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Diss Factsheets

Ecotoxicological information

Long-term toxicity to fish

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Administrative data

Endpoint:
fish early-life stage toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
ZO-1 / BON / BNE
IUPAC Name:
ZO-1 / BON / BNE
Test material form:
other: solution in dimethylformamide
Details on test material:
- Name of test material (as cited in study report): ZO-1
- Substance type: organic
- Physical state: powder
- Lot/batch No.: 0603
- Storage condition of test material: at room temperature in the dark

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations:
- Sampling method: Water samples were taken from the solvent control and all surviving test groups (replicates R1 and R2 pooled)
on days 0, 3, 6, 10, 13, 17, 20, 24, 27 and 31 (fresh media) and days 1, 4, 7, 11, 14, 18, 21, 25, 28 and 32 (old media) for quantitative analysis.
Samples were anylysed untreated and after centrifugation.
- Sample storage conditions before analysis: Duplicate samples were taken and stored frozen (approx. -20 °C) for further analysis if necessary.

Test solutions

Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: without dimethylformamide and with 100 µl/l dimethylformamide
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethylformamide
- Concentration of vehicle in test medium
stock solution: ~100% and
final test solutions including solvent control: 100 µl/l


Test organisms

Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: fathead minnow (Pimephales promelas)
- Source: Osage Catfisheries Inc., Osage beach, Missouri, USA

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish (i.e. of females used to provide required number of eggs):
- Method of collection of fertilised eggs: Each breeding tank was supplied with inverted plastic guttering for the fish to lay eggs on and be fertilised.
Fertilised eggs were collected from the breeding tanks. The eggs were less than 24 h oold on introduction into the test system.
- Subsequent handling of eggs:

POST-HATCH FEEDING
- Start date:
- Type/source of feed:
- Amount given:
- Frequency of feeding:

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
32 d
Post exposure observation period:
28 days post hatch

Test conditions

Hardness:
118 - 138 mg/l as CaCO3
Test temperature:
approx. 25 °C
Dissolved oxygen:
greater than or equal to 8.3 mg O2/l
Nominal and measured concentrations:
nominal concentrations: 0.0027, 0.0048, 0.0084, 0.015 and 0.027 mg/l
time weighted mean measured concentrations: 0.000959, 0.00157, 0.00287, 0.00512 and 0.0109 mg/l
percentage of nominal test concentration: 36, 33, 34, 34 and 41 %
Details on test conditions:
TEST SYSTEM
- Emybro cups (if used, type/material, size, fill volume):
- Test vessel:
- Type (delete if not applicable): open / closed
- size: 12 l,
fill volume: 10 l
- Aeration: via narrow bore glass tubes
- Renewal rate of test solution (frequency): daily
- No. of fertilized eggs per vessel: 30
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory tap water dechlorinated by passage through an activated carbon filter (Elga AC1)
and partly softened (Elga Nimbus 1248 Duplex Water Softener) giving water with a total hardness of approx. 100 mg/l as CaCO3
- Total organic carbon: 1.7 mg C/l (prior to dechlorination and softening)
- Intervals of water quality measurement: water temperature, pH, light intensity and dissolved oxygen concentrations were recorded daily throughout the test

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h darkness with 20 min dusk and dawn transition periods

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
- Test concentrations (nominal): 0.0027 and 0.027 mg/l (water solubility limit)
- Results used to determine the conditions for the definitive study: 93 % mortality at 0.027 mg/l

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):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
32 d
Dose descriptor:
LOEC
Effect conc.:
0.008 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
weight
Duration:
32 d
Dose descriptor:
LOEC
Effect conc.:
0.003 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
weight
Duration:
32 d
Dose descriptor:
NOEC
Effect conc.:
0.005 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
weight
Duration:
32 d
Dose descriptor:
NOEC
Effect conc.:
0.002 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
weight
Duration:
32 d
Dose descriptor:
NOEC
Effect conc.:
0.005 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
length
Duration:
32 d
Dose descriptor:
NOEC
Effect conc.:
0.002 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
length
Details on results:
- Mortality at egg, larval and adult stages: refer to Tables under Attached Backgroud Material
6 Number of Dead Eggs in the Definitive Test
7 Number of Dead Larvae in the Definitive Test
- Days to hatch or time to release of young: start of egg hatching on Day 3
completion of hatching on Day 4
- Numbers hatched: refer to Tables under Attached Backgroud Material
8 Cumulative Number of Hatched (Live) Larvae in the Definitive Test
9 Number of Hatched (Live) Larvae (Non-cumulative) in the Definitive Test
- Number of fish in swim-up stage at one or more time periods (e.g., day x1, x2):
- Observations on body length and weight of young and/or exposed parents at one or more time periods:


- mean hatching rate: 92 - 100 % for all concentrations
The test material did not affect the hatching success of the eggs.
- mean survival rate: 79 - 93 % for all concentrations except 0.027mg/l
14 % for concentration 0.027 mg/l: prolonged toxic effect on the survival of the larvae at 0.027 mg/l
For more details please refer to Table 10 under Attached Background Material
Reported statistics and error estimates:
For the estimation of the LOEC and the NOEC the length and dry weight data obtained on termination of the test for the solvent control and each test group were compared using
one way analysis of variance incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981) and
Dunnett's multiple comparison procedure for comparing several treatments with a control (Dunnett, 1955).
All statistical analyses were performed using the SAS computer software package (SAS, 1999 - 2001).

Any other information on results incl. tables

 

Nominal Test Concentration (mg/l)

 

 

 

Control

Solvent Control

0.0027

0.0048

0.0084

0.015

0.027

Body Length
(mean ± standard deviation, mm)

15.5±1.7

15.5±1.3

15.2±1.7

15.2±1.5

14.7±1.7

13.7±1.3

13.4±1.0

Body Weight
(mean ± standard deviation, mg)

14.9±5.1

13.2±4.4

13.2±4.6

12.2±4.0

11.0±4.0

8.8±3.0

7.8±1.8

Fish length and dry weight data at termination of the test

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Executive summary:

The Lowest Observed Effect Concentration was considered to be

0.0084 mg/l (0.0029 mg/l based on the time-weighted mean measured test concentrations of the centrifuged test media)

on the basis that at this test concentration significant differences (P<0.05) were observed between the solvent control and the 0.0084 mg/l test group in terms of fish weight.

The No Observed Effect Concentration was considered to be

0.0048 mg/l (0.0016 mg/l based on the time-weighted mean measured test concentrations of the centrifuged test media)

on the basis that at this test concentration there were no significant differences (P2:0.05) in terms of both fish length and weight when compared to the solvent control and that there was no effect on hatching and survival rates.

There were no sub-lethal effects observed in the test.