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Ecotoxicological information

Long-term toxicity to fish

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Reference
Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 215 (Fish, Juvenile Growth Test)
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Water samples were taken from the control and all surviving test groups on days 0, 4, 7, 11, 14, 18, 21 and 27 (fresh media) and on days 1, 5, 8, 12, 15, 19, 22 and 28 (old media) for quantitative analysis.
- Sample storage conditions before analysis: Duplicate samples were taken and stored at approximately - 20°C for further analysis if necessary.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION:
- Method: Amounts of test material (4.00, 7.20, 12.80, 22.40 and 40.00 g) were separately dispersed in 11 litres of dechlorinated tap water with the aid of a propeller stirring at approxiamtely 1500 rpm at 14°C for approximately 2 hours. After stirring each of the mixtures were then separately dispersed in a final volume of 30 litres of dechlorinated tap water to give the 100, 180, 320, 560 and 1000 mg/l test concentrations.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: Rainbow trout
- Source: Brow Well Fisheries Limited, Yorkshire, UK
- Weight at study initiation (mean, SD): 2.47 g (sd = 0.27 g)


FEEDING DURING TEST
- Food type: Commercial trout pellets
- Amount: 4% wet bodyweight total per day
- Frequency: Yes –fed daily in two equal portions per day.
Test type:
semi-static
Limit test:
no
Total exposure duration:
28 d
Hardness:
156 mg/ as CaCO3
Test temperature:
Refer to attached document for individual measurements
pH:
The pH of monosodium cyanurate monohydrate is 9.0-9.5 (1% slurry). Therefore the pH of the fresh media increased slightly at higher test concentrations. The initial pH ranged from7.5 to 8.6. THe pH range narrowed as the media aged, because the test media slowly equilibrated with atmospheric carbon dioxide due to aeration. By the time the media were replaced after approximately 24 hours, the pH range had narrowed to 7.3 to 8.3. (Refer to attached document for individual measurements)
Dissolved oxygen:
Refer to attached document for individual measurements
Nominal and measured concentrations:
100, 180, 320, 560 and 1000 mg/L (equivalent to 76, 136, 242, 423 and 756 mg cyanuric acid/L)
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type: open (test vessels covered to reduce evaporation)
- Material, size, headspace, fill volume: 40 L glass vessels
- Aeration: Test vessels aerated via narrow bore glass tubes
- Type of flow-through: Single pass renewal system
- Renewal rate of test solution: Daily renewal of test preparations
- No. of organisms per vessel: 16
- No. of vessels per concentration (replicates):1
- No. of vessels per control (replicates): 1
- Biomass loading rate:0.99 g bw/l


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated laboratory tap water
- Total organic carbon: Less than the limit of quantitation
- Particulate matter: 0.3 mg/l
- Culture medium different from test medium: No
- Intervals of water quality measurement: Water temperature and dissolved oxygen content recorded daily


OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h darkness with 20 minute dawn and dusk transition time


EFFECT PARAMETERS MEASURED: Mortalities and sub-lethal effects recorded daily. Fish weight measured on day 0 and 28


VEHICLE CONTROL PERFORMED: no


RANGE-FINDING STUDY
- Test concentrations: 10, 100 and 1000 mg/l
- Results used to determine the conditions for the definitive study: Based on the results the range finding tests the following test concentrations were assigned for the definitive test: 100, 180, 320, 560 and 1000 mg/l
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: Equivalent to 756 mg/L cyanuric acid
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: Equivalent to 756 mg/L cyanuric acid
Details on results:
- Fish weights (individual and mean values) on day 0 and 28: see Table 1 and 2
- Tank-average or pseudo-specific growth rates for periods 0-28: see Table 3

Verification of test concentrations:

Analysis of the unfiltered test preparations (both fresh and old media) throughout the duration of the study showed the measured test concentrations to range from 97% to 106% of nominal with the exception of the 560 and 1000 mg/l test concentrations on day 21 (fresh media) which showed measured test concentrations of less than LOQ and 137% of nominal respectively. These were considered to be erroneous therefore frozen duplicate samples were analyzed. This analysis showed measured test concentrations of 98% of nominal for both the 560 and 1000 mg/l test concentrations indicating the true dosed concentrations and confirming the original analysis to be erroneous. Analysis of the filtered test preparations ( both fresh and old media) throughout the duration of the test showed the measured test concentrations to range from 97% to 104% of nominal and so the results are based on nominal test concentrations only.

Table 1: Individual fish weight data (g) - day 0 values

Fish

Nominal concentration

(mg/l)

Control

100

180

320

560

1000

1

2.70

2.22

2.72

2.98

2.49

2.32

2

2.07

2.84

2.23

2.85

2.36

2.44

3

2.20

2.58

2.17

2.72

2.74

2.84

4

2.12

2.62

2.41

2.58

2.68

2.40

5

2.28

2.67

2.66

2.83

2.21

2.08

6

2.69

2.16

2.57

2.90

2.92

2.57

7

2.58

2.33

2.86

2.09

2.93

2.88

8

2.84

2.17

2.84

2.19

2.42

2.05

9

2.59

2.42

2.77

2.51

2.85

2.24

10

2.78

2.38

2.94

2.53

2.67

2.70

11

2.60

2.32

2.42

2.08

2.06

2.04

12

2.35

2.45

2.16

2.70

2.69

2.62

13

2.36

2.65

2.67

2.42

2.48

2.15

14

2.83

2.87

2.08

2.20

2.45

2.49

15

2.10

2.47

2.26

2.13

2.38

2.12

16

2.45

2.12

2.56

2.13

2.72

2.90

Mean

2.47

2.45

2.52

2.49

2.57

2.43

SD

0.27

0.23

0.28

0.32

0.25

0.30

Table 2: Individual fish weight data (g) - day 28 values

Fish

Nominal concentration

(mg/l)

Control

100

180

320

560

1000

1

4.03

5.49

6.51

6.13

6.02

4.71

2

4.54

6.84

6.30

6.65

5.90

5.45

3

5.70

4.88

6.72

4.94

5.76

5.49

4

5.08

6.60

4.82

6.38

4.34

4.90

5

4.75

6.03

5.63

6.27

6.19

4.39

6

4.55

6.46

6.16

4.59

5.59

6.87

7

4.44

5.33

8.00

6.52

5.03

4.99

8

4.19

6.42

5.25

5.26

6.62

5.15

9

5.13

6.18

4.97

4.31

6.25

6.21

10

4.35

5.54

5.36

7.03

6.85

7.05

11

5.34

5.45

6.79

5.82

6.90

5.70

12

3.86

5.89

6.57

6.04

6.25

5.76

13

5.61

6.23

5.80

4.45

5.71

6.97

14

4.04

5.70

5.32

6.75

5.25

5.35

15

5.56

4.86

5.61

5.01

6.04

5.04

16

-

5.59

5.01

7.84

5.80

6.47

Mean

4.74

5.84

5.93

5.87

5.91

5.66

SD

0.62

0.59

0.85

1.02

0.66

0.83

Table 3: Tank average specific growth rates

Nominal concentration (mg/l)

Tank-average specific growth rates

(0-28 days)

Control

2.321

100

3.096

180

3.042

320

3.041

560

2.972

1000

3.011

Validity criteria fulfilled:
yes
Conclusions:
Exposure of juvenile trout to the test material for a period of 28 days resulted in no significant inhibition of the tank-average specific growth rate at the nominal test concentrations of 100, 180, 320, 560 and 1000 mg/L respectively over the test period of 0 to 28 days when compared to the control group.
One fish was killed from the control group on day 18 as a precautionary measure. All other fish survived the 28 days test period.
There were no sublethal effects of exposure observed in the test. The NOEC was equivalent to 756 mg of CYA/L and the LOEC was equivalent to > 756 mg of CYA/L.

Description of key information

The chlorinated isocyanurates are unstable in the environment, because the free available chlorine is rapidly reduced.  CYA, or its salt, is the stable degradation product. Therefore, CYA, or its sodium salt, is the substance of interest for chronic ecotoxicity studies. A fish juvenile growth test with rainbow trout was performed with the monosodium salt of cyanuric acid (equivalent to 75.6% cyanuric acid) according to OECD 215.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
756 mg/L

Additional information

A fish juvenile growth test with rainbow trout was performed with the monosodium salt of cyanuric acid (equivalent to 75.6% cyanuric acid) according to OECD guideline 215. Fish were exposed for 21 days to nominal concentrations of 100, 180, 320, 560 and 1000 mg/L (equivalent to 76, 136, 242, 423 and 756 mg cyanuric acid/L).

Zero mortalities, no inhibition of tank average specific growth rate, no sublethal effects of exposure and no significant reduction in terms of the “pseudo” specific growth rate were observed when compared to the control group. The NOEC for cyanuric acid was determined to be 756 mg/L