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Ecotoxicological information

Long-term toxicity to fish

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Reference
Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
The dissolved oxygen concentrations were not maintained at 70% or more of the air saturation value throughout the test, this was a consequence of testing a gas in air-tight closed vessels. However, no negative effects were observed.
Qualifier:
according to
Guideline:
OECD Guideline 215 (Fish, Juvenile Growth Test)
Version / remarks:
2000
Qualifier:
according to
Guideline:
EU Method C.14 (Fish Juvenile Growth Test)
Version / remarks:
2008
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: All test concentrations and stock solution
- Sampling method: The samples were taken in a 20 ml GC-vial that already contained 0.5 ml acetone.
- Sample storage conditions before analysis: none
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of stock solutions by loading two 10 L gas sampling bags each with 7 or 7.5 litres of ISO medium followed by addition of ~3.5 or 3.75 grams gas (500 mg/l). The gas sampling bags were closed immediately after addition and then shaken carefully for 4 hours on a platform shaker. Subsequently, the bags were left to settle overnight. The two saturated media (expected concentration ~160 mg/l) were mixed and then used for preparing the test concentrations by serial dilution.
Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Weight at study initiation (mean): 0.76 ± 0.08 g

FEEDING DURING TEST
- Food type: Cyprico Crumble Excellent (300-500 um), Coppens International bv, Helmond, The Netherlands).
- Amount: 4% of body weight per vessel
- Frequency: daily
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Hardness:
211.5 mg/L
Test temperature:
20.0 - 21.6 °C
pH:
6.9 - 7.9
Dissolved oxygen:
In freshly prepared solutions: 8.3 - 10.1 mg/L
In 24 h old solutions: 2.1 - 6.2 mg/L
In 24 h old control: 1.8 - 6.2 mg/L
Nominal and measured concentrations:
Nominal test substance concentrations: 1.5, 3.0, 6.0, 12 and 24 mg/L
Mean measured substance concentrations: 1.2, 2.7, 5.4, 11 and 22 mg/L (82 - 91% of nominal)
Details on test conditions:
TEST SYSTEM
- Test vessel: airthight glass vessels (40 L)
- Type: closed
- Fill volume: 38 L
- Aeration: none
- Renewal rate of test solution (frequency): daily
- No. of organisms per vessel: 16
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.32 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium, formulated using RO-water (tapwater purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands)

OTHER TEST CONDITIONS
- Photoperiod: 16:8 h day-night regime

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Daily determination of mortalities, clinical sings were determined three times a week, body weight was determined at test start and after 14 and 28 days of exposure respectively.

Reference substance (positive control):
yes
Remarks:
penta-chlorophenol
Duration:
28 d
Dose descriptor:
EC50
Effect conc.:
> 22 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
2.7 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
5.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Details on results:
- Type of and number with morphological abnormalities: none
- Type of and number with behavioural abnormalities: none
- Other biological observations: no mortalities observed
Statistical analysis on the growth rates for the interval of days 0-14 showed that the growth rate of the fish exposed to test groups of 5.4 and 22 mg/l were statistically significant lower than the growth of the fish in the control group.
Results with reference substance (positive control):
The reference test showed normal sensitivity of the supplied carp.
Reported statistics and error estimates:
ANOVA, Tukey test, Bonferroni t-test, TOXSTAT Release 3.5, 1996, D.D. Gulley, A.M. Boelter, H.L. Bergman

Table 1: Mean fish weights recorded at the start, after 14 and 28 days.

Day of recording

Mean fish weights (g):

Control

1.2 mg/L

2.7 mg/L

5.4 mg/L

11 mg/L

22 mg/L

0

0.76

0.79

0.78

0.77

0.78

0.73

14

1.11

1.11

1.10

1.06

1.10

1.01

28

1.49

1.48

1.49

1.40

1.45

1.27

% increase on day 28*

196

189

191

182

186

174

*Based on non-rounded figures.

Table 2: Mean growth rates recorded for the intervals of 0-14 days and 0-28 days.

Interval

Mean growth rates:

Control

1.2 mg/L

2.7 mg/L

5.4 mg/L

11 mg/L

22 mg/L

Day 0-14

2.71

2.45

2.48

2.22

2.47

2.32

Day 0-28

2.42

2.28

2.32

2.11

2.20

1.96

 

Description of key information

NOEC (28 d): 2.7 mg/L (measured; OECD 215)

Key value for chemical safety assessment

Additional information

The long-term toxicity of 2,3,3,3-Tetrafluoropropene to fish was tested according to OECD guideline 215 and in compliance with OECD Guidance Document No. 23 "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures" (Daikin Industries Ltd., 2011). Juvenile fish (Cyprinus carpio) were exposed to the substance in a semi-static system for 28 days. The test solutions were renewed on a daily basis and the test concentrations were verified analytically. Due to the gaseous test substance the test was performed in airtight vessels. The long-term toxicity of the substance was assessed by means of mortality, clinical symptoms and body weight of the test organisms. Mortality was not observed up to the highest tested concentration of 22 mg/L (measured). Based on the growth of the juvenile fish a NOEC of 2.7 mg/L (measured) was determined after 28 days of exposure.