Diversity 2016, 8, 9; doi:10.3390/d8020009
S1 of S7
Supplementary Materials: Detection of a
High-Density Brachiolaria-Stage Larval Population of
Crown-of-Thorns Sea Star (Acanthaster planci) in
Sekisei Lagoon (Okinawa, Japan)
Go Suzuki, Nina Yasuda, Kohta Ikehara, Kouki Fukuoka, Takahiko Kameda, Sayaka Kai,
Satoshi Nagai, Atushi Watanabe, Takashi Nakamura, Shunsuke Kitazawa,
Lawrence Patrick C. Bernardo, Tatsuya Natori, Momoka Kojima and Kazuo Nadaoka
Table S1. Number of larvae isolated under a microscope (n = 225) and larvae identified using
polymerase chain reaction (PCR) and sequencing analyses (n = 42 in total).
17-Jun
SS02
SS03
SS05
SS07
SS10
SS11
SS13
SS15
SS17
SS19
SS20
SS21
SS26
SS27
SS28
SS30
SS31
SS32
SS33
SS34
SS35
Number of Larvae
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
Isolated
Identified as A. planci
18-Jun
First screening Gastrula Bipinnaria Brachiolaria
First screening Gastrula Bipinnaria
21-Jun
Brachiolaria First screening Gastrula Bipinnaria Brachiolaria
6
0
9
0
2
0
5
1
○
1
0
○
○
3
3
3
1
1
0
2
2
1
0
5
1
68
16
2
0
1
1
2
0
○
3
2
1
1
3
0
3
0
2
0
30
8
3
0
17
0
2
0
3
1
7
0
13
2
5
0
5
0
○
○
○
1
0
1
1
2
0
1
0
1
1
1
1
○
○
○
1
0
3
0
5
0
1
0
Diversity 2016, 8, 9; doi:10.3390/d8020009
S2 of S7
Table S2. The most closely related species obtained from larval DNA other than Acanthaster planci
based on a BLAST search using the partial cytochrome oxidase subunit 1 (CO1) sequence. GS:
gastrula, BP: bipinnaria, BR, brachiolaria. These stages were identified using echinoderm universal
CO1 primers following the protocol of Arndt et al. (1996) [1].
Individual number Site
105 SS17
1 SS34
141 SS20
158 SS20
108 SS17
29 SS17
39 SS17
174 SS27
34 SS17
12 SS20
42 SS17
182 SS34
89 SS17
97 SS15
99 SS15
16 SS17
20 SS17
23 SS17
30 SS17
40 SS17
46 SS17
47 SS17
56 SS17
57 SS17
62 SS17
66 SS17
70 SS17
72 SS17
104 SS17
110 SS17
115 SS17
116 SS17
122 SS17
87 SS17
88 SS17
90 SS17
91 SS17
147 SS20
148 SS20
150 SS20
152 SS20
153 SS20
155 SS20
160 SS20
162 SS20
177 SS21
178 SS34
179 SS34
180 SS34
183 SS34
44 SS17
165 SS20
143 SS20
156 SS20
107 SS17
86 SS17
96 SS17
168 SS28
94 SS17
35 SS17
112 SS17
118 SS17
Date
18-Jun
21-Jun
18-Jun
18-Jun
18-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
21-Jun
21-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
17-Jun
18-Jun
18-Jun
18-Jun
18-Jun
18-Jun
21-Jun
21-Jun
21-Jun
21-Jun
18-Jun
18-Jun
18-Jun
18-Jun
18-Jun
18-Jun
21-Jun
21-Jun
17-Jun
21-Jun
21-Jun
21-Jun
21-Jun
17-Jun
21-Jun
18-Jun
18-Jun
18-Jun
17-Jun
21-Jun
21-Jun
21-Jun
17-Jun
18-Jun
18-Jun
Stage
BR
BP
BR
GS
BR
BR
BR
BR
BR
BP
BR
BR
BP
BR
BR
BR
BR
BR
BR
BR
BR
BR
BR
BR
BR
BR
BR
BR
BR
BP
BR
GS
BR
BR
BR
BP
BR
BP
BR
BR
BP
BR
BR
BR
BR
BP
BR
BR
BP
BR
BR
BR
BR
BR
BR
BR
BP
GS
BR
BR
BP
BR
Species
Culcita novaeguineae
Culcita novaeguineae
Linckia guildingi
Linckia laevigata
Mithrodia clavigera
Mithrodia clavigera
Mithrodia clavigera
Mithrodia clavigera
Mithrodia clavigera
Mithrodia sp.
Mithrodia sp.
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
Most similar species
Culcita novaeguineae
Culcita novaeguineae
Linckia guildingi
Linckia laevigata
Mithrodia clavigera
Mithrodia clavigera
Mithrodia clavigera
Mithrodia clavigera
Mithrodia clavigera
Mithrodia bradleyi
Mithrodia clavigera
Arbacia lixula
Asterinides sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Choriaster sp.
Echinometra sp.
Eleutherozoa sp.
Heliocidaris erythrogramma
Liza argentea
Meridiastra calcar
Neoferdina cumingi
Ophionereis vittata
Oreaster reticulatus
Temnopleurus reevesii
Zoroaster ophiactis
Zoroaster ophiactis
Zoroaster ophiactis
CO1 Query cover(%) Ident(%)
100
100
96
96
85
98
85
100
91
99
81
99
90
99
84
98
98
97
56
92
85
91
13
91
84
82
97
83
93
80
93
78
95
83
95
82
94
80
98
82
93
80
94
82
90
81
99
82
49
81
50
81
95
79
89
82
80
81
89
81
60
81
87
81
98
82
72
80
99
82
84
80
99
82
45
82
95
82
93
81
95
82
84
80
96
81
98
83
88
81
88
78
88
82
65
81
90
82
68
82
99
81
99
97
97
76
40
80
57
82
73
83
95
82
90
81
96
79
96
80
80
78
93
80
Diversity 2016, 8, 9; doi:10.3390/d8020009
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(a) Temperature
(b) Salinity
Diversity 2016, 8, 9; doi:10.3390/d8020009
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(c) NH4+
(d) NO2−
Diversity 2016, 8, 9; doi:10.3390/d8020009
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(e) NO3−
(f) PO43−
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(g) SiO2
(h) DO
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(i) Turbidity
Figure S1. Distributions of temperature (a); salinity (b); nutrient concentrations (c–g); NH4+, NO2−,
NO3−, PO43−, and SiO2); (h) dissolved oxygen (DO); and (i) turbidity at the surface and at a depth of 7
m in Sekisei Lagoon during 3 days of sampling.
References
1.
Arndt, A.; Marquez, C.; Lambert, P.; Smith, M.J. Molecular phylogeny of Eastern Pacific sea cucumbers
(Echinodermata: Holothuroidea) based on mitochondrial DNA sequence. Mol. Phylogenet. Evol. 1996, 6,
425–437.