414 Annex II Effects on birds - Acute oral toxicity Point addressed 8.1.1 / 01

1. Annex point(s) II A, 8.1.1 Effects on Birds - Acute Oral Toxicity 2. Location in Dossier Section 6 3. Authors / Year

Title 8 ACUTE ORAL TOXICITY STUDY IN BOBWHITE QUAIL WITH CGA Report No. / Date 322704 Notox 242257, 982601 / 07.10.1998 Novartis File N° Novartis Study # 322704-17 Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility NOTOX B.V. Hambakenwetering 3, 5231 DD’s-Hertogenbosch, The Netherlands 5. Dates of work July 15, 1998 to August 12, 1998 6. Test substance

7. Test method U.S. EPA Pesticide Assessment Guidelines, FIFRA Subdivision E, Hazard Evaluation, Wildlife and Aquatic Organisms, Series 71-Avian and Mammalian Testing, § 71-1, Avian single dose oral LD50 test, October 1982 and draft revised guideline, dated March 1988. 8. Deviations None noted 9. GLP The study was conducted to conform with Good Laboratory Practice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Part 160, 17 August 1989; OECD, ISBN 92-64-12367-9, Paris 1982; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Burea, 10 August 1984. Good Laboratory Practice, The United Kingdom Compliance Programme, Department of Health and Social Security, and Department of Health, 1989. EC Council directive, 87/18 EEC of 18 December 1986.

Test System: CGA 322704 technical, CGA 322704 was administered by oral gavage, using corn oil as a vehicle, to five birds (Bobwhite quail, Colinus virginianus) of each sex at 192, 343, 617, 1111 or 2000 mg/kg body weight. A control group of five birds of each sex was dosed with vehicle (5 ml/kg body weight). Birds were observed at periodic intervals on the day of dosing and daily thereafter. Body weight was determined at day 1 (start of study), day 8, day 11 and/or day 12 (for some birds), day 15 and at death. Food consumption was measured from days 1-4, 4-8, 8-11 and 11-15. Macroscopic post-mortem examination was performed on the day of death or at termination (day 15).

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Findings: Acute toxicity of CGA 322704 (metabolite of thiamethoxam) to the Bobwhite quail

Dose Toxicological LL03o LO so NOEL results a [mg/kg bw] [mg/kg bw] [mg/kg bw] [mg/kg bw] Control 0 I 0 I IO I92 0 I 0 I IO 343 II II IO 2000 >2000 191 6I7 0 I II IO 1111 O/ I0 /10 2000 2 I IO /10

• munber ofbit·ds which died I munber of birds with ove1t clillical signs I total munber of birds exposed

Clinical signs: Clinical signs of toxicity were obse1ved from dose levels of 343 mg/kg bw onwards. These included hunched posture, lethargy, ventro-lateral recumbency, uncoordinated movement, abno1mal gait, emaciation and ptosis which were seen in most birds from the two highest dose groups (I I I I and 2000 mg/kg bw) between 3 hours and I2 days after dosing. Markedly lower food consumption and large body weight losses occlllTed at dose levels of I I I I and 2000 mg/kg bw. Slight effects on body weight gain were noted at 343 mg/kg but were not obse1ved in birds receiving 6I 7 mg/kg bw. The NOEL in this study was I9I mg/kg bw.

Gross necropsy: At necropsy, clinical findings were mainly in birds from the dose groups 6I 7, I I I I and 2000 mg/kg and were in general enlarged red spleens, emaciation and dark red livers reduced in size.

Conclusion: The acute oral LD50 of the Bobwhite quail exposed to CGA 322704 (metabolite of thiamethoxam) was dete1mined to be > 2000 mg/kg bw.

Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 16/0l/2005 Materials and Methods

30 Lowest lethal dose

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Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

Results and discussion

Conclusion

Reliability I A cceptability Remarks - COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state A cceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc lllA 7.5.3.1.2 Short term toxicity section No. I 01 91 /414 Annex II Effects on birds ·Short-term dietary toxicity Point addressed 8.1.2 / 01

1. Annex point(s) II A, 8.1.2 Effects on Birds - Short-Term Dietary Toxicity 2. Location in Dossier Section 6 3. Authors I Year

Title 9 CGA 293343 SUBACUTE DIETARY (LCSO) TO THE BOBWHITE Report No. I Date QUAIL CBG 746, 960156 I 01.05.1996 Novartis File N° Novartis Study# 293343-47 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility

5. Dates of work December 21, 1995 through December 29, 1998 6. Test substance ISO common name thiamethoxam.

7. Test method U.S. EPA FIFRA Pesticide Assessment Guidelines Subdivision E, Section No. 71-2 (October 1982), and OECD Guideline 205 ( 4 April, 1984), and ASTM Standard E857-87 (1987). 8. Deviations None noted 9. GLP The study was conducted to conform with Good Laborato1y Pra.ctice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989; OECD, ISBN 92-64-12367-9, Paris 1982; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Bm·ea, IO August 1984. Good Laborato1y Practice, The United Kingdom Compliance Programme, Depaitment of Health and Social Sectu'ity, and Depaitment of Health, 1989. EC Council directive, 87/18 EEC of 18 December 1986.

Test System: Thiamethoxam technical, Thiamethoxam was incorporated into the basal diet an o ere a .i itum to Bo w te quail ( Colinus virginianus). Ten birds per dose level, (7 days old, body weight range of 10.1- 10.3 g, unsexed), were assigned to the treatment groups by random draw. There were six treatment and two control groups. The birds received the appropriate dieta1y concentrations ad libitum for five consecutive days and were maintained on basal diet for an additional three-day obse1vation period. Treatment levels: 163, 325, 650, 1300, 2600 and 5200 mg/kg feed. Mo1talities, clinical obse1vations, body weight gain and food consumption were recorded. The mean measured concentrations of thiamethoxam in the diets fed dming the study were within I 0% of nominal values. Test diet fo1mulations were homogeneously blended and stable for a pe1iod representing the maximum time from preparation to completion of dosing.

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Findings:

Short-term dietary toxicity of thiamethoxam to the Bobwhite quail

Cose Toxicological LLC 31 LC so NOEC (mg/kg feed] results• (mg/kg feed] [mg/kg feed] [mg/kg feed] Control OI OI 20 163 0 I 0 I IO 325 0 I 0 I IO 650 0 I 0 I IO > 5200 > 5200 1300 1300 0 I 0 I IO 2600 0 I 0 I IO 5200 0 I 0 I IO a munber of birds which died I munber of bit·ds with ove1t clinical signs I total munber of birds exposed

Observations: There were no mo1talities, overt symptoms of toxicity or behavioural abno1malities at any test substance concentration. Group mean body weight gain was reduced at 2600 and 5200 mg/kg feed relative to controls. Although food consumption was variable between the groups, there were no treatment-related trends apparent. Due to the reduction in body weight gain at 2600 and 5200 mg/kg feed, the no obse1ved effect level was considered to be 1300 mg/kg feed. The sho1t-te1m dieta1y LC50 of the Bobwhite quail was dete1mined to be > 5200 mg/kg feed. From day 0 to day 5 the mean bodyweight of birds in the highest treatment group was 15.85 g/bird and the mean food consumption in this treatment during this period was 5.88 g food/bird/day. Therefore the LC50 can alternatively be expressed as > 1.929 g thiamethoxam/kg b.w./day taking into account the actual consumption and bodyweights recorded in this study. Gross necropsy: No treatment related abno1malities were detected.

Conclusion: The sho1t-te1m dietaiy LC50 of the Bobwhite quail exposed to thiamethoxam was dete1mined to be > 5200 mg/kg feed, equivalent to an LD50 of or > 1.929 g thiamethoxam/kg b.w./day.

Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 16/0l/2005 Materials and Methods

31 Lowest lethal concentration

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Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

Results and discussion

Conclusion

Reliability I A cceptability Remarks - COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state A cceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc lllA 7 .5.3.1.2 Short term toxicity section No. I 02 91 /414 Annex II Effects on birds · Short-term dietary toxicity Point addressed 8.1.2 I 02

1. Annex point(s) II A, 8.1.2 Effects on Birds - Short-Term Dietary Toxicity 2. Location in Dossier Section 6 3. Authors I Year

Title 10 CGA 293343 SUBACUTE DIETARY TOXICITY (LC50) TO THE Report No. I Date MALLARD DUCK CBG 747, 960199 I 23.04.1996 Novartis File N° Novartis Study# 293343-45 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility

5. Dates of work December 21, 1995 through December 29, 1995 6. Test substance ISO common name thiamethoxam.

7. Test method U.S. EPA FIFRA Pesticide Assessment Guidelines, Subdivision E, Section No. 71-2 (October 1982), and OECD Guideline 205 (4 August 1984), and ASTM Standard E857-87 (1987). 8. Deviations None noted 9. GLP The study was conducted to conform with Good Laborato1y Pra.ctice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160, 17 August 1989; OECD, ISBN 92-64-12367-9, Paris 1982; and Japan MAFF, 59 NohSan, Notification No. 3850, Agricultural Production Bm·ea, IO August 1984. Good Laborato1y Practice, The United Kingdom Compliance Programme, Depaitment of Health and Social Secm-ity, and Depaitment of Health, 1989. EC Council directive, 87/18 EEC of 18 December 1986.

Test System: Thiamethoxam technical, Thiamethoxam was incorporated into the basal diet an o ere a i itu.m to M ard duck (Anas platyrhynchos). Ten birds per dose level, (6 days old, body weight range of 83 - 84 g, unsexed), were assigned to the treatment groups by random draw. There were six treatment and two control groups. The birds received the appropriate dieta1y concentrations ad libitu.m for five consecutive days and were maintained on basal diet for an additional three-day obse1vation period. Treatment levels: 163, 325, 650, 1300, 2600 and 5200 mg/kg feed. Mo1talities, clinical obse1vations, body weight gain and food consumption were recorded. The mean measured concentrations of thiamethoxam in the diets fed dming the study were within I 0% of nominal values. Test diet fo1mulations were homogeneously blended and stable for a pe1iod representing the maximum time from preparation to completion of dosing.

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Findings:

Short-term dietary toxicity of thiamethoxam to the Mallard duck

Dose Toxicological LLC32 LC so NOEC [mg/kg feed] results• [mg/kg feed] [mg/kg feed] [mg/kg feed] Control OI OI 20 163 0 I 0 I IO 325 0 I 0 I IO 650 0 I 0 I IO > 5200 > 5200 163 1300 0 I 0 I IO 2600 0 I 0 I IO 5200 0 I 0 I IO a munber of bit·ds which died I munber of bit·ds with ove1t clinical signs I total munber of birds exposed

Observations: There were no mo1talities, overt symptoms of toxicity or behavioural abno1malities at any test substance concentration. A treatment related reduction in group mean body weight gain was noted at 1300 mg/kg feed and above. A slight reduction was also noted at 325 and 650 mg/kg feed. In the post-treatment period a compensato1y gain in body weight had occuned in these dose groups. There was also a reduction in feed consumption in the three highest dose groups during the treatment period which had improved by the end of the study. Due to the reduction in body weight gain and food consumption noted at 1300 mg/kg feed and above and the slight reduction in body weight gain at 325 and 650 mg/kg feed, the no obse1ved effect level was considered to be I 63 mg/kg feed. From day 0 to day 5 the mean bodyweight of birds in the highest treatment group was I50.5 g/bird and the mean food consumption in this treatment during this pe1iod was 34 g food/bird/day. Therefore the LC50 can alternatively be expressed as > l. I 75 g thiamethoxam/kg b.w./day taking into account the actual consumption and bodyweights recorded in this study. Gross necropsy: No abno1malities were detected.

Conclusion: The sho1t-te1m dieta1y LC50 of the Mallard duck exposed to thiamethoxam was dete1mined to be > 5200 mg/kg feed, equivalent to an LD50 of> 1.175 g thiamethoxam/kg b.w./day.

Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 17/0l/2005 Materials and Methods Results and discussion

32 Lowest lethal concentration

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Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

Conclusion :

Reliability Acceptability Remarks

COMMENTS FROM ... (specify) Date Give date of comments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summary and conclusion. Discuss if deviating from view of rapporteur member state Results and discussion Discuss if deviating from view of rapporteur member state Conclusion Discuss if deviating from view of rapporteur member state Reliability Discuss if deviating from view of rapporteur member state Acceptability Discuss if deviating from view of rapporteur member state Remarks

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98/8 Doc lllA 7.5.3.1.3 Effects on reproduction section No. I 01 91 /414 Annex II Effects on birds • Subchronic toxicity and reproduction Point addressed 8.1.3 / 02

1. Annex point(s) II A, 8.1.3 Effects on Birds - Subchronic Toxicity and Reproduction 2. Location in Dossier Section 6 3. Authors I Year

Title 11 THE REPRODUCTIVE TOXICITY TEST OF CGA-293343 TECHNICAL WITH THE NORTHERN BOBWHITE (COLINUS VIRGINAJ\1US) Report No. I Date 029518 / 09.07.1998 Novartis File N° Novartis Study# 293343-653 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility 5. Dates of work September 5, 1996 through March 28, 1997 6. Test substance ISO common name thiamethoxam.

7. Test method US EPA Pesticide Assessment Guidelines, Subdivision E, Hazard Evaluation: Wildlife and Aquatic Organisms (EPA, 1982), and in ASTM Standard Practice for Conducting Avian Reproduction Test, DRAFT No. 9, 1983. 8. Deviations none 9. GLP The study was conducted to conform with Good Laborato1y Practice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160.

Test System: Thiamethoxam technical, adult bobwhites (Colinus virginianus), approac mg t eu irst r and 1 da ; wei t between 168.7-237. l g for males and females; . One hundred and forty-four birdS, me u mg ·ee ose groups an one contro group were employed in the study. Each treatment and the control group consisted of 18 replicates, one male and one female per replicate. Birds were offered test diet containing 100, 300 and 900 mg thiamethoxam/ kg feed over a period of 23 weeks. After 13 weeks of treatment, first eggs were set for incubation and observed until 14 days post-hatch. Exposure of the parental generation continued for 10 weeks of egg production. Adults were observed daily for mortality, abnormal behaviour and signs of toxicity. Reproductive parameters such as the number of eggs laid, number of eggs cracked, egg shell thickness, embryonic viability and chick survival were assessed. The mean measured concentrations of thiamethoxam in test diet formulations analysed during the study were 95 .08 ± 13.3 for the 100 mg/kg feed level; 309.89 ± 28.4 for the 300 mg/kg feed level and 946.42 ± 129.8 for the 900 mg/kg feed dose level confirming the accuracy of preparation. Storage stability of the test substance under frozen and ambient storage conditions was also performed. No significant loss ofthiamethoxam occurTed over the storage periods.

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Findings: Reproductive toxicity of thiamethoxam to the Northen bobwhite Reproductive parameters Control 100 mg / 300 mg 900 mg / kg feed kg feed kg feed Total eggs laid in each group 990 1139 918 876 Eggs laid / per hen 61.8 63.3 61.2 51.5 Eggs laid / per day per hen 0.87 0.89 0.86 0.73 Eggs cracked 9 10 9 8 Eggs cracked / eggs laid 0.009 0.009 0.010 0.009 Mean egg shell thickness (mm) 0.209 0.207 0.208 0.210 Eggs set 901 1040 835 786 Fertile eggs 815 986 787 706 Fertile eggs/ eggs set 0.90 0.95 0.94 0.90 Viable embryos 794 972 777 690 Viable embryos / fertile eggs 0.97 0.99 0.99 0.99 Hatchlings 755 933 745 650 Hatchlings / viable embryos 0.95 0.96 0.96 0.94 14-Day old survivors 429 501 497 412 14-Day old survivors / of hatchlings 0.57 0.54 0.67 0.63 Hatchlings / eggs set 0.84 0.90 0.89 0.83 14-Day old survivors / eggs set 0.48 0.48 0.60 0.52 14-Day old survivors/hen 26.8 27.8 33.13 24.2 Mean chick body weight at hatching (g) 6.22 6.39 6.21 6.16 Mean chick body weight at 14 days (g) 22.07 20.94 22.84 22.62 NOEL 900 mg/kg feed

Observations: There were six mortalities in the adult quail population during the treatment period including birds from all dose groups and the control group. It was considered that these mortalities were not treatment related and all surviving birds were noted to be normal in appearance and behaviour with a few exceptions. No overt signs of treatment-related toxicity were observed in the study. Bodyweight and food consumption in the treatment groups were not significantly different from the control group. Post-mortem examination of adult birds revealed no treatment-related abnormalities and all findings are regarded as typical in a large group of birds maintained on an ad libitum feed. There were no significant differences on the reproductive parameters detected at any level of treatment when statistically compared against the control group.

Conclusion: The reproductive no-observed-effect concentration during this study was determined to be 900 mg/ kg feed (equivalent to 74.0 mg a.i./kg b.w./day).

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Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 16/01/2005 Materials and Methods Results and discussion Conclusion

Reliability I A cceptability Remarks - COMMENTS FROM ... (specify) Date Give date ofcomments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state A cceptability Discuss ifdeviating from view ofrapporteur member state Remarks

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98/8 Doc lllA 7 .5.3.1.3 Effects on reproduction section No. I 02 91 /414 Annex II Effects on birds · Subchronic toxicity and reproduction Point addressed 8.1.3 / 01

1. Annex point(s) II A, 8.1.3 Effects on Birds - Subchronic Toxicity and Reproduction 2. Location in Dossier Section 6 3. Authors I Year

Title 12 THE REPRODUCTIVE TOXICITY TEST OF CGA-293343 TECHNICAL WITH THE MALLARD DUCK (ANAS PLATYRHYNCHOS) Report No. 029710 I 09.11.1998 Novartis File N° Novartis Study # 293343-889 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility 5. Dates of work March 3, 1998 through September 8, 1998 6. Test substance ISO common name thiamethoxam. Com an Code: CGA 293343 tech.,

7. Test method US EPA Pesticide Assessment Guidelines, Subdivision E, Hazard Evaluation: Wildlife and Aquatic Organisms (EPA, 1982), and in ASTM Standard Practice for Conducting Avian Reproduction Test, DRAFT No. 9, 1983. 8. Deviations None 9. GLP The study was conducted to conform with Good Laborato1y Practice Standards as published by the U.S. Environmental Protection Agency, Office of Pesticide Programs in 40 CFR Pait 160.

Test System: CGA 293343 technical, adult Mallard Duck (Anas platyrhynchos), approac gt eu u st ree and 1 da ; wei ht between 937.7 - 1022.6 for males and females; . One hundred and twenty-e1g t u s, me u mg t ·ee ose groups an one contro group were employed in the study. Each treatment and the control group consisted of 16 replicates, one male and one female per replicate. Birds were offered test diet containing 100, 300 and 900 mg CGA 293343/ kg feed over a period of 189 days (27 weeks). After 11 weeks and 2 days of treatment, first eggs were set for incubation and obse1ved until 14 days post-hatch. Exposure of the parental generation continued for 10 weeks of egg production. Adults were obse1ved daily for mo1tality, abno1m al behaviour and signs of toxicity. Reproductive parameters such as the number of eggs laid, number of eggs cracked, egg shell thickness, emb1yonic viability and chick smvival were assessed. The mean measured concentrations of CGA 293343 in test diet fo1mulations analysed dming the study were 101.82 ± 19.47 for the 100 mg/kg feed level; 307.14 ± 34.33 for the 300 mg/kg feed level and 908.75 ± 73.72 for the 900 mg/kg feed dose level confnming the accuracy of preparation. Storage stability of the test substance under frozen and ambient storage conditions was also perfo1med. No significant loss of CGA 293343 occmTed between the storage pe1iods and feed mixing inte1vals; at the 100 and 900 mg/kg feed dose levels, a 10.5% and 6.7% decrease in concentrations were obse1ved by day 21 when feed mix was again prepared.

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Findings: Reproductive toxicity of thiamethoxam to the Mallard duck Reproductive parameters Control 100 mg / 300 mg / 900 mg / kg feed kg feed kg feed Number of Replicates (productive pairs) 16 14 12 16 Total eggs laid in each group 718 580 558 551 Eggs laid / per hen 44.88 41.43 46.50 34.44 Eggs laid / per day per hen 0.64 0.59 0.66 0.49 Eggs cracked 44 39 25 22 Eggs cracked / eggs laid 0.06 0.07 0.04 0.04 Mean egg shell thickness (mm) 0.353 0.357 0.361 0.353 Eggs set 616 492 480 479 Fertile eggs 567 453 457 427 Fertile eggs/ eggs set 0.92 0.92 0.95 0.89 Viable embryos 536 412 421 378 Viable embryos / fertile eggs 0.95 0.91 0.92 0.89 Hatchlings 458 334 361 324 Hatchlings / viable embryos 0.85 0.81 0.86 0.86 14-Day old survivors 420 320 345 306 14-Day old survivors / of hatchlings 0.92 0.96 0.96 0.94 Hatchlings / eggs set 0.74 0.68 0.75 0.68 14-Day old survivors / eggs set 0.68 0.65 0.72 0.64 14-Day old survivors/hen 26.25 22.86 28.75 19.13 Mean chick body weight at hatching (g) 36.4 35.7 35.3 35.2 Mean chick body weight at 14 days (g) 189.7 225.3 222.6 229.5 NOEL 300 mg/kg feed

Observations: There was only one mortality in the adult mallard population (female, 100 mg/kg feed test group) during the treatment period including birds from all dose groups and the control group. The mortality was not treatment related and the majority of birds were noted to be normal in appearance and behaviour with a few exceptions. No overt signs of treatment- related toxicity were observed in the study. There were five females that did not lay eggs at any time during this study, one in the 100 mg/kg feed group and four in the 300 mg/kg feed group. All hens in the 900 mg/kg feed group laid eggs. Since there was no indication of a dose- response relationship, the occurrence of non-layers in the 100 and 300 mg/kg feed groups was considered not to be related to the test substance. Food consumption in the treatment groups was not significantly different from the control group. Post-mortem examination of adult birds revealed no treatment-related abnormalities and all findings are regarded as typical in a large group of birds maintained on ad libitum feed. There were no significant differences in the reproductive parameters detected at any level of treatment when statistically compared against the control group. Subtle yet statistically significant decreases in adult body weight were detected in males from the 900 ppm test groups as compared to the controls in the last two weighing intervals. At termination, the average adult male body weights for 0, 100, 300, and 900 mg/kg feed test groups were 1225.7, 1197.4, 1113.6, and 1109.3 grams, respectively.

Conclusion: The reproductive no-observed-effect concentration during this study was determined to be 300 mg/kg feed (equivalent to 34.9 mg a.i./kg b.w./day).

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Section 7.5.3.1.1 Acute oral toxicity on birds Annex Point IIIA XIII 1.1 Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 19/01/2005 Materials and Methods Results and discussion

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98/8 Doc lllA 7.5.4.1 Acute toxicity to honey bees and other beneficial arthropods, for example section No. 01 predators 91 /414 Annex II Bees: Acute toxicity Point addressed 8.3.1.1 01

1. Annex point(s) II A, 8.3.1.1 Bees: Acute Toxicity 2. Location in Dossier Section 6 3. Authors I Year Kleiner, R. (1995)

Title 13 TESTING TOXICITY TO HONEYBEE -APIS MELLIFERA L. (LABORATORY) ACCORDING TO EPPO GUIDELINE NO. 170: CGA-293343 Report No. I Date 951048045 I 25.10.1995 Novartis File N° Novartis Study# 293343-18 Source I Owner Unpublished I Novartis Crop Protection AG

4. Testing facility BIOCHEM GmbH Karlsrnhe, Labor Cmillersdorf, Am Wieseneck 7, D-04451 , ClUUle1-sdo1f, Gemiany 5. Dates of work September 20, 1995 through September 27, 1995 6. Test substance ISO common name thiamethoxam.

7. Test method EPPO Guideline No. 170 8. Deviations None 9. GLP Study was conducted according to the Principles of Good Laboratory Practice (GLP) Chemikaliengesetz., July 1994, Anhang 1).

Test System: Thiamethoxam technical, est species: Apis mellifera L. (Hymenoptera: Apidae ; Gottscheina, Germany. Oral test: Bees were dosed by group feeding using a 50% sucrose solution. Nominal concentrations were used. The treatments were as follows: Six test doses of thiamethoxam (ranging from 0.002- 0.02 µg ai/bee), one control dose (50% sucrose-water solution) and six doses of a toxic standard (dimethoate 400 g/L EC, ranging from 0.20 to 0.40 µg ai/bee). Test duration: 48 hours. Experimental design: 3 replicates/treatment, 10 bees/ replicate. Biological observations on mortality and behaviour were made at 24 and 48 hours after application. Contact test: Bees were dosed by topical application. Nominal concentrations were used. The test substance was dissolved in acetone. Bees received six test doses of thiamethoxam ranging from 0.005 to 0.05 µg ai/bee, one canier control dose (acetone) and six doses of a toxic standard (dimethoate 400 g/L EC, ranging from 0.0313 to 1.0 µg ai/bee). Test duration: 48h. Experimental design: 3 replicates/ treatment, 10 bees/replicate. Biological observations on mortality and behaviour were made at 24 and 48 hours after application. Findings: Acute toxicity of CGA 293343 via oral exposure in the laboratory Mortality following Exposure LDso Dose level oral exposure period [µg/bee] [%)

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24h 48 h [hours] [µg/bee] control sucrose 0 0 0.002 0 0 0.004 43 43 24 0.005 thiamethoxam 0.008 83 83 0.12 87 87 48 0.005 0.016 97 97 0.02 97 97 0.20 3 3 0.24 3 10 24 0.374 Toxic standard 0.28 13 23 0.32 40 50 48 0.335 0.36 30 53 0.40 67 77

Acute toxicity of CGA 293343 via contact exposure in the laboratory

Mortality following Exposure LDso Dose level contact exposure period [µg/bee] [%) 24h 48 h [hours] [µg/bee] control sucrose 0 0 0.005 0 0 O.Ql 0 7 24 0.027 thiamethoxam 0.02 17 23 0.03 67 70 48 0.024 0.04 87 87 0.05 100 100 0.0313 0 0 0.0625 0 7 Toxic standard 0.125 0 13 24 0.363 0.25 7 30 0.5 90 90 48 0.260 1.0 100 100

Observations: Affected bees showed restlessness, initation, uncontrollable motions and dorsal position before dying. Twenty-four hours and 48 hours after application the smviving bees exhibited no behavioural anomalies. In the reference treatment apathy, uncontrollable motions and dorsal position of affected bees could be obse1ved before dying.

Conclusion: The acute 48-hour LD50 of Apis mellifera exposed to thiamethoxam via oral exposure in the laborato1y was dete1mined to be 0.005 µg/bee. The acute 48-hour LD50 ofApis mellifera exposed to thiamethoxam via contact exposure in the laborato1y was dete1mined to be 0.024 µg/bee.

Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted

EVALUATION BY RAPPORTEUR MEMBER STATE

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Evaluation by Competent Authorities Date 20/01/2005 Materials and Methods Results and discussion Conclusion

Reliability Acceptability Remarks

COMMENTS FROM Date Give date ofthe comments submitted Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state

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98/8 Doc IIIA 7.5.4.1 / Acute toxicity to honey bees and other beneficial arthropods, for example section No. 02 predators 91/414 Annex II Bees: Acute toxicity Point addressed 8.3.1.1 / 02

1. Annex point(s) II A, 8.3.1.1 Bees: Acute Toxicity 2. Location in Dossier Section 6 3. Authors / Year Nengel, S. (1997)

Title 14 ASSESSMENT OF SIDE EFFECTS OF CGA 322704 TO THE HONEYBEE, APIS MELLIFERA L. IN THE LABORATORY

Report No. / Date 97071/01-BLEU, 972512 / 14.07.1997

Novartis File N° Novartis Study # 322704-11

Source / Owner Unpublished / Novartis Crop Protection AG

4. Testing facility GAB Biotechnology GmbH, Niefern, Germany 5. Dates of work April 22, 1997 through April 24, 1997 6. Test substance Company Code: CGA 322704,

7. Test method EPPO Guideline No. 170 8. Deviations None 9. GLP Study was conducted according to the Principles of Good Laboratory Practice (GLP) Chemikaliengesetz, July 1994, Anhang 1). The OECD Principles of Good Laboratory Practice

Test System: CGA 322704 technical, . Test species: Apis mellifera L. (Hymenoptera: Apidae); Source: beekeeper (B. Deger) in Baden- Würtemberg, Germany. Age: approximately 22- 32 days old. Oral test: Bees were dosed by group feeding using a 50% sucrose solution. Nominal concentrations were used. Five test doses of the metabolite of thiamethoxam (ranging from 0.0016-0.0625 µg CGA 322704/bee), one control dose (50% sucrose-water solution) and four doses of a toxic standard ‘Rogor’ (dimethoate 400 g/L EC, ranging from 0.0675 to 0.15 µg a.i./bee) were tested. Test duration: 48 hours. For each treatment five replicate groups of 10 bees were tested. Contact test: Bees were dosed by topical application. Nominal concentrations were used. The test item was dissolved in water in combination with a surface-tension-lowering substance (“Citowett” at 1 mL/L water). Bees received five test doses of the metabolite of thiamethoxam ranging from 0.0016 to 0.0625 µg CGA 322704/bee, one carrier control dose (water in combination with a surfactant-tension-lowering substance: ‘Citowett’ 1 mL/L) and four doses of a toxic standard (dimethoate 400 g/L EC, ranging from 0.08-0.20 µg ai/bee). Test duration: 48h. For each treatment, five replicate groups of 10 bees were tested. Foloowing both oral and contact exposure observations on mortality and behaviour were made at 24 and 48 hours after dosing. Findings:

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Acute toxicity of the metabolite CGA 322704 via oral exposure

Mortality following Exposure LDso Dose level oral exposure period (95 % conf. lua/bee] 1%1 interval) actual nominal intake 24h 48 h [hours] [µg/bee] control sucrose 0 0 0 0.0016 0.0015 0 0 0.004 0.0044 0 4.0 CGA 322704 0.01 0.0114 28.0 28.0 48 0.0168 0.025 0.0222 64.0 66.0 (0.0139- 0.0625 0.0481 100 100 0.0203) 0.675 0.0791 0 4.0 Dimethoate 0.084 0.0956 10.0 24.0 48 0. 1263 0. 105 0.1208 36.0 42.0 (0. 1181- 0.1351 ) 0.15 0.1647 66.0 80.0

Acute toxicity of the metabolite CGA 322704 via contact exposure in the laboratory

Mortality following Exposure LDso Dose level contact exposure period (95 % conf. (µg/bee) [%] interval)

nominal 24h 48 h [hours] [µg/bee] control Can'ier a 0 0 0.00 16 0 0 0.004 4.0 4.0 CGA 322704 0.01 4.0 10.0 48 0.0275 0.025 32.0 38.0 (0.0226- 0.0625 88.0 88.0 0.0335) 0.08 0 0 Dimethoate 0.12 4.0 4.0 48 0. 1644 0.16 28.0 42.0 (0. 1573- 0. 1718) 0.20 80.0 88.0 •water in combination with a swfactant-tension-lowering substance: ' Citowett' 1 mL/L

Observations: In regard to the behaviour the treated bees did not differ from the controls at any time during the test. Results obtained for the toxic standard dimethoate where within the accepted range indicating the validity of this study.

Conclusion: The acute 48-hour LD50 of Apis mellifera exposed to CGA 322704 via oral exposure in the laborato1y was detennined to be 0.0168 µg/bee. The acute 48-hour LD50 of Apis mellifera exposed to thiamethoxam via contact exposure in the labora.to1y was detennined to be 0.0275 µg/bee

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE

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Evaluation by Competent Authorities Date 20/01/2005 Materials and Methods Results and discussion Conclusion

Reliability Acceptability Remarks

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98/8 Doc lllA 7 .5.4.1 Acute toxicity to honey bees and other beneficial arthropods, for example section No. 03 predators 91 /414 Annex II Point addressed 8.3.1.1

1. Annex point(s) II A, 8.3.1.1 Non target arthropods 2. Location in Dossier Section 6 3. Authors I Year Grimm, C. (1998a) Title Acute Toxicity Test of CGA 293343 to the predato1y groWld beetle Poecilus cupreus L. (Coleoptera: Carabidae). Report No. I Date 983663, November 2, 1998 Syngentra File N° 293343/0876 Source I Owner Unpublished I Nova1tis Crop Protection AG 4. Testing facility Nova1t is Crop Protection AG, Basel, Switzerland 5. Dates of work September 4, 1998 through September 21, 1998

6. Test substance ~93343 tech.,

7. Test method Heimbach, U. (1992): Laboratory method to test effects of pesticides on Poecilus cupreus (Coleoptera, Carabidae). IOBC/WPRS Bulletin 1992/XV/3:103-109 .. 8. Deviations The test item was applied as a seed dressing and not sprayed on the sand surface 9. GLP Yes - ce1t ified laboratory

~m : CGA 293343 --Test species: Poecilus cupreus L. Co eoptera, Cara i ae ; Age: 4-5 wee o a u t beetles; Source: BTL Bio-Test Labor GmbH, Birkenallee 19, D-18184, Sagerheide, Ge1many. Adult beetles, housed in plastic boxes (175 cm2 surface area) filled with approximately 250 g qua1tz sand moistened to 70% of its maximum water holding capacity, were exposed by adding treated summer wheat seeds (Triticum aestivum var. Greina) to the test Wlits (placed on the sand surface) nominally dressed with A-9700 B at a rate of 70 g a.i./100 kg seed. The sowing rate was equivalent to 200 kg wheat seeds/ha. The treatments were as follows: one test unit each containing 10 wheat seeds dressed with the test item, with a deionised water control and with a toxic standard (methyl-parathion WP 40) at a dressing rate equivalent to 160 g a.i./100 kg seeds, respectively. Expe1imental design: six beetles (three male and three female)/replicate, five replicates/treatment. Test duration: 14 days. At test initiation, and on test days 2, 4, 7 and 10 the beetles were fed one perforated fly pupae, Calliphora spp., per smviving beetle. Biological obse1vations on mo1tality and behaviour were recorded 2, 4 6 hours and at 1, 2, 4, 7, 10 and 14 days after treatment. In addition, food consumption (fly pupae) was recorded at 2, 4, 7, 10 and 14 days after treatment. At these times it was also recorded whether the beetles appeared to have been feeding on the treated seeds. The test units were maintained at a temperature of l 7.0- 20.00C, a relative humidity range of 65-96% Wlder a 16 hour light:8 hour dark regime with a light intensity of 900-1100 lux.

Findings: Acute toxicity of treated wheat seeds on P. cupreus adults Wlder laborato1y conditions

Effects on Mortality and Food Consumption

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Treatment 14· day mean Average number of fly pupae mortality consumed per evaluation [%] interval fpupae/beetlel Control 0 0.57 CRU ISER"" 350 FS 70 a ai/100 ka seeds 100 0.50 Toxic standard 100 n.a. n.a. - not assessed Observations: No abno1mal behaviour of the beetles was obse1ved in the control. The first signs of detrimental changes in behaviour of the beetles treated with the test item were obse1ved 4 hours after exposure and 2 hours after exposure in the toxic standard treatment. After 4 days of exposure in the test item treatment and after 2 days of exposure in the toxic standard treatment all beetles were found dead. The mean food consumption per beetle per assessment inte1val was 0.57 pupae and 0.50 pupae in the control and test item treatment, respectively. However, it should be noted that due to the total m01tality in the test item treatment by 4 days, the consumption value for this treatment is based only upon the feeding during first 2 days of exposure. No feeding consumption data were obtained for the toxic standard treatment as there were no smviving beetles at the first feeding evalution.

Conclusion: After 4 days of exposure to wheat seeds nominally dressed with 200 mL )/kg seed (equivalent to 70 g a.i./100 kg seed) and a sowing rate a, I 00 % m01tality of P. cupreus adults occmTed under laborato1y conditions.

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 25/01/2004 Materials and Methods

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Evaluation by Competent Authorities Results and discussion

Conclusion

Reliability Acceptability Remarks

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98/8 Doc IIIA 7.5.4.1 / Acute toxicity to honey bees and other beneficial arthropods, for example section No. 04 predators 91/414 Annex II Point addressed 8.3.1.1

1. Annex point(s) II A, 8.3.1.1 Non target arthropods 2. Location in Dossier Section 6 3. Authors / Year Grimm, C. (1998b) Title Acute toxicity of CGA 293343 to the rove beetle Aleochara bilineata Gyll. (Coleoptera Staphylinidae) Report No. / Date 983664, November 3, 1998 Syngentra File N° 293343/0877 Source / Owner Unpublished / Novartis Crop Protection AG 4. Testing facility Novartis Crop Protection AG, Basel, Switzerland 5. Dates of work September 4,1998through September 25, 1998 6. Test substance Company Code: CGA 293343 tech.,

7. Test method Samsoe-Petersen, L. (1992): Laboratory method for testing side-effects of pesticides on the rove beetle Aleochara bilineata-adults. IOBC Bulletin 1992/XV/3: 82-88. 8. Deviations The test item was applied as a seed dressing and not sprayed on the sand surface 9. GLP Yes – certified laboratory

Test System: CGA 293343 Test species: Aleochara bilineata gyllenhal (Coleoptera Staphylinidae); Age: 7 to 12-day old female beetles; Source: De Groene Vlieg, Nieuwe Tonge, Netherlands. Adult beetles, housed in plastic test units (surface area of 18.86 cm2), containing a layer of quartz sand (36 g moist sand/test unit), were exposed by adding treated summer wheat seeds (Triticum aestivum var. Greina) to the test units (placed on the sand surface) nominally dressed with A- 9700 B at a rate of 70 g a.i./100 kg seed. The sowing rate was equivalent to 200 kg wheat seeds/ha. The treatments were as follows: one test unit each containing 1 wheat seed dressed with the test item, with a deionised water control and with a toxic standard (methyl-parathion WP 40) at a dressing rate equivalent to 160 g a.i./100 kg seeds, respectively. Experimental design: ten replicates per treatment each with 1 Aleochara bilineata female. Test duration: 14 days (4 days of exposure, followed by a 10 day egg viability phase). Mortality, behaviour and food consumption (number of fly eggs of Protophormia terraenovae) of the beetles were recorded on test day 1, 2, 3 and 4. The number of Aleochara bilineata eggs laid until test day 4 were detemined following their separation from the sand substrate, and the number of hatched larvae were recorded over the following 10 days, whilst being maintained on moist filter paper in Petri dishes. The test units were maintained at a temperature of 17.0-21.0oC, a relative humidity range of 65-90% under a 16 hour light: 8 hour dark regime with a light intensity of 180-400 lux.

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Findings: Acute toxicity of treated wheat seeds on A. bilineata females under laborato1y con 1tions

Effects on Mortality and Food Effects on Reproduction Consumption Treatment Cumulative Average number Average Hatching success Mortality on day 4 of onion fly eggs number of of the eggs laid consumed day 0- eggs laid during the 4 day [%] day 4 during the 4 exposure phase [eggs/beetle] day exposure [%] phase reaas /beetle1 Control 0 217.7 53.6 93.4 CRUISER® 350 FS 90 46.0 0.0 n.a. b 70 a a.i./1 00 ka seeds Toxic standard 100 n.a. a n.a. b n.a. b • n.a.: not applicable due to 100% mortality b n.a.: not applicable as no eggs were laid Obse1vations: No abno1mal behaviour of the beetles was obse1ved in the control. Exposure of rove beetles to - treated wheat seeds had severe effects on the mo1tality, feeding rate ~On days 1 and 3 of exposure the number of consumed onion fly eggs was significantly reduced in the test item treatmenillrou compared to the control group. No eggs were produced in the - treatment group compared to 53 .6 eggs per beetle in the conti·ol g~ 93.4% o them hatching. Since no eggs were laid following test item treatment, no hatching rate could therefore be dete1mined.

Conclusion: After 4 days of exposure to wheat seeds nominally dressed with 200 mL kg seed (equivalent to 70 g a.i./100 kg seed) and a sowmg rate o 200 g see s a, 90 % mo1tality of A. bilineata adults occun ed under laborato1y conditions. The food consumption during this period was reduced and no eggs were oviposited. Resulting in an R-value (based on reproduction) of 0.

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 26/01/2004

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Evaluation by Competent Authorities Materials and Methods

Results and discussion

Conclusion

Reliability Acceptability Remarks

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Evaluation by Competent Authorities Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state

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98/8 Doc IIIA 7.5.4.1 / Acute toxicity to honey bees and other beneficial arthropods, for example section No. 05 predators 91/414 Annex II Point addressed 8.3.1.1

1. Annex point(s) II A, 8.3.1.1 Non target arthropods 2. Location in Dossier Section 6 3. Authors / Year Reber, B. (2000) Title Acute Toxicity Test of CGA 293343 to larvae of the predatory ground beetle Poecilus cupreus L. (Coleoptera: Carabidae). Report No. / Date 2003631, November 7, 2000 Syngentra File N° 293343/1336 Source / Owner Unpublished / Novartis Crop Protection AG 4. Testing facility Novartis Crop Protection AG, Basel, Switzerland 5. Dates of work September 26, 2000 through October 24, 2000 6. Test substance Company Code: CGA 293343 tech.,

7. Test method Heimbach, U. (1998): Testing the effects of plant protection products on larvae of the carabid beetle Poecilus cupreus (Coleoptera, Carabidae) in the laboratory, method and results. IOBC/WPRS Vol. 21(6): 21-28. 8. Deviations The test item was applied as a seed dressing and not sprayed on the sand surface 9. GLP Yes – certified laboratory

Test System: CGA 293343 Test species: Poecilus cupreus L. (Coleoptera, Carabidae); Age: 24-48 hour old larvae; Source: BioChem Agrar, D-04451, Cunnersdorf, Germany. Beetle larvae introduced into glass tubes (2.2 cm diameter, 7 cm high) containing 5 cm depth of LUFA 2.1 soil adjusted to 30% of its water holding capacity. Just prior to beetle introduction a single pea seed Pisum sativum and one half of a thawed pupae of Calliphora spp. (food) was placed on the soil surface. Each tube was covered by a lid with holes of 2mm diameter to allow ventillation. The pea seeds introduced for the test item treatment group had been dressed with 150 mL /100 kg seed (equivalent to 52.5 g a.i./100 kg seed). Those in the toxic standard treatment group had been dressed with 400 g Methylparathion WP 40 (containing 40% methylparathion)/100 kg seed. Seeds introduced into the control test units had been treated with deionised water. The sowing density in all treatment groups was equivalent to 7143 kg seeds/ha (resulting in an application rate of 3750 g a.i./ha). This application rate is approximately 60 times and 29 times the maximum application rates of thiamethoxam resulting for the proposed uses of in cereals and peas, respectively. Experimental design: 1 larvae/replicate, 40 replicates/treatment. Test duration 14 days. Biological observations were made 3 times/week during which the mortality was determined. At these times the surviving larvae were fed with half of a thawed pupa of Calliphora spp. placed on the soil surface and the old food was removed. The test units were maintained in darkness at a temperature of 19.5- 21.0oC and 68-82% relative humidity during the test.

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Findings: Effects of treated pea seeds on la1vae of P. cupreus under extended la Treatment mean

100

100 • all treatments received pea seeds sown at a density equivalent to 7143 kg seeds/ha Obse1vations: After 5 days of exposure, all la1vae in the test item treatment and in the toxic standard treatment were found dead on the soil surface. Conclusion: After 5 days of exposure to pea seeds dressed with 150 mL - - kg seed (equivalent to 52.5 g a.i./100 kg seed) and a s owing~ ~ m01tality of P. cup reus laivae occuned under extended laborato1y conditions. It should be noted that this exagerated sowing density of 7143 kg seeds/ha resulted in a a ve1y unrealistic application rate of 3750 g a.i./ha. This application rate is approximately 60 times and 29 times the maximum application rates of thiamethoxam resulting for the proposed uses of CRUISER® 350 FS in cereals and peas, respectively

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 27/0l/2005

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Evaluation by Competent Authorities Materials and Methods

Results and discussion

Conclusion

Reliability Acceptability

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Evaluation by Competent Authorities Remarks

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98/8 Doc lllA 7 .5.4.1 Acute toxicity to honey bees and other beneficial arthropods, for example section No. 06 predators 91 /414 Annex II Point addressed 8.3.1.1

1. Annex point(s) II A, 8.3.1.1 Non target arthropods 2. Location in Dossier Section 6 3. Authors I Year Candolfi, M.P., (1998a) Title Acute Toxicity Test of CGA 293343 to la1vae of the predato1y groWld beetle Poecilus cupreus L. (Coleoptera: Carabidae). Report No. I Date 9837720ctober 28, 1998 Syngentra File N° 293343/0797 Source I Owner Unpublished I Nova1tis Crop Protection AG 4. Testing facility Nova1tis Crop Protection AG, Basel, Switzerland 5. Dates of work June 8, 1998through June 22, 1998 6. Test substance

7. Test method BARRETT, K.L., GRANDY, N., HARRISON, E.G., HASSAN, S. ANDOOMEN, P. (eds.) 1994: Guidance Document on Regulato1y Testing Procedures for Pesticides with Non-Target Arthropods. From the ESCORT Workshop (Emopean Standard Characteristics of Beneficial Regulatory Testing); Wageningen, Holland, 28 - 30 March, 1994. Dohmen et al., (1998): Testing side effects of pesticides on carabid beetles. Draft Ring-Testing Guideline. A joint initiative ofEPPO, BART, IOBC and COMET. Heimbach, U., Buchs, U. & Abel, Ch. (1992): A semi-field method close to field conditions to test effects of pesticides on Poecilus cupreus (Coleoptera, Carabidae). IOBC/WPRS Bulletin XV/3: 159-165. 8. Deviations The test item was applied as a seed dressing and not sprayed on the sand surface 9. GLP Yes - ce1tified laboratory

Test System: Test species: Poeci us cupreus L. Co eoptera, Cara 1 e ; Age: a u t eet es 23 ays a er emerging from hibernation); Source: BTL, Sagerheide, Ge1many. Adult beetles, housed in exposure units (50 cm square metal frames, which enclosed an area of 0.25 m2 and approximately 25 cm deep). The units were sllllken 10-15 cm into the soil with approximately 10 cm protmding. The soil at the field site in Stein (No1them Switzerland) had the following characte1istics; 58.29% sand, 17.33% clay and 24.38% silt, the organic carbon was 1.96% and pH was 7.14. A Summer wheat (cultivar Greina) seed density of 200 kg seeds/ha was used. The seeds were equally distiibuted in rows (distance between rows 7 cm and 2.5 cm distance between seeds in the row) at a depth of approximately 1-2 cm. The units were covered with a large mesh netting to avoid disturbance by birds or other large animals yet minimizing the influence of the microclimate. Treatments were as follows: a.i.1100 kg seeds, equivalent to 140 g a.i./ha with a seed ens1ty o 200 g w eat see s a, a conti·ol (seeds treated with tap water) and a toxic standard (ME 605), which was applied on bran (which se1ved as bait) at the following dosage based on 1 ha. 450 g fo1mulated product + 24 L water + 50 kg bran. The bait was spread on to the soil after planting the seeds (the seeds having been treated only with tap water). Experimental design: ten beetles (five males and five females)/replicate, four

- 631 - RMS: Spain Thiamethoxam Doc III-A replicates/treatment. Test duration: 14 days. At test initiation, and on test days 2, 4, 7 and 10 the beetles were fed 10 fly pupae, Calliphora spp. fixed on cardboard (2 pieces of cardboard per replicate each with five pupae). Biological observations on mortality and behaviour were recorded at 1-3 hours after beetle introduction and thereafter at 1, 2, 4, 7, 10 and 14 days after test initiation. In addition, food consumption (fly pupae) was recorded on 2, 4, 7, 10 and 14 days after treatment.

Findings: Effects of treated wheat seeds on P. cupreus adults under semi- field cond1t1ons

Effects on Mortality and Food Consumption Treatment Mortality Average number of fly pupae [%] consumed per beetle per dav Control 7.5 0.27 CRUISER® WS 70 25.0 0.26 70 a a.i./100 ka seeds Toxic standard 82.5 0.28

Over the 14 da s of exposure a significant effect on mortality was observed in treatment when compared to the control, however the 18.9 Yc. At test termination sublethal effects on beetle behaviour treatment; 33.3% of the smviving s owed co-ordination problems, whilst behaviour. No overall effect on feeding treatment when control.

Conclusion: applied at a rate of 70 g a.i./ 100 kg seeds (equivalent to 140 g a.1. a w1 a see ens1ty o 200 kg wheat seeds/ha) resulted in a conected mortality of 18.9% after 14 days of exposure under semi-field conditions and 33.3% of the beetles demonstrated co-ordination problems.

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 27/01/2005

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Evaluation by Competent Authorities Materials and Methods

Results and discussion

Conclusion

Reliability Acceptability Remarks

COMMENTS FROM Date Give date ofthe comments submitted

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Evaluation by Competent Authorities Materials and Methods Discuss additional relevant discrepancies referring to the (sub)heading numbers and to applicant's summaiy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and discussion Discuss ifdeviating from view ofrapporteur member state Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state

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98/8 Doc lllA 7.5.4.1 Acute toxicity to honey bees and other beneficial arthropods, for example section No. 07 predators 91 /414 Annex II Point addressed 8.3.1.1

1. Annex point(s) II A, 8.3.1.1 Non target arthropods 2. Location in Dossier Section 6 3. Authors I Year Candolfi, M.P., (1998b) Title Toxicity o to Aleochara bilineata Gyll. (Coleoptera, Staphylinidae) under semi-field conditions. Report No. I Date 983771, November 17, 1998 Syngentra File N° 293343/0842 Source I Owner Unpublished I Nova1tis Crop Protection AG 4. Testing facility Nova1tis Crop Protection AG, Basel, Switzerland 5. Dates of work June 26, 1998 through August 31, 1998 6. Test substance

7. Test method BARRETT, K.L., GRANDY, N., HARRISON, E.G., HASSAN, S. ANDOOMEN, P. (eds.) 1994: Guidance Document on Regulato1y Testing Procedures for Pesticides with Non-Target Arthropods. From the ESCORT Workshop (European Standard Characteristics of Beneficial Regulatory Testing); Wageningen, Holland, 28 - 30 March, 1994. MORETH, L., NATON, E. (1992): Richtlinie zur P1iifung der Nebenwirkung von Pflanzenschutzmitteln auf Aleochara bilineata Gyll. (Col., Staphilinidae) (Halbfreilandp1iifung). Bulletin IOBC/WPRS 1992/XV/3; 152-158 Naton, E. (1988): Richtlinie zur P1iifung der Nehenwirlmng von Pflanzenschutzmitteln auf Aleochara bilineata Gyll. (Col., Staphilinidae) (erweite1ter Laborversuch). Bulletin IOBC/WPRS 1988/XI/4; 119-126. 8. Deviations The test item was applied as a seed dressing and not sprayed on the sand surface 9. GLP Yes - ce1tified laboratory

The reproductive perfo1mance (parasitismii!f onion fl Delia antiqua pupae) by adult Aleochara bilineata exposed to treated seeds (treated with - under semi-field conditions with rain protection by automatically closing, UV-pe1mea e plas~mg was assessed. The exposure units were plastic containers (57 cm x 37 cm, approximately 21 cm high) containing an approximately 11-12 cm layer of Speyer 2.1 soil. The moisture content of the soil was maintained at approximately 35-40% of the maximum water holding capacity. A seed (Summer wheat) density of 4.218 g seeds/unit was calculated based on 200 kg seeds/ha. The seeds were equally distributed in rows (distance betv.•een rows 7 cm) and planting approximately 1 cm deep. The units were covered with a fine mesh netting to avoid predation and test insect escape. On days 0, 1, 3, 6, 8, 10, 13, 17 and 20 the beetles were fed with thawed Chironomus sp. larvae. On days 6, 13 and 20, approximately 5000 D. antiqua pupae per replicate (unit) were added to the exposure units. The fly pupae being buried in 3 rows (1-3 cm deep). The second and third introductions of fly pupae were placed in new rows, each beside the previous rows. On day 27 after exposure, the rows were opened and all onion fly pupae were carefully removed and set up under laboratory conditions.

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The units used to assess parasitism rate consisted of a plexiglass tube (diameter 24 cm, height 25 cm) sealed on the top with a netting and on the bottom with an aluminium plate. Inside the tube a sieve positioned on top of a glass dish (diameter 18.5 cm, height 9 cm) was used to hold the parasitised pupae. The emerging adult beetles fell through the sieve into the dish, where they were collected and counted to detennine the number of offspring per replicate. The parasitised pupae remained in the units until all the adult A. bilineata emerged.

Treatments were as follows: 70 g a.i.1100 kg seeds, equivalent to 140 g a.i./ha with a seed density o 200 g w eat see s a, a tap water control and a toxic standard (Curaten GR 5, at a rate of 1 g/m funow). Experimental design: 200 beetles (I 00 males and 100 females)/replicate, four replicates per treatment. Test duration: Exposure time; 27 days under field conditions. Reproduction time; 35 days under laborato1y conditions. Findings: treated wheat seeds on A. bilineatafecundity under

Percentage fly Reductio n in the Treatment Treatment Emergence pupae reproductio n level rate parasitised by A. compared to contro l bilineata [total no. of beetles [%) emerged from the fly u ae 3184 21 .2 1064 7.1 66.6 3 0.0 99.9 • seed treatment b For each row of seeds planted, 0.57 g Curatell' GR 5 was spread in the soil near the seeds

treatment

Conclusion: applied at a rate of 70 g a.i./ 100 kg seeds (equivalent to 140 g a.i. a wit a see ens1ty o 200 kg wheat seeds/ha) resulted in a 66.6 % reduction ofA. bilineata fecundity compared to the control under semi-field conditions.

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 28/01/2005

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Evaluation by Competent Authorities Materials and Methods

Results and discussion

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Evaluation by Competent Authorities Conclusion

Reliability Acceptability Remarks

COMMENTS FROM Date Give date ofthe comments submitted Materials Discuss additional relevant discrepancies referring to the (sub)heading numbers and to and Methods applicant's summmy and conclusion. Discuss ifdeviating from view ofrapporteur member state Results and Discuss ifdeviating from view ofrapporteur member state discussion Conclusion Discuss ifdeviating from view ofrapporteur member state Reliability Discuss ifdeviating from view ofrapporteur member state Acceptability Discuss ifdeviating from view ofrapporteur member state

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98/8 Doc IIIA 7.5.4.1 / Acute toxicity to honey bees and other beneficial arthropods, for example section No. 08 predators 91/414 Annex II Point addressed 8.3.1.1

1. Annex point(s) II A, 8.3.1.1 Non target arthropods 2. Location in Dossier Section 6 3. Authors / Year Ruggle and Bolsinger., (1998) Title Biological activity of metabolitesof Thiamthoxam CGA293343 on insects Report No. / Date and mites. Syngentra File N° October 14, 1998 Source / Owner 293343/0894 Unpublished / Novartis Crop Protection AG 4. Testing facility Novartis Crop Protection AG, Basel, Switzerland 5. Dates of work August 1, 1995 through July 21, 1998 6. Test substance Company Code: CGA 293343 tech.,

7. Test method Internal screening method 8. Deviations n.a. 9. GLP No

Test System: the metabolites were tested for biological activity in standard laboratory screening assays as follows: Spodoptera littoralis: Contact/feeding activity: Cotton leaf discs on agar were sprayed with test solution and infested with L-1 larvae. Systemic activity: Corn seedlings were directly treated in test solution. Six days after introduction, the leaves were cut and transferred into petri dishes with moist filter paper and infested with L-1 larvae. Diabrotica balteata: Treatment of corn seedlings in a petri dish and infestation with L-3 larvae Heliothis virescens: Fresh eggs (0-24 hours old) were placed on filter paper in a Petri dish. On top went artificial diet that was treated with the test solution. Aphis craccivora: The test solution was sprayed on pea seedlings that were infested with a mixed population. Myzus persicae: The test solution was sprayed on pea seedlings that were infested with a mixed population. Nilaparvata lugens: Rice seedlings were sprayed with test solution and infested with N-3 nymphs. Tetranychus urticae: Bean leaf discs on agar in a Petri dish were infested with mixed population. Findings: Mortality (%) of metabolites of CGA 293343 on insects and mite bioassays at 100 ppm.

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Test Species Test method CGA355190 NOA404617 NOA407475 CGA 322704 Aphis craccivora contact 0 0 0 100 Myzus persicae systemic 0 0 0 100 Spodopfera littoratis feelina/contact 0 0 0 100 Spodopfera littoratis svtemic 0 0 0 100 Heliothis virescens eaa hatch 0 0 0 100 L-1 mortalitv 0 0 0 n.a. L-1 effect 0 0 0 n.a Diabrotica balteata feeding/contac 0 0 0 100 t Ni/aparvata tuaens N-3 mortality 0 0 0 0 F-1 reduction -- 0 100 Tetranvchus urticae eaa hatch 0 0 0 0 larval mortality 0 0 0 0 adult mortalitv 0 0 0 0

Conclusion: CGA355190, NOA 404617 and NOA 407475 show no biological activity in the bioassays while CGA 322704 is ve1y active except of Tetranychus urticae.

Evaluation by Competent Authorities

EVALUATION BY RAPPORTEUR MEMBER STATE Date 28/01/2005 Materials and Methods

Results and discussion Conclusion

Reliability Acceptability

Remarks

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Official 1 REFERENCE use only 1.1 Reference Bader, U. (2001): The effects ofCGA 322704 (metabolite of thiamethoxam (CGA 293343)) on the decomposition of organic material in a field litterbag test. Syngenta Crop Protection AG, Basel, Switzerland, unpublished report No. 2002619. 1.2 Data protection Yes. 1.2.1 Data owner Syngenta Crop Protection. 1.2 .2 Companies with letter of access .. 1.2.3 Criteria for data protection

2 GUIDELINES AND QUALITY ASSURANCE 2.1 Guideline study None available; but based upon the minutes of a meeting in Februaiy 2000 organised by the BBA Biologische Bundesanstalt fur Land- und Forstwit1schaft Bundesrepublik Deutschland) on the requirement of data according to Council Directive 91/414/EEC, annnex III, point 10.6.2. 2.2 GLP Yes (ce1tified laboratory), with the exception of the detennination of soil prope1ties and measurement of weather conditions. 2.3 Deviations None.

3 METHOD 3.1 Test material CGA 322704 (metabolite of thiamethoxam). 3.1.1 Lot/Batch number 3.1.2 Purity - 3.2 Reference Benlate® WP 50 (benomyl). substance - 3.3 Testing procedure Within the field 12 plots (each of 4 x 4 m) were marked in an area of approximately 25 x 25 m. With each plot 2-3 meters apait. There were three treatment groups tested (control, test item and reference item), with 4 plots randomly assigned to each treatment. In each plot 36-40 litterbags were randonly distributed, separated from one another by at least 44 cm and 50 cm from the plot margin. Each bag was 12 x 12 cm wide, made from synthetic netting with a mesh size of 6-8 mm, three sides of the bag were sown together. Into each bag was placed 2.95-5.02 g ( diy weight) of the wheat straw, cut into 5-10 cm pieces. The individual weights of the bags were recorded before test start. The fourth side of the bags were then closed with steel clamps. With the litterbags placed on the meadow surface (mown 7 days before application) within their respective plots a single application was made to the litterbags on 14th June 2000 at volumes equivalent to 500 L/ha with a backpack sprayer, with measures taken to avoid di·ift to neighbouring plots. The test item CGA 322704 was applied at a

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Section A 7.5.1.2(01) (applicant) Acute toxicity test on earthworms or other soil non-target organisms (applican t) Section 7.5.6 Effects on other terrestrial non-target organisms Annex Point IIIA XIII.3.2 concentration of 141.4 mg/L (equivalent to 70.7 g/ha). The reference item Benlate® WP 50 was applied at 4 kg a.i.(benomyl)/ha and the control was treated with tap water. After the spray residues had dried (1 how) , the litterbags were bmied approximately 5 cm deep amongst the roots of the meadow grass within the respective plots. In such a way that the bags lay horizontally amongst the roots of the meadow grass, covered by the grass. The bags were sampled, 0, 33, 92, 155 and 275 days after the application. On each sampling occasion, eight randomly selected bags were collected from each plot (32 bags/treatment/sampling time). The degradation of the straw was measmed based on the detennination of the ash free dry weight of the initial straw used and of the straw recovered at the sampling days.

4 RESULTS 4.1 Soil test See 5.2. 4.2 Test with See 5.2. reference substance

5 APPLICANT'S SUMMARY AND CONCLUSION 5.1 Materials and As above. methods 5.2 Results and The results of all treatments showed sirnilar variations of the detennined discussion degradation values (relative standard deviation = 9.8%, range 4.8-21.7% with only 2 values > 15%). The relative standard deviations over the 4 treated replicates of each group were 9.2%, 9.3% and 11.0% for the control, the reference item and the test item, respectively. The difference of decomposition of the reference item to the control was statistically highly significant (p=0.001) at day 33, day 155 and day 275. The difference of the test item to the control was statistically significant (p=0.05) at day 33 and day 275. On days 33 and 275 the deviation from the control in the CGA 322704 treated group was 8.5% and 4.3%, respectively. In line with current guidance, since the deviation at test end is less than 10%, no unacceptable effects on litter decomposition are noted for this treatment group.

The comparisons of the percentage degradation (see table) show no relevant difference between the control and CGA 322704, but a slower degradation following the reference item treatment in the first 92 days of the test and a faster degradation in the following 183 days. The average degradation rates again show a sirnilar comse of degradation in the control and CGA 322704 treatment, but a lower degradation rate of the reference item dw·ing the period 0-33 days, an almost equal rate in the next period (33-92 days) and a much faster rate in the third observation interval (92-155 days). Dming the final period of the test (155-275 days) similar degradation rates were again observed. 5.3 Conclusion No relevant differences from the control in the degradation of organic material in the field were observed dming the 275 day test period following the application of CGA 322704 (metabolite of thiamethoxam at 70.7 g/ha). 5.3. l Reliability 1.

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Evaluation by Competent Authorities Use separate "evaluation boxes" to provide transparency as to the comments and views submitted

EVALUATION BY RAPPORTEUR MEMBER STATE Date 7-03-2006 Materials and Methods Results and discussion Conclusion Reliability Acceptability

Remarks

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Organic matter litter decomposition after treatment with CGA 322704 (metabolite of thiamethoxam) Degradation n days after application (%) Deviationa Deviationa Day Deviationa Day Deviationa Treatment Day 33 Day 92 (%) (%) 155 (%) 275 (%) Control 48.5 - 73.8 - 78.7 - 85.6 - CGA 44.4 8.5* 74.2 -0.5 76.4 2.9 81.9 4.3* 322704 Toxic 38.5 20.6* 69.8 5.4 88.7 -12.7* 90.9 -6.2* reference

a Deviation (%) = deviation of treatment relative to control * statistically significantly different from the control at p=0.05 Negative values for effect indicate a faster decomposition compared to the control.

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ANNEX TO DOC IIIA section 7 ECOTOXICOLOGICAL PROFILE INCLUDING ENVIRONMENTAL FATE AND BEHAVIOUR. Statement Regarding the Use of Endpoints from the Thiamethoxam Microcosm Study and CGA 322704 Litterbag Study in the Evaluation of Thiamethoxam as a Wood Preservative PT-8.

For submission to EU Rapporteur Member State (Spain)

2nd March 2006 Dr. Adrian Gray Ecological Sciences 1. Introduction The following studies have recently been submitted to the Ministerio de Sanidad y Consumo, Madrid. • Outdoor microcosm study to assess the effects of thiamethoxam on aquatic organisms (Ashwell et al., 2003). • Effects of CGA 322704 (metabolite of thiamethoxam) on the litter decomposition of organic material in a litterbag test (Bader, 2001). The purpose of the studies was to define toxicity endpoints for thiamethoxam and its metabolite, respectively. Once defined, such endpoints along with other relevant study information, can be used to evaluate the risks for different use patterns, including foliar and seed dressing plant protection products and biocides. The intention of this statement is adapt these endpoints, such that they may be usefully employed to evaluate the relevant potential risks following the use of thiamethoxam as a wood preservative PT-8.

2. Thiamethoxam microcosm study 2.1 Overall summary of microcosm study (Ashwell et al., 2003) Section A 7.4.3.(01) Effects on aquatic organisms – further studies Annex Point IIIA XIII.3

1 REFERENCE 1.1 Reference Ashwell, J., Dark, R. Emburey, S. (2003) Thiamethoxam Outdoor Microcosm Study to Assess Effects on Aquatic Organisms. Syngenta, Jealott’s Hill International Research Centre, Bracknell, Berkshire, UK. Unpublished Report no. RJ3379B (Syngenta File no. CGA293343/1851). Study dates. 15th April – 19th September 2002.

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The study was conducted to a dose response design, with a single application of each of the five nominal treatments (100, 30, 10, 3, and 1 μg ai/L). The application was made by direct addition and thorough mixing to ensure rapid distribution throughout the water column. The initial concentrations of thiamethoxam in the microcosms were measured at between 81 and 130% of nominal (mean 99%) and so it was considered that the target treatment levels had been achieved. As a result the study endpoints were reported in relation to nominal application concentrations of thiamethoxam: • No Observed Effect Concentration (NOECcommmunity): 30 μg ai/L • No Observed Ecollogically Acceptable Effect Concentration (NOEAEC): 100 μg ai/L – Based only on an isolated effect on Chironomidae, observed only in the emergence traps samples on day 15, and was not observed at any subsequent sampling point. Thiamethoxam concentrations then declined rapidy following application. A summary of the analytical determinations of thiamethoxam concentrations in these treatments in the water column are presented in the table below.

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Table 1: Analytical determinations of thiamethoxam in the water column of dosed microcosms Microcosm Nominal Measured thiamethoxam concentration (μg ai/L) identity treatmentconcentration 0 DAT 3 DAT 7 DAT 14 DAT 21 DAT (μg ai/L) M01 30 28 4.3

2.2 Biocide Exposure Scenarios in Comparison to Microcosm Study Results The relevant scenarios to be considered for the wood preservative PT-8 use of thiamethoxam are as follows and will be considered in turn: a) Potential surface water contamination following leaching during storage. b) Surface water contamination following potential emissions from treatment plant to STP. c) Potential surface water contamination following leaching from Noise Barriers (in Service). 2.2.1 Potential surface water contamination following leaching during storage. Syngenta proposes that this risk of exposure is mitigated by restricting the storage of industrial treated timber to hard standing (preventing the direct losses to soil and to drains). Thus the exposure to surface waters via this potential route will be negligible and need not be considered further. 2.2.2 Surface water contamination following potential emissions from treatment plant to STP. In comparing the results from the microcosm study with the predicted exposure concentrations from this emission scenario one should consider the following points: • The rapid dissipation of thiamethoxam from the surface water in the microcosm study is at least in part expected to have been mediated by plants present within the ponds. Such influences leading to rapid dissipation, explains why effects seen in laboratory studies were not observed in this study conducted under more natural conditions. • The onset of effects of the neurotoxin neonicotinoid thiamethoxam are clearly expected to occur soon after exposure. Therefore the risk to aquatic organisms from thiamethoxam should focus on the acute risk. • Thiamethoxam is not expected to bioaccumulate.

The scenario assumes 3% of the total amount used/day is emitted to a STP (influent). In the case of thiamethoxam a conservative assumption is made of no reduction in concentration due to dissipation in the STP. The resultant PEC local in surface waters assumes a 10-fold dilution of the STP effluent upon reaching a moving surface water body. The table below summarises the calculated PEClocal aquatic (at time 1) for the proposed application methods.

Table 2: Thiamethoxam PECSW for following potential emissions from wood treatment plant via STP

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Thiamethoxam Scenario PEC STP (influent) PEC local aquatic (time 1) application rate (μg ai/L) (μg ai/L) 15 g/m3 Noise barrier 1.26 0.126 0.1 g/m2 Noise barrier 0.34 0.034 0.1 g/m2 Dipping 22.5 2.25 15 g/m3 Vacuum pressure 6.75 0.675 15 g/m3 Double Vacuum 3.375 0.375

The worst-case PEC local aquatic (time 1) is 2.25 μg ai/L (see Table 2). The effluent from a STP is expected to be emitted to a moving surface water body. Therefore an acculation of thiamethoxam concentrations above this worst-case predicted concentration in the surface waters is not expected, since in addition to the expected dissipation/degradation in natural systems, the exchange/dilution in water is likely to result in daily total water exchange. This worst-case concentration is 13 times lower than the microcosm NOEC and 44.4 times lower than the NOEAEC. Another way to look at this is by assuming a concentration of 2.25 μg ai/L in the surface water, but in addition also assume there is no dissipation/degradation and dilution, but rather continued similar daily effluent from the STP to the surface water (i.e. highly unrealistic). For the worst- case exposure scenario of dipping treatments it would take 13 days of inputs from the STP for the PECSW to approach the microcosm NOEC. The dissipation of thiamethoxam from the water columns in the microcosm study summarised in Table 1 are presented graphically in the figures below. In order to compare the level of exposure over the first 21-days of this study with a similar period relating to biocides emissions, for each microcosm study treatment the Area Under the Curves (AUC) were calculated using the following formulae (Trapezium rule):

AUC (between day1 and dayn) n 1 ≈ ( − )( + ) ∑ +1i dayday i +1i concconc i =1i 2 Where: conci = concentration on day i

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Figure 1: Measured concentrations (means) of thiamethoxam in the water column of microcosms nominally dosed at 30 μg ai/L (NOEC treatment)

10 Thiamethoxam Conc. (ug/L Conc. Thiamethoxam

0 0 7 14 21 DAT

Figure 2: Measured concentrations (means) of thiamethoxam in the water column of microcosms nominally dosed at 100 μg ai/L (NOEAEC treatment)

100

) 80 -1

20 Thiamethoxam Conc. (ugThiamethoxam L 0 0 7 14 21 DAT

The AUC for the NOEC and NOEAEC treatments were determined to be 100.49 and 291.85 μg ai/L x d, respectively. A similar approach was followed assuming constant exposure for 21 days to a concentration of 2.25 μg ai/L. I.e unrealistic constant exposure to this peak concentration. This is presented graphically below.

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Figure 3: Predicted concentrations of thiamethoxam in surface waters assuming constant exposure to the worst-case peak concentration for 21-days

3 )

-1 2.5 g L μ 2

1.5

0.5 Thiamethoxam Conc. ( Conc. Thiamethoxam

0 0 7 14 21 DAT

The AUC in this instance was determined to be 47.25 μg ai/L x d. This is half that calculated for the microcosm NOEC treatment. This indicates that the overall exposure to pelagic aquatic organisms in the microcosm study NOEC treatment group was approximately twice that following a 21-day rather unrealistic worst-case scenario following wood preservative treatment with thiamethoxam. Only after 45 days of constant exposure to this peak thiamethoxam PECSW following wood preservative use would the overall concentration to aquatic organisms in the surface waters be expected to exceed that of the microcosm NOEC treatment. As described previously in reality exposure in surface waters will be greatly reduced due to water exchange/dilution, dissipation and degradation in natural moving water systems. In addition, the effluent from the STP assumes a worst-case of no dissipation. Conclusion: The data from the microcosm study indicates that the risk to aquatic organisms from thiamethoxam potentially being emitted via STP to surface waters following the use as a wood preservative PT-8 at a treatment plant is acceptable.

2.2.4 Potential surface water contamination following leaching from noise barriers (in Service). The scenario assumes leaching from wood in service (noise barrier) to a moving water body. The values represented in the table below are the 30-day PECSW values.

Table 3: Thiamethoxam PECSW for following potential emissions from wood treatment in service (noise barrier)

Application Thiamethoxam Scenario PECSW (30 day) method application rate (μg ai/L) Brushing 0.1 g/m2 Noise barrier 0.0527 Vacuum pressure 15 g/m3 Noise barrier 0.134

No accumulation of thiamethoxam concentrations above these 30-day values is expected in these moving surface waters, since in addition to the expected dissipation/degradation in natural systems, the exchange/dilution in water is likely to result in daily total water exchange. The PECSW values for brushing and vacuum pressure treatments are 570 and 220 times lower than the microcosm NOEC and 1900 and 750 times lower than the microcosm NOEAEC, respectively. Another way to look at this is by assuming a concentration of 0.134 μg ai/L in the surface water (worst-case vacuum pressure scenario of noise barriers), but in addition also assume there is no

- 650 - Rl"VIS: Spain Thiamethoxam Doc III-A dissipation/degradation and dilution, but rather continued similar daily input of thiamethoxam to the surface water (i.e. highly unrealistic). It would take 220 days ofleaching of the 30-day leaching event for the PECsw to approach the microcosm NOEC. In reality leaching from treated wood will be weather event driven and peak concentrntions are expected to occur after rainfall events. Thus continuous exposure via leaching is not expected to occur. It should also be noted that the leaching model is based on worst-case laborato1y leaching data. Syngenta considers the results from the draft OECD studies to be ve1y conservative and do not represent the real life situation. Since thiamethoxam dissipated rapidly in the microcosm study. A comparison of the level of exposure to aquatic organisms over the first 21 -days of the microcosms study with a similar period relating to biocides emissions following leaching of wood in service is made. Previously the AUC for the measured concentrations in the NOEC and NOEAEC microcosm treatment groups was calculated (see 2.2.2). A similar approach was followed assuming constant exposure for 21 days to a concentrations of 0.0527 µg ai/L (brnshing treatment) and 0.134 µ.g ai/L (vacuum treatment) . Le unrealistic constant exposure to these 30-day PECsw. These are presented graphically below in Figure 4. This graph also compares these concentrations with those in the microcosm NOEC and NOEAEC treatments. Figure 4: Comparison between microcosm concentrations and PECSW resulting from thiamethoxam in OECD Noise arrier scenario

-+-Mesocosm 30 ug/L - Mesocosm 100 ug/L - PECsw brushing - PECsw vacuum

0 5 10 15 20 25 (Days)

The AUCs for brnshing and vacuum treatment were detennined to be 1.11 and 2.81 µg ai/L x d, respectively. These values indicate that the overall exposure to pelagic aquatic organisms in the microcosm study NOEC treatment group (AUC 100.49 µg ai/L x d) was approximately 90 and 36 times that following a 21 -day rather unrealistic worst-case exposure scenarios following brnshing and vacuum treatment of noise baITiers, respectively. In the case of the vacuum treated noise baniers, only after 750 days of constant exposure to this 30-day PECsw for thiamethoxam would the overall concentration to aquatic organisms in the surface waters be expected to exceed that of the microcosm study NOEC treatment. In the case ofbrnshed noise baniers this period would be longer still.

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In reality exposure in surface waters will be greatly reduced due to water exchange/dilution, dissipation and degradation in natural moving water systems. It is also reasonably expected that leaching events resulting in exposure to surface waters will be triggered by rainfall events and thus are expected to occur occasionally rather than continuously. Conclusion: The data from the microcosm study indicates that the risk to aquatic organisms from thiamethoxam potentially being entering surface waters via leaching from noise barriers in service following the use as a wood preservative PT-8 is acceptable.

3. CGA 322704 Litterbag study 2.1 Overall summary of litterbag study (Bader, 2001).

1 REFERENCE o Reference Bader, U. (2001): The effects of CGA 322704 (metabolite of thiamethoxam (CGA 293343)) on the decomposition of organic material in a field litterbag test. Syngenta Crop Protection AG, Basel, Switzerland, unpublished report No. 2002619.

The litterbag study is designed to evaluate the influence of treatment on the decomposition of organic material under field conditions. Soil micro-organsisms and macro-organsims are believed to have an important role in the decomposition of organic material and as such this study represents a functional endpoint for these organsisms. The metabolite of thiamethoxam (CGA 322704) was applied once at a rate equivalent to 70.7 g/ha directly to litterbags whilst on the surface of a cut meadow. Shortly after treatment, once the spray residues had dried, the litterbags were buried at approximately 5 cm depth among the roots of the meadow grass. Sampling of the bags on day 0, 33, 92, 155 and 275 revealed no relevant differences from the control in the degradation of organic material in the field following the application of CGA 322704. The No Observed Effect Rate for the study was therefore defined as 70.7 g/ha (the only rate tested). Since the litterbags were directly treated and then buried, assuming this application rate to the bags (equivalent to 70.7 g.ha) is homogenously distributed in a 10 cm soil layer of 1.5 g/cm3 density, around the bags the NOEC is 0.047 mg/kg soil. It should be noted that this was the only treatment group tested and so this NOEC is likely to be higher. Since the litterbag study is a field study it is considered that no additional safety factor be applied in defining the PNEC of 0.047 mg/kg soil from this study. PECsoil values for the metabolite CGA 322704 are presented in the table below.

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Table 4: PECsoil values for CGA 322704 following use of thiamethoxam as a wood preservative Scenario Compartment size (cm) PECsoil (mg/kg)a House 10 0.545 Brushing 100 g/m2 20 0.273 30 0.182 40 0.136 50 0.109 10 1.392 Vacuum 15 g/m3 20 0.696 30 0.464 40 0.348 50 0.278 Fence 10 0.436 Brushing 100 g/m2 20 0.218 30 0.145 40 0.109 50 0.0873 10 1.114 Vacuum 15 g/m3 20 0.557 30 0.371 40 0.278 50 0.223 Noise barrier 10 0.196 Brushing 100 g/m2 20 0.098 30 0.065 40 0.049 50 0.039 10 0.501 Vacuum 15 g/m3 20 0.251 30 0.167 40 0.125 50 0.100 a assuming 30% formation from parent thiamethoxam Assuming a 50 cm compartment size the PECsoil values marginally exceed the PNEC value from the litterbag study with the exception of the brushing treatment of noise barriers. However it should be noted that the PNEC is based on a NOEC that was the highest tested treatment in the litterbag study. In addition, the assumed 30% formation level of CGA 322704 from the peak thiamethoxam soil concentrations is conservative and the PECsoil values of thiamethoxam from which the metabolite is then formed are based on very conservative laboratory leaching data from studies conducted according to draft OECD guidance. Based on realistic leaching rates, the soil exposure to the metabolite CGA 322704 is expected to be significantly lower and as such within acceptable levels.

4. Overall Summary The thiamethoxam microcosm study indicates that the risk to aquatic organisms in surface waters potentially exposed to thiamethoxam entering surface waters via emissions from the treatment facility via a STP or leaching from noise barriers in service following the use as a wood preservative PT-8 is acceptable. Based on the litterbag study there is not expected to be any unacceptable effects on soil organisms following the use of thiamethoxam as a wood preservative PT-8. However, in view of the very conservative results from the leaching studies, we propose the following risk mitigation aspects: Further Application/label restrictions ( in line with Dichlofluanid and Tebuconazole on CIRCA)

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Losses during industrial/professional application by the dipping and automated enclosed spraying processes, as well as during tank cleaning, must be contained It is proposed that this risk is mitigated by restricting the storage of industrial treated timber to hard standing (preventing the direct losses to soil and to drains). For in situ treatment by brush (professional or amateur), wood preservative products must not be used to treat wooden structures located where direct losses to water cannot be prevented.

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