3,5-bis(2,4-dimethylcyclohex-3-en-1-yl)polyheterocycle

Toxicity to soil macroorganisms except arthropods

Administrative data

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-08-10 to 2019-04-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Sampling and analysis

Analytical monitoring:

no

Test substrate

Vehicle:

no

Details on preparation and application of test substrate:

The artificial soil was prepared as follows:
a) 10 per cent sphagnum peat (pH 5.62, the water content measured before use was 1.78%)
b) 20 per cent kaolin clay (kaolinite content was greater than 30%);
c) 70 per cent industrial sand (fine sand should be dominant with more than 50 per cent of the particles between 50 and 200 microns);
According to above proportions, prepare 6 groups of 2500 g artificial soil for he test (50 g industrial sand was deducted from amount of industrial sand to use for mixing with test item). The dry constituents are blended in the correct proportions and mixed thoroughly in blender. The moisture content was adjusted to about 20% of the dry weight with distilled water (500 ml). After test item application, the moisture content was adjusted to about 30-35% (the mixture should be moist but not so wet that water appears when the artificial soil is compressed). pH of artificial soil was determined before the test.
0.2507, 0.4520, 0.8020, 1.4056, 2.5060 g of test item were weighed and mixed with 50 g industrial sand respectively. The treated sand was mixed thoroughly with proper artificial soil (dry weight was 2450 g), and distilled water (250 ml) was added to adjust the moisture content to about 30% of dry weight and mixed to be homogeneous. For each treated soil, four 750 g of the wet test medium were weighed and put into four labelled test containers with glass plate.
A concurrent control group with four replicates maintained under identical conditions only without test item application was performed in parallel. 50 g industrial sand was applied to properly pre-moistened artificial soil ( dry weight was 2450 g), then distilled water (250 ml) was added to adjust the moisture content to about 30% of dry weight and mixed to be homogeneous.

Test organisms

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

The test earthworm (Eisenia foetida) was obtained from Chenggong Earthworm Raising Farm in Dongli District, Tianjin. The domestication conditions of earthworms were 20±2° C, 80±15% relative humidity and continuous light.
The adult (at least two months old with clitellum) worms with an individual weight of 0.3023~0.5604 g were used in the test.

Study design

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test conditions

Test temperature:

18,7-20,3 °C

pH:

5.51-5.91

Moisture:

29.8-29.9%

Details on test conditions:

Test concentrations included I 00, 180, 320, 560 and 1000 mg/kg dry soil weight. A blank control group was conducted in parallel. Four replicates were used for the test groups and control group.
For each test container, 10 earthworms (which had been conditioned in an artificial soil, washed, dried on filter paper and weighed before use) were placed on the surface of test medium. The containers were covered with a glass plate to prevent the earthworms from escaping and the test medium from drying. The containers were kept under the test conditions for 14 days.
he mortality was assessed by emptying test medium onto a plate, sorting worms from the medium and testing their reaction to a mechanical stimulus at the front end at the 7th and 14th day. Any behavioural signs was recorded. After the 7-day assessment worms and medium was replaced in the test containers.
At the start and the end of the test, the individual weights of all live worms in each group were measured and recorded. The worms were washed prior to weighing (with distilled water) and the excess water was removed by placing the wonns briefly on filter paper.
At the beginning and the end of the test, soil moisture content was determined at the test groups and control medium. 1 replicate at the start and 4 replicates at the end. The moisture content was determined by drying a small sample at 105°C for 3 hours and re-weighing.
At the beginning of the test, soil pH was measured. I replicate was measured in test group and control group. The method is as followed: wet soil was dried at room temperature for at least 21 h. A suspension of the soil ( containing 5 grams of soil) was then made up in five times its volume of a 0.01 M solution of analytical grade calcium chloride (CaCl2). The suspension was then shaken thoroughly for six minutes and then left to settle for at 2 hours. The pH of the liquid phase is then measured using a pH-meter.

Nominal and measured concentrations:

180, 320, 560 and 1000 mg/kg dry soil weight

Reference substance (positive control):

yes

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

207 mg/kg soil dw

Basis for effect:

mortality

Effect concentrations 2

Duration:

7 d

Dose descriptor:

LC50

Effect conc.:

341.6 mg/kg soil dw

Basis for effect:

mortality

Details on results:

During the exposure period, there was no mortality or abnormal signs observed at control group and test group of 100 mg/kg dry soil weight; At test groups of 180, 320, 560 and 1000 mg/kg dry soil weight, there were flexible body, loose and disconnect canker observed, and earthworms even dead, and the number of deaths increased with increasing concentrations, all earthworms dead in 7th day of 1000 mg/kg dry soil weight test group.

Results with reference substance (positive control):

The 14-day LC50 of the reference substance to earthworm is 21.1 mg a.i./kg dry soil weight, with 95% confidence limits of 17.7-25.1 mg a.i./kg dry soil weight, which met the requirement of 20-80 mg a.i./kg dry soil weight in the guideline.

Reported statistics and error estimates:

Trimmed spearman-karber method (TSK,Version 1.5) was used to evaluate the data, LC50 and 95% confidence limit.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

According to the results, the acute toxicity of Sa 57 to the earthworm (Eisenia foetida) was:
LC50 ( 7 d): 341.6 mg/kg dry soil weight > 95% confidence limit were 300.1-388.8 mg/kg dry soil weight;
LC50 (14 d): 207.0 mg/kg dry soil weight >95% confidence limit were 184.0-232.8 mg/kg dry soil weight

Executive summary:

This study was performed to assess the acute toxicity of Sa 57 to the earthworm (Eiseniajoetida).
Based on the results of the preliminary test, the definitive study was conducted at concentrations of 100, 180, 320, 560 and 1000 mg/kg dry soil weight. A concurrent control group was conducted in parallel. For the test and control group, 4 replicates were used with 10 worms in each replicate.
Artificial soil was used in this study. The test duration was 14 days, assessment of mortality and abnormal signs was conducted at 7 and 14 day of test.
During the exposure period, there was no mortality or abnormal signs observed at control group and test group of 100 mg/kg dry soil weight; At test groups of 180, 320, 560 and 1000 mg/kg dry soil weight, there were flexible body, loose and disconnect canker observed, and earthworms even dead, and the number of deaths increased with increasing concentrations, all earthworms dead in 7th day of 1000 mg/kg dry soil weight test group.

According to the results, the acute toxicity of Sa 57 to the earthworm (Eisenia foetida) was:

LC50 ( 7 d): 341.6 mg/kg dry soil weight > 95% confidence limit were 300.1-388.8 mg/kg dry soil weight;

LC50 (14 d): 207.0 mg/kg dry soil weight >95% confidence limit were 184.0-232.8 mg/kg dry soil weight