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Ecotoxicological information

Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from January 31, 2003 to April 30, 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study has been conducted according to the appropriate OECD guideline, under GLP and analytical monitoring was carried out.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Remarks:
Cetrificate is not included in the study report.
Analytical monitoring:
yes
Details on sampling:
Test solution;
The test solution of 0.75 mL was collected from each test vessel at the start of exposure and 24 hours after exposure (before water renewal). The equivalent amount of acetone was added and mixed, and the solution was analyzed by GC.
Vehicle:
no
Details on test solutions:
Test substance solution;
The test substance of 500 mg (the test substance of 475 µL) was sampled and added in dilution water, and then made to constant volume of 5000 mL with dilution water to give 100 mg/L test substance concentration. This test solution was prepared at every water exchange.
Control solution;
Control solution was used only dilution water without adding test substance.
Test organisms (species):
Oryzias latipes
Details on test organisms:
Test fish;
Japanese name: Himedaka
Scientific name: Oryzias latipes
Supplier: Tsunashima fishing (18-1, Tsunashima-Nishi 5-chome, Kohoku-ku, Yokohama-shi, Kanagawa)
Receipted date: July 29, 2002
Body length: mean; 2.29 cm (2.19 - 2.40 cm), n = 10
Body weight: mean; 0.197 g (0.161 - 0.229 g), n = 10
Sensitivity: LC50-96 hr = 1.2 mg/L (95%CI = 0.81 - 1.7 mg/L), Copper sulphate•5H2O
This value almost corresponds to LC50 in Yokohama Laboratory as follows (n = 23, since 1995);
Mean ± standard deviation = 0.92 ± 0.29 mg/L
Minimum to maximum value = 0.44 to 1.5 mg/L
Age: Approximately 1 year after hatching
Acclimatization period: From February 21, 2003 to March 10, 2003
The 7-day mortality before the starting of exposure was less than 5%, and the visually healthy and normal fish were used for the study. Medical bath was not performed during the acclimatization.
Acclimatization conditions for 14-day acclimatization before the start of exposure;
Rearing water: Dilution water
Rearing method: Flow-through method (rearing density; less than 1.0 g/L/day)
Water temperature: 24 ± 1°C
Concentration of dissolved oxygen: More than 80% of saturated concentration
pH: 6.5 - 8.5
Lighting: Under room light, 16 hours light (less than 1000 lux) / 8 hours dark
Feeding: Tetramin (Tetraberk)
Feed amount: Approximately 2% of fish body weight (no feeding in 24 hours before the start of exposure)
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
Dilution water;
Total hardness (as CaCO3): 73 mg/L
Test temperature:
(unit: °C)
Control; 0 hour (new: 23.8), 24 hours (old: 24.1, new: 23.8), 48 hours (old: 24.0, new: 23.9), 72 hours (old: 23.8, mew: 23.6), 96 hours (old: 23.8)
Test solution; 0 hour (new: 23.8), 24 hours (old: 24.1, new: 23.8), 48 hours (old: 24.0, new: 23.9), 72 hours (old: 23.8, mew: 23.6), 96 hours (old: 23.8)
(Mean* measured concentration of test solution: 101 mg/L)
*; geometric mean
pH:
Control; 0 hour (new: 7.6), 24 hours (old: 7.5, new: 7.8), 48 hours (old: 7.4, new: 7.5), 72 hours (old: 7.5, mew: 7.7), 96 hours (old: 7.5)
Test solution; 0 hour (new: 7.6), 24 hours (old: 7.5, new: 7.6), 48 hours (old: 7.5, new: 7.6), 72 hours (old: 7.4, mew: 7.8), 96 hours (old: 7.4)
(Mean* measured concentration of test solution: 101 mg/L)
Dissolved oxygen:
(unit: mg/L)
Control; 0 hour (new: 8.4), 24 hours (old: 6.7, new: 8.3), 48 hours (old: 6.2, new: 8.4), 72 hours (old: 6.4, mew: 8.5), 96 hours (old: 6.2)
Test solution; 0 hour (new: 8.4), 24 hours (old: 6.6, new: 8.4), 48 hours (old: 6.4, new: 8.5), 72 hours (old: 6.5, mew: 8.5), 96 hours (old: 6.1)
(Mean* measured concentration of test solution: 101 mg/L)
Salinity:
No description
Nominal and measured concentrations:
Nominal test substance concentration; 100 mg/L
Measured test substance concentration; see Table 3
Details on test conditions:
(1) Exposure method; Semi-static method (The whole volume of test solution was renewed at every 24 hours)
(2) Exposure period; 96 hours
(3) Volume of test solution; 5.0 L/vessel
(4) Replicate; One vessel/test concentration area
(5) Number of test fish; 10 fish/test concentration area
(6) Test temperature; 24 ± 1°C
(7) Concentration of dissolved oxygen; Maintained more than 60% of saturated concentration (no aeration)
(8) pH; No pH adjustment of test solution was performed.
(9) Lighting; Under room light, 16 hours, light (less than 1000 lux), 8 hours, dark
(10) Feeding; None
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 101 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: The calculation of 95% confidence interval was impossible
Details on results:
See Table 1 and 2
Sublethal observations / clinical signs:

Table 0 : Result of range finding study

Concentration(mg/L)

96hr-Mortality(%)

100

0

*n=1, 5 fish/vessel

Table 1 : Mortality of the Medaka

Nominal

concentration

(mg/L)

Mean* measured concentration

(mg/L)

Cumulative mortality (Percent mortality)

24

hours

48

hours

72

hours

96

hours

Control

---

0 (0)

0 (0)

0 (0)

0 (0)

100

101

0 (0)

0 (0)

0 (0)

0 (0)

*; geometric mean

Table 2 : Observed toxicological symptoms

Nominal

concentration

(mg/L)

Mean* measured concentration

(mg/L)

Symptoms (Symptom-number of fish)

24

hours

48

hours

72

hours

96

hours

Control

---

N

N

N

N

100

101

N

N

N

N

*; geometric mean

N; No toxicological symptom was observed.

Calculation of Lethal Concentration 50 (LC50)

The mortality (%) was usually calculated from the number of death of medaka and the number of the tested individuals (10 fish) at each test area at 24, 48, 72 and 96 hours after the start of exposure to determine median lethal concentration (LC50). However, median lethal concentration (LC50) was not calculated because this test was a limit test only at the upper test concentration limit and “> the upper test concentration limit” was adopted.

Validity criteria fulfilled:
yes
Conclusions:
A 96 h LC50 value of >101 mg/l has been determined for the effects of the test substance on mortality of the freshwater fish Oryzias latipes. The result is based on geometric mean measured concentration.

Description of key information

96 h,  LC50 >101 mg/L, Oryzias latipes, measured.

Key value for chemical safety assessment

Additional information

A 96 h LC50 value of >101 mg/l has been determined for the effects of the test substance on mortality of the freshwater fish Oryzias latipes. The result is based on geometric mean measured concentration.