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EC number: 226-789-3
CAS number: 5468-75-7
Mortality of Adult Earthworms in
[%] after 28 Days of Exposure
[mg test item/kg soil dry weight]
Earthworm Behaviour and
Pathological Symptoms after 28 Days of Exposure
A): no obersvaton of symptoms or behavioural changes; H) dead earthworm;
D) spontaneous segmentation and separation
Body Weight Changes of the Adult
Application rate[mg test item/ kg soil dry weight]
Mean body weights per replicate
Mean body weight change of earthworms
Mean ± SD
0.23 ± 0.04
47.8 ± 7.81
0.25 ± 0.04
53.0 ± 8.34
52.3 ± 8.77
Rep:= Replicate; Sign. = Significance
Reproduction Rate (Number of
Juveniles after 8 Weeks)
Application rate[mg test item/kg soil dry weight]
Number of juveniles
of pooled control
Significance compared to pooled control
304 ± 41.9
291 ± 44.9
343 ± 23.8
of the test item
on mortality, biomass and the reproductive potential of the earthworm
species Eisenia fetida (Savigny) were determined according to the
guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994) from November
19th, 2009 to January 16th, 2010 at
31157 Sarstedt, Germany. The study was conducted under static conditions
over 8 weeks with a limit test item concentration of 1000 mg test
item/kg soil dry weight, which was mixed into artificial soil containing
10 % peat. A control using untreated artificial soil and a solvent
control with the solvent mixed into the artificial soil were set up. 80
test organisms, with an individual weight of 0.30 to 0.60 g, were
divided into 8 replicate groups with 10 test organisms per replicate.
28 days no evident mortality as well as no pathological symptoms or
changes in the behaviour of adult earthworms were seen in the controls
and at the limit test item concentration of 1000 mg test item/kg soil
dry weight. The earthworm biomass
increased in the controls and at the limit test item concentration of
1000 mg test item/kg soil dry weight
over 28 days without any statistically significant differences between
pooled control and limit test concentration.
further four weeks the reproduction rate (average number of juveniles)
was 297 in the pooled control
and 343 at the limit test item concentration.
Compared to the pooled control, the reproduction rate was not
statistically significantly reduced, but increased at 1000 mg test
item/kg soil dry weight.
the NOEC of the test item concerning earthworm mortality, biomass and
reduction of reproduction was determined to be at 1000
mg test item/kg soil dry weight.
validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in
the Tested Dose
Mortality of adult earthworms
Reduction of body weight
Reduction of reproduction rate
(Number of juveniles)
Summary of all effects
NOEC and LOEC
for Reproduction, Mortality and Body Weight
NOECreproduction, mortality, body weight
LOECreproduction, mortality, body weight
Under the conditions of this study, the test item did not induce evident
mortality or pathological symptoms of adult earthworms after an exposure
to the test item in a concentration of 1000 mg/kg soil dry weight for 28
days. Also, the biomass increase of the adult earthworms after 28 days
was not statistically significantly reduced compared to the pooled
control at the test item concentration of 1000 mg test item/kg soil dry
weight. After eight weeks of exposure the earthworm reproduction was not
statistically significantly reduced at the limit test item concentration
of 1000 mg test item/ kg soil dry weight compared to the pooled control.
The results from the two long term toxicity tests on Eisenia fetida with
Pigments Yellow 12 and its structural analogue, Pigment Yellow 83,
conclusively show absence of toxicity to earth worms, neither in regard
to reproduction, nor in regard to mortality and body weight.
As long term toxicity tests towards soil macro-organisms except
arthropods (earth worm Eisenia fetida) for two different pigments of the
Diarylide Yellow group had been conducted with test substance
concentrations far above the solubility limit in water, from acute tests
no additional information could be expected. Therefore, no acute toxicity
tests were performed.
Judging from these long-term toxicity tests with two different pigments,
the Diarylide Yellow Pigments are to be regarded as nontoxic to soil
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