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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2005-2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes
Analytical monitoring:
not specified
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solution: 100 mg/L in 2-Methoxy ethanol. The test substance was diluted to 0.150 mg/L in the test water.
-.
Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Common name: ice fish
- Source: Nakajiama Fish Hatchery, Nagazu
- Length at study initiation: average 3.0 cm
- Weight at study initiation: avrage 0.24 g
.

ACCLIMATION yes
.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
not given
Test temperature:
24.5 - 25.1 deg C
pH:
7.7-7.8
Dissolved oxygen:
6.2 to 8.1
Salinity:
n.a.
Nominal and measured concentrations:
nominal 0.150 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: Round glass water tank, 4L
- Aeration:
- Renewal rate of test solution: every 8 to 16 h
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Well water, quality was confirmed according to several standards

.EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality
Reference substance (positive control):
yes
Remarks:
penta chlorphenol
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
> 0.15 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Results with reference substance (positive control):
LC50, 48 h: 0.642 mg/L
Validity criteria fulfilled:
not specified
Conclusions:
The test was performed at a concentration above the water solubility of the test substance. No mortality was observed. It can be concluded that the substance was not acutely toxic to fish at concentrations exceeding the water solubility.
Executive summary:

An acute 96 h LC50 test was performed with Oryzias Lapis as a toxicity screen for a bioconcentration study in carp. No mortality was observed up to the limit concentration tested in this study (0.15 mg/L). The low toxicity in fish is also confirmed in the low mortalitiy rate in the bioconcentration studies in rainbow trout and carp at the solubility limit that did not reveal any signs of toxicity at exposure durations up to 30 and 60 days respectively.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
August 1987 to February 1988
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The analytical monitoring has not been performed.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: U.S. Environmental Protection Agency (1975). Methods for acute toxicity testing with fish, macro-invertebrates and amphibians. EPA-660/3-75-009.
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES of the substance are all available in section 4 "Physical and chemical properties".
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Quantities of lonox 220 were added to five of the 6 glass aquaria to give a logarithmic series of concentrations between 10 and 1000 mg/L.
- Controls: One glass aquario without the testing substance.
- Chemical name of vehicle: No vehicle used.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The test substance appear to be out of solution at concentration greater than 10 mg/L in all of the test.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: S. gairneri
- Strain: Richardson
- Source: Fingerlings obtained from Itchin Valley Trout Farm, Hampshire (Batch No. RT 77).
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): a mean lenght of 3.6 cm (range 3.0 to 4.3 cm)
- Weight at study initiation (mean and range, SD): a weight of 0.81 g (range 0.39 to 1.07 g)
- Feeding during test: No

ACCLIMATION
- Acclimation period: 10 days
- Acclimation conditions (same as test or not): The fish are placed in 300 L circular self-cleaning fibreglass tanks at a density appropriate to their size. The tanks receive a continuous flow of temperature controlled water of a suitable and defined quality.
- Type and amount of food: Mainstream Trout Food No. 03 (BP Nutrition Ltd.).
- Feeding frequency: The quantities fed depending upon the size and age of the fish and the temperature of the water. Not fed in the 24h preceding the test.
- Health during acclimation (any mortality observed): Less tahn 5% over a 12 day period preceding the test

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
242-260 mg/L as CaCO3
Test temperature:
13-17°C
pH:
7.7-8.3
Dissolved oxygen:
9.6-10.4 mg/L
Nominal and measured concentrations:
Nominal concentration: logarithmic series of concentrations between 10 and 1000 mg/L.
0, 10, 30, 100, 300, 1000 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Six glass aquaria
- Material, size, headspace, fill volume: 10 L of filtered dechlorinated mains water
- Aeration: gently areated
- Renewal rate of test solution (frequency/flow rate): Daily renewal of the test solutions.
- No. of organisms per vessel: 10 fish placed in each aquaria

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory mains supply. The water comes from 2 pumping stations (Newnham and Wychling) controlled by the Mid Kent Water Company. The water is obtained from bore holes in the chalk of the North Downs. The only chemical treatment prior to its arrival in the
laboratory is chlorination to 0. 1 mg/L. The water was filtered (Balston LP-200-50-30) to remove parcicles larger than 10 µm and passed through activated carbon filter. (Cano model CT) to remove chlorine and organic contaminants .

After filtration and passage through activated carbon, the quality of the water is the following:
Note value is an overall range (13/10/80 - 19/3/87) (n = 26)
- Conductivity: 460 - 585
- pH: 7.1 -7.6
- SAT. INDEX: -0.3 - 0.1
- Total solid: 295 - 370
- Alkalinity: 247 - 275
- CARBON DIOXIDE: 9 - 23
- Total HARDNESS: 259 - 300
- Permanganate value: 0 .02 - <0.5
- Total chlorine: <0 .02 - <0 .05
- PHENOL: <5 - <5
- Total CALCIUM: 102 - 116
- Total MAGNESIUM: 2 .1 - 3
- Total IRON: <0.02 - 0 . 01
- Total MANGANESE: <0 .02 - <0.03
- Total MERCURY: <0.1 - <0.2
- Total LEAD: <5 - <50
- Total CADMIUM: <0 .5 - <5
- Total ARSENIC: <1 - <5
- Total COPPER: <0.03 - 0.11
- Total ZINC: <0.02 - <0.3
- Total SODIUM: 9 - 13
- Total POTASSIUM: 0.6 - 1.4
- CHLORIDE: 15 - 23
- Fluoride: <0.01 - <0. 1
- NITRATE: 2.9 - 11
- NITRITE: <1 - <1
- ORTHO PHOSPHATE: <0 .02 - 0.11
- Total SILICA: 10 - 12
- SULPHATE: 1 - 5
- AMMONIACAL N: <0.002 - 0.115
- ALBUMINOID N : <0.002 - 0.038

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: No data
- Light intensity: No data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
At 24h intervals the fish were observed and the number of fish exhibiting toxic symptoms were recorded. The symptoms were classified
into 5 categories:
a) No toxic symptoms
b) Fish swimming normally but exhibiting toxic symptoms (e.g. increased cough frequency, hyperventillation)
c) Fish swimming abnormally (e.g. on side or back)
d) Fish immobilised (e.g. lying on the bottom of thank or floating at surface, but still alive)
e) Fish dead

TEST CONCENTRATIONS
- Spacing factor for test concentrations: logarithmic series of concentrations between 10 and 1000 mg/L.
- Range finding study: No data
- Results used to determine the conditions for the definitive study: No data
Reference substance (positive control):
no
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
>= 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
>= 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
870 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% fiducial limits 630-1500 mg/L
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
820 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% fiducial limits 490-2600 mg/L
Details on results:
- Behavioural abnormalities:

* At 300 mg/L of the testing substance:
- After 24h of exposure --> 2 fish exhibiting toxic symptoms (b)
- After 48h of exposure --> 2 fish exhibiting toxic symptoms (b)
- After 72h of exposure --> 2 fish exhibiting toxic symptoms (b)
- After 96 h of exposure, one fish died (e) and 2 fish with toxic symptoms (b)
* At 1000 mg/L of the testing substance:
- After 24h of exposure --> 2 fish exhibiting toxic symptoms (b), 1 fish immobilized (d) and 3 fish died (e)
- After 48h of exposure --> 1 fish exhibiting toxic symptoms (b), 2 fish immobilized (d) and 5 fish died (e)
- After 72 h of exposure --> 1 fish exhibiting toxic symptoms (b), 1 fish immobilized (d) and 6 fish died (e)
- After 96 h of exposure --> 2 fish immobilized (d) and 6 fish died (e)

- Mortality of control: No
Results with reference substance (positive control):
No reference substance
Reported statistics and error estimates:
The 72 and 96h LC50 values (those concencrations causing a 50% mortality after 72 and 96h exposure) were calculated using probit analysis
after log. transformation of the concentrations (Finney, 1971).
Validity criteria fulfilled:
yes
Executive summary:

The acute toxicity of 4,4'-methylene-bis-2,6-di-tert-butyl phenol to the, rainbov trout, Salmo gairdneri was determined in a semi-static water test with daily renewal of the test solutions. Of the 10 fish exposed to 1000 mg/L of 4,4'-methylene-bis-2,6-di-tert-butyl phenol (the highest concentration tested),only 3 had died after 24h, and 5 after 48h. The 24 and 48h LC50 values were therefore >= 1000 mg/L. The 72 and 96h LC50 values were calculated to be 870 and 820 mg/L, respectively (95% fiducial limits 630-1500 mg/L and 490-2600 mg/L).

Description of key information

The acute toxicity for fresh water fish is above the water solubility of the test substance. 

Key value for chemical safety assessment

Additional information

Two acute fish studies with high and lower loading rates have demonstrates that the substance does not show any acute toxic effect at its solubility limit. Due to the low water solubility no LC50 value can be derived.

The low toxicity in fish is also confirmed in the low mortality rate in the bioconcentration studies in rainbow trout and carp at the solubility limit that did not reveal any signs of toxicity at exposure durations up to 30 and 60 days respectively.