Supporting information
Table SI. Rearing conditions and water chemistry for all tanks during the experimental period (34 days), showing the mean ± S.E. temperature
(Temp), salinity, isotope ratios and elemental ratios (element:43Ca)
Treatment
Rearing conditions
88
138
Temp Salinity Isotope Tank
Temp (°C)
Salinity
Sr:86Sr
Ba:137Ba
Sr:Ca (mmol mol-1)
Ba:Ca (µmol mol-1)
26°C
F
C
A
25.7 ± 0.2
0.0 ± 0.0
8.39 ±
2684 ± 953
386.90 ± 83.76
6.40 ± 0.26
0.17
C
B
25.7 ± 0.3
0.0 ± 0.0 8.42 ± 0.4 6.08 ± 0.47
3103 ± 1097
440.94 ± 181.43
W
A
27.1 ± 0.4
0.0 ± 0.0
1.59 ±
2460 ± 920
428.15 ± 126.30
1.12 ± 0.02
0.11
W
B
25.3 ± 0.3
0.0 ± 0.0
1.59 ±
2847 ± 1445
567.76 ± 350.62
1.14 ± 0.03
0.18
E
C
A
25.3 ± 0.3
19.0 ± 0.3
8.35 ±
10570 ± 1553
86.10 ± 5.00
6.47 ± 0.25
0.09
C
B
25.4 ± 0.2
19.0 ± 0.3 8.34 ± 0.1 6.42 ± 0.20
10002 ± 1004
53.07 ± 1.46
W
A
26.3 ± 0.2
20.0 ± 0.6
6.94 ±
14328 ± 1648
44.99 ± 7.22
0.84 ± 0.15
0.23
W
B
25.4 ± 0.2
20.0 ± 0.3
7.21 ±
9975 ± 695
44.33 ± 3.31
1.01 ± 0.04
0.07
M
C
A
26.7 ± 0.2
37.0 ± 0.8 8.3 ± 0.07 6.30 ± 0.21
12500 ± 2262
21.76 ± 5.38
C
B
26.8 ± 0.2
38.0 ± 0.6
8.33 ±
11752 ± 2501
20.41 ± 6.84
6.29 ± 0.28
0.05
W
A
26.7 ± 0.2
37.0 ± 0.9
7.64 ±
10990 ± 2141
25.30 ± 6.18
0.81 ± 0.06
0.06
W
B
26.9 ± 0.2
37.0 ± 0.8
7.69 ±
10579 ± 843
25.47 ± 5.16
0.81 ± 0.04
0.03
30°C
F
C
A
30.4 ± 0.1
0.0 ± 0.0
8.51 ±
1434 ± 894
246.47 ± 146.34
6.37 ± 0.39
0.53
C
B
30.8 ± 0.2
0.0 ± 0.0
8.74 ±
3057 ± 2385
364.00 ± 304.74
6.42 ± 0.34
0.24
W
A
30.2 ± 0.1
0.0 ± 0.0
3.47 ± 1.33 ± 0.05
4711 ± 1478
386.54 ± 198.09
1.49
W
B
31.1 ± 0.2
0.0 ± 0.0
1.71 ±
2303 ± 1067
424.88 ± 236.52
1.21 ± 0.03
0.16
E
C
A
30.6 ± 0.1
19.0 ± 0.3
8.28 ±
10336 ± 810
43.30 ± 2.75
6.39 ± 0.22
0.11
C
B
30.7 ± 0.2
20.0 ± 0.4
8.34 ±
11345 ± 2645
46.09 ± 14.40
6.54 ± 0.17
0.03
W
A
30.6 ± 0.1
20.0 ± 0.2
7.34 ±
8121 ± 929
44.73 ± 10.18
1.04 ± 0.07
0.12
W
B
30.7 ± 0.2
19.0 ± 0.2
7.21 ±
9074 ± 2366
46.22 ± 14.05
0.95 ± 0.02
0.01
M
C
A
30.4 ± 0.2
40.0 ± 0.5
8.26 ±
10385 ± 1387
18.01 ± 3.77
6.51 ± 0.12
0.11
C
B
30.6 ± 0.1
38.0 ± 0.8
8.35 ±
9882 ± 885
18.14 ± 4.92
6.39 ± 0.24
0.03
W
A
30.2 ± 0.2
40.0 ± 0.6
7.74 ±
11357 ± 4085
26.32 ± 10.93
0.89 ± 0.03
0.07
W
B
30.4 ± 0.1
38 .0± 0.9
7.71 ±
8667 ± 1816
21.77 ± 7.01
0.86 ± 0.05
0.03
F, fresh water (salinity 0); E, estuarine (salinity 20); M, marine (salinity 40); C, control treatment; W, water enriched with 137Ba and 86Sr.
Table SII. Analysis of variance of rearing conditions (recorded temperature and salinity)
among treatment tanks.
Temperature (T)
Salinity (S)
TS
Tank (T  S)
Residual
d.f.
1
2
2
6
448
Temperature
MS
F
2266.60 481.09
8.01
1.70
21.45
4.55
4.71
5.88
0.80
P
≤ 0.001
> 0.05
> 0.05
≤ 0.001
d.f.
1
2
2
6
352
Salinity
MS
F
25.77
2.25
44411.00 3873.00
12.65
1.10
11.47
3.20
3.59
P
> 0.05
≤ 0.001
> 0.05
< 0.05
DIET ISOTOPE ANALYSES
Experimental fish were fed on a marine-based 1.5 mm floating pellet diet (Lucky
Star) throughout the pre- and experimental periods. Triplicate diet samples were oven dried,
ground to a powder, dissolved in ultrapure HNO3 , diluted 50:1, and passed through a 0.45
µm filter (Woodcock et al., 2012). Samples were then analysed for Sr, Ba and Ca isotopes
following the same protocol as the water samples (refer to Materials and Methods).
The mean (± standard errors) element ratio were 88Sr:43Ca = 19.69 ± 0.73 mmol mol-1
and 138Ba:43Ca = 0.11 ± 0.001 mmol mol-1. Dietary isotope values were 88Sr/86Sr = 9.05 ±
0.06 and 138Ba/137Ba = 6.37 ± 0.01; however, these values were not used in the final
calculations of percent contribution values, as the isotopic values measured in the diet were
considerably higher than natural ratios (particularly for strontium).
Table SIII. Summary of literature assessing the per cent contribution of water to otolith Sr and Ba chemistry based on isotopic variations.
Adapted from Doubleday et al. (2013)
Species
Family
Environmental
Salinity
niche
Bidyanus bidyanus
Terapontidae
Obligate freshwater
0
Lates calcarifer
Latridae
Catadromous
0
Maccullochella peelii
Percichthyidae Obligate freshwater
0
Macquaria ambigua
Percichthyidae Obligate freshwater
0
Oreochromis niloticus
Cichlidae
Obligate freshwater
0
Oncorhynchus mykiss
Salmonidae
Anadromous
0
Salmo salar
Salmonidae
Anadromous
0
Acanthopagrus butcheri Sparidae
Obligate estuarine
10
Acanthopagrus butcheri Sparidae
Obligate estuarine
20
Lates calcarifer
Latridae
Catadromous
20
Acanthopagrus butcheri Sparidae
Obligate estuarine
30
Fundulus heteroclitus
Fundulidae
Opportunistic estuarine
30
Lates calcarifer
Latridae
Catadromous
40
a
ranges associated with different temperatures and within salinity treatments.
Isotope
type
enriched
enriched
enriched
enriched
radionatural
natural
enriched
enriched
enriched
enriched
enriched
enriched
%Sr
%Ba
71
77-93
64
64
88
66
30
67-78
76-81
65-99
62-83
83
82-84
92
82-89
89
87
73-82
69-76
64-75
70-78
98
64-76
Source
Doubleday et al. (2013)
this study a
Doubleday et al. (2013)
Doubleday et al. (2013)
Farrell and Campana (1996)
Gibson-Reinemer et al. (2009)
Kennedy et al. (2000)
Webb et al. (2012) a
Webb et al. (2012) a
this study a
Webb et al. (2012) a
Walther and Thorrold (2006)
this study a
Supporting Information References
Farrell, J.& Campana, S.E. (1996). Regulation of calcium and strontium deposition on the
otoliths of juvenile tilapia, Oreochromis niloticus. Comparative Biochemistry and
Physiology A 115, 103-109.
Gibson-Reinemer, D.K., Johnson, B.M., Martinez, P.J., Winkelman, D.L., Koenig, A.E.&
Woodhead, J.D. (2009). Elemental signatures in otoliths of hatchery rainbow trout
(Oncorhynchus mykiss): distinctiveness and utility for detecting origins and movement.
Canadian Journal of Fisheries and Aquatic Sciences 66, 513-524.
Walther, B.D. & Thorrold, S.R.( 2006). Water, not food, contributes the majority of strontium
and barium deposited in the otoliths of a marine fish. Marine Ecology Progress Series
311, 125-130.