Manganese sulphide

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-03-06 to 2010-04-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was conducted according to GLP and to current guidelines with no deficiencies.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Water samples were taken from the control and each replicate test vessel at 0 (fresh media), 24 and 96 hours (old media) for quantitative analysis.
Duplicate samples and samples at 24 (fresh media), 48 and 72 hours (fresh and old media) were taken and stored at approximately -20°C for further analysis if necessary.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: the test item was prepared as a saturated solution. An amount of test item (2100 mg) was added to 21 litres of dechlorinated tap water and stirred using a magnetic stirrer at approximately 100 rpm at approximately 14°C for 48 hours. This was prepared in triplicate. After stirring, any undissolved test item was removed by filtration through a 0.2 µm Sartorius Sartopore filter (first approximate 1 litre discarded to pre-condition the filter) and the filtrates pooled to give the 100% v/v saturated solution. Aliquots (2.0, 3.6, 6.4 and 11.2 litres) of the 100% v/v saturated solution were each separately diluted in a final volume of 20 litres of dechlorinated tap water, and stirred using a flat bladed mixer for approximately 1 minute, to give the remainder of the test concentrations of 10, 18, 32 and 56% v/v saturated solution respectively.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK
- Length at study initiation (length definition, mean, range and SD): 4.2 cm (sd = 0.1)
- Weight at study initiation (mean and range, SD): 0.95 g (sd = 0.07)
- Feeding during test: no

ACCLIMATION
- Acclimation period: 2010-03-17 to 2010-03-29
- Acclimation conditions: the same as test conditions
- Type and amount of food: commercial trout pellets
- Feeding frequency: daily
- Health during acclimation: no mortality observed during acclimation

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

no

Hardness:

140 mg/L as CaCO₃

Test temperature:

14°C

pH:

7.5 - 9.0

Dissolved oxygen:

8.7 - 12.1 mg O₂/L

Salinity:

not reported

Nominal and measured concentrations:

nominal concentrations: 10, 18, 32, 56 and 100% v/v saturated solution
measured concentrations: 1.3, 2.3, 4.0, 7.8 and 13 mg/L as test item

Details on test conditions:

TEST SYSTEM
- Test vessel: 20 litre glass exposure vessels
- Type: open, covered to reduce evaporation
- Aeration: yes, test vessels aerated via narrow bore glass tubes
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Biomass loading rate: 0.29 g/L

TEST MEDIUM / WATER PARAMETERS
- Source: ZDB13 ALVASTON
- Preparation of dilution water: ALVASTON tap water was dechlorinated by passage through an activated carbon filter (Purite Series 500) and partly softened (Elga Nimbus 1248D Duplex Water Softener).

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods

EFFECT PARAMETERS MEASURED: Mortalities and any sub-lethal effects were observed 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.7 - 1.8
- Range finding study: yes, two range finding tests were performed. In the initial range-finding test fish were exposed to a maximum measured test concentration of 12.2 mg/L as test item and in the second range-finding test fish were exposed to test concentrations of 1.0, 10 and 100% v/v saturated solution.
- Test concentrations: In the definitive test concentrations of 10, 18, 32, 56 and 100% v/v saturated solution were assigned.
- Results used to determine the conditions for the definitive study: The results of the second range finding test showed a single mortality at the test concentration of 100% v/v saturated solution. No mortalities were observed at the test concentrations of 1.0 and 10% v/v saturated solution.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

56 other: % v/v saturated solution

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 13 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

7.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The results showed 100% mortalities at the test concentration of 100% v/v saturated solution in the initial range-finding test (results are given in Table 1). There were no sub-lethal effects of exposure during the initial range-finding test.
Analysis of the 100% v/v saturated solution showed a measured concentration of 12.2 mg/L as test item was obtained.

A single mortality was observed in the second range-finding test at the test concentration of 100% v/v saturated solution. No mortalities were observed at the test concentrations of 1.0 and 10% v/v saturated solution. Cumulative mortality data from the exposure of rainbow trout to the test item during the second range-finding test are given in Table 2 and no sub-lethal effects of exposure are given in Table 3.

The results of the definitive test showed the highest mean measured test concentration resulting in 0% mortality to be 7.8 mg/L, the lowest mean measured test concentration resulting in 100% mortality to be greater than 13 mg/L and the No Observed Effect Concentration (NOEC) to be 7.8 mg/L.The No Observed Effect Concentration based on nominal test concentrations was 56% v/v saturated solution. The No Observed Effect Concentration is based upon zero mortalities and the absence of any sub-lethal effects of exposure at this concentration. Cumulative mortality data from the exposure of rainbow trout to the test item during the definitive test are given in Table 4.

A single fish in the control group was lost after 72 hours having jumped out of the tank despite the exposure vessel being covered. This was considered not to affect the outcome of the test as no adverse effects were observed in the control group.

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

The test preparations were observed to be clear, colourless solutions throughout the duration of the test.

Results with reference substance (positive control):

not reported

Reported statistics and error estimates:

not reported

Sublethal observations / clinical signs:

Table1            Cumulative Mortality Data in the Initial Range-finding Test

Nominal Concentration (% v/v Saturated Solution)	Cumulative Mortality (Initial Population = 3)
	3 Hours	6 Hours	24 Hours	48 Hours	72 Hours	96 Hours
Control	0	0	0	0	0	0
100	0	0	0	2	3	3

Table2            Cumulative Mortality Data in the Second Range-finding Test

Nominal Concentration (% v/v Saturated Solution)	Cumulative Mortality (Initial Population = 3)
	3 Hours	6 Hours	24 Hours	48 Hours	72 Hours	96 Hours
Control	0	0	0	0	0	0
1.0	0	0	0	0	0	0
10	0	0	0	0	0	0
100	0	0	0	1	1	1

Table3            Sub-lethal Effects of Exposure in the Second Range-finding Test

Nominal Concentration (% v/v Saturated Solution)	Sub-lethal Effects	Time (Hours)
		3	6	24	48	72	96
Control	No abnormalities detected
1.0	No abnormalities detected
10	No abnormalities detected
100	Increased pigmentation					2/2	2/2

Table4            Cumulative Mortality Data in the Definitive Test

Mean Measured Concentration (mg/L as Test Item)	Cumulative Mortality (Initial Population = 7)						% Mortality
	3 Hours	6 Hours	24 Hours	48 Hours	72 Hours	96 Hours	96 Hours
Control	0	0	0	0	1[1]	1	14
1.3	0	0	0	0	0	0	0
2.3	0	0	0	0	0	0	0
4.0	0	0	0	0	0	0	0
7.8	0	0	0	0	0	0	0
13	0	0	0	0	2	2	29

[1]A single fish in the control group was lost after 72 hours having jumped out of the tank despite the exposure vessel being covered. This was considered not to affect the outcome of the test as no adverse effects were observed in the control group.

Appendix4      Physico-Chemical Measurements

Mean Measured Concentration (mg/L as Test Item)	Time (Hours)
	0 Hours (Fresh Media)				24 Hours (Old Media)				24 Hours (Fresh Media)
	pH	mg O2/L	%ASV*	TºC	pH	mg O2/L	%ASV*	T°C	pH	mg O2/L	%ASV*	T°C
Control	7.8	9.9	96	14	8.1	9.5	92	14	7.7	11.4	111	14
1.3	7.5	9.9	96	14	8.1	9.4	91	14	7.6	11.1	108	14
2.3	7.5	10.0	97	14	8.2	9.8	95	14	7.5	11.2	109	14
4.0	7.5	9.6	93	14	8.0	9.0	87	14	7.6	11.1	108	14
7.8	8.2	10.0	99	15	8.2	9.3	90	14	7.9	11.0	109	15
13	8.9	9.2	91	15	8.3	8.9	86	14	8.6	10.5	104	15

 

Mean Measured Concentration (mg/L as Test Item)	Time (Hours)
	48 Hours (Old Media)				48 Hours (Fresh Media)				72 Hours (Old Media)
	pH	mg O2/L	%ASV*	TºC	pH	mg O2/L	%ASV*	T°C	pH	mg O2/L	%ASV*	T°C
Control	8.0	9.0	87	14	7.8	10.6	103	14	8.3	9.3	90	14
1.3	8.3	9.7	94	14	7.9	10.8	103	13	8.2	9.1	88	14
2.3	8.3	9.8	95	14	7.9	10.6	103	14	8.1	9.4	91	14
4.0	8.2	9.1	88	14	7.9	10.3	100	14	8.0	9.1	88	14
7.8	8.3	9.4	91	14	8.0	9.3	90	14	8.4	9.3	90	14
13	8.4	9.8	95	14	8.9	8.4	82	14	8.7	9.5	92	14

 

Mean Measured Concentration (mg/L as Test Item)	Time (Hours)
	72 Hours (Fresh Media)				96 Hours (Old Media)
	pH	mg O2/L	%ASV*	TºC	pH	mg O2/L	%ASV*	T°C
Control	8.2	10.3	100	14	8.1	11.6	113	14
1.3	8.0	10.9	106	14	8.2	11.9	116	14
2.3	8.0	10.7	104	14	8.2	11.6	113	14
4.0	8.1	10.7	104	14	8.3	12.1	117	14
7.8	8.4	9.4	91	14	8.2	12.1	117	14
13	9.0	8.7	86	15	8.2	11.9	116	14

 

*ASV= Dissolved oxygen concentration expressed as a percentage of Air Saturation Value

Validity criteria fulfilled:

yes

Conclusions:

96-Hour LC50 was greater than 13 mg/L.

Executive summary:

The short-term toxicity of the test material to fish was assessed according to OECD Test Guideline 203 and EU Method C.1 and in compliance with GLP.

The results showed 100 % mortalities at the test concentration of 100 % v/v saturated solution in the initial range-finding test. There were no sub-lethal effects of exposure during the initial range-finding test.

Analysis of the 100 % v/v saturated solution showed a measured concentration of 12.2 mg/L as test material was obtained.

A single mortality was observed in the second range-finding test at the test concentration of 100 % v/v saturated solution. No mortalities were observed at the test concentrations of 1.0 and 10 % v/v saturated solution.

The results of the definitive test showed the highest mean measured test concentration resulting in 0 % mortality to be 7.8 mg/L, the lowest mean measured test concentration resulting in 100 % mortality to be greater than 13 mg/L and the No Observed Effect Concentration (NOEC) to be 7.8 mg/L. The No Observed Effect Concentration based on nominal test concentrations was 56 % v/v saturated solution. The No Observed Effect Concentration is based upon zero mortalities and the absence of any sub-lethal effects of exposure at this concentration.

A single fish in the control group was lost after 72 hours having jumped out of the tank despite the exposure vessel being covered. This was considered not to affect the outcome of the test as no adverse effects were observed in the control group. There were no sub-lethal effects of exposure observed in the test.

Under the conditions of the study the 96-Hour LC50 was greater than 13 mg/L.

Description of key information

The 96-Hour LC50 was greater than 13 mg/L.

Additional information

The short-term toxicity of the test material to fish was assessed according to OECD Test Guideline 203 and EU Method C.1 and in compliance with GLP.

The results showed 100 % mortalities at the test concentration of 100 % v/v saturated solution in the initial range-finding test. There were no sub-lethal effects of exposure during the initial range-finding test.

Analysis of the 100 % v/v saturated solution showed a measured concentration of 12.2 mg/L as test material was obtained.

A single mortality was observed in the second range-finding test at the test concentration of 100 % v/v saturated solution. No mortalities were observed at the test concentrations of 1.0 and 10 % v/v saturated solution.

The results of the definitive test showed the highest mean measured test concentration resulting in 0 % mortality to be 7.8 mg/L, the lowest mean measured test concentration resulting in 100 % mortality to be greater than 13 mg/L and the No Observed Effect Concentration (NOEC) to be 7.8 mg/L. The No Observed Effect Concentration based on nominal test concentrations was 56 % v/v saturated solution. The No Observed Effect Concentration is based upon zero mortalities and the absence of any sub-lethal effects of exposure at this concentration.

A single fish in the control group was lost after 72 hours having jumped out of the tank despite the exposure vessel being covered. This was considered not to affect the outcome of the test as no adverse effects were observed in the control group. There were no sub-lethal effects of exposure observed in the test.

Under the conditions of the study the 96-Hour LC50 was greater than 13 mg/L.