Fludioxonil DAR 10 Vol3 B9 Part1 Public (Ecotoxicologia) Sinpassword

Draft Assessment Report (DAR)
- public version -

Initial risk assessment provided by the rapporteur Member State
Denmark for the existing active substance

FLUDIOXONIL

of the third stage (part A) of the review programme
referred to in Article 8(2) of Council Directive 91/414/EEC

Volume 3, Annex B, B.9, part 1

February 2006
Sun Aug 18 05:04:43 CEST 2013
European Commission
EU Review Programme
Fludioxonil
Volume 3
Annex B9: Ecotoxicology
Draft Assessment Report
Rapporteur Member State: Denmark
1anuary 2005, revised 1une 2005
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Fludioxonil Annex B9: Ecotoxicology January, revised June 2005
1
CONTENT
B.9 ECOTOXICOLOGY 3
Background 3
B.9.1 EFFECTS ON BIRDS (ANNEX IIA 8.1; ANNEX IIIA 10.1) 3
B.9.1.1 Acute oral toxicitv (ANNEX IIA 8.1.1, ANNEX IIIA 10.1.1) 3
B.9.1.2 Short-term dietarv toxicitv (ANNEX IIA 8.1.2) 6
B.9.1.3 Sub-chronic toxicitv and reproduction (ANNEX IIA 8.1.3) 10
B.9.1.4 Supervised cage or field trials (ANNEX IIIA 10.1.2) 16
B.9.1.5 Acceptance of bait, granules or treated seeds bv birds (ANNEX IIIA 10.1.3) 16
B.9.1.6 Effects of secondarv poisoning (ANNEX IIIA 10.1.4) 18
B.9.1.7 Summarv of effects on birds 19
B.9.1.8 Risk assessment for birds 21
B.9.2 EFFECTS ON AQUATIC ORGANISMS (ANNEX IIA 8.2; ANNEX IIIA 10.2) 22
B.9.2.1 Acute toxicitv to fish (ANNEX IIA 8.2.1, ANNEX IIIA 10.2.1) 22
B.9.2.2 Chronic toxicitv to fish (ANNEX IIA 8.2.2) 38
B.9.2.2.1 Chronic toxicity test on juvenile Iish (ANNEX IIA 8.2.2.1) 38
B.9.2.2.2 Fish early liIe stage toxicity test (ANNEX IIA 8.2.2.2) 51
B.9.2.2.3 Fish liIe cycle test (ANNEX IIA 8.2.2.3) 54
B.9.2.3 Bioconcentration in fish (ANNEX IIA 8.2.3) 54
B.9.2.4 Acute toxicitv to aquatic invertebrates (ANNEX IIA 8.2.4, ANNEX IIIA 10.2.1) 56
B.9.2.5 Chronic toxicitv to aquatic invertebrates (ANNEX IIA 8.2.5) 69
B.9.2.6 Effects on algae growth (ANNEX IIA 8.2.6, ANNEX IIIA 10.2.1) 77
B.9.2.7 Effects on sediment-dwelling organisms (ANNEX IIA, 8.2.7) 91
B.9.2.8 Effects on aquatic macrophvtes (ANNEX IIA, 8.2.8) 96
B.9.2.9 Microcosm or mesocosm studies (ANNEX IIIA 10.2.2) 98
B.9.2.12 Summarv of effects on aquatic organisms 104
B.9.2.12.1 Summary oI acute toxicity to Iish 104
B.9.2.12.2 Chronic toxicity to Iish 105
B.9.2.12.3 Bioconcentration in Iish 106
B.9.2.12.4 Acute and chronic toxicity to aquatic invertebrates 106
B.9.2.12.5 EIIects on algae growth 107
B.9.2.12.6 EIIect on sediment-dwelling organisms 108
B.9.2.12.7 EIIects on aquatic macrophytes 109
B.9.2.12.8 EIIects on aquatic micro/mesocosms 109
B.9.2.13 Risk assessment for aquatic organisms 110
B.9.3 EFFECTS ON OTHER TERRESTRIAL VERTEBRATES (ANNEX IIIA 10.3) 123
B.9.3.1 Summarv of effects on mammals 123
B.9.3.2 Risk assessment for mammals 124
B.9.4 EFFECTS ON BEES (ANNEX IIA 8.3.1; ANNEX IIIA 10.4) 125
B.9.4.1 Acute toxicitv (ANNEX IIA 8.3.1.1, ANNEX IIIA 10.4.1) 125
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2
B.9.4.2 Bee brood feeeding test (ANNEX IIA 8.3.1.2) 135
B.9.4.3 Residue test (ANNEX IIA 8.3.1.1, ANNEX IIIA 10.4.2) 135
B.9.4.4 Cage test (ANNEX IIIA 10.4.3) 135
B.9.4.5 Field tests (ANNEX IIIA 10.4.4) 136
B.9.4.6 Tunnel test (ANNEX IIIA 10.4.5) 136
B.9.4.7 Higher tier test (ANNEX IIA 10.4.3, 10.4.4, 10.4.4) 136
B.9.4.8 Summarv of effects on bees 136
B.9.4.9 Risk assessment for bees 137
B.9.5 EFFECTS ON OTHER ARTHROPOD SPECIES (ANNEX IIA 8.3.2; ANNEX IIIA 10.5) 138
B.9.5.1 Laboratorv, extended laboratorv, and semi-field tests (ANNEX IIA 8.3.2, ANNEX IIIA 10.5.1) 138
B.9.5.2 Field tests (ANNEX IIIA 10.5.2) 181
B.9.5.3 Summarv of effects on other arthropods 190
B.9.5.4 Risk assessment for other non-target arthropod species 195
B.9.6 EFFECTS ON EARTHWORMS (ANNEX IIA 8.4; ANNEX IIIA 10.6.1) 198
B.9.6.1 Acute toxicitv (ANNEX IIA 8.4.1, ANNEX IIIA 10.6.1.1) 198
B.9.6.2 Sublethal effects (ANNEX IIA 8.4.2, ANNEX IIIA 10.6.1.2) 207
B.9.6.3 Field studies (ANNEX IIIA 10.6.1.3) 215
B.9.6.4 Summarv of effects on earthworms 215
B.9.6.5 Risk assessment for earthworms 218
B.9.7 EFFECTS ON OTHER SOIL NON-TARGET MACRO-ORGANISMS 221
B.9.7.1 Sublethal - Effects on other soil non-target macro-organisms 221
B.9.7.2 Summarv of effects on other soil non-target macro-organisms 225
B.9.7.3 Risk assessment for other soil non-target macro-organisms 226
B.9.8 EFFECTS ON SOIL NON-TARGET MICRO-ORGANISMS (ANNEX IIA 8.5; ANNEX IIIA 10.7) 227
B.9.8.1 Impact on soil microbial activitv 227
B.9.8.2 Summarv of effects on soil non-target microorganisms 241
B.9.8.2.1 NitriIication 241
B.9.8.2.2 Respiration 242
B.9.8.3 Risk assessment for soil non-target micro-organisms 243
B.9.9 EFFECTS ON OTHER NON-TARGET ORGANISMS (FLORA AND FAUNA) BELIEVED TO BE AT RISK
(ANNEX IIA 8.6; ANNEX IIIA 10.8) 244
B.9.9.1 Fungicidal activitv (ANNEX IIA 10.6) 244
B.9.9.2 Summarv of fungicidal activitv 246
B.9.9.3 Terrestrial non-target plants (ANNEX IIIA 10.8) 247
B.9.9.4 Summarv of effects on Terrestrial non-target plants 252
B.9.9.5 Risk assessment for effects on Terrestrial non-target plants 252
B.9.10 EFFECTS ON BIOLOGICAL METHODS FOR SEWAGE TREATMENT (ANNEX IIA 8.7) 253
B.9.10.1 Summarv of impact of water treatment procedures 255
B.9.10.2 Risk assessment for sewage sludge treatment 255
B.9.11 REFERENCES RELIED ON 256
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4
Test substance: Fludioxonil (CGA 173506), Ciba-Geigy AG batch 805002 (stated purity 97.5).
Test species: Bobwhite quail, Colinus virginianus Irom D.R. & R.E. Wise, MonkIield, Bourn, Cambridgeshire,
England. The birds were young adults over sixteen weeks oI age.
Test concentrations: Control, 500, 1000, 2000 mg Iludioxonil/kg, not corrected Ior purity.
Test design: 40 adult Bobwhite quail were housed in groups oI 5 (males and Iemales separately) according to
treatment. Food was Standard HRC layer diet (batch 2212), and it was oIIered ad libitum. The duration oI the
study was as Iollows: Days 14 to 1 ~ 14 day pre-treatment period, day 0 ~ Day oI dosing and days 0 to 14 ~ 14
day post treatment period. Corn oil was used Ior the negative control and as a vehicle Ior the test compound.
Sampling: Mortality, health and clinical observations daily, Bodyweights weekly, Group mean Iood
consumption weekly. At termination oI the study post mortem examination was carried out on the ten birds
Irom the highest dose group.
Statistical analysis: None stated
Test conditions: stainless steel tiered cages with automatic drinker and Iood hopper, 7 hr. light 17 hr. dark,
humidity and temperature were recorded daily, Average minimum temperature was 15C with a standard
deviation oI 1.8C, average maximum temperature was 19C with a standard deviation oI 1.3C. The average
relative humidity was 75 with a standard deviation oI 6.2
Results:
Bodyweight changes and Iood consumption were variable and not diIIerent between treatment concentrations.
There were neither mortalities nor observations oI deviating health or clinical observations throughout the study
period.
At post mortem examination no abnormalities were observed.
LD
50
was higher than 2000 mg/kg (Table 9.1.1-1)
Table 9.1.1-1: Results Irom acute oral LD50 test on Bobwhite quail Colinus virginianus
Test substance
Test object
Fludioxonil (CGA 173506)
(Nominal mg /kg body weight)
Colinus virginianus
LD
50
NOEL (overall)
~2000
2000
Reviewers assessment:
The study was in accordance with the guideline and was acceptable. It was Iound that Iludioxonil (CGA 173506)
was not toxic to Bobwhite quail at the highest dose tested, i.e. 2000mg Iludioxonil/kg, thus LD
50
~ 2000 mg
as/kg.
REFERENCE: Annex KIIA 8.1.1
Report:
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7
Test species: Bobwhite quail, Colinus virginianus Irom D.R. & R.E. Wise, MonkIield, Bourn, Cambridgeshire,
England. The birds selected Ior the study was 8 days old and oI equal bodyweight.
Test concentrations: Three controls, and six nominal treatment concentrations, i.e. 163, 325, 650, 1300, 2600,
5200 ppm Iludioxonil (CGA 173506). Analysed concentrations were 162, 318, 655, 1290, 2600 and 4990 ppm
Iludioxonil (CGA 173506).
Test design: Nine groups, each oI ten birds, were allocated to treatment. The study period included a three day
pre-treatment period, a Iive day treatment period and a three day post treatment period.
Sampling: Mortality daily, Bird health and clinical observations daily, Group mean bodyweights days 3, 0, 5
and 8. Macroscopic post-mortem examination at termination Irom the highest surviving dose group.
Test conditions: Chicks were housed in wooden boxes (83x52x51 cm) with wire mesh lids, and they were Ied
Standard HRC chick diet (batch 4097). Food and water were available at libitum at all times. Humidity and
temperature were recorded daily. Average minimum temperature was 24C with a standard deviation oI 1.1C,
average maximum temperature was 27C with a standard deviation oI 0.9C. The average relative humidity was
41 with a standard deviation oI 4.0
Results:
No mortalities occurred. Blood was noted around the base oI the bills oI all birds in the group exposed to 325
ppm Iludioxonil. All other birds remained in good clinical health throughout the study.
Over days 5 to 8 a decrease in bodyweight was observed in the group exposed to 325 ppm Iludioxonil (Table
9.1.2-1).
Table 9.1.2-1: Average body weights oI Bobwhite quail at day 0, 5 and 8 in the acute dietary study with
Iludioxonil.
Treatment
(ppm as)
Average body weights (g) Food consumption (g)
0 hr. Test day 0 Test day 5 Test day 8 Test days 1-5 Test days 5-8
control 13 16.5 26.7 30.0 3.8 5.1
control 13.4 17.5 28.7 31.9 4.4 5.8
control 13.1 17.6 28.6 31.2 4.0 4.9
163 12.6 15.6 25.6 28.4 3.6 4.8
325 13.0 16.9 25.3 24.4 3.2 3.8
650 12.7 18.7 30.6 34.5 4.3 7.3
1300 13.5 16.5 26.1 29.3 3.1 4.5
2600 13.2 16.9 26.1 28.4 3.7 5.4
5200 13.1 17.0 24.1 27.3 3.3 5.1
Table 9.1.2-2: RMS calculation oI daily dose: Recalculation Irom mg as/kg to mg as/kg bw/day, based on the
general rule Ior the conversion, i.e. Daily dose (mg/kg bw/d) Concentration in Iood (mg/kg) multiplied by
daily Iood consumption (g per bird per day) divided by bodyweight (g).
Concentration in Iood
(ppm)
Average consumption
over 5-day exposure
period (g/day)
Average body over 5-day
exposure period (g)*
Daily dose
mg/kg bw/day
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(ppm)
Average consumption
over 5-day exposure
period (g/day)
Average body over 5-day
exposure period (g)*
Daily dose
mg/kg bw/day
5200 3.3 20.6 833
1300 3.1 21.3 189
650 4.3 24.7 113
Average oI day 0 and day 5
Table 9.1.2-3: Results Irom 8-day acute dietary toxicity study with Iludioxonil (CGA 173506).
Test substance Fludioxonil (CGA 173506)
Nominal (mg as/kg)
Fludioxonil
(mg as/kg bw/d)
Fludioxonil (CGA 173506)
Analysed (mg as/kg)
Test object Colinus virginianus
LC
50
(95 C.L.)-8-day ~5200 ~833 ~4990
LOEC 1300 189 1290
NOEC (sublethal) 650 113 655
The study Iollows the guideline and is acceptable. Since no mortalities were observed in any treatment group, the
LC
50
should be considered higher than 5200 ppm nominal (equivalent to ~ 833 mg as/kg bw/d). The observation
oI body weight decrease in the 325 ppm treatment group could not be explained. It is, however, not likely to be
associated with the treatment, since the pattern is not seen in any other treatment. Opposite to what is suggested
in the report, there is a small but signiIicant decrease in average body weight on test day 5 in the three highest
treatment concentrations when they are compared to the controls. This is also reIlected in the average Iood
consumption. ThereIore the LOEC and the NOEC should be 1300 ppm and 650 ppm Iludioxonil respectively
based on nominal concentrations.
Report:
Hakin, B., Rodgers, M., Anderson, A., & Dawe, S. (1993c). The dietary toxicity (LC
50
) oI CGA 173506 to the
Mallard duck. Syngenta Crop Protection Unpublished report CGA173506/0063. Huntingdon Research Centre
CBG 496/90384.
Guidelines:
EPA Pesticide assessment Guidelines, E, WildliIe and Aquatic Organisms, 71-2 Avian dietary LC
50
test.
Guideline deviations. None stated
GLP: Yes
Material and methods:
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9
The purpose oI the study was to determine the dietary toxicity (LC
50
) oI Iludioxonil to the Mallard duck.
Test substance: Fludioxonil (CGA 173506), Ciba-Geigy AG batch P805002 (purity 97.5).
Test species: Mallard duck, Anas platvrhvnchos Irom Mr. J. Coles, The County Game Farms, AshIord, Kent,
England. The birds selected Ior the study were 8 days old and oI equal bodyweight.
Test concentrations: Three controls, and six nominal treatment concentrations, i.e. 163, 325, 650, 1300, 2600,
5200 ppm Iludioxonil. Analysed concentrations were 165, 333, 671, 1310, 2610 and 5240 ppm Iludioxonil.
Test design: Nine groups, each oI ten birds, were allocated to treatment. The study period included a three day
pre-treatment period, a Iive day treatment period and a three day post treatment period.
Sampling: Mortalities daily, bird health and clinical observations daily, group mean bodyweights days 3, 0, 5
and 8. Macroscopic post-mortem examination at termination Irom the highest surviving dose group.
Test conditions: The Mallard ducklings were housed in galvanised steel, tiered brooders with wire mesh Iloors,
measuring 0.57x0.84x0.27 m. They were Ied Standard HRC chick diet (batch 4094). Food and water were
available at libitum throughout the study. Humidity and temperature were recorded daily, Average minimum
temperature was 27C with a standard deviation oI 3.3C, average maximum temperature was 31C with a
standard deviation oI 4.4C. The average relative humidity was 44 with a standard deviation oI 6.4
Results:
A total oI two mortalities occurred among birds exposed to 163 ppm Iludioxonil. No other mortalities occurred
and it is concluded that the LC
50
is higher than 5200 ppm Iludioxonil the maximum dose level applied.
Birds exposed to 2600 and 5200 ppm Iludioxonil produced comparatively darker excreta than birds exposed to
lower doses on days 4 and 5. All surviving birds remained in good health throughout the study. The macroscopic
post-mortem examination did not reveal any abnormalities.
All surviving control and test groups showed continued bodyweight increase throughout the study (Table 9.1.2-
4).
Table 9.1.2-4: Average body weights oI Mallard ducklings at day 0, 5 and 8 in the acute dietary study with
Iludioxonil (CGA 173506).
Treatment
(ppm as)
Average body weights (g) Food consumption (g)
0 hr. Test day 0 Test day 5 Test day 8 Test days 1-5 Test days 5-8
control 49 83 182 250 34 48
control 51 81 172 238 31 49
control 49 78 168 240 33 51
163 50 78 184 262 33 54
325 50 84 173 248 31 48
650 51 82 145 213 31 51
1300 49 84 173 248 33 52
2600 51 82 145 213 27 46
5200 49 83 144 225 26 47
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Sun Aug 18 05:04:43 CEST 2013
11
The purpose oI this study was to determine eIIects oI dietary administration oI Iludioxonil on reproduction in the
Bobwhite quail.
Test substance: Fludioxonil (CGA 173506), Batch No P91007 (purity 96.4 w/w)
Test species: Bobwhite quail, Colinus virginianus Irom BC Potter, Rosedean, Woodhurst, Cambridgeshire,
England. The birds were app. 9 months old at test start and approaching their Iirst breeding season.
Test concentrations: Control 0, 125, 300, 700 ppm Iludioxonil. Dose levels were selected on basis oI results
Irom a preliminary study.
Test design: Prior to the start oI the treatment period, birds were randomly allocated to cages with one male and
one Iemale in each cage. There were three test groups plus a control group and 20 cages (replicates) Ior each
treatment.
Sampling: Egg shell thickness, inIertile eggs, early and late embryonic death, Eggs were collected daily and aIter
the end oI the egg production period, intact eggs were incubated. Chicken weight, Iood consumption.
Statistical analysis: ANOVA (treatment, position), Williams test (comparisons oI treated groups with control)
Test conditions: The birds were housed in 5 batteries oI cages, each battery consisting oI 4 tiers oI 4 cages. Each
cage, which housed a replicate oI 1 male and 1 Iemale bird, was constructed oI polythene coated steel wire and
measured 0.31x0.39x0.24 m. The cages slooping Iloors with 0.1m egg catchers, and had externally attached Iood
hoppers and automatic drinkers. Birds were maintained under 7 hr. light : 17 hr. darkness until week 6. Then the
photoperiod was increased to 16 hr. and remained so until week 10. Then the photoperiod was increased by two
weekly halI hr. increments so that by week 11, 17 hr. light was provided. From week 17 until the end oI egg
production, the photoperiod remained at 17 hr. The mean light value was 94 lux, mean maximum and minimum
temperature were 23C and 21C respectively, and the mean relative humidity was 49. The basal diet was quail
layers diet without antibiotics and growth promoters. Test diets were Ied to the birds Ior 10 weeks during the
pre-egg laying period and Ior the 12 week egg production period. Controls received untreated basal diet.
Results:
The mean analysed concentration oI Iludioxonil (CGA 173506) varied 2.3 or less Irom the nominal
concentration as measured at week 1, 12 and 22 oI the study.
Dietary administration oI Iludioxonil (CGA 173506) at 125 ppm had no eIIect on adult birds oI Bobwhite quail
or their reproductive perIormance (Tables 9.1.3-1 to 6).
At 300 and 700 ppm a slight reduction in the proportion oI viable embryos oI eggs set was observed and
although not statistically signiIicant, the report concludes that it was probably oI biological importance (Table
9.1.3-2). At 700 ppm a signiIicant reduction in male bodyweight was observed at termination (Table 9.1.3-1).
Table 9.1.3-1: Adult group mean body weights, mean group consumption and egg production
Treatment
ppm
Sex Week oI study
Bodyweight (g) Consumption (g) Eggs laid
Cracked
Pre-egg laying Egg production
-2 0 22 1 10 22 Mean pr
Iemale
Total
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Sun Aug 18 05:04:43 CEST 2013
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Treatment
ppm
Sex Week oI study
Cracked
-2 0 22 1 10 22 Mean pr
Iemale
Total
0
M
F
190
190
193
188
205
235
13 19 23 19 62.6 1146 3.8
125
M
F
192
189
193
190
203
223
13 19 22 18 59.9 1154 2.3
300
M
F
190
187
192
189
201
231
13 18 23 18 55.4 981 5.0
700
M
F
188
187
191
191
195*
225
12 18 22 18 66.2 1316 3.3
*Statistically signiIicant (p0.05)
Table 9.1.3-2: Egg shell thickness, eggs set and survival oI embryos
Treatment
ppm
Egg shell
thickness
(mm)
Eggs
set
Viable
embryos
Live 3
week
embryos
viable embryos
oI eggs set
live 3-week embryos
oI viable embryos
Week
11
Mean Total Total Total Total Total
0 0.20 0.21 1020 927 915 91 99
125 0.20 0.20 1048 946 924 90 98
300 0.20 0.21 858 742 723 86 97
700 0.20 0.21 1180 1008 991 85 98
Table 9.1.3-3: Hatching results
Number oI normal
hatchlings
Number oI dead in shell normal
hatchlings oI
viable embryos
& normal hatchlings oI
live 3-week embryos
Treatment
ppm
Total Total Total Total
0 842 73 91 92
125 802 120 85 87
300 626 96 84 87
700 909 83 90 92
Table 9.1.3-4: Number oI 14 day survivors
Number oI 14-day
survivors
surviving oI eggs laid surviving oI
normal hatchlings
Number oI 14-day
survivors per Iemale
Treatment
ppm
0 747 65 89 41.0
125 705 61 88 36.6
300 555 57 89 31.5
700 810 62 89 40.8
Table 9.1.3-5: Group mean initial and 14-day bodyweights (g)
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Sun Aug 18 05:04:43 CEST 2013
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Initial bodyweight 14-day bodyweight
Treatment Mean Mean
0 6.9 25
125 6.7 25
300 6.7 25
700 6.7 25
Table 9.1.3-6: RMS calculation oI daily dose: Recalculation Irom mg as/kg to mg as/kg bw/day, based on the
general rule Ior the conversion, i.e. Daily dose (mg/kg bw/d) Concentration in Iood (mg/kg) multiplied by
daily Iood consumption (g per bird per day) divided by bodyweight (g).
(ppm)
Average consumption
over entire exposure
period (g/day)
Average body weight Ior
both sexes over exposure
period (g)
Daily dose
(mg/kg bw/day)
125 18 202.25 11.1
300 18 203.25 26.6
700 18 200.5 62.8
Table 9.1.3-7: Results Irom study oI eIIects oI dietary administration oI Iludioxonil (CGA 173506) on
reproduction in the Bobwhite quail.
Test compound Fludioxonil
mg as/kg bw/day
Fludioxonil
(mg as/kg)
Test object Bobwhite quail C. virginianus
LOEC (adult symptoms) 62.8 700 ppm
LOEC (reproduction ) 26.6 300 ppm
NOEC (adult symptoms)* 26.6 300 ppm
NOEC (reproduction ) 11.1 125 ppm
*based on decreased body weight
The study Iollows the guideline and is acceptable. According to the study report NOEC and LOEC Ior adult
symptoms were 300 and 700 ppm respectively based on decreased body weight, and NOEC and LOEC Ior
reproduction were reported to be 125 and 300 ppm, respectively. The RMS considers the NOEC Ior adult
symptoms, which is based on less than 5 reduction in body weight in males only, to be too conservative. Also
Ior reproduction the RMS considers the NOEC oI 125 ppm (equivalent to 11.1 mg as/kg bw/d) to be very
conservative and based on numerically very small diIIerences (5-6 compared to control) and with no dose
response relationship. As can be seen Irom table 9.1.3-4, the 'Number oI 14-day survivors per Iemale, which
can be considered an overall (integrated) measure oI reproductive success is not reduced at the highest test
concentration 700 ppm. The RMS thereIore Iinds that an overall NOAEC could be set to 700 ppm (equivalent to
62.8 mg as/kg bw/d).
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Sun Aug 18 05:04:43 CEST 2013
Sun Aug 18 05:04:43 CEST 2013
15
Table 9.1.3-8: Adult group mean body weights, mean group consumption and egg production
Treatment
ppm
Sex Week oI study
Cracked
-2 0 23 1 11 23 Mean pr
Iemale
Total
0
M
F
1179
1051
1047
935
1166
1128
169 190 266 211 39.4 1144 3.9
125
M
F
1179
1932
1063
887
1207
1103
137 180 226 194 34.6 929 2.5
300
M
F
1167
1035
1049
921
1164
1160
158 203 239 205 44.0 1314 2.4
700
M
F
1109
1043
1045
909
1140
1085
151 184 251 188 34.1 1026 3.1
Table 9.1.3-9: Egg shell thickness, eggs set and survival oI embryos
Egg shell
thickness
(mm)
Eggs
set
Viable
embryos
Live 3 week
embryos
viable embryos
oI eggs set
live 3-week
embryos oI viable
embryos
Treatment
ppm
Week
14
Mean Total Total Total Total Total
0 0.34 0.35 1021 826 779 81 94
125 0.34 0.34 845 732 674 87 92
300 0.35 0.35 1189 1149 1061 97 92
700 0.35 0.36 921 887 847 96 95
Table 9.1.3-10. Hatching results
Number oI
normal hatchlings
Number oI dead in
shell
normal hatchlings
oI viable embryos
& normal hatchlings oI
live 3-week embryos
Treatment
ppm
0 542 237 66 70
125 416 258 57 62
300 750 308 65 71
700 638 208 72 75
Table 9.1.3-11: Number oI 14-day survivors
Number oI 14-day
survivors
surviving oI eggs
laid
surviving oI
normal hatchlings
Number oI 14-day
survivors per Iemale
Treatment
ppm
0 516 45 95 17.8
125 405 44 97 15.0
300 734 56 98 24.5
700 627 61 98 21.0
Table 9.1.3-12: Group mean initial and 14-day bodyweights (g)
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Sun Aug 18 05:04:43 CEST 2013
17
Guidelines:
Based on elements Irom preliminary draIt proposal Ior OECD test guideline 'Tests Ior Avian Avoidance oI
Pesticide treated Seeds and Baits and BBA Guideline VI 25-1.
GLP: Yes
The purpose oI the study was to evaluate whether Rock dove Columba livia avoids treated wheat seed Triticum
spp.
Test substance: Untreated Wheat ST 00 TA OCH and Treated Wheat ST 22 TA 575, wheat treated with A 8207 I
(CELEST 025 FS). A 8207 I was applied at 49.4 mg Iludioxonil/kg seed.
Test species: Rock dove Columba livia Irom Conquest Farm, Centreville, MD 21617. The doves ranged in
weight Irom 434 to 685 g the day prior to test initiation. All birds were Irom the same hatching season, pen
reared and phenotypically indistinguishable Irom wild birds. All test birds were preconditioned to test conditions
Ior 5 weeks prior to the test.
Test concentrations: A 8207 I was applied at 49.4 mg Iludioxonil/kg seed.
Test design: The treatment group and the control group consisted oI six replicates oI 2 doves each. Rock doves
in the treatment group were presented a diet consisting oI treated seed Ior a six hr. period beginning in the
morning oI each day on three successive days. The birds were then given untreated Iood Ior a two hr. period.
Rock doves in the control group were presented untreated wheat seed Ior both the six hr. and two hr. Ieeding
periods during the same three days. Acclimation to untreated wheat and Ieeding trays Ior 6 days, 15-hr.
overnight prior to a 6-hr. exposure period on each oI three consecutive days, Iollowed by 2-hr. presentation oI
untreated seed. Post exposure observation lasted 6-days.
Sampling: Food consumption, sublethal clinical signs and mortality.
Statistical analysis: One-tailed Student`s T-test.
Test conditions: The birds were housed indoor in batteries oI pens, each pen contained two birds. The birds were
maintained at room temperature. Average temperature was 23.6C with an average relative humidity oI 77. A
photoperiod oI 9 hr. oI light and 17 hr. oI darkness was controlled with an automatic timer. The test area was
illuminated with Iluorescent lights at an intensity oI 103 to 327 Lux.
Results:
There were no mortalities in any group during the course oI the study. All birds in both groups were normal in
appearance and behaviour throughout the test.
Mean bodyweights were comparable between the control and treatment group during the course oI the study
(Table 9.1.5-1).
During the six hr. exposure phase there was an average decrease oI 23, i.e. Irom 12 33 in the mean
consumption oI birds in the treatment group relative to the birds in the control group (Table 9.1.5-2).
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Table 9.1.5-1: Body weights Irom avoidance test with rock dove
Day -6 Day -1 Day 3 Day 8 Total change change
Control Mean 533 531 526 546 15 3
SD 42 46 45 38 17 3
49.4 mg
as/kg seed
Mean 539 535 555 547 12 2
SD 66 69 73 70 11 2
Table 9.1.5-2: Feed consumption (g/bird/day) Irom avoidance test with rock dove during the exposure phase.
Day 0 Day 1 Day 2 Mean
6 hr 2 hr 6 hr 2 hr 6 hr 2 hr 6 hr 2 hr
Control Mean 20.2 11.8 16.6 10.7 18.0 9.6 18.3 10.7
SD 5.6 3.2 5.2 3.9 5.1 43.8 5.0 3.6
49.4 mg
as/kg seed
Mean 15.8 14.7 14.7 15.0* 12.0* 15.7* 14.1 15.1
SD 3.2 3.9 4.1 3.9 2.9 3.2 3.0 3.1
oI
control
78 124 88 141 67 163 77 141
*Statistically signiIicant Irom the control (p0.05)
The authors state no guideline deviations however the OECD draIt proposes a 'no choice test whereas this
study was conducted as a 'choice in time test (i.e. only a 6 h exposure period with no choice). The notiIier
argues that this would resemble a natural exposure situation. The RMS Iinds this questionable as a longer time
period, Ior instance 24 hr, would seem more realistic. The results are presented as 23 reduction in the
consumption oI the treated seed as compared to the non-treated, which would be equivalent to an avoidance
Iactor oI 0.77 Ior rock doves, Columba livia. However, RMS questions the generality oI these Iindings and the
relevance Ior other bird species. The study in principle is acceptable, but does not represent a realistic exposure
situation and has thereIore not been used in the risk assessment, nor been relied upon.
B.9.1.6 EFFECTS OF SECONDARY POISONING (ANNEX IIIA 10.1.4)
The assessment oI eIIects oI secondary poisoning is based on the study presented in Table 9.2.3-3 section B.
9.2.3. 'Bioconcentration Iactors in Iish (juvenile bluegill sunIish, Lepomis macrochirus) based on
14
C-labelled
Iludioxonil (CGA 173506). The calculations presented in Table 9.1.6-1 and 9.1.6-2 was carried out as Iollows:
As a worst case the FOCUS step 1 PEC
max
Ior water was used Ior the chronic assessment and multiplied with the
BCF (bio-concentration Iactor) to obtain PEC
Iish
. Then PEC
Iish
was multiplied by a 'Bird Factor oI 0.21 to
convert PEC
Iish
to daily dose Ior birds. The bird Iactor value oI 0.21 is based on a 1000 g bird eating 206 g Iish
per day. The background Ior the calculations are described in SANCO/4145/2000 Iinal oI 25 September 2002
'Guidance Document on Risk Assessment Ior Birds and Mammals under Council Directive 91/414/EEC.
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Table 9.1.6-1: Calculation oI chronic toxicity exposure ratios to Iish consuming birds, based on bioconcentration
oI Iludioxonil in Iish.
PEC
max
water
BCF PEC
Iish
Bird
Iactor
Daily
dose
NOAEL
bird
TER bird Trigger
Whole-body
0.0142 366 5.197 0.21 1.091 62.8 57 5
Edible parts
0.0142 58 0.824 0.21 0.173 62.8 360 5
Non-edible
parts
0.0142 741 10.52 0.21 2.210 62.8 28 5
It is evident that the TER values exceed the trigger values Ior the chronic assessment, thereby no risks oI
secondary poisoning oI Iish eating birds are indicated.
No basis Ior calculations oI secondary poisoning Irom earthworms (BCF-study) to earthworm eating birds was
presented in the documentation submitted by the notiIier. However using the approach Irom the 'Guidance
Document on Risk Assessment Ior Birds and Mammals under Council Directive 91/414/EEC a BCF can be
calculated as:
BCF (0.84 0.01 K
ow
) / I
oc
K
oc
(0.84 0.01 x 10
4.12
) /0.02 x 12000 0.46
K
ow
Partitioning coeIIicient, log Pow 4.12 Ior Iludioxonil
K
oc
Organic carbon adsorption coeIIicient 12000 (lowest value worst case)
I
oc
Organic carbon content oI soil 0.02 (deIault value)
Since BCF is 1 no risks oI secondary poisoning oI earthworm eating birds is indicated.
B.9.1.7 SUMMARY OF EFFECTS ON BIRDS
A total oI seven bird studies were submitted. Two oI these investigated the acute oral toxicity oI Iludioxonil
(CGA 173506) to Bobwhite quail Colinus virginianus and Mallard duck Anas platvrhvnchos. The conclusion
Irom these studies was that NOEL Ior acute oral mortality oI Iludioxonil to birds was 2000 mg /kg bw, i.e. the
highest concentration tested. Regurgitation occurred at the lowest dose tested, i.e. 51.2 mg as/kg body weight,
and all higher doses in the study with Mallard duck. In the study with the Bobwhite there were no signs oI
toxicity and the LD
50
was higher than 2000 mg Iludioxonil/kg bw. Concerning the regurgitation in the Mallard
duck study the Guidance Document on Terrestrial Ecotoxicology Under Council Directive 91/414/EEC (rev 2,
October 2002) states that iI regurgitation is recorded in all doses, a study with a species that does not regurgitate
should be required. In this case a valid study on Bobwhite quail is available, and no Iurther data are needed.
Another two studies investigated the short-term dietary toxicity oI Iludioxonil (CGA 173506) to the same two
bird species. For both species LC
50
was higher than 5200 ppm (equivalent to ~ 833 mg as/kg bw/d Ior Bobwhite
quail), i.e. the highest dose tested, NOEC was 650 ppm Ior Bobwhite quail and 1300 ppm Ior Mallard duck. No
mortalities oI birds in any oI the submitted studies could be related to the test substance.
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Two studies concerning reproductive and subchronic toxicity were submitted. In the study with Bobwhite quail
NOEC and LOEC Ior reproduction were reported to be 125 (equivalent to 11.1 mg as/kg bw/d) and 300 ppm,
respectively. The RMS, however, considers the NOEC Ior reproduction to be very conservative and based on
numerically very small diIIerences (5-6 compared to control) and with no dose response relationship. Since the
'Number oI 14-day survivors per Iemale, which can be considered an overall (integrated) measure oI
reproductive success is not reduced at the highest test concentration 700 ppm the RMS Iinds that an NOAEC
could be set to 700 ppm (equivalent to 62.8 mg as/kg bw/d). For Mallard duck the NOEC was 700 ppm, i.e. the
highest concentration tested.
Additionally an avoidance test was submitted Ior Rock dove, Columba livia exposed to wheat seed treated on the
surIace with CELEST 025 FS (A 8207 I) containing 49.4 mg as per kg seed. This study resulted in 23 lower
average consumption oI treated seeds than oI control seeds during the 6 h exposure. Consequently the avoidance
Iactor could be 1 0.23 0.77 Ior rock doves. However, RMS questions the generality oI these Iindings and the
relevance Ior other bird species. The result will not be used in the risk assessment.
Table 9.1.7-1: Summary oI eIIects on birds.
ReIerence Species Test substance Type NOEL/NOEC LD
50
/LC
50
IIA 8.1.1
Hakin et al.
(1993a)
Bobwhite
quail
Fludioxonil
(CGA 173506)
97.5 purity
Acute oral 2000 mg as/kg bw ~2000 mg as/kg
bw
IIA 8.1.1
Hakin et al.
(1990)
Mallard
duck
Fludioxonil
(CGA 173506)
97.5 purity
Acute oral
(Vomiting)
2000 mg as/kg bw
( 51.2 mg as/kg)
~2000 mg as/kg
bw
IIA 8.1.2
Hakin et al.
(1993b)
Bobwhite
quail
Fludioxonil
(CGA 173506)
97.5 purity
Short term dietary 650 mg as/kg ~5200 mg as/kg
(~ 833 mg/kg
bw/d)
IIA 8.1.2
Hakin et al.
(1993c)
Mallard
duck
Fludioxonil
(CGA 173506)
97.5 purity
Short term dietary 1300 mg as/kg ~5200 mg as/kg
(~ 1191 mg/kg
bw/d)
IIA 8.1.3
Johnson et
al. (1996a)
Bobwhite
quail
Fludioxonil
(CGA 173506)
96.4 purity
Subchronic and
reproductive
toxicity
125mg as/kg Report
(11.1 mg/kg bw/d)
700 mg as/kg RMS
(62.8 mg/kg bw/d)
IIA 8.1.3
Johnson et
al. (1996b)
Mallard
duck
Fludioxonil
(CGA 173506)
96.4 purity
Subchronic and
reproductive
toxicity
700 mg as/kg
(126 mg/kg bw/d)
IIIA2 10.1.3
Gallagher,
S.P. &
Beavers, J.B.
1999
Rock dove CELEST 025
FS (A 8207 I)
49.4 mg/kg
wheat seed
Food avoidance
6. h exposure
23 Iood
avoidance~
A 1-0.23 0.77*
* The generality and relevance Ior other bird species is questionable.
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21
B.9.1.8 RISK ASSESSMENT FOR BIRDS
Toxicity Exposure Ratio (TER) and Estimated Theoretical Exposure (ETE) were calculated based on the
methods, i.e. equations and tabulated values described in the 'Guidance Document on Risk Assessment Ior Birds
and Mammals Under Council Directive 91/414/EEC (Sanco/4145/2000 oI 25. September 2002).
Residues per Unit Dose values Ior short grass and small insects were used as suggested in the guidance
document Ior seed treatment and orchard/vine. As standard indicator species a granivorous bird and an
insectivorous bird were used. The most important constants and variables Ior the TER calculations and the
resulting acute and short- and long-term TER values Ior birds are presented in Table 9.1.8-1. 'Avoidance
Iactor, 'Fraction oI diet obtained Irom area and 'Fraction oI Iood type in diet were all assumed to be 1.
For the short-term and sub-chronic studies the endpoints were recalculated to a daily dose based on the body
weight and mean Iood consumption obtained in the studies. For calculations see Tables 9.1.2-2, 9.1.2-5, 9.1.3-6
and 9.1.3-13.
The resulting TER values are presented in Table 9.1.8-1. The scenarios Ior seed treatment/granivorous bird result
in TER values oI 86857, 36176, and 482 respectively Ior acute, short-term, and long-term/sub-chronic toxicity
(Table 9.1.8-1). For vine/insectivorous birds, TER-values Ior acute and short- term exposures were respectively
160 and 119. These TERs are higher than the trigger value oI 10 Ior acute and short-term toxicity. For
reproductive toxicity the TER is 1.6 based on the NOEC oI 11.1 mg/kg bw/day (125 ppm) which was based on
very small (5-6) reproductive eIIects. However iI based on the more ecologically relevant NOAEL oI 62.8
mg/kg bw/d (700 ppm) the TER is 9 and thus higher than the trigger value oI 5, and thereIore acceptable. Based
on these considerations both the scenarios Ior seed treatment and Ior vine produce TERs higher than the trigger
values Ior acute toxicity, short term and subchronic toxicity. The RMS thereIore considers that the intended use
oI Iludioxonil does not present an unacceptable risk to birds.
Calculations oI daily dose TERs Ior Iish consuming birds Irom the Iish bioconcentration study did not indicate
any unacceptable risks (see B.9.1.6, Tables 9.1.6-1 and 9.1.6-2).
Table 9.1.8-1: Calculated TER values Ior acute, short-term, sub-chronic risk Ior birds Ior an application rate oI
0.250 kg as/ha.
Type Bird type Scenario FIR/bw RUD AR C
(RUD x
AR
ETE Toxicity
Daily
dose
TER
Acute Granivorous Seed
treatment
0.38 1 0.00875 0.00875 0.0230 2000 86857
Acute Insectivorous Vine 1.04 52 0.25 13 12.5 2000 160
Acute Whole Iish Vine See Table 9.1.6-1 2000 1832
Short-term Granivorous Seed
treatment
0.38 1 0.00875 0.00875 0.0230 833 36176
Short-term Insectivorous Vine 1.04 29 0.25 7.25 6.97 833 119
Subchronic Granivorous Seed
treatment
0.38 1 0.00875 0.00875 0.0230 11.1 482
Subchronic Insectivorous Vine 1.04 29 0.25 7.25 6.97 11.1* 1.6*
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23
Test conditions: Flow-through. The rainbow trout were held in water Irom the same source as that used during
the test. AIter hatching, the Iishes were held 69 days prior to the test. The Iishes were acclimated to test
conditions Ior 70 hr. prior to test initiation. Flow-through in 25-L polyethylene aquaria Iilled with 15-L oI test
water. The photoperiod was 16 hr. oI light (430 lux) and 8 hr. oI darkness. Temperature was between 11.3 and
11.9C during the test. The pH was between 8.0 and 8.5 during the test.
Results:
Nominal test concentrations oI Iludioxonil were 0.19, 0.32, 0.54, 0.90, 1.5 mg/L, whereas measured test
concentrations oI Iludioxonil were 0.17, 0.26, 0.38, 0.71, 0.69 mg/L. The measured concentrations were used Ior
the assessment oI the toxicity oI Iludioxonil to rainbow trout in Table 9.2.1-1 and 9.2.1-2.
Daily observations oI mortality and other signs oI toxicity are presented in Table 9.2.1-1. Rainbow trout in the
control chambers appeared healthy and normal throughout the study period. Results on LC
50
, LOEC and NOEC
are presented in Table 9.2.1-2.
Table 9.2.1-1: Cumulative percent mortality (M) and number oI Iish with treatment related eIIects (E)* Irom
acute toxicity study with Oncorhvnchus mvkiss.
2 hr. 24 hr. 48 hr. 72 hr. 96 hr. Mean measured
concentration (mg
as/L)
Replicate
M E* M E* M E* M E* M E*
Negative control A
B
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Solvent control A
B
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0.17 A
B
0
0
0
0
0
0
10
10
0
0
10
10
0
0
10
10
0
0
10
10
0.26 A
B
0
0
2
1
0
0
10
10
0
0
10
10
0
0
10
10
0
0
10
10
0.38 A
B
0
0
10
10
0
0
10
10
0
0
10
10
0
10
10
9
0
30
10
7
0.71 A
B
0
0
10
10
100
100
-
-
100
100
-
-
100
100
-
-
100
100
-
-
0.69 A
B
0
0
10
10
60
70
4
3
100
100
-
-
100
100
-
-
100
100
-
-
* Treatment related eIIects included lethargy, discoloration and loss oI equilibrium
Table 9.2.1-2: Results Irom 96-hr. acute toxicity study with Iludioxonil on Oncorhvnchus mvkiss under Ilow
through conditions
Test substance Fludioxonil (mg as/L)*
Test object Rainbow trout Oncorhynchus mykiss
LC
50
(95 C.L.)-96-hr. 0.47 (0.38 0.69)
LOEC 0.17
NOEC (Mortality) 0.26
NOEC (Sublethal) 0
*Based on measured concentrations
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25
dissolved oxygen ranged Irom 7.7 to 9.4 mg/L, and pH Irom 6.7 to 7.1. The maximum loading oI Iish during the
exposure period was 0.084 g oI biomass per liter oI test solution per day.
Results:
Analyses resulted in measured concentrations, which averaged 90 oI nominal. Throughout the pretest and
deIinitive exposure, no visible sign oI precipitate or undissolved material was present in the test vessels or diluter
apparatus.
Table 9.2.1-3: Cumulative percent mortality oI rainbow trout Oncorhvnchus mvkiss exposed to Iludioxonil
(CGA 173506) technical during a 96-hr. Ilow-through acute toxicity test.
Mean measured
concentration
Cumulative mortality ()
(mg as/L) 24-hr. 48-hr. 72-hr. 96-hr.
Solvent control 0 0 0 0
Control 0 0 0 0
0.066 0 0 0 0
0.11 0
cd
0
b
0
b
0
a
0.18 0
b
0
ce
10
ce
10
a
0.33 20
a
50
a
100 100
0.54 100 100 100 100
a
All surviving Iish exhibited a complete loss oI equilibrium
b
All surviving Iish exhibited a partial loss oI equilibrium
c
Several surviving Iish exhibited a partial loss oI equilibrium
d
Several surviving Iish exhibited lethargic behavior
e
Several surviving Iish exhibited a complete loss oI equilibrium.
Table 9.2.1-4: Results Irom acute toxicity study with Iludioxonil (CGA 173506) technical on Oncorhvnchus
mvkiss in a 96-hr. Ilow-through test.
Test object Oncorhvnchus mvkiss
LC
50
(95 C.L.)-96-hr. 0.23 (0.18 0.33)
LOEC 0.11
NOEC (Sublethal) 0.066
The study was acceptable, and in accordance with the guideline. The LC
50
was 0.23 mg as/L, sublethal eIIects
were observed at concentrations higher than 0.066 mg as/L, i.e. the NOEC. The lowest concentration at which
sublethal eIIects were observed was 0.11 mg as/L, and the lowest concentration at which lethal eIIects were
observed was 0.18 mg as/L, where the mortality was 10 (Table 9.2.1-3 and table 9.1.2-4).
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27
0.74 mg as/L Ior bluegill sunIish exposed to Iludioxonil (CGA 173506). NOEC Ior sublethal eIIects was 0.14
mg as/L. NOEC Ior mortality was 0.45 mg as/L, and LOEC was 0.26 mg as/L (Table 9.2.1.5 and Table 9.2.1.6).
Table 9.2.1-5: Cumulative percent mortality oI bluegill sunIish Lepomis macrochirus exposed to Iludioxonil
(CGA 173506) technical during a 96-hr. Ilow-through acute toxicity test.
Mean measured Iludioxonil
concentration
Cumulative mortality ()
(mg as/L) 24-hr. 48-hr. 72-hr. 96-hr.
Solvent control 0 0 0 0
Control 0 0 0 0
0.14 0 0 0 0
0.26 0
d
0
d
0
d
0
d
0.45 0
bc
0
a
0
a
0
a
0.62 0
a
0
a
0
a
10
a
0.96 100 100 100 100
a All surviving Iish exhibited a complete loss oI equilibrium
b Several surviving Iish exhibited a complete loss oI equilibrium.
c Several surviving Iish exhibited a partial loss oI equilibrium
d Several surviving Iish exhibited a partial loss oI equilibrium
'
c
&
d
both denotes 'Several surviving Iish exhibited a partial loss oI equilibrium. This is probably due to a typing error in
the original report
Table 9.2.1-6: Results Irom acute toxicity study with Iludioxonil (CGA 173506) technical on bluegill sunIish
Lepomis macrochirus in a 96-hr. Ilow-through test.
Test substance Fludioxonil(mg as/L)*
Test object Lepomis macrochirus
LC
50
(95 C.L.)-96-hr. 0.74 (0.62 0.96)
LOEC 0.26
The study is accepted and in agreement with the guideline. The conclusion oI this 96-hr. acute toxicity study
under Ilow-through conditions was that the LC
50
was 0.74 mg as/L Ior bluegill sunIish exposed to Iludioxonil
(CGA 173506). NOEC Ior sublethal eIIects was 0.14 mg as/L. NOEC Ior mortality was 0.45 mg as/L, and
LOEC was 0.26 mg as/L (Table 9.2.1.5 and Table 9.2.1.6).
Report:
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30
US EPA FIFRA-Guideline No. 72-1
Guideline deviations. The Iollowing deviations to the US EPA FIFRA-Guideline No. 72-1 were mentioned:
Number oI replicates, oI test organisms, continuous monitoring oI temperature, and equilibration time.
GLP: Yes
The purpose oI the study was to evaluate the 96-hr. acute toxicity oI CGA 339833 (metabolite oI Iludioxonil) to
rainbow trout (Oncorhvnchus mvkiss) under static conditions.
Test substance: CGA 339833 (metabolite oI Iludioxonil, CGA 173506), Batch No. KI 6119/6, purity 90.
Test species: Rainbow trout Oncorhvnchus mvkiss Irom P. Hohler, Zeiningen, Switzerland. Mean length oI Iish
was 57mm (56 59mm), Mean weight oI Iish was 1.54g ( 1.40 1.70g).
Test concentrations: Based on the results Irom the range Iinding test, a limit test was perIormed with a nominal
concentration oI 100 mg CGA 339833/L, and a negative control.
Test design: 7 Iish per concentration and control, 7 Iish per aquarium.
Sampling: Mortality, abnormal behaviour and signs oI intoxication.
Statistical analysis: Not possible, since no eIIects were observed
Test conditions: Static conditions in 15 L oI water in 20 L aquaria, Loading was 0.72 g Iish/L, Temperature 13.3
13.4C, pH was 7.9 to 8.1, dissolved oxygen was 89 to 100 oI saturation. Fluorescent light 16 hr. daily with
30 minutes transition period.
Results:
The concentration oI the test substance was homogeneously distributed in the test vessel, no precipitation was
observed. In the control and the treatment concentration (100 mg/L) no mortalities, no signs oI intoxication, and
no signs oI abnormal behaviour was observed during the test.
Table 9.2.1-9: Results Irom 96-hr. acute toxicity study under static conditions with CGA 339833 (metabolite oI
Iludioxonil) on rainbow trout Oncorhvnchus mvkiss.
Test substance Fludioxonil metabolite
CGA 339833 (mg as/L)
LC
50
(95 C.L.)-96-hr. ~100
LOEC ~100
NOEC (Mortality) 100
NOEC (Sublethal) 100
Principally a study should use the newest available version oI the guidelines applied. Consequently a study dated
year 2000, should use the US-EPA guideline OPPTS 850.1075 Fish Acute Toxicity Test Irom 1996, not the
older US-EPA FIFRA guideline 72-1. However, the deviations to the US-EPA FIFRA guideline 72-1, have no
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33
Guidelines:
OECD Guideline NO. 203, Fish Acute Toxicity Test, dynamic system.
Guideline deviations. Mean length oI Iish exceeded guideline recommendations, i.e. Iish were 62 mm as
opposed to the recommendation oI 50 +10mm.
GLP: Yes
The purpose oI the study was to evaluate the 96-hr. acute toxicity oI SWITCH 62.5 WG to Rainbow trout under
dynamic conditions.
Test substance: SWITCH 62.5 WG (A 9219 B), Batch No. P.311001, purity not applicable, Composition
Iludioxonil 25.1 and cyprodinil 37.6.
Test species: Rainbow trout Oncorhvnchus mvkiss Irom P Hohler/Ch-4314 Zeiningen. Length: mean 62 mm (57
66 mm), Weight: mean 2.01 g (1.61 2.47 g), Loading 0.07 g/L in 24 hr.
Test concentrations: Nominal - control, 0.23, 0.50, 1.1, 2.3, 5.0 mg/L, actual control, 0.20, 0.52, 1.3, 3.0, 7.0
mg cyprodinil/L.
Test design: 7 Iish per concentration, control and aquarium.
Sampling: Mortalities and sublethal signs oI toxicity as well as oxygen content, pH and temperature aIter 0, 2-4,
24, 48, 72 and 96 hr. oI exposure.
Statistical analysis: The LC-50 values were calculated according to Berkson, JASA 48 (1953, 569 599. LC-
values were also graphically determined on gausso-logarithmic probability paper.
Test conditions: Dynamic conditions, T 15C, pH 7.8 8.2, O
2
82 97. 20 L glass aquaria Iilled with 15
L (36x22x25cm
3
), 16 hr. light, 8 hr. dark.
Results:
LC
50
was 2.1 mg Iludioxonil
1
/L aIter 72 and 96 hr. (Table 9.2.1-12), aIter 24 and 48 hr. LC
50
was 3.5 and 3.1 mg
Iludioxonil
1
/L, respectively
acute toxicity study with SWITCH 62.5 WG (A 9219 B ) on Oncorhvnchus mvkiss under dynamic conditions.
Calculated
concentration
(mg
Iludioxonil
/L)
Concentration
actual mean
(mg
cyprodinil/L)
2-4 hr. 24 hr. 48 hr. 72 hr. 96 hr.
Control Control 0 0 0 0 0 0 0 0 0 0
0.13 0.20 0 0 0 0 0 0 0 0 0 0
0.35 0.52 0 0 0 x 0 x 0 x 0 x

1
Fludioxonil concentrations were calculated Irom the cyprodinil:Iludioxonil ratio in the Iormulated product as
given in the material and methods section.
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Sun Aug 18 05:04:43 CEST 2013
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35
Mean length oI Iish: 44 mm (40 50mm) since mortality is too high with smaller sized Iish.
Oxygen at 58 oI saturation value aIter 48h exposure at concentration 0.6 and 1.3 mg/L Ior a Iew hr. (p13
chapter 4.5).
GLP: Yes
The purpose oI the study was to evaluate the 96-hr. acute toxicity oI SWITCH 62.5 WG

to Bluegill sunIish.
Test substance: SWITCH 62.5 WG (A 9219 B ), Batch No. P.311001, purity not applicable, Composition
Iludioxonil 25.1 and cyprodinil 37.6.
Test species: Bluegill sunIish Lepomis macrochirus Irom Ozage CatIish. Box 222 Missouri 65065/USA. Length:
mean 44 mm (40 50 mm), Weight: mean 0.85 g (0.52 1.46 g), Loading 0.40 g/L.
Test concentrations: Nominal - control, 0.4, 0.87, 1.9, 4.1, 9.0 mg/L, actual control, 0.2, 0.6, 1.3, 2.6, 4.4 mg
Iludioxonil/L.
Test design: 7 Iish per concentration, control and aquarium.
Sampling: Mortalities and sublethal signs oI toxicity as well as oxygen content, pH and temperature aIter 0, 2-4,
24, 48, 72 and 96 hr. oI exposure.
Statistical analysis: The LC-50 values were calculated according to Berkson, JASA 48 (1953, 569 599. LC-
values were also graphically determined on gausso-logarithmic probability paper.
Test conditions: Static conditions, T 23-24C, pH 7.7 8.4, O
2
58 102. 20 L glass aquaria Iilled with
15 L (36x22x25cm
3
), 16 hr. light, 8 hr. dark.
Results:
AIter 2-4 hr. all Iish were still alive (Table 9.2.1-14).
The treatment related eIIects observed included altered swimming behaviour, loss oI equilibrium and altered
respiratory Iunction. No incidences oI exophtalmus and altered pigmentation were observed.
LC
50
was 1.8 mg Iludioxonil/L aIter 24, 48, 72 and 96 hr. (Table 9.2.1-14)
acute toxicity study with SWITCH 62.5 WG (A 9219 B), (Iludioxonil 25.1 and cyprodinil 37.6) and Lepomis
macrochirus under static conditions.
Concentration
actual mean (mg
Iludioxonil/L)
2-4 hr. 24 hr. 48 hr. 72 hr. 96 hr.
Control 0 0 0 0 0 0 0 0 0 0
0.2 0 0 0 0 0 0 0 0 0 0
0.6 0 0 0 0 0
1.3 0 0 0 0 0
2.6 0 100 - 100 - 100 - 100 -
4.4 0 100 - 100 - 100 - 100 -
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37
Test species: Rainbow trout Oncorhvnchus mvkiss Irom Forellenzucht P. Hohler-Gasser, Zeiningen, Zwitzerland.
Wet weight: mean 0.64 g, range 0.5 to 1.09 g. Length: mean 41 mm, range 40 to 45 mm.
Test concentrations: control, 5, 10, 20, 40, 80 mg A 8207 I/L, nominal concentrations oI Iludioxonil (CGA
173506) were control, 0.124, 0.248, 0.495, 0.990, 1.981. Measured concentrations oI Iludioxonil (CGA 173506)
at hr. 0 were control, 0.105, 0.225, 0.5536, 0.838, 1.681, and at hr. 96 they were control, 0.115, 0.210, 0.393,
0.622, 1.076.
Test design: One replicate with 7 Iish per test concentration and controls were exposed Ior 96 hr. to an aqueous
solution oI A 8207 I under static test conditions.
Sampling: Clinical signs oI toxicity, abnormal behaviour and mortality at 0, 2, 4, 24, 48, 72, 96-hr. pH, dissolved
oxygen concentration, temperature daily.
Statistical analysis: Probit analysis
Test conditions: Static conditions in 15 L aquaria Iilled with 14 L oI the test solutions and control, During the
96-hr. test, pH varied between 6.93 and 7.33, dissolved oxygen varied between 8.82 and 11.57 mg/L or 87 and
114 oI saturation, and temperature between 12.6C and 13.3C, min/max were 12.2C/13.9C.
Results:
No mortalities were observed in the control (Table 9.2.1-16 ).
Sublethal eIIects were observed at all test concentrations, but not in the controls (Table 9.2.1-16 ).
Mortality only occurred at the two highest test concentrations (Table 9.2.1-16 ).
LC
50
, LOEC and NOEC are presented in Table 9.2.1-17.
acute toxicity study with Oncorhvnchus mvkiss under static conditions.
4 hr. 24 hr. 48 hr. 72 hr. 96 hr. Nominal concentration
oI A 8207 I (mg/L)
Control 0 0 0 0 0 0 0 0 0 0
5 0 0 0 7 0 7 0 7 0 7
10 0 7 0 7 0 7 0 7 0 7
20 0 7 0 7 0 7 0 7 0 7
40 0 7 100 - 100 - 100 - 100 -
80 0 7 100 - 100 - 100 - 100 -
* Treatment related eIIects included lethargy, reduced respiration, abnormal behavior and loss oI equilibrium
Table 9.2.1-17: Results Irom acute toxicity study with CELEST 025 FS (A 8207 I) on. Rainbow trout under
static conditions.
Test substance CELEST 025 FS (mg/L)
Nominal concentrations
Fludioxonil (mg as/L)
Nominal conc.
Fludioxonil (mg as/L)
mean measured conc.*
Test object Rainbow trout Oncorhvnchus mvkiss
LC
50
(95 C.L.)-96-hr. 28 0.69# 0.588*
LOEC 5 0.124 0.11*
NOEC (Mortality) 20 0.495 0.47*
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39
Test concentrations: Nominals were 0.0025, 0.01, 0.04, 0.16 and 0.64 mg as/L. Actual measured means were
0.0024, 0.014, 0.0052, 0.249, and 0.348 mg as/L. Negative and solvent (polyethylenglycol) controls were
included.
Test design: Ten Iish per concentration and control, and ten per aquarium. Five nominal test concentrations
separated by a Iactor oI Iour.
Sampling: Mortalities, weight, length, signs oI intoxication and abnormal behaviour.
Statistical analysis: NOEC~TransIormation Iollowed by an approach where a linear regression model with
quadratic and general lack oI Iit, is Iit to the data. The signiIicance oI each term is tested by repeatedly
comparison starting with the Iirst two concentration groups. A second control is compared to the Iirst by a T-test.
Test conditions: 21-day test under Ilow-through conditions in 20 L aquaria Iilled with 15 L test solution each,
light was 16 hr. oI Iluorescent light daily. Temperature was between 14 and 16C, pH between 7.5 and 8.1,
oxygen saturation between 65 and 88.
Results:
The concentrations oI Iludioxonil were measured at regular intervals throughout the 21-day test period (Table
9.2.2-1). At the two highest test concentrations there were problems with the Iormulation oI the stock solutions
(Ilocculation oI the test substance) and consequently there was diIIiculties with the dosing into the tanks.
Table 9.2.2-1: Analytical data oI test concentrations oI Iludioxonil (CGA 173506)
Nominal concentration
(mg as/L)
Day 0
mg as/L
Day 2
mg as/L
Day 7
mg as/L
Day 14
mg as/L
Day 21
mg as/L
Mean
mg as/L ( nom)
0.0025 0.0040 0.0031 0.0018 0.0018 0.0015 0.0024 (96)
0.01 0.018 0.012 0.017 0.014 0.01 0.014 (140)
0.04 0.006 0.005 0.005 0.007 0.003 0.0052 (13)
0.16 0.66 0.057 0.30 0.13 0.10 0.249 (156)
0.64 0.96 0.13 0.23 0.31 0.11 0.348 (54)
Mortalities and symptoms oI intoxication were recorded daily except on Sundays. On day seven 3 Iish at the
0.25 mg as/L reacted with temporary deviant behaviour concerning Iood intake and reaction to external stimuli.
In the solvent control two Iish died because they jumped out oI the aquarium, and in each oI the 0.25 and 0.35
mg as/L treatment concentrations one Iish died Irom jumping out. At the 0.35 mg as/L two Iish died in the
aquaria.
Table 9.2.2-2: Mortalities (x) and other observed eIIects (y) (x-y) in 21-day prolonged toxicity study oI
Iludioxonil (CGA 173506) eIIects on rainbow trout Oncorhvnchus mvkiss.
Mean measured concentrations oI Iludioxonil
Day
Control Solvent
control
0.0024
mg as/L
0.0052
mg as/L
0.014
mg as/L
0.249
mg as/L
0.348
mg as/L
1 0-0 0-0 0-0 0-0 0-0 0-1 0-10
2 0-0 0-0 0-0 0-0 0-0 0-1 0-10
3 0-0 0-0 0-0 0-0 0-0 0-0 0-10
4 0-0 0-0 0-0 0-0 0-0 0-0 0-10
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40
Mean measured concentrations oI Iludioxonil
Day
Control Solvent
control
0.0024
mg as/L
0.0052
mg as/L
0.014
mg as/L
0.249
mg as/L
0.348
mg as/L
5 0-0 0-0 0-0 0-0 0-0 0-0 0-10
7 0-0 0-0 0-0 0-0 0-0 0-3 0-10
8 0-0 0-0 0-0 0-0 0-0 0-0 0-10
9 0-0 0-0 0-0 0-0 0-0 1-0 1-9
10 0-0 0-0 0-0 0-0 0-0 1-0 1-9
11 0-0 2-0* 0-0 0-0 0-0 1-0 2-8
12 0-0 2-0 0-0 0-0 0-0 1-0 2-8
14 0-0 2-0 0-0 0-0 0-0 1-0 2-8
15 0-0 2-0 0-0 0-0 0-0 1-0 2-8
16 0-0 2-0 0-0 0-0 0-0 1-0 3-7
17 0-0 2-0 1-0 0-0 0-0 1-0 3-7
18 0-0 2-0 1-0 0-0 0-0 1-0 3-7
19 0-0 2-0 1-0 0-0 0-0 1-0 3-7
21 0-0 2-0 1-0 0-0 0-0 1-0 3-7
Table 9.2.2-3: EIIects on weight and length gain Irom investigation oI prolonged toxicity oI Iludioxonil to
rainbow trout Oncorhvnchus mvkiss.
Mean measured
concentration
(mg Iludioxonil/L)
Rate oI weight gain*
Mean SD
Rate oI length gain*
Mean SD
Blank 0.0248 0.0041 0.0074 0.0058
Vehicle 0.0285 0.0052 0.0087 0.0023
0.0024 0.0272 0.0064 0.0081 0.0022
0.014 0.0261 0.0061 0.0088 0.0016
0.0052 0.0276 0.0051 0.0093 0.0013
0.249 0.0245 0.0072 0.0081 0.0023
0.348 -0.0123 0.0073 - 0.0012 0.0016
* Mean values and standard deviation calculated by RMS only raw data were provided in the report
Table 9.2.2-4: NOEC Ior Iludioxonil (CGA 173506) to rainbow trout Oncorhvnchus mvkiss.
Test substance Fludioxonil (mg as/L)
Test duration 21 days
LC
50
~ 0.348
LOEC (Mortality, weight, length) 0.249
#
NOEC (Mortality, weight, length) 0.249
#
NOEC (behaviour) 0.014*
#
RMS does not Iind these values reliable due to Iluctuating test concentrations
*The NOEC was based on temporary sublethal eIIects (deviating Iood intake and reaction to external stimulation) on day
seven in 0.249 mg as/L.
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42
The purpose oI the study was to determine threshold levels oI lethal eIIects and the NOEC oI CGA 173506 tech.
on rainbow trout Oncorhvnchus mvkiss.
Test substance: Fludioxonil CGA 173506, Batch No. P910007, purity 96.4.
was 47mm (37 57mm), Mean weight oI Iish was 1.14g (0.43 1.93g). Fish were Ireeze branded 24h beIore the
start oI exposure.
Test concentrations: Nominals were 0.0025, 0.01, 0.04, 0.16 and 0.64 mg as/L. Mean results Irom analytical data
oI test concentrations were 0.0023, 0.0088, 0.010, 0.306, and 1.155 mg Iludioxonil/L. Negative and solvent
(dimethylsulIoxide) controls were included.
Statistical analysis: NOEC ~ TransIormation Iollowed by an approach where a linear regression model with
quadratic and general lack oI Iit is Iit to the data. The signiIicance oI each term is tested by repeatedly
comparison starting with the Iirst two concentrations groups. A second control is compared to the Iirst by a T-
test.
light was 16 hours oI Iluorescent light daily. Temperature ranged Irom 14 to 16C, pH ranged Irom 7.5 to 8.1,
oxygen saturation ranged Irom 65 to 89.
Results:
The results oI the analyses oI Iludioxonil test concentrations are presented in Table 9.2.2-5. It is seen that the two
lowest test concentrations were quite constant and only slightly lower than the nominal concentrations. The 0.04
mg Iludioxonil/L nominal concentration varied signiIicantly over time and as an average it was only 25 oI the
nominal. The two highest concentrations were constantly higher than their nominals, and their averages were 191
and 180 higher than the nominals, respectively. Observations oI mortalities and other observed eIIects are
presented in Table 9.2.2-6 and mean results Ior weight and length gain in Table 9.2.2-6. The Iish that died in the
0.0023 mg Iludioxonil/L test concentration died because it was wounded by maintenance, and the Iish that died
at the 0.010 mg Iludioxonil/L test concentration jumped out oI the aquarium. The Iirst Iish that died at the
second highest test concentration jumped out oI the aquarium, the other died without accident. All Iive Iish that
died at the highest test concentration died without accidents.
Table 9.2.2-5: Analytical data oI test concentrations oI Iludioxonil
mg Iludioxonil/L Nominal concentration
(mg Iludioxonil/L)
Day 0
Day 2
Day 7 Day 14 Day 21
Mean
mg Iludioxonil/L
( nom)
0.0025 0.0024 0.0022 0.0015 0.0023 0.0029 0.0023 (91)
0.01 0.009 0.010 0.006 0.008 0.011 0.0088 (88)
0.04 0.033 0.005 0.005 0.008 0.004 0.010 (25)
0.16 0.19 0.24 0.30 0.36 0.44 0.306 (191)
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Sun Aug 18 05:04:43 CEST 2013
43
0.64 1.53 0.92 0.94 1.39 1.47 1.155 (180)
Table 9.2.2-6: Mortalities (x) and other observed eIIects (y) presented as (x-y) in 21-day prolonged toxicity
study oI Iludioxonil CGA173506 eIIects on rainbow trout Oncorhvnchus mvkiss.
Mean measured
concentration
Control
Solvent 0.0023 0.0088 0.010 0.306 1.155
Nominal conc. Control Solvent 0.0025 0.01 0.04 0.16 0.64
Day mg Iludioxonil/L
1
0-0 0-0 0-0 0-0 0-0
0-5 0-10
2 0-0 0-0 1-0 0-0 0-0 0-5 0-10
3 0-0 0-0 1-0 0-0 0-0 0-3 0-10
4 0-0 0-0 1-0 0-0 0-0 0-3 0-10
5 0-0 0-0 1-0 0-0 0-0 0-3 0-10
7 0-0 0-0 1-0 0-0 0-0 0-5 1-9
8 0-0 0-0 1-0 0-0 0-0 0-3 5-5
9 0-0 0-0 1-0 0-0 0-0 0-1 5-5
10 0-0 0-0 1-0 0-0 0-0 0-0 5-5
11 0-0 2-0 1-0 0-0 1-0 1-0* 5-5
12 0-0 2-0 1-0 0-0 1-0 1-0 5-5
14 0-0 2-0 1-0 0-0 1-0 1-3 5-5
15 0-0 2-0 1-0 0-0 1-0 2-2 5-5
16 0-0 2-0 1-0 0-0 1-0 2-2 5-5
17 0-0 2-0 1-0 0-0 1-0 2-2 5-5
18 0-0 2-0 1-0 0-0 1-0 2-2 5-5
19 0-0 2-0 1-0 0-0 1-0 2-2 5-5
21 0-0 2-0 1-0 0-0 1-0 3-2 5-5
Mean measured
concentration
(mg Iludioxonil/L)
Mean SD
Mean SD
Blank 0.0248 0.0041 0.0082 0.0017
Vehicle 0.0271 0.0061 0.0098 0.0018
0.0023 0.0281 0.0076 0.0097 0.0024
0.0088 0.0281 0.0057 0.0096 0.0020
0.010 0.0290 0.0132 0.0100 0.0041
0.306 0.0072 0.0099
#
0.0033 0.0023
#
1.155 - 0.0135 0.0029
#
0.0007 0.0007
#
* Mean values and standard deviation calculated by RMS only raw data were provided in the report
#
Statistically signiIicantly diIIerent Irom control
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45
The purpose oI the study was to evaluate the prolonged toxicity oI CGA 173506 tech. to rainbow trout
Test substance: Fludioxonil CGA 173506, Batch No. P910007, purity 96.4.
was 51mm (45 59mm), Mean weight oI Iish was 1.42g (0.84 2.18g). Fish were Ireeze branded 24h beIore the
start oI exposure.
Test concentrations: Nominals were 0.000039, 0.000156, 0.000625, 0.0025, and 0.01 mg as/L. Negative and
solvent (polyethylenglycol) controls were included.
Statistical analysis: NOEC~TransIormation Iollowed by an approach where a linear regression model with
quadratic and general lack oI Iit is Iit to the data. The signiIicance oI each term is tested by repeated comparison
starting with the Iirst two concentrations groups. A second control is compared to the Iirst by a T-test.
light was 16 hours oI Iluorescent light daily. Loading oI Iish was 0.16 g/L/day. Temperature was between 14 and
15C, pH was between 7.5 and 8.1, and oxygen saturation was between 68 and 100 (gentle aeration).
Results:
The results Irom the analysis oI the actual concentration oI the test compound are presented in Table 9.2.2-9. The
measured concentrations were only determined Ior the highest test concentration, i.e. nominal 0.01 mg
Iludioxonil/L.
There were no signs oI intoxication, abnormal behaviour or mortalities recorded during the experiment (Table
9.2.2-10) and there were no eIIects on weight or length gain (Table 9.2.2-11). ThereIore the NOEC is equal to
the highest concentration tested (Table 9.2.2-12).
Table 9.2.2-9: Analytical data oI test concentrations oI Iludioxonil.
Measured Iludioxonil concentration (mg Iludioxonil/L)
(mg Iludioxonil/L)
Day 0
Day 7 Day 14 Day 21 Mean
mg/L ( nom)
0.000039
0.000156
0.000625
0.0025
Not analysed
0.01 0.0063 Not det. 0.0051 0.0084 0.0066 (66)
Table 9.2.2-10: Mortalities (x) and other observed eIIects (y) (x-y) in 21-day prolonged toxicity study oI
Iludioxonil CGA173506 eIIects on rainbow trout Oncorhvnchus mvkiss.
Nominal concentration (mg Iludioxonil/L)
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46
Day
Control
Solvent
control
0.000039 0.000156 0.000625 0.0025 0.01
1
0-0 0-0 0-0 0-0 0-0 0-0 0-0
2 0-0 0-0 0-0 0-0 0-0 0-0 0-0
3 0-0 0-0 0-0 0-0 0-0 0-0 0-0
4 0-0 0-0 0-0 0-0 0-0 0-0 0-0
5 0-0 0-0 0-0 0-0 0-0 0-0 0-0
7 0-0 0-0 0-0 0-0 0-0 0-0 0-0
8 0-0 0-0 0-0 0-0 0-0 0-0 0-0
9 0-0 0-0 0-0 0-0 0-0 0-0 0-0
10 0-0 0-0 0-0 0-0 0-0 0-0 0-0
11 0-0 0-0 0-0 0-0 0-0 0-0 0-0
12 0-0 0-0 0-0 0-0 0-0 0-0 0-0
14 0-0 0-0 0-0 0-0 0-0 0-0 0-0
15 0-0 0-0 0-0 0-0 0-0 0-0 0-0
16 0-0 0-0 0-0 0-0 0-0 0-0 0-0
17 0-0 0-0 0-0 0-0 0-0 0-0 0-0
18 0-0 0-0 0-0 0-0 0-0 0-0 0-0
19 0-0 0-0 0-0 0-0 0-0 0-0 0-0
21 0-0 0-0 0-0 0-0 0-0 0-0 0-0
(mg Iludioxonil/L)
Mean SD
Mean SD
Blank 0.0158 0.0005 0.0081 0.0026
Vehicle 0.0165 0.0033 0.0081 0.0010
0.000039 0.0133 0.0082 0.0091 0.0038
0.000156
0.0148 0.0072 0.0070 0.0023
0.000625 0.0157 0.0069 0.0081 0.0130
0.0025 0.0147 0.0071 0.0080 0.0017
0.01 0.0136 0.0056 0.0084 0.0017
* Mean values and standard deviation calculated by RMS only raw data were provided in the report.
Table 9.2.2-12: NOEC Ior Iludioxonil to rainbow trout Oncorhvnchus mvkiss.
Test substance Mean measured mg Iludioxonil/L Nominal mg Iludioxonil/L)
Test object Oncorhvnchus mvkiss Oncorhvnchus mvkiss
Test duration 21 days 21 days
LOEC ~ 0.0066 ~0.01
NOEC (Mortality) 0.0066 0.01
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48
Results:
Actual means oI the test concentrations were calculated Irom analysis oI the stock solutions (based on a
measured mean oI 29 oI the nominal concentration, however actual concentrations Iluctuated between 7 and
63 oI nominal), they were 0.00000029, 0.0000011, 0.0000046, 0.000018, and 0.000073 mg Iludioxonil/L, i.e.
lower than the intended nominal concentrations.
There were neither observations oI dead Iish, nor oI abnormal behaviour throughout the study period.
There were eIIects on the rate oI length and weight gain at concentrations higher than 0.0000046 mg
Iludioxonil/L (Table 9.2.2-13), where rate oI length gain and rate oI weight gain were signiIicantly lower at the
two highest concentrations.
(mg Iludioxonil/L)
Mean SD
Mean SD
Blank 0.0255 0.0104 0.0076 0.0023
Vehicle 0.0261 0.0136 0.0074 0.0039
0.00000029 0.0264 0.0050 0.0069 0.0020
0.0000011 0.0285 0.0068 0.0080 0.0026
0.0000046 0.0264 0.0052 0.0073 0.0024
0.000018 0.0152 0.0076
#
0.0036 0.0014
#
0.000073 0.0092 0.0048
#
0.0024 0.0011
#
* Mean values and standard deviation calculated by RMS only raw data were provided in the report.
#
SigniIicantly lower than control
Table 9.2.2-14: NOEC Ior Iludioxonil to rainbow trout Oncorhvnchus mvkiss.
Test substance Iludioxonil (mg/L)
Test duration 21 days
LOEC (Mortality) ~0.000073
LOEC (Sublethal) 0.000018
LOEC and NOEC sublethal are based on rate oI length and weight gain (which are reduced by approximately 40 at
0.000018 and 65 at 0.000073 mg/L - calculated by RMS).
The study was in accordance with the guideline except Ior the length oI the Iish, which is not believed to have
inIluenced the results. However the concentrations oI the test substance were too low to determine direct Irom
the test solutions. ThereIore they were calculated on the basis oI analyses oI the stock solutions, which in terms
Iluctuated and on average amounted to only 29 oI nominal concentrations. Initially the NotiIier claimed that
the solvent (DMSO) inIluenced the study by increasing the toxicity oI the test substance, however this was later
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50
behaviour, Iish lying on side or back at the bottom, strong ventilation, tumbling during swimming, changed body
colour, indigestion)
Statistical analysis: ANOVA, Williams-Test
Test conditions: 21 days, T 13.3 14.5C, 16 hr. light/8 hr. dark. For the test ten Iish per test concentration and
control were held in aquaria, which were 0.54x0.29x0.395m
3
. pH ranged Irom 7.3 to 8.2. Oxygen ranged Irom
6.3 to 10.2 mg/L
Results:
Fish treated with the two highest test concentrations showed signs oI intoxication aIter 2 hr., and aIter 4 days the
Iirst Iish died at the nominal 1.0 mg SWITCH 62.5 WG /L treatment. At the end oI the test, 9 Iish had died at 1.0
mg SWITCH 62.5 WG /L and 10 had died at 3.2 mg/L nominal test concentrations Table 9.2.2-12.
At all the lower concentrations and the control no Iish died and no signs oI intoxication were observed (Table
9.2.2-15).
Length and body weight oI surviving Iish were only signiIicantly lower than in the control in the Iish that
survived at the second highest test concentration (Table 9.2.2-16). In all concentrations lower than 1.0 mg
SWITCH 62.5 WG/L length and body weight were essentially identical with the length and bodyweight oI the
control.
Table 9.2.2-15: Mortalities and observed signs oI intoxication in a 21 day study exposing rainbow trout
Oncorhvnchus mvkiss to SWITCH 62.5 WG (25.0 Iludioxonil (CGA 173506) and 37.5 cyprodinil (CGA
219417)).
Nominal
concentration
(mg/L)
2 hr.
D/S*
Day 4
D/S*
Day 7
D/S*
Day 11
D/S*
Day 14
D/S*
Day 18
D/S*
Day 21
D/S*
Control 0/0 0/0 0/0 0/0 0/0 0/0 0/0
0.01 0/0 0/0 0/0 0/0 0/0 0/0 0/0
0.032 0/0 0/0 0/0 0/0 0/0 0/0 0/0
0.10 0/0 0/0 0/0 0/0 0/0 0/0 0/0
0.32 0/0 0/0 0/0 0/0 0/0 0/0 0/0
1.0 0/10 2/10 2/10 3/10 4/10 8/10 9/10
3.2 0/10 0/10 0/10 3/10 4/10 9/10 10/10
* D/S Dead / Symptoms oI intoxication
Table 9.2.2-16: EIIects on weight and length gain Irom investigation oI prolonged toxicity oI SWITCH 62.5 WG
to rainbow trout Oncorhvnchus mvkiss.
(mg/L)
weight gain* length gain*
Control 246 130
0.01 262 131
0.032 277 133
0.10 277 131
0.32 264 133
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GLP: Yes
The purpose oI the study was to evaluate the eIIects oI Iludioxonil on early liIe-stage development oI Iathead
minnows, Pimephales promelas. Time to hatch, hatching success, survival, and growth were evaluated.
Test substance: Fludioxonil (CGA 173506), Batch No. P905004/5, purity 95.4.
Test species: Fathead minnow, Pimephales promelas 4-5 hr. old Irom cultures maintained at WildliIe
International Ltd. Easton, Maryland.
Test concentrations: Nominal test concentrations were 0.0094, 0.019, 0.038, 0.075, and 0.150 mg as/L. Control
and solvent (acetone) controls were included. Mean measured concentrations were 0.010, 0.020, 0.039, 0.077,
and 0.144 mg as/L.
Test design: Fathead minnow embryos were exposed to a geometric series oI Iive test concentrations, a solvent
control, and a negative control. Two replicate test chambers were maintained in each treatment and control
group, with two incubation cups in each chamber. Each incubation cup contained 20 embryos, resulting in a total
oI 40 embryos per replicate, and 80 embryos per treatment. The exposure included a 5-day embryo hatching
period, and a 28-day post hatch juvenile growth period.
Sampling: Observations oI mortality and other clinical signs oI toxicity were made daily during the test. Time to
hatch, hatching success, growth, and survival were monitored.
Statistical analysis: Normality by chi-square or Kolmogorov-Smirnov test, homogeneity Ior variance by
Bartlett`s test, Control groups were compared by Student`s T-test. BonIerronni`s test or SheIIe`s multiple
comparison test were used to evaluate diIIerences between treatment and control means. Alternatively Kruskal-
Wallis test was used to evaluate diIIerences between the treatment and control groups.
Test conditions: Flow-through conditions in 9 L glass aquaria with 7.5 L oI test solution. A photoperiod oI 16 hr.
oI light at 269 lux and 8 hr. oI darkness with a 30 minutes transition period. During the test the temperature
ranged Irom 24.6 to 25.3 C, pH ranged Irom 7.9 8.4, and dissolved oxygen Irom 5.2 8.2 mg/L.
Results:
There were no apparent treatment related signs oI toxicity or abnormal behaviour observed in the 0.010, 0.020,
and 0.039 mg as/L treatments. A higher proportion oI Iish exposed to 0.077 and 0.144 mg as/L appeared weak
and small compared to the control Iish. There were no signiIicant diIIerences with regard to hatching success or
time to hatch between the control and treatment groups (Table 9.2.2-17). Post hatch survival was not
signiIicantly diIIerent between the control groups and the three lowest treatment concentration groups. Post
hatch survival was signiIicantly reduced in the 0.077 and the 0.144 mg as/L treatment groups (Table 9.2.2-18).
The statistically deIined NOEC Ior survival and growth was 0.039 mg as/L (Tables 9.2.2-18 and 9.2.2-19). The
LOEC was 0.077 mg as/kg based on a statistically signiIicant reduction in survival at that concentration.
Table 9.2.2-17: Hatching success oI Iathead minnow Pimephales promelas exposed to Iludioxonil (CGA
173506) in an early liIe-stage toxicity test.
Measured concentration
(mg as/L)
Number oI eggs Number hatched Percent hatching success
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53
(mg as/L)
Number oI eggs Number hatched Percent hatching success
Control 79 68 86
Solvent control 76 67 88
0.010 79 65 82
0.020 77 69 90
0.039 77 62 81
0.077 78 67 86
0.144 75 63 84
Table 9.2.2-18: Survival oI Iathead minnow Pimephales promelas exposed to Iludioxonil (CGA 173506) Ior 28
days post hatch in an early liIe-stage toxicity test.
Measured concentration (mg as/L) Initial number oI larvae Percent survival
Control 68 88
Solvent control 67 87
0.010 65 89
0.020 69 88
0.039 62 87
0.077 67 70*
0.144 63 65*
* Statistically signiIicant
Table 9.2.2-19: Total length and dry weight oI Iathead minnow Pimephales promelas exposed to Iludioxonil
(CGA 173506) at the end oI the 28-day post hatch observation period in an early liIe-stage toxicity test.
(mg as/L)
Number oI
surviving larvae
Length (mm)
mean + SD
Dry weight (mg)
mean + SD
Control 60 19.2 + 2.9 12.7 + 4.6
Solvent control 58 19.6 + 2.6 11.8 + 3.5
0.010 58 20.1 + 2.9 14.1 + 5.0
0.020 61 19.0 + 2.6 12.5 + 4.5
0.039 55 18.5 + 3.1 12.0 + 5.4
0.077 47 17.1 + 2.9* 9.8 + 3.9*
0.144 41 15.8 + 2.3* 8.2 + 3.1*
* Statistically signiIicant
Table 9.2.2-20: NOEC Ior Iathead minnow Pimephales promelas exposed to Iludioxonil (CGA 173506) in an
early liIe-stage toxicity test.
Test object Pimephales promelas
Test duration 5-day embryo hatching period 28-day post
hatch period
LOEC 0.077
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55
Test design: Three tanks (two treatments and one control. Fish were continuously exposed to
14
C-pyrrolering
labelled Iludioxonil (CGA 173506) at a concentration oI 0.01 mg/L Ior 28 days in a dynamic Ilow-through
system and thereaIter the depuration oI radioactivity Iollowed in untreated water Ior 14 days.
Sampling: Radioactivity in edible and non-edible parts oI Iish at day 0, 1, 3, 7, 14, 21, and 28 oI the exposure
phase, and at days 1,2, 5, 7, 9, and 14 oI the depuration phase. Analysis oI the test substance and metabolites in
Iish at day 8 and 28 oI the exposure phase.
Statistical analysis: Only calculations stated
Test conditions: Flow-through in three aerated temperature controlled 14-hr. twilight illuminated 100 L clear
glass aquaria. Initial loading oI Iish was 0.5 g/L based on 24-hr. Ilow-through oI test solution. During the test
temperature was between 19.5 and 20.5C, average was 20.2C, percentage oxygen saturation was between 81
and 118, average was 96, and pH was between 7.38 and 7.66, average was 7.54.
Results:
Prior to and during the experiment, all Iish showed normal behaviour and no treatment related symptoms. No
mortality was observed in the two treated tanks and in the control tank. The average level oI radioactivity in
water during the exposure phase was 9.5 + 0.87 g Iludioxonil equivalents/L. In the depuration phase it was 0.2
and 0.7 the two Iirst days respectively, thereaIter it was app. 0, range 0.1 to 0.1). The nature oI the radioactivity
in water was 96.8 as Iludioxonil. The nature oI radioactivity in Iish tissues was at the end oI incubation 87.6
as Iludioxonil in edible parts, and 28.0 as Iludioxonil in non-edible parts. The rest was either in the Iorm oI
metabolites, non-extractables, not analysed or unresolved background.
Table 9.2.3-1: Level oI radioactivity in Iish tissues.
Period Days Edibles Non-edibles Total
(mg equivalents/kg)
0 0.073 0.130 0.098
1 0.315 2.566 1.384
3 0.290 3.690 1.815
7 0.280 4.187 2.132
14 0.441 5.645 2.878
21 0.701 8.449 4.183
Accumulation
28 0.451 6.232 2.987
29 0.470 1.228 0.828
30 0.235 0.316 0.272
33 0.042 0.421 0.218
35 0.041 0.227 0.128
37 0.022 0.293 0.152
Depuration
42 0.037 0.213 0.117
Average at steady state 0.531 6.775 3.349
Table 9.2.3-2: Regression output Ior uptake and depuration data
Edibles Non-edibles Total
Accumulation
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56
R
2
0.51329 0.84025 0.90815
K
u
13.99 153.3 83.0
K
d
0.2481 0.20458 0.22731
C
w
(g equiv./L) 9.15 9.15 9.15
T
80
(ln 5/K
d
) |d| 6.4 7.8 7.1
T
95
(ln 20/K
d
) |d| 12.1 14.7 13.2
Depuration
R
2
0.9215 0.99344 0.99246
K
e
0.33372 1.60245 1.2531
C
o
(mg equiv./L) 0.5053 6.2293 2.9829
DT
50
(ln 2/K
e
)

|d| 2.1 0.4 0.6
DT
90
(ln 10/K
e
)

|d| 6.9 1.4 1.8
K
u
uptake constant, K
d
depuration constant Irom Iish tissue, K
e
elimination constant Irom Iish tissue, C
w
mean water
concentration during uptake phase, C
o
concentration in Iish tissue at the beginning oI depuration phase, T
80
the time to
reach 80 oI the steady state concentration, T
95
the time to reach 95 oI the steady state concentration, DT-50 the halI-
liIe Ior depuration oI the test compound Irom tissues, DT-90 the time Ior 90 depuration oI the test compound Irom
tissues.
Table 9.2.3-3: Bioconcentration Iactors in Iish (juvenile bluegill sunIish, Lepomis macrochirus) based on
14
C-
labelled Iludioxonil (CGA 173506).
Measured (C
tissue
/C
water
) 58 741 366
Calculated (k
u
/k
d
) 56 749 365
The study was acceptable and in agreement with the guideline. The conclusion was that the BCF Ior total Iish
was 366 (Table 9.2.3-3) and that Iludioxonil was eliminated soon aIter end oI the exposure. DT
50
and DT
90
Ior
total Iish were 0.6 and 1.8 days respectively (Table 9.2.3-1).
B.9.2.4 ACUTE TOXICITY TO AQUATIC INVERTEBRATES (ANNEX IIA 8.2.4; ANNEX IIIA 10.2.1)
REFERENCE: Annex IIA, 8.2.4.1
Report:
Surprenant, D.C. (1990a). (CGA 173506 Technical) - Acute toxicity to daphnids (Daphnia magna) under Ilow-
through conditions. Syngenta Crop Protection Unpublished report CGA173506/0059 Springborn Laboratories
Inc., Wareham, Massachusetts, USA, Unpublished report No. 89-05-2990.
Guidelines:
US EPA FIFRA Guideline No. 72-2, Acute Toxicity Test Ior Freshwater Aquatic Invertebrates
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57
GLP: Yes
The purpose oI the study was to determine the acute toxicity oI Iludioxonil (CGA 173506) to the Cladoceran
Daphnia magna.
Test substance: Fludioxonil (CGA 173506) Technical, Batch No. P805002, FL No. 881677. Purity was 97.5.
Test species: Daphnia magna. The daphnids used were 24 hr., they were obtained Irom laboratory cultures
maintained at Springborn Laboratories Inc, Wareham, Massachusetts.
Test concentrations: Nominal were 0.13, 0.22, 0.36, 0.60 and 1.0 mg Iludioxonil /L. Mean measured were 0.12,
0.21, 0.31, 0.54, and 0.76 mg Iludioxonil /L. Control and solvent control were included. Nominal concentrations
were based on range-Iinding test.
Test design: Two replicate test chambers per treatment and control groups were maintained with 10 juvenile
daphnids in each test chamber Ior a total oI 20 daphnids per concentration. Sampling: Immobilisation and
behaviour at test initiation and aIter 24 and 48 hr. oI exposure.
Statistical analysis: Moving average angle analysis, probit analysis, and non-linear interpolation with 95
conIidence intervals calculated by binomial probability.
Test conditions: Flow-through. During the test pH varied Irom 8.1 to 8.2, dissolved oxygen varied between 7.3
and 8.5 mg/L, and temperature was held at 21C.
Results:
The results presented in table 9.2.4-1 were based on the mean measured concentrations oI Iludioxonil that appear
in the leIt column oI Table 9.2.4-1. AIter 48 hr. oI exposure and average oI 95, 60 and 25 oI the daphnids were
immobilised in the three highest concentrations tested (Table 9.2.4-1). All oI the surviving daphnids at the two
highest test concentrations, and several oI the daphnids exposed to the 0.31 mg Iludioxonil/L exhibited lethargic
behaviour. There was less than 15 immobilisation and no abnormal behaviour observed in the 0.12 and 0.21
mg Iludioxonil/L treatments. The EC
50
values are reported in Table 9.2.4-2.
The NOEC could not be unequivocally determined due to the occurrence oI slight mortality at the two lowest
test concentrations. However, based on the behaviour oI surviving daphnia at the 0.12 and 0.21 mg Iludioxonil/L
levels, and the Iact that some incidental mortality Irequently occurs with young daphnia the daphnia are
considered to be unaIIected by exposure to the test material at the two lowest concentrations tested.
Table 9.2.4-1: Concentrations tested and corresponding cumulative percent oI immobilised organisms during the
48-hr. exposure oI daphnids Daphnia magna to Iludioxonil.
Cumulative percent oI immobilised organisms
24-hr. 48-hr.
Mean measured
concentration
(mg as/L)
A B Mean A B Mean
0.76 30 70 50* 90 100 95*
0.54 10 10 10* 50 70 60*
0.31 10 0 5 40 10 25*
0.21 0 0 0 10 10 10
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Sun Aug 18 05:04:43 CEST 2013
58
Cumulative percent oI immobilised organisms
24-hr. 48-hr.
Mean measured
concentration
(mg as/L)
A B Mean A B Mean
0.12 0 0 0 10 20 15
Solvent control 0 0 0 0 0 0
control 0 0 0 10 0 5
* surviving daphnids exhibited signs oI being aIIected
Table 9.2.4-2: Acute toxicity oI Iludioxonil technical to Daphnia magna.
Test object Daphnia magna
Test duration 24 hr 48 hr
EC
50
(immobilisation) 0.82 (0.67 1.4) 0.40 (0.33 0.50)
The study is acceptable, and in accordance with the guideline. The 48 h EC
50
Ior the eIIect oI Iludioxonil (CGA
173506) on Daphnia magna was 0.40 mg as/L.
REFERENCE: Annex KIIA, 8.2.4.1
Report:
Holmes, C.M. and Swigert, J.P. (1993c). CGA 173506, a 48-hr. Ilow through acute toxicity test with the
cladoceran (Daphnia magna), Syngenta Crop Protection Unpublished Report No. CGA173506/0259. WildliIe
International Ltd. Easton, Maryland, USA, unpublished report No. 108A-133.
Guidelines:
US EPA FIFRA Guideline No. 72-2, Acute Toxicity Test Ior Freshwater Aquatic Invertebrates.
GLP: Yes
The purpose oI the study was to determine the acute toxicity oI Iludioxonil to the Cladoceran Daphnia magna
under Ilow-through conditions.
Test substance: Fludioxonil (CGA 173506) Technical, ID No: FL-891552 ARS-16651. Batch code: P905004/5.
Purity: 95.4.
Test species: Daphnia magna. The daphnids used were 24 hr., they were obtained Irom WildliIe International
Ltd. Cultures, 8651 Brooks Drive, Easton, Maryland 21601, USA.
Test concentrations: Nominal were 0.65, 1.1, 1.8, 3.0 and 5.0 mg Iludioxonil /L. Mean measured were 0.50,
0.83, 1.1, 1.6, and 1.9 mg Iludioxonil /L. Control and solvent control were included. The solvent concentration in
W
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Sun Aug 18 05:04:43 CEST 2013
59
the treatment and solvent control groups was 1.0 mL acetone /L. Nominal concentrations were based on range-
Iinding test.
Test design: Daphnids were exposed to a geometric series oI Iive test concentrations, a solvent control and a
negative control. Two replicate test chambers were maintained in each treatment and control, with 10 daphnids
in each test chamber Ior a total oI 20 daphnids per concentration.
Sampling: Immobilisation and behaviour at 3.5, 24 and 48 hr.
Statistical analysis: Moving average method, Probit analysis, and binomial probability with non-linear
interpolation.
Test conditions: Flow-through. During the test pH varied Irom 8.0 to 8.3, dissolved oxygen varied between 8.6
and 9.0 mg/L, and temperature was between 19.7 and 20C.
Results:
The results presented in table 9.2.4-3 were based on the mean measured concentrations oI Iludioxonil that appear
in the leIt column oI Table 9.2.4-1. The low mean measured concentrations at the three highest nominal
concentrations were unavoidable because testing was carried out above the limit oI solubility. At the lowest
Iludioxonil concentration tested (0.50 mg as/L) 45 oI the test organisms appeared normal, 40 oI the test
organisms were lethargic, and 15 were immobile or dead. OI the daphnids exposed to 0.83 mg as/L 10
appeared normal, 45 were lethargic and another 45 were immobile or dead. In the 1.1 mg as/L treatment
group 5 oI the daphnids appeared normal, 35 were lethargic, and 60 were immobile or dead. OI those
exposed to 1.6 mg as/L 20 were lethargic and 80 were immobile or dead. At the highest concentration tested,
1.6 mg as/L, all daphnids were immobile or dead by the end oI the 48 hr. exposure period (Table 9.2.4-3). Based
on these data the EC
50
Ior Iludioxonil was calculated to 0.90 mg as/L, the LOEC was 0.50 mg as/L, i.e. the
lowest concentration tested, and NOEC was not determined (Table 9.2.4-4).
Table 9.2.4-3: Cumulative percent oI immobilised organisms and treatment related eIIects during the 48-hr.
exposure oI daphnids Daphnia magna to Iludioxonil (CGA 173506)
24 hr. 48 hr.
Mean measured
concentration
(mg/L)
EIIects Number
immobile
Number
dead
EIIects Number
immobile
Number
dead
Immobile
and dead
()
Negative control 10 AN
10 AN
0
0
0
0
9 AN
10 AN
0
0
1
0
5
Solvent control 10 AN
10 AN
0
0
0
0
10 AN
10 AN
0
0
0
0
0
0.50 10 AN
9 AN
0
0
0
1
8 C
9 AN
2
0
0
1
15
0.83 10 AN
9 AN
0
0
0
0
2 AN
9 C
1
1
7
0
45
1.1 7 AN
6 AN
0
0
0
0
4 C
3 C; 1
AN
6
2
0
4
60
1.6 9 AN
7 AN
0
0
0
0
1 C
3 C
9
7
0
0
80
1.9 6 AN 0 0 N/A 7 3 100
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60
24 hr. 48 hr.
Mean measured
concentration
(mg/L)
EIIects Number
immobile
Number
dead
EIIects Number
immobile
Number
dead
Immobile
and dead
()
4 AN 0 2 N/A 6 4
AN appeared normal
C lethargic
N/A not applicable
Table 9.2.4-4: Acute toxicity oI Iludioxonil (CGA 173506) technical to Daphnia magna.
Test duration 48 hr.
EC
50
(immobilisation) (95 C.I.) 0.90 (0.75 1.0)
LOEC 0.50
NOEC 0.50
C.I. conIidence interval
The study was acceptable. The concentration oI solvent (acetone) in the solvent control and treatments did not
inIluence the results oI the experiment. The 48 h EC
50
oI Iludioxonil Ior immobilisation oI daphnids was 0.90
mg as/L, LOEC was 0.50 mg as/L. NOEC was not determined, but it was lower than 0.50 mg as/L (Table 9.2.4-
4).
Report:
Holmes, C.M. and Swigert, J.P. (1993d). CGA 173506 - A 96-hr. Ilow through acute toxicity test with the
saltwater mysid (Mvsidopsis bahia). Syngenta Crop Protection Unpublished Report No. CGA173506/0256.
WildliIe International Ltd. Easton, Maryland, USA, unpublished report No. 108A-134.
Guidelines:
US EPA FIFRA Guideline No. 72-3, Acute Toxicity Test Ior Estuarine and Marine Aquatic Organisms
GLP: Yes
The purpose oI the study was to evaluate the acute toxicity oI Iludioxonil to the saltwater mysid Mvsidopsis
bahia during a 96-hr. exposure period under Ilow-through conditions.
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61
Test substance: Fludioxonil (CGA 173506) Technical, ID No: FL-891552 ARS-16651. Batch No. P905004/5.
Purity: 95.4.
Test species: Juvenile saltwater mysids Mvsidopsis bahia. The mysids used were 24 hr., they were obtained
Irom WildliIe International Ltd. Cultures, 8651 Brooks Drive, Easton, Maryland 21601, USA.
Test concentrations: Nominal test concentrations were 0.10, 0.17, 0.29, 0.48, and 0.80 Iludioxonil/L. Mean
measured test concentrations were 0.075, 0.15, 0.24, 0.37, and 0.63 mg Iludioxonil/L. The test included negative
(saltwater) controls and solvent (acetone) controls. Nominal concentrations were based on range-Iinding test.
Test design: Saltwater mysids were exposed to a geometric series oI Iive test concentrations, a solvent control,
and a negative control. Two replicate test chambers were maintained in each treatment and control group, with
10 mysids in each test chamber.
Sampling: Observations oI mortality and other clinical signs oI toxicity were made approximately 4, 24, 48, 72,
and 96 hr. aIter test initiation. Cumulative percent mortalities in the treatment groups were use to calculate LC
50
values. The NOEC was determined by visual examination oI mortality and clinical observation data.
Statistical analysis: LC
50
values and 95 conIidence intervals were calculated by probit analysis, the moving
average method, or by binomial probability.
Test conditions: The measured temperature was between 24.8 and 25.2C, pH ranged Irom 8.2 to 8.3, and the
oxygen concentration was between 6.0 and 7.4 mg/L. The photoperiod was 16 hr. oI light and 8 hr. oI darkness.
Natural seawater adjusted to 2.5 salinity was used.
Results:
Average measured concentrations (day 0 and 4) oI Iludioxonil are presented in the leIt column oI Table 9.2.4-5.
Observations oI mortality and other signs oI toxicity are presented in Table 9.2.4-5. With the exception oI two
incidental deaths in the solvent control mysids in the controls appeared healthy and normal throughout the test.
With the exception oI one incidental death in the 0.075 mg as/L treatment group mysids was unaIIected in this
group. In the 0.15 mg as/L treatment, three mortalities were observed 24 hr. aIter test initiation. No other
mortalities occurred in this group. They were considered incidental because they occurred early in the test and
because they were comparable to the number oI deaths in the solvent control. There did appear to be treatment
related eIIects oI Iludioxonil at all test concentration equal to or greater than 0.24 mg as/L (Table 9.2.4-5) Based
on these data LC
50
LOEC and NOEC values were calculated (Table 9.2.4-6).
Table 9.2.4-5: Cumulative percent mortality and treatment related eIIects in Mvsidopsis bahia exposed to
Iludioxonil (CGA 173506) Ior 96 hr. under Ilow through conditions.
4 hr. 48 hr. 96 hr.
Average
measured conc.
Mortality
EIIects Mortality
EIIects Mortality
EIIects
Saltwater
control
0
0
10 AN
10 AN
0
0
10 AN
10 AN
0
0
10 AN
10 AN
Solvent control 0
0
10 AN
10 AN
0
0
10 AN
10 AN
0
20
10 AN
8 AN
0.075 0
0
10 AN
10 AN
0
0
10 AN
10 AN
10
0
9 AN
10 AN
0.15 0 1 C, 9 AN 20 8 AN 20 8 AN
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Sun Aug 18 05:04:43 CEST 2013
62
4 hr. 48 hr. 96 hr.
Average
measured conc.
Mortality
EIIects Mortality
EIIects Mortality
EIIects
0 1 C, 9 AN 10 9 AN 10 9 AN
0.24 0
0
10 AN
10 AN
0
30
10 AN
7 AN
0
50
10 AN
2 C, 3 AN
0.37 0
0
10 AN
10 AN
80
80
2 AN
2 AN
100
100
N/A
N/A
0.63 0
0
10 AN
10 AN
100
100
N/A
N/A
100
100
N/A
N/A
AN appeared normal
C lethargic
N/A not applicable
Table 9.2.4-6: Acute toxicity oI Iludioxonil (CGA 173506)

to Mvsidopsis bahia.
Test object Juvenile Mvsidopsis bahia.
LC
50
(C.I.) - 48 hr. 0.30 (0.24 0.37)
LC
50
(C.I.) - 96 hr. 0.27 (0.24 0.37)
LOEC - 96 hr. 0.24
NOEC - 96 hr. 0.15
The study was in accordance with the guideline. It was accepted. The result was that the 96-hr. LC
50
Ior
Iludioxonil Ior mortality oI mysids was 0.27 mg as/L. LOEC and NOEC were 0.24 and 0.15 mg as/L
respectively (Table 9.2.4-6) aIter 96 h.
Report:
Surprenant, D.C. (1990b). (CGA-173506 technical) - Acute toxicity to eastern oysters (Crassostrea virginica)
under Ilow-through conditions.

Syngenta Crop Production unpublished report No. CGA173506/0390.
Springborn Laboratories Inc., Wareham, Massachusetts, USA, unpublished report No. 89-5-2979.
Guidelines:
US EPA FIFRA Guideline No. 72-3, Acute Toxicity Test Ior Estuarine and Marine Aquatic Organisms
GLP: Yes
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The purpose oI the study was to estimate the acute toxicity oI Iludioxonil (CGA 173506) Technical to Eastern
oysters Crassostrea virginica under Ilow-through conditions.
Test substance: Fludioxonil (CGA 173506) Technical, Batch No. P85002, FL No. 881677. Purity 97.5.
Test species: Eastern oyster Crassostrea virginica Irom Aquacultural Research Corporation, Dennis,
Massachusetts.
Test concentrations: Nominal concentrations were 0.084, 0.14, 0.23, 0.39, and 0.650 mg Iludioxonil/L.
Measured mean concentrations were 0.093, 0.150, 0.220, 0.380, and 0.540 mg Iludioxonil/L. The test included
control and solvent control. The selection oI nominal concentration was based on a preliminary study.
Test design: Forty organisms were exposed in duplicate test aquaria (20 per aquarium) in a Ilow-through system
to Iive concentrations oI Iludioxonil, a dilution (seawater) control and a solvent (acetone) control.
Sampling: Visible abnormalities at each 24 hr. interval. AIter 96 hr. oI exposure new shell growth was measured
microscopically to the nearest 0.1 mm.
Statistical analysis: EC
50
-values and 95 conIidence limits were determined by Iitting the untransIormed and
transIormed (probit, log) data to linear curves. William`s test and Bartlett`s test or Kruskal-Wallis` test were
used to determine the lowest concentration at which there was no signiIicant reduction in growth.
Test conditions: The measured temperature was between 18 and 20C, pH ranged Irom 7.7 to 8.0, and the
oxygen concentration was between 6.5 and 7.9 mg/L, salinity ranged Irom 31 to 33
o
/
oo
. Illumination oI the test
area was provided by Iluorescent lights Ior 16 hr. per day. Oysters were Ied algae (I. galbana, T. maculata)
throughout test and acclimatisation.
Results:
During the exposure some undissolved test material crystallised at the tip oI the syringe tubing. No undissolved
test material was observed in the diluter cells or test aquaria. Nominal and measured concentrations are presented
in table 9.2.4-7. Results on oyster shell deposition Irom this 96 hr. experiment with Iludioxonil are presented in
Table 9.2.4-8. EC
50
, NOEC, and LOEC values based on these results are presented in Table 9.2.4-9.
Table 9.2.4-7: Nominal and measured concentrations oI Iludioxonil (CGA 173506) in 96-hr. acute toxicity test
with Eastern oyster Crassostrea virginica under Ilow through conditions.
Measured concentration (mg as/L)
Nominal
(mg as/L)
Day 0
A
Day 0
B
Day 4
A
Day 4
B
Mean SD
0.65 0.51 0.52 0.57 0.58 0.54 0.038
0.39 0.36 0.37 0.42 0.38 0.38 0.026
0.23 0.18 0.21 0.25 0.22 0.22 0.028
0.14 0.13 0.14 0.17 0.18 0.15 0.023
0.084 0.081 0.094 0.099 0.099 0.093 0.0082
Table 9.2.4-8: EIIects oI Iludioxonil (CGA 173506) technical on shell deposition oI Eastern oyster Crassostrea
virginica aIter 96 hr. oI exposure.
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64
Mean and standard deviation
Shell deposition
(mm)
Mean measured concentration
(mg as/L)
Mean SD
Percentage
reduction relative
to pooled control
Solvent control 2.4 1.2
Control 2.5 1.1
Pooled control 2.4 1.1
0.093 2.2 1.1 8.3
0.150 2.1 1.0 13
0.220 1.7 1.0 29
0.380 1.1 0.8 54
0.540 0.6 0.5 75
Table 9.2.4-9: Acute toxicity oI Iludioxonil (CGA 173506) technical to Crassostrea virginica, based on
reduction oI shell growth.
Test object Crassostrea virginica
EC
50
0.37 (0.30 0.44)
LOEC 0.150
NOEC 0.093
The study was accepted. The crystallisation oI test material on the syringe had no eIIect on the results oI the test
(Table 9.2.4-7). The conclusions oI the test Ior EC
50
, LOEC and NOEC oI Iludioxonil Ior shell reduction in
Eastern oyster were values oI 0.37, 0.150, and 0.093 mg as/L respectively (Table 9.2.4-8 and 9.2.4-9).
METABOLITE STUDIES
Report:
Grade, R. (2000a). Acute toxicity test oI CGA 339833 (metabolite oI CGA 173506) to the cladoceran Daphnia
magna Straus in the static system. Syngenta Crop Protection, unpublished report No. CGA339833/0006.
Novartis Crop Protection AG, Basel, Switzerland, unpublished report No. 2003512.
Guidelines:
OECD-Guideline No. 202, Daphnia, Acute Toxicity Test, DraIt July 1999.
GLP: Yes
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The purpose oI the study was to determine the acute toxicity oI Iludioxonil metabolite

CGA 339833 to the
Cladoceran Daphnia magna under Ilow-through conditions.
Test substance: CGA 339833, Batch No. KI6119/6. Purity: 90 + 2 .
Test species: Daphnia magna STRAUS, clone 5. From in house culture. The daphnids used were 24 hr.
Test concentrations: Nominals were 6.25, 12.5, 25, 50 and 100 mg CGA 339833 /L. Controls (blank) were
included. Nominal concentrations were based on range-Iinding test.
Test design: 20 animals per concentration and control (5 animals/replicate). Four replicates Ior the test
concentrations and the control were perIormed. The test was perIormed with Iive test concentrations and one
control.
Sampling: Immobilisation and behaviour at 24 and 48 hr.
Statistical analysis: Normally probit analysis, but necessary because oI absence oI symptoms oI intoxication.
Test conditions: Static. During the test pH varied Irom 8.2 to 8.4, dissolved oxygen varied between 81 and 88
oI saturation, and temperature was 21.5C. during the test. 18 hr. oI light and 8 hr. oI darkness with a 30 min.
transition period. Daphnia were Ied daily.
Results:
The concentration oI CGA 339833 was Iound to be within 99.6 and 102.4 oI the nominal concentrations at test
beginning, and within 99.0 and 103.4 at test termination.
No immobilisation oI daphnids was observed in the controls and all test concentrations.
Table 9.2.4-10: Acute toxicity oI CGA-339833 to Daphnia magna.
Test substance CGA-339833 (mg as/L)
EC
50
(immobilisation) ~ 100
LOEC ~ 100
NOEC 100
The study was accepted, it was in accordance with the guideline. The conclusion was that there was no eIIect oI
the tested metabolite at the highest concentration tested, i.e. 100 mg CGA 339833/L (Table 9.2.4-10).
FORMULATION STUDIES
REFERENCE: Annex KIIIA1 10.2.1
Report:
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Grade R (1994a). Report on the acute toxicity test oI CGA 173506 CGA 219417 62.5 WG (A-9219 B) on
Daphnia (Daphnia magna Strauss 1820). Syngenta Crop Protection, unpublished report No. CGA173506/0471.
Ciba-Geigy Ecotoxicology, Basel, Switzerland. Unpublished report No. 943503.
Guidelines:
OECD-Guideline No. 202.
GLP: Yes
The purpose oI the study was to determine the acute toxicity oI SWITCH 62.5 WG (Iludioxonil cyprodinil) to
the Cladoceran Daphnia magna under semi-static conditions.
Test substance: SWITCH 62.5 WG ( A-9219 B ) Batch No. P311001. Purity: not applicable, Composition
Iludioxonil 25.1, cyprodinil 37.6.
Test species: Daphnia magna. Strain: STRAUS 1820. The daphnids used were 24 hr., Irom Ciba-Geigy Ltd.
Testing Iacility.
Test concentrations: Nominal were 0.01, 0.018, 0.032, 0.058, 0.1, 0.18, and 0.32 mg SWITCH 62.5 WG

/L.
Initial measured were Ior Iludioxonil: 0.007, 0.01, 0.02, 0.06, 0.08, 0.1, and 0.3 mg/L. End measured were
Iludioxonil:

0.007, 0.01, 0.02, lost, 0.06, 0.1, and 0.3 mg/L. Initial measured were Ior cyprodinil: 0.009, 0.02,
0.03, 0.06, 0.08, 0.2, and 0.3 mg/L. End measures were Ior cyprodinil: 0.009, 0.02, 0.03, sample lost, 0.08, 0.2,
and 0.3 mg/L. Control included. Nominal concentrations were based on pre-test.
Test design: Daphnids were exposed to a geometric series oI seven test concentrations, and a negative control.
Four replicate test chambers were maintained in each treatment and control, with 5 daphnids in each test
chamber Ior a total oI 20 daphnids per concentration.
Sampling: Immobilisation at 24 and 48 hr.
Statistical analysis: The EC
50
-values were calculated according to the maximum likelihood method, probit
model.
Test conditions: Semi static. During the test pH varied Irom 7.7 to 7.8, Dissolved oxygen varied between 97 and
100 oI saturation, and temperature was between 19 and 21C. No Ieeding during the test.
Results:
Initial measured concentrations were Ior Iludioxonil: 0.007, 0.01, 0.02, 0.06, 0.08, 0.1, and 0.3 mg /L. End
measured Iludioxonil were:

0.007, 0.01, 0.02, lost, 0.06, 0.1, and 0.3 mg /L. Initial measured were Ior cyprodinil:
0.009, 0.02, 0.03, 0.06, 0.08, 0.2, and 0.3 mg /L. End measured were Ior cyprodinil: 0.009, 0.02, 0.03, lost, 0.08,
0.2, and 0.3 mg/L. The results presented in Table 9.2.4-11 and 12 are based on the nominal concentrations oI
SWITCH 62.5 WG.
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Table 9.2.4-11: Immobilisation oI daphnids exposed to SWITCH 62.5 (A-9219 B; Iludioxonil 25.1, cyprodinil
37.6) in 48 acute toxicity test under semi-static conditions.
24 hr. 48 hr. Nominal
concentration
(mg/L)
Daphnids/test
concentration
Total number Total number
Blank 20 0 0 0 0
0.01 20 0 0 0 0
0.018 20 0 0 0 0
0.032 20 0 0 0 0
0.058 20 0 0 0 0
0.1 20 1 5 1 5
0.18 20 2 10 18 90
0.32 20 18 90 20 100
Table 9.2.4-12: Acute toxicity oI SWITCH 62.5 (A-9219 B; Iludioxonil 25.1, cyprodinil 37.6)

to Daphnia
magna in 48 acute toxicity test under semi-static conditions.
Test substance SWITCH 62.5

(mg /L)*
EC
50
Ior immobilisation aIter 24 hr. (95 C.L.) 0.23 (0.20 0.27)
EC
50
Ior immobilisation aIter 48 hr. (95 C.L.) 0.14 (0.12 0.16)
LOEC 0.1
NOEC 0.058
Based on nominal concentrations
The study was accepted and it Iollowed the guideline. It concluded that the EC
50
Ior the toxicity oI SWITCH
62.5 WG to Daphnia magna was 0.14 mg SWITCH 62.5 WG/L aIter 48 hr., and that the NOEC and the LOEC
were 0.058 and 0.1 mg SWITCH 62.5 WG/L respectively (Table 9.2.4-12). The endpoints Ior the active
ingredient Iludioxonil were EC
50
(48-h) 0.035 mg as/L, LOEC 0.025 mg as/L and NOEC 0.015 (calculated
by RMS based on 25.1 Iludioxonil).
REFERENCE: Annex KIIIA2 10.2.1
Report:
Grade, R. (1999). Acute toxicity test oI CGA 173506, FS 25 (A-8207 I) to the Cladoceran Daphnia magna
STRAUS in the static system, Novartis Crop Protection, Environmental SaIety Ecotoxicology, Report No.
991600. Syngenta Crop Protection Unpublished Report No. CGA173506/5075.
Guidelines: OECD Guideline Ior Testing oI Chemicals, Section 2, No. 202, Daphnia sp. Acute Immobilisation
Test and Reproduction Test.
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68
GLP: Yes
The purpose oI the study was to determine the acute toxicity oI Iludioxonil Iormulation CELEST 025 FS, to the
Cladoceran Daphnia magna.
Test substance: CELEST 025 FS ( A 8207 I ) Batch No. P.812002/l, content oI active ingredient Iludioxonil
25.7 g/L.
Test species: Less than 24 hr. old Daphnia magna STRAUS CLONE 5 Irom Novartis Services
Test concentrations: Nominal A 8207 I were 6.25, 12.5, 25, 50, 100 mg/L.
Test design: Twenty daphnids were exposed to each oI the desired test concentrations Ior 48 hr. under static
conditions in M4-medium. There were Iour replicates Ior the test concentrations and the control. Each replicate
consisted oI 5 species in 100 mL test solution.
Sampling: Immobilisation and behavioural changes aIter 24 and 48 hr. oI exposure.
Statistical analysis: Probit analysis
Test conditions: During the tests varied Irom 7.9 to 8.3, Oxygen varied between 101 and 103 saturation, and
temperature between 21.2 and 21.3C.
Results:
At day 0 the analytical concentrations were between 88 and 94.6 oI the nominal concentrations, at day 2 (48 h)
they varied between 74.2 and 84.8 oI the nominal concentrations except Ior the highest nominal concentration,
where the analytically determined concentration was only 47.9 oI the nominal concentration (Table 9.2.4-13).
Immobilisation occurred in 5 aIter 48 hr. at 25 mg/L nominal, and aIter 24 hr. at 50 mg/L nominal (Table
9.2.4-13).
EC
50
was 46.1 mg/L nominal A 8207 I aIter 48 hr. and 58.3 mg/L nominal A 8207 I aIter 24 hr. (Table 9.2.4-14).
Table 9.2.4-13: Immobilisation oI daphnids exposed to CELEST 025 FS aIter 24 and 48 hr. exposure.
Concentration 24 hr. 48 hr.
Nominal mg/L Measured mg/L Total Total
Blank 0 h 48 h 0 0 0 0
6.25 5.5 5.2 0 0 0 0
12.5 11.8 10.6 0 0 0 0
25 23.6 20.4 0 0 1 5
50 46.5 37.0 9 45 14 70
100 94.6 47.9 17 85 18 90
Table 9.2.4-14

EIIect concentration and NOEC oI CELEST 025 FS in acute toxicity study with Daphnia magna.
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mg CELEST 025 FS /L
nominal concentration
mg CELEST 025 FS /L
mean measured
concentration #
mg Iludioxonil /L
mean measured
concentration *
Test species Daphnia magna
EC
50
24 h (95 C.L.) 58.3 ( 47.8 71.3) - -
EC
50
48 h (95 C.L.) 46.1 (37.5 56.2) 37.93 (31.62-44.78) 0.94
NOEC 24 h 12.5 11.2 0.28
NOEC 48 h 12.5 11.2 0.28
# Value Irom a memo provided by the notiIier November 2004
* Calculated by RMS based on a content oI 2.48 as in the product
The study was in accordance with the guideline and is acceptable. However, the report only gives the endpoints
as nominal concentrations and the notiIier must provide the calculations based on mean measured concentrations
as a report amendment. In this study, the 48 h LC
50
was 37.93 mg CELEST 025 FS /L corresponding to 0.94 mg
Iludioxonil/L (mean measured concentration), NOEC was 11.2 mg CELEST 025 FS /L corresponding to 0.28
mg Iludioxonil/L (mean measured concentration) (Table 9.2.4-14).
B.9.2.5 CHRONIC TOXICITY TO AQUATIC INVERTEBRATES (ANNEX IIA 8.2.5)
REFERENCE: Annex KIIA, 8.2.5
Report:
RuIli, H. (1989a). Report on the Daphnia, reproduction test with CGA 173506 technical, Syngenta Crop
Protection, Unpublished report No. CGA173506/0006. Ciba-Geigy Ltd., Basel, Switzerland. Unpublished report
No. 88 17 36.
Guidelines:
OECD-Guideline No. 202, Phase 2, Paris 1984, modiIied according to the EEC/OECD-ringtest 1985/86.
GLP: Yes
The purpose oI the study was to determine the eIIect oI Iludioxonil on the immobilisation and reproduction oI
Daphnia magna under semi-static conditions.
Test substance: Fludioxonil (CGA 173506) technical, Batch No. 805002, purity 97.5 active ingredient.
Test species: Daphnia magna, Strain: STRAUS 1820.
Test concentrations: Nominal concentrations were 0.00074, 0.0022, 0.0067, 0.020, 0.060 and 0.18 mg
Iludioxonil/L, mean measured concentrations were 0.0004, 0.0009, 0.005, 0.01, 0.03 and 0.1 mg Iludioxonil/L.
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70
The test included control and solvent (acetone) control. The test solutions were renewed each Monday,
Wednesday and Friday.
Test design: Daphnia magna (24h old) were continuously exposed Ior 21 days to a series oI Iive test
concentrations, a solvent control (acetone) and a dilution water control under semi-static conditions. 10 Daphnia
magna per concentration and control/1 daphnia per vessel to insure independence oI the mothers and increase the
strength oI results.
Sampling: Number oI immobilised daphnia, cumulative number oI young/Iemale, cumulative number oI dead
young/Iemale
Statistical analysis: All tests were perIormed using a signiIicance level oI u 0.05.
Test conditions: The measured temperature was 20 + 1C during the test, pH ranged Irom 7.8 to 8.9, the
dissolved oxygen concentration varied between 95 and 133 oI saturation. A photoperiod oI 16 hr. oI light and
8 hr. oI darkness with a light intensity oI 1500 lux. Daily Ieeding oI green algae Scenedesmus subspicatus.
Results:
The measured concentrations (Table 9.2.5-1) were used Ior the calculation oI eIIect concentrations Table 9.2.5-5.
The results on immobilisation and reproduction are presented in Tables 9.2.5-2 to 9.2.5-4. The eIIect
concentrations Ior the toxicity oI Iludioxonil to Daphnia magna are presented in Table 9.2.5-5.
Table 9.2.5-1: Actual concentrations oI Iludioxonil measured at day 0, 7, 14 and 21 expressed as oI nominal
initial concentrations (mg/L) in an exposure oI Daphnia magna.
0 7 14 21 Mean Day
Nominal :
mg Iludioxonil /L
Measured oI nominal
(Day 0 not included in mean)
0.00074 103 57 34 64 51
0.0022 91 36 41 50 42
0.0067 88 94 70 70 78
0.020 90 46 70 48 55
0.060 87 60 65 42 56
0.18 92 58 78 47 61
Table 9.2.5-2: Number oI immobilised parental daphnids Daphnia magna during the 21-day chronic exposure to
Iludioxonil under semi-static conditions.
Number oI immobilised daphnia on day Mean measured
concentration
mg as/L
Day 1 Day 7 Day 14 Day 21
Control 0 0 0 1
Solvent cont. 0 0 0 1
0.0004 0 0 0 5
0.0009 0 0 0 1
0.005 0 0 0 2
0.01 0 0 0 0
0.03 0 0 0 5
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71
Number oI immobilised daphnia on day Mean measured
concentration
mg as/L
0.1 0 0 0 4
Table 9.2.5-3: Cumulative average number oI young/Iemale at day 7, 14, 21 Ior Iludioxonil (mg/L) exposed
Daphnia magna in 21 day chronic exposure semi-static test.
(mg/L)
Cumulative number oI young/Iemale
(dead and alive)
Cumulative number oI dead young/Iemale
Day 7 14 21 7 14 21
Control 0 37 62 0 5 7
Solvent cont. 0 32 54 0 5 7
0.0004 0 25 53 0 13 17
0.0009 0 34 70 0 5 13
0.005 0 34 68 0 9 17
0.01 0 27 59 0 11 20
0.03 0 26 52 0 10 12
0.1 0 5 7 0 3 3
Table 9.2.5-4: Length oI time Ior appearance oI Iirst brood Ior Iludioxonil (mg /L) exposed Daphnia magna in
21 day chronic exposure semi-static test.
Number oI daphnia with Iirst brood appearing on day
Measured
concentration
(mg/L)
9 12 14 ~14
Control 9 1 0 0
Solvent cont. 7 3 0 0
0.0004 7 3 0 0
0.0009 8 2 0 0
0.005 6 4 0 0
0.01 1 9 0 0
0.03 0 10 0 0
0.1 0 3 1 6*
* 3 oI 10 dapnia died beIore producing any young
Table 9.2.5-5: Chronic toxicity oI Iludioxonil

to Daphnia magna based on mean measured conc.
Test duration 21 d.
Immobility
EC
50
~ 0.1
LOEC ~ 0.1
NOEC 0.1
Reproduction
EC
50
0.03 0.1
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72
Test duration 21 d.
LOEC 0.1
NOEC (Time Ior appearance oI Iirst brood) 0.005
NOEC (Fraction oI dead young) 0.005
NOEC (Total cumul. No. oI young) 0.03
The test Iollowed the guideline. The test was acceptable. For the Daphnia magna 21-day reproduction toxicity
test with Iludioxonil, the Iraction oI dead young and the time Ior appearance oI Iirst brood were the most
sensitive biological parameters. NOEC Ior reproductive toxicity oI Iludioxonil on Daphnia magna was 0.005 mg
as/L (9.2.5-5).
REFERENCE: Annex KIIA, 8.2.5
Report:
Putt A.E. (1991). (CGA-173506 technical) Chronic toxicity to daphnids (Daphnia magna) under Ilow-through
conditions. Syngenta Crop Protection, Unpublished report No. CGA173506/0179. Springborn Laboratories Inc.,
Wareham, Massachusetts, United States, Unpublished report No. 91-2-3672.
Guidelines:
EPA Pesticide assessment Guidelines, E, WildliIe and Aquatic Organisms, 72-4.
GLP: Yes
The purpose oI the study was to estimate the chronic toxicity oI Iludioxonil (CGA 173506) technical to daphnids
Daphnia magna under Ilow-through conditions.
Test substance: Fludioxonil (CGA 173506) technical, Batch No. P805002, FL No. 881677, purity 97.5 active
ingredient.
Test species: The daphnids (Daphnia magna) used in this toxicity test were obtained Irom populations cultured
at Springborn Laboratories Inc.
Test concentrations: Nominal concentrations were 0.020, 0.040, 0.080, 0.160, 0.320 mg Iludioxonil/L, mean
measured concentrations were 0.019, 0.034, 0.062, 0.086, 0.190 mg Iludioxonil/L. The test included control and
solvent control.
Test design: Daphnia magna (24h old) were continuously exposed Ior 21 days to a series oI Iive test
concentrations, a solvent control (acetone) and a dilution water control under Ilow-through conditions. Four
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73
replicate test chambers were maintained in each treatment and control group with 10 Daphnia magna in each test
chamber.
Sampling: Adult survival was recorded on days 1, 2, 4, 7, 9, 11, 14, 16, 18 and 21. Measurements oI oIIspring
production were recorded on days 9, 11, 14, 16, 18 and 21. At each observation interval, the oIIspring was
removed, counted and discarded. The total body length and dry weight oI all surviving adult organisms were
measured at test termination.
Statistical analysis: Student`s T-Test Ior control-solvent control comparison. Chi Square compared observed
distribution with normal distribution. Bartlett`s test Ior homogeneity oI variance. William`s test, Dunnett`s test
and Kruskal-Wallis test Ior comparison oI test concentration with solvent control daphnids.
Test conditions: The measured temperature was 20C during the test, pH ranged Irom 7.6 to 8.1, and the oxygen
concentration was between 7.8 and 8.7 mg/L. A photoperiod oI 16 hr. oI light and 8 hr. oI darkness with a light
intensity oI 24-50 Ioot-candles at the test solution surIace was maintained throughout the test period.
Results:
Reproduction results (based on the mean measured concentrations) are presented in the Tables 9.2.5-6 to 9.2.5-
7. Comparisons oI nominal and measured concentrations oI Iludioxonil are presented in Table 9.2.5-8. At the
three highest test concentrations some material was not dissolved. EIIect concentrations Ior the chronic toxicity
oI Iludioxonil to Daphnia magna are presented in Table 9.2.5-9.
Table 9.2.5-6: Mean percentage survival Ior parental daphnids Daphnia magna during the 21-day chronic
exposure to Iludioxonil.
Survival Mean measured
concentration (mg as/L)
0.190 98
b
88
b
85
b
80
ab
0.086 95
b
95
b
95
b
95
b
0.062 100 100
b
100
b
100
b
0.034 100
b
100 100
b
98
b
0.019 100 100 100 100
Solvent cont. 100 100 100 100
Control 100 100 100
b
93
a
SigniIicantly diIIerent (p 0.05) as compared to the solvent control data.
b Daphnia exhibiting abnormal signs.
Table 9.2.5-7: Cumulative number oI oIIspring as a Iunction oI time Ior Iludioxonil (mg/L) exposed Daphnia
magna in 21 day chronic exposure Ilow-through test.
Total number oI oIIspring/Iemale
Mean (SD
a
)
Day Control Solvent
control
0.019
(mg /L)
0.034
(mg /L)
0.062
(mg /L)
0.086
(mg /L)
0.190
b
(mg /L)
9
4(4) 1(1) 0(0) 0(0) 0(0) 0(0) 0(0)
11
28(20) 27(9) 4(4) 0(0) 0(0) 0(0) 0(0)
14
70(27) 71(11) 44(10) 27(9)
c
7(4) 0(0) 0(0)
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Total number oI oIIspring/Iemale
Mean (SD
a
)
16
75(31) 73(13) 52(13) 31(8) 14(9) 1(1) 0(0)
18
122(39)
d
98(12) 85(28) 31(8) 15(9) 1(1) 0(0)
21
170(48) 111(13)
I
103(14) 31(8)
e
15(9)
e
1(1)
e
0(0)
a
SD Standard Deviation
b
Due to an adverse eIIect on organism survival, reproduction data Ior this treatment level was not statistically analysed Ior
treatment eIIects
c
Two oI the oIIspring were immobilised
d
One oI the oIIspring was immobilised
e
SigniIicantly diIIerent (p 0.05) as compared to the solvent control data.
I
Several oI the oIIspring were immobilised.
Table 9.2.5-8: Actual concentrations oI Iludioxonil measured at day 0, 7, 14 and 21 expressed as oI nominal
initial concentrations (mg/L) in an exposure oI Daphnia magna.
Measured concentration ( oI nominal) Nominal concentration
mg Iludioxonil /L
0.320 47 39 47 109
0.160 32 61 33 88
0.080 53 85 61 112
0.040 73 91 83 100
0.020 88 93 93 105
Table 9.2.5-9: Chronic toxicity oI Iludioxonil to Daphnia magna.
Test substance Fludioxonil
(mg as/L)*
Test duration 21 d.
Immobility Reproduction
EC
50
~ 0.190
LOEC 0.034
NOEC 0.019
*Based on mean measured concentrations
Results are very poorly presented in the report. It should be noted that the mean measured concentration
probably is a result oI material not dissolved, being dissolved on the sampling day 21 at the termination oI the
study, thereIore the calculated mean measured concentration is likely to be overestimated Ior the three highest
test concentrations. However the NOEC could be considered valid because it was obtained at a low
concentration. The NOEC Ior chronic toxicity oI Iludioxonil to Daphnia magna was 0.019 mg as/L. The EC
50
ought to be adjusted to measured concentrations (e.g. Ior day 0, 7 and 14, which is 0.1417 mg as/L). The study
Iollowed the guideline. The study was acceptable Ior the derivation oI the NOEC, which is the relevant endpoint
Ior the risk assessment.
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