Annex to Regulation 283/2013 Diflufenican M-CA, Section

Sapec Agro S.A. / Globachem NV 1

Diflufenican

Herbicide

Application for Renewal of Approval according to

Commission Regulation 844/2012

DOCUMENT M-CA, Section 8

ECOTOXICOLOGICAL STUDIES ON THE

ACTIVE SUBSTANCE

Globachem NV
Brustem Industriepark
Lichtenberglaan 2019
3800 Sint-Truiden
Belgium

SAPEC AGRO SA
Avenido do Rio Tejo
Herdade das Praias
2910-440 Setúbal
Portugal

Author: SAPEC AGRO – 31.11.2016 Document ID: DFF-2016-11-CA-section 8-1.1

Annex to Regulation 283/2013 Diflufenican M-CA, Section
Sapec Agro S.A. / Globachem NV 2

Version history
Date Data points containing amendments or additions Document identifier and
and brief description version number

30.06.2016 Original version from applicant SAPEC Agro S.A. & DFF-2016-06-CA-section 8-
Globachem N.V. for submission to z-RMS United 1.0
Kingdom in the frame of AIR 3 re-submission

31.11.2016 Original version from applicant SAPEC Agro S.A. & DFF-2016-11-CA-section 8-
Globachem N.V. for submission to z-RMS United 1.1
Kingdom in the frame of AIR 3 re-submission

Author: SAPEC AGRO – 31.11.2016 Document ID: DFF-2016-11-CA-section 8-1.1

Annex to Regulation 283/2013 Diflufenican M-CA, Section
Sapec Agro S.A. / Globachem NV 3

Table of Contents

CA 8 ECOTOXICOLOGICAL STUDIES ON THE ACTIVE

SUBSTANCE ...........................................................................5
CA 8.1 Effects on Birds and Other Terrestrial Vertebrates ............................... 7
CA 8.1.1 Effect on birds .......................................................................................... 7
CA 8.1.1.1 Acute oral toxicity to birds ............................................................................... 7
CA 8.1.1.2 Short-term dietary toxicity to birds .................................................................. 8
CA 8.1.1.3 Sub-chronic and reproductive toxicity to birds .............................................. 8
CA 8.1.2 Effects on terrestrial vertebrates other than birds .............................. 10
CA 8.1.2.1 Acute oral toxicity to mammals...................................................................... 10
CA 8.1.2.2 Long-term and reproductive toxicity to mammals ........................................ 11
CA 8.1.3 Effects of active substance bioconcentration in prey of birds
and mammals ......................................................................................... 11
CA 8.1.4 Effects on terrestrial vertebrate wildlife (birds, mammals,
reptiles and amphibians) ....................................................................... 12
CA 8.1.5 Endocrine disrupting properties........................................................... 12
CA 8.2 Effects on Aquatic Organisms .............................................................. 14
CA 8.2.1 Acute toxicity to fish .............................................................................. 17
CA 8.2.2 Long-term and chronic toxicity to fish ................................................. 19
CA 8.2.2.1 Fish early life stage toxicity test .................................................................... 20
CA 8.2.2.2 Fish full life cycle test ..................................................................................... 20
CA 8.2.2.3 Bioconcentration in fish ................................................................................. 21
CA 8.2.3 Endocrine disrupting properties........................................................... 22
CA 8.2.4 Acute toxicity to aquatic invertebrates ................................................ 22
CA 8.2.4.1 Acute toxicity to Daphnia magna ................................................................... 22
CA 8.2.4.2 Acute toxicity to an additional aquatic invertebrate species ....................... 30
CA 8.2.5 Long-term and chronic toxicity to aquatic invertebrates ................... 30
CA 8.2.5.1 Reproductive and development toxicity to Daphnia magna ........................ 30
CA 8.2.5.2 Reproductive and development toxicity to an additional aquatic
invertebrate species........................................................................................ 31
CA 8.2.5.3 Development and emergence in Chironomus riparius ................................. 31
CA 8.2.5.4 Sediment dwelling organisms ........................................................................ 31
CA 8.2.6 Effects on algal growth .......................................................................... 35
CA 8.2.6.1 Effects on growth of green algae ................................................................... 35

Author: SAPEC AGRO – 31.11.2016 Document ID: DFF-2016-11-CA-section 8-1.1

Annex to Regulation 283/2013 Diflufenican M-CA, Section
Sapec Agro S.A. / Globachem NV 4

CA 8.2.6.2 Effects on growth of an additional algal species .......................................... 43

CA 8.2.7 Effects on aquatic macrophytes ........................................................... 45
CA 8.2.8 Further testing on aquatic organisms .................................................. 51
CA 8.3 Effects on Arthropods ........................................................................... 52
CA 8.3.1 Effects on bees....................................................................................... 52
CA 8.3.1.1 Acute toxicity to bees ..................................................................................... 52
CA 8.3.1.1.1 Acute oral toxicity .............................................................................................. 52
CA 8.3.1.1.2 Acute contact toxicity......................................................................................... 52
CA 8.3.1.2 Chronic toxicity to bees .................................................................................. 53
CA 8.3.1.3 Effects on honeybee development and other honeybee life stages ............ 56
CA 8.3.1.4 Sub-lethal effects ............................................................................................ 60
CA 8.3.2 Effects on non-target arthropods other than bees .............................. 60
CA 8.3.2.1 Effects on Aphidius rhopalosiphi .................................................................. 61
CA 8.3.2.2 Effects on Typhlodromus pyri ........................................................................ 61
CA 8.4 Effects on Non-Target Soil Meso- and Macrofauna ............................ 61
CA 8.4.1 Earthworms – sub-lethal effects ........................................................... 63
CA 8.4.2 Effects on non-target soil meso- and macrofauna (other than
earthworms)............................................................................................ 79
CA 8.4.2.1 Species level testing ....................................................................................... 79
CA 8.5 Effects on Nitrogen Transformation ..................................................... 96
CA 8.6 Effects on Terrestrial Non-Target Higher Plants ............................... 105
CA 8.6.1 Summary of screening data ................................................................ 106
CA 8.6.2 Testing on non-target plants ............................................................... 106
CA 8.7 Effects on Other Terrestrial Organisms (Flora and Fauna) .............. 109
CA 8.8 Effects on Biological Methods for Sewage Treatment ...................... 109
CA 8.9 Monitoring Data .................................................................................... 111

Author: SAPEC AGRO – 31.11.2016 Document ID: DFF-2016-11-CA-section 8-1.1

Annex to Regulation 283/2013 Diflufenican M-CA, Section
Sapec Agro S.A. / Globachem NV 19

Report date: 1996

Guidelines: OECD 203 (1992)
EEC 92/69 Method C1 (1992)
GLP: Yes
Previous evaluation: KIIA 8.2.1 in diflufenican DAR 2006
Material and methods: Results were relied upon for the first inclusion in Annex I to Directive
91/414/EEC.
Metabolite of diflufenican AE B107137 technical: purity 99%
A total of 70 fish (10 fish per concentration) were exposed to five
concentrations of diflufenican, a solvent control and a dilution water
control for a period of 96 hours. The definitive test was conducted at:
1.3, 2.5, 5.0, 10.0 and 20 mg/L. The nominal concentrations were
verified by chemical analysis at test initiation, after 72 hours of
exposure and test termination

Results
The 96-hour LC50 to rainbow trout after application of AE B107137 was calculated to be >17.3 mg/l based
on the mean measured concentrations. The NOEC was found to be 4.9 mg/l.

CA 8.2.2 Long-term and chronic toxicity to fish

In compliance with Reg 544/2011, it is not necessary to perform a chronic toxicity test on juvenile fish
when a fish early life stage toxicity test or a fish life cycle test has been performed while. In addition, no
fish juvenile growth test is recommended under Reg 283/2013. However, a study was reviewed during the
EU Review for diflufenican (SANCO/3782/08 – rev.1, 14 March 2008) and is provided as supportive
information and briefly summarized subsequently.
Ref. Point: KCA 8.2.2
Authors: Odin-Feurtet M., Lafay G., Soun A.
Title: Diflufenican: Fish, juvenile growth test (28 days) under flow-through
conditions
Company/Source: Bayer CropScience
Report No. R005623
Report date: 1997
Guidelines: OECD 215 (1994)
GLP: Yes
Previous evaluation: KIIA 8.2.2 in diflufenican DAR 2006
Material and methods: Results were relied upon for the first inclusion in Annex I to Directive
91/414/EEC.
Diflufenican technical
Nominal concentrations were exposed to a range of five concentrations
3, 8, 20, 40 and 80 μg/l under flow through conditions.

Author: SAPEC AGRO – 31.11.2016 Document ID: DFF-2016-11-CA-section 8-1.1

Annex to Regulation 283/2013 Diflufenican M-CA, Section
Sapec Agro S.A. / Globachem NV 20

Results
The 28 days flow through NOEC was found to be 19.2 μg/l after application of diflufenican technical to
Rainbow trout.

CA 8.2.2.1 Fish early life stage toxicity test

A fish early life-stage toxicity study was reviewed during the EU Review for diflufenican (SANCO/3782/08
– rev.1, 14 March 2008) and was considered to be acceptable. No additional studies have been conducted
since the review and none are considered necessary.
Ref. Point: KCA 8.2.2.1
Authors: Sousa J.
Title: Diflufenican – Early life-stage toxicity test with fathead minnow
(Pimephales promelas) under static conditions
Company/Source: Bayer CropScience
Report No. R0065752
Report date: 1998
Guidelines: FIFRA guideline 72.4
GLP: Yes
Previous evaluation: KIIA 8.2.2 in diflufenican DAR 2006 and in EFSA Scientific Report
(2007) 122
Material and methods: Results were relied upon for the first inclusion in Annex I to Directive
91/414/EEC.
Diflufenican technical: purity 96.8 %
The definitive test was conducted at: 0.62, 1.9, 5.6, 17 and 50 µg/L. The
nominal concentrations were verified by chemical analysis. Exposure
commenced within 24 h of egg fertilization and continued for 35 days,
to 30 days post hatch.

Results
The 35 days flow through NOEC was found to be 15 μg/l after application of diflufenican technical to
fathead minnow.

CA 8.2.2.2 Fish full life cycle test

Commission Regulation (EU) No. 283/2013 states that a fish full life cycle test may be required depending
upon the persistence and bioaccumulative potential of the active substance or if the substance is considered
as a potential endocrine disruptor.
The Panel recommends that FFLC-tests may be required where the BCF is > 1000, the elimination during
the 14 day depuration phase in the bioconcentration study is < 95 % or the substance is stable in water or
sediment (DegT90 > 100 days). Using the endpoints from the EFSA conclusion for Diflufenican: the BCF
for fish is 1596, elimination was > 97 % during the 14 day depuration phase in each tissue, diflufenican is

Author: SAPEC AGRO – 31.11.2016 Document ID: DFF-2016-11-CA-section 8-1.1

Annex to Regulation 283/2013 Diflufenican M-CA, Section
Sapec Agro S.A. / Globachem NV 21

stable to hydrolysis in water and the DT90 in sediment is >100 days. These criteria indicate that a full life-
cycle study may be required.
However, it is noted that the aquatic guidance is not definitive on this issue (only states a FFLC test may
be required), that the NOEC from the early-life stage study in the DAR (0.015 mg/L) is considerably higher
than the lowest endpoint for algae (ErC50 0.00045 mg/l) and therefore the risk to fish is not driving the
overall aquatic risk assessment.
Diflufenican is not suspected of being an endocrine disruptor, thus, on balance and in the interest of limiting
vertebrate testing it is not considered necessary to provide a full life-cycle study to avoid unnecessary
testing with aquatic vertebrates for animal welfare considerations as effects on reproduction of the parental
and the viability of the filial generation have been studied.

CA 8.2.2.3 Bioconcentration in fish

A fish bioconcentration study was reviewed during the EU Review for diflufenican (SANCO/3782/08 –
rev.1, 14 March 2008) since the log Pow for diflufenican is above than the trigger value of 3 and was
considered to be acceptable. No additional studies have been conducted since the review and none are
considered necessary.

Ref. Point: KCA 8.2.2.3

Authors: Briggs C.R., Chapleo S.
Title: (14C)-Diflufenican – Bioaccumulation and metabolism in rainbow trout
Company/Source: Bayer CropScience
Report No. R006596
Report date: 1998
Guidelines: OECD 305
GLP: Yes
Previous evaluation: KIIA 8.2.3 in diflufenican DAR 2006 and in EFSA Scientific Report
(2007) 122
Material and methods: Results were relied upon for the first inclusion in Annex I to Directive
91/414/EEC.
14
C-Diflufenican technical
Rainbow trout (Oncorhynchus mykis) were exposed to a flow-through
concentrations of 0.3 and 3.0 µg/L, for a 28 day period (uptake phase).
Fish were subsequently exposed to a continuous flow of dilution water
alone for a 14-day period. At the same time, a control experiment was
performed where fish were exposed to a continuous flow of dilution
water mixed with the same organic solvent as used to prepare the test.

Results
Bioconcentration factors were calculated for diflufenican, the principal radioactive component found in
both water and fish, based upon concentrations measured in fish tissues and water extracts. Mean
concentrations factors in whole fish for the steady-state period of 1276 and 1596 were found for the 0.3 and
3.0 µg as/L exposure levels, respectively. Depuration rate constants ranged from 0.289 to 0.298 day-1 at 0.3

Author: SAPEC AGRO – 31.11.2016 Document ID: DFF-2016-11-CA-section 8-1.1