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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:
From October 19, 2015 to October 29, 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
no
Details on sampling:
Analytical samples were taken from the threshold concentration and control in 24 h intervals.
Vehicle:
no
Details on test solutions:
TEST SUBSTANCE SOLUTION PREPARATION
The necessary amount of test substance for preparing the stock solution was weighed on a weighing scoop and transferred to a volumetric flask. Test medium was added up to the bench mark and the solution was homogenised by shaking. The solution was turbid. The solution was then applied to the respective aquarium filled with test medium. Special care was taken to ensure the complete transfer of the test substance into the test vessel. The final solution in the aquarium was clear, no turbidity was observed at test start. After 48 h, the test solution appeared turbid and became more turbid each day.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Species: Oncorhynchus mykiss Walbaum (Salmoniformes: Salmonidae)
- Source: Forellenzucht Störk, D-88348 Bad Saulgau, Germany
- Length at study initiation: 4 - 6 cm
- Feeding: each day
- Food type: granular rearing food
- Amount: 2% of bodyweight

ACCLIMATION
- Acclimation period: >12 d
- Health during acclimation: No mortality above 5% was observed throughout the acclimatisation period


Test type:
static
Limit test:
yes
Total exposure duration:
96 h
Hardness:
8°dH corresponding to 143 mg CaCO3/L
Test temperature:
15 to 17°C
pH:
7.77 - 8.22
Dissolved oxygen:
The dissolved oxygen was ≥88% of air saturation.
Details on test conditions:
TEST SYSTEM
- Test vessel: 25 L glass aquaria
- Fill volume: 15 L
- Aeration: Continuous aeration of the test tanks with a membrane pump using a Pasteur pipette.
- No. of animals per concentration: 7 (no replicates)
- Loading: maximum loading of 1 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: water composed of dechlorinated drinking water and deionised water.
- Conductivity: 363 µS/cm
- pH of the aerated water: 6.0-8.5
- Dissolved oxygen: above 60% saturation
- Total hardness: 140-250 mg/L (as CaCO3), corresponding to 7.8-14°dH.
- Photoperiod: 12 to 16 h daily

EFFECT PARAMETERS MEASURED
Observations
Fish were observed at 0, 4, 24, 48, 72 and 96 h after test start. Fish were considered dead if there was no visible movement (e.g. gill movement), and if touching of the caudal peduncle produced no reaction. Records were made on visible abnormalities as: loss of equilibrium, swimming behaviour, respiratory function, pigmentation and all other observed events. At termination of the test, all fish were euthanized and then weighed and measured.

Measurements
Measurements of temperature, pH-value and oxygen saturation were performed in 24 h intervals. Water hardness of the untreated control was determined at the beginning of the test.

TEST CONCENTRATIONS
0 and 100 mg/L
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
In the control and at 100 mg/L, no mortality was observed within the period of the test. No sublethal effects were observed in the control and at the threshold concentration of 100 mg/L. The average weight of the test organisms was 0.94±0.18 g; the average length was 49±3 mm.
Reported statistics and error estimates:
Since no mortality occurred during the test, statistical evaluation for the estimation of the LC50 was not required.

The test substance is a complex mixture of various organic ingredient groups having substantially different chemical structures and the toxicity cannot be assigned to a special ingredient. Consequently, no appropriate analytical method for an accurate analysis was available. However, as approximation of the content of test substance in aqueous media, the content of total organic carbon (TOC) was measured. The content of TOC in the test substance was analysed to be 56%.

Table 2: TOC measurements during the test

Test substance nominal [mg/L]

TOC content nominal (56%)
[mg/L]

Sampling
[h],

TOC [mg/L]

TOC
[%]

Measured in test solution

Measured in test solution (control corrected)

Control

-

0 (fresh)

0.6

-

-

-

96 (aged)

1.6

-

-

100

56.0

0 (fresh)

55.0

54.4

97

56.0

96 (aged)

54.7

53.1

95

 

Validity criteria fulfilled:
yes
Conclusions:
The 96 h LC50 of the test substance in Rainbow trout (Oncorhynchus mykiss ) was >100 mg/L (nominal) and the 96 h NOEC was 100 mg/L.
Executive summary:

A study was conducted to determine the acute toxicity of the test substance in Rainbow trout (Oncorhynchus mykiss) under static conditions according to OECD Guideline 203, in compliance with GLP. The test substance was evaluated in a limit test at a threshold concentration of 100 mg/L. Test solution was prepared by dilution of the substance in medium and application of a defined volume of the stock solution to the test vessel. Assessments on effects and mortality were conducted after 0, 4, 24, 48, 72 and 96 h. Samples were taken at 0 h (initial value) and after 96 h from aged test solutions to determine total organic carbon content (TOC analysis). No sublethal effects were observed at 0 and 100 mg/L (nominal). The absence of mortality at the threshold concentration indicates that the fish is not the most sensitive group of test organism and that the LC50 is greater than the threshold concentration.TOC analysis showed that the test item was stable throughout the period of the test. The initial measured content of TOC was 98 % of the nominal concentration. The measured content of TOC in aged test solution was 95 % of the nominal concentration.Under the study conditions, the 96 h LC50 of the test substance in Rainbow trout was >100 mg/L (nominal) (Tanneberger C, 2015).

Description of key information

Key value for chemical safety assessment

LC50 for freshwater fish:
100 mg/L

Additional information

A study was conducted to determine the acute toxicity of the test substance in Rainbow trout (Oncorhynchus mykiss) under static conditions according to OECD Guideline 203, in compliance with GLP. The test substance was evaluated in a limit test at a threshold concentration of 100 mg/L. Test solution was prepared by dilution of the substance in medium and application of a defined volume of the stock solution to the test vessel. Assessments on effects and mortality were conducted after 0, 4, 24, 48, 72 and 96 h. Samples were taken at 0 h (initial value) and after 96 h from aged test solutions to determine total organic carbon content (TOC analysis). No sublethal effects were observed at 0 and 100 mg/L (nominal). The absence of mortality at the threshold concentration indicates that the fish is not the most sensitive group of test organism and that the LC50 is greater than the threshold concentration.TOC analysis showed that the test item was stable throughout the period of the test. The initial measured content of TOC was 98% of the nominal concentration. The measured content of TOC in aged test solution was 95% of the nominal concentration. Under the study conditions, the 96 h LC50 of the test substance in Rainbow trout was >100 mg/L (nominal) (Tanneberger C, 2015).