Exploration of Daphnia behavioural effects profiles induced by a broad
range of toxicants with different modes of action
Julie Chevalier, Elodie Harscoët, Merlin Keller, Pascal Pandard, Jérôme Cachot, Matthias
Grote
Supporting Information
a) Chemical tested
All chemicals products were purchased at VWR®, France. Esfenvalerate (CAS: 6623004-4), caffeine (CAS: 58-08-2), fipronil (CAS: 120068-37-3), cypermethrin (CAS: 52315-078), imidacloprid (CAS: 138261-41-3), sertraline (CAS: 79617-96-2) and copper sulphate
pentahydrate (CAS: 7758-99-8) were provided by as solid suspension of 250 g (purity 99%)
and of 25 mg for trichlorfon (CAS: 52-68-6) (purity 99%). Carbofuran (CAS: 1563-66-2) and
abamectin (CAS: 71751-41-2) were supplied as solution in pure methanol (1mL of 1g/L and
1mL of 100µg/mL respectively, purity 99%). Ethanol (CAS: 64-17-5), isopropanol (CAS: 6763-0) were supplied as pure solution (purity 99%). All tested solutions were prepared with
artificial reconstituted freshwater (6341 ISO, 2012). Since esfenvalerate, carbofuran and
abamectin have a low solubility in water, concentrated solution were prepared in methanol to
help solubilisation in water as it is recommended by OECD guidance document for difficult
substances (OECD, 2000) and was then diluted with ISO to obtain the percentage not
exceeding 0.01% in all treatments including control solvent.
b) Configuration of the test condition on the Bbe® Daphnia toximeter
For daphnids acclimation, an ISO water solution circulated with an optimal flowthrough of 33 mL/min at 20 ± 1 °C. After 2 hours, exposure was started by replacing the ISO
water with the test solution. This latter solution was placed in closed circuit after complete
ISO water evacuation (time previously estimated at 3 minutes by colorimetric measurement).
To be consistent with the immobilisation test, the experiments were conducted without food
supply in contrast to recommended test protocols for this system.
c) Average swimming speed ± standard error (per hour) of Daphnia magna
neonates exposed to several concentrations of (A) isopropanol (B) ethanol (C) copper
sulphate (D) trichlorfon (E) esfenvalerate (F) fipronil (G) sertraline (H) cypermethrin (I)
caffeine (J) abamectin (K) carbofuran and (L) imidacloprid for 48 hours in the “MultiDaphTrack” system.
7
(A)
6
5
4
3
2
1
0
0
10
20
30
40
50
7
(B)
average speed (mm/s)
6
5
4
3
2
1
0
0
10
20
30
40
50
7
(C)
6
5
4
3
2
1
0
0
10
20
30
Time (hours)
40
50
4
(D)
3
2
1
0
0
10
20
30
40
50
7
average speed (mm/s)
(E)
6
5
4
3
2
1
0
0
10
20
30
40
50
4
(F)
3
2
1
0
0
10
20
30
Time (hours)
40
50
5
(G)
4
3
2
1
0
0
10
20
30
40
50
7
(H)
average speed (mm/s)
6
5
4
3
2
1
0
0
10
20
30
40
50
5
(I)
4
3
2
1
0
0
10
20
30
Time (hours)
40
50
4
(J)
3
2
1
0
0
10
20
30
40
50
4
average speed (mm/s)
(K)
3
2
1
0
0
10
20
30
40
50
4
(L)
3
2
1
0
0
10
20
30
40
- – – – - –
50
Time (hours)
Controls
below EC5
near EC10
near EC20
EC50-EC70
>EC70
d) Daphnia magna chronic toxicity data from literature and on-line database
compared to the behavioral LOECs obtained from the “Multi-DaphTrack” system. (a:
(USEPA 2014), b: (Jemec et al. 2007), c: (Minagh et al. 2009), d: (Muyssen and Janssen
2007), e: (Firpo 2011), f: (Matsumoto 2009)).
Substance
Chronic test
Behavioral test
LOECs (mg/L)
NOECs (mg/L)
LOECs (mg/L)
Isopropanol
<1300
N.D.
N.D.
Ethanol
4800
16a
0.01a
Caffeine
25
N.D.
N.D.
Imidacloprid
85
1.5b
2.5b
Sertraline
0.03
0.032c
0.1c
Copper sulfate
<0.028
0.075d
0.09a
Fipronil
<0.001
0.010a
0.019a
Carbofuran
<0.011
0.01a
0.027a
Esfenvalerate
<0.0001
0.00011e
N.D.
Cypermethrin
0.00004
0.0000009a
0.000002a
Abamectin
<0.00004
0.00003a
0.00009a
Trichlorfon
0.00034
0.000006a
0.001f