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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:
1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Study conducted similarly to OECD guideline 203 but without GLP compliance and with minor deviations (no data on acclimation period). The report is sufficiently well documented.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
No data on acclimation period
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0, 130, 260, 390, 520 and 650 μg/L
- Sampling time: 0, 24, 48, 72 and 96 hours
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test solutions was provided as liquid/liquid saturation units by adding excess of test chemical in 14 L of Lake Superior water
- Controls: Dilution water
- Chemical name of vehicle: water
- Concentration of vehicle in test medium: Stock (650 µg/L liq-liq equil.)
Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: Fathead minnows
- Source: ERL-Duluth, USA
- Age at study initiation: 30 days
- Length at study initiation: 20.9 mm (SD=2.2)
- Weight at study initiation: 128 mg (SD=35)
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
45.2 mg CaCO3/L (SD=0.3)
Test temperature:
25.2ºC (SD=0.2)
pH:
7.7 (SD=0.2)
Dissolved oxygen:
6.7 mg O2/L (SD=0.7)
Salinity:
42.2 mg CaCO3/L (SD=0.9)
Nominal and measured concentrations:
Nominal concentrations: 0, 130, 260, 390, 520 and 650 μg/L
Average measured concentrations: <31, 58, 140, 229, 303 and 424 μg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Six crystallizing dishes were used as test chambers in the electronic diluter with tank volumes of 200 mL; test solution depth: 4-5 cm
- Type (delete if not applicable): Closed
- Type of flow-through: Electronic diluter
- Renewal rate of test solution (frequency/flow rate): Flow rates of 21 mL/min to each tank produced toxicant concentrations of 58-424 μg/L
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): One
- No. of vessels per vehicle control (replicates): One
- Biomass loading rate: 6.4 g/L

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Lake Superior water
- Preparation of dilution water: Water was passed through sand filters, ultraviolet sterilizers and 55 µm fibre filter. Test waters were then aerated and heated at 25 ± 1 °C
- Intervals of water quality measurement: Tank temperatures (ºC), dissolved oxygen (mg/L) and pH were measured daily in each of the test chambers where test organisms were still alive

OTHER TEST CONDITIONS
- Photoperiod: 16 hour/day light cycle; cool white fluorescent light
- Light intensity: 12-28 lumens

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Behavior and morphological aberrations were observed at 0, 24, 48, 72 and 96 hours
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
280 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% CL: 259-303 μg/L
Key result
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
179 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities: Affected fish swam near the tank bottom, were hypoactive and underreactive to external stimuli, exhibited hemorrhaging and lost equilibrium prior to death.
- Mortality (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were 0, 0, 0, 0, 80 and 100%, respectively.
- Effect (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L μg/L (measured) were observed in 0/10, 0/10, 0/10, 10/10, 10/10 and 10/10 fish respectively.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Measured concentrations were lower than nominal concentrations due to volatility.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
- LC50 or EC50 values were calculated using the corrected average of the analyzed tank concentrations and the Trimmed Spearman-Karber Method (Hamilton et al., 1977).
- Mean toxicant concentrations used in the calculations were corrected for analytical spike recoveries

Table 1: Mortalities

 

Hour

Control (0 µg/L)

A (130 µg/L)

B (260 µg/L)

C (390 µg/L)

D (520 µg/L)

E (650 µg/L)

Initial

10

10

9

10

10

10

24

0

0

0

0

7

8

48

0

0

0

0

8

10

72

0

0

0

0

8

10

96

0

0

0

0

8

10

Table 2: Effects

Hour

Control (0 µg/L)

A (130 µg/L)

B (260 µg/L)

C (390 µg/L)

D (520 µg/L)

E (650 µg/L)

Initial

10

10

9

10

10

10

24

0

0

0

10

10

10

48

0

0

0

10

10

10

72

0

0

0

10

10

10

96

0

0

0

10

10

10

 

Validity criteria fulfilled:
yes
Remarks:
(mortality in control < 10%; flow-through test conditions; dissolved O2 concentration > 60% of air saturation value; results based on measured concentrations)
Conclusions:
The 96h-LC50 of d-alpha pinene to fathead minnow (Pimephales promelas) was calculated to be 280 (259 -303) μg/L.

Executive summary:

In an acute aquatic toxicity study performed similarly to OECD guideline 203, groups (10/dose) of fathead minnows (Pimephales promelas) were exposed to d-alpha pinene at concentrations of <31, 58, 140, 229, 303 and 424 μg/L (measured) for 96 hours under flow-through conditions. Mortality, behaviour and morphological aberrations were observed at 24, 48, 72 and 96 hours. All validity criteria were fulfilled.Mortalities (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were 0, 0, 0, 0, 80 and 100%, respectively. Treatment-related effects (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were observed in 0/10, 0/10, 0/10, 10/10, 10/10 and 10/10 fish respectively. The 96 hour LC50, EC50 and NOEC of d-alpha pinene to fathead minnow (Pimephales promelas) were calculated to be 280 (259 -303) μg/L, 179 μg/L and 140 μg/L, respectively.

 

 

 

Description of key information

Key study: Test method similar to OECD TG 203. The 96h-LC50 of d-alpha pinene to fathead minnow (Pimephales promelas) was calculated to be 280 (259 -303) μg/L.

Key value for chemical safety assessment

LC50 for freshwater fish:
0.28 mg/L

Additional information

Key study: In an acute aquatic toxicity study performed similarly to OECD guideline 203, groups (10/dose) of fathead minnows (Pimephales promelas) were exposed to d-alpha pinene at concentrations of <31, 58, 140, 229, 303 and 424 μg/L (measured) for 96 hours under flow-through conditions. Mortality, behaviour and morphological aberrations were observed at 24, 48, 72 and 96 hours. All validity criteria were fulfilled. Mortalities (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were 0, 0, 0, 0, 80 and 100%, respectively. Treatment-related effects (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were observed in 0/10, 0/10, 0/10, 10/10, 10/10 and 10/10 fish respectively. The 96 hour LC50, EC50 and NOEC of d-alpha pinene to fathead minnow (Pimephales promelas) were calculated to be 280 (259 -303) μg/L, 179 μg/L and 140 μg/L, respectively.