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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Reference
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 Apr - 09 Jun 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
GLP compliance:
yes (incl. certificate)
Remarks:
Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): As the test item is insoluble in water and easily evaporating organic solvents, the quantity of test item required to obtain the desired concentrations was applied to quartz sand by grinding. The spiked quartz sand was distributed on the soil and mixed thoroughly. The test substrate was prorated to each test container.
- Controls: yes
Test organisms (species):
Eisenia andrei
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Source: Regenwurmfarm Tacke, Borken, Germany
- Age at test initiation (mean and range, SD): 2 - 12 months
- Weight at test initiation (mean and range, SD): 250 - 600 mg

ACCLIMATION
- Acclimation period:6 days
- Acclimation conditions (same as test or not): The worms were conditioned in the artificial soil before use. The same food which was also used in the test was given in a sufficient amount.
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Test temperature:
18 - 22°C
pH:
test start: 6.2 - 6.5
test end: 5.9 - 6.2
Details on test conditions:
TEST SYSTEM
- Test container (material, size): plastic containers (area of 17x12 cm, height of 6 cm); containers were covered with perforated transparent plastic caps
- Amount of soil or substrate: 500 - 650 g dry mass of artificial soil
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8
- other: One day after addition of the worms, an amount of 5 g air dried finely ground cow manure per test container was spread on the soil surface and moisten with water. Once a week, during the first four weeks, the adult earthworms were fed according to their food consumption.

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): Sphagnum peat, air-dried, finely ground 10 %, Kaolinite, air-dried 20 %, Industrial quartz sand, air-dried 70 %
- Pretreatment of soil: The test substrate was wetted with deionised water to a water content of between 40 % and 60 % by mass of the total water-holding capacity. The pH of the substrate was adjusted to 6.0 ± 0.5 (1 mol/L KCl) by adding pulverized calcium carbonate (CaCO3).

OTHER TEST CONDITIONS
- Photoperiod: light/dark cycle 16 h : 8 h
- Light intensity: 732 - 764 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality and weight change after 28 days, offspring number after 56 days
Nominal and measured concentrations:
Nominal: control, 62.5, 125, 250, 500 and 1000 mg TI/kg dry mass soil
Reference substance (positive control):
yes
Remarks:
carbendazim
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
500 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: weight change
Duration:
28 d
Dose descriptor:
EC0
Effect conc.:
750 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: weight change
Details on results:
- Mortality at end of exposure period: ≤ 5%
- Total mass of adults at beginning of test: see "Any other information on results"
- No. of offspring produced: see "Any other information on results"
Results with reference substance (positive control):
- Results with reference substance valid?
- Relevant effect levels: EC10 = 0.9 mg/kg for weight change; EC10 = 2.0 mg/kg for reproduction (results for reproduction are in agreement with the range of 1 - 5 mg carbendazim per kg (OECD 222); weight change and mortality are in agreement with experience in test facikity)
- Other:

Living earthworms after 28 days. Single values of the parallel test pots [Individuals].

TI = Test item; DM = Dry mass; Concentrations given as nominal values.

 

replicate

Control

62.5 mg TI
per kg DM

125 mg TI
per kg DM

250 mg TI
per kg DM

500 mg TI
per kg DM

1000 mg TI
per kg DM

Living earthworms [Ind.]

1

9

10

9

10

10

9

2

10

9

10

10

9

10

3

10

10

10

9

10

10

4

9

10

9

10

10

9

5

10

 

 

 

 

 

6

10

 

 

 

 

 

7

10

 

 

 

 

 

8

8

 

 

 

 

 

 

Biomass of the earthworm batches at test start and after 28 days.Single values of the parallel test pots [g FM].

         TI = Test item; FM = Fresh mass; DM = Dry mass; Concentrations given as nominal values.

 

replicate

Control

62.5 mg TI
per kg DM

125 mg TI
per kg DM

250 mg TI
per kg DM

500 mg TI
per kg DM

1000 mg TI
per kg DM

Test start

1

3.588

3.062

3.131

3.187

3.408

3.752

2

3.302

2.977

3.554

3.295

2.962

3.313

3

3.064

3.257

3.378

3.069

3.002

3.327

4

2.965

3.408

3.272

3.080

3.076

3.426

5

3.441

 

 

 

 

 

6

3.364

 

 

 

 

 

7

3.409

 

 

 

 

 

8

3.524

 

 

 

 

 

28 d

1

4.885

4.917

4.696

4.911

5.098

3.898

2

4.995

4.063

5.237

5.247

4.227

4.364

3

4.664

4.749

5.251

4.325

4.839

4.684

4

4.211

4.966

4.652

4.801

4.692

4.152

5

5.223

 

 

 

 

 

6

5.061

 

 

 

 

 

7

5.267

 

 

 

 

 

8

4.175

 

 

 

 

 

 

Weight change at test end. Weight change compared to the start value [%] and compared to the control [%].

TI = Test item; DM = Dry mass; *: significant when compared with control (Williams test, one-sided); Concentrations given as nominal values.

Test species

Control

62.5 mg TI
per kg DM

125 mg TI
per kg DM

250 mg TI
per kg DM

500 mg TI
per kg DM

1000 mg TI
per kg DM

compared to start value [%]

45

47

49

53

52

24

compared to control [%]

 

6

9

18

15

-45 *

 

Reproduction. Single values of the parallel test pots [Individuals]and mean values at the end of the study (56 d) [Individuals].

TI = Test item; DM = Dry mass; CV = Coefficient of variation; Concentrations given as nominal values.

 

Replicate

Control

62.5 mg TI
per kg DM

125 mg TI
per kg DM

250 mg TI
per kg DM

500 mg TI
per kg DM

1000 mg TI
per kg DM

Juveniles [Ind.]

1

120

142

79

120

112

94

2

116

140

87

112

89

57

3

127

143

143

48

134

126

4

48

129

82

83

114

111

5

97

 

6

69

7

118

8

129

Mean Juveniles [Ind]
(CV)

 

103
(28.8)

139
(4.7)

98
(31.0)

91
(36.0)

112
(16.4)

97
(30.6)

 

 

Validity criteria fulfilled:
yes

Description of key information

Key value for chemical safety assessment

Additional information

One experimental study was conducted investigating the effects of Boron orthophosphate (CAS 13308-51-5) on mortality, body weight and reproduction of Eisenia andrei. Until now preliminary results are available.

The test was performed according to OECD 222 with nominal concentrations of 62.5, 125, 250, 500 and 1000 mg/kg. After 4 weeks mortality of adults was ≤ 5%, resulting in a NOEC ≥ 1000 mg/kg soil dw. For weight change a NOEC (4 wks) of 500 mg/kg soil dw was calculated. After an additional 4 weeks the influence of the test substance on reproductive output of parent animals was recorded. No effects on the number of juveniles were found resulting in a NOEC (8 wks) ≥ 1000 mg/kg soil dw. Thus, the relevant endpoints mortality and reproduction did not result in any effects up to a nominal concentration of 1000 mg/kg soil dw.