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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:
key study
Study period:
1979-07-1981-12
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study was conducted in conformance to GLP and in accordance to ASTM guideline. Chemical analyses of test solutions were absent in the study design, however the data of section 3.1.2 (Stability in water) indicate sufficient exposure. Dissolved oxygen concentration in test system fell below 40% in the exposure scenario with the subadult fish, therefore toxicity data from this particular exposure was not used as the effects of low oxygen concentrations could not be separated from the inherent toxicity of the test material.
Qualifier:
according to guideline
Guideline:
other: ASTM Standard E 729-80. Standard practice for conducting acute toxicity tests with fishes, macroinvertebrates and amphibians. Philadelphia. Penna: American Society for Testing and Materials; 1980
Deviations:
yes
Remarks:
The dissolved oxygen concentration fell below the recommended limit of 40% saturation, but only in the exposure scenario with subadult fish.
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
not specified
Details on test solutions:
The dilution water supply for the laboratory was from the upper Saginaw Bay of Lake Huron. Water was carbon filtered and U.V. irradiated prior to use. During the course of this study the pH ranged from 7.6 to 8.3; hardness ranged from 96 to 125 (mg /~as CaC03) and the conductivity ranged from 135-215 ~mhos/cm.



Each test consisted of at least six exposure concentrations in addition to a water control and an acetone control (if this carrier was used). Exposure was initiated by thoroughly mixing a measured volume of a stock solution to 2 liters of dilution water (500 mL with smaller test vessels) and adding the mixture to the appropriate test vessel . The final volume in the large vessels was 10 liters; and 3.5 liters in the small vessels. Volatile compounds were tested in vessels either covered with SARAN WRAP@ plastic film or screw-top lids to retard volatilization.
Test organisms (species):
Pimephales promelas
Details on test organisms:
- Age/size/weight/loading:
age (days) size (mm) weight (mg)
fry 10-15 9.5 11.6
juvenile 30-35 14.9 76.8
subadult 65-94 28.0 391
- Feeding: Newly hatched fry were fed with Artemia salina (2
times per hour), older fish were fed daily with a synthetic
diet
- Pretreatment: All fish were held in 57 liter aquaria with a
constant water flow of 0.2 l/minute at a temperature of 25
± 1°C, illumination of 430-645 lux and a light-dark cycle
of 16h:8h. Fish were accclimated to the test temperature of
22 ± 1°C at least 48 hours prior to the start of the test
- Feeding during test: no
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
Study was terminated after 96 hours.
Hardness:
96-125 mg/l as CaCO3
Test temperature:
Exposure with Fry: 21.2 - 21.8 C
Exposure with Juveniles: 21.3 - 22.2 C
Exposure with Subadults: 21.0 - 22.0 C
pH:
Exposure with Fry: 7.6 - 8.4
Exposure with Juveniles: 7.5 - 8.3
Exposure with Subadults: 7.2 - 8.1
Dissolved oxygen:
Exposure with Fry: 6.5 - 8.7 mg/L
Exposure with Juveniles: 7.2 - 8.2 mg/L
Exposure with Subadults: 2.2 - 6.6 mg/L
Salinity:
no data
Nominal and measured concentrations:
6 exposure concentrations and one control
Details on test conditions:
Photoperiod: 16h:8h light-dark cycle

TEST SYSTEM
- Test type: static with aeration and without renewal of test solution
- Concentrations: six exposure concentrations and one control
- Exposure vessel type: round glass vessels measuring 22 cm deep and 24.5 cm in diameter containing 10 liters (juvenile and subadult test) or 3.5 liters (fry test)
- Number of replicates, fish per replicate: Number of replicates is not described, 10 subadults, 10 juveniles or 20 fry

DILUTION WATER
- Source: Lake Huron water, carbon filtered and UV irradiated
- Alkalinity: not described
- Hardness: 96-125 mg/l as CaCO3
- pH: 7.9-8.3
- Conductance: 135-215 µmhos/cm

Three age groups of fish were tested and were defined as follows: fry (10-15 day), juveniles (30-35 day) and subadults (60-100 day). Tests with the subadults and juveniles were conducted in round glass vessels measuring 22 cm deep and 24.5 cm in diameter. Fry tests were conducted in the above described vessels, 4 liter beakers (25 cm x 15 cm) or screw-top gallon jars (24 cm x 14 cm). Constant temperature water troughs were set to maintain the test temperature at 22 +/- 1°C. Twenty-four hours prior to the beginning of a test, 10 subadults, 10 juveniles or at least 20 fry were indiscriminately placed in each test and control vessel containing either 8 liters or 3 liters (fry tests) of 22C dilution water. The aquaria were gently aerated during the 24 hour acclimation period. The subadults and juveniles were not fed during the 24 hours preceding the test, nor during the testing period. Fry were fed prior to, but not during the test.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
12.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 11.5 -14.2 mg/L; Fry
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
10.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 9.1 - 12.3 mg/L; Juvenile
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
13.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
However, low DO levels compromise this data point
Basis for effect:
mortality (fish)
Remarks on result:
other: 9.6 - 18.6 mg/L; Subadult
Details on results:
The following LC50 (96h) with 95% Confidence Intervals were found:
- fry (10-15 day) : 12.7 mg/l (11.5-14.2 mg/l)
- juvenile (30-35 day) : 10.6 mg/l (9.1-12.3 mg/l)
- subadult (65-94 day) : 13.2 mg/l (9.6-18.6 mg/l)
Results with reference substance (positive control):
no data
Reported statistics and error estimates:
not applicable
Sublethal observations / clinical signs:

The dissolved oxygen concentration fell below the recommended limit of 40% saturation in the exposure test with subadult fish. Therefore,the LC50 data point from this exposure scenario is not wholly reliable since the stressful effects of low oxygen levels can not be separated from the inherent toxicity of the test substance.

There were no apparent trends in the sensitivity of a particular age group to the compounds tested .

Conclusions:
96 hr LC50 value for epichlorohydrin is: Fry - 12.7 mg/L (11.5 - 14.2 mg/L)
Juvenile - 10.6 mg/L (9.1 - 12.3 mg/L)
Subadult - 13.2 mg/L (9.6 - 18.6 mg/L)
Executive summary:

The 96 hour LC50 was determined for epichlorohydrin in fry, juvenile and subadult Pimephales promelas. There was little difference between the three age groups with calculated 96 hour LC50 values of 12.7 mg/L for fry, 10.6 mg/L for juveniles and 13.2 mg/L for subadults.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study is relatively well-documented, however there were no analyses of test solutions concentrations in this study.
Qualifier:
no guideline available
Principles of method if other than guideline:
All bioassays were conducted at the Unites States Testing Company Incorporated, Biological Services Division, Hoboken, New Jersey, USA
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
no data
Analytical monitoring:
no
Details on sampling:
no data
Vehicle:
not specified
Details on test solutions:
Potable well water, pH 7.6 -7.9 and hardness 55 mg/L CaCO3, was used as the base for a synthetic seawater mix. "Instant Ocean" synthetic sea salt mix was added to freshwater until a specific gravity of 1.018 was achieved. The salt concentration corresponded with the specific gravity of the natural seawater the specimens were collected from. The necessary volume of altwater for testing was made one day in advance and stored in clean polyethylene vessels at a uniform temperature.
Test organisms (species):
Menidia beryllina
Details on test organisms:
Tidewater silversides (Mendidia beryllina) were collected in nets primarily from "Horseshoe Bay" at Sandy Hook, New Jersey, USA (Gateway National Park). The fish were held for an acclimation period of fourteen days prior to testing. As in the freshwater acclimation, any group showing greater than 5 percent mortality was judged "unfit". Fish were maintained exclusively on a minced frozen shrimp diet.

Fish approximately 40 mm to 100 mm were selected at random for the assays. Only fish from healthy tanks were used.
Test type:
static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
The test was terminated after 96 hours.
Hardness:
No data
Test temperature:
Facilities were housed in an air conditioned room with constant temperature setting of 20 C.
pH:
No data
Dissolved oxygen:
No data
Salinity:
"Instant Ocean" synthetic sea salt mix was added to freshwater until a specific gravity of 1.018 was achieved, This salt concentration corresponded with the specific gravity of the natural seawater the specimens were collected from.
Nominal and measured concentrations:
Nominal: control, 10, 18, 32 mg/L
Details on test conditions:
All glass 5-gallon aquaria were used as test trial containers. One gallon glass wide mouth jars (without lids) were used as screening containers. Water levels in the containers were brought to a depth of greater than 15 cm for testing purposes and the total volume adjusted to assure that a minimum of 1liter of water was present for every 1 gram of fish. Test containers were places on racks mounted with individual valve-operated aeration setups which provided oil-free compressed air if necessary.

Mortality counts were taken daily and dead fish were immediately removed at the time of first observation.
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
18 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
no data
Results with reference substance (positive control):
no data
Reported statistics and error estimates:
no data
Validity criteria fulfilled:
not applicable
Remarks:
The estimated death rate of control fish during the experiment was about 3.0 percent.
Conclusions:
The 96-hour LC50 for Menidia beryllina exposed to epichlorohydrin was 18 mg/L.
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1977
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study is relatively well-documented, however there were no analyses of test solutions concentrations in this study.
Qualifier:
no guideline available
Principles of method if other than guideline:
All bioassays were conducted at the Unites States Testing Company Incorporated, Biological Services Division, Hoboken, New Jersey, USA.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
no data
Analytical monitoring:
no
Details on sampling:
no data
Vehicle:
no
Details on test solutions:
Potable well water obtained from an underground source in Passaic County was used as the holding and dilution water for the testing. The pH of the water was 7.6 - 7.9 with a hardness of 55 mg/L CaCO3. The water was collected on a weekly basis and stores in clean polyethylene vessels at a uniform temperature.

Dilutions of the test substances, when necessary, were made in distilled water or in a solvent with relatively low toxicity.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
Bluegill sunfish obtained from commercial hatcheries in the vicinity of the New York metropolitan area were used. The fish were held for an acclimation period of fourteen days prior to testing. During that time, holding tank counts were taken and any group showing greater than 5 percent mortality was judged unfit. Fish were maintained in a commercial fish diet supplemented with minced frozen shrimp.

Fish, approximately 33 mm to 75 mm in length, were selected at random for the assays. Fish were not fed for 48 hours prior to testing. The trial fish were placed in the assay vessels before doses of the chemcials were added.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
The test was terminated after 96 hours.
Hardness:
Dilution water was reported to have a hardness of 55 mg/L CaCO3.
Test temperature:
Facilities were housed in an air conditioned room with constant temperature settinf of 23 degrees C.
pH:
pH was noted at the end of the assay time period.
Dissolved oxygen:
Dissolved oxygen readings were taken daily. If dissolved oxygen was being depleted rapidly, either by the test organism or chemical and biochemical demand, aeration was initiated.
Salinity:
Not applicalbe
Nominal and measured concentrations:
Nominal concentrations: control, 10, 32, 37, 42, 56 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: all glass 5-gallon aquaria
- Material, size, headspace, fill volume: water levels in the containers were brought to a depth of greater than 15 cm and the total volume adjusted to assure that a minimum of 1 liter of water was present for every gram of fish
- Aeration: yes, if necessary
- Biomass loading rate: 1 g/L



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality counts were taken daily.


Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
35 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
no data
Results with reference substance (positive control):
no data
Reported statistics and error estimates:
no data
Sublethal observations / clinical signs:

no data

Validity criteria fulfilled:
not applicable
Remarks:
The estimated death rate of control fish during the experiment was about 1.3 percent for freshwater fish.
Conclusions:
The 96 hour LC50 for epichlorohydrin with the bluegill sunfish is 35 mg/L.

Description of key information

The 96-hour LC50 value of 10.6 mg/L for epichlorohydrin is taken from a valid exposure study with fathead minnows (Pimephales promelas).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
10.6 mg/L

Additional information

Two acute freshwater fish studies and one acute saltwater fish study were assessed for this endpoint. One study was conducted with fathead minnows (Pimephales promelas), one study was conducted with bluegill sunfish (Lepomis macrochirus) and one study was conducted with tidewater silversides (Menidia beryllina). All three studies were determined to be of good quality and acceptable for use in risk assessment. The study conducted with fathead minnows tested the sensitivity of fish at different life-stages to epichlorohydrin, and concluded that there was no real difference in sensitivity to epichlorohydrin based on age of the exposed fish. The LC50 values for fathead minnow fry, juveniles, and subadults were 12.7, 10.6 and 13.2 mg/L epichlorohydrin, respectively. The acute test with epichlorohydrin and bluegill sunfish reported a 96-hour LC50 of 35 mg/L. The acute test with epichlorhydrin and tidewater silversides (a saltwater species) reported a 96-hour LC50 of 18 mg/L. The lowest 96-hour LC50 value of 10.6 mg/L for fathead minnows was selected as the key parameter for this endpoint.