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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: 1binhdt@ntu.edu.vn

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),

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

Proceedings of ISER 169th International Conference, Bangkok, Thailand, 21st-22nd November, 2018
31
 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
South. The crabs were transported alive in aerated sea
water to the laboratory where they were kept in
aquaria until dissected.
For each crab, the sex, size (carapace width (CW) and
body weight (BW)) were recorded. Carapace width of
80 mm roughly corresponds to the size-at-maturity
for P. pelagicus in Vietnam (Ha et al., 2014). To
examine size differences in the levels of infestation,
the crabs were subdivided into four groups: CW = <
80, 80 – 99, 100 – 119, and > 120 mm.
Crabs were examined externally for symbiosis
crustaceans. The carapace and abdomen of each crab
was lifted off the body for dissection. For infected
sites examination, the crab individuals were divided
into 5 separate parts: (1) Carapace, (2) Limb
(maxilliped, chelliped, and swimming legs, (3)
Abdomen, including egg clutches (if any), (4) Mouth
part, and (5) Gill. Each part was separated into petri
dishes containing clean seawater for inspection by the
naked eye and stereoscope (Olimpus SZX9).
Symbiosis crustacean species intended as whole-
mounts were transferred to a vial of 70% alcohol, and
those for DNA analysis were preserved in 95%
EtOH, and stored at -20°C. Symbiosis crustaceans
were identified using various keys and species
description following (Shields, 1992), (Shields and
Wood, 1993) (Shields and Overstreet, 2003), (Jeffries
et al., 2005).
We test the difference between crab distribution
(geographical locations as north, center, and south),
crab sex (male and female), and size group (4 groups)
against prevalence and mean intensity (defined in
(Margolis et al., 1982) and (Rózsa et al., 2000) of
each found symbiosis crustacean species using a one-
way ANOVA performed with Tukey’s post hoc tests.
Data were tested for nominal distribution, and those
of non-nominal distribution were log-transformed to
reduce differences in the variances between groups.
All statistical analyses were performed using using
SPSS free trial ver. 24.0 (IBM Corp. Released 2016.
IBM SPSS Statistics for Windows, Version 24.0.
Armonk, NY: IBM Corp.).
2.2. Phylogenetic relationships
DNA was extracted from individual of each collected
symbiont species using DNAeasy Tissue Extraction
Kit (Qiagen) in accordance with the manufacturer’s
instructions. 28S rDNA and COI (Cytochrome c
oxidase subunit I) mitochondrial DNA were
amplified using primer LSU 5 and 1500R (Olson et
al., 2003), and LCO1490 and HCO2198 (Folmer et
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
Lepadiformes/
Thecostraca Thoracica Poecilasmatidae
Lepadomorpha
Proceedings of ISER 169th International Conference, Bangkok, Thailand, 21st-22nd November, 2018
PCR reactions were performed using a total volume
of 50 µl with components as following concentration:
10 µL of DreamTaq (Thermo Fisher Scientific) buffer
10X, 2 µL dNTP (10 mM), 2 µL each primer (10
mM), 1.25 unit of DreamTaq polymerase (5U/µl), 5
µl DNA template and distilled water to the final
volume. Amplification was implemented using the
following PCR profile: a preliminary denaturation at
94 °C for 3 minutes (min), followed by 35 cycles of
94 °C for 45 seconds (s), annealing for 45 s (for 28S
rDNA, CO1 mtDNA gene at 56 °C, 42 °C,
respectively), and then 72 °C for 45 s. This was
followed by a final extension period at 72 °C for 7
min before the samples were cooled to 4 °C. PCR
products were run on 1.5% agarose gel for
confirmation of equal length against an appropriate
size markers. The PCR products were purified using
DNA purification kits (Promega) and pre-sequenced
using dye– labels dideoxy terminator (Big Dye
Terminator 3.1, Applied Biosystems) with the same
primer as the PCR reactions at the following
temperatures: 96 °C for 30 s, 50 °C for 30 s and 60
°C for 4 min. Sequences of both strands were
generated on an ABI PRISM 3100 Genetic Analyzer
(Applied Biosystems) using the amplification
primers.
Sequence contigs were assembled using the Geneious
Pro 5.5.7 (Kearse et al., 2012). The resulting
sequences were confirmed by the Basic Local
Alignment Search Tool (BLAST,
https://blast.ncbi.nlm.nih.gov/Blast.cgi). The
sequences were aligned, analyzed using BioEdit
7.0.5.3 (Hall, 1999) and submitted to GenBank.
Information on genes application, and Genbank
accession numbers were presented in Table 1.
Phylogenetic trees were constructed using obtained
28S rDNA and COI mtDNA of collected symbiotic
crustaceans. Xia test was implemented for each
sequence set in DAMBE 6.4.101 (Xia, 2013) to check
for substitution oversaturation based on the concept
of entropy information theory. According to
(Gontcharov et al., 2004), combined gene analysis
enhanced phylogenetic resolution, 2 current
sequence sets (7 sequences from each gene) were
combined with 17 available Genbank sequences
(Table 1, lacking genes marked as x) using Geneious
Pro 5.5.7 (Kearse et al., 2012). An incongruence-
length difference (ILD) test (Barker and Lutzoni,
2002) was conducted in PAUP*4.0b10 (Swofford,
2002) with 1000 randomized replicates to estimate
any difference in phylogenetic signal among the
different molecular sections.
Family Species COI mtDNA 28S rRNA
Octolasmis angulata MH753551 MH727736
Octolasmis neptuni W MH727737
32
 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
approaches, i.e., Maximum likelihood (ML),
Maximum parsimony (MP) and Bayesian inference
(BI). Prior to ML and BI analyses, best-fit models of
nucleotide substitution were selected by the Akaike
Information Criterion as implemented by Modeltest
3.7 (Posada and Crandall, 1998) and MrModeltest 2.2
(Nylander, 2004). ML analyses were conducted using
the MEGA 6 (Tamura et al., 2013). The support of
clades was tested by the bootstrap method containing
1000 replicates. MP analyses were conducted using
PAUP*4.0b10 (Swofford, 2002) . Trees were found
by 1,000 replicate heuristic searches using the tree-
bisection reconnection (TBR) branch-swapping
algorithm, with 10 starting trees obtained by stepwise
addition. BI analyses were conducted in MrBayes
3.2.6 (Huelsenbeck and Ronquist, 2001)
under the selected best-fit models and parameters.
Four chains were used and the analysis was run for
1 million generations, with the sampling frequency of
100. Each analysis was repeated twice to check
for similarity of the likelihood plateau. Additionally,
parameter values were evaluated for
convergence throughout the run by using the ‘‘sump’’
command in MrBayes and by examining
results in Tracer 1.3 (Drummond et al., 2005). Plots
from Tracer were used to determine the appropriate
Proceedings of ISER 169th International Conference, Bangkok, Thailand, 21st-22nd November, 2018
33
number of trees to be discarded in the ‘‘burn in’’ and
a final 50% majority-rule consensus
tree was constructed from the remaining trees.
Numbers at the interior branches of the majority-rule
consensus tree represent posterior probability (PP).
III. RESULTS AND DISCUSSION
3.1. Crustacean symbionts diversity and infection
In total, 7 symbiotic crustaceans (Table 2, Fig. 1)
were found occupying P. pelagicus distributed along
Vietnamese coastline. Among that, Balanomorpha
sp., and Chelonibia cf. testudinaria were probably
new for science. The most common crustacean
symbionts was the peduncle barnacle Octolasmis
angulata (Prevalence 75.57%, Mean intensity
57.26±93.06), Its two congeners (O. neptuni and O.
warwickii), closed related species Temnaspis
tridens (Young, 2001) and an acorn barnacle
Chelonibia cf. testudinaria were found to have
moderate abundance (>10%), while Balanomorpha
sp., and egg eating copepod (Choniosphaera indica)
occurred with low prevalence (0.21%) (Table 2).
Octolasmis spp. showed the variety of preferred
infected sites. O. warwickii infected most of the body
parts of the crabs, except for the gills, while its two
 Species Diversity and Phylogenetic Relationships of Symbiotic Crustaceans on Portunus Pelagicus (Linneus 1758) in Vietnam

congeners do not found appear on the carapace. T.
tridens was found in most of body parts, and absent
from the carapace. (Table 2, Fig. 2). Chelonibia cf.
testudinaria occupied the outer surface (carapace and
limb), and C. indica exclusively on the abdomen of
female crab (Table 2, Fig. 2).
Geographically, 6 species were discovered on P.
pelagicus in each sampling sites with Chelonibia cf.
testudinari occurring both in the north and south,
while Balanomorpha sp. specify in the north and
center. Five species (4 stallked and 1 acorn barnacle
species) are commonly detected in both sexes,
Balanomorpha sp. and C. indica found only in female
crabs. Species numbers increased with crab size
groups as only 2 species found on small crab (<80
mm), 6 in middle sizes, and 7 species in large crabs
(>120 mm) (Table 3).
Five of seven species showed the significant
difference in prevalence and/or mean intensity among
host crab distribution (north, center, and south).
Levels of infestation was also depended on crab
sexes. Two Octolasmis and 1 Temnaspis species
presented the significantly distinction in the
infestation rate. Prevalence and mean intensity
increased with crab size. Of which, 2 species showed
significant different in 4 size groups, and 5 species in
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
region of 7 species belonging to 4 families of
crustacean symbionts on P. pelagicus distributing
along Vietnamese coastline. The aligned data
contained unambiguous 598 bp and 921 bp of COI
mtDNA and 28S rRNA genes, respectively. The two
sequence sets passed Xia test (lss < lss.c, in which lss
values were significantly lower (P < 0.05) than lss.c,
indicating that there was no substantial substitution
saturation. The concatenated sequences yielded 606
(39.9%) constant characters and 748 (46.2%)
parsimony-informative characters out of 1519
characters. ILD test (PAUP 4.0b10) supported
combination data sets (P < 0.05) (Cunningham,
1997). The best selected model was GTR+I+G.
Different analysis methods performed similar
topologies (Figure 3), except minor different in MP
phylogenetic tree. With combined 28S+COI data set,
two monophylectic lineages were detected
represented for two subclass (Thecostraca and
Copepoda).
The first lineage included species belong to two
supperoders: Thoracica included two orders
(Lepadiformes and Sesilia), and Rhizocephala (order
Kentrogonida containing Saculina species). All
was sister group with Octolasmis species, supporting
separating into Temnaspis, Balanomorpha sp. was
placed as sister species to Semibalanus species. Its
taxonomic position has not been clearly defined,
although both morphological and genetic
characteristics have been investigated. Chelonbia cf.
testudinari was closed to C, caratta, and they both
sister clade to C. testudinaria and previously
conspecific C. patuda. The second lineage (subclass
Podoplea) consists of two sister clades (represented
for two orders Harpacticoida and
Siphonostomatoida). The nicothoid copepode
(Choniosphaera indica) was placed as separated
clade (Figure 3).
High diversity of symbiotic crustaceans have been
firstly detected on P. pelagicus in Vietnam. With
intensive examination (over 400 crab individuals),
rizocephala Saculina has not been found, similar to
previously reported by Vo et al. (2013).
Balanomorpha sp. was morphological similar to
Semibalanus species, however, molecular phylogeny
showed it appears as closed but different taxon. We
also found Chelonibia cf. testudinari, its
morphological characters is similar to C.
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.
Sequence differences compared to C. testudinaria/C.
pandula (4.4 and 5.6% respectively), as well as C.
carella (3.6%) (data not showed) can be seen as
three distinct species.
Considering the host features (distribution, sexes, and
size group), we found the significant differences in
most common occurring symbionts. This has been
documented in previous studies on various
crustaceans (Dvoretsky, 2012), (Shields and
Overstreet, 2003), (Dvoretsky and Dvoretsky, 2010),
(Ekanem et al., 2013) and swimmer crabs (Weng,
1987. Shield (1992)). Shield (1992) explained the
parasite/symbiont differences between sexed mostly
due to low growth rate (in female), distinct feeding
and migration as well as Sacullina infection. Molting
behavior was also the vital factor relating to the
abundant of symbiotic species. Additionally,
Seasonal fluctuations in species composition are also
frequently observed in blue swimming crabs ((Jeffries
et al., 2005). However, small differences in
geographic distribution (such as case studies at the
coast of Vietnam) are rarely detected.
Infection sites of symbiotic crustaceans was also
recorded in the same way as previous studies
((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.

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
ACKNOWLEDGMENTS
This project was funded by the NORAD through
Norhed Project QZA-0485 SRV-13/0010
titled Incorporating Climate Change into Ecosystem
Approaches to Fisheries and Aquaculture
Management in Sri Lanka and Vietnam. We thank for
team members of Biodiversity and Conservation,
Institute for Biotechnology and Environment, Nha
Trang University for sampling support.
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