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EC number: 213-690-5 | CAS number: 1002-67-1
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Long-term toxicity to fish
Administrative data
Link to relevant study record(s)
- Endpoint:
- fish early-life stage toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 12/11/2021 - 16/12/2021
- Reliability:
- 1 (reliable without restriction)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)
- Deviations:
- yes
- Remarks:
- See "any other information on materials" for details
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- Test Item name 1-ethoxy-2-(2-methoxyethoxy)ethane
Lot number 09511
Active ingredient content: ˃ 99.9%
Appearance water clear, liquid
Expiry date March 28, 2023
Storage at room temperature, protect from light - Analytical monitoring:
- yes
- Remarks:
- Analytical control of the test solutions was performed with a validated method (Validation Study No.: 103-100-6044) by the Analytical Laboratory of TOXI-COOP ZRT. The samples were analysed GC-MS method.
- Details on sampling:
- The selected nominal concentrations (spaced by a
factor of 2.3) were 0.36 mg/L, 0.82 mg/L, 1.9 mg/L, 4.3 mg/L and
10 mg/L which are the same as was used in the Daphnia magna 21-day
reproduction test performed previously in the testing laboratory
(Study No.: 103-211-6041).
For determination of 1-ethoxy-2-(2-methoxyethoxy)ethane
concentration samples were taken from the test solutions and from the
negative control at the start and at the end of the renewal periods. Test
item concentrations were analysed in the test solutions by a GC-MS
method. 1-ethoxy-2-(2-methoxyethoxy)ethane concentration was
determined at 9 renewal periods.
During the test, the concentration of the test item in the test solutions were determined at
regular intervals to characterise exposure. Both the freshly prepared test solutions (“new”
test solutions) and the test solutions removed from the test chambers (“old” test solutions)
before use of the “new” solutions were analysed regularly.
Determinations were made for 9 renewal periods. Both “new” and “old” test solutions were
analysed in all separate test chambers at each occasion.
Samples were taken on the following days: 0, 4, 7, 11, 14, 18, 21, 25, 31 and 34. - Vehicle:
- not specified
- Details on test solutions:
- The test solutions used in the test were prepared by mechanical dispersion without using of any solubilising agent, as the test item was well water soluble. A stock solution was first prepared by dissolving an appropriate amount of test item in the dilution water, then the test solutions were prepared by appropriate dilutions of this stock solution. Once the test solutions prepared the appropriate volume was used for transfer of the test animals and the remaining test solutions were stored under ambient temperature (at the same place as the test aquariums) and used as necessary for renewal of the test solutions in the test aquariums (by replenishment or transfer,)
Control
A dilution water control was run in addition to the test item concentration series.
Table 3. Preparation of the Stock and Test Solutions see "any other information on results". - Test organisms (species):
- Danio rerio (previous name: Brachydanio rerio)
- Details on test organisms:
- Species: Zebrafish (Danio rerio)
Origin: The breeding stocks used for production of eggs is obtained from
Toma István e.v.
Hungary, 2146 Mogyoród, Gödöllői u. 41/B. and
from Hungarian University of Agriculture and Life Sciences Hungary, 2103 Gödöllő, Páter K. u. 1.
Number of eggs / test groups: 100 (divided equally between the replicates)
Number of replicates / test groups: 4
Stage of eggs at start of exposition: The embryonic stage at the beginning of exposure was verified as much as possible: stage of embryonic development at the time of pre-treatment was approximately 2-32 cells in the test solutions and the control. - Test type:
- semi-static
- Water media type:
- other: Reconstituted water diluted with purified water in a ratio of 3 : 2 (3 part reconstituted water: 2 part purified water) was used as dilution water in the test. see "any other information on results"
- Limit test:
- no
- Total exposure duration:
- 34 d
- Hardness:
- see "any other information on results"
- Test temperature:
- 26 ± 1.5 °C
see "any other information on results" - pH:
- see "any other information on results"
- Dissolved oxygen:
- 60.7 % – 95.4 % air saturation value
see "any other information on results"
Test solutions were slightly aerated from Day 5 to Day 24 (during the period when dry food was provided) - Salinity:
- not specified in study report
- Conductivity:
- not specified in study report
- Nominal and measured concentrations:
- see Table 8 Summarized Results of Analytical Determination of the Test Concentrations (Determined for 9 Renewal Periods) in "any other information on results"
- Details on test conditions:
- Test Chambers
Type and Size: Suitable sized glass Petri-dishes with 25-30 mL test solutions (until free-feeding stage);
Suitable sized glass aquaria with 0.25-0.5 L test solutions (from free-feeding stage to the end of the test).
Identification: Each test unit was uniquely identified with study number, test item concentration and number of parallels.
Allocation: Until free-feeding stage the test chambers were kept in a temperature-controlled incubator. The free-feeding larvae were transferred to suitable sized test aquariums and kept on a temperature-controlled tray until end of the test.
Environmental Conditions
Temperature: The temperature was maintained within a range of 26 ± 1.5 °C and was appropriately constant (the water temperature could not differ by more than ± 1.5 °C between the test chambers or between the successive days at any time during the test).
Photoperiod: 16 h light: 8 h dark
Oxygen concentration: Test solutions were not aerated until start of feeding (from Day 0 to Day 4). After start of feeding of the larvae, test solutions were slightly aerated during the period when dry food was given (from Day 5 to Day 24). No aeration was used from Day 25 to Day 34.
Duration of the Test
The test started as soon as it was technically possible after the eggs have been fertilized and was continued 30 days post-hatch of the control fish.
Replenishment and Renewal of the Test Solutions, Transfer of the Test Organisms
Renewal of the test solutions was performed by transfer of the test organisms from “old” test solutions to “new” test solutions. Replenishment of the test solutions (lost because of removal of uneaten food and faces) was also performed.
For renewal a new batch of test solutions were prepared. Within the stability period (i. e. 7 days; determined during the analytical method validation) these test solutions were used as necessary for replenishment or for transfer of the test organisms.
Days of preparation of a new batches of test solutions: Days 0, 4, 7, 11, 14, 18, 21, 25 and 31.
Days of transfer of test animals: from Day 4 to Day 25 and on Day 31 (fish were transferred in the afternoon before feeding)
Days of replenishment of the test solutions (without transfer of the test animals): - once a day at the observation: from Day 0 to Day 3
- once a day in the morning, before the feeding: from Day 5 to Day 25 and on Day 31 - twice a day before the feedings: from Day 26 to Day 30 and on Days 32, 33.
Loading
One hundred eggs, divided equally between the four replicates, were used per test concentrations and the control. Volume of the test solutions was increased from 25-30 mL to 250 mL (on Day 4) and finally to 500 mL (from Day 5 to Day 34) to maintain loading of the test chambers as low as technically possible during the test.
Feeding
The fish were fed from elimination of yolk sac (free-feeding stage) to the end of the test twice a day with appropriate type and amount of food (except on the first feeding day when food was supplied once only).
Dry food (fine powder) was introduced in the afternoon of Day 4. Live food (Artemia naupli; AN) was introduced on Day 14. From Day 14 to Day 24 both type of food (DF and AN) was supplied to the animals. Fish were not fed on the day of termination of the test (Day 34). Both type of food was supplied ad libitum (means supplying excessive amount of food for the test organisms regularly).
Dry food (DF) was weighed and suspended in the dilution water at each occasion.
On Day 4 0.35 g dry food was suspended in 14 ml dilution water and same volume (0.5 mL) of this suspension was supplied for each test aquarium (in the afternoon only) meaning 12.5 mg dry food/test aquarium/day.
From Day 5 to Day 20 0.35 g dry food was suspended in 14 ml dilution water and same volume (0.5 mL) of this suspension was supplied for each test aquarium twice a day meaning 25.0 mg dry food/test aquarium/day.
From Day 21 to Day 24 0.175 g dry food was suspended in 7 mL dilution water and 0.25 mL of this suspension was supplied for each test aquarium twice a day meaning
12.5 mg dry food/test aquarium/day.
For producing live food (AN) brine shrimp eggs (approximately 1.5 mL) were placed into 500 mL salt water (6.25 g or 10 g NaCl/500 mL dilution water) and left to hatch for approximately 24 or 36 hours. Two parallels were prepared in the morning of each day. Hatched nauplies (one portion in the morning and one in the afternoon) were filtered and suspended in 14 mL dilution water and 0.5 mL of this suspension was supplied for each test aquarium twice a day from Day 14 to Day 33.
Dry food used in the test:
Name: Sera Micron Nature Appearance: Fine powder
Main components: Spirulina (51 %), krill (18 %), Artemia Shrimp, Omega fatty acids
(34%), fish flour, wheat flour, Brewer’s yeast
Manufacturer: Sera, D-52518
Heinsberg, Germany
Storage: 2-8 °C
Identification Code: SM210712/1 and SM210712/2
Charge No.: 0000077382 Expiry date: 01/2024
Live food (Artemia nauplii) prepared from:
Name: Artemio®Pur
Artemia (brine shrimp) eggs
Manufacturer: JBL GmbH & Co. KG, D-67141
Neuhofen, Germany
Storage: 2-8 °C
Identification Code: AP210712/1 and AP210712/2
Expiry date: 03/2023
Feeding regime is summarized in Table 24 see "any other information on results".
Cleaning
Surplus food and faces were removed by syphoning before replenishment of the test solutions (in the mornings) and by transfer of the test animals into a clean test aquarium (in the afternoons).
Water Quality Measurements
The water temperature was determined at least once a day in all test chambers. In addition, water temperature was continuously monitored with a min/max thermometer in a concurrent test chamber containing the dilution water only at the same volume as the test solutions in the test chambers. Measured temperature values are given in Table 16 and Table 17 see "any other information on results" .
Dissolved oxygen concentrations (LDO) and pH were determined at least weekly in all test chambers.
LDO was measured on Days 0, 4, 5, 6, 7, 8, 11, 12, 18, 19, 21, 22, 25, 26, 29, 30, 33 and 34. Measured values are given in Table 18 see "any other information on results" . pH was measured on Days 0, 4, 5, 11, 12, 18, 19, 25, 26, 29 and 34. Measured values are given in Table 19 "any other information on results"
Total hardness (mg/L CaCO3) was measured at the beginning of the test (on Day 0) in each test solutions (including control) and at the end of the test (on Day 34) in all test aquariums (see "any other information on results").
Recording
Test conditions were measured with suitable instruments and documented in the raw data.
- Reference substance (positive control):
- not specified
- Key result
- Duration:
- 34 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- weight
- Key result
- Duration:
- 34 d
- Dose descriptor:
- LOEC
- Effect conc.:
- > 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- weight
- Key result
- Duration:
- 34 d
- Dose descriptor:
- LOEC
- Effect conc.:
- > 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- length
- Key result
- Duration:
- 34 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- length
- Key result
- Duration:
- 34 d
- Dose descriptor:
- LOEC
- Effect conc.:
- > 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Key result
- Duration:
- 34 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Key result
- Duration:
- 34 d
- Dose descriptor:
- LOEC
- Effect conc.:
- > 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- number hatched
- Key result
- Duration:
- 34 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- number hatched
- Details on results:
- Environmental Conditions
Temperature: The temperature was maintained within a range of 26 ± 1.5 °C and was appropriately constant. The overall temperature range in the test chambers was 25.1 - 26.5 °C with a maximum difference of 1.4 °C. The measured water temperature range in a concurrent test chamber (monitored continuously with a min/max thermometer) was 25.1 - 26.5 °C with a maximum difference of 1.4 °C. Results are summarized in Table 4 see "any other information on results". Measured temperature values are given in Table 16 and Table 17 see "any other information on results".
Oxygen concentration: Values over 60 % of the air saturation value (ASV) were observed during the test. The overall range in the test chambers was 60.7 % – 95.4 % ASV. Test solutions were slightly aerated from Day 5 to Day 24 (during the period when dry food was provided). Results are summarized in Table 5. Measured values are given in Table 18 see "any other information on results".
pH: Summarized results of the measured pH values are given Table 6 see "any other information on results". Measured values are given in Table 19 see "any other information on results".
Water Hardness: Water hardness was measured at the beginning and end of the test. Measured values are given in Table 7 see "any other information on results".
Analytical Results
1-ethoxy-2-(2-methoxyethoxy)ethane concentration in the test solutions was determined at 9 renewal periods. Five different concentrations and the control group were tested. The nominal test concentrations were 0.36 mg/L, 0.82 mg/L, 1.9 mg/L, 4.3 mg/L and 10 mg/L. Four replicates were tested per each test group. Samples were taken and analysed by GC-MS method. The analytical part of the study (method, determination, results) is attached see "any other information on results". The results of analytical determination of the test concentrations are given in Table 8 see "any other information on results". .
Biological Results
Since the analytically measured test item concentrations were within the range of ± 20 % of the nominal values over the test period of 34 days biological results are based on the nominal concentrations.
Survival, hatching, body weight and body length data in the test groups were compared to the control.
Hatching and Survival
Hatching of larvae started on Day 3 in each test chambers. All survival larvae were hatched in the control and also in the test groups on Day 4 (end of hatching period). Observations on hatching and the numbers of larvae hatched daily are given in Table 20 see "any other information on results". .
Test organisms were observed at least once a day during the test and numbers of living animals, the mortalities and numbers of animals with observed abnormalities (body deformities and/or abnormal behaviour) were also recorded. The observations are given in Table 21 see "any other information on results".
Data evaluation on hatching and survival was performed: hatching success, post-hatch success, overall survival and cumulative mortality were calculated. Tabulated results are given in Table 9 see "any other information on results".
Hatching success (%) varied between 72.0 % and 96.0 % in the test chambers. The mean values were within the range of 80.0 % and 91.0 %.
Post-hatch success in the test chambers varied between 70.8 % and 89.5 %. The mean values were within the range of 75.9 % and 80.2 %.
The following endpoints were statistically evaluated in the test groups in comparison with the control data:
- hatching on Day 3 and Day 4 (see Table 10 "any other information on results". )
- post-hatch survival for period Day 4 – 14 (feeding with dry food only; see Table 11 "any other information on results". )
- post-hatch survival for period Day 4 – 25 (feeding both with dry food and live food; see Table 12 "any other information on results". );
- post-hatch survival for the whole juvenile stage (Day 4 - Day 34, see Table 13 "any other information on results". ).
No statistically significant effect on the hatching was observed on Day 3 or on Day 4 at any test concentrations.
No statistical significance was observed for post-hatch survival during any stage of the test at any concentration.
Based on this it is considered that the test item did not influence the survival or hatchability of the test organism.
Body Weights and Body Lengths
At the end of the test all surviving fish were weighed. Total body weights were measured on a replicate basis due to small size of the fish (see Table 22 "any other information on results". ). Mean body weights calculated for the surviving individual fish were statistically evaluated. No statistically significant differences were observed (see Table 14 "any other information on results". ). No dose-related changes were observed between the test item treated groups.
Body length was measured for each individual fish (see Table 23 see "any other information on results". ). Mean body length per surviving individuals were statistically evaluated (see Table 15 "any other information on results". ). No statistically significant differences were observed. No dose-related changes were observed between the test item treated groups.
Symptoms of intoxication were recorded during the test with limited extent. Numbers of abnormal animals observed during the test are given in Table 21 see "any other information on results".
Although data were not statistically evaluated, no obvious treatment-related symptoms were considered. The observed abnormalities were mainly gross body deformities. The nature and frequency of the observed abnormalities in the test item treated groups did not differ from the control and did not show any dose dependence. Neither of the animals with observed abnormalities survived the test, since no fish were observed with abnormal appearance or behaviour at the end of the test.
Based on the overall results the following biological endpoints can be determined for 1-ethoxy-2-(2-methoxyethoxy)ethane. - Results with reference substance (positive control):
- not specified in the study report.
- Reported statistics and error estimates:
- The statistical analysis was performed by ToxRat Professional (Version 3.3.0) software.
To determine NOEC/LOEC values for hatchability and post-hatch survival, Chi2 test with Bonferroni Correction was performed.
To determine NOEC/LOEC values for body weight and body length, Dunnett’s Multiple t-test (at the 0.05 significance level) was performed.
See "illustration". - = before renewal of the test solution
- = after renewal of the test solution
- = before renewal of the test solution
- = after renewal of the test solution
- = before renewal of the test solution
- = after renewal of the test solution
- 0 89.9 72.8 90.2 74.0 90.4 75.0 93.1
- 688.1 89.7 63.0 95.4 74.1 89.7 71.9 90.4 73.1 89.8 75.4 92.5 74.1 89.9 74.2 93.0 76.1 70.6 69.4 68.9 71.2
- 688.1 90.5 64.6 94.2 73.1 91.2 70.0 89.9 71.0 90.0 73.0 92.7 74.2 91.5 74.8 91.8 74.9 69.8 69.9 71.5 69.9
- 688.1 89.9 62.7 94.1 70.8 90.4 69.9 93.1 71.6 91.6 72.9 90.6 71.6 90.6 72.6 91.5 74.6 71.1 71.0 70.9 68.9
- = = measured in the prepared test solutions used for treatments a* = measured in the old test solution pooled according to treatment group (at transfer of the animals) a = measured in the clean solution after the transfer b = measured in the old solution before the transfer
- = = measured in the prepared test solutions used for treatments a* = measured in the old test solution pooled according to treatment group (at transfer of the animals) a = measured in the clean solution after the transfer b = measured in the old solution before the transfer
- Validity criteria fulfilled:
- yes
- Remarks:
- The water temperature met with the validity criteria. The dissolved oxygen concentration in the test solutions also met with the criteria. According to this the assay is considered valid and the obtained results are reliable.
- Conclusions:
- Under the conditions of this fish early life stage toxicity test, the obtained results showed that the test item 1-ethoxy-2-(2methoxyethoxy)ethane had no significant lethal or sub-lethal effects on early life stages of Zebrafish (Danio rerio) up to at least the highest tested concentration of 10 mg/L. Accordingly, the test item 1-ethoxy-2-(2-methoxyethoxy)ethane had no chronic effects on Zebrafish (Danio rerio) up to at least the tested maximum concentration of 10 mg/L.
- Executive summary:
Test Item:
1-ethoxy-2-(2-methoxyethoxy)ethane
Test species:
Zebrafish (Danio rerio)
Purpose:
The purpose of this study was to evaluate the chronic toxicity of the test item 1-ethoxy-2-(2-methoxyethoxy)ethane to early life stages (embryo, larvae and juveniles) of zebrafish (Danio rerio). For this purpose, eggs were exposed in a semi static test to aqueous test media containing the test item for 34 days at a range of concentrations under defined conditions. Lethal and sub-lethal effects were assessed and compared with the solvent control values to determine the lowest observed effect concentration (LOEC) and hence the no observed effect concentration (NOEC).
Dilution water:
Reconstituted water (ISO medium) diluted with purified water (in a ratio of 3 : 2 = reconstituted water: purified water)
Control:
Negative control (dilution water)
Test Item Concentrations, Formulation:
Test concentrations were selected according to the results of the preliminary test (acute 96 hours fish test on Zebrafish) and in line of the OECD 210 guideline. The selected nominal concentrations (spaced by a factor of 2.3) were 0.36 mg/L, 0.82 mg/L, 1.9 mg/L, 4.3 mg/L and 10 mg/L which are the same as was used in the Daphnia magna 21-day reproduction test performed previously in the testing laboratory (Study No.: 103-211-6041).
Analytics:
For determination of 1-ethoxy-2-(2-methoxyethoxy)ethane concentration samples were taken from the test solutions and from the negative control at the start and at the end of the renewal periods. Test item concentrations were analysed in the test solutions by a GC-MS method. 1-ethoxy-2-(2-methoxyethoxy)ethane concentration was determined at 9 renewal periods.
Test design:
One hundred eggs (attributed randomly to the groups and distributed equally between the four replicates) were tested per test concentrations and the control. Based on microscopic observation embryos were at approximately 2 to 32 cell stages at start of the treatment (in the pre-treatment solutions). Test solutions were renewed in each test chamber either with replenishment of the test solutions (in the mornings) or with transfer of the test animals into a clean solution (in the afternoons).
Test organisms were kept in the incubator until Day 4 [until the test animals in the test chambers (glass Petri-dishes) become free-feeding]. Larvae were transferred to the test chambers (glass aquariums) on Day 4 before elimination of yolk sac and feeding was initiated.
Animals were fed from the free-feeding stage to the end of the test with appropriate type and amount of food ad libitum. The same amount of food was supplied for each test chamber twice a day (except on Day 4) in excess at the time of renewal of the test solutions.
Food was added to the test chambers either after replenishment of the test solutions or after transfer of the test animals into a clean test chambers.
Test organisms were observed once a day (mainly at the time of the transfers). At the end of the test animals were euthanized with hypothermic shock.
Measurements:
Environmental conditions (temperature, dissolved oxygen concentration, pH, water hardness) were monitored during the test.
Observations:
Mortality, hatching, body weight, body length, abnormal appearance/behaviour were observed during the test.
Statistics:
Results:
The statistical analysis was performed by ToxRat Professional (Version 3.3.0) software.
To determine NOEC/LOEC values for hatchability and post-hatch survival, Chi2 test with Bonferroni Correction was performed.
To determine NOEC/LOEC values for body weight and body length, Dunnett’s Multiple t-test (at the 0.05 significance level) was performed.
Validity:
The water temperature met with the validity criteria and the test item was analytically determined at regular intervals. Hatching success as well as post-hatch success in the control group were over the specified value of 70 % or 75 % (respectively).
The dissolved oxygen concentration in the test solutions also met with the criteria (the measured values were over the 60 % of the air saturation value during the test).
According to this the assay is considered valid and the obtained results are reliable.
Analytical results:
The analytically measured test item concentrations (determined at 9
renewal periods) were within the range of ± 20 % of the nominal values over the test period of 34 days.
Biological results: Based on the results of the concentration analysis biological results were reported according to the nominal test concentrations which were 0.36 mg/L, 0.82 mg/L, 1.9 mg/L, 4.3 mg/L and 10 mg/L.
Hatching, post-hatch survival, body weight and body length data were evaluated compared to the control.
Number of larvae hatched daily was statistically evaluated on Day 3 and Day 4. No statistical significance was observed for hatching at any test concentrations.
Post-hatch survival was statistically evaluated on Days 14, 25 and 34. No statistical significance was observed for hatching at any test concentrations on any of the days.
Total body weights were measured on a replicate basis. Mean body weights (calculated from the total body weights for the individual fish) were statistically evaluated. No statistically significant differences were observed at any test concentrations.
Body length was measured for each individual fish and the data were
statistically evaluated. No statistically significant differences were observed.
Other sub-lethal effects were also monitored and numbers of embryos/larvae/juveniles with deformities and / or abnormal behaviour were recorded when observed. Although these data were not statistically evaluated, no obvious test item related changes in the appearance or behaviour of the animals were observed.
Table 1. Summary of Biological Endpoints
Endpoints
1-ethoxy-2-(2-methoxyethoxy)ethane concentration
(mg/L)
Hatchability NOEC
≥ 10
Hatchability LOEC
> 10
Post-hatch survival NOEC
≥ 10
Post-hatch survival LOEC
> 10
Body weight (terminal) NOEC
≥ 10
Body weight (terminal) LOEC
> 10
Body length (terminal) NOEC
≥ 10
Body length (terminal) LOEC
> 10
34 day NOEC
≥ 10
34 day LOEC
> 10
34 day ECx
> 10
Conclusion: Under the conditions of this fish early life stage toxicity test, the obtained results showed that the test item 1-ethoxy-2-(2methoxyethoxy)ethane had no significant lethal or sub-lethal effects on early life stages of Zebrafish (Danio rerio) up to at least the highest tested concentration of 10 mg/L. Accordingly, the test item 1-ethoxy-2-(2-methoxyethoxy)ethane had no chronic effects on Zebrafish (Danio rerio) up to at least the tested maximum concentration of 10 mg/L.
Reference
Analytical Investigations:
Measurement of Test Item Concentration in the Test Solutions
Table 1. Other materials
Chemical | Grade | Manufacturer/ Supplier | Batch / Lot Number | Expiry/Retest date |
Acetonitrile | for HPLC super gradient | VWR | 19L021843 | December 02, 2022 |
Ultra-pure water | ASTM Type I | prepared by Synergy smart UV HF, MILLIPORE |
Table 2. Results of the method validation (Study No.: 103-100-6044)
Specificity | Interference was less than 20 % of the lowest concentration of the calibration curve |
Repeatability/Instrument precision (7 injections) | 1.8% (0.1 µg/mL) and 0.2% (8 µg/mL) |
Linear range | 0.1 – 8 µg/mL |
Recovery from the dilution water of OECD 210 test | 107% (0.3 mg/L) 96% (100 mg/L) |
Stability of the Stock Solution (5 ± 3 °C) | at least 2 days |
Stability in the Autosampler | at least 32 hours |
Stability in dilution water of OECD 210 test at 26 ± 1.5 °C | at least 7 days |
Table 3. Parameters of the calibration equations
Analytical occasion | Constant | X Coefficient | R. Squared |
November 12, 2021 | -264 | 93673 | 1.000 |
November 16, 2021 | 313 | 92970 | 1.000 |
November 19, 2021 | -171 | 94494 | 1.000 |
November 23, 2021 | -313 | 75429 | 1.000 |
November 26, 2021 | 197 | 74479 | 0.999 |
November 30, 2021 | -169 | 72770 | 1.000 |
December 03, 2021 | 197 | 73590 | 1.000 |
December 07, 2021 | 608 | 82128 | 0.999 |
December 13, 2021 | -449 | 82606 | 1.000 |
December 16, 2021 | 37 | 76878 | 1.000 |
Table 4: Concentrations measured at the renewal period of November 12-16, 2021
Nominal concentration mg/L | Start 1 | End 1 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.374 | 104 | 0.342 | 95 |
0.82 | 0.833 | 102 | 0.825 | 101 |
1.9 | 1.96 | 103 | 1.90 | 100 |
4.3 | 4.46 | 104 | 4.43 | 103 |
10 | 10.0 | 100 | 10.3 | 103 |
Table 5: Concentrations measured at the renewal period of November 16-19, 2021
Nominal concentration mg/L | Start 2 | End 2 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.360 | 100 | 0.387 | 108 |
0.82 | 0.849 | 103 | 0.886 | 108 |
1.9 | 1.97 | 104 | 2.06 | 108 |
4.3 | 4.60 | 107 | 4.69 | 109 |
10 | 10.8 | 108 | 11.0 | 110 |
Table 6: Concentrations measured at the renewal period of November 19-23, 2021
Nominal concentration mg/L | Start 3 | End 3 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.368 | 102 | 0.398 | 111 |
0.82 | 0.834 | 102 | 0.868 | 106 |
1.9 | 1.43 | 75 | 1.45 | 76 |
4.3 | 4.46 | 104 | 4.58 | 106 |
10 | 10.3 | 103 | 10.5 | 105 |
Table 7: Concentrations measured at the renewal period of November 23-26, 2021
Nominal concentration mg/L | Start 4 | End 4 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.359 | 100 | 0.377 | 105 |
0.82 | 0.894 | 109 | 0.961 | 117 |
1.9 | 1.86 | 98 | 2.00 | 105 |
4.3 | 4.23 | 98 | 4.57 | 106 |
10 | 9.73 | 97 | 10.9 | 109 |
Table 8: Concentrations measured at the renewal period of November 26-30, 2021
Nominal concentration mg/L | Start 5 | End 5 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.350 | 97 | 0.417 | 116 |
0.82 | 0.812 | 99 | 0.909 | 111 |
1.9 | 1.92 | 101 | 2.08 | 109 |
4.3 | 4.34 | 101 | 4.87 | 113 |
10 | 10.1 | 101 | 11.5 | 115 |
Table 9: Concentrations measured at the renewal period of November 30 – December 03, 2021
Nominal concentration mg/L | Start 6 | End 6 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.390 | 108 | 0.405 | 112 |
0.82 | 0.880 | 107 | 0.910 | 111 |
1.9 | 2.05 | 108 | 2.15 | 113 |
4.3 | 4.59 | 107 | 4.76 | 111 |
10 | 10.9 | 109 | 11.3 | 113 |
Table 10: Concentrations measured at the renewal period of December 03 - 07, 2021
Nominal concentration mg/L | Start 7 | End 7 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.374 | 104 | 0.351 | 97 |
0.82 | 0.843 | 103 | 0.803 | 98 |
1.9 | 1.97 | 104 | 1.92 | 101 |
4.3 | 4.50 | 105 | 4.38 | 102 |
10 | 10.4 | 104 | 10.2 | 102 |
Table 11: Concentrations measured at the renewal period of December 07 - 13, 2021
Nominal concentration mg/L | Start 8 | End 8 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.315 | 88 | 0.376 | 104 |
0.82 | 0.741 | 90 | 0.820 | 100 |
1.9 | 1.77 | 93 | 1.86 | 98 |
4.3 | 4.05 | 94 | 4.36 | 101 |
10 | 9.49 | 95 | 10.1 | 101 |
Table 12: Concentrations measured at the renewal period of December 13 - 16, 2021
Nominal concentration mg/L | Start 9 | End 9 | ||
Concentration, mean (mg/L) | % of the nominal | Concentration, mean (mg/L) | % of the nominal | |
0.36 | 0.317 | 88 | 0.368 | 102 |
0.82 | 0.702 | 86 | 0.825 | 101 |
1.9 | 1.63 | 86 | 1.92 | 101 |
4.3 | 3.71 | 86 | 4.36 | 101 |
10 | 8.80 | 88 | 10.8 | 108 |
Table 3. Preparation of the Stock and Test Solutions
Date | Relative Day | Test item weighed (g) | Final volume in the dilution water (mL) | Nominal concentration of the stock solution (mg/L) | Volume of the test solutions prepared from the stock solution (mL) |
2021.11.12. | 0 | 0.0102 | 100 | 100 | 500 |
2021.11.16. | 4 | 0.2007 | 200 | 1000 | 10000 |
2021.11.19. | 7 | 0.2011 | 200 | 1000 | 10000 |
2021.11.23. | 11 | 0.2001 | 200 | 1000 | 10000 |
2021.11.26. | 14 | 0.2001 | 200 | 1000 | 10000 |
2021.11.30. | 18 | 0.2043 | 200 | 1000 | 10000 |
2021.12.03. | 21 | 0.2000 | 200 | 1000 | 10000 |
2021.12.07. | 25 | 0.2003 | 200 | 1000 | 10000 |
2021.12.13. | 31 | 0.2007 | 200 | 1000 | 10000 |
Table 2. Components of the dilution (reconstituted, ISO) water
Substance | Nominal concentration in the reconstituted water (mg/L) | Nominal concentration in the dilution water (reconstituted water: purified water = 3:2) (mg/L) | |
CaCl2 x 2 H2O Batch No.: PP/2020/11623 Supplier: lach:ner Retest date: 31 March 2023 | 294 | 176.4 | |
MgSO4 x 7H2O Batch No.: PP/2020/01386 Supplier: lach:ner Retest date: 30 June 2024 | 123 | 73.8 | |
NaHCO3 Lot No.: 19I024103 Supplier: VWR Expiry date: 28 May 2024 | 65 | 39 | |
KCl Batch No.: K51320237 Supplier: Merck Expiry date: 30 April 2024 | 5.8 | 3.5 | |
Deionized water
| prepared with BWT Christ Ministil ion exchanger resin systems (Serial No.: 935455) in the laboratory of TOXI-COOP Zrt. |
Table 4 Temperature Ranges in the Test Chambers
Nominal concentrations (mg/L) | Parallels |
| Range (°C) |
|
Minimum | Maximum | Difference | ||
Control | 1 | 25.2 | 26.5 | 1.3 |
| 2 | 25.2 | 26.5 | 1.3 |
| 3 | 25.2 | 26.5 | 1.3 |
| 4 | 25.2 | 26.4 | 1.2 |
0.36 | 1 | 25.2 | 26.5 | 1.3 |
| 2 | 25.2 | 26.3 | 1.1 |
| 3 | 25.2 | 26.4 | 1.2 |
| 4 | 25.2 | 26.2 | 1.0 |
0.82 | 1 | 25.1 | 26.2 | 1.1 |
| 2 | 25.1 | 26.2 | 1.1 |
| 3 | 25.1 | 26.1 | 1.0 |
| 4 | 25.2 | 26.3 | 1.1 |
1.9 | 1 | 25.1 | 26.2 | 1.1 |
| 2 | 25.1 | 26.4 | 1.3 |
| 3 | 25.2 | 26.3 | 1.1 |
| 4 | 25.1 | 26.3 | 1.2 |
4.3 | 1 | 25.1 | 26.2 | 1.1 |
| 2 | 25.2 | 26.3 | 1.1 |
| 3 | 25.2 | 26.3 | 1.1 |
| 4 | 25.3 | 26.3 | 1.0 |
10 | 1 | 25.1 | 26.2 | 1.1 |
| 2 | 25.2 | 26.4 | 1.2 |
Table 5 Dissolved Oxygen Concentration Ranges in the Test Chambers
Nominal concentrations (mg/L) | Parallels |
| Range (% ASV) |
|
Minimum | Maximum | Difference | ||
Control | 1 | 62.9 | 93.4 | 30.5 |
| 2 | 63.4 | 92.5 | 29.1 |
| 3 | 63.2 | 93.4 | 30.2 |
| 4 | 68.0 | 93.2 | 25.2 |
0.36 | 1 | 65.8 | 92.1 | 26.3 |
| 2 | 63.9 | 92.1 | 28.2 |
| 3 | 66.7 | 94.3 | 27.6 |
| 4 | 62.4 | 93.6 | 31.2 |
0.82 | 1 | 61.4 | 93.1 | 31.7 |
| 2 | 60.8 | 93.7 | 32.9 |
| 3 | 61.2 | 93.6 | 32.4 |
| 4 | 60.7 | 93.4 | 32.7 |
1.9 | 1 | 63.4 | 92.0 | 28.6 |
| 2 | 62.8 | 93.2 | 30.4 |
| 3 | 65.7 | 92.3 | 26.6 |
| 4 | 64.4 | 93.1 | 28.7 |
4.3 | 1 | 61.7 | 94.3 | 32.6 |
| 2 | 61.3 | 93.8 | 32.5 |
| 3 | 60.9 | 92.0 | 31.1 |
| 4 | 62.4 | 92.1 | 29.7 |
10 | 1 | 62.1 | 93.7 | 31.6 |
| 2 | 63.0 | 95.4 | 32.4 |
| 3 | 64.6 | 94.2 | 29.6 |
| 4 | 62.7 | 94.1 | 31.4 |
ASV = Air Saturation Value
Table 6 pH Ranges in the Test Chambers
Nominal concentrations (mg/L) | Parallels |
| Range |
|
Minimum | Maximum | Difference | ||
Control | 1 | 7.06 | 7.81 | 0.75 |
| 2 | 7.09 | 7.81 | 0.72 |
| 3 | 7.10 | 7.90 | 0.80 |
| 4 | 7.12 | 7.90 | 0.78 |
0.36 | 1 | 7.14 | 7.90 | 0.76 |
| 2 | 7.08 | 7.90 | 0.82 |
| 3 | 7.11 | 7.90 | 0.79 |
| 4 | 7.08 | 7.90 | 0.82 |
0.82 | 1 | 7.09 | 7.84 | 0.75 |
| 2 | 7.06 | 7.83 | 0.77 |
| 3 | 7.07 | 7.81 | 0.74 |
| 4 | 7.11 | 7.81 | 0.70 |
1.9 | 1 | 7.04 | 7.73 | 0.69 |
| 2 | 7.07 | 7.73 | 0.66 |
| 3 | 7.08 | 7.74 | 0.66 |
| 4 | 7.10 | 7.73 | 0.63 |
4.3 | 1 | 7.03 | 7.83 | 0.80 |
| 2 | 7.10 | 7.83 | 0.73 |
| 3 | 7.12 | 7.83 | 0.71 |
| 4 | 7.09 | 7.83 | 0.74 |
10 | 1 | 7.06 | 7.82 | 0.76 |
| 2 | 7.05 | 7.82 | 0.77 |
| 3 | 7.08 | 7.82 | 0.74 |
| 4 | 7.12 | 7.82 | 0.70 |
Table 7 Total Hardness (mg/L CaCO3) of the Test Solutions
Nominal concentrations (mg/L) | Parallels |
| Days |
0a | 34b | ||
Control
| 1 2 | 169.1 | 169.1 169.1 |
| 3 |
| 178.0 |
| 4 |
| 178.0 |
0.36
| 1 2 | 160.2 | 178.0 178.0 |
| 3 |
| 178.0 |
| 4 |
| 178.0 |
0.82
| 1 2 | 169.1 | 178.0 178.0 |
| 3 |
| 178.0 |
| 4 |
| 178.0 |
1.9
| 1 2 | 160.2 | 178.0 169.1 |
| 3 |
| 178.0 |
| 4 |
| 178.0 |
4.3
| 1 2 | 160.2 | 178.0 178.0 |
| 3 |
| 178.0 |
| 4 |
| 178.0 |
10
| 1 2 | 160.2 | 178.0 178.0 |
| 3 |
| 178.0 |
| 4 |
| 178.0 |
a = measured in the test solutions used for treatment of the eggs b = measured in the test chambers at the end of the test
Table 8 Summarized Results of Analytical Determination of the Test Concentrations (Determined for 9 Renewal Periods)
Nominal Concentrations (mg/L) | Start of the Renewal Periods | End of the Renewal Periods | ||
Mean measured Concentrations (mg/L) with CI (95 %) | Percentage of the Nominal (%) | Mean Measured Concentrations (mg/L) with CI (95 %) | Percentage of the Nominal (%) | |
Control | not detected | - | not detected | - |
0.36 | 0.356 ± 0.020 | 99 | 0.380 ± 0.019 | 106 |
0.82 | 0.821 ± 0.048 | 100 | 0.867 ± 0.041 | 106 |
1.9 | 1.840 ± 0.153 | 97 | 1.927 ± 0.156 | 101 |
4.3 | 4.327 ± 0.223 | 101 | 4.556 ± 0.144 | 106 |
10 | 10.058 ± 0.505 | 101 | 10.733 ± 0.379 | 107 |
CI = Confidence Interval
Table 9 Evaluation of Data on Survival and Hatching
Nominal concentrations (mg/L) |
| Control |
|
| 0.36 |
|
| 0.82 |
|
| 1.9 |
|
| 4.3 |
|
| 10 |
| ||||||
Parallels | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 |
Number of eggs treated | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 |
Total no. of larvae hatched (Day 4) | 19 | 20 | 22 | 20 | 22 | 20 | 20 | 18 | 21 | 21 | 20 | 20 | 23 | 24 | 23 | 21 | 20 | 22 | 19 | 19 | 19 | 24 | 21 | 24 |
No. of fish at the end | 15 | 15 | 19 | 15 | 17 | 17 | 15 | 15 | 17 | 16 | 15 | 17 | 18 | 17 | 18 | 16 | 15 | 16 | 17 | 15 | 15 | 19 | 17 | 18 |
Cumulative mortality (Day 0-34) | 10 | 10 | 6 | 10 | 8 | 8 | 10 | 10 | 8 | 9 | 10 | 8 | 7 | 8 | 7 | 9 | 10 | 9 | 8 | 10 | 10 | 6 | 8 | 7 |
Sum: | 36 |
|
|
| 36 |
|
|
| 35 |
|
|
| 31 |
|
|
| 37 |
|
|
| 31 |
|
|
|
Mortality Day 0-4 Sum: | 6 19 | 5
| 3
| 5
| 3 | 5 | 5 | 7 | 4 18 | 4
| 5
| 5
| 2 | 1 | 2 | 4 | 5 20 | 3
| 6
| 6
| 6 | 1 | 4 | 1 |
20 |
|
|
| 9 |
|
|
| 12 |
|
|
| |||||||||||||
Mortality Day 5-34 | 4 | 5 | 3 | 5 | 5 | 3 | 5 | 3 | 4 | 5 | 5 | 3 | 5 | 7 | 5 | 5 | 5 | 6 | 2 | 4 | 4 | 5 | 4 | 6 |
Sum: | 17 |
|
|
| 16 |
|
|
| 17 |
|
|
| 22 |
|
|
| 17 |
|
|
| 19 |
|
|
|
Hatching success (%) | 76.0 | 80.0 | 88.0 | 80.0 | 88.0 | 80.0 | 80.0 | 72.0 | 84.0 | 84.0 | 80.0 | 80.0 | 92.0 | 96.0 | 92.0 | 84.0 | 80.0 | 88.0 | 76.0 | 76.0 | 76.0 | 96.0 | 84.0 | 96.0 |
Mean: | 81.0 |
|
|
| 80.0 |
|
|
| 82.0 |
|
|
| 91.0 |
|
|
| 80.0 |
|
|
| 88.0 |
|
|
|
Post-hatch success (%) | 78.9 | 75.0 | 86.4 | 75.0 | 77.3 | 85.0 | 75.0 | 83.3 | 81.0 | 76.2 | 75.0 | 85.0 | 78.3 | 70.8 | 78.3 | 76.2 | 75.0 | 72.7 | 89.5 | 78.9 | 78.9 | 79.2 | 81.0 | 75.0 |
Mean: | 78.8 |
|
|
| 80.2 |
|
|
| 79.3 |
|
|
| 75.9 |
|
|
| 79.0 |
|
|
| 78.5 |
|
|
|
Overall survival (%) | 60.0 | 60.0 | 76.0 | 60.0 | 68.0 | 68.0 | 60.0 | 60.0 | 68.0 | 64.0 | 60.0 | 68.0 | 72.0 | 68.0 | 72.0 | 64.0 | 60.0 | 64.0 | 68.0 | 60.0 | 60.0 | 76.0 | 68.0 | 72.0 |
Mean: | 64.0 |
|
|
| 64.0 |
|
|
| 65.0 |
|
|
| 69.0 |
|
|
| 63.0 |
|
|
| 69.0 |
|
|
|
Cumulative mortality (%) | 40.0 | 40.0 | 24.0 | 40.0 | 32.0 | 32.0 | 40.0 | 40.0 | 32.0 | 36.0 | 40.0 | 32.0 | 28.0 | 32.0 | 28.0 | 36.0 | 40.0 | 36.0 | 32.0 | 40.0 | 40.0 | 24.0 | 32.0 | 28.0 |
Mean: | 36.0 |
|
|
| 36.0 |
|
|
| 35.0 |
|
|
| 31.0 |
|
|
| 37.0 |
|
|
| 31.0 |
|
|
|
Table 10 Statistical Analysis of Data on Hatching
Nominal concentrations (mg/L) | Number of eggs treated | Day 3# Hatched |
Nonhatched | Day 4$ Hatched |
Nonhatched |
Control | 100 | 36 | 64 | 81 | 19 |
0.36 | 100 | 62 NS | 38 | 80 NS | 20 |
0.82 | 100 | 48NS | 52 | 82 NS | 18 |
1.9 | 100 | 44 NS | 56 | 91 NS | 9 |
4.3 | 100 | 39 NS | 61 | 80 NS | 20 |
10 | 100 | 40 NS | 60 | 88 NS | 12 |
Data were statistically evaluated by Chi2 2x2 Table Test with Bonferroni Correction (compared to the control). # = Start of hatching $ = End of hatching
NS = Not significant
Table 11 Statistical Analysis of Post-hatch Survival from Day 4 to Day 14
Nominal concentrations (mg/L) | Number of larvae survived (on Day 4) | Number of larvae survived (on Day 14) |
Control | 81 | 66 |
0.36 | 80 | 71 NS |
0.82 | 82 | 70 NS |
1.9 | 91 | 72 NS |
4.3 | 80 | 69 NS |
10 | 88 | 76 NS |
Data were statistically evaluated by Chi2 2x2 Table Test with Bonferroni Correction (compared to the control).
NS = not significant
Table 12 Statistical Analysis of Post-hatch Survival from Day 4 to Day 25
Nominal concentrations (mg/L) | Number of larvae survived (on Day 4) | Number of larvae survived (on Day 25) |
Control | 81 | 64 |
0.36 | 80 | 65 NS |
0.82 | 82 | 65 NS |
1.9 | 91 | 69 NS |
4.3 | 80 | 63 NS |
10 | 88 | 69 NS |
Data were statistically evaluated by Chi2 2x2 Table Test with Bonferroni Correction (compared to the control). NS = not significant
Table 13 Statistical Analysis of Post-hatch Survival from Day 4 to Day 34
Nominal concentrations (mg/L) | Number of larvae survived (on Day 4) | Number of larvae survived (on Day 34) |
Control | 81 | 64 |
0.36 | 80 | 64 NS |
0.82 | 82 | 65 NS |
1.9 | 91 | 69 NS |
4.3 | 80 | 63 NS |
10 | 88 | 69 NS |
Data were statistically evaluated by Chi2 2x2 Table Test with Bonferroni Correction (compared to the control). NS = not significant
Table 14 Statistical Analysis of the Body Weight Data
|
|
| Nominal concentrations (mg/L) |
| |||
| Parallels | Control | 0.36 | 0.82 | 1.9 | 4.3 | 10 |
1 2 3 4 | 0.0175 0.0237 0.0213 0.0254 | 0.0219 0.0194 0.0217 0.0256 | 0.0218 0.0206 0.0205 0.0222 | 0.0213 0.0226 0.0219 0.0231 | 0.0195 0.0207 0.0237 0.0227 | 0.0221 0.0260 0.0191 0.0206 | |
| Mean | 0.0220 | 0.0221 NS | 0.0213 NS | 0.0222 NS | 0.0216 NS | 0.0219 NS |
| SD | 0.003 | 0.003 | 0.001 | 0.001 | 0.002 | 0.003 |
| CV% | 15.6 | 11.6 | 4.0 | 3.5 | 8.8 | 13.5 |
Data were statistically evaluated by Dunett’s Multiple t-test (α = 0.050; one-sided smaller)
NS = not significant SD = Standard deviation CV = Coefficient of variation
Table 15 Statistical Analysis of the Body Length Data
|
|
| Nominal concentrations (mg/L) |
| |||
| Parallels | Control | 0.36 | 0.82 | 1.9 | 4.3 | 10 |
1 2 3 4 | 15,1 15.6 15.1 15.3 | 14.9 14.4 15.1 15.3 | 15.3 14.9 15.0 14.8 | 14.3 15.1 14.9 15.5 | 15.2 15.1 15.1 15.2 | 15.8 14.9 15.1 14.4 | |
| Mean | 15.28 | 14.93 NS | 15.00 NS | 14.95 NS | 15.15 NS | 15.05 NS |
| SD | 0.24 | 0.39 | 0.22 | 0.50 | 0.06 | 0.58 |
| CV% | 1.5 | 2.6 | 1.4 | 3.3 | 0.4 | 3.9 |
Data were statistically evaluated by Dunett’s Multiple t-test (α = 0.050; one-sided smaller)
NS = not significant SD = Standard deviation CV = Coefficient of variation
Table 16 Measured Temperature Values (°C)
Nominal concentrations [mg/L] | Parallels |
|
|
|
|
|
|
| Days/Measurements |
|
|
|
|
|
|
| |
0 T | 1 1 | 2 1 | 3 1 | 4 1 | 2 | 1 | 5 2 | 6 1 | 2 | 7 1 | 2 | 8 1 | 2 | 9 1 | 2 | ||
Control | 1 2 | 25.9 25.8 | 26.3 26.0 | 26.2 26.2 | 26.4 26.2 | 26.5 26.5 | 25.3 25.3 | 25.8 25.7 | 25.5 25.7 | 25.4 25.2 | 25.5 25.4 | 25.8 25.7 | 25.8 25.6 | 25.8 25.8 | 25.8 25.8 | 26.2 26.0 | 25.5 25.4 |
| 3 | 25.7 | 26.2 | 26.0 | 26.0 | 26.5 | 25.4 | 25.8 | 25.6 | 25.3 | 25.6 | 25.6 | 25.5 | 25.8 | 25.7 | 26.2 | 25.8 |
| 4 | 25.5 | 26.1 | 26.1 | 25.9 | 26.4 | 25.6 | 25.5 | 25.7 | 25.3 | 26.0 | 25.6 | 25.5 | 25.5 | 25.6 | 26.1 | 25.7 |
0.36 | 1 2 | 25.7 25.4 | 26.3 26.3 | 26.0 26.2 | 26.3 26.2 | 26.5 26.0 | 25.3 25.6 | 25.6 25.9 | 26.0 26.0 | 25.6 25.3 | 25.7 25.7 | 25.4 25.6 | 25.6 25.3 | 25.7 25.9 | 25.6 25.5 | 26.0 25.8 | 25.8 25.6 |
| 3 | 25.2 | 26.2 | 26.0 | 26.2 | 26.2 | 25.3 | 25.6 | 25.9 | 25.7 | 25.7 | 25.5 | 25.4 | 25.6 | 25.4 | 25.9 | 25.5 |
| 4 | 25.7 | 26.0 | 26.1 | 26.0 | 25.8 | 25.4 | 25.8 | 25.8 | 25.4 | 25.7 | 25.5 | 25.3 | 25.5 | 25.6 | 25.8 | 25.6 |
0.82 | 1 2 | 25.6 25.7 | 25.9 25.6 | 25.9 25.9 | 26.1 26.0 | 26.2 26.2 | 25.8 25.6 | 25.8 25.7 | 25.7 25.7 | 25.8 25.6 | 25.6 25.8 | 25.5 25.8 | 25.6 25.6 | 25.8 25.4 | 25.7 25.4 | 25.6 25.5 | 25.5 25.4 |
| 3 | 25.7 | 25.6 | 25.7 | 26.1 | 26.1 | 25.6 | 25.6 | 26.0 | 25.6 | 25.8 | 25.6 | 25.6 | 25.8 | 25.7 | 25.8 | 25.6 |
| 4 | 25.3 | 25.7 | 26.0 | 25.9 | 26.1 | 25.5 | 26.0 | 25.8 | 25.7 | 25.6 | 25.6 | 25.4 | 25.7 | 25.6 | 25.4 | 25.8 |
1.9 | 1 2 | 25.2 25.2 | 26.1 26.0 | 26.0 26.1 | 26.1 25.9 | 26.2 25.6 | 25.9 25.8 | 26.1 26.2 | 26.1 26.2 | 25.6 25.8 | 25.7 25.8 | 25.9 25.7 | 25.7 25.9 | 25.8 26.0 | 25.8 25.7 | 25.5 25.7 | 25.6 25.4 |
| 3 | 25.4 | 26.0 | 26.0 | 25.8 | 26.1 | 25.7 | 26.2 | 26.1 | 26.1 | 25.7 | 25.9 | 25.9 | 26.1 | 25.9 | 25.7 | 25.2 |
| 4 | 25.6 | 25.9 | 25.8 | 25.8 | 26.0 | 25.7 | 26.3 | 26.2 | 25.9 | 25.7 | 25.8 | 26.2 | 25.8 | 25.4 | 25.8 | 25.3 |
4.3 | 1 2 | 25.8 25.8 | 25.7 25.7 | 25.8 25.9 | 26.1 25.8 | 26.2 26.0 | 26.1 26.0 | 26.2 26.0 | 26.1 26.0 | 25.8 25.7 | 25.4 25.5 | 25.9 25.7 | 26.0 26.1 | 25.8 26.1 | 25.9 25.5 | 25.9 25.7 | 25.3 25.6 |
| 3 | 25.4 | 25.6 | 25.9 | 25.9 | 26.1 | 25.9 | 26.2 | 26.0 | 26.1 | 25.7 | 25.8 | 25.7 | 26.1 | 25.9 | 25.9 | 25.8 |
| 4 | 25.6 | 25.9 | 25.8 | 25.8 | 25.6 | 25.8 | 26.1 | 25.9 | 25.8 | 25.6 | 25.8 | 25.8 | 26.1 | 25.4 | 25.8 | 25.6 |
10 | 1 2 | 25.4 25.7 | 26.0 26.0 | 26.2 26.1 | 26.1 26.4 | 26.1 26.0 | 26.1 26.0 | 26.1 26.0 | 26.0 26.0 | 25.7 25.9 | 25.4 25.5 | 25.6 25.8 | 25.8 26.1 | 25.9 26.1 | 25.9 25.7 | 25.9 25.9 | 25.4 25.5 |
| 3 | 25.3 | 25.9 | 25.9 | 25.8 | 25.8 | 26.0 | 26.1 | 25.9 | 26.1 | 25.6 | 26.0 | 25.7 | 26.0 | 25.6 | 25.9 | 25.6 |
| 4 | 25.5 | 26.1 | 25.8 | 26.0 | 26.3 | 25.8 | 26.2 | 26.0 | 25.8 | 25.6 | 26.0 | 25.6 | 26.1 | 26.1 | 25.8 | 25.6 |
T = at the treatment
Nominal concentrations [mg/L] | Parallels |
|
|
|
|
|
|
| Days/Measurements |
|
|
|
|
|
|
| |
10 1 | 2 | 11 1 | 2 | 12 1 | 2 | 1 | 13 2 | 14 1 | 2 | 15 1 | 2 | 16 1 | 2 | 17 1 | 2 | ||
Control | 1 2 | 26.1 25.6 | 25.6 25.4 | 26.1 25.8 | 25.6 25.7 | 25.4 25.3 | 25.3 25.4 | 26.2 26.0 | 25.8 25.7 | 25.3 25.5 | 25.2 25.8 | 25.5 25.4 | 25.7 25.9 | 26.0 25.7 | 25.7 25.7 | 25.8 25.4 | 25.7 25.6 |
| 3 | 25.9 | 25.5 | 26.0 | 25.9 | 25.3 | 25.6 | 26.0 | 25.8 | 25.6 | 25.8 | 25.4 | 26.1 | 25.6 | 25.4 | 25.7 | 25.4 |
| 4 | 25.6 | 25.5 | 25.8 | 25.7 | 25.3 | 25.3 | 26.1 | 25.7 | 25.6 | 26.1 | 25.6 | 25.8 | 25.6 | 25.5 | 25.8 | 25.4 |
0.36 | 1 2 | 25.7 25.9 | 25.6 25.5 | 25.8 25.8 | 25.6 25.4 | 25.3 25.2 | 25.3 25.3 | 25.8 25.8 | 26.1 25.7 | 25.8 25.6 | 25.7 25.4 | 25.8 25.3 | 25.9 25.4 | 25.5 25.3 | 25.2 25.2 | 25.4 25.2 | 25.2 25.3 |
| 3 | 25.8 | 25.6 | 25.8 | 25.3 | 25.2 | 25.3 | 25.9 | 25.8 | 25.4 | 25.9 | 25.2 | 25.6 | 25.2 | 25.2 | 25.3 | 25.3 |
| 4 | 25.7 | 25.6 | 25.7 | 25.5 | 25.3 | 25.4 | 25.8 | 25.8 | 25.4 | 25.6 | 25.3 | 25.8 | 25.5 | 25.3 | 25.3 | 25.6 |
0.82 | 1 2 | 25.7 25.5 | 25.6 25.4 | 25.5 26.1 | 25.3 25.2 | 25.3 25.4 | 25.3 25.2 | 25.6 25.5 | 25.4 25.3 | 25.6 25.8 | 25.4 25.3 | 25.2 25.4 | 25.7 25.8 | 25.2 25.4 | 25.3 25.1 | 25.4 25.2 | 25.3 25.4 |
| 3 | 25.6 | 25.4 | 25.7 | 25.3 | 25.4 | 25.3 | 25.8 | 25.4 | 25.8 | 25.4 | 25.5 | 25.6 | 25.2 | 25.2 | 25.2 | 25.6 |
| 4 | 25.8 | 25.9 | 25.8 | 25.6 | 25.6 | 25.6 | 25.5 | 25.5 | 26.0 | 25.3 | 25.6 | 25.6 | 25.7 | 25.2 | 25.6 | 25.6 |
1.9 | 1 2 | 26.0 26.0 | 25.7 25.8 | 26.1 26.4 | 25.8 25.8 | 26.0 26.1 | 26.1 25.9 | 25.5 25.7 | 25.7 26.1 | 26.1 26.4 | 25.7 25.6 | 25.7 26.1 | 25.6 25.4 | 25.8 25.8 | 25.4 25.7 | 25.4 25.6 | 25.3 25.6 |
| 3 | 25.9 | 25.8 | 26.3 | 25.8 | 26.1 | 25.7 | 25.7 | 25.7 | 26.2 | 25.6 | 25.8 | 25.4 | 26.0 | 25.6 | 25.7 | 25.8 |
| 4 | 26.0 | 25.6 | 26.3 | 26.1 | 26.2 | 25.8 | 26.0 | 26.1 | 26.3 | 25.3 | 25.8 | 25.5 | 26.1 | 25.6 | 26.0 | 25.7 |
4.3 | 1 2 | 26.0 25.8 | 25.9 26.1 | 26.0 26.2 | 25.8 26.0 | 25.8 26.1 | 25.6 25.7 | 25.9 25.7 | 25.2 25.4 | 26.1 26.2 | 25.3 25.4 | 26.2 26.3 | 25.8 25.7 | 26.2 26.2 | 25.6 25.8 | 26.0 26.1 | 25.3 25.7 |
| 3 | 26.1 | 26.0 | 26.2 | 25.8 | 26.2 | 25.7 | 25.9 | 25.8 | 26.3 | 25.3 | 26.1 | 25.4 | 26.3 | 25.7 | 25.6 | 25.6 |
| 4 | 25.9 | 25.8 | 26.2 | 26.2 | 26.1 | 26.0 | 25.8 | 26.1 | 26.3 | 25.8 | 26.1 | 25.4 | 26.2 | 25.8 | 26.1 | 25.8 |
10 | 1 2 | 25.7 25.8 | 25.5 25.5 | 26.0 26.1 | 25.8 26.0 | 25.8 26.1 | 25.6 26.1 | 25.9 25.9 | 25.4 25.9 | 26.1 26.2 | 25.8 25.7 | 26.1 26.3 | 25.3 25.7 | 25.9 25.9 | 25.9 25.9 | 26.0 26.1 | 25.3 25.7 |
| 3 | 25.8 | 25.6 | 26.2 | 25.8 | 26.3 | 26.0 | 25.9 | 25.8 | 26.3 | 25.7 | 26.3 | 25.7 | 25.9 | 25.7 | 25.6 | 25.6 |
| 4 | 26.1 | 25.6 | 26.3 | 26.2 | 26.1 | 26.0 | 26.0 | 26.1 | 26.4 | 25.4 | 26.3 | 25.7 | 26.2 | 25.8 | 26.1 | 25.8 |
Nominal concentrations [mg/L] | Parallels |
|
|
|
|
|
|
| Days/Measurements |
|
|
|
|
|
|
| |
18 1 | 2 | 19 1 | 2 | 20 1 | 2 | 1 | 21 2 | 22 1 | 2 | 23 1 | 2 | 24 1 | 2 | 25 1 | 2 | ||
Control | 1 2 | 26.0 26.0 | 25.5 25.5 | 25.4 25.3 | 26.0 25.3 | 25.7 25.3 | 25.6 25.8 | 26.1 25.7 | 25.7 26.0 | 26.3 26.1 | 25.2 25.2 | 26.2 25.9 | 25.7 26.0 | 26.1 25.7 | 25.9 25.9 | 26.1 25.4 | 25.4 25.2 |
| 3 | 26.1 | 25.5 | 25.4 | 25.8 | 25.3 | 25.4 | 25.6 | 25.9 | 26.3 | 25.2 | 26.1 | 25.3 | 26.1 | 25.7 | 25.4 | 26.0 |
| 4 | 25.9 | 25.3 | 25.2 | 25.7 | 25.8 | 25.8 | 26.1 | 26.2 | 25.4 | 25.3 | 26.4 | 25.8 | 26.2 | 25.9 | 25.8 | 25.3 |
0.36 | 1 2 | 25.3 25.2 | 25.9 25.3 | 25.5 25.4 | 25.8 25.9 | 25.7 26.0 | 25.2 25.2 | 25.9 26.2 | 26.1 26.1 | 25.4 25.8 | 25.2 25.2 | 26.4 26.3 | 25.7 25.8 | 26.2 26.0 | 26.0 26.0 | 25.6 25.8 | 25.8 25.7 |
| 3 | 25.3 | 25.5 | 25.4 | 25.4 | 25.3 | 25.2 | 25.7 | 25.7 | 25.6 | 25.3 | 26.4 | 25.9 | 26.2 | 26.1 | 25.4 | 25.8 |
| 4 | 25.2 | 25.6 | 25.6 | 25.4 | 25.8 | 25.4 | 26.1 | 25.9 | 25.8 | 25.2 | 26.2 | 25.4 | 26.0 | 25.9 | 25.8 | 25.9 |
0.82 | 1 2 | 25.4 25.4 | 25.2 25.3 | 25.3 25.5 | 25.8 25.6 | 25.7 25.8 | 25.3 25.2 | 25.4 25.4 | 25.3 26.0 | 25.4 25.8 | 25.3 25.4 | 25.8 26.1 | 26.2 26.1 | 25.3 25.8 | 25.7 25.9 | 25.6 25.8 | 25.4 25.5 |
| 3 | 25.6 | 25.4 | 25.6 | 25.8 | 25.9 | 25.1 | 25.8 | 25.3 | 25.6 | 25.2 | 26.0 | 25.9 | 25.7 | 25.9 | 25.9 | 25.6 |
| 4 | 25.6 | 25.8 | 26.0 | 25.4 | 25.4 | 25.2 | 25.6 | 25.8 | 25.8 | 25.3 | 25.8 | 26.3 | 25.8 | 26.0 | 26.3 | 26.0 |
1.9 | 1 2 | 25.3 25.2 | 25.6 25.9 | 26.1 26.1 | 25.8 25.6 | 25.1 25.1 | 25.1 25.2 | 25.8 25.4 | 25.7 25.8 | 25.5 25.7 | 25.3 25.2 | 25.4 25.6 | 26.2 26.1 | 25.9 25.4 | 25.8 25.4 | 26.2 26.1 | 25.4 25.6 |
| 3 | 25.3 | 25.7 | 26.0 | 25.8 | 25.2 | 25.3 | 25.8 | 25.9 | 25.6 | 25.3 | 25.6 | 25.8 | 25.6 | 25.4 | 25.7 | 25.6 |
| 4 | 25.2 | 25.9 | 26.2 | 25.1 | 25.3 | 25.4 | 25.6 | 26.3 | 25.7 | 25.2 | 25.7 | 26.0 | 25.8 | 25.8 | 25.8 | 25.7 |
4.3 | 1 2 | 26.1 26.0 | 26.0 26.1 | 26.2 26.1 | 25.1 25.2 | 25.1 25.6 | 25.4 25.8 | 25.8 25.8 | 26.2 26.1 | 25.8 26.0 | 26.0 25.3 | 25.7 26.1 | 25.4 25.4 | 25.7 26.1 | 25.6 25.8 | 25.9 26.3 | 25.7 26.1 |
| 3 | 25.9 | 25.8 | 26.2 | 25.2 | 25.5 | 25.6 | 25.7 | 25.9 | 26.1 | 25.8 | 26.1 | 25.6 | 26.0 | 25.4 | 26.2 | 25.8 |
| 4 | 25.8 | 26.0 | 25.9 | 25.3 | 25.3 | 25.8 | 25.8 | 26.3 | 26.1 | 25.7 | 26.1 | 25.7 | 26.1 | 25.8 | 26.1 | 25.7 |
10 | 1 2 | 25.3 25.4 | 26.0 26.1 | 26.2 26.1 | 25.1 25.2 | 25.1 25.6 | 25.4 25.8 | 25.6 25.8 | 26.2 26.1 | 25.8 26.0 | 25.8 25.9 | 25.7 26.1 | 25.4 25.4 | 25.7 26.1 | 25.6 25.8 | 25.9 26.3 | 25.8 25.6 |
| 3 | 25.2 | 25.8 | 26.0 | 25.2 | 25.5 | 25.6 | 25.7 | 25.5 | 26.1 | 25.4 | 26.1 | 25.6 | 26.0 | 25.8 | 26.2 | 25.8 |
| 4 | 25.5 | 26.0 | 26.2 | 25.3 | 25.3 | 25.8 | 25.8 | 25.9 | 26.1 | 25.5 | 26.1 | 25.7 | 26.1 | 26.0 | 26.1 | 26.2 |
Nominal concentrations [mg/L] | Parallels |
|
|
|
| Days/Measurements |
|
|
| ||
26 1 | 27 1 | 28 1 | 29 1 | 30 1 | 31 1 | 2 | 32 1 | 33 1 | 34 1 | ||
Control | 1 2 | 25.6 25.6 | 25.7 25.3 | 25.2 25.3 | 25.7 25.4 | 25.6 25.4 | 25.6 25.3 | 25.6 25.4 | 26.0 25.8 | 26.2 26.0 | 26.0 25.9 |
| 3 | 25.7 | 25.6 | 25.6 | 25.6 | 25.6 | 25.6 | 25.7 | 25.8 | 26.1 | 26.0 |
| 4 | 25.3 | 25.8 | 25.3 | 25.8 | 25.2 | 25.9 | 25.7 | 25.9 | 26.3 | 26.1 |
0.36 | 1 2 | 25.5 25.2 | 25.8 26.1 | 25.6 25.2 | 25.9 26.0 | 25.3 25.3 | 25.9 25.8 | 25.6 25.6 | 26.1 26.2 | 25.6 25.2 | 26.2 26.2 |
| 3 | 25.3 | 25.7 | 25.2 | 25.2 | 25.3 | 25.6 | 25.2 | 25.8 | 25.6 | 26.0 |
| 4 | 25.6 | 25.8 | 26.1 | 25.3 | 25.4 | 25.7 | 25.3 | 26.0 | 25.3 | 26.1 |
0.82 | 1 2 | 25.3 25.6 | 25.6 25.7 | 26.2 25.8 | 25.6 25.3 | 25.5 25.8 | 25.2 25.2 | 25.1 25.6 | 25.4 25.7 | 26.0 25.9 | 25.7 25.8 |
| 3 | 25.7 | 25.3 | 26.0 | 25.6 | 25.8 | 25.4 | 25.4 | 25.4 | 25.6 | 25.6 |
| 4 | 25.4 | 25.5 | 25.4 | 25.7 | 25.6 | 25.5 | 25.7 | 25.4 | 25.7 | 25.7 |
1.9 | 1 2 | 25.8 26.0 | 25.2 25.3 | 25.7 25.4 | 25.6 25.3 | 25.6 26.1 | 25.7 25.3 | 25.8 25.9 | 25.4 25.3 | 25.5 25.5 | 25.3 25.4 |
| 3 | 25.3 | 25.6 | 25.2 | 26.1 | 26.1 | 25.8 | 25.7 | 25.3 | 25.7 | 25.6 |
| 4 | 25.3 | 25.3 | 25.2 | 26.2 | 25.6 | 25.8 | 26.0 | 25.3 | 25.4 | 25.4 |
4.3 | 1 2 | 25.4 25.8 | 25.6 25.7 | 25.2 25.3 | 25.8 26.0 | 25.3 25.6 | 25.6 25.2 | 25.1 25.7 | 25.4 25.5 | 25.7 25.6 | 25.6 25.6 |
| 3 | 25.8 | 25.6 | 25.6 | 25.4 | 25.3 | 25.6 | 25.5 | 25.8 | 26.2 | 26.1 |
| 4 | 25.9 | 25.9 | 25.7 | 25.7 | 25.6 | 25.3 | 25.7 | 25.8 | 25.9 | 26.1 |
10 | 1 2 | 25.7 25.7 | 25.6 25.7 | 25.2 25.3 | 25.4 25.3 | 25.7 25.6 | 26.0 26.1 | 25.4 25.2 | 25.4 25.5 | 25.7 25.6 | 25.6 25.6 |
| 3 | 25.5 | 25.6 | 25.6 | 25.2 | 25.7 | 25.9 | 25.3 | 25.7 | 26.2 | 26.1 |
| 4 | 25.3 | 25.9 | 25.7 | 25.4 | 25.6 | 25.8 | 25.2 | 25.6 | 25.9 | 26.1 |
1 = before renewal of the test solution
Table 17 Measured Minimum-Maximum Temperatures (°C) in the Concurrent Test Chamber
|
|
|
|
|
|
|
| Days |
|
|
|
|
|
|
|
0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |
Minimum | - | 25.3 | 25.4 | 25.2 | 25.2 | 25.2 | 25.1 | 25.2 | 25.2 | 25.1 | 25.3 | 25.2 | 25.1 | 25.2 | 25.1 |
Maximum | - | 26.4 | 26.4 | 26.4 | 26.2 | 26.5 | 26.5 | 26.5 | 26.4 | 26.4 | 26.4 | 26.4 | 26.2 | 26.2 | 26.5 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| Days |
|
|
|
|
|
|
|
15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 | 27 | 28 | 29 | |
Minimum | 25.3 | 25.2 | 25.1 | 25.2 | 25.1 | 25.3 | 25.1 | 25.2 | 25.4 | 25.5 | 25.2 | 25.1 | 25.2 | 25.1 | 25.1 |
Maximum | 26.5 | 26.3 | 26.3 | 26.1 | 26.2 | 26.4 | 26.3 | 26.5 | 26.4 | 26.4 | 26.3 | 26.5 | 26.4 | 26.4 | 26.3 |
|
|
| Days |
|
|
30 | 31 | 32 | 33 | 34 | |
Minimum | 25.1 | 25.2 | 25.1 | 25.2 | 25.3 |
Maximum | 26.4 | 26.0 | 26.3 | 26.0 | 26.3 |
Table 18 Measured Dissolved Oxygen Concentrations (% ASV)
3 90.2 89.5 92.0 63.2 89.9 71.4 | 89.8 | 73.5 | 91.5 | 73.0 | 93.4 | 71.8 | 92.5 | 72.1 90.5 72.1 93.0 75.4 73.0 66.7 70.0 | 71.0 |
4 73.2 | 92.1 | 70.5 | 92.0 | 70.9 | 93.0 | 75.0 | 92.9 |
|
|
| 75.0 | 89.5 | 73.3 | 91.0 | 69.8 | 92.1 | 74.2 | 91.4 |
91.4 | 74.7 | 90.4 | 76.2 | 90.7 | 71.0 | 91.1 | 74.0 | 92.1 |
92.4 | 72.6 | 92.9 | 69.9 | 89.9 | 74.6 | 89.7 | 74.6 | 94.3 |
| 71.4 | 92.0 | 73.2 | 90.0 | 72.0 | 92.1 | 71.2 | 90.8 |
| 70.9 | 89.5 | 69.7 | 90.5 | 72.5 | 93.0 | 71.0 | 90.6 |
92.7 | 72.1 | 89.9 | 72.0 | 92.0 | 72.8 | 92.5 | 71.9 | 92.7 |
90.9 | 73.0 | 91.8 | 72.4 | 92.1 | 70.1 | 90.9 | 72.8 | 93.6 |
| 74.1 | 92.0 | 73.4 | 90.0 | 73.2 | 90.4 | 75.0 | 93.4 |
| 70.6 | 89.2 | 70.4 | 89.9 | 69.9 | 90.4 | 70.4 | 90.7 |
93.2 | 71.8 | 90.3 | 69.9 | 91.9 | 70.1 | 91.6 | 74.3 | 90.2 |
91.7 | 73.8 | 91.2 | 71.0 | 90.8 | 72.3 | 92.3 | 73.9 | 90.5 |
| 73.1 | 91.3 | 73.5 | 89.8 | 69.8 | 93.1 | 73.1 | 92.3 |
| 75.4 | 92.1 | 72.1 | 90.2 | 71.0 | 94.3 | 76.0 | 90.9 |
93.8 | 70.9 | 92.0 | 71.1 | 91.6 | 72.1 | 93.8 | 74.1 | 91.1 |
91.6 | 71.3 | 91.6 | 73.3 | 91.2 | 70.8 | 90.1 | 73.3 | 91.3 |
| 72.6 | 90.8 | 69.8 | 90.0 | 69.9 | 90.1 | 72.4 | 90.8 |
0.36 | 1 2 3 4 |
0.82 | 1 2 3 4 |
1.9 | 1 2 3 4 |
4.3 | 1 2 3 4 |
91.3 89.7 90.3 63.9 71.2 89.9 75.1 92.0 77.3 71.9 68.1 70.2 69.2
91.3 89.7 92.9 66.7 70.0 90.4 74.2 90.8 76.2 72.2 67.0 70.1 70.4
89.9 88.9 91.8 60.8 69.9 93.7 71.8 91.3 75.8 70.2 69.7 70.3 70.9
89.9 88.9 92.0 61.2 72.5 91.1 73.0 92.0 77.0 71.0 67.8 71.0 72.4
92.0 88.4 89.9 62.8 71.6 93.1 72.5 90.5 75.8 71.1 71.3 70.2 70.1
92.0 88.4 91.0 65.7 72.2 90.2 72.2 90.0 74.3 69.9 70.0 70.3 69.8
91.1 87.2 92.3 61.3 72.5 90.1 75.8 91.6 73.6 72.0 71.2 68.7 69.6
91.1 87.2 91.4 60.9 71.4 92.0 73.0 89.9 74.9 70.5 70.0 70.2 70.4
10
Table 19 Measured pH values
Nominal concentrations [mg/L] | Parallels |
|
|
|
|
|
| Days |
|
|
|
|
|
0 * | 4 a* | 4 a | 5 b | 11 a | 12 b | 18 a | 19 b | 25 a | 26
| 29
| 34
| ||
Control | 1 2 | 7.81 7.81 | 7.79 7.79 | 7.76 7.81 | 7.20 7.18 | 7.74 7.80 | 7.35 7.30 | 7.65 7.79 | 7.18 7.11 | 7.72 7.81 | 7.06 7.20 | 7.21 7.09 | 7.25 7.18 |
| 3 | 7.81 | 7.79 | 7.82 | 7.19 | 7.90 | 7.41 | 7.89 | 7.10 | 7.76 | 7.20 | 7.19 | 7.13 |
| 4 | 7.81 | 7.79 | 7.88 | 7.22 | 7.82 | 7.38 | 7.90 | 7.23 | 7.85 | 7.22 | 7.13 | 7.12 |
0.36 | 1 2 | 7.90 7.90 | 7.83 7.83 | 7.85 7.76 | 7.19 7.30 | 7.89 7.85 | 7.38 7.32 | 7.90 7.84 | 7.16 7.15 | 7.85 7.80 | 7.39 7.08 | 7.15 7.13 | 7.14 7.11 |
| 3 | 7.90 | 7.83 | 7.77 | 7.29 | 7.76 | 7.33 | 7.85 | 7.15 | 7.79 | 7.24 | 7.15 | 7.11 |
| 4 | 7.90 | 7.83 | 7.73 | 7.25 | 7.81 | 7.25 | 7.90 | 7.24 | 7.76 | 7.24 | 7.08 | 7.13 |
0.82 | 1 2 3 | 7.81 7.81 7.81 | 7.81 7.81 7.81 | 7.74 7.71 7.67 | 7.20 7.44 7.35 | 7.78 7.72 7.70 | 7.27 7.26 7.31 | 7.78 7.83 7.75 | 7.21 7.19 7.10 | 7.84 7.81 7.76 | 7.28 7.16 7.07 | 7.19 7.06 7.11 | 7.09 7.11 7.20 |
| 4 | 7.81 | 7.81 | 7.60 | 7.41 | 7.69 | 7.38 | 7.76 | 7.11 | 7.69 | 7.17 | 7.12 | 7.16 |
1.9 | 1 2 3 | 7.70 7.70 7.70 | 7.73 7.73 7.73 | 7.69 7.68 7.74 | 7.23 7.41 7.25 | 7.68 7.72 7.70 | 7.22 7.22 7.21 | 7.73 7.68 7.71 | 7.10 7.10 7.08 | 7.65 7.65 7.71 | 7.04 7.24 7.16 | 7.10 7.12 7.13 | 7.09 7.07 7.18 |
| 4 | 7.70 | 7.73 | 7.71 | 7.21 | 7.73 | 7.29 | 7.71 | 7.15 | 7.70 | 7.12 | 7.20 | 7.10 |
4.3 | 1 2 3 | 7.83 7.83 7.83 | 7.76 7.76 7.76 | 7.72 7.71 7.71 | 7.32 7.33 7.25 | 7.69 7.72 7.73 | 7.19 7.23 7.31 | 7.75 7.71 7.71 | 7.09 7.15 7.12 | 7.73 7.83 7.74 | 7.28 7.31 7.36 | 7.03 7.19 7.12 | 7.12 7.10 7.13 |
| 4 | 7.83 | 7.76 | 7.68 | 7.29 | 7.73 | 7.32 | 7.76 | 7.12 | 7.69 | 7.09 | 7.12 | 7.09 |
10 | 1 2 3 | 7.82 7.82 7.82 | 7.68 7.68 7.68 | 7.70 7.65 7.69 | 7.22 7.23 7.23 | 7.68 7.70 7.75 | 7.23 7.25 7.28 | 7.72 7.69 7.72 | 7.18 7.05 7.18 | 7.71 7.66 7.66 | 7.10 7.21 7.25 | 7.06 7.19 7.08 | 7.09 7.08 7.09 |
| 4 | 7.82 | 7.68 | 7.62 | 7.40 | 7.69 | 7.32 | 7.72 | 7.12 | 7.68 | 7.21 | 7.16 | 7.12 |
Table 20 Number of Larvae Hatched and Duration of Hatching
Nominal concentrations [mg/L] | Parallels | Number of eggs treated |
|
| Days |
|
|
0 | 1 | 2 | 3 | 4 | |||
Control | 1 2 | 25 25 | 0 0 | 0 0 | 0 0 | 10 8 | 19 20 |
| 3 | 25 | 0 | 0 | 0 | 12 | 22 |
| 4 | 25 | 0 | 0 | 0 | 6 | 20 |
0.36 | 1 2 | 25 25 | 0 0 | 0 0 | 0 0 | 19 18 | 22 20 |
| 3 | 25 | 0 | 0 | 0 | 16 | 20 |
| 4 | 25 | 0 | 0 | 0 | 9 | 18 |
0.82 | 1 2 | 25 25 | 0 0 | 0 0 | 0 0 | 11 14 | 21 21 |
| 3 | 25 | 0 | 0 | 0 | 7 | 20 |
| 4 | 25 | 0 | 0 | 0 | 16 | 20 |
1.9 | 1 2 | 25 25 | 0 0 | 0 0 | 0 0 | 8 11 | 23 24 |
| 3 | 25 | 0 | 0 | 0 | 18 | 23 |
| 4 | 25 | 0 | 0 | 0 | 7 | 21 |
4.3 | 1 2 | 25 25 | 0 0 | 0 0 | 0 0 | 3 16 | 20 22 |
| 3 | 25 | 0 | 0 | 0 | 8 | 19 |
| 4 | 25 | 0 | 0 | 0 | 12 | 19 |
10 | 1 2 | 25 25 | 0 0 | 0 0 | 0 0 | 12 4 | 19 24 |
| 3 | 25 | 0 | 0 | 0 | 11 | 21 |
| 4 | 25 | 0 | 0 | 0 | 13 | 24 |
Table 21 Number of Living and Dead Embryo/Larvae/Fish and Number of Abnormalities Observed During the Test
Nominal Concentrations (mg/L) |
|
|
| Control |
|
|
|
|
|
| 0.36 |
|
| |||||||
Parallels |
| 1 |
| 2 |
|
| 3 |
|
| 4 |
| 1 |
| 2 |
| 3 |
|
| 4 | |
Days | S |
| M | S | M | S |
| M | S |
| M | S | M | S | M | S | M | S | M | |
0 | 25 |
| 0 | 25 | 0 | 25 |
| 0 | 25 |
| 0 | 25 |
| 0 | 25 | 0 | 25 | 0 | 25 | 0 |
1 | 19 |
| 6 | 20 | 5 | 22 |
| 3 | 20 |
| 5 | 22 |
| 3 | 20 | 5 | 20 | 5 | 19 | 6 |
2 | 19 |
| 0 | 20 | 0 | 22 |
| 0 | 20 |
| 0 | 22 |
| 0 | 20 | 0 | 20 | 0 | 18 | 1 |
3 | 19 |
| 0 | 20 | 0 | 22 |
| 0 | 20 |
| 0 | 22 |
| 0 | 20 | 0 | 20 | 0 | 18 | 0 |
4 | 19 |
| 0 | 20 | 0 | 22 |
| 0 | 20 |
| 0 | 22 |
| 0 | 20 | 0 | 20 (2) | 0 | 18 | 0 |
5 | 19 |
| 0 | 20 | 0 | 22 |
| 0 | 20 |
| 0 | 22 |
| 0 | 20 | 0 | 20 (2) | 0 | 18 | 0 |
6 | 19 |
| 0 | 19 | 1 | 22 |
| 0 | 20 |
| 0 | 21 | 1 | 20 | 0 | 20 (2) | 0 | 18 | 0 | |
7 | 18 (1) |
| 1 | 19 | 0 | 21 |
| 1 | 18 |
| 2 | 21 |
| 0 | 20 | 0 | 20 (2) | 0 | 17 | 1 |
8 | 18 (1) |
| 0 | 18 | 1 | 21 |
| 0 | 18 |
| 0 | 21 | 0 | 20 | 0 | 20 (2) | 0 | 17 | 0 | |
9 | 18 (1) |
| 0 | 18 | 0 | 21 |
| 0 | 18 |
| 0 | 20 |
| 1 | 20 | 0 | 20 (2) | 0 | 16 | 1 |
10 | 18 (1) |
| 0 | 18 | 0 | 21 |
| 0 | 18 |
| 0 | 20 | 0 | 20 | 0 | 18 | 2 | 16 | 0 | |
11 | 17 |
| 1 | 17 | 1 | 20 |
| 1 | 17 |
| 1 | 20 |
| 0 | 20 | 0 | 18 | 0 | 16 | 0 |
12 | 17 |
| 0 | 17 | 0 | 20 |
| 0 | 16 |
| 1 | 20 | 0 | 20 | 0 | 18 | 0 | 16 | 0 | |
13 | 16 |
| 1 | 17 | 0 | 20 |
| 0 | 16 |
| 0 | 20 |
| 0 | 20 | 0 | 18 | 0 | 16 | 0 |
14 | 16 |
| 0 | 16 | 1 | 19 |
| 1 | 15 |
| 1 | 19 | 1 | 19 | 1 | 17 | 1 | 16 | 0 | |
15 | 16 |
| 0 | 16 | 0 | 19 |
| 0 | 15 |
| 0 | 19 (1) |
| 0 | 19 (1) | 0 | 17 (1) | 0 | 16 | 0 |
16 | 16 |
| 0 | 16 (1) | 0 | 19 |
| 0 | 15 |
| 0 | 19 (1) | 0 | 19 (1) | 0 | 17 (1) | 0 | 16 | 0 | |
17 | 16 |
| 0 | 16 (1) | 0 | 19 |
| 0 | 15 |
| 0 | 19 (1) |
| 0 | 19 (1) | 0 | 17 (1) | 0 | 16 | 0 |
18 | 15 |
| 1 | 16 (1) | 0 | 19 |
| 0 | 15 |
| 0 | 19 (1) | 0 | 19 (1) | 0 | 17 (1) | 0 | 16 | 0 | |
19 | 15 |
| 0 | 16 (1) | 0 | 19 |
| 0 | 15 |
| 0 | 19 (1) |
| 0 | 19 (1) | 0 | 17 (1) | 0 | 16 | 0 |
20 | 15 |
| 0 | 16 (1) | 0 | 19 |
| 0 | 15 |
| 0 | 18 | 1 | 18 | 1 | 16 | 1 | 16 | 0 | |
21 | 15 |
| 0 | 16 (1) | 0 | 19 |
| 0 | 15 |
| 0 | 18 |
| 0 | 18 | 0 | 16 | 0 | 16 | 0 |
22 | 15 |
| 0 | 15 | 1 | 19 |
| 0 | 15 |
| 0 | 18 | 0 | 18 | 0 | 15 | 1 | 16 | 0 | |
23 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 18 |
| 0 | 18 | 0 | 15 | 0 | 15 | 1 |
24 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 | 1 | 18 | 0 | 15 | 0 | 15 | 0 | |
25 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 |
| 0 | 18 | 0 | 15 | 0 | 15 | 0 |
26 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 15 | 0 | 15 | 0 | |
27 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 |
| 0 | 18 | 0 | 15 | 0 | 15 | 0 |
28 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 15 | 0 | 15 | 0 | |
29 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 |
| 0 | 18 | 0 | 15 | 0 | 15 | 0 |
30 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 | 0 | 17 | 1 | 15 | 0 | 15 | 0 | |
31 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 |
| 0 | 17 | 0 | 15 | 0 | 15 | 0 |
32 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 | 0 | 17 | 0 | 15 | 0 | 15 | 0 | |
33 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 |
| 0 | 17 | 0 | 15 | 0 | 15 | 0 |
34 | 15 |
| 0 | 15 | 0 | 19 |
| 0 | 15 |
| 0 | 17 | 0 | 17 | 0 | 15 | 0 | 15 | 0 |
Table 21 (continued) Number of Living and Dead Embryo/Larvae/Fish and Number of Abnormalities Observed During the Test
Nominal Concentrations (mg/L) |
|
|
| 0.82 |
|
|
|
|
| 1.9 |
|
| |||||||
Parallels |
| 1 |
| 2 |
|
| 3 |
| 4 |
| 1 |
| 2 |
| 3 |
|
| 4 | |
Days | S |
| M | S | M | S |
| M | S | M | S | M | S | M | S | M | S | M | |
0 | 25 |
| 0 | 25 | 0 | 25 |
| 0 | 25 | 0 | 25 |
| 0 | 25 | 0 | 25 | 0 | 25 | 0 |
1 | 23 |
| 2 | 21 | 4 | 20 |
| 5 | 20 | 5 | 23 |
| 2 | 24 | 1 | 24 | 1 | 23 | 2 |
2 | 23 (1) |
| 0 | 21 (1) | 0 | 20 |
| 0 | 20 | 0 | 23 |
| 0 | 24 (1) | 0 | 24 | 0 | 23 (1) | 0 |
3 | 22 |
| 1 | 21 (1) | 0 | 20 |
| 0 | 20 (1) | 0 | 23 |
| 0 | 24 (1) | 0 | 24 | 0 | 21 | 2 |
4 | 21 (2) |
| 1 | 21 (1) | 0 | 20 (1) |
| 0 | 20 (1) | 0 | 23 (3) |
| 0 | 24 (3) | 0 | 23 (2) | 1 | 21 (2) | 0 |
5 | 21 (2) |
| 0 | 21 (1) | 0 | 20 (1) |
| 0 | 20 (1) | 0 | 22 (2) |
| 1 | 23 (2) | 1 | 22 (1) | 1 | 20 (1) | 1 |
6 | 21 (2) |
| 0 | 21 (1) | 0 | 19 |
| 1 | 20 (1) | 0 | 21 (2) | 1 | 21 | 2 | 21 | 1 | 20 (1) | 0 | |
7 | 20 (1) |
| 1 | 20 | 1 | 19 |
| 0 | 19 | 1 | 19 |
| 2 | 20 | 1 | 21 | 0 | 18 | 2 |
8 | 20 |
| 0 | 20 | 0 | 19 |
| 0 | 19 | 0 | 19 | 0 | 20 | 0 | 20 | 1 | 18 | 0 | |
9 | 18 |
| 2 | 19 | 1 | 19 |
| 0 | 18 | 1 | 19 |
| 0 | 20 | 0 | 20 | 0 | 18 | 0 |
10 | 18 |
| 0 | 18 | 1 | 17 |
| 2 | 18 | 0 | 19 | 0 | 19 | 1 | 20 | 0 | 18 | 0 | |
11 | 18 |
| 0 | 18 | 0 | 17 |
| 0 | 18 | 0 | 19 |
| 0 | 19 | 0 | 20 | 0 | 18 | 0 |
12 | 18 |
| 0 | 18 | 0 | 17 |
| 0 | 18 | 0 | 19 | 0 | 19 | 0 | 20 | 0 | 18 | 0 | |
13 | 18 |
| 0 | 18 | 0 | 17 |
| 0 | 18 | 0 | 19 |
| 0 | 19 | 0 | 20 | 0 | 18 | 0 |
14 | 18 |
| 0 | 18 | 0 | 17 |
| 0 | 17 | 1 | 18 | 1 | 18 | 1 | 19 | 1 | 17 | 1 | |
15 | 18 |
| 0 | 18 | 0 | 17 |
| 0 | 17 | 0 | 18 |
| 0 | 18 | 0 | 19 | 0 | 17 | 0 |
16 | 18 |
| 0 | 17 | 1 | 16 |
| 1 | 17 | 0 | 18 | 0 | 18 | 0 | 18 | 1 | 17 | 0 | |
17 | 18 |
| 0 | 17 | 0 | 16 |
| 0 | 17 | 0 | 18 |
| 0 | 18 | 0 | 18 | 0 | 17 | 0 |
18 | 18 |
| 0 | 17 | 0 | 16 |
| 0 | 17 | 0 | 18 | 0 | 17 | 1 | 18 | 0 | 17 | 0 | |
19 | 18 |
| 0 | 17 | 0 | 16 |
| 0 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 |
20 | 17 |
| 1 | 17 | 0 | 16 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 1 | |
21 | 17 |
| 0 | 17 | 0 | 15 |
| 1 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 16 | 0 |
22 | 17 |
| 0 | 17 | 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 0 | |
23 | 17 |
| 0 | 16 | 1 | 15 |
| 0 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 16 | 0 |
24 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 0 | |
25 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 16 | 0 |
26 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 0 | |
27 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 16 | 0 |
28 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 0 | |
29 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 16 | 0 |
30 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 0 | |
31 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 16 | 0 |
32 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 0 | |
33 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 |
| 0 | 17 | 0 | 18 | 0 | 16 | 0 |
34 | 17 |
| 0 | 16 | 0 | 15 |
| 0 | 17 | 0 | 18 | 0 | 17 | 0 | 18 | 0 | 16 | 0 |
Table 21 (continued) Number of Living and Dead Embryo/Larvae/Fish and Number of Abnormalities Observed During the Test
Nominal Concentrations (mg/L) |
|
|
| 4.3 |
|
|
|
|
|
| 10 |
|
| |||||||
Parallels |
| 1 |
| 2 |
|
|
| 3 |
| 4 |
| 1 |
| 2 |
| 3 |
|
| 4 | |
Days | S |
| M | S | M |
| S |
| M | S | M | S | M | S | M | S | M | S | M | |
0 | 25 |
| 0 | 25 | 0 |
| 25 |
| 0 | 25 | 0 | 25 |
| 0 | 25 | 0 | 25 | 0 | 25 | 0 |
1 | 20 |
| 5 | 23 | 2 |
| 20 |
| 5 | 19 | 6 | 20 |
| 5 | 24 | 1 | 22 | 3 | 24 | 1 |
2 | 20 |
| 0 | 23 (1) | 0 |
| 20 |
| 0 | 19 (2) | 0 | 20 |
| 0 | 24 (1) | 0 | 22 (1) | 0 | 24 | 0 |
3 | 20 |
| 0 | 22 | 1 |
| 19 |
| 1 | 19 | 0 | 20 |
| 0 | 24 (1) | 0 | 21 | 1 | 24 | 0 |
4 | 20 (1) |
| 0 | 22 | 0 |
| 19 |
| 0 | 19 (1) | 0 | 19 (1) |
| 1 | 24 | 0 | 21 | 0 | 24 | 0 |
5 | 19 |
| 1 | 22 | 0 |
| 19 |
| 0 | 18 | 1 | 19 (1) |
| 0 | 23 | 1 | 21 | 0 | 24 | 0 |
6 | 19 |
| 0 | 20 | 2 |
| 18 |
| 1 | 18 | 0 | 18 | 1 | 22 | 1 | 21 | 0 | 22 | 2 | |
7 | 19 |
| 0 | 19 | 1 |
| 18 |
| 0 | 18 | 0 | 17 |
| 1 | 22 | 0 | 19 | 2 | 22 | 0 |
8 | 19 |
| 0 | 19 | 0 |
| 18 |
| 0 | 18 | 0 | 17 | 0 | 22 | 0 | 19 | 0 | 22 | 0 | |
9 | 19 |
| 0 | 19 | 0 |
| 18 |
| 0 | 18 | 0 | 17 |
| 0 | 22 | 0 | 19 | 0 | 22 | 0 |
10 | 18 |
| 1 | 19 | 0 |
| 18 |
| 0 | 18 | 0 | 17 | 0 | 22 | 0 | 19 | 0 | 22 | 0 | |
11 | 18 |
| 0 | 19 | 0 |
| 18 |
| 0 | 18 | 0 | 17 |
| 0 | 22 | 0 | 19 | 0 | 22 | 0 |
12 | 18 |
| 0 | 19 | 0 |
| 18 |
| 0 | 18 | 0 | 17 | 0 | 21 | 1 | 19 | 0 | 22 | 0 | |
13 | 18 |
| 0 | 19 | 0 |
| 18 |
| 0 | 18 | 0 | 17 |
| 0 | 21 | 0 | 19 | 0 | 22 | 0 |
14 | 17 |
| 1 | 18 | 1 |
| 17 |
| 1 | 17 | 1 | 17 | 0 | 20 | 1 | 18 | 1 | 21 | 1 | |
15 | 17 |
| 0 | 18 | 0 |
| 17 |
| 0 | 17 | 0 | 17 |
| 0 | 20 | 0 | 18 | 0 | 21 | 0 |
16 | 17 |
| 0 | 18 | 0 |
| 17 |
| 0 | 17 | 0 | 17 | 0 | 19 | 1 | 18 | 0 | 20 | 1 | |
17 | 17 (1) |
| 0 | 18 (1) | 0 |
| 17 |
| 0 | 17 | 0 | 16 |
| 1 | 19 | 0 | 17 | 1 | 19 | 1 |
18 | 16 (1) |
| 1 | 18 (1) | 0 |
| 17 |
| 0 | 17 | 0 | 16 | 0 | 19 | 0 | 17 | 0 | 19 | 0 | |
19 | 16 (1) |
| 0 | 18 (1) | 0 |
| 17 |
| 0 | 17 | 0 | 16 (1) |
| 0 | 19 | 0 | 17 | 0 | 19 | 0 |
20 | 15 |
| 1 | 17 | 1 |
| 17 |
| 0 | 17 | 0 | 16 (1) | 0 | 19 | 0 | 17 | 0 | 19 | 0 | |
21 | 15 |
| 0 | 17 | 0 |
| 17 |
| 0 | 17 | 0 | 15 |
| 1 | 19 | 0 | 17 | 0 | 18 | 1 |
22 | 15 |
| 0 | 16 | 1 |
| 17 |
| 0 | 17 | 0 | 15 | 0 | 19 | 0 | 17 | 0 | 18 | 0 | |
23 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 17 | 0 | 15 |
| 0 | 19 | 0 | 17 | 0 | 18 | 0 |
24 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 16 | 1 | 15 | 0 | 19 | 0 | 17 | 0 | 18 | 0 | |
25 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 1 | 15 |
| 0 | 19 | 0 | 17 | 0 | 18 | 0 |
26 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 | 0 | 19 | 0 | 17 | 0 | 18 | 0 | |
27 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 |
| 0 | 19 | 0 | 17 | 0 | 18 | 0 |
28 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 | 0 | 19 | 0 | 17 | 0 | 18 | 0 | |
29 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 |
| 0 | 19 | 0 | 17 | 0 | 18 | 0 |
30 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 | 0 | 19 | 0 | 17 | 0 | 18 | 0 | |
31 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 |
| 0 | 19 | 0 | 17 | 0 | 18 | 0 |
32 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 | 0 | 19 | 0 | 17 | 0 | 18 | 0 | |
33 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 |
| 0 | 19 | 0 | 17 | 0 | 18 | 0 |
34 | 15 |
| 0 | 16 | 0 |
| 17 |
| 0 | 15 | 0 | 15 | 0 | 19 | 0 | 17 | 0 | 18 | 0 |
Table 22 Body Weight Data
Nominal concentrations | Parallels | Number of fish | Total body weighta | Mean body weightb |
(mg/L) |
|
| (g) | (g) |
Control | 1 | 15 | 0.262 | 0.0175 |
| 2 | 15 | 0.356 | 0.0237 |
| 3 | 19 | 0.404 | 0.0213 |
| 4 | 15 | 0.381 | 0.0254 |
| Mean | - | 0.351 | 0.0220 |
0.36 | 1 | 17 | 0.372 | 0.0219 |
| 2 | 17 | 0.330 | 0.0194 |
| 3 | 15 | 0.325 | 0.0217 |
| 4 | 15 | 0.384 | 0.0256 |
| Mean | - | 0.353 | 0.0221 |
0.82 | 1 | 17 | 0.371 | 0.0218 |
| 2 | 16 | 0.330 | 0.0206 |
| 3 | 15 | 0.308 | 0.0205 |
| 4 | 17 | 0.377 | 0.0222 |
| Mean | - | 0.347 | 0.0213 |
1.9 | 1 | 18 | 0.384 | 0.0213 |
| 2 | 17 | 0.384 | 0.0226 |
| 3 | 18 | 0.395 | 0.0219 |
| 4 | 16 | 0.370 | 0.0231 |
| Mean | - | 0.383 | 0.0222 |
4.3 | 1 | 15 | 0.292 | 0.0195 |
| 2 | 16 | 0.331 | 0.0207 |
| 3 | 17 | 0.403 | 0.0237 |
| 4 | 15 | 0.341 | 0.0227 |
| Mean | - | 0.342 | 0.0216 |
10 | 1 | 15 | 0.331 | 0.0221 |
| 2 | 19 | 0.494 | 0.0260 |
| 3 | 17 | 0.325 | 0.0191 |
| 4 | 18 | 0.371 | 0.0206 |
| Mean | - | 0.380 | 0.0219 |
a = Total body weights were measured on a replicate basis. b = Mean body weights were calculated for the surviving individual fish.
Table 23 Body Length Data
Nominal Concentrations (mg/L) |
| Control |
|
| 0.36 |
|
| 0.82 |
|
| 1.9 |
|
|
|
| 4.3 |
|
|
| 10 |
| |||
Parallels | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 |
1.75 1.80 1.80 1.60 1.50 1.65 1.65 1.45 1.50 1.30 1.45 1.25 1.25 | 1.70 1.75 1.45 1.55 1.75 1.50 1.60 1.60 1.40 1.75 1.55 1.75 1.25 | 1.45 1.60 1.50 1.60 1.35 1.80 1.55 1.45 1.55 1.55 1.05 1.50 1.55 | 1.50 1.60 1.85 1.60 1.65 1.60 1.60 1.85 1.60 1.60 1.55 1.25 1.10 | 1.60 1.50 1.55 1.60 1.45 1.55 1.55 1.60 1.60 1.65 1.45 1.50 1.30 | 1.45 1.50 1.60 1.40 1.45 1.40 1.40 1.50 1.45 1.55 1.45 1.35 1.35 | 1.60 1.45 1.50 1.45 1.50 1.50 1.55 1.40 1.35 1.45 1.85 1.65 1.45 | 1.35 1.60 1.65 1.60 1.60 1.60 1.60 1.55 1.60 1.70 1.55 1.55 1.45 | 1.50 1.45 1.45 1.50 1.60 1.65 1.60 1.55 1.50 1.55 1.50 1.55 1.60 | 1.35 1.75 1.65 1.35 1.60 1.45 1.45 1.55 1.50 1.60 1.25 1.45 1.35 | 1.20 1.55 1.40 1.35 1.20 1.45 1.40 1.80 1.55 1.65 1.60 1.25 1.85 | 1.65 1.35 1.50 1.55 1.40 1.45 1.60 1.60 1.45 1.85 1.60 1.20 1.35 | 1.20 1.65 1.45 1.30 1.20 1.40 1.45 1.55 1.65 1.60 1.60 1.55 1.30 | 1.70 1.60 1.75 1.50 1.75 1.65 1.60 1.45 1.65 1.55 1.35 1.50 1.45 | 1.65 1.35 1.85 1.60 1.45 1.35 1.50 1.75 1.50 1.35 1.45 1.35 1.35 | 1.65 1.50 1.70 1.45 1.60 1.50 1.60 1.60 1.60 1.80 1.35 1.50 1.50 | 1.10 1.60 1.55 1.50 1.60 1.55 1.50 1.70 1.75 1.60 1.25 1.45 1.40 | 1.65 1.45 1.35 1.35 1.75 1.60 1.55 1.55 1.40 1.25 1.80 1.30 1.40 | 1.60 1.65 1.50 1.50 1.60 1.55 1.20 1.65 1.60 1.45 1.45 1.45 1.50 | 1.45 1.45 1.50 1.55 1.40 1.45 1.45 1.65 1.60 1.55 1.55 1.60 1.60 | 1.70 1.65 1.60 1.75 1.45 1.50 1.70 1.30 1.50 1.50 1.50 1.75 1.70 | 1.25 1.60 1.45 1.50 1.60 1.30 1.45 1.45 1.45 1.55 1.50 1.35 1.60 | 1.60 1.50 1.45 1.25 1.45 1.55 1.50 1.60 1.50 1.40 1.45 1.55 1.70 | 1.70 1.55 1.45 1.60 1.50 1.60 1.45 1.45 1.40 1.30 1.30 1.15 1.60 | |
| 1.35 | 1.45 | 1.60 | 1.45 | 1.50 | 1.40 | 1.45 | 1.05 | 1.65 | 1.50 | 1.65 | 1.45 | 1.30 | 1.20 | 1.45 | 1.55 | 1.65 | 1.55 | 1.75 | 1.60 | 1.65 | 1.55 | 1.75 | 1.40 |
| 1.30 | 1.30 | 1.55 | 1.20 | 1.25 | 1.25 | 1.50 | 1.50 | 1.45 | 1.45 | 1.60 | 1.35 | 1.50 | 1.35 | 1.35 | 1.50 | 1.60 | 1.55 | 1.50 | 1.40 | 1.45 | 1.50 | 1.65 | 1.35 |
|
|
| 1.60 |
| 1.30 | 1.45 |
|
| 1.35 | 1.60 |
| 1.50 | 1.40 | 1.45 | 1.50 | 1.35 |
| 1.65 | 1.30 |
|
| 1.35 | 1.40 | 1.45 |
|
|
| 1.45 |
| 1.35 | 1.50 |
|
| 1.50 |
|
| 1.35 | 1.35 | 1.25 | 1.35 |
|
|
| 1.35 |
|
| 1.35 | 1.40 | 1.45 |
|
|
| 1.45 |
|
|
|
|
|
|
|
|
| 1.30 |
| 1.70 |
|
|
|
|
|
| 1.65 |
| 1.20 |
|
|
| 1.45 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| 1.80 |
|
|
Mean | 1.51 | 1.56 | 1.51 | 1.53 | 1.49 | 1.44 | 1.51 | 1.53 | 1.53 | 1.49 | 1.50 | 1.48 | 1.43 | 1.51 | 1.49 | 1.55 | 1.52 | 1.51 | 1.51 | 1.52 | 1.58 | 1.49 | 1.51 | 1.44 |
SD | 0.19 | 0.17 | 0.15 | 0.21 | 0.12 | 0.08 | 0.12 | 0.16 | 0.08 | 0.13 | 0.20 | 0.15 | 0.15 | 0.16 | 0.16 | 0.12 | 0.17 | 0.16 | 0.14 | 0.08 | 0.13 | 0.13 | 0.12 | 0.14 |
Group mean | 1.52 |
|
|
| 1.49 |
|
|
| 1.50 |
|
|
| 1.49 |
|
|
| 1.51 |
|
|
| 1.50 |
|
|
|
Number of fish measured | 15 | 15 | 19 | 15 | 17 | 17 | 15 | 15 | 17 | 16 | 15 | 17 | 18 | 17 | 18 | 16 | 15 | 16 | 17 | 15 | 15 | 19 | 17 | 18 |
Table 24 Feeding of Test Animals (Larvae/Juvenile)
Days | 0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 |
DF mg/day/test aquarium |
| No food supplied |
| 12.5 |
|
|
|
|
| 25 |
|
|
|
|
| |||
AN |
|
|
|
| N | o live food supplied |
|
|
|
|
|
|
| Not specified amount* |
|
Days | 18 | 19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 | 27 | 28 | 29 | 30 | 31 | 32 | 33 | 34 |
DF mg/day/test aquarium |
| 25 |
|
| 12.5 |
|
|
|
| No dry food |
|
|
|
| No food supplied | ||
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| |||
AN |
|
|
|
|
|
| Not specified amount* |
|
|
|
|
|
|
|
|
DF = Dry food (Sera micron Nature)
AN = Artemia naupli
* Both DF and AN were supplied ad libitum (means supplying excessive amount of food at any time of feeding). No amount of artemia nauplies were determined in the “suspension” supplied to each test chambers in same volume.
Description of key information
Under the conditions of this fish early life stage toxicity test, the
obtained results showed that the test item 1-ethoxy-2-(2-
methoxyethoxy)ethane had no significant lethal or sub-lethal effects
on early life stages of Zebrafish (Danio rerio) up to at least the
highest tested concentration of 10 mg/L. Accordingly, the test item
1-ethoxy-2-(2-methoxyethoxy)ethane had no chronic effects on
Zebrafish (Danio rerio) up to at least the tested maximum
concentration of 10 mg/L.
Table 1. Summary of Biological Endpoints
Endpoints | 1-ethoxy-2-(2-methoxyethoxy)ethane concentration (mg/L) |
Hatchability NOEC | ≥ 10 |
Hatchability LOEC | > 10 |
Post-hatch survival NOEC | ≥ 10 |
Post-hatch survival LOEC | > 10 |
Body weight (terminal) NOEC | ≥ 10 |
Body weight (terminal) LOEC | > 10 |
Body length (terminal) NOEC | ≥ 10 |
Body length (terminal) LOEC | > 10 |
34 day NOEC | ≥ 10 |
34 day LOEC | > 10 |
34 day ECx | > 10 |
Key value for chemical safety assessment
Fresh water fish
Fresh water fish
- Dose descriptor:
- NOEC
- Effect concentration:
- >= 10 mg/L
Additional information
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