Appendix S1.
Pelobates cultripes mainly breeds in large water bodies but also in small temporary ponds
(Diaz-Paniagua, 1990). The larvae of this species are much bigger than the ones from the other
species found in the Park. The larval period is long (Buchholz & Hayes, 2002; Gomez-Mestre &
Buchholz, 2006) so breeding takes place after the first rains in autumn and development may
extend for over six months (Diaz-Paniagua, Gomez-Rodriguez, Portheault et al., 2005). Triturus
pygmaeus is one of the most abundant species of Doñana. Their larval diet is largely comprised
by planktonic crustaceans (93%) and the larval period can last from February to the end of July
(Diaz-Paniagua et al. 2005). Discoglossus galganoi can breed several times in a year in shallow
puddles formed after heavy rains, or at the shallow edges of larger ponds. Larvae can be found
from December until May (Diaz-Paniagua et al. 2005). Bufo calamita lays large clutches in single
strings of eggs in a very short period following strong rains. Natterjacks prefer shallow pools
with scarce vegetation and sunny shores. Larvae are small and can be observed in Doñana ponds
from February. Hyla meridionalis lays eggs in small packages or individually and the larval
period takes approximately 2.5-3 months. Breeding often begins in January, and it goes on for
several months so larvae can be found in very different developmental stages in the temporary
ponds (Diaz-Paniagua et al. 2005). Pelophylax perezi starts breeding later than the other species,
generally from April. Larvae can be found mostly in the bottom feeding on detritus and in the
water column and they complete their development in the ponds during the summer or even
during the next year (Diaz-Paniagua et al., 2005).
References
Buchholz, D.R. & Hayes, T.B. (2002) Evolutionary patterns of diversity in spadefoot toad
metamorphosis (Anura : Pelobatidae). Copeia, 180-189.
Diaz-Paniagua, C. (1990) Temporary ponds as breeding sites of amphibians at a locality in
southwestern Spain. Herpetological Journal, 1, 447-453.
Diaz-Paniagua, C., Gomez-Rodriguez, C., Portheault, A. & De Vries, W. (2005) Los anfibios de
Doñana.
Gomez-Mestre, I. & Buchholz, D.R. (2006) Developmental plasticity mirrors differences among
taxa in spadefoot toads linking plasticity and diversity. Proceedings of the National
Academy of Sciences of the United States of America, 103, 19021-19026.
Table S1. Chemical parameters measured in tanks at the end of the experiment: Turbidity (NTU), Chlorophyll-a (μg L-1), Shannon Diversity Index
for the zooplankton (cladocerans, copepods, rotifers, and ostracods), total final plant biomass remaining in the tanks (g), total plant biomass after
re-flooding (g), dissolved oxygen in the water (mg L-1), conductivity (mS cm-1) and four dissolved nutrients (ammonium, phosphate, nitrate and
nitrite, ppb). Data represent mean ± se for every treatment (number of replicates per treatment, N=12, except for Low and NatC, N=11).
Treatment
No Amph
Low
High
No Pc
NatC
NatF
InvC
InvF
10.49 ± 4.16
38.43 ± 11.1
67.21 ± 18.87
4.85 ± 2.09
24.79 ± 5.32
27.79 ± 6.97
21.84 ± 8.42
116.17 ± 29.23
11.57 ± 3.23
11.30 ± 1.82
20.57 ± 4.33
16.55± 5.71
11.75 ± 1.40
13.64 ± 2.66
13.26 ± 2.79
20.22 ± 4.07
Zoo Shannon Div.
1.70 ± 0.1
1.51 ± 0.08
1.28 ± 0.06
1.56 ± 0.08
1.50 ± 0.15
1.47 ± 0.09
1.46 ± 0.12
1.57 ± 0.06
Total Plants (g)
Total Plants reflooding (g)
Diss. Oxygen (mg L-1)
404.34 ± 29.23
198.88 ± 37.06
58.69 ± 21.44
584.78 ± 75.51
262.03 ± 38.65
329.59 ± 48.95
261.78 ± 31.98
13.27 ± 3.91
80.10 ± 9.81
32.81 ± 7.46
19.71 ± 3.51
88.62 ± 7.87
24.56 ± 4.58
35.33 ± 9.16
30.93 ± 8.22
10.58 ± 2.21
8.57 ± 0.37
7.28 ± 0.37
6.47 ± 0.35
8.69 ± 0.35
7.67 ± 0.4
7.92 ± 0.39
7.53 ± 0.23
4.75 ± 0.24
Conductivity (mS cm-1)
0.25 ± 0.01
0.29 ± 0.01
0.33 ± 0.01
0.29 ± 0.02
0.30 ± 0.01
0.27 ± 0.01
0.29 ± 0.01
0.35 ± 0.02
Ammonium (ppb)
29.52 ± 4.76
71.32 ± 26.35
60.86 ± 14.41
26.98 ± 4.7
95.92 ± 51.08
47.86 ± 9.27
37.38 ± 6.86
240.4 ± 72.4
Phosphate (ppb)
20.95 ± 1.71
16.72 ± 2.1
20.78 ± 2.33
9.26 ± 0.94
22.68 ± 3.96
13.18 ± 2.09
15.14 ± 3.28
22.19 ± 3.08
Nitrate (ppb)
9.3 ± 2.81
10.96 ± 3.53
8.21 ± 1.53
11.1 ± 1.64
8.97 ± 1.26
6.87 ± 1.14
11.31 ± 2.45
9.61 ± 1.21
Nitrite (ppb)
4.13 ± 0.23
4.52 ± 0.66
7.64 ± 3.04
3.32 ± 0.17
10.9 ± 6.07
4.46 ± 1.21
3.83 ± 0.43
10.74 ± 3.08
Turbidity (NTU)
Chlorophyll-a (μg L )
-1
Table S2. Statistical results of the GLMM to test the effect of amphibians on different ecological
variables of temporary ponds. We used five different contrasts to test for such effects. Degrees of
freedom (df), F value (F) and p value (p) are shown for every contrast. Numerator df was equal to
1 in all statistical contrasts and therefore not shown in the table.
Effects of
amphibians on
temporary
ponds
Turbidity
Oxygen
Conductivity
Ammonium
Phosphate
Nitrite
Nitrate
Macrophytes
Macrophytes
after reflooding
Chlorophyll-a
Zooplankton
Shannon
Diversity
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
No Amph Low
No Amph -High
Low - High
No Amph No Pc
Low –
No Pc
86
10.85
0.0024
75
10.91
0.0015
86
5.95
0.031
78
2.89
0.0932
78
1.77
0.1871
78
0.02
0.889
78
2.26
0.137
86
14.95
0.00025
75
25.34
0.00021
78
0.39
0.5319
85
1.95
0.166
86
20.58
0.0002
75
27.38
0.0001
86
19.42
0.0005
78
1.59
0.2105
78
0.01
0.9075
78
0.39
0.5361
78
0.00
0.9997
86
64.84
0.00014
75
47.27
0.00021
78
4.84
0.0308
85
9.61
0.0026
86
1.31
0.2564
75
3.25
0.0755
86
3.5
0.0649
78
0.22
0.6415
78
1.48
0.2272
78
0.56
0.4576
78
2.26
0.1369
86
16.07
0.00014
75
2.81
0.0981
78
2.31
0.1329
85
2.67
0.1057
86
1.02
0.3157
75
0.1
0.7496
86
3.06
0.0838
78
0.67
0.416
78
8.07
0.017
78
2.22
0.1402
78
0.00
0.9997
86
4.42
0.0385
75
0.83
0.3644
78
0.39
0.5341
85
0.99
0.3223
86
18.33
0.0002
75
13.07
0.0006
86
0.53
0.4679
78
6.24
0.0029
78
8.07
0.017
78
1.73
0.1926
78
2.26
0.1369
86
35.08
0.00014
75
35.1
0.00021
78
0
0.9853
85
0.18
0.6731
Table S3. Statistical results of the GLMM to test the effect of native and invasive predators on
different ecological variables of temporary ponds. We used seven different contrasts to test for
such effects. Degrees of freedom (df), F value (F) and p value (p) are shown for every contrast.
Numerator df was equal to 1 in all statistical contrasts and therefore not shown in the table.
Effects of
predators
Turbidity
Oxygen
Conductivity
Ammonium
Phosphate
Nitrite
Nitrate
Macrophytes
Macrophytes
after
reflooding
Chlorophyll-a
Zooplankton
Shannon
Diversity
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
df
F
p
Low - NatC
Low - Nat F
Low - InvC
Low - InvF
86
0.57
0.4535
75
1
0.3198
86
0.1
0.7548
78
0.7
0.4047
78
1.19
0.2797
78
0.76
0.3852
78
2.16
0.1461
86
1.86
0.1763
75
1.12
0.2939
78
0.05
0.8307
85
0
0.9843
86
0.38
0.54
75
3.02
0.0866
86
1.07
0.3041
78
1.36
0.2464
78
1.8
0.1842
78
1.08
0.3012
78
0.02
0.8917
86
6.66
0.0128
75
0.01
0.928
78
0.17
0.6847
85
0.05
0.8287
86
3.24
0.0754
75
0.62
0.4338
86
0.03
0.8721
78
1.19
0.2781
78
0.88
0.3513
78
0.61
0.436
78
2.26
0.1369
86
2.36
0.1284
75
0.05
0.8325
78
0.05
0.8159
85
0.12
0.729
86
5.46
0.0218
75
35.89
0.0001
86
5.55
0.031
78
3.25
0.0752
78
1.68
0.1986
78
4.52
0.0366
78
2.26
0.1369
86
32.65
0.00014
75
11.5
0.00154
78
2.16
0.1461
85
0.25
0.6216
NatC NatF
86
0.02
0.8781
75
0.51
0.4794
86
1.83
0.1793
78
0.1
0.7571
78
6.02
0.0246
78
3.74
0.0567
78
1.87
0.1758
86
1.41
0.2384
75
1.38
0.2445
78
0.04
0.8513
85
0.04
0.8444
InvC InvF
86
17.89
0.0002
75
48.25
0.0001
86
5.04
0.0328
78
8.8
0.004
78
5.24
0.0297
78
8.89
0.0057
78
0.00
1.00
86
54.94
0.00014
75
10.61
0.002
78
1.6
0.2096
85
0.73
0.3957