(6R-trans)-7-amino-8-oxo-3-[[(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-1,2,4-triazin-3-yl)thio]methyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Version / remarks:

1996

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

For determination of the test item concentrations samples were taken from the test concentration and the control at the start and at the end of each water renewal period.

Vehicle:

no

Details on test solutions:

The test solution used in the test was prepared by mechanical dispersion without using of any solubilising agent. The test solution was prepared by dissolving an amount of 0.03 g test item in 300 mL dilution water (ISO medium) resulting a nominal concentration of 100 mg/L. This solution was ultrasonicated for approx. 15 min and then agitated by orbital shaker for about 1 hour to obtain clear solution. The test solution was prepared just before introduction of test animals (start of each renewal period).

Test organisms (species):

Daphnia magna

Details on test organisms:

Species and strain: Daphnia magna (Straus)
Origin: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3. Hungary
Breeding: The Daphnia are bred in the Laboratory of TOXI-COOP ZRT.
Number of animals: Twenty animals in both groups (concentration and untreated control), divided into 4 concurrent batches (5 animals per batch).
Age of animals: Less than 24 h old at the beginning of the test.
Sex: Female
Animal health: Apparently healthy animals were used in this test with a known history
Acclimatization: Test animals were bred under similar (or the same) conditions as that used during the exposure period (holding water, temperature, background colour etc.), therefore additional acclimatisation before the test was not necessary. Brood daphnids were maintained in dilution water at the test temperature for at least 48 hours prior to the start of the test.
Food and Feeding: The Daphnia were fed with concentrated algal suspension of Raphidocelis subcapitata during the holding. Test animals were not fed during the exposure.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

According to Guidelines.

Post exposure observation period:

No.

Hardness:

The reconstituted water (ISO medium) had an approximate theoretical total hardness of 249 mg/L (as CaCO3).

Test temperature:

The test temperature was in the range of 19.9 – 20.4°C measured in the test vessels during the test. The additionally measured ambient temperature within the climate chamber was between 19.8 – 20.6°C.

pH:

The pH of the test solution was not adjusted and not varied by more than 1.5 units in any one test group. The pH was in the range of 7.07 – 8.00 during the test.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 7.62 – 8.04 mg/L during the test.

Salinity:

No.

Conductivity:

Not reported.

Nominal and measured concentrations:

0 - 100 mg/L.
No toxic effects were observed during the preliminary test therefore, a single concentration of 100 mg/L (limit concentration) and a concurrent control were included in the main test.
The analytically measured test item concentrations remained within ± 20 % of the nominal during the test. Therefore, the biological endpoints are based on the nominal test item concentration

Details on test conditions:

Test type: Semi-static test. Based on the results obtained during analytical method validation the test item is not stable for the duration of 48 hours in ISO Medium, therefore a semi-static test was performed with daily water renewal periods.

Reconstituted water (ISO medium, according to OECD 202) was used as dilution water in the experiment.

The test vessels were kept during the test in a climate chamber under controlled conditions. The temperature, dissolved oxygen concentration and pH were measured in the test item treated and the control group at the start (before test solutions had been distributed into the test vessels) and in each test vessel at the end of the test. Additionally, the ambient temperature was measured continuously using a min/max thermometer in the climate chamber.

Light-Dark Cycle: The light-dark cycle during the test was 16 hours light (artificial illumination) and 8 hours darkness.

In order to select appropriate test concentrations for use in the definitive test, a non-GLP preliminary range-finding test was conducted to determine the approximate toxicity of the test item. No toxic effects were observed during the preliminary test therefore, a single concentration of 100 mg/L (limit concentration) and a concurrent control were included in the main test.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate.

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

There was no significant immobilisation observed in twenty daphnids exposed to each test item treated and control group after 48 hours of exposure. The immobilisation rate was 10 % in the control and 5 % in the 100 mg/L concentration. These immobilisations were within the normal range.
Further no abnormal behaviour or appearance of test animals was noted.

Results with reference substance (positive control):

The date of the last study (Study Code: 392-202-2533) with reference item Potassium dichromate was: 07 – 08 March 2017.
The 24-h EC50: 1.41 mg/L, (95 % confidence limits: 1.19 – 1.68 mg/L).

Validity of the Study

• The immobilisation rate did not exceed 10 % either in the control or in the test item treated group.

• The dissolved oxygen concentration was greater 3 mg/L in each test vessel at the end of the test.

All validity criteria were within acceptable limits and therefore the study considered valid.

The concentration of the test item was analytically determined at the start and at the end of each water renewal period. In the untreated control group the test item was not detected. The measured concentrations of 7-ACT varied between 96.5 and 110 % of the nominal during the test period.

Validity criteria fulfilled:

yes

Conclusions:

The test item had no toxic effect at 100 mg/L concentration on Daphnia magna; the EC50 and the LOEC are higher than 100 mg/L.

Executive summary:

A semi-static test with Daphnia magna was performed according to OECD guideline 202. The test item had no toxic effect at 100 mg/L concentration (i.e. limit test concentration) on Daphnia magna. The NOEC48h is 100 mg/L. The EC10, EC20, EC50 results and the LOEC are higher than 100 mg/L.

Description of key information

The test item had no toxic effect at 100 mg/L concentration (i.e. limit test concentration) on Daphnia magna. The NOEC48h is 100 mg/L. The EC10, EC20, EC50 results and the LOEC are higher than 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The EC50 is not 100 mg/L, as one is forced to enter in the field above, but the EC50 is >100 mg/L.