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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
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, 1060, 2120, 3190, 4250 and 5310 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 (5310 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: 16.8 mm (SD=2.7)
- Weight at study initiation: 57 mg (SD=30)
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
44.9 mg CaCO3/L (SD=0.2)
Test temperature:
25.2ºC (SD=0.6)
pH:
7.5 (SD=0.3)
Dissolved oxygen:
6.1 mg O2/L (SD=0.9)
Salinity:
42.7 mg CaCO3/L (SD=0.7)
Nominal and measured concentrations:
Nominal concentrations: 0, 1060, 2120, 3190, 4250 and 5310 μg/L
Average measured concentrations: <195, 1050, 2090, 2720, 3640 and 4820 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 1050-4820 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: 2.85 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.:
3 150 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CL: 2920-3400 μg/L
Key result
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
1 480 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities: Affected fish swam with a corkscrew motion, were hypoactive and underreactive to external stimuli and were darkly colored. They also had convulsions and lost equilibrium prior to death.
- Mortality (96 hour) at <195, 1050, 2090, 2720, 3640 and 4820 μg/L (measured) were 0, 0, 0, 10, 90 and 100%, respectively.
- Effect (96 hour) at <195, 1050, 2090, 2720, 3640 and 4820 μg/L (measured) were observed in 0/10, 0/10, 10/10, 10/10, 10/10 and 10/10 fish respectively.
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
Sublethal observations / clinical signs:

Table 1: Mortalities

 

Hour

Control (0 µg/L)

A (1060 µg/L)

B (2120 µg/L)

C (3190 µg/L)

D (4250 µg/L)

E (5310 µg/L)

Initial

9

10

10

10

10

10

24

0

0

0

1

7

10

48

0

0

0

1

7

10

72

0

0

0

1

9

10

96

0

0

0

1

9

10

Table 2: Effects

Hour

Control (0 µg/L)

A (1060 µg/L)

B (2120 µg/L)

C (3190 µg/L)

D (4250 µg/L)

E (5310 µg/L)

Initial

9

10

10

10

10

10

24

0

0

0

10

10

10

48

0

0

10

10

10

10

72

0

0

10

10

10

10

96

0

0

10

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 alpha terpinene to fathead minnow (Pimephales promelas) was calculated to be 3150 (2920 -3400) 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 alpha terpinene at concentrations of <195, 1050, 2090, 2720, 3640 and 4820 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 <195, 1050, 2090, 2720, 3640 and 4820 μg/L (measured) were 0, 0, 0, 10, 90 and 100%, respectively. Treatment-related effects (96 hour) at <195, 1050, 2090, 2720, 3640 and 4820 μg/L (measured) were observed in 0/10, 0/10, 10/10, 10/10, 10/10 and 10/10 fish respectively. The 96 hour LC50, EC50 and NOEC of alpha terpinene to fathead minnow (Pimephales promelas) were calculated to be 3150 (2920 -3400) μg/L, 1480 μg/L and 1050 μg/L, respectively.

 

 

 

Description of key information

Key study: Test method similar to OECD 203. The 96h-LC50 of alpha terpinene to fathead minnow (Pimephales promelas) was found to be 3150 μg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
3.15 mg/L

Additional information