Ceratonia siliqua, ext.

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 October 2017 - 15 November 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

liquid

Details on test material:

- Physical appearance: Dark brown to black liquid
- Storage of test material: At room temperature

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

For the final test, samples for analysis were taken from all test concentrations and the control according to the schedule below:
Frequency: at t=0 and t=96
Volume: 2.0 mL
Storage: Samples were stored in a freezer (≤-15°C) until analysis

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- The test item was completely soluble in test medium at the test concentrations initially prepared. No correction factor was applied.
- Method: Direct application to the test medium. Preparation of test solutions started with the highest concentration of 100 mg/L applying a 1¼ to 2 hour period of magnetic stirring to accelerate dissolution of the test item in medium. Lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium.
- Evidence of undissolved material: All test solutions, besides the highest concentration, were clear and colorless at the end of the preparation procedure. The highest concentration was slightly yellow.
- Controls: Test medium without test item or other additives.

Test organisms

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: carp
- Strain: Linnaeus, 1758
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands
- Age at study initiation: not indicated, F1 from a single parent-pair
- Length at study initiation (final test): 2.7 ± 0.3 cm
- Weight at study initiation (final test): 0.24 ± 0.08 g
- Method of breeding: in UV-treated water

ACCLIMATION
- Acclimation period: at least 12 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: daily with pelleted fish food (Essence (300-500 um), Coppens International bv, Helmond, The Netherlands)
- Health during acclimation: mortality during the seven days prior to the start of the test was less than 5%

FEEDING DURING TEST: no

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

180 mg CaCO3/L

Test temperature:

Throughout the test: 21-22°C (control and test solution)

pH:

Throughout the test: 7.6-8.2 (control) and 7.5-8.1 (test solution)

Dissolved oxygen:

Throughout the test: 7.2-10 mg O2/L (control) and 6.2-9.4 mg O2/mL (test solution)

Nominal and measured concentrations:

Nominal concentration: 100 mg/L
Measured concentrations: at t=0 74.8 mg/L, at t=96 80.9 mg/L
The measured test concentration at the end of the exposure period was 108% of the measured test concentration at the start of the exposure period.

Details on test conditions:

TEST SYSTEM
- Test vessel: 6.5 litres, all-glass, fill volume: 5.0 litres
- Aeration: introduced after 48 hours of exposure
- No. of organisms per vessel: 7
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- Biomass loading rate: 0.34 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Test medium used: adjusted ISO medium according to OECD guideline 203
- Source/preparation of dilution water: tap-water purified by reverse osmosis
- Culture medium different from test medium: no
- Intervals of water quality measurement: dissolved oxygen content, pH and temperature: daily in all vessels with surviving fish, beginning at day 0.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours, daily

EFFECT PARAMETERS MEASURED: mortality at t= 2 1/4, 24, 48, 72 and 96 hours

RANGE FINDING STUDY
- Test concentrations: 0.10, 1.0, 10.0 and 100 mg/L
- Test conditions different from test: 3 fish were exposed to each test concentration and aeration was introduced after 72 hours of exposure.
- Results used to determine the conditions for the definitive study: yes, since no mortality or clinical effects were observed at any of the test concentrations during the test period, the LC50 was expected to exceed 100 mg/L. Based on these results, a limit test was performed as the main study.

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol (February 2017)

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 75 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality (fish)

Details on results:

- Observations in the range-finding test: No mortality occurred. After 24 hours, feaces were observed in the two highest test concentrations. After 48 hours the test solutions of the two highest test concentrations were slightly hazy. However, after 48 hours no feaces and after 72 hours no haziness were observed. Analysis of the test samples (100 mg/L) showed that the test concentration remained stable during the exposure period.

- Mortality or abnormal observations during the final test: no
- The test solution at the end of the exposure period was slightly hazy at the end of the test.
- Analysis of the samples taken at the start of the final test showed a measured concentration of 75 mg/L at the nominal concentrations of 100 mg/L. After 96 hours of exposure the test item concentrations were at the level of initial (108%). Therefore the effect parameter was expressed in terms of the initial measured concentration at t=0 h.
- The pH, dissolved oxygen concentrations and temperature remained within the ranges prescribed by the study plan.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Test concentrations: 0.10, 0.22 and 0.46 mg/L
- Mortality: all carp exposed to a concentration of 0.46 mg/L died within 24 hours, while no mortality occurred at 0.10 and 0.22 mg/L.
- LC50: 0.32 mg/L (95 % confidence interval between 0.22 and 0.46 mg/L)
- Other: since the results with the reference substance were within the historical data range, the carp used in this study are considered to be of anticipated sensitivy.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

see 'overall remarks'

Conclusions:

Based on the results of an acute toxicty study in fish, according to OECD guideline 203 and GLP principles, the LC50 of Carbon Bean Extract exceeded the highest test concentration, i.e. >75 mg/L (initial measured concentration).