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Supplemen al ma e ial
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Seek and you shall ind: De ec ion o alien b yozoans along he Chilean SE Paci ic coas wi h
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a simple and cos -e icien me hodology
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Running i le: De ec ion o alien b yozoans along he Chilean SE Paci ic coas
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Sabine Rech1,2,3*, Leand o M. Viei a4,5, And ea I. Va ela3,6, Ma ía Soledad Rome o Bas ías2,7,
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Juan Pablo Fuen es8, Dennis P. Go don9, Ma ina Cacciu olo-Said2, Ku Paschke10, Ja ie
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Sellanes2,3, Ma in Thiel2,3,11
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1 MARE - Ma ine and En i onmen al Sciences Cen e, Regional Agency o he De elopmen
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o Resea ch, Technology and Inno a ion (ARDITI), Edi . Madei a Tecnopolo, Caminho da
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Pen eada, Piso 2, 9020-105 Funchal, Po ugal
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2 Depa amen o de Biologia Ma ina, Facul ad de Ciencias del Ma , Uni e sidad Ca ólica del
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No e, Coquimbo, Chile
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3 Cen e o Ecology and Sus ainable Managemen o Oceanic Islands (ESMOI), Facul ad de
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Ciencias del Ma , Uni e sidad Ca ólica del No e, La ondo 1281, Coquimbo, Chile
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4 Labo a ó io de Es udos de B yozoa (LAEB y), Depa amen o de Zoologia, Cen o de
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Biociências, Uni e sidade Fede al de Pe nambuco, A . P o . Mo aes Rego 1235, Cidade
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Uni e si á ia, 50670-901 Reci e, PE, B azil
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5 Depa men o Sciences, Na u al His o y Museum, C omwell Road, London SW7 5BD, UK
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6 Depa amen o de Oceanog a ía, Facul ad de Ciencias Na u ales y Oceanog á icas,
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Uni e sidad de Concepción, Ba io Uni e si a io S/N, Concepción, Chile
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7 Labo a o io de Mic oscopía Elec ónica, Dp o. de Biología Ma ina, Uni e sidad Ca ólica del
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No e, La ondo 1281, Coquimbo, Chile
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8 Es ación Expe imen al Quempillén, Ins i u o de Ciencias Ma inas y Limnológicas,
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Uni e sidad Aus al de Chile, Valdi ia, Chile
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9 New Zealand Ins i u e o Ea h Science (NZIES), P i a e Bag 14901, Kilbi nie, Welling on
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6241, New Zealand
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10 Ins i u o de Acuicul u a, Uni e sidad Aus al de Chile, Los Pinos S/N, Pelluco, Pue o Mon ,
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Chile
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11 Ma ineGEO, Smi hsonian En i onmen al Resea ch Cen e , 647 Con ees Wha Road,
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Edgewa e , MD 21037, USA
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* = co esponding au ho
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Table S1. S udy loca ions and da es. * = Resul s om Rapa Nui we e published in Rech e
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al. (2024a). S = aus al sp ing/ summe , W = aus al au umn/ win e . ** = in local in e idal.
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*** = on plas ic li e . ID = Si e iden i ica ion code. La = La i ude, Long = Longi ude.
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Si e
ID
La
Long
Time pe iod
(dd.mm.yyyy)
Si e use
S udy si es (se lemen pla es)
Rapa Nui, Hanga Piko*
RN
27.15°S
109.43°W
S: 20.11.2022 - 16.03.2023
Ma ina
Coquimbo,
Uni e si y campus
COQ
29.97°S
71.35°W
W: 22.02.2022 – 22.06.2022
S: 28.09.2022 – 07.02.2023
Aquacul u e/
Po
Alga obo,
Co adía Náu ica del Pací ico
ALG
33.36°S
71.69°W
W: 07.03.2022 - 11.07.2022
S: 21.09.2022 - 31.01.2023
Ma ina
San An onio,
In e na ional Po
SAN
33.58°S
71.62°W
W: 18.05.2022 - 18.09.2022
Po
Pue o Mon ,
Yach ing Club Relonca i
PMO
41.49°S
72.99°W
W: 13.05.2022 - 13.09.2022
S: 13.09.2022 - 23.01.2023
Ma ina
Ancud,
Resea ch s a ion Quempillen
ANC
41.87°S
73.78°W
W: 11.05.2022 - 15.09.2022
S: 16.09.2022 - 24.01.2023
Aquacul u e
Addi ional si es (oppo unis ic and beach su ey)
Coquimbo,
Uni e si y campus
COQ
29.97°S
-71.35°W
02.09.2023**
Aquacul u e/
Po
Ri oque, Beach
RIT
32.82°S
-71.52°W
22.04.2021***
Beach
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Table S2. Specimens deposi ed in he biological collec ions SCBUCN (Sala de colecciones
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biológicas, Uni e sidad Ca ólica del No e). COQ = Coquimbo, SAN = San An onio, PMO
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= Pue o Mon , ANC = Ancud, RIT = Ri oque, RN = Rapa Nui. * = Resul s om Rapa Nui
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we e published in Rech e al. 2024a.
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Species
Subs a um
Si e
Museum code
Bugulina s oloni e a
(Ryland, 1960)
Se lemen pla es (sea su ace)
COQ
SCBUCN-8176,
SCBUCN -8177
SAN
SCBUCN-8178
PMO
SCBUCN-8179
Bugulina c . ul a
(Ryland, 1960)
Se lemen pla es (sea su ace)
COQ
SCBUCN-8180
Conopeum sp.
Se lemen pla es (sea su ace)
ANC
SCBUCN-8182
Elec a monos achys
(Busk, 1854)
Plas ic deb is (beach)
RIT
SCBUCN-10540,
SCBUCN -10541
Wa e sipo a a cua a
(Ban a, 1969)
Rocks (in e idal)
COQ
SCBUCN-8181,
SCBUCN -10539
Wa e sipo a sub o qua a
(d’O bigny, 1852)
Se lemen pla es (sea su ace)
RN
SCBUCN-5570*
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4
Table S3. Resul s o Pe mu a ional ANOVA (PERMANOVA), showing deg ees o eedom
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(d ), sums o squa es (SS), mean squa es (M'), pseudo-F a io (Pseudo-F) and pe mu a ion P-
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alue (P(pe m)) and numbe o unique pe mu a ions (Unique pe ms) o each ac o . Sums o
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squa es ype: Type III (pa ial). Fixed e ec s sum o ze o o mixed e ms. Pe mu a ion me hod:
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Pe mu a ion o esiduals unde a educed model. Numbe o pe mu a ions: 9999. Please see
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Table S1 o ull si e names. *Te m has one o mo e emp y cells. Bold w i ing = s a is ically
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signi ican alues.
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A e age species ichness pe sampling uni (= ame)
Based on Euclidean dis ance. All si es and seasons included.
Main es
Sou ce
d
SS
MS
Pseudo-F
P(pe m)
Unique
pe ms
Si e
5
62.889
12.578
113.2
0.0001
9960
Season
1
4
4
36
0.0003
286
Si exSeason*
3
13.5
4.5
40.5
0.0001
9073
Residuals
9
1
0.11111
To al
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78.105
Communi y composi ion pe sampling uni (= ame)
B ay-Cu is simila i y. All si es and seasons. excep Ancud (win e ).
Main es
Sou ce
d
SS
MS
Pseudo-F
P(pe m)
Unique
pe ms
Si e
5
33228
6645.6
110.75
0.0001
9921
Season
1
234.59
234.59
3.9094
0.1204
9984
Si exSeason*
2
1000.9
500.45
8.34
0.0109
9972
Residuals
8
480.05
60.006
To al
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37025
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Table S4. F equency o occu ence, indica ed as he pe cen age o se lemen pla es
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ouled, o each axon o each combina ion o si e and season. Visualized by a colou scale,
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anging om 0% (ligh yellow) o 100% (da k g een). win = aus al au umn/win e , sum =
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aus al sp ing/summe . Ø, S . De = A e age equency and s anda d de ia ion o each species
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amongs all combina ions o si e and season. n (se lemen pla es) = 4 o San An onio (win e );
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n = 8 o all o he combina ions o si e and season. NIS = non-indigenous species, c yp =
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c yp ogenic, na = na i e.
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Table S5. Measu emen s o Bugulina s oloni e a. COQ = Coquimbo. SAN = San An onio.
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PMO = Pue o Mon . SCBUCN = Biological collec ion o Uni e sidad Ca ólica del No e,
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Coquimbo, Chile. BMNH = Na u al His o y Museum, London, UK. MMF = Museu de His ó ia
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Na u al do Funchal, Madei a. SERC = Smi hsonian En i onmen al Resea ch Cen e specimen
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lib a y, Edgewa e , Ma yland, USA. x = in o ma ion no gi en in sou ce publica ion. n =
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numbe o examined uni s o each ea u e.
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ou s udy
Ryland
1960
Ramalhosa
e al. 2017
McCann
e al. 2019
Loca ion
Chile,
COQ
Chile,
SAN
Chile,
PMO
Uni ed
Kingdom
Madei a
Galapagos
Museum
ID
SCBUCN
8176
SCBUCN
8177
SCBUCN
8178
SCBUCN
8179
BMNH
1959.3.18.1-
1959.3.18.4
MMF
44803-
44807
SERC
232836;
310273
Zooid
leng h,
ZL [µm]
Min –
Max
458 – 898
430 – 774
514 –1078
471 – 855
x
461–675
x
Mean
± SD
637 ± 110
594 ± 85
761 ± 151
665 ± 93
700 ± 75 -
740 ± 43
595
560 ± 64
n
35
29
24
35
x
x
x
Zooid
wid h,
ZW [µm]
Min –
Max
105 – 178
119 – 157
130 –182
112 –168
x
100 – 165
x
Mean
± SD
140 ± 15
137 ± 10
160 ± 14
143 ± 14
165 ± 13 -
180 ± 15
133
145 ± 17
n
34
24
22
36
x
x
x
ZL / ZW
Mean
± SD
4.6 ± 0.9
4.3 ± 0.7
4.6 ± 0.9
4.7 ± 0.7
4.2 ± 0.4 -
4.3 ± 0.6
x
x
n
34
23
20
33
x
x
x
A icula ia
leng h,
AL [µm]
Min –
Max
94 – 206
109 –209
123 – 217
82 – 241
x
97 – 155
x
Mean
± SD
164 ± 30
173 ± 27
174 ± 32
171 ± 42
x
140
165 ± 19
n
24
20
14
29
x
x
x
A icula ia
wid h,
AW [µm]
Min –
Max
72 – 144
64 – 132
76 – 131
42 – 134
x
x
x
Mean
± SD
109 ± 19
104 ± 17
102 ± 14
95 ± 25
x
x
137 ± 8
n
25
22
14
29
x
x
x
7
AL / AW
Mean
± SD
1.5 ± 0.2
1.7 ± 0.2
1.7 ± 0.3
1.9 ± 0.4
x
x
x
n
23
20
11
25
x
x
x
O icell
leng h,
OL [µm]
Min –
Max
103 – 144
94 – 139
100 – 134
122 – 157
x
101 - 124
x
Mean
± SD
125 ± 13
124 ± 11
116 ± 11
136 ± 9
x
122
156 ± 4
n
9
27
13
19
x
x
x
O icell
wid h,
OW [µm]
Min –
Max
162 – 219
154 – 194
153 -208
167 – 211
x
140 - 164
x
Mean
± SD
191 ± 17
177 ± 9
186 ± 18
195 ± 10
x
157
164 ± 9
n
9
26
13
19
x
x
x
OL / OW
Mean
± SD
0.7 ± 0.1
0.7 ± 0.1
0.6 ± 0.1
0.7 ± 0.0
x
x
x
n
9
26
13
17
x
x
x
65
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Table S6. Cy och ome c oxidase subuni I (COI) ba codes gene a ed in he p esen
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s udy. SCBUCN = Sala de colecciones biológicas de Uni e sidad Ca ólica del No e. GB =
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Genbank.
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Bugulina s oloni e a (SCBUCN #8176; GB #PP109395)
>B i4_consensus_F ame1
ACTTTATATTTTTTATTTGGCCTATGAGCTGGTATAGTTGGAAGAGGATTAAGTG
CCTTAATTCGAGTAGAACTTAGTCAACCAGGAAGTTTACTAGGTAATGACCACCT
TTATAACGTGATTGTTACAGCTCATGCCTTTTTAATAATTTTCTTTATAGTTATAC
CCGTAATAATTGGAGGATTTGGAAATTGATTAGTCCCTCTAATACTAGGATGCCC
AGATATAGCTTTTCCACGACTAAATAATATAAGATTTTGACTTTTACCACCTGCA
CTAGCTCTATTACTAATATCTTCTTTAGTTGAAAGAGGAGCAGGTACAGGATGAA
CAGTATACCCCCCTTTATCATCTAGTCTCGGTCATAGTGGACCATCAGTTGATTTA
GCAATTTTCTCCTTACACTTAGCTGGTGTCTCATCAATTTTAGGAGCAATTAATTT
TATAACATCAACAATGAATATACGAAGAGGATCAATAACTATAATACATATCCC
TCTATTAGTTTGAGCAGTTTTCATTACTGCTGTGTTACTACTACTATCTCTTCCAG
TTCTAGCAGGCGCAATTACTATGCTGTTAACAGATCGAAATCTTAACACATCTTT
TTTTGATCCTGC
Bugulina s oloni e a (SCBUCN #8177; GB #PP109396)
>B i13_consensus_F ame1
AAAGATATTGGAACTTTATATTTTTTATTTGGCCTTTGAGCTGGTATAGTTGGAA
GAGGATTAAGTGCCTTAATTCGAGTAGAACTTAGTCAACCAGGAAGTTTACTAG
GTAATGACCACCTTTATAACGTGATTGTTACAGCTCATGCCTTTTTAATAATTTTC
TTTATAGTTATACCCGTAATAATTGGAGGATTTGGAAATTGATTAGTCCCTCTAA
TACTAGGATGCCCAGATATAGCTTTTCCACGACTAAATAATATAAGATTTTGACT
TTTACCACCTGCACTAGCTCTATTACTAATATCTTCTTTAGTTGAAAGAGGAGCA
GGTACAGGATGAACAGTATACCCCCCTTTATCATCTAGTCTCGGTCATAGTGGAC
CATCAGTTGATTTAGCAATTTTCTCCTTACACTTAGCTGGTGTCTCATCAATTTTA
GGAGCAATTAATTTTATAACATCAACAATGAATATACGAAGAGGATCAATAACT
ATAATACATATCCCTCTATTAGTTTGAGCAGTTTTCATTACTGCTGTGTTACTACT
ACTATCTCTTCCAGTTCTAGCAGGCGCAATTACTATGCTGTTAACAGATCGAAAT
CTTAACACATCTTTTTTTGATCCTGCAGGAGGAGGAGATCCCATTTTATATCAAC
ATCTCTTCTGATTTTTTGGTCACC
Wa e sipo a a cua a (SCBUCN #8181; GB #PP109397)
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>B i6_FINAL2_F ame_1
GTAGTAACAGCTCACGCATTTTTCATGATTTTTTTTATAGTAATACCAATCATAAT
TGGAGGATTTGGAAACTGGTTAGTTCCTCTTATAATGGGAATCCCGGATATAGCA
TTCCCACGAATAAATAACATAAGATTTTGATTACTACCCCCTGCAGCTATTTTAC
TAATAATATCAGCACTAGTAGAAAGAGGTGCAGGGACAGGGTGAACAGTATACC
CCCCTTTATCTTCAAACATTTCTCATAGAGGATCATCAGTAGATATAGCAATTTTC
GCTCTTCACTTAGCAGGAGTGTCTTCAATTTTAGGAGCAATTAACTTTATAGTAA
CAATTATCAATATACGAGGAAAAATAATAGCATTCATAAACTTACCATTATTCGT
ATGAGCCGTATTTATCACTGCCCTATTATTATTACTATCTCTCCCAGTTCTAGCAG
GAGCTATTACTATACTACTAACAGACCGTAATTTTAACTCATCATTCTTTGACCC
AGCT
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16
96
97
Figu e S1. Examples o ouled se lemen pla es a e e ie al. Le : Coquimbo, summe
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2023. Righ : Alga obo, win e 2022.
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100
101
Figu e S2. Species ichness pe sample uni (= ame) by s udy si e and season. n ( ames)
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= 1 o San An onio (win e ); n = 2 o all o he combina ions o si e and season.
103
104
17
105
Figu e S3. nMDS (non-me ic mul idimensional scaling) plo o communi y composi ion
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based on he equency o occu ence o each axon on each sample uni a each combina ion
107
o s udy si e and season. win = win e , sum = summe . n (sampling uni s) = 1 o San An onio
108
(win e ); n = 2 o all o he combina ions o si e and season.
109
110
18
111
Figu e S4. Elec a monos achys, ound a ached o PET bo le agmen on Ri oque beach,
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on 22nd o Ap il 2021. Sample ID: SCBUCN #10540. a) colony o e iew. b) close up o
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au ozooids.
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19
115
Figu e S5. Wa e sipo a nig a, USNM 8495, holo ype, Galapagos. a) Zooids. b) O i ice
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(close-up). c) Colony o e iew.
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