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SPECIES DIVERSITY AND PHYLOGENETIC RELATIONSHIPS OF SYMBIOTIC

CRUSTACEANS ON PORTUNUS PELAGICUS (LINNAEUS, 1758) IN VIETNAM

Conference Paper · November 2018

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 SPECIES DIVERSITY AND PHYLOGENETIC RELATIONSHIPS OF
SYMBIOTIC CRUSTACEANS ON PORTUNUS PELAGICUS
(LINNAEUS, 1758) IN VIETNAM
1
BINH T. DANG, 2OANH TK LE, 3SANG. Q. TRAN, 4OANH. TT, 5HENRIK GLENNER
1,3,4
Institute for Biotechnology and Environment, Nha Trang University, 02 Nguyen Dinh Chieu, Nha Trang City, Vietnam
2
Vietnam – Russia Tropical Center, 30 Nguyen Thien Thuat, Nha Trang City, Vietnam
5
Department of Biological Science, University of Bergen Postboks 7803, N-5020 Bergen
E-mail: [email protected]

Abstract - Blue swimming crab (Portunus pelagicus) was the economic and ecological important species in Vietnam, and
potential subject for aquaculture as well. More than 400 crab individuals were collected along the Vietnamese coastline, and
examined for ectosymbiotic crustacean. High diversity of 7 species have been reported, and 7 probably new for science. The
significant differences were detected between the infestation status of common occurring symbionts against the crab host
distribution (north, cemter, and south), sexes, and size groups. Symbiotic crustacean occupied specific niche(s) on crab body.
Molecular phylogeny supported the separation of 2 distinct lineages represented for two subclass (Thecostraca and
Copepoda. Further studies need to be conducted for new species description.

Keywords - Symbionts, Crustacean, Portunus pelagicus, Phylogeny, Infection, Vietnam

I. INTRODUCTION (Klinbunga et al., 2007), (Babu et al., 2012),

(Machado et al., 2013). Several species of symbiosis
The swimming crab, Portunus pelagicus is a major crustaceans has been reported to have impact on host
economical species throughout Indo-Pacific to the population function and dynamic (Babu et al., 2012),
coast of Africa (Galil and Innocenti, 1999). In (Hudson and Lester, 1994), (Isaeva et al., 2005). In
Vietnam, it is distributed in the wild throughout the Vietnam, 13 species of symbionts and parasites have
long coastal waters and islands (Ha et al., 2014) so far been identified. Six (three symbiosis
Throughout its native range, it is a valued marketable crustaceans) of those found were in both high
organism with numerous reports of it commercial prevalence and intensity. Of which, five species are
particulars from specific areas in its distribution, known to have negative impact on host populations
particularly, a multimillion dollar export commodity (Vo et al. 2013).
between Viet Nam and Japan and the U.S. (VASEF During some past decades, molecular markers were
Newsletter, Issue 21 November 5, 2013). Crabs play increasingly applied to investigate species diversity
an ecological important role through complex food and phylogenetic relationships. A number of research
web of coastal and marine ecosystems, especially in recently focused to examine phylogenetic position
mangrove forests, seagrass beds and coral reefs and evolution of cirripedia and their related barnacles
(Kunsook et al., 2014a), (Kunsook et al., 2014b). species (Wu, 2011), (Filipiak et al., 2016),
Additionally, their carbonate shells are widely known (Kwiatkowski et al., 2012), (Pe et al., 2018), (Mizrahi
as the living substrates of many epibont/symbionts et al., 1998), (Yusa et al., 2012). So far, no research
organisms (Abelló and Corbera, 1996), (Machado et has focused on clarifying the phylogenetics
al., 2013), (Machado et al., 2013), (Gaddes and relationships of the symbiosis crustacean community
Sumpton, 2004), (Dvoretsky, 2012). on blue swimming crab.
The symbiotic relationship, referred to permanent or The current research aims to conduct first
long-term association between two or more different comprehensive study for species diversity, infestation
species, is well known to be quite common in marine status, and molecular phylogeny of symbiosis
ecosystems. Reciprocal selection pressure between crustacean associations occurring on P. pelagucus
two interacting species can potentially altering the distributed along Vietnamese coastline to investigate
diversity, function and community dynamics of their the impact on host function and population structure.
hosts (Galil and Innocenti 1999). Data on the
symbionts community of Portunus pelagicus was II. MATERIAL AND METHODS
diversity depend on geographic location (Shields,
1992), (Shields and Wood, 1993). Among the 2.1. Symbiosis crustacean sampling and infestation
symbiont crustacean species, the barnacles frequently status
found attached to decapod crustaceans received the Blue swimming crab (Portunus pelagicus) was
most attention. Their symbiotic association depends collected across Vietnamese coastline from Cat Ba
on host’s biological characteristics such as Island - Hai Phong, Ha Long Bay - Quang Ninh in the
distribution, sex, maturity stage, molt cycle and size North; Nha Trang and Van Phong Bay - Khanh Hoa,
(Weng, 1987), (Gaddes and Sumpton, 2004), Song Cau and Tuy Hoa - Phu Yen in the Center, and

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 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam

Phu Quoc Island and Rach Gia - Kien Giang in he PCR reactions were performed using a total volume
South. The crabs were transported alive in aerated sea of 50 µl with components as following concentration:
water to the laboratory where they were kept in 10 µL of DreamTaq (Thermo Fisher Scientific) buffer
aquaria until dissected. 10X, 2 µL dNTP (10 mM), 2 µL each primer (10
For each crab, the sex, size (carapace width (CW) and mM), 1.25 unit of DreamTaq polymerase (5U/µl), 5
body weight (BW)) were recorded. Carapace width of µl DNA template and distilled water to the final
80 mm roughly corresponds to the size-at-maturity volume. Amplification was implemented using the
for P. pelagicus in Vietnam (Ha et al., 2014). To following PCR profile: a preliminary denaturation at
examine size differences in the levels of infestation, 94 °C for 3 minutes (min), followed by 35 cycles of
the crabs were subdivided into four groups: CW = < 94 °C for 45 seconds (s), annealing for 45 s (for 28S
80, 80 – 99, 100 – 119, and > 120 mm. rDNA, CO1 mtDNA gene at 56 °C, 42 °C,
Crabs were examined externally for symbiosis respectively), and then 72 °C for 45 s. This was
crustaceans. The carapace and abdomen of each crab followed by a final extension period at 72 °C for 7
was lifted off the body for dissection. For infected min before the samples were cooled to 4 °C. PCR
sites examination, the crab individuals were divided products were run on 1.5% agarose gel for
into 5 separate parts: (1) Carapace, (2) Limb confirmation of equal length against an appropriate
(maxilliped, chelliped, and swimming legs, (3) size markers. The PCR products were purified using
Abdomen, including egg clutches (if any), (4) Mouth DNA purification kits (Promega) and pre-sequenced
part, and (5) Gill. Each part was separated into petri using dye– labels dideoxy terminator (Big Dye
dishes containing clean seawater for inspection by the Terminator 3.1, Applied Biosystems) with the same
naked eye and stereoscope (Olimpus SZX9). primer as the PCR reactions at the following
Symbiosis crustacean species intended as whole- temperatures: 96 °C for 30 s, 50 °C for 30 s and 60
mounts were transferred to a vial of 70% alcohol, and °C for 4 min. Sequences of both strands were
those for DNA analysis were preserved in 95% generated on an ABI PRISM 3100 Genetic Analyzer
EtOH, and stored at -20°C. Symbiosis crustaceans (Applied Biosystems) using the amplification
were identified using various keys and species primers.
description following (Shields, 1992), (Shields and Sequence contigs were assembled using the Geneious
Wood, 1993) (Shields and Overstreet, 2003), (Jeffries Pro 5.5.7 (Kearse et al., 2012). The resulting
et al., 2005). sequences were confirmed by the Basic Local
We test the difference between crab distribution Alignment Search Tool (BLAST,
(geographical locations as north, center, and south), https://blast.ncbi.nlm.nih.gov/Blast.cgi). The
crab sex (male and female), and size group (4 groups) sequences were aligned, analyzed using BioEdit
against prevalence and mean intensity (defined in 7.0.5.3 (Hall, 1999) and submitted to GenBank.
(Margolis et al., 1982) and (Rózsa et al., 2000) of Information on genes application, and Genbank
each found symbiosis crustacean species using a one- accession numbers were presented in Table 1.
way ANOVA performed with Tukey’s post hoc tests. Phylogenetic trees were constructed using obtained
Data were tested for nominal distribution, and those 28S rDNA and COI mtDNA of collected symbiotic
of non-nominal distribution were log-transformed to crustaceans. Xia test was implemented for each
reduce differences in the variances between groups. sequence set in DAMBE 6.4.101 (Xia, 2013) to check
All statistical analyses were performed using using for substitution oversaturation based on the concept
SPSS free trial ver. 24.0 (IBM Corp. Released 2016. of entropy information theory. According to
IBM SPSS Statistics for Windows, Version 24.0. (Gontcharov et al., 2004), combined gene analysis
Armonk, NY: IBM Corp.). enhanced phylogenetic resolution, 2 current
sequence sets (7 sequences from each gene) were
2.2. Phylogenetic relationships combined with 17 available Genbank sequences
DNA was extracted from individual of each collected (Table 1, lacking genes marked as x) using Geneious
symbiont species using DNAeasy Tissue Extraction Pro 5.5.7 (Kearse et al., 2012). An incongruence-
Kit (Qiagen) in accordance with the manufacturer’s length difference (ILD) test (Barker and Lutzoni,
instructions. 28S rDNA and COI (Cytochrome c 2002) was conducted in PAUP*4.0b10 (Swofford,
oxidase subunit I) mitochondrial DNA were 2002) with 1000 randomized replicates to estimate
amplified using primer LSU 5 and 1500R (Olson et any difference in phylogenetic signal among the
al., 2003), and LCO1490 and HCO2198 (Folmer et different molecular sections.
al., 1994), respectively.

Table 1: Species list, sampling localities, Genbank accession numbers for all sequences of symbiotic crustaceans used in the
phylogenetic analysis (*sequences from Genbank. x sequences absent, W waiting Accession number)
Subclass Supperorder Order/Suboder Family Species COI mtDNA 28S rRNA

Lepadiformes/ Octolasmis angulata MH753551 MH727736

Thecostraca Thoracica Poecilasmatidae
Lepadomorpha Octolasmis neptuni W MH727737

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 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam

Octolasmis warwickii MH753552 MH727739

Octolasmis cor* KC138499 EU082326

Octolasmis hawaiense* KF484230 x

Octolasmis sp.* x EU082327

Temnaspis tridens MH753553 MH727738

Chelonibia cf. testudinaria MH753554 W

Chelonibia testudinaria* KJ754819 AB723914

Chelonibiidae
Chelonibia patula* JF823730 EU082295

Chelonibia caretta* JN589810 KM217526

Sessilia/ Coronulidae Xenobalanus globicipitis* KM217560 AB723927

Balanomorpha
Semibalanus cariosus* KM611728 AY520593
Archaeobalanidae
Semibalanus balanoides* MF748337 EU370440

Balanomorpha sp. MH753555 MH727740

Tetraclitella purpurascens* x AY520604

Tetraclitidae
Tetraclitella divisa* x AY520603

Sacculina carcini* KT209175 x

Rhizocephala Kentrogonida Sacculinidae
Sacculina confragosa* AB197803 x

Nicothoidae Choniosphaera indica W W

Siphonostomatoida Asterocheridae Asterocheres lilljeborgi* KR049050 KR048868

Copepoda Podoplea Lernaeopodidae Parabrachiella hugu* KT030285 KR048861

Canthocamptus staphylinus* MF077881 MF077853

Harpacticoida Canthocamptidae
Canthocamptus coreensis* KT030277 KR048886

Phylogenetic trees were constructed using 3 number of trees to be discarded in the ‘‘burn in’’ and
approaches, i.e., Maximum likelihood (ML), a final 50% majority-rule consensus
Maximum parsimony (MP) and Bayesian inference tree was constructed from the remaining trees.
(BI). Prior to ML and BI analyses, best-fit models of Numbers at the interior branches of the majority-rule
nucleotide substitution were selected by the Akaike consensus tree represent posterior probability (PP).
Information Criterion as implemented by Modeltest
3.7 (Posada and Crandall, 1998) and MrModeltest 2.2 III. RESULTS AND DISCUSSION
(Nylander, 2004). ML analyses were conducted using
the MEGA 6 (Tamura et al., 2013). The support of 3.1. Crustacean symbionts diversity and infection
clades was tested by the bootstrap method containing In total, 7 symbiotic crustaceans (Table 2, Fig. 1)
1000 replicates. MP analyses were conducted using were found occupying P. pelagicus distributed along
PAUP*4.0b10 (Swofford, 2002) . Trees were found Vietnamese coastline. Among that, Balanomorpha
by 1,000 replicate heuristic searches using the tree- sp., and Chelonibia cf. testudinaria were probably
bisection reconnection (TBR) branch-swapping new for science. The most common crustacean
algorithm, with 10 starting trees obtained by stepwise symbionts was the peduncle barnacle Octolasmis
addition. BI analyses were conducted in MrBayes angulata (Prevalence 75.57%, Mean intensity
3.2.6 (Huelsenbeck and Ronquist, 2001) 57.26±93.06), Its two congeners (O. neptuni and O.
under the selected best-fit models and parameters. warwickii), closed related species Temnaspis
Four chains were used and the analysis was run for tridens (Young, 2001) and an acorn barnacle
1 million generations, with the sampling frequency of Chelonibia cf. testudinaria were found to have
100. Each analysis was repeated twice to check moderate abundance (>10%), while Balanomorpha
for similarity of the likelihood plateau. Additionally, sp., and egg eating copepod (Choniosphaera indica)
parameter values were evaluated for occurred with low prevalence (0.21%) (Table 2).
convergence throughout the run by using the ‘‘sump’’
command in MrBayes and by examining Octolasmis spp. showed the variety of preferred
results in Tracer 1.3 (Drummond et al., 2005). Plots infected sites. O. warwickii infected most of the body
from Tracer were used to determine the appropriate parts of the crabs, except for the gills, while its two
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 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam

congeners do not found appear on the carapace. T. groups as only 2 species found on small crab (<80
tridens was found in most of body parts, and absent mm), 6 in middle sizes, and 7 species in large crabs
from the carapace. (Table 2, Fig. 2). Chelonibia cf. (>120 mm) (Table 3).
testudinaria occupied the outer surface (carapace and
limb), and C. indica exclusively on the abdomen of Five of seven species showed the significant
female crab (Table 2, Fig. 2). difference in prevalence and/or mean intensity among
Geographically, 6 species were discovered on P. host crab distribution (north, center, and south).
pelagicus in each sampling sites with Chelonibia cf. Levels of infestation was also depended on crab
testudinari occurring both in the north and south, sexes. Two Octolasmis and 1 Temnaspis species
while Balanomorpha sp. specify in the north and presented the significantly distinction in the
center. Five species (4 stallked and 1 acorn barnacle infestation rate. Prevalence and mean intensity
species) are commonly detected in both sexes, increased with crab size. Of which, 2 species showed
Balanomorpha sp. and C. indica found only in female significant different in 4 size groups, and 5 species in
crabs. Species numbers increased with crab size three size groups (Table 3).

Table 2: Infestation status of symbiosis crustacean species found on the blue swimming crabs, Portunus pelagicus (N= 479) along
Vietnamese coastline. Infection sites were marked as (1) Carapace, (2) Limb, (3) Abdomen, (4) Mouth part, and (5) Gill. Prevalences
and mean intensities (±SD) following P. pelagicus distribution (north, center, and south) and sex (male and female). Different letters
(in bold) mean prevalence and mean intensity was significantly different (one way ANOVA; P < 0.05).
Prevalence (P) Crab populations (P/MI) Crab sex (P/MI)
Infection
Name of species Mean Intensity
sites North Center South Male Female
MI)
Octolasmis 75.57 (2), (3), 82.55 72.00 73.08 77.12 74.07
angulata 57.26±93.06 (5) 65.63y±73.2 63.47y±16.75 37.01x±71.47 35.23x±46.46 79.53y±19.37
11.48 (2), (3), 9.00b 27.69c 13.98 9.05
O. neptuni 0.67a/2.00x
21.55±30.26 (5) 6.61x±5.56 29.56y±35.1 15.48±17.80 30.64±40.94
b b a a
15.45 (1), (2), 16.78 25.5 1.54 9.75 20.99b
O. warwickii x y x
23±31.12 (3), (4) 4.92 ±7.94 33.40 ±34.92 4.5 ±4.95 17.65±22.47 25.41±4.04
Temnaspis 19.00 (2), (3), 26.85b 21.00b 6.92a 15.25a 22.63b
tridens 7.95±8.49 (4), (5) 8.18±7.82 6.26±5.17 14.78±17.58 5.61±4.87 9.47±9.89
Chelonibia cf. 14.82 45.64b 2.31a 13.98 15.64
(1)
testudinari 9.32±9.96 9.68y± 0.3 1.33x±0.58 10.03±0.26 8.71±9.64
Balanomorpha 4.03
0.21*/6 (3) 0.50/6.00 0.41/6.00
sp. 18.67±16.90
Choniosphaera 2.09 1.50 0.77 4.12
(1)
indica 20.90±4.97 19.33±7.36 39.00 20.9±14.94
Species number 7 6 6 6 5 7

3.2. Phylogenetic relationships Octolasmis species were clustered together. T. tridens

In total, 7 sequences were generated from each gene was sister group with Octolasmis species, supporting
region of 7 species belonging to 4 families of separating into Temnaspis, Balanomorpha sp. was
crustacean symbionts on P. pelagicus distributing placed as sister species to Semibalanus species. Its
along Vietnamese coastline. The aligned data taxonomic position has not been clearly defined,
contained unambiguous 598 bp and 921 bp of COI although both morphological and genetic
mtDNA and 28S rRNA genes, respectively. The two characteristics have been investigated. Chelonbia cf.
sequence sets passed Xia test (lss < lss.c, in which lss testudinari was closed to C, caratta, and they both
values were significantly lower (P < 0.05) than lss.c, sister clade to C. testudinaria and previously
indicating that there was no substantial substitution conspecific C. patuda. The second lineage (subclass
saturation. The concatenated sequences yielded 606 Podoplea) consists of two sister clades (represented
(39.9%) constant characters and 748 (46.2%) for two orders Harpacticoida and
parsimony-informative characters out of 1519 Siphonostomatoida). The nicothoid copepode
characters. ILD test (PAUP 4.0b10) supported (Choniosphaera indica) was placed as separated
combination data sets (P < 0.05) (Cunningham, clade (Figure 3).
1997). The best selected model was GTR+I+G. High diversity of symbiotic crustaceans have been
Different analysis methods performed similar firstly detected on P. pelagicus in Vietnam. With
topologies (Figure 3), except minor different in MP intensive examination (over 400 crab individuals),
phylogenetic tree. With combined 28S+COI data set, rizocephala Saculina has not been found, similar to
two monophylectic lineages were detected previously reported by Vo et al. (2013).
represented for two subclass (Thecostraca and Balanomorpha sp. was morphological similar to
Copepoda). Semibalanus species, however, molecular phylogeny
The first lineage included species belong to two showed it appears as closed but different taxon. We
supperoders: Thoracica included two orders also found Chelonibia cf. testudinari, its
(Lepadiformes and Sesilia), and Rhizocephala (order morphological characters is similar to C.
Kentrogonida containing Saculina species). All testudinaria/C. pandula, however, molecular

Proceedings of ISER 169th International Conference, Bangkok, Thailand, 21st-22nd November, 2018
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 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam

phylogeny turned out to be sister to C. carella. parasite/symbiont differences between sexed mostly
Sequence differences compared to C. testudinaria/C. due to low growth rate (in female), distinct feeding
pandula (4.4 and 5.6% respectively), as well as C. and migration as well as Sacullina infection. Molting
carella (3.6%) (data not showed) can be seen as behavior was also the vital factor relating to the
three distinct species. abundant of symbiotic species. Additionally,
Considering the host features (distribution, sexes, and Seasonal fluctuations in species composition are also
size group), we found the significant differences in frequently observed in blue swimming crabs ((Jeffries
most common occurring symbionts. This has been et al., 2005). However, small differences in
documented in previous studies on various geographic distribution (such as case studies at the
crustaceans (Dvoretsky, 2012), (Shields and coast of Vietnam) are rarely detected.
Overstreet, 2003), (Dvoretsky and Dvoretsky, 2010), Infection sites of symbiotic crustaceans was also
(Ekanem et al., 2013) and swimmer crabs (Weng, recorded in the same way as previous studies
1987. Shield (1992)). Shield (1992) explained the ((Gaddes and Sumpton, 2004; Yusa et al., 2012)

Table 3: Prevalences and mean intensities (±SD) of symbiosis crustaceans following P. pelagicus size groups, Different letters (in
bold) mean prevalence and mean intensity was significantly different (one way ANOVA; P < 0.05).

Crab size group (Prevalence/Mean Intensity)

Name of species
<80 80-99 100-119 >120
23.53a/13.69x±36.3 72.73b/19.44x±26. 85.04c 96.92d/103.29z±129.
Octolasmis angulata
4 19 /49.23y±66.1 2
O. neptuni 0 3.25a/6.20±5.15 11.81b/7.67±5.56 26.92c/29.69±35.02
17.32b/20.73±24.0
O. warwickii 0 9.47a/15.20±24.48 28.46c/27.51±35.98
3
Temnaspis tridens 0 11.04a/4.06x±2.75 20.47b/5.42x±4.76 36.92c/10.69y±10.26
Chelonibia cf.
4.41a/2.67±1.70 13.64b/7.57±9.28 9.45b/7.38±6.71 26.92c/11.46±11.09
testudinari
Balanomorpha sp. 0.77/6
Choniosphaera
1.95/ 9.33±6.02 1.57/ 21.00±11.00 3.85/27.80±15.82
indica
Species number 2 6 6 7

Fig. 1. Line drawing of symbiotic crustaceans on P. pelagicus A Balanomorpha sp , B. Chelonibia cf. testudinaria , C. Choniosphaera
indica, D. Temnaspis tridens, E Octolasmis angulata , F. O. neptuni, G. O. warwickii

Following Spear et al (1994), Thecostraca, represented by the subclasses Ascothoracida and Cirripedia (the
latter including the Acrothoracica, Rhizocephala, and Thoracica is mornophyletic taxon. Here we detected
Thecostraca monophyly: Thoracica (Lepadiformes/Sesilia)/Rhizocephala (Kentrogonida). The sister relationship
of Thoracica and Rizocephala has been previously reported (Hoeg et al, 2009), and also support here in.

CONCLUSIONS

We found seven species of crustacean on Portunus pelagicus. Among that, 2 probably new for science.
Common occurring species (5/7 species) show significant differences in distribution (north, central, south along

Proceedings of ISER 169th International Conference, Bangkok, Thailand, 21st-22nd November, 2018
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 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam

the coast of Vietnam), sexes and size groups of the host crab species. The phylogenetic tree supported two
separate lineages following the existing taxonomic system o crustacean species.

Fig. 2. Photo image description of infected sites of symbiosis crustaceans on Portunus pelagicus. A&B. Ventral and dorsal view of
heavy infected crab. C. Infected sites of C. cf. testudinaria (1), E. O. warwackii (2), G. Balanomorpha sp (3), D. O. angulata (4), F. O.
neptuni (5), H. T. tridens (6), and I. C. indica (7)

Fig.3. Phylogenetic tree of symbiotic crustacean species resulting from the Bayesian inference analysis of combined 28S rRNA and
COI mtDNA dataset. BT and PP values were presented on the branch.

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 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam

ACKNOWLEDGMENTS Biol. Evol. 21, 612–624.

https://doi.org/10.1093/molbev/msh052
[14] Ha, V.V., Nhan, T.H., Cuong, T. Van, Doan, N.S., 2014.
This project was funded by the NORAD through Stock and fishery assessment report of blue swimming crab
Norhed Project QZA-0485 SRV-13/0010 Portunus pelagicus (Linnaeus, 1758) in Kien Giang waters,
titled Incorporating Climate Change into Ecosystem Viet Nam.
[15] Hall, T.A., 1999. BioEdit: a user-friendly biological sequence
Approaches to Fisheries and Aquaculture
alignment editor and analysis program for Windows
Management in Sri Lanka and Vietnam. We thank for 95/98/NT. Nucleic Acids Symp. Ser. https://doi.org/citeulike-
team members of Biodiversity and Conservation, article-id:691774
Institute for Biotechnology and Environment, Nha [16] Hudson, D.A., Lester, R.J.G., 1994. Parasites and symbionts
Trang University for sampling support. of wild mud crabs Scylla serrata (Forskal) of potential
significance in aquaculture. Aquaculture 120, 183–199.
https://doi.org/10.1016/0044-8486(94)90077-9
REFERENCES [17] Huelsenbeck, J.P., Ronquist, F., 2001. MRBAYES: Bayesian
inference of phylogenetic trees. Bioinformatics 17, 754–755.
[1] Abelló, P., Corbera, J., 1996. Epibiont bryozoan (Bryozoa, https://doi.org/10.1093/bioinformatics/17.8.754
Ctenostomatida) of the crab Goneplax rhomboides [18] Isaeva, V. V., Dolganov, S.M., Shukalyuk, A.I., 2005.
(Brachyura, Goneplacidae) off the Ebro delta (western Rhizocephalan barnacles - Parasites of commercially
Mediterranean). Miscel-lània Zoològica 19, 43–52. important crabs and other decapods. Russ. J. Mar. Biol. 31,
[2] Babu, M.Y., Durgekar, R., Devi, V.J., Ramakritinan, C.M., 215–220. https://doi.org/10.1007/s11179-005-0075-9
Kumaraguru, A.K., 2012. Influence of ciriped barnacles [19] Jeffries, W.B., Voris, H.K., Naiyanetr, P., Panha, S., 2005.
Chelonibia patula ( Ranzani ) on commercial crabs from Gulf Pedunculate Barnacles of the Symbiotic Genus Octolasmis
of Mannar and Palk bay coastal waters. Res. Environ. Life (Cirripedia: Thoracica: Poecilasmatidae) from the Northern
Sci. 5, 109–116. Gulf of Thailand. Nat. Hist. J. Chulalongkorn Univ. 5, 9–13.
[3] Barker, F.K., Lutzoni, F.M., 2002. The utility of the [20] Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung,
incongruence length difference test. Syst. Biol. 51, 625–637. M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S.,
https://doi.org/10.1080/10635150290102302 Duran, C., Thierer, T., Ashton, B., Meintjes, P., Drummond,
[4] Cunningham, C.W., 1997. Can three incongruence tests A., 2012. Geneious Basic: An integrated and extendable
predict when data should be combined? Mol. Biol. Evol. 14, desktop software platform for the organization and analysis
733–740. of sequence data. Bioinformatics 28, 1647–1649.
https://doi.org/10.1093/oxfordjournals.molbev.a025813 https://doi.org/10.1093/bioinformatics/bts199
[5] Drummond, A.J., Rambaut, A., Shapiro, B., Pybus, O.G., [21] Klinbunga, S., Khetpu, Æ.K., Khamnamtong, B., Menasveta,
2005. Bayesian coalescent inference of past population Æ.P., 2007. Genetic Heterogeneity of the Blue Swimming
dynamics from molecular sequences. Mol. Biol. Evol. 22, Crab ( Portunus pelagicus ) in Thailand Determined.
1185–1192. https://doi.org/10.1093/molbev/msi103 Biochem. Genet. 725–736. https://doi.org/10.1007/s10528-
[6] Dvoretsky, A.G., 2012. Epibionts of the great spider crab, 007-9110-1
Hyas araneus (Linnaeus, 1758), in the Barents Sea. Polar [22] Kunsook, C., Gajaseni, N., Paphavasit, N., 2014a. The
Biol. 35, 625–631. https://doi.org/10.1007/s00300-011-1087- feeding ecology of the blue swimming crab, Portunus
x pelagicus (Linnaeus, 1758), at Kung Krabaen bay,
[7] Dvoretsky, A.G., Dvoretsky, V.G., 2010. Epifauna associated Chanthaburi Province, Thailand. Trop. Life Sci. Res. 25, 13–
with an introduced crab in the Barents Sea: A 5-year study. 27.
ICES J. Mar. Sci. 67, 204–214. [23] Kunsook, C., Gajaseni, N., Paphavasit, N., 2014b. A stock
https://doi.org/10.1093/icesjms/fsp243 assessment of the Blue Swimming crab Portunus Pelagicus
[8] Ekanem, A.P., Eyo, V.O., Ekpo, I.E., Bassey, B.O., Ekanem, (Linnaeus, 1758) for sustainable management in Kung
A.P., 2013. Parasites of Blue Crab ( Callinectes amnicola ) in Krabaen Bay, gulf of Thailand. Trop. Life Sci. Res. 25, 41–
the Cross River Estuary , Nigeria. Int. J. Fish. Aquat. Stud. 1, 59.
18–21. [24] Kwiatkowski, M., Engelstädter, J., Vorburger, C., 2012. On
[9] Filipiak, A., Zaj, K., Kübler, D., Kramarz, P., 2016. Genetic Specificity in Symbiont-Mediated Host-Parasite
Coevolution of cophylogeny associations and methods for Coevolution. PLoS Comput. Biol. 8.
studying their Abstract Coevolution may be defined as the https://doi.org/10.1371/journal.pcbi.1002633
process of reciprocal , adaptive genetic change in two or [25] Machado, G.B.O., Sanches, F.H.C., Fortuna, M.D., Costa,
more species . Host-parasite interactions play an important T.M., 2013. Epibiosis in decapod crustaceans by stalked
role in the evolutionar 56–65. barnacle Octolasmis lowei (Cirripedia: Poecilasmatidae).
[10] Folmer, O., BLACK, M., HOEH, W., Lutz, R., Vrijenhoek, Zool. 30, 307–311. https://doi.org/10.1590/S1984-
R., 1994. DNA primers for amplification of mitochondrial 46702013000300007
cytochrome c oxidase subunit I from diverse metazoan [26] Margolis, L., Esch, G.W., Holmes, J.C., Kuris, A.M., Schad,
invertebrates. Mol. Mar. Biol. Biotechnol. 3, 294–299. G.A., 1982. The Use of Ecological Terms in Parasitology
https://doi.org/10.1371/journal.pone.0013102 (Report of an Ad Hoc Committee of the American Society of
[11] Gaddes, S.W., Sumpton, W.D., 2004. Distribution of barnacle Parasitologists). J. Parasitol. 68, 131.
epizoites of the crab Portunus pelagicus in the Moreton Bay https://doi.org/10.2307/3281335
region, eastern Australia. Mar. Freshw. Res. 55, 241–248. [27] Mizrahi, I., Achituv, Y., Katcoff, D.J., Perl Treves, R., 1998.
https://doi.org/10.1071/MF02136 Phylogenetic position of Ibla (Cirripedia: Thoracica) based
[12] Galil, B.S., Innocenti, G., 1999. NOTES ON THE on 18S rDNA sequence analysis. J. Crustac. Biol. 18, 363–
POPULATION STRUCTURE OF THE PORTUNID CRAB 368.
CHARYBDIS LONGICOLLIS LEENE , PARASITIZED [28] Nylander, J.A.A., 2004. MrModeltest v2. Program distributed
BY THE RHIZOCEPHALAN HETEROSACCUS by the author. Evol. Biol. Cent. Uppsala Univ. 2, 1–2.
DOLLFUSI BOSCHMA , OFF THE MEDITERRANEAN [29] Olson, P.D., Cribb, T.H., Tkach, V. V., Bray, R.A.,
COAST OF ISRAEL The defining feature of the fauna off Littlewood, D.T.J., 2003. Phylogeny and classification of the
the Mediterranean coast of Is 64, 451–463. Digenea (Platyhelminthes: Trematoda). Int. J. Parasitol. 33,
[13] Gontcharov, A.A., Marin, B., Melkonian, M., 2004. Are 733–755. https://doi.org/10.1016/S0020-7519(03)00049-3
Combined Analyses Better Than Single Gene Phylogenies? A [30] Pe, M., Høeg, J.T., Kolbasov, G.A., Crandall, K.A., 2018.
Case Study Using SSU rDNA and rbcL Sequence REANALYSIS OF THE RELATIONSHIPS AMONG THE
Comparisons in the Zygnematophyceae (Streptophyta). Mol. CIRRIPEDIA AND THE ASCOTHORACIDA AND THE
PHYLOGENETIC POSITION OF THE FACETOTECTA (

Proceedings of ISER 169th International Conference, Bangkok, Thailand, 21st-22nd November, 2018
37
 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam
MAXILLOPODA : THECOSTRACA ) USING 18S rDNA version 6.0. Mol. Biol. Evol. 30, 2725–2729.
SEQUENCES In the most recent phylogenetic studies of the https://doi.org/10.1093/molbev/mst197
Crustacea Maxillopoda , membe 22, 661–669. [38] Vo T.D, Bristow, G.A, Pham N.H, N.T.H.T., 2013. Some
[31] Posada, D., Crandall, K.A., 1998. MODELTEST: Testing the parasites found from swimming crab (Porttunus pelagicus
model of DNA substitution. Bioinformatics 14, 817–818. Linaeus, 1766) caught in Khanh Hoa marine water. J. Fish.
https://doi.org/10.1093/bioinformatics/14.9.817 Sci. Technol. Nha Trang Univ. 11–15.
[32] Rózsa, L., Reiczigel, J., Majoros, G., 2000. Quantifying [39] Weng, H.., 1987. Parstitic barnacle, Sacculina granifera
parasites in samples of hosts. J. Parasitol. 86, 228–232. Boschma, affecting the commercial sand crab Portunus
https://doi.org/10.1645/0022- pelagicus (L.), in populations from two diffrent environments
3395(2000)086[0228:QPISOH]2.0.CO;2 in Queensland. J. Fish Dis. 221–227.
[33] Shields, J.D., 1992. PARASITES AND SYMBIONTS OF [40] WU, T.H., 2011. Phylogeny and Population Genetics of
THE CRAB POR TUNUS PELAGICUS FROM MORETON Acorn Barnacles in Family Chthamalidae (Crustacea:
BAY, EASTERN AUSTRALIA. Crustac. Biol. 12, 94–100. Cirripedia). PhD thesis.
[34] Shields, J.D., Overstreet, R.M., 2003. The Blue Crab: [41] Xia, X., 2013. DAMBE5: A comprehensive software package
Diseases, Parasites, and Other Symbionts. Blue Crab Dis. for data analysis in molecular biology and evolution. Mol.
Parasites Other Symbionts 119. Biol. Evol. 30, 1720–1728.
[35] Shields, J.D., Wood, F.E.I., 1993. Impact of parasites on the https://doi.org/10.1093/molbev/mst064
reproduction and fecundity of the blue sand crab Portunus [42] Young, P.S., 2001. Deep-sea Cirripedia Thoracica
pelagicus from Moreton Bay, Australia. Mar. Ecol. Prog. Ser. (Crustacea) from the northeastern Atlantic collected by
92, 159–170. https://doi.org/10.3354/meps092159 French expeditions. Zoosystema 23, 705–756.
[36] Swofford, D.L., 2002. PAUP*. Phylogenetic Analysis Using [43] Yusa, Y., Yoshikawa, M., Kitaura, J., Kawane, M., Ozaki, Y.,
Parsimony and Other Methods). Version 4.0b10. . .x. Sinauer Yamato, S., Høeg, J.T., 2012. Adaptive evolution of sexual
Assoc. https://doi.org/Doi: 10.1111/j.0014- systems in pedunculate barnacles. Proc. R. Soc. B Biol. Sci.
3820.2002.tb00191 279, 959–966. https://doi.org/10.1098/rspb.2011.1554
[37] Tamura, K., Stecher, G., Peterson, D., Filipski, A., Kumar, S.,
2013. MEGA6: Molecular evolutionary genetics analysis

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