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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study is classified as Klimisch Code 2 (reliable w ith restriction) as it was performed to an internationally recognised method (OECD 203) and was conducted to Good Laboratory Practice. However, temperature deviations were reported during the acclimisation period, the homogeneity and stability of the test material in the test preparations were not determined and an error in preparation of fresh test media for dosing at 24 hours resulted in a change from a semi-static to static study.
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
temperature deviations were reported during the acclimisation period and there was an error in preparation of fresh test media for dosing at 24 hours
GLP compliance:
yes
Analytical monitoring:
no
Details on sampling:
- Concentrations: 0 and 1000 mg/l nominal WAF loading rate
- Sampling method: The concentration, homogeneity and stability of the test material in the test preparations were not determined. Total Organic Carbon (TOC) analysis was performed on the test preparations at 0, 24, 48, 72 and 96 hours. Duplicate samples were taken and stored at -20°C for further analysis if necessary.
- Sample storage conditions before analysis: Not reported.
Vehicle:
no
Details on test solutions:
PREPARATION OF SYNTHETIC SEAWATER
Tropic Marin(R) Synthetic Sea Salt (5100 g) was dispersed in a volume of 200 litres of water, giving a nominal salinity of 22 ppt. The pH was adjusted to 8.0 using 10 M HCl.

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 21.00 g of test material was added to the surface of 21 litres of synthetic seawater. The synthetic seawater was then stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the w ater surface. Stirring was conducted for 16 hours. The mixture was then allowed to stand for 4 hours. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel to a depth of 5 cm from the bottom of the vessel. A length of Tygon tubing w as inserted into the glass tube and pushed through the Nescofilm seal. The aqueous phase or WAF was removed by mid-depth siphoning (the first 75-100 ml discarded) to give the 1000 mg/l loading rate WAF.
- Evidence of undissolved material: Microscopic inspection of the WAF showed no micro-dispersions or undissolved test material to be present.
- Observations on test material solubility: At the start and end of the mixing period, and after the 4 hour settlement period, the test media was observed to be a clear colourless water column w ith a dark brown poly slick of test material on the surface. After siphoning, and for the test duration, the test media was observed to be a clear, colourless solution.
Test organisms (species):
Cyprinodon variegatus
Details on test organisms:
TEST ORGANISM
- Common name: Sheepshead minnow
- Source: Aquatic Research Organisms, New Hampshire, USA and maintained in-house since 29 July 2005 in a glass tank w ith an activated carbon recirculation system
- Age at study initiation: Juvenile
- Length at study initiation: Mean standard length of 1.8 cm (SD = 0.3)
- Weight at study initiation: Mean weight of 0.22 g (SD = 0.6)
- Feeding during test: Fish were not fed during the test

ACCLIMATION
- Acclimation period: Fish were acclimitised to test conditions from 24 November 2005 to 6 December 2005
- Acclimation conditions: Same as test conditions
- Type and amount of food: Commercial Tetramin(R) flake food, which was discontinued approximately 24 hours prior to the test
- Feeding frequency: Not stated
- Health during acclimation: Less than 1% mortality occurred in the 7 days prior to the test
Test type:
other: semi-static for the first 24 hours, static for the remainder of the test
Water media type:
saltwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
Not reported
Hardness:
Not relevant
Test temperature:
Refer to Table 1

During the acclimitisation period, temperate was controlled at 19.1°C to 23.6°C, although it was reported to be above the 18°C to 22°C range on three days. This deviation was not considered to affect the validity or intergrity of the study on the basis that zero mortality was observed during the acclimitisation period.
pH:
Refer to Table 1
Dissolved oxygen:
Refer to Table 1
Salinity:
Refer to Table 1
Nominal and measured concentrations:
0 and 1000 mg/l nominal WAF loading rate
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material: 20 litre glass vessel, covered to reduce evaporation
- Aeration: Aerated via narrow bore glass tubes
- Renewal rate of test solution: Due to a technical error, preparation of fresh test media for dosing at 24 hours w as incorrectly performed. Therefore, it w as not possible to use the freshly prepared test media at 24 hours. To avoid the unnecessary use of additional test organisms for ethical reasons, testing was continued under a static regime as opposed to using semi-static conditions
- No. of organisms per vessel: Ten (Three in the range-finding study)
- No. of vessels per concentration (replicates): Two (One in the range-finding study)
- No. of vessels per control (replicates): One
- Biomass loading rate: 0.11 g/l

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Synthetic seawater. Tropic Marin(R) Synthetic Sea Salt (5100 g) was dispersed in a volume of 200 litres of water, giving a nominal salinity of 22 ppt. The pH was adjusted to 8.0 using 10 M HCl
- Total organic carbon: Refer to Table 2
- Intervals of water quality measurement: 0, 24, 48, 72 and 96 hours

OTHER TEST CONDITIONS
- Adjustment of pH: Not stated
- Photoperiod: 16 hour light, 8 hour darkness w ith 20 minute dawn and dusk transitions
- Light intensity: Not stated

EFFECT PARAMETERS MEASURED: Mortality and any sublethal effects of exposure, determined by visual inspection after 3, 24, 48, 72 and 96 hours

TEST CONCENTRATIONS
- Test concentrations: 0 and 1000 mg/l nominal WAF loading rate
- Range finding study
- Test concentrations: 0 and 1000 mg/l nominal WAF loading rate
- Results used to determine the conditions for the definitive study: No mortality and no sub-lethal effects were noted in the range-finding test.
Reference substance (positive control):
no
Key result
Duration:
3 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
3 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
6 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
6 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
24 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
24 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
48 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
72 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Details on results:
No mortalities occured at a concentration of 1000 mg/l nominal WAF loading rate and no sub-lethal effects were noted after 96 hours.
Reported statistics and error estimates:
Statistical analysis of the data was not warranted on the basis that no mortality occurred in this study.
Validity criteria fulfilled:
yes
Conclusions:
The test substance was not toxic to the Sheepshead minnow (Cyprinidon variegatus) at a loading rate of 1000 mg/L nominal WAF.
Executive summary:

In a 96-hour acute toxicity study, Sheepshead minnow (Cyprinidon variegatus) were exposed to Cardolite NC-603at nominal concentrations of 0 and 1000 mg/L WAF under static conditions. The 96-hour LL50w as >1000 mg/L nominal WAF. No signs of toxicity w ere noted. This toxicity study is classified as acceptable and satisfies the guideline requirement for a fish acute toxicity study.

Results Synopsis:

Test organism size/age (mean w et w eight or length): Juvenile, mean standard length of 1.8 cm (SD = 0.3), mean w eight of 0.22 g (SD = 0.6)

Test Type: Static

LL50: >1000 mg/L nominal WAF

NOELR: 1000 mg/L nominal WAF

Endpoint(s) Effected: None

Description of key information

Key study: Test method OECD 203. GLP study. The 96h-LL50 was determined to be > 1000 mg/L nominal WAF in Sheepshead minnow.

Key value for chemical safety assessment

LC50 for marine water fish:
1 000 mg/L

Additional information

Key study: An acute toxicity on Sheepshead minnow (Cyprinidon variegatus) was assessed according to OECD 203. Ten fish were exposed to 1000 mg/L nominal WAF (based on preliminary range-finding results) over an exposure period of 96 hours under static conditions. The 96h-LL50 was determined to be > 1000 mg/L nominal WAF. No sign of toxicity were noted.