Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations
Understanding of accurate phylogenetic relationship among Penaeidae shrimp is important for academic and fisheries industry. The Morphometric and Randomly Amplified Polymorphic DNA (RAPD) analysis was used to make the phylogenetic relationsip among 13 Penaeidae shrimp. For morphometric analysis fort...
Збережено в:
| Дата: | 2014 |
|---|---|
| Автори: | , , , |
| Формат: | Стаття |
| Мова: | English |
| Опубліковано: |
Інститут клітинної біології та генетичної інженерії НАН України
2014
|
| Назва видання: | Цитология и генетика |
| Теми: | |
| Онлайн доступ: | http://dspace.nbuv.gov.ua/handle/123456789/126670 |
| Теги: |
Додати тег
Немає тегів, Будьте першим, хто поставить тег для цього запису!
|
| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Цитувати: | Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations / P. Rajakumaran, B. Vaseeharan, R. Jayakumar, R. Chidambara // Цитология и генетика. — 2014. — Т. 48, № 6. — С. 17-24. — Бібліогр.: 52 назв. — англ. |
Репозитарії
Digital Library of Periodicals of National Academy of Sciences of Ukraine| id |
irk-123456789-126670 |
|---|---|
| record_format |
dspace |
| spelling |
irk-123456789-1266702017-12-02T03:03:19Z Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations Rajakumaran, P. Vaseeharan, B. Jayakumar, R. Chidambara, R. Оригинальные работы Understanding of accurate phylogenetic relationship among Penaeidae shrimp is important for academic and fisheries industry. The Morphometric and Randomly Amplified Polymorphic DNA (RAPD) analysis was used to make the phylogenetic relationsip among 13 Penaeidae shrimp. For morphometric analysis forty variables and total lengths of shrimp were measured for each species, and removed the effect of size variation. The size normalized values obtained was subjected to UPGMA (Unweighted Pair-Group Method with Arithmetic Mean) cluster analysis. For RAPD analysis, the four primers showed reliable differentiation between species, and used correlation coefficient between the DNA banding patterns of 13 Penaeidae species to construct UPGMA dendrogram. Phylogenetic relationship from morphometric and molecular analysis for Penaeidae species found to be congruent. We concluded that as the results from morphometry investigations concur with molecular one, phylogenetic relationship obtained for the studied Penaeidae are considered to be reliable. Понимание точных филогенетических отношений у креветок Penaeidae важно как с общенаучной точки зрения, так и для рыбной промышленности. RAPD анализ был использован для установления филогенетических связей 13 видов креветок Penaeidae. Для морфометрических анализов измерены 40 переменных и общих длин креветок для каждого вида и устранен эффект вариабельности размера. Показатели нормализованного размера обработаны с помощью кластерного анализа UPGMA (Unweighted Pair-Group Method with Arithmetic Mean). При RAPD анализе четыре праймера показали достоверные различия между видами. Коэффициенты корреляции между паттернами ДНК использованы для построения UPGMA дендрограмм. Филогенетические связи, построенные на основе морфометрических и молекулярных анализов, совпали, что позволило сделать вывод об их достоверности. 2014 Article Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations / P. Rajakumaran, B. Vaseeharan, R. Jayakumar, R. Chidambara // Цитология и генетика. — 2014. — Т. 48, № 6. — С. 17-24. — Бібліогр.: 52 назв. — англ. 0564-3783 DOI: 10.3103/S0095452714060103 http://dspace.nbuv.gov.ua/handle/123456789/126670 en Цитология и генетика Інститут клітинної біології та генетичної інженерії НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Оригинальные работы Оригинальные работы |
| spellingShingle |
Оригинальные работы Оригинальные работы Rajakumaran, P. Vaseeharan, B. Jayakumar, R. Chidambara, R. Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations Цитология и генетика |
| description |
Understanding of accurate phylogenetic relationship among Penaeidae shrimp is important for academic and fisheries industry. The Morphometric and Randomly Amplified Polymorphic DNA (RAPD) analysis was used to make the phylogenetic relationsip among 13 Penaeidae shrimp. For morphometric analysis forty variables and total lengths of shrimp were measured for each species, and removed the effect of size variation. The size normalized values obtained was subjected to UPGMA (Unweighted Pair-Group Method with Arithmetic Mean) cluster analysis. For RAPD analysis, the four primers showed reliable differentiation between species, and used correlation coefficient between the DNA banding patterns of 13 Penaeidae species to construct UPGMA dendrogram. Phylogenetic relationship from morphometric and molecular analysis for Penaeidae species found to be congruent. We concluded that as the results from morphometry investigations concur with molecular one, phylogenetic relationship obtained for the studied Penaeidae are considered to be reliable. |
| format |
Article |
| author |
Rajakumaran, P. Vaseeharan, B. Jayakumar, R. Chidambara, R. |
| author_facet |
Rajakumaran, P. Vaseeharan, B. Jayakumar, R. Chidambara, R. |
| author_sort |
Rajakumaran, P. |
| title |
Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations |
| title_short |
Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations |
| title_full |
Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations |
| title_fullStr |
Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations |
| title_full_unstemmed |
Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations |
| title_sort |
conformation of phylogenetic relationship of penaeidae shrimp based on morphometric and molecular investigations |
| publisher |
Інститут клітинної біології та генетичної інженерії НАН України |
| publishDate |
2014 |
| topic_facet |
Оригинальные работы |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/126670 |
| citation_txt |
Conformation of phylogenetic relationship of penaeidae shrimp based on Morphometric and Molecular investigations / P. Rajakumaran, B. Vaseeharan, R. Jayakumar, R. Chidambara // Цитология и генетика. — 2014. — Т. 48, № 6. — С. 17-24. — Бібліогр.: 52 назв. — англ. |
| series |
Цитология и генетика |
| work_keys_str_mv |
AT rajakumaranp conformationofphylogeneticrelationshipofpenaeidaeshrimpbasedonmorphometricandmolecularinvestigations AT vaseeharanb conformationofphylogeneticrelationshipofpenaeidaeshrimpbasedonmorphometricandmolecularinvestigations AT jayakumarr conformationofphylogeneticrelationshipofpenaeidaeshrimpbasedonmorphometricandmolecularinvestigations AT chidambarar conformationofphylogeneticrelationshipofpenaeidaeshrimpbasedonmorphometricandmolecularinvestigations |
| first_indexed |
2025-07-09T05:31:21Z |
| last_indexed |
2025-07-09T05:31:21Z |
| _version_ |
1837146134102933504 |
| fulltext |
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 6 17
Understanding of accurate phylogenetic relationship among
Penaeidae shrimp is important for academic and fisheries
industry. The Morphometric and Randomly amplified
polymorphic DNA (RAPD) analysis was used to make the
phylogenetic relationsip among 13 Penaeidae shrimp. For
morphometric analysis forty variables and total lengths of
shrimp were measured for each species, and removed the
effect of size variation. The size normalized values obtained
was subjected to UPGMA (Unweighted Pair-Group Method
with Arithmetic Mean) cluster analysis. For RAPD analysis,
the four primers showed reliable differentiation between
species, and used correlation coefficient between the DNA
banding patterns of 13 Penaeidae species to construct
UPGMA dendrogram. Phylogenetic relationship from
morphometric and molecular analysis for Penaeidae species
found to be congruent. We concluded that as the results
from morphometry investigations concur with molecular one,
phylogenetic relationship obtained for the studied Penaeidae
are considered to be reliable.
Key words: Penaeidae, shrimp, morphometry, RAPD, phylo-
geny.
Introduction. Penaeidae shrimps worldwide are
classified into 26 genera and 225 species, of which
13 genera are found along the Indian coasts. It
is a diverse and worldwide distributed family of
shrimps, particularly in the Indo-West Pacific re-
gion and contributes to 50 % of crustacean fisher-
ies in the world [1]. In India Penaeidae shrimps
constitute a highly valuable fishery along the East
and West coasts. The estimated Indian landings
of Penaeidae shrimps in 2010 was 2,17,900 mt,
contributing 51 % of total crustacean landings [2].
Generally, the Penaeidae shrimp differ from
non Penaeidae by pattern of arranging of pleurae
regularly, third pereopods chelated and abdominal
segment do not has sharp bend but non Penaeidae
prawns the pleurae of the second abdominal segment
are overlapping those of first and third segments;
third pereopods not chelated and abdominal seg-
ment has sharp bend in the non Penaeidae prawns.
Moreover in Penaeidae shrimp, the distinguished
identical characters are the rostral structure, rostral
teeth, antenna colouration and body colour with
strips. In majority of the Penaeidae shrimp, rostral
teeth are important characters to distinguish the dif-
ferent species and also within the groups. In Penaeus
sensu lato group, the rostral teeth are present both
in the upper and lower portions of the rostrum, and
but in other case, the rostral teeth are present only
in the dorsal side of the rostrum. Moreover depend-
ing on the nature of carapace the Penaeus sensu lato
can be divided as Melicertus (grooved) and non-
Melicertus (non-grooved). Carapace with longitu-
dinal suture, lacking transverse suture and fourth
and fifth pairs of pereopods with elongate dactyl
subdivided into articles in Xiphopenaeus, carapace
with longitudinal, transverse sutures, and fourth and
fifth pair of pereopods with dactyl neither elongate
nor subdivided into articles in Trachypenaeus.
Penaeidae into three tribes according to the
characters of gill formula (presence or absence of
epipod on the third maxilliped and pleurobranch
on the fifth pereopod), spination of the antennular
peduncle (presence or absence of a spine on the
ventromedian margin of proximal segment) and the
telson (tip simple or trifid), namely the Peneini
(Penaeus, Heteropenaeus, Funchalia, Pelagopena-
eus), the Parapeneini (Parapenaus, Artemesia, Pe-
naeopsis, Metapenaeopsis), and the Trachypeneini
(Metapenaeus, Macropetasma, Trachypenaeopsis, Aty-
popenaeus, Protrachypene, Xiphopenaeus, Parape-
naeopis, Trachypenaeus) [3]. Penaeidae shrimp, on
the basis of complex morphological similarity matrix
into five groups; 1) Penaeus; 2) Penaeopsis; 3) At-
popenaeus, Trachypenaeopsis, Metapenaeus; 4) Para-
penaeus, Parapenaeopsis, Trachypenaeus; 5) Metape-
naeopis [4].
The extraordinary morphological diversity among
these species poses substantial challenge to their
phylogenetic study. Limited fossil records and in-
© P. RAJAKUMARAN, B. VASEEHARAN,
R. JAYAKUMAR, R. CHIDAMBARA, 2014
CONFORMATION OF PHYLOGENETIC RELATIONSHIP OF PENAEIDAE SHRIMP
BASED ON MORPHOMETRIC AND MOLECULAR INVESTIGATIONS
P. RAJAKUMARAN 1, B. VASEEHARAN 1, R. JAYAKUMAR 2, R. CHIDAMBARA 3
1 Crustacean Molecular Biology and Genomics Lab, Department of Animal Health and Management, Alagappa University, India
2 Mandapam Regional Centre, Central Marine Fisheries Research Institute (CMFRI), Mandapam Camp – 623520, Tamil Nadu, India
3 R&D Department, National Prawn Co., Al-Laith 21961, Jeddah, Saudi Arabia
E-mail: vaseeharanb@gmail.com
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 618
P. Rajakumaran, B. Vaseeharan, R. Jayakumar, R. Chidambara
complete palaeogeographic evidences are of little
significant in phylogenetic studies of Penaeidae
[5, 6]. The system of morphometric measurements
called the truss network system have been widely
used for population and taxonomic studies [7–11].
The conservation of body skeleton and definite bo-
dy ratio facilitated using trussnet work system as a
taxonomical tool to differentiate physically similar
species and compared to traditional morphologi-
cal method such tool can be used as clear, ac-
curate and precise descriptor [12]. The morpho-
metric measurements have been used to describe
the taxonomical relationship between Melicertus ke-
rathurus, Metapenaeus dobsoni and Penaeus semisul-
catus [13]. Apart from morphological characters,
the shape and size evolutionary relationship species
of Penaeidae species is limited. Hence in the present
study the truss network system was used to analyses
the phylogenetic relationship by morphometry with
concerning shape and size.
The molecular analysis using PCR-based mar-
kers has been of great significance on studying the
phylogeny and taxonomy. Random amplification of
polymorphic DNA (RAPD) is random amplifica-
tion of anonymous loci by PCR. The method is
simple, rapid and cheap, has high polymorphism,
requires only a small amount of DNA and al-
lows creation of genomic markers from species of
which little information is known about the target
sequences to be amplified. RAPD markers are pro-
duced by PCR using short oligonucleotide primers
of random sequences. Different RAPD patterns
arise when genomic regions vary according to the
presence/absence of complementary primer anne-
aling sites. RAPDs have gained considerable at-
tention particularly in population genetics, species
and subspecies identification, phylogenetics, link-
age group identification, chromosome and genome
mapping, analysis of interspecific gene flow and
hybrid speciation, and analysis of mixed genome
samples breeding analysis and as a potential sour-
ce for single-locus genetic fingerprints [14]. Ran-
domly amplified polymorphic DNA (RAPD) data
showed higher levels of polymorphism than allo-
zymes [15]. Allozyme has limitations for highly
divergent group [15, 16].
The robust taxonomy is both the morphological
and molecular data agreement [17]. Molecular data
are a complementary approach to morphology, es-
pecially in discriminating cryptic or sibling species
[18, 19] and for constructing phylogenetic relation-
ships [15]. Raymunida – squat lobsters (formerly in
genus Munida), distinguished by small morphologi-
cal differences, which matched clear differences in
mitochondrial nucleotide sequences [20]. Data are
available from studies of mitochondrial marker like
Cytochrome oxidase subunit I, and 12S and 16S
rRNA (ribosomal RNA). However, analyses by
non nuclear marker alone or combined, still have not
provided a reliable taxonomy for Penaeidae shrimps.
Nuclear DNA marker only provides high resolution
to resolve the evolutionary relationship of the shrimp
[6, 21]. Only mitochondrial DNA marker observation
is available on Penaeidae phylogeny [22, 23], but lot
of studies on phylogeny of super family Penaeoidea
are there, where only the least explanation about the
Penaeidae shrimp as one of the family along with
four other family Aristaeidae, Solenoceridae, Benthe-
sicymidae, Sicyonidae [24–26]. Moreover the other
studies are Penaeus sensu lato [27, 28] and Penaeus
sensu stricto [29–31]. Previously several morphometric
and genetic analysis were conducted for species varia-
tion and systematic studies for various fishes [32–35].
Currently, there is no report available on phylogenetic
relationship of Penaeidae species analysed employ-
ing both morphometry and Nuclear DNA molecular
marker. In the present study, we have used both the
techniques, with diverse shrimp, to conform the phy-
logenetic relationship of Penaeidae shrimp species.
Materials and methods. Collection of samples.
Six species from Penaeus genus (1) Fenneropenaeus
indicus (H. Milne Edwards, 1837), (2) Fennero-
penaeus merguiensis (De Man, 1888), (3) Melicer-
tus latisulcatus (Kishinouye, 1896), (4) Penaeus
monodon (Fabricius, 1798), (5) Penaeus semisulca-
tus (De Haan, 1844), (6) Marsupenaeus japonicus
(Bate, 1888); three species from Parapenaeopsis
genus (1) Parapenaeopsis stylifera (H. Milne Ed-
wards, 1837), (2) Parapenaeopsis hardwickii (Miers,
1878), (3) Parapenaeopsis uncta (Alcock, 1905); two
species form Metapenaeus genus (1) Metapenaeus
monoceros (Fabricius, 1798), (2) Metapenaeus dob-
soni (Miers, 1878), Metapenaeus affinis (H. Milne
Edwards, 1837) and one species from Penaeopsis
genus Penaeopsis jerryi (Perez Farfante, 1979), to-
tally thirteen species were collected from the East
coast of India. All species were identified according
to published literature [36–38].
Multivariate cluster analysis of morphometric da-
ta. After identification of species, from each spe-
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 6 19
Conformation of phylogenetic relationship of Penaeidae shrimp based on morphometric
cies 5 male sample only taken and female omitted
due to size variation. The measurement were taken
on the basis of truss net work system (Fig. 1) in
which eighteen landmarks, forty factor or variables
plus total length of each shrimp (tip of rostrum to
end of telson), totally forty one variables. From the
five sample of each species the average value was
taken. The significant correlations were observed
between average morphometric measurement and
size of the species. Hence for removing the effect
of size variation for all species, the transforma-
tion of absolute measurement to size independent
variable were carried out, in which the formula
Madj = M(Ls/Lo)
b was used [39]. Where Madj the
size adjusted measurement of each variable, M is
the original measurement of each variable, Lo the
standard length of shrimp (total length from tip
of rostrum to end of telson) and Ls the overall
mean of measurement for all shrimp for each vari-
able. Allometric vs. standard estimates allometric
coefficients with respect to standard. The standard
variable was placed in the first column and each
additional column is regressed onto the first column
after log-transformation, giving a slope (allometric
coefficient) b for that variable. The Ls/Lo is com-
puted in the excel sheet and detecting the b param-
eter was carried out using paleontological statistics
software (PAST) package version 1.93. From these
sizes adjusted morphometric measurements for
each species were subjected to UPGMA multivari-
ate cluster analysis. Euclidean method (statistic tool
that quantifies the extent to which species within
clusters are similar to one another) was followed to
detect the morphometrical relationship of Penaei-
dae studied using paleontological statistics software
(PAST) package (Table 1–3 –http://cytgen.com/
articles/4860017s.pdf).
DNA isolation. Total genomic DNA was isolat-
ed from muscle tissue by SDS-phenol/chloroform
method described with slight modifications [40].
Briefly, shrimp muscle (200 mg) was cut into small
pieces, crushed using a sterile porcelain mortar and
pestle with 1 ml of chilled TEN buffer (50 mM Tris-
HCl, pH 8.0, 50 mM EDTA and 100 mM NaCl),
and transferred to 2 ml Eppendorf tube. Protein-
ase K 8 l (300 mg/ml), sucrose 20 l (2 %), and
20 l sodium dodecyl sulfate SDS (2 %) were added
to the tube. After overnight incubation at 60 ºC, the
lysate was extracted once with phenol and twice
with chloroform/isoamyl alcohol. DNA was pre-
cipitated with isopropanol, washed once with 70 %
ethyl alcohol, and suspended in TE (Tris EDTA,
pH 8.0) buffer. DNA quality and quantity were
determined by Agarose gel electrophoresis and Bio-
photometer plus («Eppendorf», Germany).
RAPD-PCR amplification and data analysis.
Eighteen primers were used for RAPD analysis.
DNA amplification reactions were performed in
200 mol/l each dNTP, 2 mmol/l MgCl2, 19 stan-
dard Taq polymerase buffer, 0.2 mol/l random
primer, 40 ng genomic DNA and 0.75 U Taq poly-
merase in a final volume of 25 l. PCR conditions
included initial denaturation at 94 ºC for 5 min,
followed by 45 cycles of denaturation at 94 ºC for
30 s, annealing at 35 ºC for 1 min, extension at 72 ºC
for 2 min and final extension at 72 ºC for 10 min.
The amplified DNA was separated by electropho-
resis through 2 % agarose gel containing Ethidium
bromide in 1×TBE buffer at a constant 80 V. To
maintain consistency, only the repeatable major
bands ranging from 10 000 to 1000 bp were scored.
Molecular weights of amplified bands were esti-
mated by comparing with known molecular weight
marker (1 Kbp DNA ladder, Bangalore Genie, In-
dia). DNA profiles generated for all samples were
compared in a pairwise manner. RAPD banding
patterns were recorded on spread sheets as binary
matrix marking alleles absent (0) and present (1).
The similarity index between species [41] and sub-
sequently the data used to construct a dendrogram
using the (UPGMA) algorithm, as described [42].
Results. Morphometrical relationship. When con-
verting the actual measurement value into size stan-
daisation value, there were lot of variation between
two kinds of value for most morphometrical vari-
Fig. 1. The truss network system
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 620
P. Rajakumaran, B. Vaseeharan, R. Jayakumar, R. Chidambara
able. The size adjusted value for all variables for all
species described the relationship of species. This
values were relatively closeness for Penaeus species,
Metapenaeus, Parapenaeopsis and Penaeopsis jerryi.
While these size adjusted morphometric value of
all variables subjected to UPGMA cluster analysis,
produced 10 clades. The first clade included P. jer-
ryi, M. latisulcatus second, M. japonicus third, P.
monodon – F. indicus fourth, F. merguiensis fifth,
P. semisulcatus sixth, M. affinis seventh, M. brevi-
cornis – M. dobsoni eighth, P. uncta ninth, and P.
stylifera – P. hardwickii tenth (Fig. 2).
Genetic relationship. For RAPD molecular analysis,
among 18 primers used, 10 primers responded, but only
four primer RM03 5 AAT CGG GCT G 3 , RM07 5
CAA TGC CCG T 3 , RM014 5 GTA TTG CCC T
3 , RM017 5 TCC CTC GTG C 3 generated good and
reproducible RAPD profiles for all the species stud-
ied. Totally 99 major DNA bands produced, 11 bands
polymorphic. The most number of banding pattern
were similar for F. indicus and P. monodon; P. semis-
ulcatus and F. merguiensis; M. latisulcatus and M. ja-
ponicus; P. stylifera, P. hardwickii and P. uncta. P. jerryi
banding pattern vary, however most of the bands close
to Metapenaeus and Parapenaeopsis species (Fig. 3).
Genetic similarity values range form 0–1 for all iso-
lates, and high similarity value for Penaeus genus
of non-Melicertus clade species of F. merguiensis P.
semisulcatus, F. indicus, P. monodon, than Melicertus
clade species of M. japonicus and M. latisulcatus.
As for Metapenaeus genus, the genetic similarity val-
ue for the species, M. affinis, M. dobsoni, M. bre-
vicornis were closely related, and Parapenaeopsis ge-
nus the species of P. stylifera, P. hardwickii, P. uncta
were coming closely. Penaeopsis species P. jerryi
genetic similarity value close to Metapenaeus and
the Parapenaeopsis. As construction of UPMGA
dendrogram based on genetic similarity, producing
9 clades, in which the order of relationship that
M. marsupenaeus – M. latisulcatus in first clade, P.
monodon – F. indicus in second, P. semisulcatus –
F. merguiensis in third, P. jerryi, in fourth, M. affinis in
fifth, M. monoceros – M. dobsoni sixth, P. uncta in sev-
enth, P. hardwickii – P. stylifera eighth in ninth (Fig. 4).
Fig. 2. Phylogeny of shrimps based on morphometric analysis using
PAST Package
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 6 21
Conformation of phylogenetic relationship of Penaeidae shrimp based on morphometric
Discussions. In the present study the compre-
hensive investigation of phylogenetic relationship
of Penaeidae species by increasing number of mor-
phometrical trait using truss net work system and
many primer for RAPD analysis, resulting con-
firmed the phylogenetic relationship of this family.
Penaeidae shrimps differ in variety of morphologi-
cal characteristics that are the expression of genetic
differences among them. There are, numerous stud-
ies of the morphological differences among spe-
cies which can be used for taxonomic distinctions.
Morphometric is a quantitative study of pattern of
covariance with shape [43] and many morphologi-
cal attributes of biological form are obviously re-
flection of evolutionary process [44–46], and since
assessment of pattern of morphologically variation
have been traditionally to infer phylogenetic rela-
tion [47–49] it would seem be natural to assume
that morphometric analyses would play a large role
in phylogenetic studies. The most of morphometri-
cal trait are unique for a species of Penaeidae in
our analysis, the closely related valued species more
phylogenetic related than other. Thus the morpho-
metric characters are describing the phylogenetic
relationship among tested penaeidae.
Though RAPDs are not sensitive to large-scale
length mutations, hence the variation might be un-
derestimated as the technique is based on the PCR
amplification of discrete regions of genome. The us-
ing of high number of primer in the present study,
among which the four producing good amplification
for all sample, hence the probability of reliability is
high in the present study. Detecting the phylogenetic
relatedness of species, on basis of DNA banding pat-
tern the among them. Thus both morphometry and
RAPD tool are helping to detect the phylogenetic
relationship of Penaeidae in the present study.
The phylogenetic relationship based on morpho-
logical traits such as grooved, non grooved on the
carapace, last abdominal somite, presence hepatic
ridge, shape of petasma, nature of thylecum open
or closed and coloration are used by [3, 4] and [37]
Fig. 3. RAPD profile of shrimps generated by polymerase chain reaction using four primer: M – Marker, Fi – F.
indicus, Pm – P. monodon, Ps – P. semisulcatus, Fm – F. merguiensis, Ml – M. latisulcatus, Mj – M. japonicus, Mb –
M. brevicornis, Md – M. dobsoni, Ma – M. affinis, Pas – P. stylifera, Pah – P. hardiwickii, Pau – P. uncta, Pj – P.
jerryi_Line shows similarity of DNA bands (Monomeric)
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 622
P. Rajakumaran, B. Vaseeharan, R. Jayakumar, R. Chidambara
for Penaeus s.l. not that much influence on evo-
lutionary relationship, because these morphological
characters could be diverged by sexual selection and
convergent evolution, plesiomorphic, so the Penae-
us s.l. group is to put old Penaeus genus, which is
both morphological and molecular data agreement
[32]. In the present study using both analyses, as
for Penaeus sensu lato, the Melicertus and Penae-
us sensu stricto (Non-Melicertus) are paraphyletic
which concurrent with previous phylogenetic stud-
ies [27, 28, 31, 50, 51]. The Fenneropenaeus and
Penaeus s.s. are sharing their relationship, since P.
monodon and P. semisulcatus combined with F. indi-
cus and F. merguensis these observation stand on the
view [22 28]. Thus Penaeus s.s., Fenneropanaeus,
Farfantepenaeus and Litopenaeus could be put on
another clade against Melicertus as non Melicertus.
In other Panaeidae genus such Metapenaeus and
Parapenaeopsis, which are coming as closely in both
analyses. This classification prevents the confusion
of academic and fishery industry.
New studies should be improved than prediction
of previous studies, in present study using four genera
of Penaeidae family described not only evolution-
ary relationship but also interspecies relationship at
genus level. For example as for Metapeneus species
M. dobsoni and M. brevicornis are closely related than
M. affinis. Parapenaeopsis species P. stylifera and P.
hardwickii are closely related than P. uncta. The P.
jerryi single species in Penaeopsis genus, which as
one of the sister genus in the Penaeidae family as
given by [3, 4] which form separate clade.
In the present study stand on [3] view that the
tribe Peneini diverged earlier than other two tribes,
with tribe Parapeneini and Trachypeneini sharing
common ancestor, Penaeidae form a monophyletic
group with a Penaeus shrimp as a common ancestor.
Our phylognetic observations also agree with find-
ings that evolutionary polarity of family was from
Penaeus to Trachypenaeus [52]. When combined
investigation of both morphometrical and genetical
(RAPD) analysis of Penaeidae species, the result that
both analysis outcomes are concurrent. The taxo-
nomical relationship of genus Penaeus, in which the
Melicertus and non Melicertus clade; Metapenaeus,
Parapenaeopsis and Penaeopsis are high agreement
in both analyses. Phylogenetic relationship devel-
oped through this study, not only helping conserva-
tion, systematic, ecological and evolutionary studies
would also help to produce superior captive shrimp
with economical trait strains through hybridization
of closely related species.
Conclusion. In the present study, phylogenetic
relationship of different genera Penaeus, Metape-
naeus, Parapenaeopsis, and Penaeopsis reported
based on molecular tool is congruent with mor-
phometric one. Thus it could be concluded that
morphometrical traits using truss net work system
more useful for phylogenic studies of Penaeidae
than traditional morphological and meristematic
traits. The comparative account of genetic (RAPD)
and morphometric based results would be a reliab-
le tool for confirmation of phylogenetic relation-
ship of these economically important species. Fur-
ther studies also needed to increase the numbers of
Penaeidae species.
I express my sincere gratitude to Department of Sci-
ence and Technology, New Delhi, India for the financial
assistance provided through DST-PURSE programme.
ÑÒÐÓÊÒÓÐÀ ÔÈËÎÃÅÍÅÒÈ×ÅÑÊÈÕ
ÑÂßÇÅÉ ÊÐÅÂÅÒÎÊ PENAEIDAE
ÍÀ ÎÑÍÎÂÅ ÌÎÐÔÎÌÅÒÐÈ×ÅÑÊÈÕ
È ÌÎËÅÊÓËßÐÍÛÕ ÈÑÑËÅÄÎÂÀÍÈÉ
P. Rajakumaran, B. Vaseeharan,
R. Jayakumar, R.Chidambara
Ïîíèìàíèå òî÷íûõ ôèëîãåíåòè÷åñêèõ îòíîøåíèé ó
êðåâåòîê Penaeidae âàæíî êàê ñ îáùåíàó÷íîé òî÷êè
çðåíèÿ, òàê è äëÿ ðûáíîé ïðîìûøëåííîñòè. RAPD
Fig. 4. Phylogenetic relatioship of Penaeidae shrimps as
per RAPD analysis using UPGMA algorithm
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 6 23
Conformation of phylogenetic relationship of Penaeidae shrimp based on morphometric
àíàëèç áûë èñïîëüçîâàí äëÿ óñòàíîâëåíèÿ ôèëî-
ãåíåòè÷åñêèõ ñâÿçåé 13 âèäîâ êðåâåòîê Penaeidae.
Äëÿ ìîðôîìåòðè÷åñêèõ àíàëèçîâ èçìåðåíû 40 ïå-
ðåìåííûõ è îáùèõ äëèí êðåâåòîê äëÿ êàæäîãî âèäà
è óñòðàíåí ýôôåêò âàðèàáåëüíîñòè ðàçìåðà. Ïîêà-
çàòåëè íîðìàëèçîâàííîãî ðàçìåðà îáðàáîòàíû ñ ïî-
ìîùüþ êëàñòåðíîãî àíàëèçà UPGMA (Unweighted
Pair-Group Method with Arithmetic Mean). Ïðè RAPD
àíàëèçå ÷åòûðå ïðàéìåðà ïîêàçàëè äîñòîâåðíûå ðàç-
ëè÷èÿ ìåæäó âèäàìè. Êîýôôèöèåíòû êîððåëÿöèè
ìåæäó ïàòòåðíàìè ÄÍÊ èñïîëüçîâàíû äëÿ ïîñòðîå-
íèÿ UPGMA äåíäðîãðàìì. Ôèëîãåíåòè÷åñêèå ñâÿçè,
ïîñòðîåííûå íà îñíîâå ìîðôîìåòðè÷åñêèõ è ìîëå-
êóëÿðíûõ àíàëèçîâ, ñîâïàëè, ÷òî ïîçâîëèëî ñäåëàòü
âûâîä îá èõ äîñòîâåðíîñòè.
REFERENCES
1. FAO Technical Guidelines for Responsible Fisheries.
Genetic Resource Management, Food and Agriculture
Organization. – Rome: FAO, 2008, ¹ 5. – P. 125.
2. CMFRI Annual Report 2010-2011. – Kochin, 2011. –
P. 163.
3. Burkenroad M.D. Natural classification of Dendro-
branchiata, with a Key to Recent Genera // Crus-
tacean Phylogeny / Ed. F.R. Schram. – Rotterdam,
1983. – P. 279–290.
4. Kubo I. Studies on the penaeids of Japanese and its
adjacent waters // J. Tokyo College Fish. – 1949. –
36, ¹ 1. – 467 p.
5. Perez-Farfante I. Western Atlantic shrimps of the
genus Penaeus // Fish. Bull. – 1969. – 67, ¹ 3. –
P. 461–590.
6. Dall W., Hill B.J., Rothlisberg P.C. Biology of the
Penaeidae // Advances in Marine Biology / Ed. D.J.
Sharples. – London : Acad. Press, 1990. – 27. – 489 p.
7. Strauss R.E., Bookstein F.L. The truss : Body form
reconstruction in morphometrics // Syst. Zool. –
1982. – 31, ¹ 1. – P. 113–135.
8. Lester L.J., Pante M.J.R. Genetic of Penaeus speci-
es // Marine shrimp culture : Principles and practi-
ces / Eds A.W. Fast, L.J. Lester. – Amsterdam :
Elsevier, 1992. – P. 29–52.
9. Lin Y.S., Tzeng C.S., Hwang J.K. Reassessment of
morphological characteristic in freshwater eels (ge-
nus Anguilla, Anguillidae) shows, congruence with
molecular phylogeny estimates // Zool. Scripta. –
2005. – 34, ¹ 3. – P. 225–234.
10. Kapiris K. Morphometric structure and allometry pro-
files of the giant red shrimp Aristaeomorpha foliacea
(Risso, 1827) in the Eastern Mediterranean //Natur.
History. – 2005. – 39, ¹ 17. – P. 1347–1357.
11. Turan C. Phylogenetic relationship of Mediterranean
Mullidae species (Perciformes inferred from genetic
and morphologic data // Sci. Mar. – 2006. – 70,
¹ 2. – P. 311–318.
12. Fitzgerald D.G., Nanson J.W., Todd T.N., Davis B.M.
Application of truss analysis for quantification of
changes in fish condition // Aquat. Ecosyst. Stress
Recov. – 2002. – 9. – P. 115–125
13. Aktas M., Cemal T., Bozkurt A. Taxonomic descrip-
tion of three shrimp species (Melicertus kerathurus,
Meatapenaeus monoceros, Penaeus semisulcatus) us-
ing multivariate morphometric analyses // Anim.Vet.
Adv. – 2006. – 5, ¹ 3. – P. 172–175.
14. Brown B., Epifanio J. Nuclear DNA // Population
Genetics : Principle and Application for Fisheries
Scientists / Ed. E.M. Hallerman. – Bethesda, 2003. –
P. 101–123.
15. Avise J.C. Molecular Markers, Natural History and
Evolution. – New York : Chapman and Hall, 1994. –
511 p.
16. Hillis D.M., Moritz C., Mable B.K. Application of
Molecular Systematics and the future of the field //
Molecular Systematics. – USA, 1996. – P. 514–543.
17. Dall W. Recent molecular research on Penaeus sensu
lato // Crustacean Biol. – 2006. – 27. – P. 380–
382.
18. Knowlton N. Sibling species in the seas // Annu .Rev.
Ecol. Syst. – 1993. – 24. – P. 189–216.
19. Sarver S.K., Silberman J.D., Walsh P.J. Mitochon-
drial DNA sequence evidence supporting the rec-
ognition of two subspecies or species of the Florida
spiny lobster Panilurus argus // Crustacean Biol. –
1998. – 18, ¹ 1. – P. 177–186.
20. Macpherson E., Machordom A. Phylogenetic relation-
ships of species of Raymunida (Decapoda: Galathei-
dae) based on Morphology and Mitochondrial cyto-
chrome oxidase sequences, with the recognition of
four new species // Crustacean Biol. – 2001. – 21,
¹ 3. – P. 696–714.
21. Vaseeharan B., Rajakamaran P., Jayaseelan D., Ye-
shvadha V.A. Molecular markers and their applica-
tion in genetic diversity of Penaeid shrimp // Aqua-
cult. Int. – 2013. – 21. – P. 219–241.
22. Chan T.Y., Tong J., Tam Y.K., Chu K.H. Phylogenet-
ic relationships among the genera of the Penaeidae
(Crustacea: Decapoda) revealed by Mitochondrial
16S rRNA gene sequences // Zootaxa. – 2008. –
1694. – P. 38–50.
23. Voloch C.M., Friere P.R., Russo C.A. Molecular phy-
logeny of penaeid shrimps inferred from two mito-
chondrial markers // Genet. Mol. Biol. – 2005. –
4, ¹ 4. – P. 668–674.
24. Quan J., Zhuang Z., Deng J. et al. Phylogenetic re-
lationship of 12 Penaeoidae shrimp species deduced
from Mitochondrial DNA sequences // Biochem.
Genet. – 2004. – 42, ¹ 9/10. – P. 331–345.
25. Vazquez-Bader A.R., Carrero J.C., Gárcia-Varela M.
et al. Molecular phylogeny of superfamily Penaeoi-
dea Rafinesque-Schmaltz, 1815, based on Mito-
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 624
P. Rajakumaran, B. Vaseeharan, R. Jayakumar, R. Chidambara
chondrial 16S partial sequence analysis // Shellfish
Res. – 2004. – 23, ¹ 3. – P. 911–917.
26. Ma K.Y., Chan T.Y., Chu K.H. Phylogeny of penae-
oid shrimps (Decapoda: Penaeoidea) inferred from
nuclear protein-coding genes // Mol. Phylogenet.
Evol. – 2009. – 53, ¹ 1. – P. 45–55.
27. Lavery S., TamY.K., Chu K.H. Phylogenetic relation-
ship and evolutionary history of the shrimp Penaeus
sp.l. derived from mitochondrial DNA // Mol. Phy-
logenet. Evol. – 2004. – 31, ¹ 1. – P. 39–49.
28. Ma K.Y., Chan T.Y., Chu K.H. Refuting the six-ge-
nus classification of Penaeus s.l. (Dendrobranchiata,
Penaeidae): a combined analysis of mitochondrial
and nuclear genes // Zool. Scripta. – 2011. – 40,
¹ 5. – P. 498–508.
29. Baldwin J.D., Bass A.L., Bowen B.W., Clark W.H.
Molecular phylogeny and biogeography of the ma-
rine shrimp Penaeus (Crustacea: Penaeidae) // Mol.
Phylogenet. Evol. – 1998. – 10, ¹ 3. – P. 399–407.
30. Gusmão S., Lazoski C., Solê-Cava A.M. A new spe-
cies of Penaeus revealed by allozyme and cytochro-
me oxidase I analysis // Marine Biol. – 2000. –
137, ¹ 3. – P. 435–446.
31. Wang Z.Y., Tsoi K.H., Chu K.H. Applications of
AFLP technology in genetic and phylogenetic anal-
ysis of penaeid shrimp // Biochem. Syst. Ecol. –
2004. – 32. – P. 399–440.
32. Benson McRae L.K. Morphometric and genetic con-
firmation of two species of Kuhlia (Osteichthyes :
Kuhliidae) in Hawai’i : Biology of Hawaiian streams
and Esturaries / Eds N.L. Evenhuis // Bishop Mu-
seum Bull. Cultur. Environ. – 2007. – 3. – P. 113–
123.
33. Dhinakaran A., Alikunhi N.M., Chinnathambi S. et al.
Assesment of Morphometric and Genetic variation in
three freshwater fish species of the genus Garra (Os-
teichthyes: Cyprinidae) // Not. Sci. Biol. – 2011. –
3, ¹ 1. – P. 12–26.
34. Young S., Ni M.H., MinY.L. Systematic study of
the Simocephalus Sensu Stricto species group (Cla-
docera : Daphniidae) from Taiwan by morphometric
and molecular analyses // Zool. stud. – 2012. – 51,
¹ 2. – P. 222–231.
35. Choudhury S., Dutta K. Study on the morphometric
variation in selected ichthyofauna under genus Pun-
tius Hamilton-Buchanan (Teleostei: Cyprinidae) //
J. Pharm. Biol. Sci. – 2013. – 5, ¹ 3. – P. 1–6.
36. Holthuis L.B. Shrimps and Prawns of the world. An
annotated catalogue of species of interest to fisheri-
es // FAO Species Catalogue. – Roma, 1980. – 125,
¹ 1. – 271 p.
37. Perez Farfante I., Kensley B. Penaeoid and Serges-
toid Shrimps and Prawns of the World, Keys and
Diagnoses for the Families and Genera // Memoires
du Museum national d’Histoire Naturelle. – Paris,
1997. – 175. – P. 1–233.
38. Chan T.Y. Shrimps and Prawns // The Living Marine
Resources of the Western Central Pacific / Eds K.E.
Carpenter, V.H. Niem. – Rome : FAO, 1998. –
P. 851–897.
39. Elliott N.G., Haskard K., Koslow J.A. Morphometric
analysis of orange roughy (Hoplostethus atlanticus)
off the continental slope of Southern Australia // J.
Fish. Biol. – 1995. – 46, ¹ 2. – P. 202–220.
40. Williams J.G.K., Kubelik A.R., Livak K.J. et al. DNA
polymorphisms amplified by arbitrary primers are use-
ful as genetic markers // Nucl. Acid. Res. – 1990. –
18, ¹ 22. – P. 6531–6535.
41. Jaccard P. Nouvelles recherches sur la distribution
florale // Bull. Soc. Vaud. Sci. Nat. – 1908. – 44. –
P. 223–270.
42. Rohlf F.J. Numeral Taxonomy and Multivariate
Analysis System, NTSYS-pc. Version 1.80. – New
York : Exeter Softeware, 1993.
43. Bookstein F.L. Morphometric Tools for Landmark
Data : Geometry and Biology. – Cambridge : Univ.
press, 1991. – 435 p.
44. Darwin C. On the origin of species by means of natu-
ral selection, or the preservation of favored, races in
the struggle for life. – London : John Murray, 1859. –
342 p.
45. Simpson G.G. Tempo and Mode in Evolution. –
New York : Columbia Univ. press, 1944. – 237 p.
46. Mayr E. Animal species and evolution. – Cambrigde :
Univ. press, 1963. – 797 p.
47. Hennig W. Phylogenetic Systematic. – Urbana:
Univ. press, 1966. – 263 p.
48. Eldredge N., Cracraft J. Phylogenetic patterns and
the evolutionary process. Method and theory in
comparative biology. – New York : Columbia Univ.
press, 1980. – 349 p.
49. Mayr E., Ashlock P.D. Principle of systematic zool-
ogy. – New York : McGraw-Hill, 1991. – 475 p.
50. Mulley J.C., Latter B.D.H. Genetic variation and
evolutionary relationships within a group of thirteen
species of penaeid prawns // Evolution. – 1980. –
34, ¹ 5. – P. 904–916.
51. Tam Y.K., Chu K.H. Electrophoretic study on the
phylogenetic relationships of some species of Penae-
us and Metapenaeus (Decapoda: Penaeidae) from the
South China Sea // Crustacean Biol. – 1993. – 13,
¹ 4. – P. 697–705.
52. Abolfazl Bayrami M., Shakunthala V. Molecular sys-
tematic of some shrimps based on allozyme data //
Res. Fish. Hydrobiol. – 2010. – 5, ¹ 2. – P. 152–158.
Received 08.05.13
<<
/ASCII85EncodePages false
/AllowTransparency false
/AutoPositionEPSFiles true
/AutoRotatePages /None
/Binding /Left
/CalGrayProfile (Dot Gain 20%)
/CalRGBProfile (sRGB IEC61966-2.1)
/CalCMYKProfile (U.S. Web Coated \050SWOP\051 v2)
/sRGBProfile (sRGB IEC61966-2.1)
/CannotEmbedFontPolicy /Error
/CompatibilityLevel 1.4
/CompressObjects /Tags
/CompressPages true
/ConvertImagesToIndexed true
/PassThroughJPEGImages true
/CreateJobTicket false
/DefaultRenderingIntent /Default
/DetectBlends true
/DetectCurves 0.0000
/ColorConversionStrategy /CMYK
/DoThumbnails false
/EmbedAllFonts true
/EmbedOpenType false
/ParseICCProfilesInComments true
/EmbedJobOptions true
/DSCReportingLevel 0
/EmitDSCWarnings false
/EndPage -1
/ImageMemory 1048576
/LockDistillerParams false
/MaxSubsetPct 100
/Optimize true
/OPM 1
/ParseDSCComments true
/ParseDSCCommentsForDocInfo true
/PreserveCopyPage true
/PreserveDICMYKValues true
/PreserveEPSInfo true
/PreserveFlatness true
/PreserveHalftoneInfo false
/PreserveOPIComments false
/PreserveOverprintSettings true
/StartPage 1
/SubsetFonts true
/TransferFunctionInfo /Apply
/UCRandBGInfo /Preserve
/UsePrologue false
/ColorSettingsFile ()
/AlwaysEmbed [ true
]
/NeverEmbed [ true
]
/AntiAliasColorImages false
/CropColorImages true
/ColorImageMinResolution 300
/ColorImageMinResolutionPolicy /OK
/DownsampleColorImages true
/ColorImageDownsampleType /Bicubic
/ColorImageResolution 1200
/ColorImageDepth -1
/ColorImageMinDownsampleDepth 1
/ColorImageDownsampleThreshold 1.50000
/EncodeColorImages false
/ColorImageFilter /DCTEncode
/AutoFilterColorImages true
/ColorImageAutoFilterStrategy /JPEG
/ColorACSImageDict <<
/QFactor 0.15
/HSamples [1 1 1 1] /VSamples [1 1 1 1]
>>
/ColorImageDict <<
/QFactor 0.15
/HSamples [1 1 1 1] /VSamples [1 1 1 1]
>>
/JPEG2000ColorACSImageDict <<
/TileWidth 256
/TileHeight 256
/Quality 30
>>
/JPEG2000ColorImageDict <<
/TileWidth 256
/TileHeight 256
/Quality 30
>>
/AntiAliasGrayImages false
/CropGrayImages true
/GrayImageMinResolution 300
/GrayImageMinResolutionPolicy /OK
/DownsampleGrayImages true
/GrayImageDownsampleType /Bicubic
/GrayImageResolution 1200
/GrayImageDepth -1
/GrayImageMinDownsampleDepth 2
/GrayImageDownsampleThreshold 1.50000
/EncodeGrayImages false
/GrayImageFilter /DCTEncode
/AutoFilterGrayImages true
/GrayImageAutoFilterStrategy /JPEG
/GrayACSImageDict <<
/QFactor 0.15
/HSamples [1 1 1 1] /VSamples [1 1 1 1]
>>
/GrayImageDict <<
/QFactor 0.15
/HSamples [1 1 1 1] /VSamples [1 1 1 1]
>>
/JPEG2000GrayACSImageDict <<
/TileWidth 256
/TileHeight 256
/Quality 30
>>
/JPEG2000GrayImageDict <<
/TileWidth 256
/TileHeight 256
/Quality 30
>>
/AntiAliasMonoImages false
/CropMonoImages true
/MonoImageMinResolution 1200
/MonoImageMinResolutionPolicy /OK
/DownsampleMonoImages true
/MonoImageDownsampleType /Bicubic
/MonoImageResolution 1200
/MonoImageDepth -1
/MonoImageDownsampleThreshold 1.50000
/EncodeMonoImages false
/MonoImageFilter /CCITTFaxEncode
/MonoImageDict <<
/K -1
>>
/AllowPSXObjects false
/CheckCompliance [
/None
]
/PDFX1aCheck false
/PDFX3Check false
/PDFXCompliantPDFOnly false
/PDFXNoTrimBoxError true
/PDFXTrimBoxToMediaBoxOffset [
0.00000
0.00000
0.00000
0.00000
]
/PDFXSetBleedBoxToMediaBox true
/PDFXBleedBoxToTrimBoxOffset [
0.00000
0.00000
0.00000
0.00000
]
/PDFXOutputIntentProfile (None)
/PDFXOutputConditionIdentifier ()
/PDFXOutputCondition ()
/PDFXRegistryName ()
/PDFXTrapped /False
/CreateJDFFile false
/Description <<
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
/BGR <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>
/CHS <FEFF4f7f75288fd94e9b8bbe5b9a521b5efa7684002000410064006f006200650020005000440046002065876863900275284e8e9ad88d2891cf76845370524d53705237300260a853ef4ee54f7f75280020004100630072006f0062006100740020548c002000410064006f00620065002000520065006100640065007200200035002e003000204ee553ca66f49ad87248672c676562535f00521b5efa768400200050004400460020658768633002>
/CHT <FEFF4f7f752890194e9b8a2d7f6e5efa7acb7684002000410064006f006200650020005000440046002065874ef69069752865bc9ad854c18cea76845370524d5370523786557406300260a853ef4ee54f7f75280020004100630072006f0062006100740020548c002000410064006f00620065002000520065006100640065007200200035002e003000204ee553ca66f49ad87248672c4f86958b555f5df25efa7acb76840020005000440046002065874ef63002>
/CZE <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>
/DAN <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>
/DEU <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>
/ESP <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>
/ETI <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>
/FRA <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>
/GRE <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>
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
/HRV (Za stvaranje Adobe PDF dokumenata najpogodnijih za visokokvalitetni ispis prije tiskanja koristite ove postavke. Stvoreni PDF dokumenti mogu se otvoriti Acrobat i Adobe Reader 5.0 i kasnijim verzijama.)
/HUN <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>
/ITA <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>
/JPN <FEFF9ad854c18cea306a30d730ea30d730ec30b951fa529b7528002000410064006f0062006500200050004400460020658766f8306e4f5c6210306b4f7f75283057307e305930023053306e8a2d5b9a30674f5c62103055308c305f0020005000440046002030d530a130a430eb306f3001004100630072006f0062006100740020304a30883073002000410064006f00620065002000520065006100640065007200200035002e003000204ee5964d3067958b304f30533068304c3067304d307e305930023053306e8a2d5b9a306b306f30d530a930f330c8306e57cb30818fbc307f304c5fc59808306730593002>
/KOR <FEFFc7740020c124c815c7440020c0acc6a9d558c5ec0020ace0d488c9c80020c2dcd5d80020c778c1c4c5d00020ac00c7a50020c801d569d55c002000410064006f0062006500200050004400460020bb38c11cb97c0020c791c131d569b2c8b2e4002e0020c774b807ac8c0020c791c131b41c00200050004400460020bb38c11cb2940020004100630072006f0062006100740020bc0f002000410064006f00620065002000520065006100640065007200200035002e00300020c774c0c1c5d0c11c0020c5f40020c2180020c788c2b5b2c8b2e4002e>
/LTH <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>
/LVI <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>
/NLD (Gebruik deze instellingen om Adobe PDF-documenten te maken die zijn geoptimaliseerd voor prepress-afdrukken van hoge kwaliteit. De gemaakte PDF-documenten kunnen worden geopend met Acrobat en Adobe Reader 5.0 en hoger.)
/NOR <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>
/POL <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>
/PTB <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>
/RUM <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>
/RUS <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>
/SKY <FEFF0054006900650074006f0020006e006100730074006100760065006e0069006100200070006f0075017e0069007400650020006e00610020007600790074007600e100720061006e0069006500200064006f006b0075006d0065006e0074006f0076002000410064006f006200650020005000440046002c0020006b0074006f007200e90020007300610020006e0061006a006c0065007001610069006500200068006f0064006900610020006e00610020006b00760061006c00690074006e00fa00200074006c0061010d00200061002000700072006500700072006500730073002e00200056007900740076006f00720065006e00e900200064006f006b0075006d0065006e007400790020005000440046002000620075006400650020006d006f017e006e00e90020006f00740076006f00720069016500200076002000700072006f006700720061006d006f006300680020004100630072006f00620061007400200061002000410064006f00620065002000520065006100640065007200200035002e0030002000610020006e006f0076016100ed00630068002e>
/SLV <FEFF005400650020006e006100730074006100760069007400760065002000750070006f0072006100620069007400650020007a00610020007500730074007600610072006a0061006e006a006500200064006f006b0075006d0065006e0074006f0076002000410064006f006200650020005000440046002c0020006b006900200073006f0020006e0061006a007000720069006d00650072006e0065006a016100690020007a00610020006b0061006b006f0076006f00730074006e006f0020007400690073006b0061006e006a00650020007300200070007200690070007200610076006f0020006e00610020007400690073006b002e00200020005500730074007600610072006a0065006e006500200064006f006b0075006d0065006e0074006500200050004400460020006a00650020006d006f0067006f010d00650020006f0064007000720065007400690020007a0020004100630072006f00620061007400200069006e002000410064006f00620065002000520065006100640065007200200035002e003000200069006e0020006e006f00760065006a01610069006d002e>
/SUO <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>
/SVE <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>
/TUR <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>
/UKR <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>
/ENU (Use these settings to create Adobe PDF documents best suited for high-quality prepress printing. Created PDF documents can be opened with Acrobat and Adobe Reader 5.0 and later.)
>>
/Namespace [
(Adobe)
(Common)
(1.0)
]
/OtherNamespaces [
<<
/AsReaderSpreads false
/CropImagesToFrames true
/ErrorControl /WarnAndContinue
/FlattenerIgnoreSpreadOverrides false
/IncludeGuidesGrids false
/IncludeNonPrinting false
/IncludeSlug false
/Namespace [
(Adobe)
(InDesign)
(4.0)
]
/OmitPlacedBitmaps false
/OmitPlacedEPS false
/OmitPlacedPDF false
/SimulateOverprint /Legacy
>>
<<
/AddBleedMarks false
/AddColorBars false
/AddCropMarks false
/AddPageInfo false
/AddRegMarks false
/ConvertColors /ConvertToCMYK
/DestinationProfileName ()
/DestinationProfileSelector /DocumentCMYK
/Downsample16BitImages true
/FlattenerPreset <<
/PresetSelector /MediumResolution
>>
/FormElements false
/GenerateStructure false
/IncludeBookmarks false
/IncludeHyperlinks false
/IncludeInteractive false
/IncludeLayers false
/IncludeProfiles false
/MultimediaHandling /UseObjectSettings
/Namespace [
(Adobe)
(CreativeSuite)
(2.0)
]
/PDFXOutputIntentProfileSelector /DocumentCMYK
/PreserveEditing true
/UntaggedCMYKHandling /LeaveUntagged
/UntaggedRGBHandling /UseDocumentProfile
/UseDocumentBleed false
>>
]
>> setdistillerparams
<<
/HWResolution [2400 2400]
/PageSize [612.000 792.000]
>> setpagedevice
|