Benzamide, N-[[4-[(cyclopropylamino)carbonyl]phenyl]sulfonyl]-2-methoxy-

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 Feb - 09 Mar 2005

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)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1400 (Fish Early-life Stage Toxicity Test)

Version / remarks:

(draft)

Qualifier:

according to guideline

Guideline:

other: FIFRA72-4

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Interval of analytical verification of test material: At experimental start, weekly (± 2 days) thereafter including experimental finish.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution was prepared with 208 mg a.i./L in 120 L stainless steel drums. The stock solution was mixed for 30 - 93 hours. The stock solution was used to dose the test aquaria via a modified proportional diluter system (Mount and Brungs, 1967). The temperature was approximately 20°C (room temperature) during mixing and use.
- Controls: Yes

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead Minnow
- Source: In-house fathead minnow breeding facility (Parental Lot Number: EG081104)

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish: 2 males and 5 females per aquaria
- Number of Spawning Substrates per Aquaria: 3
- Method of collection of fertilised eggs: Eggs were removed fromsubstrates contained in breeding tanks on the morning of the experimental start with nonviable eggs being discarded.

POST-HATCH FEEDING
- Start date: Day 6
- Type/source of feed: Brine shrimp (Artemia salind) starting on Day 4
- Amount given: Equal distribution at 0.5 - 3.0 mL per feeding
- Frequency of feeding: At least once daily on weekends and two to three times daily on weekdays until approximately 24 hours prior to study termination.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

35 d

Hardness:

soft water: 40 to 60 mg/L as CaCO3

Test temperature:

23.9 to 25.8 °C

pH:

7.6 to 8.3

Dissolved oxygen:

6.6 to 7.6 mg/L (80 to 92% saturation)

Nominal and measured concentrations:

Nominal Test Concentrations: Control, 0.63, 1.25, 2.50, 5.00 and 10.0 mg a.i./L
Measured Test Concentrations: Control (< 0.10), 0.70, 1.07, 2.58, 4.67 and 10.5 mg a.i./L

Details on test conditions:

TEST SYSTEM
- Emybro cups: One oscillating egg cup per replicate placed in each replicate through the egg hatching phase
- Test vessel:
- Material, size, fill volume: All Glass, 8.4-L, 7-L (21.6x12.7x25.5 cm) fill volume
- Type of flow-through: Modified Mount-Brungs proportional diluter system
- Renewal rate of test solution: Volume turnover per vessel per 24 hours aprox. 5 to 10
- No. of fertilized eggs/embryos per vessel: 35 eggs at initiation (Thinned to 20 alevin after hatching phase)
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 0.22 g /L static (mean wet weight based on controls), 0.09 g/L/day (mean biomass based on controls)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Process water (blended spring and reverse osmosis waters); according to guideline
- Culture medium different from test medium: No
- Intervals of water quality measurement: At experimental start, weekly (± 2 days) thereafter including experimental finish.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light, 8 hours dark (30 minutes light/dark transition period)
- Light intensity: 59.2 to 92.5 footcandles (637 to 996 lux)

EFFECT PARAMETERS MEASURED: Sublethal effects, fish hatchability, survival, and growth (length and dry weight- all surviving fish on Day 35)

POST-HATCH DETAILS
- Begin of post-hatch period: Day 6
- No. of hatched eggs (alevins)/treatment released to the test chamber: 20
- Release of alevins from incubation cups to test chamber on day no.: 5

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

10.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

4.67 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

length

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

4.67 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

weight

Table 1: Fish early life stage toxicity test

Test Object	Fathead Minnow
Exposure	35 Day, flow- through (ELS)
	NOEC	LOEC
Alevin Survival (Day 5):	10.5mga.i./L	>10.5mga.i./L
Fry Survival (Day 35):	10.5mga.i./L	>10.5mga.i./L
Percent Hatch:	10.5mga.i./L	>10.5mga.i./L
Time to Hatch:	10.5mga.i./L	>10.5mga.i./L
Growth (Length & Weight):	4.67 mga.i./L	10.5 mga.i./L
Morphological & Behavioral	10.5mga.i./L	>10.5mga.i./L
Observations:	No morphological or behavioral dose effects were apparent at any treatment group relative to the control group.
Conclusion:	The 35-day exposure to AE 0001789 technical resulted in a NOEC of 4.67 mg a.i./L and a LOEC of 10.5 mg a.i./L based on growth (length and weight).

 

 

 

Table 2: Measured Concentrations of AE 0001789 Technical During the Fathead Minnow Early Life Stage Toxicity Test

		Measured Concentration (mg a.i./L)
Nominal Conc. (mg a.i./L)	Replicate	Day 0	Day 7	Day 15	Day 21	Day 28	Day 35	Mean	Standard Deviation	Percent of Nominal
Control	A	<0.10	-	<0.10	-	<0.10	-	NA	NA	NA
	B	<0.10	-	<0.10	-	<0.10	-
	C	-	<0.10	-	<0.10	-	<0.10
	D	-	<0.10	-	<0.10	-	<0.10
0.63	A	0.72	-	0.73	-	0.70	-	0.70	0.03	110
	B	0.68	-	0.70	-	0.68	-
	C	-	0.73	-	0.72	-	0.69
	D	-	0.66	-	0.68	-	0.66
1.25	A	1.01	-	1.01	-	0.99	-	1.07	0.07	86
	B	1.03	-	1.08	-	1.06	-
	C	-	1.07	-	1.10	-	1.04
	D	-	1.16	-	1.20	-	1.14
2.50	A	2.57	-	2.59	-	2.52	-	2.58	0.03	103
	B	2.60	-	2.56	-	2.61	-
	C	-	2.57	-	2.61	-	2.56
	D	-	2.58	-	2.63	-	2.58
5.00	A	4.64	-	4.67	-	4.66	-	4.67	0.10	93
	B	4.59	-	4.62	-	4.68	-
	C	-	4.78	-	4.89	-	4.78
	D	-	4.60	-	4.60	-	4.58
10.0	A	10.4	-	10.4	-	10.2	-	10.5	0.16	105
	B	10.6	-	10.4	-	10.6	-
	C	-	10.6	-	10.7	-	10.4
	D	-	10.7	-	10.6	-	10.7

NA = Not applicable

LOQ = 0.10 mg/L

- Sample not collected

 

 

 

Table 3: Validity criteria for OECD 210

Criterion from the guideline	Outcome	Validity criterion fulfilled
The dissolved oxygen concentration should be >60% of the air saturation value throughout the test.	The dissolved oxygen concentration was with≥6.6 mg/L higher than 60% of the air saturation value throughout the test.	yes
The water temperature should not differ by more than ± 1.5 °C between test chambers or between successive days at any time during the test, and should be within the temperature ranges specified for the test species (Annex 2).	The water temperature did not differ by more than ± 1.5 °C between test chambers or between successive days at any time during the test.	yes
The analytical measure of the test concentrations is compulsory.	There is evidence that the concentration of the tested substance has been satisfactorily maintained, throughout the test (86 – 110% of nominal).	yes
Overall survival of fertilised eggs and post-hatch success in the controls and, where relevant, in the solvent controls should be greater than or equal to the limits defined in Annex 2.	Overall survival of fertilised eggs and post-hatch success in the controls was≥89%.	yes

 

Validity criteria fulfilled:

yes

Remarks:

See Table 3 in "Any other information on results incl. tables".

Description of key information

NOEC (35 d): 4.67 mg/L (arith. mean measured, length and weight, Pimephales promelas, freshwater, OECD 210)

 

NOEC (35 d): 10.5 mg/L (arith. mean measured, reproduction, Pimephales promelas, freshwater, OECD 210)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

NOEC

Effect concentration:

4.67 mg/L

Additional information

A 35-day early life stage test according to OECD 210 was conducted with the test substance under flow-through conditions on Fathead Minnow (Pimephales promelas, M-254985-01-1).

Fish were exposed to nominal test concentrations of 0.63, 1.25, 2.50, 5.00 and 10.0 mg a.i./L (measured: 0.70, 1.07, 2.58, 4.67 and 10.5 mg a.i./L), alongside a negative control.

The reported NOEC was 4.67 mg a.i./L for the parameter growth (length and weight) and the NOEC 10.5 mg a.i./L for reproduction (based on arith. mean measured concentrations). No morphological or behavioral effects for the test substance were apparent at any treatment group relative to the control group.