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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
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Guideline study but non-standard species; nominal toxicity values: not reliable
Qualifier:
according to guideline
Guideline:
other: ASTM E-729 (1980)
Version / remarks:
American Society for Testing and Materials (1980) Standard practice for conducting acute toxicity tests with fishes, macroinvertebrates, and amphibians. American Society for Testing Materials Report E-729, Philadelphia, PA, 25 pp
Deviations:
yes
Remarks:
use of various dilution waters
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test solutions were prepared by either pipetting appropriate aliquots of stock solution prepared in deionized water on the day of use or adding the compound directly to the test vessel.
Test organisms (species):
Oncorhynchus tschawytscha
Details on test organisms:
TEST ORGANISM
- Common name: Chinook salmon
- Source: Fall-run chinook salmon were obtained as eyed eggs from Merced River Fish Facility, Snelling, CA, in 1986.
- Method of breeding: Following hatching, swim-up fry and older fish were cultured according to procedures described by Brauhn and Schoettger (1975).

ACCLIMATION
- Type and amount of food: Fish were fed a commercial salmon diet (Biodiet®)
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
211 +/- 1 mg CaCO3/L
Test temperature:
12 +/- 1 °C
pH:
7.82 +/- 0.35
Nominal and measured concentrations:
Nominal concentrations were expressed as the total element added as determined from the certificate of analysis.
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 19.6 L glass jars containing 15 L of test solution
- Aeration: not specified
- No. of organisms per vessel: 10 or 2 x 5 for large, advanced fry
- Biomass loading rate: < 0.8 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Standardized San Luis Drain water (SLD) without trace elements, was diluted 10-fold in a standardized fresh receiving water (SLD/fresh water). For further information please see "Any other information on materials and methods incl .tables".
- Alkalinity: 87.2 +/- 0.8 mg CaCO3/L
- Ca/mg ratio: 45.8 +/- 0.8 / 23.6 +/- 0.5 mg/L
- Conductivity: 721 +/- 9 µmhos/cm @ 25 °C


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations on mortality and abnormal behavioral responses were made at 24 h intervals and all dead fish removed after each observation.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
725 mg/L
Nominal / measured:
nominal
Conc. based on:
element
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CI = 590 - 840 mg/L
Reported statistics and error estimates:
The method of Litchfield and Wilcoxon (1949) was used to calculate 96 h LC50 values, 95% confidence intervals, slope function values, and to test for parallelism of concentration mortality curves. The Standard Error of the Difference as described by Sprague and Fogels (1977) and Zar (1974) was used to determine statistical differences in LC50 values.
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Peer reviewed study
Qualifier:
no guideline followed
Principles of method if other than guideline:
The acute toxicity of boron has been studied with the dab (Limanda limanda) by subjecting them to a range of concentrations of the test material for 96 h in a continuous flow system. No standard guideline was identified, but methods were equivalent to standard fish toxicity testing methods.
GLP compliance:
yes
Remarks:
OECD, 1982
Analytical monitoring:
yes
Vehicle:
no
Test organisms (species):
Limanda limanda
Details on test organisms:
TEST ORGANISM
- Common name: Dab
- Source: Uncontaminated south Devon environments
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): mean length of 10.1 cm (range 7.0 - 13.2 cm)
- Weight at study initiation (mean and range, SD): mean weight of 16.9 g (range 5.6 - 30.8 g)
- Feeding during test: No feeding during test


ACCLIMATION
- Acclimation period: > 7 days in the laboratory, then acclimated to the test temperature of 12°C at a rate of 1-3 °C per day from their ambient temperature storage conditions. They were then maintained at the test temperature for a minimum of three days before being transferred to the test system where they were held for a minimum of 24 h under test conditions prior to the commencement of the exposure to the test substance.
- Feeding frequency: No feeding during acclimatisation
Test type:
flow-through
Water media type:
saltwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
12 +-1 °C
pH:
7.7 +/- 0.8
Dissolved oxygen:
7.9 +/- 0.6 mg/L
Salinity:
34.6 +/- 0.2 ‰
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 30 L rectangular glass tanks
- Aeration: The test solutions were gently aerated with compressed air.
- Renewal rate of test solution (frequency/flow rate): 24h
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: 24h
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
74 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element
Basis for effect:
mortality (fish)
Remarks on result:
other: 66.4-83.0 mg/L
Reported statistics and error estimates:
Probit analysis (Finney, 1971), of mortality observations every 24 h and measured test substance concentration gave the LC50 values for each substance.
Sublethal observations / clinical signs:

LC50 after 24h : 88.3 (80.3 - 97.3) mg/L

LC50 after 48h : 75.7 (68.0 - 85.0) mg/L

LC50 after 72h : 75.7 (68.0 - 85.0) mg/L

Conclusions:
The study was not conducted according to standard guidelines, but methods are equivalent to standard fish toxicity testing methods. Hence, the method is to be considered scientifically reasonable and the results can be considered as reliable to assess the toxicity towards fish. The test was conducted on the test material Disodium tetraborate.
The toxic effect towards fish was assessed on the saltwater species Dab (Limanda limanda) in a 96 h concentration-response test with flow-through conditions. The 96-hour LC50 was determined to be 74 mg/L based on the boron element. The exposure concentrations were verified by analytical measurements.
Executive summary:

The study was not conducted according to standard guidelines, but methods are equivalent to standard fish toxicity testing methods. The test was conducted on the test material Disodium tetraborate.

The purpose of this study was to determine the toxicity of the test item to fish. For this purpose, the saltwater species Dab (Limanda limanda) was exposed in a static test to various concentrations under defined conditions for 96 hours. The study encompassed 6 treatment groups (5 dose rates of the test item and a control) each containing 20 individuals.The exposure concentrations were verified by analytical measurements. The mortality of the fish was determined by visual observation after 24, 48, 72 and 96 hours.

The 96h-LC50 was determined to be 74 mg/L based on the boron element.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Guideline study, but non-standard species; nominal toxicity values: not reliable
Qualifier:
according to guideline
Guideline:
other: ASTM E-729 (1980)
Version / remarks:
American Society for Testing and Materials (1980) Standard practice for conducting acute toxicity tests with fishes, macroinvertebrates, and amphibians. American Society for Testing Materials Report E-729, Philadelphia, PA, 25 pp
Deviations:
yes
Remarks:
use of various dilution waters
GLP compliance:
not specified
Remarks:
not specified
Analytical monitoring:
not specified
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test solutions were prepared by either pipetting appropriate aliquots of stock solution prepared in deionized water on the day of use or adding the compound directly to the test vessel.
Test organisms (species):
Oncorhynchus tschawytscha
Details on test organisms:
TEST ORGANISM
- Common name: Chinook salmon
- Source: Fall-run chinook salmon were obtained as eyed eggs from Merced River Fish Facility, Snelling, CA, in 1986.
- Weight at study initiation (mean and range, SD): 0.31g
- Method of breeding: Following hatching, swim-up fry and older fish were cultured according to procedures described by Brauhn and Schoettger (1975).

ACCLIMATION
- Type and amount of food: Fish were fed a commercial salmon diet (Biodiet®).
Test type:
static
Water media type:
freshwater
Remarks:
standard soft water prepared as recommended by the U.S. Environmental Protection Agency (USEPA) for use in acute toxicity tests with fish (USEPA 1975)
Limit test:
no
Total exposure duration:
96 h
Hardness:
41.7 +/-0.3 mg CaCO3/L
Test temperature:
12 +/- 1 °C
pH:
7.57 +/- 0.06
Nominal and measured concentrations:
Nominal concentrations were expressed as the total element added as determined from the certificate of analysis.
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 19.6 L glass jars containing 15 L of test solution
- Aeration: Not specified.
- No. of organisms per vessel: 10 or 2 x 5 for large, advanced fry
- Biomass loading rate: < 0.8 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Standard soft water prepared as recommended by the U.S. Environmental Protection Agency (USEPA) for use in acute toxicity tests with fish (USEPA 1975). For further information please see "Any other information on materials and methods incl. tables".
- Alkalinity: 29.6 +/- 0.6 mg CaCO3/L
- Ca/mg ratio: 7/6 mg/L
- Conductivity: 157 +/- 3 µmhos/cm @ 25 °C

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations on mortality and abnormal behavioral responses were made at 24 h intervals and all dead fish removed after each observation.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
566 mg/L
Nominal / measured:
nominal
Conc. based on:
element
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CI = 482 - 664 mg/L
Reported statistics and error estimates:
The method of Litchfield and Wilcoxon (1949) was used to calculate 96 h LC50 values, 95% confidence intervals, slope function values, and to test for parallelism of concentration mortality curves. The Standard Error of the Difference as described by Sprague and Fogels (1977) and Zar (1974) was used to determine statistical differences in LC50 values.
Sublethal observations / clinical signs:

Results

Acute toxicity (LC50s in mg/L; 95% confidence intervals in parentheses)

of boron to different life stages of chinook salmon a in soft water

Life stage or weight [g]

Exposure Duration

24 h

96 h

eyed egg

> 1000

> 1000

alevin

> 1000

> 1000

0.3

> 1000

566

(95% CI = 482 – 644)

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Guideline study but non-standard species; nominal toxicity values: not reliable
Guideline:
other: ASTM E-729 (1980)
Deviations:
yes
Remarks:
but in various dilution waters
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
no
Test organisms (species):
Oncorhynchus kisutch
Details on test organisms:
TEST ORGANISM
- Common name: Coho salmon
- Source: eye eggs were obtained from Merced River Fish Facility, Snelling, CA, in 1986


ACCLIMATION
- Type and amount of food: Fish were fed a commercial salmon diet

Test type:
static
Water media type:
brackish water
Limit test:
no
Total exposure duration:
96 h
Hardness:
333 +- 9 mg CaCO3/L
Test temperature:
12 +- 1 °C
pH:
7.79+-0.16
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 19.6 L glass jars containing 15 L of test solution
- Aeration:
- No. of organisms per vessel: 10 or 2 x 5 for large, advanced fry
- Biomass loading rate: < 0.8 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The various water types were reconstituted in reverse osmosis-deionized water by varying the type and quantity of mineral salts added. The mineral salts used to reconstitute the various water qualities were calcium chloride, calcium sulfate, magnesium sulfate, sodium bicarbonate, sodium chloride, sodium sulfate and Instant Ocean sea salt. Water quality characteristics were measured according to methods recommended by the APHA (1976) and USEPA (1979).
- Alkalinity: 36.6 +- 1.0 mg CaCO3/L
- Ca/mg ratio: 37.4/58.4 mg/L
- Conductivity: 2887 +- 88 µmhos/cm @ 25 °C


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations on mortality and abnormal behavioral responses were made at 24 h intervals and all dead fish removed after each observation.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
600 mg/L
Nominal / measured:
nominal
Conc. based on:
element
Basis for effect:
mortality (fish)
Reported statistics and error estimates:
The method of Litchfield and Wilcoxon (1949) was used to calculate 96 h LC50 values, 95% confidence intervals, slope function values, and to test for parallelism of concentration mortality curves. The Standard Error of the Difference as described by Sprague and Fogels (1977) and Zar (1974) was used to determine statistical differences in LC50 values.
Sublethal observations / clinical signs:

LC50 after 24 h was >1000 mg/L

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study; well performed study
Reason / purpose for cross-reference:
reference to other study
Qualifier:
according to guideline
Guideline:
other: ASTM E729-95 Standard Guide for Conducting Acute Toxicity Tests on Test Materials with Fishes, Macroinvertebrates, and Amphibians
Qualifier:
equivalent or similar to guideline
Guideline:
EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)
GLP compliance:
no
Remarks:
study is not in compliance with GLP
Analytical monitoring:
yes
Details on sampling:
A sample of each treatment was taken at the beginning and end of the test.
Vehicle:
no
Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: Fathead minnow
- Source: Aquatic Biosystems, Fort Collins Colorado
- Age at study initiation (mean and range, SD): less than 7 d
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
91
Test temperature:
24.7 °C
pH:
8.0
Dissolved oxygen:
> 6.0 mg/L
Nominal and measured concentrations:
Nominal: Control, 15.6, 31.3, 62.5, 125, 250 mg B/L
Measured: <0.022, 15, 32, 64, 125, 255 mg B/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 ml beakers
- Type : open
- Material, size, headspace, fill volume: glass, 200 ml
- Aeration: none
- Renewal rate of test solution: none

- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates):4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: MHRW (moderately hard reconstituted water) per US EPA 2002
- Total organic carbon: 0 nominal
- Chlorine: 0 nominal
- Ca/mg ratio: 1.46
- Culture medium different from test medium: Yes. Test organisms were cultured at Aquatic Biosystems Lab water, hardness 114 mg/L, alkalinity 115 mg/L
- Intervals of water quality measurement: boron, pH, conductivity, alkalinity, hardness at beginning and end of test, temperature and dissolved oxygen daily

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: 16 h light : 8 h dark
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
79.7 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
element
Basis for effect:
mortality (fish)
Remarks on result:
other: (95% CI 72-88 mg B/L), control mortality 2.5%
Sublethal observations / clinical signs:

Biological results:

Measured Concentration (mg B/L) Number dead (out of 40) Mortality [%]
<0.022 (control) 1 2.5
15 3 of 39 7.7
32 0 0.0
64 8 20.0
125 38 95.0
255 40 100.0
Validity criteria fulfilled:
not specified
Conclusions:
The study was conducted according to ASTM E729-95 and EPA OPPTS 850.1075 on the registered substance itself. The method is to be considered scientifically reasonable and the results can be considered as reliable to assess the toxicity of boric acid towards freshwater fish.
The toxic effect of the test item boric acid was assessed on the freshwater species Fathead minnow (Pimephales promelas) in a static 96 h concentration-response test. The 96-hour LC50 was determined to be 79.7 mg B/L. The exposure concentrations were verified by analytical measurements for each treatment at the start and the end of the test. Reported results refer to geometric mean measured concentrations.
Executive summary:

The study was conducted according to ASTM E729-95 and EPA OPPTS 850.1075 on the registered substance itself.

The purpose of this study was to determine the toxicity of the test item to fish. For this purpose, the freshwater species Fathead minnow (Pimephales promelas) was exposed in a static test to various concentrations under defined conditions for 96 hours. The study encompassed 6 treatment groups (5 dose rates of the test item and a control) each containing 40 individuals. The exposure concentrations were verified by analytical measurements. The mortality of the fish was determined by visual observation after 24, 48, 72 and 96 hours.

The 96h-LC50 was determined to be 79.7 mg B/L based on geometric mean measured concentrations.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
disregarded due to major methodological deficiencies
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Guideline study but non-standard species; nominal toxiciyy values: not reliable
Qualifier:
according to guideline
Guideline:
other: ASTM E-729 (1980)
Deviations:
yes
Remarks:
use of various dilution waters
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
no
Test organisms (species):
Oncorhynchus kisutch
Details on test organisms:
- Common name: Chinook salmon
- Source: Fall-run chinook salmon were obtained as eyed eggs from Merced River Fish Facility, Snelling, CA, in 1986.
- Method of breeding: Following hatching, swim-up fry and older fish were cultured according to procedures described by Brauhn and Schoettger (1975).

ACCLIMATION
- Type and amount of food: Fish were fed a commercial salmon diet (Biodiet®).


Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
211 +- 1 mg CaCO3/L
Test temperature:
12 +- 1 °C
pH:
7.82+/-0.35
Nominal and measured concentrations:
Nominal concentrations were expressed as the total element added as determined from the certificate of analysis.
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 19.6 L glass jars containing 15 L of test solution
- No. of organisms per vessel: 10 or 2 x 5 for large, advanced fry
- Biomass loading rate: < 0.8 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The various water types were reconstituted in reverse osmosis-deionized water by varying the type and quantity of mineral salts added. The mineral salts used to reconstitute the various water qualities were calcium chloride, calcium sulfate, magnesium sulfate, sodium bicarbonate, sodium chloride, sodium sulfate and Instant Ocean sea salt. Water quality characteristics were measured according to methods recommended by the APHA (1976) and USEPA (1979).
- Alkalinity: 87.2 +- 0.8 mg CaCO3/L
- Ca/mg ratio: 45.8/23.6 mg/L
- Conductivity:721 +- µmhos/cm @ 25 °C


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations on mortality and abnormal behavioral responses were made at 24 h intervals and all dead fish removed after each observation.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
447 mg/L
Nominal / measured:
nominal
Conc. based on:
element
Basis for effect:
mortality (fish)
Remarks on result:
other: CI = 356 - 561 mg/L
Reported statistics and error estimates:
The method of Litchfield and Wilcoxon (1949) was used to calculate 96 h LC50 values, 95% confidence intervals, slope function values, and to test for parallelism of concentration mortality curves. The Standard Error of the Difference as described by Sprague and Fogels (1977) and Zar (1974) was used to determine statistical differences in LC50 values.
Sublethal observations / clinical signs:

LC50 after 24 h was >1000 mg/L

Description of key information

Only 1 short-term toxicity value for freshwater fish has been selected, i.e. for the fathead minnow P. promelas with a 4 days LC50 of 79.7 mg B/L. 
Only 1 short-term toxicity value for marine fish has been selected, i.e. for the dab L. limanda with a 4 days LC50 of 74.0 mg B/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
79.7 mg/L

Marine water fish

Marine water fish
Effect concentration:
74 mg/L

Additional information