Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The reliability is rated Klimish 1

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
not specified
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Cement, alumina, chemicals
EC Number:
266-045-5
EC Name:
Cement, alumina, chemicals
IUPAC Name:
266-045-5
Details on test material:
- Name of test material (as cited in study report): Ciment Fondu®
- Substance type: mineral
- Physical state: dark grey fine powder
- Analytical purity : 100 %
- Purity test date: 2010-04-20
- Composition of test material, percentage of components: Ciment Fondu® is a mixture of more than 99.5% milled clinker of Ciment Fondu® which is the same substance as the registered substance here, and a grinding agent
- Lot/batch No.: 91478
- Storage condition of test material: at ambient temperature

Sampling and analysis

Analytical monitoring:
no
Details on sampling:
- Concentrations: 100 mg/l
- Sampling method: Dilution

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The 100 mg/l test solution was prepared by the addition of a nominal 2 g of test substance to 20 litres of dilution water. The test
solutions were then stirred overnight and then filtered using a pre-weighed 0.45 µm membrane filter. Post filtration, the membrane filters of the 100 mg/l test solution were oven dried and re-weighed in order to calculate the amount of compound removed during filtration. 1.39g of compound was removed during filtration, suggesting that 0.61g of Ciment Fondu® was dissolved in the media. The filtered solutions were clear and colourless. The 100 mg/l test solution required neutralisation using 1M HCL before the addition of the fish. The final 100 mg/l test solution was slightly cloudy

- Controls: The control solution was dilution water with no addition of test substance. The final dilution water control was clear and colourless

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Strain: Oncorhynchus mykiss
- Source: Houghton Springs Fish Farm, Winterbourne Houghton, Blandford Forum, Dorset DT11 0PD, UK
- Age at study initiation (mean and range, SD):
- Length: At the end of the exposure period, the fish from the dilution water control were measured. The range in length was 43 - 52 mm with a
mean of 48 mm. The loading of fish in the dilution water control was approximately 0.83 g/l
- Weight : At the end of the exposure period, the fish from the dilution water control were weighed. The range in weight was 1.01 - 2.15 g with a mean of 1.66 g. The loading of fish in the dilution water control was approximately 0.83 g l-1
- Method of breeding: the fish were held in 340 l fibreglass tanks under artificial lighting and were contained in water as described for the test
dilution water, except that the water was only filtered through a 10 µm filter before use. They were held at 15 +/- 1°C
Following their arrival on site the fish were held in 20% seawater as a prophylactic treatment, and additionally treated with twice daily prophylactic treatments of Formalin (25 ppm) and Malachite green (1 ml 100 l-1) for 9 days inclusive. The treatment was completed on 26 March 2010 after which
the fish were held in freshwater.
- Feeding during test : The fish were not fed during the course of the test


ACCLIMATION
- Acclimation period: for a minimum period of 7 days prior to the test
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: Fish were fed using a proprietary brand of high protein pelleted fish food
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): There were no mortalities recorded within the 48 hours prior to the start of the study

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
Hardness total as CaCO3 : 50.0 mg/l
Test temperature:
15 +/- 1 °C (Ranged from 14.4 to 15.2 °C) (Temperature values were determined daily using a thermometer calibrated to 0.1C and conforming to BS593)
T (°C) (Dilution water control) : 15.2 (0h) - 14.5 (24h) - 14.5 (48h) - 14.6 (72h) - 14.4 (96h)
T (°C) (100 mg/l Ciment fondu®) : 15.0 (0h) - 14.5 (24h) - 14.5 (48h) - 14.5 (72h) - 14.4 (96h)
pH:
pH values ranged from 7.49 to 8.04
Nota : pH of test solution was 9.21 prior to neutralisation with 1M HCl
pH (Dilution water control) : 7.96 (0h) - 7.61 (24h) - 7.49 (48h) - 7.60 (72h) - 7.70 (96h)
pH (100 mg/l Ciment fondu®) : 8.04 (0h) - 7.66 (24h) - 7.52 (48h) - 7.55 (72h) - 7.65 (96h)
Dissolved oxygen:
The dissolved oxygen concentrations (DO) in the test vessels ranged from 9.19 to 10.17 mg l-1 (91 to 101% of the air-saturation value)
DO (mg/l) (Dilution water control) (%) : 9.98 (99%) (0h) - 9.86 (98%) (24h) - 9.65 (96%) (48h) - 9.72 (97%) (72h) - 9.73 (97%) (96h)
DO (mg/l) (100 mg/l Ciment fondu®) (%) : 10.17 (101%) (0h) - 9.85 (98%) (24h) - 9.19 (91%) (48h) - 9.1 (90%) (72h) - 9.23 (92%) (96h)
Nominal and measured concentrations:
100 mg/l
Details on test conditions:
TEST SYSTEM


- Aeration: the test solutions were gently aerated
- Type of flow-through (e.g. peristaltic or proportional diluter): static
- No. of organisms per vessel: 10 fish were impartially selected and allocated to the test exposure concentration (100 mg/l) and the dilution water control


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water was tap water which had been coarsely filtered to remove particulate material, passed through activated carbon and dechlorinated using a sodium thiosulphate solution. Salts were added, as required, to maintain minimum hardness levels, and the treated water passed through an ultraviolet steriliser and filtered to 10 µm. The supply was then delivered to a temperature controlled header tank adjacent to the test laboratory which was set to the nominal test temperature of 15 ± 1°C, before distribution to the test rigs
- Particulate matter: < 3.00 mg/l solids suspended at 105 °C
- Pesticides: Ammoniacal nitrogen as N : < 2.00 µg/l - Organo-chlorine pesticides : < 0.05 µg/l - Organo-phosphorus pesticides : < 0.01 µg/l - PCB's : < 0.005 µg/l
- Chlorine (as Cl2) : Free residual : < 2 µg/l - Combined residual : < 2 µg/l
- Alkalinity (as CaCO3) : 37.6 mg/l
- Conductivity: 212 µS/cm (25 °C)


OTHER TEST CONDITIONS
- Adjustment of pH: The 100 mg/l test solution required neutralisation using 1M HCL before the addition of the fish
- Photoperiod: 16 hours fluorescent light and 8 hours dark with 20 minute dawn and dusk transition periods
- Light intensity: fluorescent light


TEST CONCENTRATIONS
- Spacing factor for test concentrations: no spacing factor, but one single concentration (100 mg/l)
- Justification for using less concentrations than requested by guideline:
- Test concentrations: 100 mg/l
- Results used to determine the conditions for the definitive study:
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
LC0
Effect conc.:
> 100 mg/L
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC0
Effect conc.:
> 100 mg/L
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LC0
Effect conc.:
> 100 mg/L
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
> 100 mg/L
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
> 100 mg/L
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: none
- Observations on body length and weight: none
- Other biological observations: none
- Mortality of control: none
- Other adverse effects control: darl discolouration, seen at 72h only

Any other information on results incl. tables

Sublethal observations / clinical signs:

Percentage Mortality :

 

Cumulative percentage mortality observed

   3 hours  24 hours 48 hours  72 hours  96 hours 
 Dilution water control  0
 

100 mg/l Nominal loading concentration

of Ciment Fondu®
Symptoms of toxicity observed :
            Symptoms observed   
   3 hours  24 hours 48 hours   72 hours 96 hours
 Dilution water control  A  A  A A  A
 100 mg/l Nominal loading concentration of Ciment Fondu®  A  A  A  C*  A
           

A

=

10% or less of test population dead or exhibiting symptoms of toxicity (considered not to be biologically significant).

 

B

=

Between 11% and 30% of test population dead or exhibiting symptoms of toxicity.

 

C

=

More than 30% of test population dead or exhibiting symptoms of toxicity.

 

*

=

Dark discoloured pigmentation

 

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Rainbow trout were exposed under static conditions for 96h to a single concentration of 100 mg/l of Ciment Fondu®. The 96h LC50 value is > 100 mg/l.
Executive summary:

In order to test acute toxicity of Ciment Fondu® to fish, according to OECD guideline 203, a group of ten Oncorrhynchus mykiss (Rainbow trout) was exposed under static conditions to the test solution of 100 mg/l nominal concentration of test substance and blank control solution (without test substance or any other additive) for a period of 96 hours.

The 100 mg/l test solution was prepared by the addition of a nominal 2 g of test substance to 20 litres of dilution water. The test

solutions were then stirred overnight and then filtered using a pre-weighed 0.45 µm membrane filter. The filtered solutions were clear and colourless. The 100 mg/l test solution required neutralisation using 1M HCL before the addition of the fish.

Mortalities and viable abnormalities were recorded at 3, 24, 48, 72 and 96 hours.

No mortality was recorded at any time.

The only general symptom of toxicity noted in this study was dark discolouration, which was seen at 72 hours only.

Conclusion : The 96h LC50 is > 100 mg/l.