Registration Dossier
Registration Dossier
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 267-636-0 | CAS number: 67905-17-3
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:
- 16/04/2020 to 08/02/2021
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: test concentration 1.00, 0.30, 0.10, 0.03 and 0.01 mg/L
- Sampling method: The test item in an amount of 30 mg and a volume of 300 µL DMSO quantitatively transferred into a volumetric flask and filled up to 3000 mL with the reconstituted water
- Sample storage conditions before analysis: samples were analysed immediately after preperation - Vehicle:
- yes
- Remarks:
- Acetone
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: test media ( reconstituted water), solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): DMSO
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): 0.01ml/L - Test organisms (species):
- Danio rerio (previous name: Brachydanio rerio)
- Details on test organisms:
- TEST ORGANISM
- Common name: Zebra fish
- Source: Tharun aqua fish farm, India
METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish (i.e. of females used to provide required number of eggs): Zebra fishes of both sex of ratio (2 male : 1female)
- Method of collection of fertilised eggs: Eggs were collected from breeding groups, mixed and randomly selected. Spawning traps were placed in the breeding tank for the collection of eggs. To prevent predation of eggs by adult zebra fish, the spawn traps were covered with inert wire mesh of size (2±0.5 mm).
- Subsequent handling of eggs: the spawn traps with the collected eggs were carefully removed. Eggs were rinsed with reconstituted water after collection from spawning traps
POST-HATCH FEEDING
- Start date: 5dpf
- Type/source of feed: 200 rotifer/ml and with paramecium ad libitum
- Amount given: ad libitum
- Frequency of feeding: daily - Test type:
- semi-static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 30 d
- Remarks on exposure duration:
- Total duration 32 days
- Hardness:
- Control: 190–220 mg/L (as CaCO3) / Solvent control:190 – 220 mg/L (as CaCO3)
- Test temperature:
- 24.6-26 °C
- pH:
- Control: 7.2–7.8 mg/L / Solvent control: 7.2–7.8 mg/L /Treatment: 7.1–7.8 mg/L
- Dissolved oxygen:
- Control: 68–78% / Solvent control: 68–78% /
Treatment: 68–78 % - Salinity:
- Not applicable
- Nominal and measured concentrations:
- Nominal concentration: 0.01, 0.03, 0.10, 0.30 and 1.00 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 7 litre of aquaria tank
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume:
- Aeration: no aeration
- Renewal requency:
- No. of fertilized eggs/embryos per vessel:
- No. of vessels per concentration (replicates):
- No. of vessels per control (replicates):
- No. of vessels per vehicle control (replicates):
- Biomass loading rate:
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Salinity:
- Culture medium different from test medium:
- Intervals of water quality measurement:
OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod:
- Light intensity:
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
VEHICLE CONTROL PERFORMED: yes/no
RANGE-FINDING STUDY
- Test concentrations:
- Results used to determine the conditions for the definitive study:
POST-HATCH DETAILS
- Begin of post-hatch period:
- No. of hatched eggs (alevins)/treatment released to the test chamber:
- Release of alevins from incubation cups to test chamber on day no.:
FERTILIZATION SUCCESS STUDY
- Number of eggs used:
- Removal of eggs to check the embryonic development on day no.: - Reference substance (positive control):
- not required
- Duration:
- 30 d
- Dose descriptor:
- LC50
- Effect conc.:
- 0.064 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Remarks on result:
- other: 0.064±0.01 mg/L
- Duration:
- 30 d
- Dose descriptor:
- NOEC
- Effect conc.:
- < 0.01 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Duration:
- 30 d
- Dose descriptor:
- LOEC
- Effect conc.:
- < 0.01 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Duration:
- 30 d
- Dose descriptor:
- EC50
- Effect conc.:
- 0.34 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- fertility
- Remarks on result:
- other: 0.34±0.047 mg/l
- Duration:
- 30 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.01 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- fertility
- Duration:
- 30 d
- Dose descriptor:
- LOEC
- Effect conc.:
- 0.03 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- fertility
- Duration:
- 30 d
- Dose descriptor:
- EC50
- Effect conc.:
- 0.31 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: survival larvae
- Remarks on result:
- other: 0.31±0.06 mg/L
- Details on results:
- - Mortality/survival at embryo, larval, juvenile, and adult stages:
LC50 mortality 0.064±0.01 EC50fertility 0.34±0.047 EC50survival 0.31±0.06
NOECmortality <0.01 NOECfertility 0.01 NOEC survival <0.01
LOECmortality <0.01 LOECfertility 0.03 LOEC survival <0.01
- Days to hatch or time to release of young: Hatching success in the control and solvent control group was 93.42 and 94.67% respectively, The NOEC and LOEC for hatching success were determined as 0.01 and 0.03 mg/L, respectively.First egg hatch in treatment, control and solvent control group occurred in the 24-hour period between the Day 2 and Day 3 pre-hatch observation timepoints
- Numbers hatched, : 76 embryos hatched in control and 75 embbryos hatched in solvent control
- Observations on body length and weight of young and/or exposed parents at one or more time periods:
Test concentrations mg/L: mean length (mm) mean dry weight (mg)
1 8.88 21.50
0.30 9.87 23.03
0.10 10.07 23.5
0.03 11.32 25.62
0.01 11.46 26.29
Solvent control 11.97 27.37
Control 11.93 27.53
other results are included in tables sections - Reported statistics and error estimates:
- The endpoint values were determined within a given exposure period by using a Probit analysis in the NCSS (Number Cruncher Statistical System) and one-way ANOVA using Graphpad Prism 8.0.
- Validity criteria fulfilled:
- yes
- Remarks:
- Water temp was maintained at 26 ± 1.5 °C throughout the exposure. The DO in all groups was between 68–78 % of saturation. Hatching rate in control is 93.42% and solvent control is 94.67. Larvae survival in control, solvent control was 95.0 and 93.75%.
- Conclusions:
- The median effects of test chemical on mortality, hatching success and fish larvae survival Danio rerio is 0.064±0.01, 0.34±0.047 and 0.31±0.06 mg/l respectively
- Executive summary:
The aim of the study was todetermine the EC50, LOEC and NOECof test chemical on the growth and development of embryos and larvae of the freshwater fish species zebrafish (Danio rerio) in a Fish Early-Life Stage (ELS) Toxicity Test. The study was conducted according to the OECD guideline No. 210, Adopted 26thJuly, 2013.
The test was conducted with a semi-static design with renewal once 48 h.
Concentrated stock solutions and test media were analysed during the ELS test. Based on the range finding result the definitive test was conducted with five test concentrations of 0.01, 0.03, 0.10, 0.30 and 1.00 mg/L in terms of test chemical with a conversion factor of 3.2. Both control and solvent control were included in the test design. Each concentration was divided into four replicate with 20 eggs each and including the controls.
The cumulative mortality of the control and solvent control group at 30 days post-hatch was 11.25 and 11.25 %, whereas corrected cumulative mortality of embryos, larvae and juvenile fish after at the end of exposure to test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg /Lwas 12.68, 28.45, 49.30, 67.61and 92.96%,respectively.
Hatching success in the control and solvent control group was 93.42 and 94.67% respectively, satisfying the validation criterion for hatching success (70%),whereas in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas 91.18, 87.93, 67.92, 53.49and 25.00%,respectively. Fertility reduction in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas3.69, 7.12, 28.26, 43.50 and 73.59%,respectively in comparison with the solvent control group.The NOEC and LOEC for hatching success were determined as 0.01 and 0.03 mg/L, respectively.
Larval survival until Day 30 post-hatch in the control and solvent control group was 95.0 and 93.75% thereby exceeding and satisfying the validity criteria for post-hatch survival (>75%).whereas in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas85.0, 72.50, 66.25, 53.75 and 25.0%,respectively. Larval survival reduction in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas 9.33, 22.67, 29.33, 42.67 and 73.33%,respectively in comparison with the solvent control group.In terms of concentrations, the NOEC and LOEC for post-hatch larval survival untilDay 30 were both considered to be equal to or less than 0.01 mg/L.
For fish total lengths, the NOEC and LOEC determined on Day 30 post-hatch were 0.01 and 0.03mg/L, respectively. For fish dry weights, the NOEC and LOEC determined on Day 30 post-hatch were 0.01 and 0.03 mg/L, respectively.
The concentrations of test chemical was determined using a validated liquid chromatographic method. Samples of each fresh test substance concentration, control and solvent control collected at exposure initiation and at renewals, and samples of each spent test substance concentration, control and solvent control collected at each renewal and at exposure termination were chemically determined.
In fresh samples of the test substance concentrations, the determined concentrations of test chemical was between 99.3 and 101.7%, concentrations, respectively. The results confirm that the test substance concentrations were within the acceptability range. In spent samples of the test substance concentrations, the determined concentrations of test chemical was between 96.1 and 96.9% of the concentrations, respectively.
All validity criteria were satisfied during the test, therefore the test was considered to be valid.
Results:The effects of test chemical on mortality, hatching success and fish larvae survivalDanio rerioin the definitive test are summarized in Table below.
LC50mortality
0.064±0.01
EC50fertility
0.34±0.047
EC50surviving
0.31±0.06
NOECmortality
<0.01
NOECfertility
0.01
NOECsurviving
<0.01
LOECmortality
<0.01
LOECfertility
0.03
LOECsurviving
<0.01
Thus based on the results obtained chemical can be categorised as into chronic category 1 as per CLP classification criteria.
Reference
Individual length (cm) and dry weight (mg) of fish (Control)– Range Finding Test
S. No. |
Control |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
11 |
12 |
12 |
11 |
26 |
28 |
28 |
24 |
|
2 |
13 |
14 |
13 |
13 |
31 |
31 |
30 |
30 |
|
3 |
12 |
13 |
12 |
13 |
28 |
29 |
28 |
28 |
|
4 |
12 |
12 |
12 |
14 |
27 |
28 |
26 |
31 |
|
5 |
13 |
11 |
11 |
12 |
29 |
24 |
22 |
28 |
|
6 |
13 |
12 |
12 |
11 |
30 |
28 |
24 |
25 |
|
7 |
12 |
11 |
13 |
12 |
25 |
26 |
29 |
27 |
|
8 |
12 |
13 |
12 |
12 |
26 |
29 |
27 |
27 |
|
9 |
13 |
13 |
11 |
13 |
28 |
30 |
26 |
29 |
|
10 |
13 |
12 |
11 |
11 |
29 |
27 |
24 |
25 |
|
11 |
11 |
12 |
11 |
12 |
24 |
28 |
24 |
27 |
|
12 |
12 |
11 |
12 |
12 |
25 |
25 |
28 |
25 |
|
13 |
12 |
12 |
12 |
13 |
26 |
28 |
29 |
29 |
|
14 |
13 |
11 |
13 |
11 |
26 |
25 |
30 |
24 |
|
15 |
14 |
13 |
13 |
13 |
32 |
30 |
31 |
30 |
|
16 |
11 |
12 |
12 |
12 |
24 |
28 |
28 |
28 |
|
17 |
12 |
12 |
11 |
11 |
26 |
26 |
24 |
24 |
|
18 |
12 |
11 |
13 |
12 |
27 |
24 |
29 |
27 |
|
19 |
11 |
- |
12 |
13 |
23 |
- |
27 |
29 |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
12.21 |
12.06 |
12.00 |
12.16 |
26.95 |
27.44 |
27.05 |
27.21 |
|
Total mean |
12.11 |
27.16 |
|||||||
Minimum |
11 |
22 |
|||||||
Maximum |
14 |
32 |
|||||||
Individual SD |
0.85 |
0.87 |
0.75 |
0.90 |
2.44 |
2.06 |
2.50 |
2.20 |
|
Total SD |
0.84 |
2.30 |
|||||||
R- Replicate; SD – Standard Deviation
Individual length (cm) and dry weight (mg) of fish (Solvent Control)– Range Finding Test
S. No. |
Control |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
12 |
11 |
12 |
12 |
27 |
26 |
28 |
25 |
|
2 |
11 |
13 |
11 |
13 |
26 |
30 |
25 |
28 |
|
3 |
12 |
12 |
12 |
12 |
28 |
27 |
26 |
25 |
|
4 |
13 |
11 |
13 |
12 |
29 |
24 |
29 |
28 |
|
5 |
14 |
13 |
13 |
13 |
32 |
30 |
30 |
29 |
|
6 |
12 |
12 |
12 |
12 |
28 |
25 |
28 |
24 |
|
7 |
12 |
11 |
11 |
11 |
26 |
24 |
24 |
22 |
|
8 |
11 |
12 |
12 |
12 |
24 |
28 |
26 |
26 |
|
9 |
13 |
13 |
11 |
12 |
29 |
29 |
22 |
25 |
|
10 |
13 |
11 |
13 |
12 |
30 |
24 |
29 |
27 |
|
11 |
12 |
12 |
13 |
13 |
28 |
28 |
27 |
28 |
|
12 |
13 |
13 |
12 |
12 |
29 |
30 |
25 |
25 |
|
13 |
12 |
11 |
12 |
12 |
26 |
25 |
23 |
26 |
|
14 |
11 |
13 |
12 |
11 |
24 |
29 |
26 |
24 |
|
15 |
12 |
13 |
11 |
11 |
25 |
31 |
24 |
24 |
|
16 |
13 |
12 |
13 |
12 |
28 |
28 |
29 |
28 |
|
17 |
11 |
11 |
11 |
13 |
21 |
27 |
24 |
30 |
|
18 |
12 |
12 |
12 |
11 |
25 |
27 |
26 |
25 |
|
19 |
- |
11 |
- |
- |
- |
26 |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
12.17 |
11.95 |
12.00 |
12.00 |
26.94 |
27.26 |
26.17 |
26.06 |
|
Total mean |
12.03 |
26.61 |
|||||||
Minimum |
11 |
21 |
|||||||
Maximum |
14 |
32 |
|||||||
Individual SD |
0.86 |
0.85 |
0.77 |
0.69 |
2.60 |
2.23 |
2.31 |
2.10 |
|
Total SD |
0.79 |
2.31 |
|||||||
R- Replicate; SD – Standard Deviation
Individual length (cm) and dry weight (mg) of fish (0.01 mg/L)– Range Finding Test
S. No. |
Control |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
12 |
12 |
11 |
11 |
28 |
28 |
23 |
24 |
|
2 |
11 |
13 |
12 |
13 |
26 |
30 |
25 |
31 |
|
3 |
12 |
12 |
10 |
10 |
28 |
27 |
22 |
21 |
|
4 |
13 |
11 |
11 |
11 |
29 |
25 |
21 |
23 |
|
5 |
12 |
12 |
10 |
10 |
25 |
28 |
22 |
20 |
|
6 |
12 |
13 |
12 |
12 |
28 |
29 |
22 |
27 |
|
7 |
13 |
11 |
13 |
13 |
29 |
26 |
29 |
29 |
|
8 |
11 |
12 |
10 |
11 |
24 |
27 |
22 |
25 |
|
9 |
12 |
12 |
11 |
12 |
26 |
24 |
24 |
26 |
|
10 |
12 |
13 |
12 |
11 |
28 |
24 |
27 |
24 |
|
11 |
12 |
12 |
13 |
10 |
27 |
28 |
29 |
22 |
|
12 |
13 |
11 |
11 |
12 |
28 |
22 |
24 |
28 |
|
13 |
11 |
13 |
12 |
13 |
23 |
28 |
26 |
29 |
|
14 |
11 |
12 |
11 |
10 |
24 |
26 |
21 |
21 |
|
15 |
12 |
12 |
11 |
12 |
27 |
27 |
24 |
25 |
|
16 |
13 |
11 |
- |
11 |
28 |
24 |
- |
23 |
|
17 |
- |
11 |
- |
- |
- |
22 |
- |
- |
|
18 |
- |
- |
- |
- |
- |
- |
- |
- |
|
19 |
- |
- |
- |
- |
- |
- |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
12.00 |
11.94 |
11.33 |
11.38 |
26.75 |
26.18 |
24.07 |
24.88 |
|
Total mean |
11.66 |
25.47 |
|||||||
Minimum |
10 |
20 |
|||||||
Maximum |
13 |
31 |
|||||||
Individual SD |
0.73 |
0.75 |
0.98 |
1.09 |
1.88 |
2.35 |
2.66 |
3.24 |
|
Total SD |
0.89 |
2.53 |
|||||||
R- Replicate; SD – Standard Deviation
Individual length (cm) and dry weight (mg) of fish (0.10 mg/L)– Range Finding Test
S. No. |
Control |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
12 |
12 |
10 |
11 |
28 |
25 |
22 |
26 |
|
2 |
11 |
11 |
11 |
12 |
23 |
23 |
24 |
27 |
|
3 |
13 |
12 |
10 |
11 |
28 |
25 |
20 |
22 |
|
4 |
12 |
10 |
12 |
10 |
26 |
21 |
23 |
23 |
|
5 |
10 |
10 |
12 |
10 |
22 |
21 |
25 |
21 |
|
6 |
12 |
11 |
10 |
13 |
24 |
24 |
20 |
28 |
|
7 |
11 |
12 |
11 |
12 |
23 |
26 |
20 |
25 |
|
8 |
10 |
12 |
11 |
11 |
22 |
25 |
22 |
22 |
|
9 |
11 |
10 |
10 |
10 |
26 |
21 |
19 |
21 |
|
10 |
11 |
10 |
12 |
10 |
25 |
23 |
26 |
21 |
|
11 |
- |
11 |
10 |
9 |
- |
26 |
23 |
19 |
|
12 |
- |
- |
11 |
10 |
- |
- |
26 |
23 |
|
13 |
- |
- |
- |
12 |
- |
- |
- |
28 |
|
14 |
- |
- |
- |
- |
- |
- |
- |
- |
|
15 |
- |
- |
- |
- |
- |
- |
- |
- |
|
16 |
- |
- |
- |
- |
- |
- |
- |
- |
|
17 |
- |
- |
- |
- |
- |
- |
- |
- |
|
18 |
- |
- |
- |
- |
- |
- |
- |
- |
|
19 |
- |
- |
- |
- |
- |
- |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
11.30 |
11.00 |
10.83 |
10.85 |
24.70 |
23.64 |
22.50 |
23.54 |
|
Total mean |
10.99 |
23.59 |
|||||||
Minimum |
9 |
19 |
|||||||
Maximum |
13 |
28 |
|||||||
Individual SD |
0.95 |
0.89 |
0.83 |
1.14 |
2.26 |
1.96 |
2.43 |
2.96 |
|
Total SD |
0.96 |
2.40 |
|||||||
R- Replicate; SD – Standard Deviation
Individual length (cm) and dry weight (mg) of fish (1.00 mg/L)– Range Finding Test
S. No. |
Control |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
9 |
10 |
10 |
9 |
20 |
25 |
24 |
20 |
|
2 |
11 |
9 |
- |
8 |
25 |
22 |
- |
18 |
|
3 |
- |
- |
- |
9 |
- |
- |
- |
21 |
|
4 |
- |
- |
- |
- |
- |
- |
- |
|
|
5 |
- |
- |
- |
- |
- |
- |
- |
- |
|
6 |
- |
- |
- |
- |
- |
- |
- |
- |
|
7 |
- |
- |
- |
- |
- |
- |
- |
- |
|
8 |
- |
- |
- |
- |
- |
- |
- |
- |
|
9 |
- |
- |
- |
- |
- |
- |
- |
- |
|
10 |
- |
- |
- |
- |
- |
- |
- |
- |
|
11 |
- |
- |
- |
- |
- |
- |
- |
- |
|
12 |
- |
- |
- |
- |
- |
- |
- |
- |
|
13 |
- |
- |
- |
- |
- |
- |
- |
- |
|
14 |
- |
- |
- |
- |
- |
- |
- |
- |
|
15 |
- |
- |
- |
- |
- |
- |
- |
- |
|
16 |
- |
- |
- |
- |
- |
- |
- |
- |
|
17 |
- |
- |
- |
- |
- |
- |
- |
- |
|
18 |
- |
- |
- |
- |
- |
- |
- |
- |
|
19 |
- |
- |
- |
- |
- |
- |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
10.00 |
9.50 |
10.00 |
8.67 |
22.50 |
23.50 |
24.00 |
19.67 |
|
Total mean |
9.54 |
22.42 |
|||||||
Minimum |
8 |
18 |
|||||||
Maximum |
11 |
25 |
|||||||
Individual SD |
1.41 |
0.71 |
0.00 |
0.58 |
3.54 |
2.12 |
0.00 |
1.53 |
|
Total SD |
0.67 |
1.80 |
|||||||
R- Replicate; SD – Standard Deviation
10 Individual length (cm) and dry weight (mg) of fish (Control)– Definitive test
S. No. |
Control |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
12 |
13 |
11 |
12 |
27 |
31 |
26 |
26 |
|
2 |
13 |
11 |
11 |
14 |
30 |
26 |
25 |
32 |
|
3 |
11 |
12 |
13 |
12 |
26 |
26 |
31 |
27 |
|
4 |
12 |
13 |
12 |
11 |
28 |
29 |
29 |
25 |
|
5 |
13 |
13 |
11 |
11 |
32 |
31 |
27 |
25 |
|
6 |
11 |
11 |
11 |
12 |
25 |
26 |
26 |
28 |
|
7 |
12 |
11 |
14 |
12 |
28 |
25 |
32 |
27 |
|
8 |
12 |
12 |
12 |
13 |
27 |
26 |
28 |
31 |
|
9 |
11 |
11 |
12 |
11 |
25 |
26 |
29 |
25 |
|
10 |
13 |
13 |
11 |
13 |
30 |
29 |
25 |
29 |
|
11 |
11 |
11 |
12 |
11 |
26 |
25 |
27 |
25 |
|
12 |
12 |
12 |
13 |
12 |
29 |
27 |
31 |
27 |
|
13 |
11 |
11 |
13 |
12 |
25 |
25 |
30 |
26 |
|
14 |
12 |
12 |
11 |
12 |
28 |
28 |
25 |
26 |
|
15 |
11 |
12 |
12 |
11 |
26 |
29 |
27 |
26 |
|
16 |
12 |
12 |
11 |
13 |
28 |
28 |
26 |
29 |
|
17 |
14 |
11 |
- |
12 |
30 |
25 |
- |
27 |
|
18 |
11 |
13 |
- |
13 |
27 |
30 |
- |
31 |
|
19 |
- |
12 |
- |
- |
- |
29 |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
11.89 |
11.89 |
11.88 |
12.06 |
27.61 |
27.42 |
27.75 |
27.33 |
|
Total mean |
11.93 |
27.53 |
|||||||
Minimum |
11 |
25 |
|||||||
Maximum |
14 |
32 |
|||||||
Individual SD |
0.90 |
0.81 |
0.96 |
0.87 |
2.0 |
2.06 |
2.32 |
2.22 |
|
Total SD |
0.88 |
2.15 |
|||||||
R- Replicate; SD – Standard Deviation
Individual length and weight of fish (Solvent control) - Definitive test
S. No. |
Solvent control |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
12 |
12 |
12 |
13 |
27 |
27 |
27 |
30 |
|
2 |
13 |
11 |
13 |
11 |
30 |
25 |
28 |
26 |
|
3 |
12 |
12 |
12 |
14 |
28 |
29 |
26 |
32 |
|
4 |
12 |
13 |
11 |
12 |
26 |
31 |
25 |
27 |
|
5 |
13 |
11 |
13 |
11 |
29 |
26 |
29 |
26 |
|
6 |
11 |
11 |
12 |
12 |
25 |
25 |
25 |
28 |
|
7 |
11 |
11 |
11 |
12 |
26 |
25 |
25 |
27 |
|
8 |
11 |
13 |
12 |
13 |
25 |
29 |
26 |
29 |
|
9 |
12 |
13 |
14 |
13 |
26 |
30 |
31 |
31 |
|
10 |
12 |
11 |
11 |
11 |
27 |
26 |
26 |
26 |
|
11 |
13 |
12 |
12 |
12 |
29 |
27 |
26 |
26 |
|
12 |
12 |
11 |
13 |
13 |
27 |
25 |
29 |
29 |
|
13 |
14 |
12 |
11 |
11 |
32 |
27 |
25 |
25 |
|
14 |
11 |
11 |
11 |
11 |
26 |
26 |
26 |
26 |
|
15 |
13 |
12 |
12 |
12 |
30 |
28 |
28 |
26 |
|
16 |
12 |
11 |
11 |
11 |
27 |
25 |
25 |
26 |
|
17 |
11 |
13 |
12 |
13 |
26 |
32 |
27 |
31 |
|
18 |
- |
12 |
13 |
12 |
|
30 |
31 |
28 |
|
19 |
- |
- |
- |
- |
- |
- |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
12.06 |
11.78 |
12.00 |
12.06 |
27.41 |
27.39 |
26.94 |
27.72 |
|
Total mean |
11.97 |
27.37 |
|||||||
Minimum |
11 |
25 |
|||||||
Maximum |
14 |
32 |
|||||||
Individual SD |
0.90 |
0.81 |
0.91 |
0.94 |
1.97 |
2.28 |
1.98 |
2.14 |
|
Total SD |
0.89 |
2.09 |
|||||||
R- Replicate; SD – Standard Deviation
Individual length and weight of fish (0.03 mg/L Concentration)- Definitive test
S. No. |
0.03 mg/L Concentration |
|||||||||||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
|||||||||||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
|||||||||||
1 |
11 |
11 |
12 |
11 |
25 |
25 |
27 |
24 |
||||||||||
2 |
12 |
12 |
11 |
12 |
27 |
27 |
22 |
23 |
||||||||||
3 |
13 |
12 |
13 |
11 |
30 |
27 |
29 |
25 |
||||||||||
4 |
11 |
11 |
11 |
12 |
27 |
26 |
25 |
27 |
||||||||||
5 |
12 |
12 |
10 |
10 |
28 |
27 |
24 |
23 |
||||||||||
6 |
10 |
9 |
11 |
11 |
24 |
23 |
26 |
25 |
||||||||||
7 |
11 |
10 |
12 |
12 |
25 |
23 |
27 |
26 |
||||||||||
8 |
12 |
11 |
12 |
12 |
28 |
24 |
26 |
27 |
||||||||||
9 |
11 |
11 |
11 |
11 |
27 |
25 |
26 |
26 |
||||||||||
10 |
13 |
12 |
12 |
11 |
30 |
27 |
28 |
25 |
||||||||||
11 |
12 |
10 |
11 |
10 |
26 |
24 |
22 |
23 |
||||||||||
12 |
11 |
12 |
10 |
12 |
25 |
28 |
23 |
26 |
||||||||||
13 |
- |
11 |
- |
11 |
- |
23 |
- |
24 |
||||||||||
14 |
- |
12 |
- |
- |
- |
25 |
- |
- |
||||||||||
15 |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||
16 |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||
17 |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||
18 |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||
19 |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||
20 |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||
Individual mean |
11.58 |
11.14 |
11.33 |
11.23 |
26.83 |
25.29 |
25.42 |
24.92 |
||||||||||
Total mean |
11.32 |
25.62 |
||||||||||||||||
Minimum |
9 |
22 |
||||||||||||||||
Maximum |
13 |
30 |
||||||||||||||||
Individual SD |
0.90 |
0.95 |
0.89 |
0.73 |
1.95 |
1.73 |
2.27 |
1.44 |
||||||||||
Total SD |
0.87 |
1.85 |
||||||||||||||||
R- Replicate; SD – Standard Deviation
Individual length and weight of fish (0.10 mg/L Concentration) - Definitive test
S. No. |
0.1 mg/L Concentration |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
9 |
10 |
11 |
9 |
22 |
24 |
24 |
22 |
|
2 |
10 |
11 |
10 |
10 |
24 |
26 |
24 |
24 |
|
3 |
11 |
10 |
10 |
10 |
25 |
24 |
23 |
23 |
|
4 |
10 |
11 |
11 |
11 |
23 |
25 |
25 |
25 |
|
5 |
11 |
9 |
9 |
11 |
25 |
22 |
22 |
24 |
|
6 |
9 |
10 |
10 |
10 |
21 |
24 |
23 |
23 |
|
7 |
10 |
11 |
9 |
9 |
24 |
25 |
21 |
21 |
|
8 |
9 |
9 |
11 |
11 |
23 |
23 |
24 |
24 |
|
9 |
- |
10 |
10 |
10 |
- |
23 |
23 |
23 |
|
10 |
- |
- |
11 |
- |
- |
- |
25 |
- |
|
11 |
- |
- |
- |
- |
- |
- |
- |
- |
|
12 |
- |
- |
- |
- |
- |
- |
- |
- |
|
13 |
- |
- |
- |
- |
- |
- |
- |
- |
|
14 |
- |
- |
- |
- |
- |
- |
- |
- |
|
15 |
- |
- |
- |
- |
- |
- |
- |
- |
|
16 |
- |
- |
- |
- |
- |
- |
- |
- |
|
17 |
- |
- |
- |
- |
- |
- |
- |
- |
|
18 |
- |
- |
- |
- |
- |
- |
- |
- |
|
19 |
- |
- |
- |
- |
- |
- |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
9.88 |
10.11 |
10.20 |
10.11 |
23.38 |
24.00 |
23.40 |
23.22 |
|
Total mean |
10.07+ |
23.5+ |
|||||||
Minimum |
9 |
21 |
|||||||
Maximum |
11 |
26 |
|||||||
Individual SD |
0.83 |
0.78 |
0.79 |
0.78 |
1.41 |
1.22 |
1.26 |
1.20 |
|
Total SD |
0.80 |
1.27 |
|||||||
R- Replicate; SD – Standard Deviation
+: statistically significant difference between the solvent control and the treatment group at p < 0.05
Individual length and weight of fish (0.30 mg/L Concentration) - Definitive test
S. No. |
0.3 mg/L Concentration |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
10 |
9 |
10 |
10 |
24 |
21 |
24 |
22 |
|
2 |
9 |
10 |
9 |
11 |
22 |
24 |
22 |
24 |
|
3 |
10 |
10 |
11 |
10 |
24 |
23 |
25 |
23 |
|
4 |
11 |
11 |
10 |
9 |
26 |
25 |
24 |
22 |
|
5 |
10 |
9 |
10 |
9 |
23 |
22 |
23 |
21 |
|
6 |
9 |
10 |
- |
- |
21 |
22 |
- |
- |
|
7 |
10 |
- |
- |
- |
23 |
- |
- |
- |
|
8 |
- |
- |
- |
- |
- |
- |
- |
- |
|
9 |
- |
- |
- |
- |
- |
- |
- |
- |
|
10 |
- |
- |
- |
- |
- |
- |
- |
- |
|
11 |
- |
- |
- |
- |
- |
- |
- |
- |
|
12 |
- |
- |
- |
- |
- |
- |
- |
- |
|
13 |
- |
- |
- |
- |
- |
- |
- |
- |
|
14 |
- |
- |
- |
- |
- |
- |
- |
- |
|
15 |
- |
- |
- |
- |
- |
- |
- |
- |
|
16 |
- |
- |
- |
- |
- |
- |
- |
- |
|
17 |
- |
- |
- |
- |
- |
- |
- |
- |
|
18 |
- |
- |
- |
- |
- |
- |
- |
- |
|
19 |
- |
- |
- |
- |
- |
- |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
9.86 |
9.83 |
10.00 |
9.80 |
23.29 |
22.83 |
23.60 |
22.40 |
|
Total mean |
9.87+ |
23.03+ |
|||||||
Minimum |
9 |
21 |
|||||||
Maximum |
11 |
26 |
|||||||
Individual SD |
0.69 |
0.75 |
0.71 |
0.84 |
1.60 |
1.47 |
1.14 |
1.14 |
|
Total SD |
0.75 |
1.34 |
|||||||
R- Replicate; SD – Standard Deviation
+: statistically significant difference between the solvent control and the treatment group at p < 0.05
Individual length and weight of fish (1.00 mg/L Concentration) - Definitive test
S. No. |
1 mg/L Concentration |
||||||||
R1 |
R2 |
R3 |
R4 |
R1 |
R2 |
R3 |
R4 |
||
Length (mm) |
Length (mm) |
Length (mm) |
Length (mm) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
Dry Weight (mg) |
||
1 |
8 |
10 |
9 |
9 |
20 |
23 |
22 |
22 |
|
2 |
- |
9 |
- |
- |
- |
21 |
- |
- |
|
3 |
- |
- |
- |
- |
- |
- |
- |
- |
|
4 |
- |
- |
- |
- |
- |
- |
- |
- |
|
5 |
- |
- |
- |
- |
- |
- |
- |
- |
|
6 |
- |
- |
- |
- |
- |
- |
- |
- |
|
7 |
- |
- |
- |
- |
- |
- |
- |
- |
|
8 |
- |
- |
- |
- |
- |
- |
- |
- |
|
9 |
- |
- |
- |
- |
- |
- |
- |
- |
|
10 |
- |
- |
- |
- |
- |
- |
- |
- |
|
11 |
- |
- |
- |
- |
- |
- |
- |
- |
|
12 |
- |
- |
- |
- |
- |
- |
- |
- |
|
13 |
- |
- |
- |
- |
- |
- |
- |
- |
|
14 |
- |
- |
- |
- |
- |
- |
- |
- |
|
15 |
- |
- |
- |
- |
- |
- |
- |
- |
|
16 |
- |
- |
- |
- |
- |
- |
- |
- |
|
17 |
- |
- |
- |
- |
- |
- |
- |
- |
|
18 |
- |
- |
- |
- |
- |
- |
- |
- |
|
19 |
- |
- |
- |
- |
- |
- |
- |
- |
|
20 |
- |
- |
- |
- |
- |
- |
- |
- |
|
Individual mean |
8.00 |
9.50 |
9.0 |
9.00 |
20 |
22 |
22 |
22 |
|
Total mean |
8.88+ |
21.50+ |
|||||||
Minimum |
8 |
20 |
|||||||
Maximum |
10 |
23 |
|||||||
Individual SD |
0 |
0.71 |
0 |
0 |
0 |
1.41 |
0 |
0 |
|
Total SD |
0.36 |
0.71 |
|||||||
R- Replicate; SD – Standard Deviation
+: statistically significant difference between the solvent control and the treatment group at p < 0.05
10 The Physico-Chemical Parameters – Range Finding Test
Hours/Day |
Parameter |
Concentrations (mg/L) |
||||||
Control |
Solvent control |
0.01 |
0.10 |
1.00 |
10.00# |
20.00# |
||
0 hr (Day 0) (Fresh) |
pH |
7.7 |
7.7 |
7.8 |
7.8 |
7.8 |
7.7 |
7.8 |
Temperature(°C) |
25.7 |
25.8 |
25.6 |
25.6 |
25.8 |
25.5 |
25.6 |
|
Dissolved oxygen (%) |
78 |
78 |
79 |
78 |
78 |
78 |
76 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
48 hr (Day 2) (Spent) |
pH |
7.5 |
7.5 |
7.5 |
7.5 |
7.6 |
7.2 |
7.1 |
Temperature(°C) |
24.9 |
24.8 |
25.1 |
25 |
24.9 |
25.1 |
25.2 |
|
Dissolved oxygen (%) |
70 |
71 |
71 |
71 |
71 |
70 |
70 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
48 hr (Day 2) (Fresh) |
pH |
7.8 |
7.6 |
7.6 |
7.7 |
7.7 |
- |
- |
Temperature(°C) |
25.7 |
25.6 |
25.3 |
25.4 |
25.2 |
- |
- |
|
Dissolved oxygen (%) |
76 |
76 |
76 |
76 |
76 |
- |
- |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
96 hr (Day 4) Spent |
pH |
7.3 |
7.2 |
7.3 |
7.3 |
7.3 |
- |
- |
Temperature(°C) |
25.6 |
25.5 |
25.6 |
25.5 |
25.7 |
- |
- |
|
Dissolved oxygen (%) |
71 |
71 |
71 |
70 |
70 |
- |
- |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
- |
- |
|
96 hr (Day 4) Fresh |
pH |
7.6 |
7.7 |
7.7 |
7.6 |
7.7 |
- |
- |
Temperature(°C) |
25.6 |
25.8 |
25.8 |
25.6 |
25.7 |
- |
- |
|
Dissolved oxygen (%) |
75 |
76 |
76 |
75 |
76 |
- |
- |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Day 6 (Spent) |
pH |
7.5 |
7.4 |
7.4 |
7.5 |
7.5 |
- |
- |
Temperature(°C) |
24.9 |
25 |
25 |
25.1 |
25 |
- |
- |
|
Dissolved oxygen (%) |
68 |
69 |
68 |
68 |
68 |
- |
- |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Day 6 (Fresh) |
pH |
7.8 |
7.8 |
7.7 |
7.7 |
7.8 |
- |
- |
Temperature(°C) |
25.3 |
25.4 |
25.4 |
25.6 |
25.4 |
- |
- |
|
Dissolved oxygen (%) |
72 |
73 |
72 |
72 |
73 |
- |
- |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
Day 8 (Spent) |
pH |
7.4 |
7.5 |
7.4 |
7.4 |
7.4 |
- |
- |
Temperature(°C) |
25.8 |
25.6 |
25.8 |
25.6 |
25.7 |
- |
- |
|
Dissolved oxygen (%) |
70 |
69 |
69 |
69 |
68 |
- |
- |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
# - due to dead all embryos in the test item concentration of 10 and 20 mg/L after 48 h, the physico-chemical parameters were not determined at remaining days of the exposure period.
(Continued) The Physico-Chemical Parameters – Range Finding Test
Day |
Parameter |
Concentrations (mg/L) |
||||
Control |
Solvent control |
0.01 |
0.10 |
1.00 |
||
Day 8 (Fresh) |
pH |
7.8 |
7.6 |
7.6 |
7.6 |
7.7 |
Temperature(°C) |
25.7 |
25.8 |
25.6 |
25.8 |
25.8 |
|
Dissolved oxygen (%) |
75 |
76 |
75 |
75 |
75 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 10 (Spent) |
pH |
7.2 |
7.2 |
7.3 |
7.3 |
7.2 |
Temperature(°C) |
25.5 |
25.6 |
25.5 |
25.6 |
25.6 |
|
Dissolved oxygen (%) |
70 |
71 |
71 |
70 |
70 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 10 (Fresh) |
pH |
7.7 |
7.7 |
7.7 |
7.7 |
7.6 |
Temperature(°C) |
25.8 |
25.6 |
25.6 |
25.5 |
25.6 |
|
Dissolved oxygen (%) |
77 |
78 |
78 |
77 |
78 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 12 (Spent) |
pH |
7.3 |
7.2 |
7.2 |
7.3 |
7.3 |
Temperature(°C) |
25.9 |
25.7 |
25.9 |
25.8 |
25.8 |
|
Dissolved oxygen (%) |
72 |
73 |
72 |
72 |
73 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
|
Day 12 (Fresh) |
pH |
7.5 |
7.6 |
7.5 |
7.5 |
7.5 |
Temperature(°C) |
25.6 |
25.4 |
25.6 |
25.5 |
25.5 |
|
Dissolved oxygen (%) |
76 |
77 |
76 |
77 |
77 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
|
Day 14 (Spent) |
pH |
7 |
7.2 |
7.1 |
7.1 |
7.1 |
Temperature(°C) |
25.3 |
25.3 |
25.4 |
25.4 |
25.3 |
|
Dissolved oxygen (%) |
72 |
72 |
70 |
71 |
70 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 14 (Fresh) |
pH |
7.6 |
7.6 |
7.6 |
7.7 |
7.7 |
Temperature(°C) |
25.2 |
25.3 |
25.2 |
25.2 |
25.3 |
|
Dissolved oxygen (%) |
76 |
77 |
76 |
76 |
76 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 16 (Spent) |
pH |
7.1 |
7.1 |
7.2 |
7.2 |
7.2 |
Temperature(°C) |
25.4 |
25.6 |
25.5 |
25.5 |
25.6 |
|
Dissolved oxygen (%) |
68 |
69 |
68 |
69 |
69 |
|
Hardness(CaCO3) |
180 |
180 |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
(Continued) The Physico-Chemical Parameters – Range Finding Test
Day |
Parameter |
Concentrations (mg/L) |
||||
Control |
Solvent control |
0.01 |
0.10 |
1.00 |
||
Day 16 (Fresh) |
pH |
7.6 |
7.7 |
7.7 |
7.6 |
7.6 |
Temperature(°C) |
24.9 |
24.8 |
24.9 |
24.8 |
24.8 |
|
Dissolved oxygen (%) |
77 |
76 |
76 |
77 |
76 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 18 (Spent) |
pH |
7.3 |
7.2 |
7.2 |
7.3 |
7.2 |
Temperature(°C) |
25.2 |
25.3 |
25.2 |
25.2 |
25.2 |
|
Dissolved oxygen (%) |
69 |
69 |
69 |
69 |
69 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 18 (Fresh) |
pH |
7.8 |
7.8 |
7.8 |
7.8 |
7.8 |
Temperature(°C) |
25.7 |
25.6 |
25.7 |
25.7 |
25.6 |
|
Dissolved oxygen (%) |
77 |
76 |
76 |
76 |
77 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 20 (Spent) |
pH |
7.3 |
7.3 |
7.3 |
7.3 |
7.2 |
Temperature(°C) |
25.6 |
25.8 |
25.8 |
25.8 |
25.7 |
|
Dissolved oxygen (%) |
70 |
70 |
71 |
71 |
71 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 22 (Fresh) |
pH |
7.7 |
7.7 |
7.6 |
7.7 |
7.7 |
Temperature(°C) |
25.3 |
25.3 |
25.4 |
25.5 |
25.5 |
|
Dissolved oxygen (%) |
77 |
78 |
78 |
77 |
77 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
|
Day 24 (Spent) |
pH |
7.3 |
7.4 |
7.4 |
7.4 |
7.4 |
Temperature(°C) |
25.1 |
25.1 |
25 |
25 |
25 |
|
Dissolved oxygen (%) |
72 |
73 |
72 |
72 |
72 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 24 (Fresh) |
pH |
7.6 |
7.7 |
7.7 |
7.8 |
7.8 |
Temperature(°C) |
25.8 |
25.9 |
25.9 |
25.8 |
25.8 |
|
Dissolved oxygen (%) |
77 |
78 |
77 |
77 |
77 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 26 (Spent) |
pH |
7.2 |
7.2 |
7.2 |
7.3 |
7.2 |
Temperature(°C) |
25.9 |
26 |
26.1 |
26 |
25.9 |
|
Dissolved oxygen (%) |
70 |
70 |
70 |
70 |
70 |
|
Hardness(CaCO3) |
180 |
180 |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
(Continued) The Physico-Chemical Parameters – Range Finding Test
Day |
Parameter |
Concentrations (mg/L) |
||||
Control |
Solvent control |
0.01 |
0.10 |
1.00 |
||
Day 26 (Fresh) |
pH |
7.6 |
7.6 |
7.7 |
7.7 |
7.6 |
Temperature(°C) |
25.7 |
25.6 |
25.7 |
25.6 |
25.7 |
|
Dissolved oxygen (%) |
77 |
75 |
75 |
75 |
75 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 28 (Spent) |
pH |
7.1 |
7.2 |
7.1 |
7.1 |
7.2 |
Temperature(°C) |
25.6 |
25.8 |
25.8 |
25.7 |
25.7 |
|
Dissolved oxygen (%) |
68 |
69 |
69 |
69 |
68 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 28 (Fresh) |
pH |
7.6 |
7.7 |
7.7 |
7.7 |
7.7 |
Temperature(°C) |
25.1 |
24.9 |
24.9 |
24.9 |
25 |
|
Dissolved oxygen (%) |
78 |
78 |
77 |
78 |
77 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
|
Day 30 (Spent) |
pH |
7.1 |
7.2 |
7.2 |
7.1 |
7.1 |
Temperature(°C) |
25.2 |
25.3 |
25.3 |
25.2 |
25.3 |
|
Dissolved oxygen (%) |
70 |
71 |
71 |
71 |
71 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Day 30 (Fresh) |
pH |
7.8 |
7.6 |
7.6 |
7.7 |
7.7 |
Temperature(°C) |
25.7 |
25.8 |
25.7 |
25.8 |
25.6 |
|
Dissolved oxygen (%) |
77 |
78 |
78 |
77 |
77 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 32 (Spent) |
pH |
7.3 |
7.4 |
7.3 |
7.3 |
7.3 |
Temperature(°C) |
25.2 |
25 |
25 |
25.1 |
25.1 |
|
Dissolved oxygen (%) |
72 |
72 |
72 |
73 |
73 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
|
Day 32 (Fresh) |
pH |
7.7 |
7.8 |
7.7 |
7.7 |
7.8 |
Temperature(°C) |
24.9 |
24.8 |
24.9 |
24.8 |
24.8 |
|
Dissolved oxygen (%) |
78 |
78 |
77 |
77 |
77 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
|
Day 33 (Spent) |
pH |
7.3 |
7.2 |
7.2 |
7.3 |
7.2 |
Temperature(°C) |
25.1 |
25.2 |
25.1 |
25.2 |
25.2 |
|
Dissolved oxygen (%) |
72 |
71 |
71 |
72 |
72 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
10 The Physico-Chemical Parameters – Definitive Test
Hours/Day |
Parameter |
Concentrations (mg/L) |
||||||
Control |
Solvent control |
0.01 |
0.03 |
0.10 |
0.30 |
1.00 |
||
0 hr (Day 0) (Fresh) |
pH |
7.8 |
7.8 |
7.8 |
7.8 |
7.7 |
7.8 |
7.8 |
Temperature(°C) |
25.8 |
25.8 |
25.8 |
25.8 |
25.7 |
25.7 |
25.6 |
|
Dissolved oxygen (%) |
78 |
78 |
78 |
78 |
78 |
78 |
76 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
48 hr (Day 2) (Spent) |
pH |
7.6 |
7.6 |
7.5 |
7.5 |
7.5 |
7.5 |
7.5 |
Temperature(°C) |
25.1 |
25.1 |
25.0 |
25.0 |
25.0 |
25.0 |
25.0 |
|
Dissolved oxygen (%) |
74 |
74 |
73 |
73 |
72 |
73 |
73 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
48 hr (Day 2) (Fresh) |
pH |
7.7 |
7.7 |
7.7 |
7.7 |
7.8 |
7.7 |
7.8 |
Temperature(°C) |
26.0 |
26.0 |
25.9 |
25.9 |
25.9 |
25.9 |
25.9 |
|
Dissolved oxygen (%) |
76 |
76 |
76 |
76 |
76 |
76 |
76 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
96 hr (Day 4) Spent |
pH |
7.5 |
7.4 |
7.4 |
7.5 |
7.4 |
7.5 |
7.5 |
Temperature(°C) |
25.7 |
25.7 |
25.6 |
25.6 |
25.5 |
25.6 |
25.6 |
|
Dissolved oxygen (%) |
71 |
71 |
71 |
70 |
70 |
70 |
70 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
96 hr (Day 4) Fresh |
pH |
7.7 |
7.8 |
7.7 |
7.7 |
7.8 |
7.8 |
7.8 |
Temperature(°C) |
25.4 |
25.4 |
25..4 |
25.4 |
25.3 |
25.3 |
25.3 |
|
Dissolved oxygen (%) |
75 |
75 |
75 |
74 |
74 |
74 |
74 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 6 (Spent) |
pH |
7.6 |
7.6 |
7.5 |
7.5 |
7.4 |
7.4 |
7.4 |
Temperature(°C) |
25.2 |
25.2 |
25.3 |
25.2 |
25.2 |
25.2 |
25.2 |
|
Dissolved oxygen (%) |
69 |
69 |
69 |
68 |
68 |
68 |
68 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
- |
- |
|
Day 6 (Fresh) |
pH |
7.7 |
7.7 |
7.7 |
7.6 |
7.7 |
7.7 |
7.7 |
Temperature(°C) |
25.7 |
25.7 |
25.8 |
25.8 |
25.7 |
25.7 |
25.7 |
|
Dissolved oxygen (%) |
74 |
74 |
74 |
74 |
74 |
74 |
74 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 8 (Spent) |
pH |
7.5 |
7.6 |
7.5 |
7.5 |
7.5 |
7.5 |
7.5 |
Temperature(°C) |
25.4 |
25.4 |
25.4 |
25.4 |
25.3 |
25.3 |
25.3 |
|
Dissolved oxygen (%) |
68 |
68 |
68 |
68 |
68 |
68 |
68 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
(Continued) The Physico-Chemical Parameters – Definitive Test
Day |
Parameter |
Concentrations (mg/L) |
||||||
Control |
Solvent control |
0.01 |
0.03 |
0.10 |
0.30 |
1.00 |
||
Day 8 (Fresh) |
pH |
7.7 |
7.6 |
7.6 |
7.7 |
7.6 |
7.6 |
7.6 |
Temperature(°C) |
25.8 |
25.8 |
25.8 |
25.8 |
25.7 |
25.7 |
25.7 |
|
Dissolved oxygen (%) |
77 |
77 |
78 |
77 |
77 |
77 |
77 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
Day 10 (Spent) |
pH |
7.4 |
7.5 |
7.5 |
7.4 |
7.4 |
7.4 |
7.3 |
Temperature(°C) |
25.2 |
25.2 |
25.2 |
25.2 |
25.1 |
25.1 |
25.1 |
|
Dissolved oxygen (%) |
73 |
73 |
74 |
73 |
73 |
73 |
73 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 10 (Fresh) |
pH |
7.6 |
7.6 |
7.6 |
7.7 |
7.7 |
7.8 |
7.7 |
Temperature(°C) |
26.0 |
26.0 |
26.0 |
26.0 |
26.0 |
26.1 |
26.1 |
|
Dissolved oxygen (%) |
77 |
77 |
77 |
77 |
77 |
77 |
77 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
Day 12 (Spent) |
pH |
7.4 |
7.4 |
7.3 |
7.4 |
7.4 |
7.4 |
7.3 |
Temperature(°C) |
25.5 |
25.5 |
25.5 |
25.6 |
25.6 |
25.6 |
25.5 |
|
Dissolved oxygen (%) |
73 |
72 |
73 |
72 |
72 |
72 |
71 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Day 12 (Fresh) |
pH |
7.6 |
7.7 |
7.7 |
7.6 |
7.6 |
7.6 |
7.7 |
Temperature(°C) |
25.5 |
25.5 |
25.5 |
25.5 |
25.5 |
25.5 |
25.4 |
|
Dissolved oxygen (%) |
77 |
76 |
76 |
75 |
75 |
75 |
74 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 14 (Spent) |
pH |
7.3 |
7.3 |
7.3 |
7.3 |
7.3 |
7.3 |
7.2 |
Temperature(°C) |
25.2 |
25.2 |
25.3 |
25.3 |
25.3 |
25.3 |
25.2 |
|
Dissolved oxygen (%) |
73 |
73 |
72 |
72 |
72 |
72 |
72 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Day 14 (Fresh) |
pH |
7.7 |
7.7 |
7.6 |
7.6 |
7.6 |
7.6 |
7.6 |
Temperature(°C) |
25.5 |
25.5 |
25.5 |
25.5 |
25.5 |
25.5 |
25.4 |
|
Dissolved oxygen (%) |
77 |
77 |
78 |
77 |
77 |
77 |
77 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
Day 16 (Spent) |
pH |
7.4 |
7.4 |
7.4 |
7.5 |
7.5 |
7.5 |
7.5 |
Temperature(°C) |
25.2 |
25.3 |
25.2 |
25.3 |
25.3 |
25.3 |
25.3 |
|
Dissolved oxygen (%) |
69 |
68 |
68 |
68 |
68 |
68 |
68 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
(Continued) The Physico-Chemical Parameters – Definitive Test
Day |
Parameter |
Concentrations (mg/L) |
||||||
Control |
Solvent control |
0.01 |
0.03 |
0.10 |
0.30 |
1.00 |
||
Day 16 (Fresh) |
pH |
7.8 |
7.7 |
7.7 |
7.7 |
7.7 |
7.7 |
7.7 |
Temperature(°C) |
25.7 |
25.7 |
25.7 |
25.7 |
25.7 |
25.7 |
25.7 |
|
Dissolved oxygen (%) |
76 |
76 |
76 |
76 |
76 |
76 |
76 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
Day 18 (Spent) |
pH |
7.4 |
7.4 |
7.4 |
7.3 |
7.3 |
7.3 |
7.3 |
Temperature(°C) |
25.4 |
25.4 |
25.4 |
25.3 |
25.3 |
25.3 |
25.3 |
|
Dissolved oxygen (%) |
68 |
69 |
69 |
69 |
69 |
69 |
69 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 18 (Fresh) |
pH |
7.6 |
7.7 |
7.6 |
7.7 |
7.7 |
7.7 |
7.7 |
Temperature(°C) |
25.6 |
25.5 |
25.6 |
25.7 |
25.7 |
25.7 |
25.7 |
|
Dissolved oxygen (%) |
77 |
78 |
78 |
77 |
77 |
77 |
77 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 20 (Spent) |
pH |
7.2 |
7.2 |
7.3 |
7.3 |
7.3 |
7.3 |
7.3 |
Temperature(°C) |
25.3 |
25.3 |
25.2 |
25.2 |
25.2 |
25.2 |
25.2 |
|
Dissolved oxygen (%) |
71 |
71 |
71 |
69 |
69 |
69 |
69 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
- |
- |
|
Day 22 (Fresh) |
pH |
7.6 |
7.7 |
7.7 |
7.6 |
7.6 |
7.6 |
7.6 |
Temperature(°C) |
25.7 |
25.6 |
25.5 |
25.5 |
25.5 |
25.5 |
25.5 |
|
Dissolved oxygen (%) |
78 |
78 |
78 |
78 |
78 |
78 |
78 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 24 (Spent) |
pH |
7.6 |
7.6 |
7.5 |
7.5 |
7.5 |
7.5 |
7.5 |
Temperature(°C) |
25.1 |
25.1 |
25.0 |
25.0 |
25.0 |
25.0 |
25.0 |
|
Dissolved oxygen (%) |
74 |
74 |
73 |
73 |
73 |
73 |
73 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Day 24 (Fresh) |
pH |
7.8 |
7.7 |
7.7 |
7.7 |
7.7 |
7.7 |
7.7 |
Temperature(°C) |
26.0 |
26.0 |
26.0 |
26.0 |
26.0 |
26.0 |
26.0 |
|
Dissolved oxygen (%) |
78 |
78 |
78 |
77 |
77 |
77 |
77 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 26 (Spent) |
pH |
7.4 |
7.4 |
7.4 |
7.3 |
7.3 |
7.3 |
7.3 |
Temperature(°C) |
25.7 |
25.6 |
25.5 |
25.6 |
25.6 |
25.6 |
25.6 |
|
Dissolved oxygen (%) |
71 |
71 |
70 |
69 |
69 |
69 |
69 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
(Continued) The Physico-Chemical Parameters – Definitive Test
Day |
Parameter |
Concentrations (mg/L) |
||||||
Control |
Solvent control |
0.01 |
0.03 |
0.10 |
0.30 |
1.00 |
||
Day 26 (Fresh) |
pH |
7.7 |
7.7 |
7.7 |
7.6 |
7.6 |
7.6 |
7.6 |
Temperature(°C) |
25.9 |
25.9 |
25.9 |
25.9 |
25.9 |
25.9 |
25.9 |
|
Dissolved oxygen (%) |
76 |
76 |
76 |
75 |
75 |
75 |
75 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
Day 28 (Spent) |
pH |
7.3 |
7.2 |
7.3 |
7.3 |
7.3 |
7.3 |
7.3 |
Temperature(°C) |
25.3 |
25.3 |
25.3 |
25.4 |
25.4 |
25.4 |
25.4 |
|
Dissolved oxygen (%) |
72 |
72 |
71 |
71 |
71 |
71 |
71 |
|
Hardness(CaCO3) |
200 |
210 |
- |
- |
- |
- |
- |
|
Day 28 (Fresh) |
pH |
7.7 |
7.7 |
7.6 |
7.6 |
7.6 |
7.6 |
7.6 |
Temperature(°C) |
25.2 |
25.2 |
25.2 |
25.2 |
25.2 |
25.2 |
25.2 |
|
Dissolved oxygen (%) |
77 |
77 |
77 |
76 |
76 |
76 |
76 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 30 (Spent) |
pH |
7.2 |
7.2 |
7.2 |
7.1 |
7.1 |
7.1 |
7.1 |
Temperature(°C) |
24.7 |
24.7 |
24.7 |
24.8 |
24.8 |
24.8 |
24.8 |
|
Dissolved oxygen (%) |
73 |
72 |
72 |
72 |
72 |
72 |
72 |
|
Hardness(CaCO3) |
190 |
190 |
- |
- |
- |
- |
- |
|
Day 30 (Fresh) |
pH |
7.7 |
7.7 |
7.7 |
7.6 |
7.6 |
7.6 |
7.6 |
Temperature(°C) |
25.9 |
25.9 |
25.9 |
25.9 |
25.9 |
25.9 |
25.9 |
|
Dissolved oxygen (%) |
76 |
76 |
76 |
75 |
75 |
75 |
75 |
|
Hardness(CaCO3) |
220 |
220 |
- |
- |
- |
- |
- |
|
Day 32 (Spen |
pH |
7.2 |
7.2 |
7.2 |
7.1 |
7.1 |
7.1 |
7.1 |
Temperature(°C) |
24.7 |
24.7 |
24.7 |
24.8 |
24.8 |
24.8 |
24.8 |
|
Dissolved oxygen (%) |
73 |
72 |
72 |
72 |
72 |
72 |
72 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Day 32 (Fresh) |
pH |
7.7 |
7.7 |
7.6 |
7.6 |
7.6 |
7.6 |
7.6 |
Temperature(°C) |
25.3 |
25.3 |
25.3 |
25.3 |
25.3 |
25.3 |
25.3 |
|
Dissolved oxygen (%) |
77 |
77 |
77 |
76 |
76 |
76 |
76 |
|
Hardness(CaCO3) |
210 |
210 |
- |
- |
- |
- |
- |
|
Day 33 (Spent |
pH |
7.3 |
7.3 |
7.3 |
7.2 |
7.2 |
7.2 |
7.1 |
Temperature(°C) |
24.8 |
24.8 |
24.8 |
24.8 |
24.8 |
24.7 |
24.7 |
|
Dissolved oxygen (%) |
72 |
72 |
72 |
71 |
71 |
71 |
70 |
|
Hardness(CaCO3) |
200 |
200 |
- |
- |
- |
- |
- |
|
Key: mg- milligram, L- litre, °C – Degree Celsius
10 Pre-hatch observation of embryo –Range Finding Test
Concentration (mg/L) |
R |
No. of embryos exposed |
Pre-hatch |
No. of Dead embryos |
No. of Hatched larvae |
Mean % of Hatched larvae |
||
24h (Day 1) |
48h (Day 2) |
72h (Day 3) |
||||||
Control |
1 |
20 |
20N |
20N |
20N |
0 |
20 |
98.75 |
2 |
20 |
1CE, 19N |
19N |
19N |
1 |
19 |
||
3 |
20 |
20N |
20N |
20N |
0 |
20 |
||
4 |
20 |
20N |
20N |
20N |
0 |
20 |
||
Solvent control |
1 |
20 |
1CE, 19N |
19N |
19N |
1 |
19 |
97.50 |
2 |
20 |
20N |
20N |
20N |
0 |
20 |
||
3 |
20 |
1LS, 19N |
1LS, 19N |
19N |
1 |
19 |
||
4 |
20 |
20N |
20N |
20N |
0 |
20 |
||
0.01 |
1 |
20 |
1CE, 19N |
19N |
19N |
1 |
19 |
95 |
2 |
20 |
1LS, 19N |
1LS, 19N |
11D, 19N |
1 |
19 |
||
3 |
20 |
1LS, 19N |
1D, 19N |
19N |
1 |
19 |
||
4 |
20 |
1CE, 19N |
19N |
19N |
1 |
19 |
||
0.10 |
1 |
20 |
3CE, 2LS, 15N |
2D, 15N |
15N |
5 |
15 |
66.25 |
2 |
20 |
3CE, 2LS, 15N |
1D, 1LS, 15N |
1D, 15N |
5 |
15 |
||
3 |
20 |
5CE, 2LS, 13N |
2D, 13N |
13N |
7 |
13 |
||
4 |
20 |
4CE, 6LS, 10N |
4D, 2LS, 10N |
2D, 10N |
10 |
10 |
||
1.00 |
1 |
20 |
11CE, 2LS, 7N |
2D, 7N |
7N |
13 |
7 |
27.5 |
2 |
20 |
13CE, 1LS, 6N |
1D, 1LH, 5N |
1D, 5N |
15 |
5 |
||
3 |
20 |
12CE, 2LS, 6N |
2D, 2LH, 4N |
2D, 4N |
16 |
4 |
||
4 |
20 |
10CE, 1LS, 9N |
4LH, 6N |
4D, 6N |
14 |
6 |
||
10.00 |
1 |
20 |
20CE |
- |
- |
20 |
0 |
0 |
2 |
20 |
20CE |
- |
- |
20 |
0 |
||
3 |
20 |
20CE |
- |
- |
20 |
0 |
||
4 |
20 |
20CE |
- |
- |
20 |
0 |
||
20.00 |
1 |
20 |
20CE |
- |
- |
20 |
0 |
0 |
2 |
20 |
20CE |
- |
- |
20 |
0 |
||
3 |
20 |
20CE |
- |
- |
20 |
0 |
||
4 |
20 |
20CE |
- |
- |
20 |
0 |
||
Key:mg- milligram; L- litre;CE- Coagulated embryos; LS -
Lack of somite formation; LH - Lack of heartbeat; D – Dead; N –
Normal; R - Replicate
11 Post-hatch Observation of larvae and juveniles–Range Finding Test
Concentration (mg/L) |
R |
No. of hatched Larvae |
Post-Hatch |
||||||||||||||||||||||
Clinical Signs/Dead |
|||||||||||||||||||||||||
Days |
|||||||||||||||||||||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
||||||||||||||||
Control |
1 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
|||||||||||||
2 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
||||||||||||||
3 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
||||||||||||||
4 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
||||||||||||||
Solvent control |
1 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
|||||||||||||
2 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
19N, 1D |
||||||||||||||
3 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
||||||||||||||
4 |
20 |
20N |
20N |
20N |
20N |
20N |
19N, 1D |
19N |
19N |
19N |
19N |
||||||||||||||
0.01 |
1 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
|||||||||||||
2 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
||||||||||||||
3 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
18N |
||||||||||||||
4 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
18N |
||||||||||||||
0.10 |
1 |
15 |
15N |
15N |
15N |
14N, 1D |
14N |
14N |
14N |
14N |
14N |
14N |
|||||||||||||
2 |
15 |
15N |
15N |
14N, 1D |
14N |
14N |
14N |
14N |
14N |
13N, 1D |
13N |
||||||||||||||
3 |
13 |
13N |
13N |
12N, 1D |
12N |
12N |
12N |
12N |
12N |
12N |
11N, 1D |
||||||||||||||
4 |
10 |
10N |
10N |
10N |
10N |
10N |
10N |
9N, 1D |
9N |
9N |
9N |
||||||||||||||
1.00 |
1 |
7 |
7N |
7N |
7N |
7N |
7N |
6N, 1D |
6N |
6N |
6N |
5N, 1D |
|||||||||||||
2 |
5 |
5N |
5N |
5N |
5N |
5N |
5N |
5N |
5N |
4N, 1D |
4N |
||||||||||||||
3 |
4 |
4N |
4N |
4N |
4N |
4N |
4N |
4N |
4N |
4N |
3N, 1D |
||||||||||||||
4 |
6 |
6N |
6N |
6N |
6N |
6N |
6N |
5N, 1D |
5N |
5N |
5N |
||||||||||||||
Key:mg- milligram;N – Normal; D – Dead; R – Replicate
(Continued) Post-hatch Observation of larvae and juveniles–Range Finding Test
Concentration (mg/L) |
R |
No. of hatched larvae |
Post-Hatch |
||||||||||||||||||||||
Clinical Signs/Dead |
|||||||||||||||||||||||||
Days |
|||||||||||||||||||||||||
11 |
12 |
13 |
14 |
15 |
16 |
17 |
18 |
19 |
20 |
||||||||||||||||
Control |
1 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
|||||||||||||
2 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
||||||||||||||
3 |
20 |
20N |
20N |
19N, 1D |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
||||||||||||||
4 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
19N, 1D |
19N |
19N |
||||||||||||||
Solvent control |
1 |
19 |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
18N |
18N |
18N |
18N |
|||||||||||||
2 |
20 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
||||||||||||||
3 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
||||||||||||||
4 |
20 |
19N |
19N |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
18N |
18N |
||||||||||||||
0.01 |
1 |
19 |
18N |
18N |
18N |
18N |
18N |
17N, 1D |
17N |
17N |
17N |
17N |
|||||||||||||
2 |
19 |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
18N |
18N |
18N |
18N |
||||||||||||||
3 |
19 |
18N |
18N |
18N |
17N, 1D |
17N |
17N |
17N |
16N, 1D |
16N |
16N |
||||||||||||||
4 |
19 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
17N, 1D |
17N |
||||||||||||||
0.10 |
1 |
15 |
14N |
13N, 1D |
13N |
13N |
13N |
12N, 1D |
12N |
12N |
12N |
12N |
|||||||||||||
2 |
15 |
13N |
13N |
12N, 1D |
12N |
12N |
12N |
11N, 1D |
11N |
10N, 1D |
10N |
||||||||||||||
3 |
13 |
11N |
11N |
10N, 1D |
10N |
10N |
10N |
10N |
10N |
9N, 1D |
9N |
||||||||||||||
4 |
10 |
9N |
9N |
9N |
9N |
8N, 1D |
8N |
8N |
8N |
8N |
8N |
||||||||||||||
1.00 |
1 |
7 |
5N |
5N |
5N |
5N |
5N |
5N |
5N |
4N, 1D |
4N |
4N |
|||||||||||||
2 |
5 |
4N |
4N |
3N, 1D |
3N |
3N |
3N |
3N |
3N |
3N |
2N, 1D |
||||||||||||||
3 |
4 |
3N |
3N |
3N |
3N |
3N |
3N |
3N |
3N |
2N, 1D |
2N |
||||||||||||||
4 |
6 |
5N |
5N |
5N |
4N, 1D |
4N |
4N |
4N |
4N |
4N |
4N |
||||||||||||||
Key:mg- milligram;N – Normal; D – Dead; R – Replicate
(Continued) Post-hatch Observation of larvae and juveniles–Range Finding Test
Concentration (mg/L) |
R |
No. of hatched larvae |
Post-Hatch |
No. of juveniles surviving at 30 days post-hatch |
Mean % of fish larvae surviving at 30 days post-hatch |
||||||||||||||||||||||||
Clinical Signs/Dead |
|||||||||||||||||||||||||||||
Days |
|||||||||||||||||||||||||||||
21 |
22 |
23 |
24 |
25 |
26 |
27 |
28 |
29 |
30 |
||||||||||||||||||||
Control |
1 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20 |
96.20 |
|||||||||||||||
2 |
19 |
18N, 1D |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
|||||||||||||||||
3 |
20 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19 |
|||||||||||||||||
4 |
20 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19 |
|||||||||||||||||
Solvent control |
1 |
19 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
93.59 |
|||||||||||||||
2 |
20 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19 |
|||||||||||||||||
3 |
19 |
19N |
18N, 1D |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
|||||||||||||||||
4 |
20 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
|||||||||||||||||
0.01 |
1 |
19 |
17N |
17N |
16N, 1D |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
16 |
84.21 |
|||||||||||||||
2 |
19 |
17N, 1D |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17 |
|||||||||||||||||
3 |
19 |
16N |
16N |
15N, 1D |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15 |
|||||||||||||||||
4 |
19 |
17N |
17N |
17N |
16N, 1D |
16N |
16N |
16N |
16N |
16N |
16N |
16 |
|||||||||||||||||
0.10 |
1 |
15 |
11N, 1D |
11N |
11N |
10N, 1D |
10N |
10N |
10N |
10N |
10N |
10N |
10 |
64.15 |
|||||||||||||||
2 |
15 |
10N |
9N, 1D |
9N |
9N |
9N |
9N |
9N |
9N |
9N |
9N |
9 |
|||||||||||||||||
3 |
13 |
9N |
9N |
8N, 1D |
8N |
8N |
8N |
8N |
8N |
8N |
8N |
8 |
|||||||||||||||||
4 |
10 |
8N |
7N, 1D |
7N |
7N |
7N |
7N |
7N |
7N |
7N |
7N |
7 |
|||||||||||||||||
1.00 |
1 |
7 |
3N , 1D |
3N |
2N, 1D |
2N |
2N |
2N |
2N |
2N |
2N |
2N |
2 |
31.82 |
|||||||||||||||
2 |
5 |
2N |
2N |
2N |
2N |
1N, 1D |
1N |
1N |
1N |
1N |
1N |
1 |
|||||||||||||||||
3 |
4 |
2N |
2N |
2N |
1N, 1D |
1N |
1N |
1N |
1N |
1N |
1N |
1 |
|||||||||||||||||
4 |
6 |
4N |
4N |
4N |
4N |
3N, 1D |
3N |
3N |
3N |
3N |
3N |
3 |
|||||||||||||||||
Key:mg- milligram;N – Normal; H-Hypoventilation; L – Lethargy; D – Dead; R – Replicate
12 Stage wise mortality of fish- Range Finding Test
Concentration (mg/L) |
R |
No. of eggs added at the start of test |
Mortality |
||||||||||||||||||||||||||||||||||||||
Pre-hatch |
Post-hatch |
||||||||||||||||||||||||||||||||||||||||
Embryo stage |
Larva and Juveniles stage |
||||||||||||||||||||||||||||||||||||||||
Days |
|||||||||||||||||||||||||||||||||||||||||
1 |
2 |
3 |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
13 |
14 |
15 |
||||||||||||||||||||||||
Control |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
|||||||||||||||||||||
2 |
20 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
||||||||||||||||||||||
3 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
||||||||||||||||||||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
||||||||||||||||||||||
Solvent control |
1 |
20 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
|||||||||||||||||||||
2 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
||||||||||||||||||||||
3 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
||||||||||||||||||||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
||||||||||||||||||||||
0.01 |
1 |
20 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
|||||||||||||||||||||
2 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
||||||||||||||||||||||
3 |
20 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
||||||||||||||||||||||
4 |
20 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
||||||||||||||||||||||
0.10 |
1 |
20 |
3 |
2 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
|||||||||||||||||||||
2 |
20 |
3 |
1 |
1 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
1 |
0 |
0 |
||||||||||||||||||||||
3 |
20 |
5 |
2 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
1 |
0 |
0 |
||||||||||||||||||||||
4 |
20 |
4 |
4 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
||||||||||||||||||||||
1.00 |
1 |
20 |
11 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
|||||||||||||||||||||
2 |
20 |
13 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
1 |
0 |
0 |
||||||||||||||||||||||
3 |
20 |
12 |
2 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
||||||||||||||||||||||
4 |
20 |
10 |
0 |
4 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
||||||||||||||||||||||
10.00 |
1 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
|||||||||||||||||||||
2 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||||||||
3 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||||||||
4 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||||||||
20.00 |
1 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
|||||||||||||||||||||
2 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||||||||
3 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||||||||
4 |
20 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||||||||
Key:mg- milligram;R – Replicate
(Continued) Stage wise mortality of fish - Range Finding Test
Concentration (mg/L) |
R |
No. of eggs added at the start of test |
Mortality |
Cumulative Mortality (Day 30) |
Total No. of cumulative mortality |
Mean % of Cumulative Mortality |
SD |
||||||||||||||||||||||||||||||||||||
Larva and Juveniles stage |
|||||||||||||||||||||||||||||||||||||||||||
Days |
|||||||||||||||||||||||||||||||||||||||||||
16 |
17 |
18 |
19 |
20 |
21 |
22 |
23 |
24 |
25 |
26 |
27 |
28 |
29 |
30 |
|
|
|
|
|||||||||||||||||||||||||
Control |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
5 |
6.25 |
0.5 |
||||||||||||||||||||||
2 |
20 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
||||||||||||||||||||||||||
3 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
||||||||||||||||||||||||||
4 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
||||||||||||||||||||||||||
Solvent control |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
7 |
8.75 |
0.5 |
||||||||||||||||||||||
2 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
||||||||||||||||||||||||||
3 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
||||||||||||||||||||||||||
4 |
20 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
||||||||||||||||||||||||||
0.01 |
1 |
20 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
4 |
16 |
22.5 |
0.8 |
||||||||||||||||||||||
2 |
20 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
3 |
||||||||||||||||||||||||||
3 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
5 |
||||||||||||||||||||||||||
4 |
20 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
4 |
||||||||||||||||||||||||||
0.10 |
1 |
20 |
1 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
10 |
46 |
57.5 |
1.3 |
||||||||||||||||||||||
2 |
20 |
0 |
1 |
0 |
1 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
11 |
||||||||||||||||||||||||||
3 |
20 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
12 |
||||||||||||||||||||||||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
13 |
||||||||||||||||||||||||||
1.00 |
1 |
20 |
0 |
0 |
1 |
0 |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
18 |
72 |
90 |
0.8 |
||||||||||||||||||||||
2 |
20 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
18 |
||||||||||||||||||||||||||
3 |
20 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
19 |
||||||||||||||||||||||||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
17 |
||||||||||||||||||||||||||
10.00 |
1 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
80 |
100 |
0 |
||||||||||||||||||||||
2 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
||||||||||||||||||||||||||
3 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
||||||||||||||||||||||||||
4 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
||||||||||||||||||||||||||
20.00 |
1 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
80 |
100 |
0 |
||||||||||||||||||||||
2 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
||||||||||||||||||||||||||
3 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
||||||||||||||||||||||||||
4 |
20 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
20 |
||||||||||||||||||||||||||
Key:mg- milligram;R – Replicate; SD – Standard Deviation
13 Pre-hatch observation of embryo – Definitive Test
Concentration (mg/L) |
R |
No. of embryos exposed |
Pre-hatch |
No. of Dead embryos |
No. of Hatched larvae |
Mean % of Hatched larvae |
|||||||||||
24h (Day 1) |
48h (Day 2) |
72h (Day 3) |
|||||||||||||||
Control |
1 |
20 |
20N |
20N |
20N |
0 |
20 |
95.00 |
|||||||||
2 |
20 |
20N |
1LH, 19N |
1LH, 19N |
1 |
19 |
|||||||||||
3 |
20 |
20N |
1LS, 19N |
1LS, 19N |
1 |
19 |
|||||||||||
4 |
20 |
1LS, 19N |
1LS, 1LH, 18N |
1LS, 1LH, 18N |
2 |
18 |
|||||||||||
Solvent control |
1 |
20 |
1LS, 19N |
1LS, 19N |
1LS, 19N |
1 |
19 |
93.75 |
|||||||||
2 |
20 |
20N |
20N |
20N |
0 |
20 |
|||||||||||
3 |
20 |
20N |
1LS, 1LH, 18N |
1LS, 1LH, 18N |
2 |
18 |
|||||||||||
4 |
20 |
1LS, 19N |
1LS, 1LH, 18N |
1LS, 1LH, 18N |
2 |
18 |
|||||||||||
0.01 |
1 |
20 |
1CE, 2LS, 17N |
2LS, 17N |
2LS, 17N |
3 |
17 |
85.00 |
|||||||||
2 |
20 |
2CE, 1LS, 17N |
1LS, 17N |
1LS, 17N |
3 |
17 |
|||||||||||
3 |
20 |
2LS, 1CE, 16N |
2LS, 1LH, 16N |
2LS, 1LH, 16N |
4 |
16 |
|||||||||||
4 |
20 |
2LS, 18N |
2LS, 18N |
2LS, 18N |
2 |
18 |
|||||||||||
0.03 |
1 |
20 |
4CE, 2LS, 14N |
2LS, 14N |
2LS, 14N |
6 |
14 |
72.50 |
|||||||||
2 |
20 |
4CE, 1LS, 15N |
1LS, 15N |
1LS, 15N |
5 |
15 |
|||||||||||
3 |
20 |
3LS, 3CE, 14N |
3LS, 14N |
3LS, 14N |
6 |
14 |
|||||||||||
4 |
20 |
3CE, 2LS, 15N |
2LS, 15N |
2LS, 15N |
5 |
15 |
|||||||||||
0.10 |
1 |
20 |
6CE, 2LS, 12N |
2LS, 12N |
2LS, 12N |
8 |
12 |
66.25 |
|||||||||
2 |
20 |
5CE, 1LS, 14N |
1LS, 14N |
1LS, 14N |
6 |
14 |
|||||||||||
3 |
20 |
3CE, 2LS, 15N |
1LS, 1LH, 15N |
1LS, 1LH, 15N |
5 |
15 |
|||||||||||
4 |
20 |
5CE, 3LS, 12N |
3LS, 12N |
3LS, 12N |
8 |
12 |
|||||||||||
0.30 |
1 |
20 |
7CE, 2LS, 11N |
2LS, 11N |
2LS, 11N |
9 |
11 |
53.75 |
|||||||||
2 |
20 |
8CE, 2LS, 10N |
2LS, 10N |
2LS, 10N |
10 |
10 |
|||||||||||
3 |
20 |
7CE, 1LS, 12N |
1LS, 12N |
1LS, 12N |
8 |
12 |
|||||||||||
4 |
20 |
8CE, 2LS, 10N |
2LS, 10N |
2LS, 10N |
10 |
10 |
|||||||||||
1.00 |
1 |
20 |
14CE, 2LS, 4N |
2LS, 4N |
2LS, 4N |
16 |
4 |
25.00 |
|||||||||
2 |
20 |
13CE, 2LS, 5N |
2LS, 5N |
2LS, 5N |
15 |
5 |
|||||||||||
3 |
20 |
14CE, 1LS, 5N |
1LS, 5N |
1LS, 5N |
15 |
5 |
|||||||||||
4 |
20 |
12CE, 2LS, 6N |
2LS, 6N |
2LS, 6N |
14 |
6 |
|||||||||||
Key:mg-
milligram; L- litre;CE- Coagulated embryos; LS - Lack of somite
formation; LH - Lack of heartbeat; N – Normal; R – Replicate
14 Post-hatch Observation of larvae and juveniles–Definitive Test
Concentration (mg/L) |
R |
No. of hatched Larvae |
Post-Hatch |
|||||||||
Clinical Signs/Dead |
||||||||||||
Days |
||||||||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
|||
Control |
1 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
2 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
|
3 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
18N |
18N, |
|
4 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
|
Solvent control |
1 |
19 |
19N |
19N |
19N |
19N |
19N |
18N, 1D |
18N |
18N |
18N |
18N |
2 |
20 |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
20N |
|
3 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
|
4 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
|
0.01 |
1 |
17 |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
2 |
17 |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
|
3 |
16 |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
|
4 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
17N, 1D |
17N |
17N |
|
0.03 |
1 |
14 |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
13N, 1D |
2 |
15 |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
|
3 |
14 |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
|
4 |
15 |
15N |
15N |
15N |
15N |
15N |
15N |
14N, 1D |
15N |
15N |
15N |
|
0.10 |
1 |
12 |
12N |
12N |
12N |
12N |
12N |
12N |
11N, 1D |
11N |
11N |
11N |
2 |
14 |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
|
3 |
15 |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
14N, 1D |
14N |
|
4 |
12 |
12N |
12N |
12N |
12N |
12N |
12N |
12N |
12N |
12N |
11N, 1D |
|
0.30 |
1 |
11 |
11N |
11N |
11N |
11N |
10N, 1D |
10N |
10N |
10N |
10N |
9N, 1D |
2 |
10 |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
|
3 |
12 |
12N |
12N |
12N |
11N, 1D |
11N |
11N |
11N |
11N |
11N |
11N |
|
4 |
10 |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
|
1.00 |
1 |
4 |
4N |
4N |
4N |
4N |
4N |
4N |
3N, 1D |
3N |
3N |
3N |
2 |
5 |
5N |
5N |
5N |
5N |
5N |
5N |
5N |
4N, 1D |
4N |
4N |
|
3 |
5 |
5N |
5N |
5N |
5N |
5N |
5N |
5N |
4N, 1D |
4N |
4N |
|
4 |
6 |
6N |
6N |
6N |
6N |
6N |
6N |
6N |
6N |
6N |
6N |
|
Key:mg- milligram;N – Normal; D – Dead; R – Replicate
(Continued) Post-hatch Observation of larvae and juveniles– Definitive Test
Concentration (mg/L) |
R |
No. of hatched larvae |
Post-Hatch |
|||||||||
Clinical Signs/Dead |
||||||||||||
Days |
||||||||||||
11 |
12 |
13 |
14 |
15 |
16 |
17 |
18 |
19 |
20 |
|||
Control |
1 |
20 |
20N |
20N |
20N |
19N, 1D |
19N |
19N |
19N |
19N |
19N |
19N |
2 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
|
3 |
19 |
18N, |
18N, |
18N, |
18N, |
18N |
18N |
17N, 1D |
17N |
17N |
17N |
|
4 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
|
Solvent control |
1 |
19 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
2 |
20 |
20N |
20N |
20N |
20N |
19N, 1D |
19N |
19N |
19N |
18N, 1D |
18N |
|
3 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
|
4 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
|
0.01 |
1 |
17 |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
2 |
17 |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
|
3 |
16 |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
|
4 |
18 |
17N |
17N |
17N |
17N |
16N, 1D |
16N |
16N |
16N |
16N |
16N |
|
0.03 |
1 |
14 |
13N |
13N |
13N |
13N |
13N, |
13N |
13N |
13N |
13N |
13N |
2 |
15 |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
|
3 |
14 |
14N |
14N |
14N |
14N |
14N |
14N |
13N, 1D |
13N |
13N |
13N |
|
4 |
15 |
15N |
15N |
15N |
15N |
15N |
14N, 1D |
14N |
14N |
14N |
14N |
|
0.10 |
1 |
12 |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
2 |
14 |
14N |
14N |
14N |
14N |
14N |
14N |
13N, 1D |
13N |
12N, 1L |
12N, 1D |
|
3 |
15 |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
13N, 1D |
|
4 |
12 |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
|
0.30 |
1 |
11 |
9N |
9N |
9N |
9N |
9N |
9N |
9N |
9N |
9N |
9N |
2 |
10 |
10N |
10N |
10N |
10N |
10N |
10N |
9N, 1D |
9N |
9N |
9N |
|
3 |
12 |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
11N |
10N, 1D |
10N |
|
4 |
10 |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10N |
10 |
9N, 1D |
|
1.00 |
1 |
4 |
3N |
3N |
3N |
3N |
3N |
3N |
3N |
3N |
3N |
3N |
2 |
5 |
4N |
4N |
4N |
4N |
4N |
4N |
4N |
3N, 1D |
3N |
3N |
|
3 |
5 |
4N |
4N |
4N |
4N |
4N |
4N |
3N, 1D |
3N |
3N |
3N |
|
4 |
6 |
6N |
6N |
6N |
6N |
6N |
5N, 1D |
5N |
5N |
4N, 1D |
4N |
|
Key:mg- milligram;N – Normal; D – Dead; R – Replicate
(Continued) Post-hatch Observation of larvae and juveniles– Definitive Test
Concentration (mg/L) |
R |
No. of hatched larvae |
Post-Hatch |
No. of juveniles surviving at 30 days post-hatch |
Mean % of fish larvae surviving at 30 days post-hatch |
|||||||||
Clinical Signs/Dead |
||||||||||||||
Days |
||||||||||||||
21 |
22 |
23 |
24 |
25 |
26 |
27 |
28 |
29 |
30 |
|||||
Control |
1 |
20 |
19N |
19N |
19N |
19N |
19N |
18N, 1L |
18N, 1H |
18N, 1D |
18N |
18N |
18 |
93.42 |
2 |
19 |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19N |
19 |
||
3 |
19 |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
16N, 1L |
16N, 1H |
16N, 1D |
16 |
||
4 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
||
Solvent control |
1 |
19 |
18N |
18N |
18N |
18N |
18N |
17N, 1L |
17N, 1L |
17N, 1H |
17N, 1D |
17N |
17 |
94.67 |
2 |
20 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
||
3 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
||
4 |
18 |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18N |
18 |
||
0.01 |
1 |
17 |
17N |
17N |
17N |
17N |
17N |
16N, 1L |
16N, 1L |
16N, 1D |
16N |
15N, 1L |
16 |
91.18 |
2 |
17 |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17N |
17 |
||
3 |
16 |
16N |
16N |
16N |
16N |
16N |
16N |
16N |
15N, 1L |
15N, 1D |
15N |
15 |
||
4 |
18 |
16N |
16N |
16N |
15N, 1L |
15N, 1D |
14N, 1L |
14N, 1D |
14N |
14N |
13N, 1L |
14 |
||
0.03 |
1 |
14 |
13N |
13N |
13N |
13N |
13N |
13N |
12N, 1L |
12N, 1D |
12N |
12N |
12 |
87.93 |
2 |
15 |
15N |
15N |
15N |
15N |
15N |
15N |
15N |
14N, 1L |
14N, 1D |
14N |
14 |
||
3 |
14 |
13N |
13N |
13N |
13N |
12N, 1L |
12N, 1H |
12N, 1D |
12N |
12N |
12N |
12 |
||
4 |
15 |
14N |
14N |
14N |
14N |
14N |
14N |
14N |
13N, 1L |
13N, 1D |
12N, 1L |
13 |
||
0.10 |
1 |
12 |
11N |
11N |
11N |
10N, 1L |
9N, 2L |
8N, 3L |
8N, 1L, 2D |
8N, 1L |
8N, 1D |
8N |
8 |
67.92 |
2 |
14 |
12N |
13N |
13N |
12N, 1L |
12N, 1D |
12N |
10N, 2L |
10N, 2D |
9N, 1L |
9N, 1D |
9 |
||
3 |
15 |
13N |
13N |
13N |
12N, 1L |
12N, 1H |
12N, 1D |
11N, 1L |
11N, 1D |
11N, 1L |
10N, 1D |
10 |
||
4 |
12 |
11N |
11N |
11N |
11N |
10N, 1D |
10N |
10N |
9N, 1L |
9N, 1D |
8N, 1L |
9 |
||
0.30 |
1 |
11 |
9N |
9N |
9N |
9N |
9N |
8N, 1L |
7N, 2L |
7N, 2L |
7N, 2D |
6N, 1L |
7 |
53.49 |
2 |
10 |
9N |
9N |
9N |
9N |
9N |
9N |
8N, 1L |
8N, 1D |
7N, 1D |
6N, 1D |
6 |
||
3 |
12 |
10N |
10N |
10N |
10N |
9N, 1L |
9N, 1D |
8N, 1L |
8N, 1D |
5N, 3L |
5N, 3D |
5 |
||
4 |
10 |
9N |
9N |
9N |
9N |
9N |
8N, 1L |
7N, 1L, 1D |
7N, 1L |
5N, 3 L |
5N, 3D |
5 |
||
1.00 |
1 |
4 |
3N |
3N |
3N |
3N |
3N |
1N, 2L |
1N, 1L, 1D |
1N, 1L |
1L, 1D |
1L |
1 |
25.00 |
2 |
5 |
3N |
3N |
3N |
3N |
3N |
3N |
2N, 1L |
2N, 1D |
1N, 1L |
2L |
2 |
||
3 |
5 |
3N |
3N |
3N |
3N |
3N |
3N |
1N, 2L |
2L, 1D |
1L, 1D |
1L |
1 |
||
4 |
6 |
4N |
4N |
4N |
4N |
3N, 1L |
2N, 2L |
2N, 1L, 1D |
1N, 2L |
1N, 2L |
1L, 2D |
1 |
||
Key:mg- milligram;N – Normal; H-Hypoventilation; L – Lethargy; D – Dead; R – Replicate
15 Stage wise mortality of fish-Definitive test
Concentration (mg/L) |
R |
No. of eggs added at the start of test |
Mortality |
|||||||||||||||||
Pre-hatch |
Post-hatch |
|||||||||||||||||||
Embryo stage |
Larva and Juveniles stage |
|||||||||||||||||||
Days |
||||||||||||||||||||
1 |
2 |
3 |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
13 |
14 |
15 |
|||
Control |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
2 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
3 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 |
20 |
0 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
Solvent control |
1 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
|
3 |
20 |
0 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 |
20 |
0 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
0.01 |
1 |
20 |
1 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
20 |
2 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
3 |
20 |
1 |
0 |
3 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 |
20 |
0 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
|
0.03 |
1 |
20 |
4 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
2 |
20 |
4 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
3 |
20 |
3 |
0 |
3 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 |
20 |
3 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
0.10 |
1 |
20 |
6 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
20 |
5 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
3 |
20 |
3 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 |
20 |
5 |
0 |
3 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
|
0.30 |
1 |
20 |
7 |
0 |
2 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
2 |
20 |
8 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
3 |
20 |
7 |
0 |
1 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 |
20 |
8 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
1.00 |
1 |
20 |
14 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
20 |
13 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
3 |
20 |
14 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 |
20 |
12 |
0 |
2 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
Key:mg- milligram;R – Replicate
(Continued) Stage wise mortality of fish- Definitive test
Concentration (mg/L) |
R |
No. of eggs added at the start of test |
Mortality |
Cumulative Mortality (Day 30) |
Total No. of cumulative mortality |
Mean % of Cumulative Mortality |
SD |
||||||||||||||
Larva and Juveniles stage |
|||||||||||||||||||||
Days |
|||||||||||||||||||||
16 |
17 |
18 |
19 |
20 |
21 |
22 |
23 |
24 |
25 |
26 |
27 |
28 |
29 |
30 |
|||||||
Control |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
2 |
9 |
11.25 |
1.3 |
2 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
||||
3 |
20 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
4 |
||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
||||
Solvent control |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
3 |
9 |
11.25 |
0.5 |
2 |
20 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
||||
3 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
||||
0.01 |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
4 |
18 |
22.5 |
1.3 |
2 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
3 |
||||
3 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
5 |
||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
6 |
||||
0.03 |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
8 |
29 |
36.5 |
1.0 |
2 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
6 |
||||
3 |
20 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
8 |
||||
4 |
20 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
7 |
||||
0.10 |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
0 |
1 |
0 |
12 |
44 |
55 |
0.8 |
2 |
20 |
0 |
1 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
2 |
0 |
1 |
11 |
||||
3 |
20 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
1 |
10 |
||||
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
1 |
0 |
11 |
||||
0.30 |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
0 |
13 |
57 |
71.25 |
1.0 |
2 |
20 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
1 |
1 |
14 |
||||
3 |
20 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
3 |
15 |
||||
4 |
20 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
3 |
15 |
||||
1.00 |
1 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
19 |
75 |
93.75 |
0.5 |
2 |
20 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
18 |
||||
3 |
20 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
1 |
0 |
19 |
||||
4 |
20 |
1 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
2 |
19 |
||||
Key:mg- milligram;R – Replicate; SD – Standard Deviation
16 Endpoint values for Mortality, Hatching success and Fish larvae survival during the fish early-life stage test– Range finding test
Concentration mg/L |
Mortality |
Hatching success and fish larvae survival |
||||
Total (%) |
Corrected# |
Mean % of hatched larvae |
Fertility reduction (%) |
Mean % of surviving Juveniles |
Surviving juveniles reduction (%) |
|
(%) |
||||||
Control |
6.25 |
- |
98.75 |
- |
96.20 |
- |
Solvent control |
8.75 |
- |
97.50 |
- |
93.59 |
- |
N-(4-((4-hydroxy-9, 10-dioxo-9,10-dihydroanthracen-1-yl) amino) phenyl)acetamide |
||||||
0.01 |
22.50 |
15.07 |
95.00 |
2.56 |
84.21 |
10.18 |
0.10 |
57.50 |
57.50 |
66.25 |
32.05 |
64.15 |
31.57 |
1.00 |
90.00 |
87.10 |
27.50 |
71.79 |
31.82 |
66.06 |
10.00 |
100.00 |
100.00 |
0.00 |
100.00 |
- |
- |
20.00 |
100.00 |
100.00 |
0.00 |
100.00 |
- |
- |
#: Mortality corrected according to Abbott's formula:
Corrected mortality [%] = ((Mt - Mc) ÷ (100 - Mc)) × 100; Mt = Mortality treated, Mc = Mortality Solvent control
Description of key information
The aim of the study was todetermine the EC50, LOEC and NOECof test chemical on the growth and development of embryos and larvae of the freshwater fish species zebrafish (Danio rerio) in a Fish Early-Life Stage (ELS) Toxicity Test. The study was conducted according to the OECD guideline No. 210, Adopted 26thJuly, 2013.
The test was conducted with a semi-static design with renewal once 48 h.
Concentrated stock solutions and test media were analysed during the ELS test. Based on the range finding result the definitive test was conducted with five test concentrations of 0.01, 0.03, 0.10, 0.30 and 1.00 mg/L in terms of test chemical with a conversion factor of 3.2. Both control and solvent control were included in the test design. Each concentration was divided into four replicate with 20 eggs each and including the controls.
The cumulative mortality of the control and solvent control group at 30 days post-hatch was 11.25 and 11.25 %, whereas corrected cumulative mortality of embryos, larvae and juvenile fish after at the end of exposure to test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg /Lwas 12.68, 28.45, 49.30, 67.61and 92.96%,respectively.
Hatching success in the control and solvent control group was 93.42 and 94.67% respectively, satisfying the validation criterion for hatching success (70%),whereas in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas 91.18, 87.93, 67.92, 53.49and 25.00%,respectively. Fertility reduction in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas3.69, 7.12, 28.26, 43.50 and 73.59%,respectively in comparison with the solvent control group.The NOEC and LOEC for hatching success were determined as 0.01 and 0.03 mg/L, respectively.
Larval survival until Day 30 post-hatch in the control and solvent control group was 95.0 and 93.75% thereby exceeding and satisfying the validity criteria for post-hatch survival (>75%).whereas in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas85.0, 72.50, 66.25, 53.75 and 25.0%,respectively. Larval survival reduction in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas 9.33, 22.67, 29.33, 42.67 and 73.33%,respectively in comparison with the solvent control group.In terms of concentrations, the NOEC and LOEC for post-hatch larval survival untilDay 30 were both considered to be equal to or less than 0.01 mg/L.
For fish total lengths, the NOEC and LOEC determined on Day 30 post-hatch were 0.01 and 0.03mg/L, respectively. For fish dry weights, the NOEC and LOEC determined on Day 30 post-hatch were 0.01 and 0.03 mg/L, respectively.
The concentrations of test chemical was determined using a validated liquid chromatographic method. Samples of each fresh test substance concentration, control and solvent control collected at exposure initiation and at renewals, and samples of each spent test substance concentration, control and solvent control collected at each renewal and at exposure termination were chemically determined.
In fresh samples of the test substance concentrations, the determined concentrations of test chemical was between 99.3 and 101.7%, concentrations, respectively. The results confirm that the test substance concentrations were within the acceptability range. In spent samples of the test substance concentrations, the determined concentrations of test chemical was between 96.1 and 96.9% of the concentrations, respectively.
All validity criteria were satisfied during the test, therefore the test was considered to be valid.
Results:The effects of test chemical on mortality, hatching success and fish larvae survivalDanio rerioin the definitive test are summarized in Table below.
LC50mortality |
0.064±0.01 |
EC50fertility |
0.34±0.047 |
EC50surviving |
0.31±0.06 |
||
NOECmortality |
<0.01 |
NOECfertility |
0.01 |
NOECsurviving |
<0.01 |
||
LOECmortality |
<0.01 |
LOECfertility |
0.03 |
LOECsurviving |
<0.01 |
||
Thus based on the results obtained chemical can be categorised as into chronic category 1 as per CLP classification criteria.
Key value for chemical safety assessment
Fresh water fish
Fresh water fish
- Effect concentration:
- 0.01 mg/L
Additional information
The aim of the study was todetermine the EC50, LOEC and NOECof test chemical on the growth and development of embryos and larvae of the freshwater fish species zebrafish (Danio rerio) in a Fish Early-Life Stage (ELS) Toxicity Test. The study was conducted according to the OECD guideline No. 210, Adopted 26thJuly, 2013.
The test was conducted with a semi-static design with renewal once 48 h.
Concentrated stock solutions and test media were analysed during the ELS test. Based on the range finding result the definitive test was conducted with five test concentrations of 0.01, 0.03, 0.10, 0.30 and 1.00 mg/L in terms of test chemical with a conversion factor of 3.2. Both control and solvent control were included in the test design. Each concentration was divided into four replicate with 20 eggs each and including the controls.
The cumulative mortality of the control and solvent control group at 30 days post-hatch was 11.25 and 11.25 %, whereas corrected cumulative mortality of embryos, larvae and juvenile fish after at the end of exposure to test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg /Lwas 12.68, 28.45, 49.30, 67.61and 92.96%,respectively.
Hatching success in the control and solvent control group was 93.42 and 94.67% respectively, satisfying the validation criterion for hatching success (70%),whereas in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas 91.18, 87.93, 67.92, 53.49and 25.00%,respectively. Fertility reduction in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas3.69, 7.12, 28.26, 43.50 and 73.59%,respectively in comparison with the solvent control group.The NOEC and LOEC for hatching success were determined as 0.01 and 0.03 mg/L, respectively.
Larval survival until Day 30 post-hatch in the control and solvent control group was 95.0 and 93.75% thereby exceeding and satisfying the validity criteria for post-hatch survival (>75%).whereas in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas85.0, 72.50, 66.25, 53.75 and 25.0%,respectively. Larval survival reduction in the group treated with test chemical at concentrations of 0.01, 0.03, 0.10, 0.30and 1.00 mg/Lwas 9.33, 22.67, 29.33, 42.67 and 73.33%,respectively in comparison with the solvent control group.In terms of concentrations, the NOEC and LOEC for post-hatch larval survival untilDay 30 were both considered to be equal to or less than 0.01 mg/L.
For fish total lengths, the NOEC and LOEC determined on Day 30 post-hatch were 0.01 and 0.03mg/L, respectively. For fish dry weights, the NOEC and LOEC determined on Day 30 post-hatch were 0.01 and 0.03 mg/L, respectively.
The concentrations of test chemical was determined using a validated liquid chromatographic method. Samples of each fresh test substance concentration, control and solvent control collected at exposure initiation and at renewals, and samples of each spent test substance concentration, control and solvent control collected at each renewal and at exposure termination were chemically determined.
In fresh samples of the test substance concentrations, the determined concentrations of test chemical was between 99.3 and 101.7%, concentrations, respectively. The results confirm that the test substance concentrations were within the acceptability range. In spent samples of the test substance concentrations, the determined concentrations of test chemical was between 96.1 and 96.9% of the concentrations, respectively.
All validity criteria were satisfied during the test, therefore the test was considered to be valid.
Results:The effects of test chemical on mortality, hatching success and fish larvae survivalDanio rerioin the definitive test are summarized in Table below.
LC50mortality |
0.064±0.01 |
EC50fertility |
0.34±0.047 |
EC50surviving |
0.31±0.06 |
||
NOECmortality |
<0.01 |
NOECfertility |
0.01 |
NOECsurviving |
<0.01 |
||
LOECmortality |
<0.01 |
LOECfertility |
0.03 |
LOECsurviving |
<0.01 |
||
Thus based on the results obtained chemical can be categorised as into chronic category 1 as per CLP classification criteria.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
