Registration Dossier

Diss Factsheets

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17 April 1989 to 21 April 1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
equivalent or similar to guideline
Guideline:
EPA OPP 72-1 (Fish Acute Toxicity Test)
Version / remarks:
1982
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The product was added to the test water separately for each concentration by mixing with a small amount of water using an Ultra-Turrax stirrer. This mixture was then stirred into the aquarium.
Analytical monitoring:
yes
Details on sampling:
Analytical concentration controls were performed one hour after the beginning of the study and at the end of the study after about 96 hours.
Vehicle:
no
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: Bluegill
- Length at study initiation: 5.7 cm (range: 5.1 - 6.2 cm).
- Weight at study initiation: 2.1 g (range: 1.7 - 2.7 g).
- Corpulence factor of the batch: 1.2 (determined 05 April 1989). The corpulence factor (K) is calculated from the weight W (g) and the length L (cm) measured from the tip of the mouth to the distal end of the caudal fin according to the formula K = 100*W / L**3

ACCLIMATION
- Acclimation period: Adaptation to test water and test temperature was 14 days. The duration of housing and adaptation was about 2 months.
- Acclimation conditions: The fish were kept in a flow-through tank in tap water cleaned by active carbon and aerated with oil-free air. Temperature: 15 - 23 °C. Photoperiod: 16 hours of light and 8 hours of darkness. Total hardness about 2.5 mmol/L, acid capacity about 5.5 mmol/L, oxygen content > 60 % of maximum saturation and pH about 8.0.
- Type and amount of food during acclimation: ad libitum. Growing feed F/B 50, SSNIF Spezialdiaeten GmbH, Tetra Min standard feed for aquarium fish, tetra werke and life, or frozen artemia.
- Health during acclimation: Mortality during the last two weeks of housing: About 2.5 %. Mortality during the adaptation period: About 0.8 %. Fish were treated prophylactically as a routine measure 4 times with 0.05 mg/L malachite green chloride and 2 times with 10 mg/L tetracycline hydrochloride. The last treatment was more than 48 hours (19 days) before the start of the study. All fish used in the study were healthy.

FEEDING DURING TEST
- Food type: Food was withdrawn one day before and during exposure.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
2.2 mmol/L
Test temperature:
21 - 22 °C
pH:
About pH 8.0
Dissolved oxygen:
7.8 - 8.6 mg/L
Conductivity:
Max. 20 µOhm
Nominal and measured concentrations:
- Nominal concentrations: 0, 50 and 100 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass aquarium with a stainless-steel frame (80 x 35 x 46 cm)
- Material, size, headspace, fill volume: 100 L
- Aeration: Slight continuous aeration with oil-free air.
- Renewal rate of test solution: Static
- No. of organisms per vessel: 10 animals per replicate
- No. of vessels per concentration: Three for the high dose, one for the low dose
- No. of vessels per control: One
- Loading rate (g fish/ L test water): 0.2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted freshwater prepared from fully demineralised tap water. The re-salting was carried out by addition of 294 mg/L CaCL2·2H2O, 123.3 mg/L MgSO4·7H2O, 63 mg/L NaHCO3 and 5.5 mg/L KCl.
- Ca/mg ratio: 4:1
- Na/K ratio: 10:1
- Acid capacity: 0.8 mmol/L

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h darkness.

EFFECT PARAMETERS MEASURED:
Determination of calculation of the median lethal concentration (LC50) and, if possible, the LC5 and the LC95 using the Finney probit analysis after hours (nominal conc.) 1, 4, 24, 48, 72 and 96 h)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Based on the results of a range-finding study (neither mortality nor toxic signs at the conc. 100 mg/L) the concentration of 100 mg/L was selected and tested in three replicates according to the requirements of the EPA guideline (highest conc. to be tested). To be sure to obtain the NOEC the conc. 50 mg/L was tested in addition.
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 (fish)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
50 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: mortality and symptoms
Details on results:
- Abnormalities: Symptoms included apathy, convulsions, narcotic-like state and tumbling.
Sublethal observations / clinical signs:

Table 1: Mortality

Nominal Conc.

(mg/L)

No. of Fish

Dead Fish After

(h)

1

4

24

48

72

96

50

10

0

0

0

0

0

0

100

10

0

0

0

0

0

2

100

10

0

0

0

0

1

1

100

10

0

0

0

0

1

1

0

10

0

0

0

0

0

0

Table 2: Symptoms

Nominal Conc.

(mg/L)

No. of Fish

Symptoms After

(h)

1

4

24

48

72

96

50

10

100

10

T

T

NT

N

100

10

NT

AT

AT

100

10

KT

AT

T

0

10

T = Tumbling

N = Narcotic-like state

A = Apathy

K = Convulsions

Table 3: Analytically Derived Concentrations

Nominal Conc.

(mg/L)

Time (h)

1

96

50

46.1

48.3

100

94.2

96.5

100

96.3

96.3

100

98.5

97.7

0

0

0

 

Validity criteria fulfilled:
not specified
Conclusions:
Under the conditions of the study the 96 h LC50 was > 100 mg/L. The NOEC was 50 mg/L.
Executive summary:

The acute toxicity of the test material to the Bluegill sunfish (Lepomis macrochirus RAF) was assessed using a method which closely followed US EPA Guideline 72-1 and in compliance with GLP using a static system. 

Fish were exposed to nominal concentrations of 0, 50 and 100 mg/L for 96 hours. The high dose was tested in triplicate with 10 fish per replicate.

After 96 hours, at 100 mg/L 2 fish had died in one replicate and 1 had died per each of the remaining two replicates. Symptoms observed included apathy, convulsions, narcotic-like state and tumbling. The maximum concentration tested causing no mortality was 50 mg/L. The minimum concentration tested causing 100 % mortality was > 100 mg/L.

Under the conditions of the study the 96 h LC50 was > 100 mg/L. The NOEC was 50 mg/L. 

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 July 1984 to 13 July 1984
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
4th April 1984
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
The test concentrations were determined by UV-Spectroscopic method.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test material was added to the test water without any pre-treatment using an ultra-turrax stirrer. Adter 1 h the aquarium content was stirred again; subsequently the fish were placed in the aquaria.
Test organisms (species):
other: Salmo gairdneri Rich.
Details on test organisms:
TEST ORGANISM
- Common name: Rainbow trout
- Length at study initiation (length definition, mean, range and SD): 4.7 cm (range 4.5 - 4.9 cm)
- Weight at study initiation (mean and range, SD): 1.0g (range 0.8 - 1.2 g)
- Corpulance factor of animals consigned: 0.96. The corpulance factor "K" is calculated from the weight W (g) and the lenght "L" (cm) measured from the tip of the mouth to the end of the tail fin according to the formula:
K = 100 x G / Lx3
- Medical treatment: Once with 0.05 mg/L Malachite green choride.

ACCLIMATION
- Acclimation period: 7 weeks
- Acclimation conditions: The fish were kept in a flow-through tank in tap water cleaned and dechlorinated by active carbon and aerated with oil-free air.
- Type and amount of food during acclimation: "Fukosalm" growing feed and "Tetra Min" standard feed for aquarium fish.
- Feeding frequency during acclimation: Ad libitum.
- Health during acclimation (any mortality observed): 0 % mortality over 1 week before the beginning of the test.
- Water temperature: 15 - 18 ºC
- pH: About 7.5
- Oxygen content: > 60 % of air saturation value.
- Photoperiod: 16 h light and 8 h dark.


Test type:
static
Water media type:
freshwater
Remarks:
Reconstitued freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
2.5 mmol/L
Test temperature:
16 ± 1 ºC
pH:
7.8
Dissolved oxygen:
9.1 - 10.1 mg/L
Conductivity:
Maximum 10 µmho
Nominal and measured concentrations:
Nominal concentrations: 31.6, 46.4, 68.1, 100.0, 147.0, 215.0 and 316.0 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: Aquarium
- Material, size, headspace, fill volume: Glass aquarium with stainless steel frame (60 cm x 35 cm x 40 cm) and 50 L of volume.
- Aeration: Light continuous aeration with oil-free air.
- No. of organisms per vessel: 10
- Biomass loading rate: 0.2 g fish/ L test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted freshwater according to DIN 38 412, part 11, draft Sept. 1981. Preparation from fully demineralised tap water. The resalting is carried out by addition of:
294.0 mg/L CaCl2.2H2O
123.3 mg/L MgSO4.7H2O
64.8 mg/L NaHCO3
5.8 mg/L KCl
- Carbonate Hardness: 0.8 mmol/L
- Ratio Ca/Mg ions: 4:1
- Ratio Na/K ions: 10:1

EFFECT PARAMETERS MEASURED
- Mortality and symptoms. Determination of LC50 and, if possible, LC5 and LC95 using probit analysis after (nominal concentration) 1, 4, 24, 48, 72 and 96 h.

TEST CONCENTRATIONS
- Nominal concentrations: 31.6, 46.4, 68.1, 100.0, 147.0, 215.0 and 316.0 mg/L. An untreated control was also conducted.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
>= 150 - <= 220 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
68.1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
1 h
Dose descriptor:
LC50
Effect conc.:
>= 220 - <= 320 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
4 h
Dose descriptor:
LC50
Effect conc.:
>= 150 - <= 220 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
>= 150 - <= 220 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
>= 150 - <= 220 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
>= 150 - <= 220 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Other biological observations: The following symptoms were observed: Convulsions, discoloration, escape reflex and gasping. The symptom discoloration is regarded as a sign of stress and not as a sign of intoxication as it was also seen in the control group.
- Mortality:
1 h LC50: > 220 mg/L < 320 mg/L nominal concentrations (1 % significance level)
4 h LC50: > 150 mg/L < 220 mg/L nominal concentrations (1 % significance level)
24 h LC50: > 150 mg/L < 220 mg/L nominal concentrations (1 % significance level)
48 h LC50: > 150 mg/L < 220 mg/L nominal concentrations (1 % significance level)
72 h LC50: > 150 mg/L < 220 mg/L nominal concentrations (1 % significance level)
96 h LC50: > 150 mg/L < 220 mg/L nominal concentrations (1 % significance level)

No effect level observed: 68.1 mg/L
Maximum concentration causing no mortality: 147 mg/L
Minimum concentration causing 100 % mortality: 215 mg/L

- Any observations that might cause a difference between measured and nominal values: Undissolved test material was visible in all concentrations 1 h after beginning of the test. The amount increased with an increase in the concentration. After 24 h the amount of undissolved test compound has decreased and after 72 h only in the two highest concentrations small amounts were seen.
Results with reference substance (positive control):
None
Reported statistics and error estimates:
Statistics were conducted but details not specified.
Sublethal observations / clinical signs:

Mortality Results

Nominal Concentration

(mg/L)

Number of Fish Beginning

Number of Fish Dead

1 h

4 h

24 h

48 h

72 h

96 h

31.6

10

0

0

0

0

0

0

46.4

10

0

0

0

0

0

0

68.1

10

0

0

0

0

0

0

100.0

10

0

0

0

0

0

0

147.0

10

0

0

0

0

0

0

215.0

10

0

10

10

10

10

10

316.0

10

10

10

10

10

10

10

0

10

0

0

0

0

0

0

 

Symptoms

Nominal

Concentration

(mg/L)

Symptoms

1 h

4 h

24 h

48 h

72 h

96 h

31.6

 

 

 

 

V

 

46.4

 

 

 

 

 

 

68.1

 

 

 

 

 

 

100.0

 

 

VF

V

V

 

147.0

 

 

FVK

V

LV

 

215.0

L

 

 

 

 

 

316.0

 

 

 

 

 

 

0

 

 

 

 

V

 

F: Escape reflex

K: convulsions

L: Gasping

V: Discoloration

 

Analytically Detected Concentrations

Nominal

Concentration

(mg/L)

Analytically Detected Concentrations

(mg/L)

2 h

24 h

48 h

72 h

96 h

31.6

29

31

30

30

30

46.4

41

46

45

45

45

68.1

57

67

67

67

67

100.0

79

99

97

100

99

147.0

97

144

148

148

147

215.0

154

218

225

225

224

316.0

222

302

 

 

 

0

 

 

 

 

 

Validity criteria fulfilled:
not specified
Conclusions:
Under the conditions of the study the 96 h LC50 was: >150 mg mg/L <220 mg/L (1 % significance level) nominal. The no effect level observed was 68.1 mg/L.
Maximum concentration causing no mortality: 147 mg/L
Minimum concentration causing 100 % mortality: 215 mg/L
Executive summary:

The acute toxicity of the test material to fish was assessed according to OECD Test Guideline 203 and in compliance with GLP using a static method.

After 7 days acclimation the test was conducted on 10 rainbow trout per concentration at nominal concentrations of 31.6, 46.4, 68.1, 100.0, 147.0, 215.0 and 316.0 mg/L in glass aquaria with stainless steel frames.

Convulsions, discoloration, escape reflex and gasping were observed. The symptom discoloration is regarded as a sign of stress and not as a sign of intoxication as it was also seen in the control group.

Undissolved test material was visible in all concentrations 1 h after beginning of the test. The amount increased with an increase in the concentration. After 24 h the amount of undissolved test compound has decreased and after 72 h only in the two highest concentrations small amounts were seen.

Under the conditions of the study the 96 h LC50 was: >150 mg mg/L <220 mg/L nominal (1 % significance level). The no effect level observed was 68.1 mg/L.

Maximum concentration causing no mortality: 147 mg/L

Minimum concentration causing 100 % mortality: 215 mg/L

Description of key information

Munk (1989)

Under the conditions of the study the 96 h LC50 was > 100 mg/L. The NOEC was 50 mg/L.

Munk (1984)

Under the conditions of the study the 96 h LC50 was: >150 mg mg/L <220 mg/L nominal (1 % significance level). The no effect level observed was 68.1 mg/L.

Maximum concentration causing no mortality: 147 mg/L

Minimum concentration causing 100 % mortality: 215 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
100 mg/L

Additional information

Munk (1989)

The acute toxicity of the test material to the Bluegill sunfish (Lepomis macrochirus RAF) was assessed using a method which closely followed US EPA Guideline 72-1 and in compliance with GLP using a static system. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

Fish were exposed to nominal concentrations of 0, 50 and 100 mg/L for 96 hours. The high dose was tested in triplicate with 10 fish per replicate.

After 96 hours, at 100 mg/L 2 fish had died in one replicate and 1 had died per each of the remaining two replicates. Symptoms observed included apathy, convulsions, narcotic-like state and tumbling. The maximum concentration tested causing no mortality was 50 mg/L. The minimum concentration tested causing 100 % mortality was > 100 mg/L.

Under the conditions of the study the 96 h LC50 was > 100 mg/L. The NOEC was 50 mg/L. 

Munk (1984)

The acute toxicity of the test material to fish was assessed according to OECD Test Guideline 203 and in compliance with GLP using a static method. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

After 7 days acclimation the test was conducted on 10 rainbow trout per concentration at nominal concentrations of 31.6, 46.4, 68.1, 100.0, 147.0, 215.0 and 316.0 mg/L in glass aquaria with stainless steel frames.

Convulsions, discoloration, escape reflex and gasping were observed. The symptom discoloration is regarded as a sign of stress and not as a sign of intoxication as it was also seen in the control group.

Undissolved test material was visible in all concentrations 1 h after beginning of the test. The amount increased with an increase in the concentration. After 24 h the amount of undissolved test compound has decreased and after 72 h only in the two highest concentrations small amounts were seen.

Under the conditions of the study the 96 h LC50 was: >150 mg mg/L <220 mg/L nominal (1 % significance level). The no effect level observed was 68.1 mg/L.

Maximum concentration causing no mortality: 147 mg/L

Minimum concentration causing 100 % mortality: 215 mg/L