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The Evolutionary History of the Rediscovered Austrian Population of the Giant Centipede Scolopendra cingulata Latreille 1829 (Chilopoda, Scolopendromorpha)


Abstract and Figures

The thermophilous giant centipede Scolopendra cingulata is a voracious terrestrial predator, which uses its modified first leg pair and potent venom to capture prey. The highly variable species is the most common of the genus in Europe, occurring from Portugal in the west to Iran in the east. The northernmost occurrences are in Hungary and Romania, where it abides in small isolated fringe populations. We report the rediscovery of an isolated Austrian population of Scolopendra cingulata with the first explicit specimen records for more than 80 years and provide insights into the evolutionary history of the northernmost populations utilizing fragments of two mitochondrial genes, COI and 16S, comprising 1,155 base pairs. We test the previously proposed hypothesis of a speciation by distance scenario, which argued for a simple range expansion of the species from the southeast, via Romania, Hungary and finally to Austria, based on a comprehensive taxon sampling from seven countries, including the first European mainland samples. We argue that more complex patterns must have shaped the current distribution of S. cingulata and that the Austrian population should be viewed as an important biogeographical relict in a possible microrefugium. The unique haplotype of the Austrian population could constitute an important part of the species genetic diversity and we hope that this discovery will initiate protective measures not only for S. cingulata, but also for its habitat, since microrefugia are likely to host further rare thermophilous species. Furthermore, we take advantage of the unprecedented sampling to provide the first basic insights into the suitability of the COI fragment as a species identifying barcode within the centipede genus Scolopendra.
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The Evolutionary History of the Rediscovered Austrian
Population of the Giant Centipede
Scolopendra cingulata
Latreille 1829 (Chilopoda, Scolopendromorpha)
Jan Philip Oeyen
*, Sebastian Funke
, Wolfgang Bo
, Thomas Wesener
1Department Arthropoda, Zoological Research Museum Alexander Koenig, Bonn, Germany, 2Department of Ophthalmology, University Medical Center of the Johannes
Gutenberg-University, Mainz, Germany
The thermophilous giant centipede Scolopendra cingulata is a voracious terrestrial predator, which uses its modified first leg
pair and potent venom to capture prey. The highly variable species is the most common of the genus in Europe, occurring
from Portugal in the west to Iran in the east. The northernmost occurrences are in Hungary and Romania, where it abides in
small isolated fringe populations. We report the rediscovery of an isolated Austrian population of Scolopendra cingulata with
the first explicit specimen records for more than 80 years and provide insights into the evolutionary history of the
northernmost populations utilizing fragments of two mitochondrial genes, COI and 16S, comprising 1,155 base pairs. We
test the previously proposed hypothesis of a speciation by distance scenario, which argued for a simple range expansion of
the species from the southeast, via Romania, Hungary and finally to Austria, based on a comprehensive taxon sampling from
seven countries, including the first European mainland samples. We argue that more complex patterns must have shaped
the current distribution of S. cingulata and that the Austrian population should be viewed as an important biogeographical
relict in a possible microrefugium. The unique haplotype of the Austrian population could constitute an important part of
the species genetic diversity and we hope that this discovery will initiate protective measures not only for S. cingulata, but
also for its habitat, since microrefugia are likely to host further rare thermophilous species. Furthermore, we take advantage
of the unprecedented sampling to provide the first basic insights into the suitability of the COI fragment as a species
identifying barcode within the centipede genus Scolopendra.
Citation: Oeyen JP, Funke S, Bo
¨hme W, Wesener T (2014) The Evolutionary History of the Rediscovered Austrian Population of the Giant Centipede Scolopendra
cingulata Latreille 1829 (Chilopoda, Scolopendromorpha). PLoS ONE 9(9): e108650. doi:10.1371/journal.pone.0108650
Editor: Valerio Ketmaier, Institute of Biochemistry and Biology, Germany
Received June 11, 2014; Accepted September 3, 2014; Published September 24, 2014
Copyright: ß2014 Oeyen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: The authors confirm that all data underlying the findings are fully available without restriction. All 43 sequences are available from the
GenBank database (accession numbers KJ812046-KJ812088). All sequence alignments and tables of uncorrected p-distances are within the paper and its
Supporting Information files.
Funding: Lab work and sequencing was funded by the Alexander-Koenig-Gesellschaft (AKG). Application for funding was submitted by TW. Grant-ID:
Scolopendra. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests: The authors have declared that no competing interests exist.
* Email:
The giant centipede Scolopendra cingulata Latreille, 1829 is
Europe’s largest centipede and the most common species of its
genus. It is famous for its voracious habits and painful bite as well
as its highly variable, often striking color pattern. Scolopendra is
the only European myriapod genus that can severely harm (e.g.
[1]), and in rare cases cause the death of humans [2,3].
Components of the potent venom were recently discovered to be
of potential medical significance as a pain reliever [4] and as an
inhibitor to the proliferation of different cancer types and bacteria
S. cingulata is widespread and common surrounding the
Mediterranean Sea [6] (Fig. 1), and, in the past, was divided into
three geographically distinct clades based on morphology: Western
Europe, Italy, and Eastern Europe [7,8]. While the species, in rare
cases, has been dispersed to Central Europe through commerce
(e.g., a specimen found in the city of Cologne, Germany; [9]), the
natural distribution of S. cingulata reaches its northern limit in
Romania, Hungary and Austria, where it occurs in small, isolated
populations (e. g. [10]). Multiple recent records exist from
Hungary [11,12] where it is listed as an endangered species and
receives special protection. In comparison, the Austrian Scolopen-
dra cingulata is all but forgotten, not even listed in recent species
distribution maps [6], despite the fact that its isolated populations
might be at least as endangered and localized as the populations in
Scolopendra cingulata in Austria
Scolopendra cingulata was mentioned as belonging to the
Austrian fauna by Latzel in 1880 [13]. His records, however, refer
to the Austro-Hungarian monarchy and these localities now lie in
Croatia and Hungary.
The first reference to the species occurring in the Lake Neusiedl
area in modern Austria was made by Attems in 1930 [10]. He
believed that the species ‘‘penetrates the Balkans up to Romania,
southern Hungary, and advanced through western Hungary to the
Leitha Mountains at the Lake of Neusiedl’’. This theory was later
refined by Franz [14,15], who characterized S. cingulata as a
typical relict form confined to steppe heathland, and to be found in
PLOS ONE | 1 September 2014 | Volume 9 | Issue 9 | e108650
Central Europe only in the Leitha Mts. close to the northern bank
of Lake Neusiedl. Szalay [16] argued against the proposed relict
scenario and claimed that further populations connecting the
scattered distribution would be found in future surveys. Next,
Wu¨rmli [17] cited these eastern, south-exposed slopes of the
Leitha Mts. as the northernmost reliable locality, but added a
second even more northern locality. However, the second locality,
viz. ‘‘Klosterneuburger Au’’ in Lower Austria, was added with a
question mark and without providing any actual specimen records.
The latest locality mentioned for the species within modern
Austrian borders is an isolated hill called Hackelsberg, which is
located between the Leitha Mts. and Lake Neusiedl. This locality
is mentioned as the only occurrence of S. cingulata in Austria by
Kasy in 1979 [18] and is again mentioned by Haider in 2008 [19],
both, again, without any specimen records. The old locality Zeiler
Berg was mentioned again as an extant locality by Ziegler et al.
[20] and referred to repeated findings of this centipede from 1981
onwards, made by an annual student field excursion to this region
by the ZFMK (see below) headed by one of us (WB). The most
recent reference is the exhaustive monograph on Austria’s
endemic plant and animal species compiled by Rabitsch & Essl
in 2009 [21]. Here, S. cingulata is not listed in the paragraph on
chilopods because it is not believed to be an Austrian endemic or
‘‘subendemic’’; next to 3 or 4 Cryptops species S. cingulata is only
briefly mentioned as one of a few widely distributed Scolopen-
dromorpha, ‘‘at only one single site in northern Burgenland’’ [22].
Generally, the Austrian population was forgotten or believed to be
extinct. In the time since the respective original publications, no
recent reports have confirmed the existence of the species at the
afore mentioned localities and it is, as already mentioned, exempt
from the most current revision of the distribution of old world
Scolopendra species [6].
Despite the fact that S. cingulata represents one of the iconic
European myriapod species with a wealth of studies of its ecology
(e.g. [23,24]), morphology (e.g. [8,25,26]), behavior (e.g.
[3,27,28]), and distribution (e.g. [6,29]), so far molecular studies
have only focused on the Greek island populations [30]. In these
studies the phylogeography of the species in the Aegean Sea could
be reconstructed using a molecular phylogeny of different island
populations of S. cingulata. Here, we widen the scope by clarifying
the evolutionary history of the rediscovered, strongly localized,
and potentially endangered population of S. cingulata in Austria
based on a molecular phylogeny, comparing samples from Austria,
Hungary, and Romania, including the first S. cingulata samples
from the European mainland examined to date. We aim to test the
speciation by distance hypothesis, stated by previous authors
[10,14,15], that S. cingulata reached its current relic area in
Austria via the Carpathians, through Romania and Hungary, by
testing for a correlation between genetic and geographic distance
between the different populations. Furthermore, we take advan-
tage of the broad intra-specific sampling to gain first basic insights
into the applicability of the COI and 16S fragments as species-
specific barcodes inside the genus Scolopendra.
Material and Methods
Taxon sampling
Austria. Specimens of the Austrian S. cingulata population
were annually watched, studied, and occasionally collected by one
of us (WB) at the single known site between 1981–2010, with a
handful of specimens stored over the years as vouchers in the
Figure 1. Distribution of
Scolopendra cingulata
.Modified after Lewis [6], showing countries where the species occurs, not exact area of
distribution. Numbers correspond to map numbers in Table 1 and question marks represent areas with ambiguous information.
Evolutionary History of the Austrian Scolopendra cingulata
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collections of the ZFMK. Permits for field studies and specimen
collection were granted by the local authorities (Amt der
Burgenla¨ndischen Landesregierung, Abt. 5 – Anlagenrecht,
Umweltschutz und Verkehr). In 2010 it was discovered (by WB
& TW) that the specimens from the Austrian population represent
the only records of Austrian Scolopendra in the last 80 years or
more, and its origin became the focus of research interests. To
infer the evolutionary history of the Austrian S. cingulata, genetic
material was collected from specimens from the population in
2011. To limit the impact on the presumably small population,
single legs were removed from seven adult specimens, which were
released alive. Legs of two additional adults, as well as an already
dead specimen, were collected from the same population in 2012.
Europe. Since no referenced sequences of S. cingulata from
the European mainland are available in GenBank (but.70 from
Greek islands [30]), additional specimens were analyzed. Because
of the supposed eastern origin of the Austrian populations, three
Hungarian specimens (Permit: Environmental Conservation Fund
No. 027798/2001) and old Museum samples (ZFMK) from
Romania, the Greek mainland, and Turkey were added (see
Table 1). To rule-out a Western origin of the Austrian population,
a sample from SW France was also included. Museum specimens
from Italy yielded no suitable DNA. Sequences of each of the main
Aegean groups (C1, C2, C3, see [30]) were added from GenBank
(Accession numbers: see Table 1).
Outgroups. Sequences from S. cretica Lucas, 1853 and S.
canidens Newport, 1844 were added from GenBank (Accession
numbers: see Table 1). Because no sequences are available on
GenBank, a Museum specimen (ZFMK) of S. oraniensis Lucas,
1846 from Morocco was also added to the analysis (Table 1).
The total dataset included 30 terminals for the COI (21 newly
added), 28 for the 16S (22 newly added), and 30 for the combined
dataset (22 newly added), respectively. Locality data (Table 1 and
fig. 1) is only given imprecisely because S. cingulata is actively
traded in the exotic pet market, and the continuous existence of
small fringe populations could be harmed by overzealous
DNA extraction, amplification, and sequencing
Total genomic DNA was extracted using the DNeasy Blood &
Tissue Kit (Qiagen; Valencia, CA, USA).
To study the evolutionary history of the Austrian S. cingulata
population, fragments of the mitochondrial cytochrome coxidase
subunit I (COI) encoding gene and the mitochondrial 16S rRNA
(16S) encoding gene were amplified. Both gene fragments have
previously been successfully applied to study centipede evolution at
both the genus (e.g. [31–33]) and species-level (e.g. [30]). Since the
16S fragment only provided low resolution, we decided not to
include slower evolving nuclear rDNA genes in this study.
The 16S fragment was amplified using the 16Sa/16Sb primer
pair [34]. The COI fragment was amplified from samples Sco01-
10 (Austria) using the primer pair LCO1490/HCO2198 [35]. For
the remaining samples Nancy [36] was used as an alternative
reverse primer. Attempts were also made to amplify the region
with the HCOoutout primer [37,38], but no results of sufficient
quality could be obtained even though a wide range of PCR-
programs were applied. All polymerase chain reactions (PCR)
were carried out using the QIAGEN Multiplex PCR Kit and a
T3000 Thermocycler (Biometra). All PCR setups included a
positive and negative control. Detailed descriptions of temperature
profiles and PCR-mixtures can be found in a previous study [39].
The PCR products were inspected on a 1.5% agarose gel and
purified using the QIAquick PCR Purification Kit (Qiagen,
following the kit protocol, Valencia, CA, USA). Both strands were
sequenced by Macrogen (Macrogen Europe Laboratory, Amster-
dam, The Netherlands), using the PCR primers. Sequencing reads
were assembled and edited using Geneious 6.0.6 (Biomatters) and
Seqman II (DNASTAR, Inc.). Sequence identities were confirmed
with BLAST searches [40]. All new sequences were deposited in
GenBank (see Table 1 for accession numbers).
All sequences were aligned using the MUSCLE algorithm [41]
under the default settings as implemented in Geneious (Biomatters)
and edited by hand. Missing ends were filled with N’s. The
following sites were deleted from the 16S dataset prior to analysis
to remove regions of ambiguous homology, mostly regarding the
outgroups: 487, 468, 351–353, 345–346, 332–336, 348, 149–150,
20, 1–7. The final alignments consisted of 508 bp (16S), 647 bp
(COI) and 1155 bp in the combined dataset. Fasta files of all
alignments and tables containing the uncorrected p-distances for
both genes can be found in the supplementary material (Tables S1,
S2, and Alignments S1, S2, S3).
Sequence analysis
In all maximum likelihood analyses the dataset was analyzed
using the model suggested by the Bayesian Information Criterion
(BIC), which was computed by the model test implemented in
MEGA 5.1 [42]. The models with the highest fit were HKY+G
[43] for the 16S dataset (BIC = 3107.8) and GTR+G [44] for the
COI (BIC = 4855.9) and the combined dataset (BIC = 11724.8).
In order to assess the phylogenetic information in our 16S and
COI datasets, a likelihood mapping [45] was conducted with
TREE-PUZZLE 5.2 [46].
Maximum Likelihood phylogenetic analysis
All maximum likelihood (ML) analyses were conducted in Mega
5.1 [42]. The initial trees were made by Neighbor joining [47], the
heuristic search was conducted with the Nearest Neighbor
Interchange algorithm [48] and nodal support values were
assessed with 1000 bootstrap pseudoreplicates. The tree obtained
by the maximum likelihood analysis of the combined alignments
was used for all further discussion and interpretation of the results
(Fig. 2C).
Maximum Parsimony phylogenetic analysis
All Maximum Parsimony (MP) analyses were performed in
PAUP* 4.0b10 [49] using the TBR algorithm. Starting trees were
obtained via stepwise addition and nodal support was estimated
with 1000 bootstrap replicates (unlimited number of trees kept at
each replicate). The combined dataset included a total of 304 (16S:
124, COI: 180) parsimony informative characters. For the 16S
dataset, 1676 shortest trees with 289 steps were found. For the
COI dataset, 175714 shortest trees with 568 steps were found. The
analysis of the combined dataset resulted in 2101 shortest trees
with 867 steps. Strict consensus trees were produced for all
datasets (trees not shown). Nodal support values of the MP
bootstrap analysis are displayed in Figure 2C.
Bayesian phylogenetic analysis
Bayesian inference (BI) was conducted using MrBayes 3.1.2
[50]. Each dataset was analyzed with the model suggested by the
model test implemented in MEGA 5.1 [42], as described above.
The combined dataset was partitioned to allow unlinked models
for the two genes. The model parameters (priors) were left unfixed
to allow estimation from the dataset, as suggested by the MrBayes
manual. The analysis was performed using 3,000,000 Monte
Evolutionary History of the Austrian Scolopendra cingulata
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Table 1. Overview of samples included in the present study, with numbers corresponding to the map (Fig. 1), voucher numbers, locality information and accession numbers.
Accession numbers
#Map Sample ID Voucher ZFMK #Species Locality COI 16S
1 1 Austria-1 (1) ZFMK-Sco-1 S. cingulata Austria, Brugenland, Leitha Mts. KJ812067 KJ812046
2 1 Austria-2 (2) ZFMK-Sco-2 S. cingulata Austria, Brugenland, Leitha Mts. KJ812068 KJ812047
3 1 Austria-3 (3) ZFMK-Sco-3 S. cingulata Austria, Brugenland, Leitha Mts. KJ812069 n/a
4 1 Austria-4 (4) ZFMK-Sco-4 S. cingulata Austria, Brugenland, Leitha Mts. KJ812070 KJ812048
5 1 Austria-5 (5) ZFMK-Sco-5 S. cingulata Austria, Brugenland, Leitha Mts. KJ812071 KJ812049
6 1 Austria-6 (6) ZFMK-Sco-6 S. cingulata Austria, Brugenland, Leitha Mts. KJ812072 KJ812050
7 1 Austria-7 (7) ZFMK-Sco-7 S. cingulata Austria, Brugenland, Leitha Mts. KJ812073 KJ812051
8 1 Austria-8 (8) ZFMK-Sco-8 S. cingulata Austria, Brugenland, Leitha Mts. KJ812074 KJ812052
9 1 Austria-9 (9) ZFMK-Sco-9 S. cingulata Austria, Brugenland, Leitha Mts. KJ812075 KJ812053
10 1 Austria-10 (10) ZFMK-Sco-10 S. cingulata Austria, Brugenland, Leitha Mts. KJ812076 KJ812054
11 1 Austria-11 (11) Myr 01591 S. cingulata Austria, Brugenland, Leitha Mts. n/a KJ812055
12 1 Austria-12 (12) Myr 01592 S. cingulata Austria, Brugenland, Leitha Mts. n/a KJ812056
13 2 Hungary-1 (13) Myr 01559 S. cingulata Hungary, Ve
´rtes Mts., Csa
KJ812077 KJ812057
14 2 Hungary-2 (14) Myr 01560 S. cingulata Hungary, Ve
´rtes Mts., Csa
KJ812078 KJ812058
15 2 Hungary-3 (15) Myr 01561 S. cingulata Hungary, Ve
´rtes Mts., Csa
KJ812079 KJ812059
16 3 Greece_Kavala-1 (16) ZFMK-Sco-14 S. cingulata Greece, Kavala KJ812080 KJ812060
17 3 Greece_Kavala-2 (17) Myr 00585 S. cingulata Greece, Kavala KJ812081 n/a
18 4 Greece_Port-Lagos-1 (18) ZFMK-Sco-13 S. cingulata Greece, Nestos Delta, Port Lagos KJ812082 KJ812061
19 5 Greece_Port-Lagos-2 (19) ZFMK-Sco-15 S. cingulata Greece, Nestos Delta, Port Lagos KJ812083 KJ812062
20 6 *Greece_Nisyros (20) n/a S. cingulata Greece, Nisyros JN688371 JN688421
21 7 *Greece_Koufonisi (21) n/a S. cingulata Greece, Koufonisi JN688365 JN688413
22 8 *Greece_Paros (22) n/a S. cingulata Greece, Paros JN688377 JN688427
23 9 *Greece_Anafi (23) n/a S. cingulata Greece, Anafi JN688350 JN688398
24 10 *Greece_Amorgos (24) n/a S. cingulata Greece, Amorgos JN688349 JN688397
25 11 Romania (25) ZFMK-Sco-11 S. cingulata Romania, Anina KJ812086 KJ812065
26 12 Turkey_Troy (26) ZFMK-Sco-12 S. cingulata Turkey, Troy KJ812084 KJ812063
27 13 Turkey_Izmir (27) Myr 00583 S. cingulata Turkey, Izmir KJ812085 n/a
28 14 France (28) Myr 01593 S. cingulata France, Banyuls-sur-mer KJ812087 KJ812064
29 15 oraniensis (29) Myr 00568 S. oraniensis Marokko, Prov. Nador, Atlas Mts. KJ812088 KJ812066
30 16 cretica (30) n/a S. cretica Greece, Crete JN688393 JN688440
31 17 *canidens (31) n/a S. canidens Greece, Serifos JN688394 JN688441
32 18 *canidens (32) n/a S. canidens Greece, Sifnos JN688442 n/a
Sequences downloaded from GenBank are marked with an asterisk.
Evolutionary History of the Austrian Scolopendra cingulata
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Carlo Markov Chain (MCMC) generations of three hot and one
cold chain in two parallel runs, sampling trees every 100th
generation. The likelihood values for the parallel runs were
inspected manually and the generations prior to a stable value
were discarded as burn-in. The burn-in was set to 3600, 3100 and
8400 generations for the COI, 16S and concatenated dataset,
respectively. Nodal support values are displayed in Figure 2C.
Analysis of the Evolutionary History of the Austrian
S. cingulata population
In order to further test the hypothesis of a speciation by distance
scenario, as stated by previous authors [10,14,15], we compared
the genetic and geographic distances between the Austrian S.
cingulata and those from France, Greece, Turkey, Romania and
The S. cingulata COI sequences and geographic distances were
pooled according to populations (Tab. 2) and Kendall’s Tau
correlation test [51] was performed, as the data was unsuitable for
a Mantel test [52]. Kendall’s Tau allows to test for a correlation
between two variables where the measurements are not equidis-
tant and the data is non-parametric. To assess whether the data
are uncorrelated or not, the two-tailed probability test was also
performed. All tests were performed in PAST [53].
Figure 2. Hypothetical relationships of the northern
Scolopendra cinuglata
and phylogenetic tree recovered in maximum likelihood
analysis. A: The hypothetical relationships of the northern populations as previously stated by Attems [10] and Franz [14,15]. B: Adult Scolopendra
cingulata specimen from the Austrian population in situ. Photo by Dr. Wolfram Freund. C: Maximum likelihood tree of the combined COI and 16S
dataset. Numbers represent nodal support values from the maximum likelihood (1000 bootstrap replicates), maximum parsimony analysis (1000
bootstrap replicates) and posterior probabilities from the Bayesian inference (ML/MP/BI). Sequences from GenBank marked with single asterisk in
front of name. Samples with two asterisks after name include only the COI sequence. Numbers in parenthesis correspond to sample numbers in
Table 1.
Evolutionary History of the Austrian Scolopendra cingulata
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Barcode evaluation
To provide preliminary insights into the suitability of the COI
fragment as a species-delimiting barcode, the frequency distribu-
tion of all pairwise uncorrected p-distances were analyzed. If the
COI fragment is suitable for species identification within
Scolopendra, a (barcode) gap should exist between the inter- and
intra-specific distances [54,55].
Sequence Data
The sequencing was successful for most specimens, with the
exception of two (one from Izmir, Turkey; and one from Kavala,
Greece) of which only the COI was obtained, and of two
specimens from Austria of which only the 16S sequence could be
In the COI dataset, the A, T, C and G frequencies were 0.35,
0.27, 0.22 and 0.17, and in the 16S dataset, they were 0.30, 0.39,
0.09 and 0.22, respectively. The sequence composition in our COI
dataset shows a clear bias towards A and T, which has been shown
to be common within chilopods [56] and arthropods in general
The likelihood-mapping showed a higher amount of phyloge-
netic information content in the COI dataset than in the 16S
(Figs. 3A, B). The 16S analysis resulted in 24.0% unresolved trees
and a total of 11.2% partially resolved trees (Fig. 3B). The COI
analysis, on the other hand, resulted in only 12.7% unresolved
trees and a total of 6.1% partially resolved trees (Fig. 3A).
Molecular Phylogenetic Analyses
The trees obtained by the maximum likelihood analysis of the
combined dataset, with the added support values of the MP and
Bayesian analyses, are utilized for the presentation of results
(Fig. 2C).
In the analysis of the combined dataset, the S. cingulata samples
form a well-supported (ML = 96%, MP = 100%, PP = 100%) clade
against the out-group (S. cretica,S. canidens,S. oraniensis)
(Fig. 2C). The genetic distances between S. cingulata and its three
congeners are high (COI: 13.5–16.8%, 16S: 19.3–23.0%). In the
out-group, S. oraniensis branches off basally, where S. cretica and
S. canidens form a group (88% ML support). S. oraniensis seems
to be only slightly more closely related to S. canidens (uncorr. p-
dist.: COI: 14.5%, 16S: 22.7%), than to S. cretica (uncorr. p-dist.:
COI: 15.1%, 16S: 20.9%).
Within S. cingulata the basal-most branch consists of a well-
supported group (99/100/100) containing two Greek island
specimens, representing group C3 of previous analyses [30]. The
next split in the tree places the western European specimen outside
the only weakly supported (57% ML), clade of the remaining
samples (Fig. 2C). Within the clade, the sample from Nisyros (C1,
[30]) stands basally in a well-supported (56/66/100) group with
the two specimens from Turkey. The sister-group to the Greek-
Turkish clade is poorly supported. Inside the latter, the Austrian S.
cingulata represent the basal-most group. The group’s monophyly
receives strong support (98/100/93) and it contains only a single
haplotype in both the COI and 16S gene. The sister-group of the
Austrian S. cingulata is a clade consisting of specimens from
Hungary, Romania and Greece (Fig. 2C). Basally, the Hungarian
specimens form a well-supported monophyletic clade (100/99/99),
while their sister-groups are less well supported. The three
Hungarian samples, from localities less than 1 km apart, display
different COI haplotypes with small genetic distances (COI: 0.6–
1.0%, 16S: 0.0%). The first weakly supported (57/-/-) split within
the sister-group to the Hungarian samples, places the Romanian
sample outside of a clade containing the remaining Greek samples.
Within the Greek samples (excluding the basal Greek Island C3
and the Turkish-Nisyros C1), three well-supported clades can be
distinguished: (1) one specimen (red legged, Fig. 4A) from Port
Lagos, which forms the sister-group to (2) the island samples C2
(97/100/100), and (3) the well-supported (87/92/100) Greek
mainland specimens from Kavala and Port Lagos (yellow legged,
Fig. 4B).
Evolutionary History of the Austrian S. cingulata
The speciation by distance scenario, as suggested by Attems
[10] and Franz [14,15], postulates that S. cingulata could have
reached Austria from an eastern refugium or point of origin via
Hungary and Romania. A positive correlation between the
geographic and genetic distance, as would be expected under said
scenario, could not be proven. Although a weak positive
Table 2. Geographic and genetic distances (COI, uncorrected p) between the Austrian (Map #1) and all other populations.
Map #Localities Distance [km] Distance COI [%]
2 Hungary, Ve
´rtes Mts. 138 4,4
3 Greece, Kavala 989 2,4
4 Greece, Port Lagos I 1016 2,3
5 Greece, Port Lagos II 1016 2,1
6 Greece, Nisyros 1526 6,0
7 Greece, Koufonisi 1422 2,5
8 Greece, Paros 1395 2,5
9 Greece, Anafi 1488 5,8
10 Greece, Amorgos 1444 6,0
11 Romania 504 2,9
12 Turkey, Troy 1151 3,5
13 Turkey, Izmir 1353 3,1
14 France 1228 5,8
Evolutionary History of the Austrian Scolopendra cingulata
PLOS ONE | 6 September 2014 | Volume 9 | Issue 9 | e108650
correlation was detected (tau = 0.34213), the probability test failed
to support this scenario (p = 0.1035).
Barcode evaluation
A clear gap was found between the intra- and interspecific
distances (Fig. 3C). The average intraspecific distance was 6.4%
and the highest was 9.1% between the single French specimen and
one specimen from the Greek island Nisyros. The average
interspecific distance was 14.8% and the lowest was 13.5%
between one of the Hungarian S. cingulata specimens and S.
cretica, as well as between S. cretica and the S. canidens specimen
from Sifnos.
Sequence Data – Barcode evaluation
Our analysis of the COI dataset regarding the suitability of the
sequence as a species-delimiting barcode showed a clear ‘‘barcode
gap’’ between the intra- and interspecific distances. Such a gap
was also reported for the North-African representatives of the
lithobiid genus Eupolybothrus, with the lowest interspecific
distance of 16.61% and intraspecific distances of 1.4% and 0.3%
[33]. However, a finer geographical sampling of all taxa would be
necessary to validate our findings as high intraspecific distances
have been reported for the New Caledonian endemic species
Cryptops pictus, with a divergence of up to 23.8% [32], as well as
Figure 3. Results from likelihood mapping and barcode-gap analyses. A: Likelihood Mapping for COI dataset. B: Likelihood mapping for the
16S dataset. C: Barcode-gap analysis: Frequency distribution of the pairwise uncorrected p-distances of the COI sequences. Orange bars show
intraspecific distances and yellow bars represent interspecific distances.
Figure 4. Sympatrical
Scolopendra cingulata
color morphs from
Port Lagos (Greece),
.A:Red legged morph with black body.
B: Yellow legged morph with green-brown body.
Evolutionary History of the Austrian Scolopendra cingulata
PLOS ONE | 7 September 2014 | Volume 9 | Issue 9 | e108650
several instances of overlapping inter- and intraspecific distances in
Bavarian Chilopoda [56]. The latter was suggested to be due to
possible cryptic species or alternatively long separated haplotypes,
which would concur with the findings of Wiemers & Fidler [61].
They showed that barcode gaps could be artefacts resulting from
incomplete geographical sampling of widespread species. Using
the 16S fragment might pose a better option, as the highest
intraspecific divergence within our S. cingulata samples was only
4.5%, much lower than in the COI (9.1%), and the lowest
interspecific divergence was still at 10.2%, similar to the one of the
COI dataset (13.5%), between the closely related S. cretica and S.
canidens, and much higher between the other species (19.3–
23.0%). The 16S fragment would likely increase the accuracy of
the barcode, but at the cost of the population genetic insight.
Evolutionary History of the Austrian S. cingulata
The expected hypothetical phylogenetic tree (Fig. 2A), accord-
ing to the evolutionary hypothesis by past authors [10,14,15],
could not be recovered in any of the conducted analyses of the
mitochondrial genes (Fig. 2C). Although the most interesting splits
received weak statistical support in our phylogenetic analyses, it
remains clear that the Austrian, Hungarian, Romanian, and
northern Greek specimens are closely related. The study by
Simaiakis et al. [30] recovered the split between the eastern
Aegean Islands (C1) and the northern and central Cyclades (C2) to
most likely have happened approximately 10.12 Mya. In our
analysis of the combined COI and 16S datasets, the Austrian
population is the first to split from the branch leading to the
representatives of the C1 group (Fig. 2C). This might indicate that
the lineage of the Austrian population was established much
earlier than the possible repopulation of the Carpathian Basin
following the last glacial period. Furthermore, the fact that both of
the postulated closest relatives to the Austrian Scolopendra
population, the Hungarian and Romanian specimens, have a
lower genetic distance to the northern Greek samples (Port Lagos)
than to each other or to the Austrian population indicates that the
underlying pattern is more intricate than what can be explained by
a single range expansion of S. cingulata population founders out of
a Mediterranean refugium.
Our study failed to significantly prove the postulated speciation
by distance scenario suggested by Attems [10]. This implies that
more complex mechanisms and/or events preceding the last
glacial period could have shaped the northernmost distribution of
Scolopendra cingulata in Europe. Multiple independent recoloni-
zations would correspond with the view of Varga [62], who
concludes that populations from multiple small meso- and
microclimatically favourable sites at the fluctuating borderlines
of the Mediterranean refugial and periglacial belts played a
significant part in the postglacial repopulation of the Carpathian
Basin in several insect groups. Though this scenario of multiple
recolonizations seems probable, further and denser sampling,
especially of geographically close western populations and the
populations in the regions surrounding the former periglacial belts,
would be necessary to confirm our theory, as the current sampling
does not provide the resolution required to draw any firm
A valuable relict in a microrefugium: The last habitat of
the Austrian S. cingulata
Franz [14,15] proposed that the Austrian and Hungarian S.
cingulata populations are relicts of a previous wider distribution
during the post-glacial climatic optimum, which later became
isolated because of the following cooler climate and the expansion
of the forest. Szalay [16] on the other hand expected further
populations to be found, even connecting the populations to the
main area of distribution. This does not seem to be the case, as
such connections still have not been found after numerous
excursions to the area over the span of 30 years. Furthermore,
many of the cited localities in the literature are, at best,
implausible. The locality Klosterneuburger Au in Lower Austria,
mentioned by Wu¨rmli [17] seems absolutely unlikely to be a
suitable habitat for this thermophilous centipede, as ‘‘Au’’ means
the gallery forest along the Danube River close to the city of
Klosterneuburg. The locality at the Hackelsberg, as mentioned by
Haider [19], is also dubious. The accompanying photograph
shows a specimen from a doubtlessly Mediterranean rather than
Austrian population; it is actually from southern France as
communicated to us by the photographer, F. Geller-Grimm. All
of the above mentioned accounts are lacking specimen records,
and can therefore not be validated. Therefore, the Austrian, and to
some extent the Hungarian, Scolopendra cingulata populations
should be viewed as biogeographical relicts (sensu Lomolino et al.
The Austrian population, despite its low genetic variation,
represents a completely unique haplotype within the species
(Splitstree analysis: Data not presented). It has been shown that
peripheral relict populations of widespread species can harbor
unique genetic information [64] and that adaptations, which were
gained during the range expansions, are lost when the range
becomes restricted [65]. Additionally, relict populations might also
be important during future range expansions or shifts, enabling S.
cingulata to colonize a large area faster than what would be
possible through diffusional migration along a single expanding
front [66]. Thus, even though S. cingulata shows a widespread
distribution on a continental scale, the Austrian population, which
is a significant part of the genetic diversity, could be important to
the future survival of the species and should therefore be protected.
The small area inhabited by the Austrian population, an
exposed southern slope with scattered boulders of varying sizes,
should probably be considered a microrefugium, which can be
defined as a small area with local favorable environmental features
in which small populations can survive outside their main
distribution area, protected from the unfavorable regional
environmental conditions [67,68]. Favorable microclimatical
conditions and the neglect by farmers have probably allowed the
isolated population of the species to survive. This is supported by
the syntopical occurrence of protected thermophilous vertebrates
(e.g. Lacerta viridis, Zamenis longissimus) and several thermoph-
ilous insects, including the rare ground beetle Carabus hungaricus,
Mantis religiosa,Platycleis grisea, and, historically, Sago pedo.Itis
also likely that the unique habitat presently hosts further
thermophilous taxa. Even if the climatic conditions should change,
the site is still likely to retain a special microclimate relative to the
surroundings and will therefore possibly remain as a microrefu-
gium for a new set of species [66]. Consequently, the habitat is not
only worthy of protection to secure the current S. cingulata
population, but also to protect further species now and in the
Study of the European mainland S. cingulata populations
versus other S. cingulata studies
Minelli [7] proposed that S. cingulata populated southern
Europe (Iberian-, Italian and the Balkan Peninsula) quite recently.
In contrast, a later study based on distributional patterns [29]
suggested that the species differentiation in the Mediterranean
Basin happened less than 5.5 Mya, or, alternatively, between 9
and 12 Mya via either the Balkans or northern Africa. A recent
study based on molecular data supports the latter view, suggesting
Evolutionary History of the Austrian Scolopendra cingulata
PLOS ONE | 8 September 2014 | Volume 9 | Issue 9 | e108650
the time of divergence for two main lineages of Aegean S.
cingulata to have been approximately 10.5 Mya years ago [30].
However, our data suggests that the Aegean islands were most
likely colonized from multiple directions as supported by the fact
that the representatives from the C2 group cluster within the
Greek mainland samples and that the C1 sample clusters with the
Turkish mainland samples (Fig. 2C). This could imply that the
formation of the mid-Aegean trench might be irrelevant to the
C1/C2 split, rendering the calibration of the phylogeny in the
previous analysis [30] inaccurate.
An additional study of S. cingulata populations based on
morphometric data revealed an east-west gradient within the
species [8], suggesting a colonization of central and southern
Europe from the easternmost parts of its range (Asia Minor and
Middle East) via the Balkans and Northern Africa. Our analyses
support the notion of separate colonization events, because the
French sample is recovered in a basal position relative to the
eastern European samples. If the French population had
originated via the Balkans, a closer relationship with the Austrian
or Greek populations would have been expected. However, given
the limitations of the current sampling, our results are concurring
with but not corroborating the recently proposed multiple
colonizations of the European continent [8]. Regrettably, the
samples used for the present study were for the most part juveniles,
prohibiting a morphometric evaluation.
Two sympatric, unrelated different color morphs in Greek
S. cingulata
The two Greek mainland samples from Port Lagos were
animals of two different color morphs; one had red legs and a
black body (Fig. 4A) while the other had yellow legs with a green-
brown body (Fig. 4B). Such extreme color variation is not rare in
Scolopendra species. For example, Shelley [69] reports that S.
viridis in North America ranges from a solid green to yellow in
variations with longitudinal or transverse stripes. However, the
genetic basis of these variations is completely unknown. It is
especially interesting that the two sympatrically occurring morphs
do not seem to be each other’s closest relatives. The gray animal
clusters with the other Greek mainland samples from Kavala,
which were brown with yellow legs, and the red-black animal is
resolved as the sister taxon to the group containing the Greek
mainland samples and the samples from the Greek islands Paros
and Koufonisi (Fig. 2C). Further sampling would be required to
confirm if there is any phylogenetic information behind the
different morphs or if the variation is also present within one
lineage. Presently, we can only conclude that the differentiation is
clearly within the intra-specific range, since the divergence
between the two samples is only 2.3% (COI).
Analysis problems
The likelihood-mapping analyses we conducted show that the
16S fragment did not provide much phylogenetic information on
the intraspecific level, as a large portion of the trees remained
completely or at least partially unresolved. This is also evident in
the trees produced by our phylogenetic analyses, where the dataset
only provides some resolution in the most basal splits. The lack of
intraspecific variation (highest divergence of 4.3%) prompted us to
omit the gene from the analysis of the evolutionary history (but not
the phylogeny, see Fig. 2C). A faster evolving gene than COI or
16S is needed to further elucidate the evolutionary history of the
northernmost S. cingulata and for future studies in the genus
Scolopendra at the species level. While fragments of the 12S
[30,70] and 28S gene [70–73] have been employed in previous
centipede studies, these genes seem to be even slower evolving
than 16S. ITS (internal transcribed spacer) might be a future
alternative, but it does not seem to provide more species level
information than the COI-fragment [32].
Unfortunately, a microsatellite study – often the method of
choice [74] for population genetic studies in insects (e.g. [75]) and
vertebrates (e.g. [76]) – has never been conducted in Chilopoda. A
cheaper and easier alternative might be using AFLPs since the
method does not require specific primers or any previous
knowledge about the sequences [74,77]. However, an AFLP study
is not possible with old museum specimens. As previously
mentioned, a more fine-tuned and denser sampling across the
whole distributional range would also vastly improve the
conclusiveness of our analyses.
To test the hypothesis of multiple independent colonization
events and elucidate the phylogeography of the northernmost
populations of Scolopendra cingulata further, a finer geographic
taxon sampling as well as the application of other molecular
markers, as discussed above, is absolutely essential. Including
further peripheral populations will be difficult, as they are
extremely scattered and often restricted to very small areas.
Within Asturia S. cingulata is only known from the sample locality
(which is ,1000 m
) and the distribution in Hungary was only
revised recently [11,12,78], so that precise localities are available.
Such extremely localized fringe populations are difficult to localize
and sample. However, including further samples from the main
distribution area would be very interesting, since several
thermophilous taxa in the Carpathian basin show connections to
the Balkans, southern Russia and Asia Minor [79].
Supporting Information
Table S1 Uncorrected p-distances of 16S alignment.
Computed with Mega5 [42].
Table S2 Uncorrected p-distances of COI alignment.
Computed with Mega5 [42].
Alignment S1 Muscle [41] alignment of S. cingulata 16S
Alignment S2 Muscle [41] alignment of S. cingulata COI
Alignment S3 Muscle [41] alignment of concatenated S.
cingulata 16S and COI sequences.
The authors would like to thank Claudia Etzbauer for her help in the
molecular laboratory of the ZFMK. We also thank the students of the
Neusiedlersee excursions for their collection efforts, Rainer Hutterer for the
collection and contribution of the Moroccan specimen and La´zlo´Da´nyi
(Hungarian Natural History Museum) for providing us with samples from
the Hungarian populations. We also thank the local authorities in Austria
(Amt der Burgenla¨ ndischen Landesregierung) for granting collection
permits, as well as F.Geller-Grimm (Natural History Museum Wiesbaden),
who confirmed to us that his photograph of a S. cingulata published in the
note by Haider [19] was not at all taken in Austria but in southern France.
We thank Zoltan Korsos (Hungarian Natural History Museum) for his help
with the literature and comments on the topic. Lastly, we would also like to
thank Paul B. Frandsen (Rutgers University, USA) and Jeanne Wilbrandt
Evolutionary History of the Austrian Scolopendra cingulata
PLOS ONE | 9 September 2014 | Volume 9 | Issue 9 | e108650
(ZFMK) for proofreading and giving constructive comments on the
manuscript, as well as the two anonymous reviewers for their help in
improving the manuscript.
Author Contributions
Conceived and designed the experiments: JPO WB TW. Performed the
experiments: JPO. Analyzed the data: JPO TW. Contributed reagents/
materials/analysis tools: JPO SF WB TW. Wrote the paper: JPO SF WB
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Evolutionary History of the Austrian Scolopendra cingulata
PLOS ONE | 11 September 2014 | Volume 9 | Issue 9 | e108650
... The genetic distance among Scolopendra species ranges from 15.9-24.4% in .0% for European Scolopendra by Oeyen et al. (2014)). Comparing with different genera from the same/another subfamily, the distances are between 21.6-28. ...
... The two species are morphologically similar despite their markedly disjunct distributions, i.e., S. cingulata in the Mediterranean versus S. japonica in East Asia (Table 8). However, exploration of microrefugia of populations of S. cingulata during glacial maxima in Europe (Simaiakis et al. 2012, Oeyen et al. 2014) and a record of S. japonica in the northern part of Laos could indi cate that these two species may be more widespread than previously recognised. How ever, distributional data for S. japonica are patchy due to incomplete faunistic surveys in several parts in Asia. ...
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The centipede genus Scolopendra in mainland Southeast Asia is reviewed taxonomically based on morphological characters, informed by a molecular phylogenetic analysis using sequences from three mitochondrial and nuclear genes (COI, 16S rRNA and 28S rRNA). Eight nominal species of Scolopendra, namely S. morsitans Linnaeus, 1758, S. subspinipes Leach, 1816, S. dehaani Brandt, 1840, S. multidens Newport, 1844, S. calcarata Porat, 1876, S. japonica Koch, 1878, S. pinguis Pocock, 1891, and S. dawydoffi Kronmüller, 2012, are redescribed together with some revision of type materials. Geographical variation in each species has been compiled with reference to samples that span their distribution ranges in Southeast Asia and some parts of neighbouring areas such as East Asia, the Indian Ocean, and Africa. Comparative study of traditional taxonomic characters from external morphology provides further information to distinguish some closely related species. Scolopendra cataracta Siriwut, Edgecombe & Panha, sp. n., is described from the southern part of Laos, with additional records in Thailand and Vietnam. The phylogenetic framework for Southeast Asian Scolopendra recognizes S. calcarata + S. pinguis, S. morsitans, and a S. subspinipes group that unites the other six species as the main clades. Within the S. subspinipes group, two monophyletic groups can be distinguished by having either slender or short, thick ultimate leg prefemora and different numbers of apical spines on the coxopleuron. Scolopendra arborea Lewis, 1982, is placed in subjective synonymy with S. dehaani. A survey of external morphology of the genital segments confirms its potential for improving species identification in Scolopendra. Some observations on biology and behaviour are recorded based on field surveys in this area.
... There are only a handful of barcoding and phylogenetic studies applying molecular data of Scolopendromorpha worldwide (Murienne et al. 2010;Simaiakis et al. 2012;Vahtera et al. 2012Vahtera et al. , 2013Joshi and Edgecombe 2013;Oeyen et al. 2014;Siriwut et al. 2015). For Cryptops, there is only a singular molecular study utilizing barcoding genes and it deals with tropical pacific island species (Murienne et al. 2011). ...
... Barcoding studies inside the Scolopendromorpha consecutively revealed large interspecific distances (Simaiakis et al. 2012;Joshi and Edgecombe 2013;Oeyen et al. 2014;Siriwut et al. 2015). The only study involving Cryptops (Murienne et al. 2011) revealed exceptionally high intra-and interspecific distances, similar to the observations made in other Scolopendromorpha genera (see above), as well as in a recent study on German geophilomorph centipedes (Wesener et al. 2015). ...
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In order to evaluate the diversity of Central European Myriapoda species in the course of the German Barcode of Life project, 61 cytochrome c oxidase I sequences of the genus Cryptops Leach, 1815, a centipede genus of the order Scolopendromorpha, were successfully sequenced and analyzed. One sequence of Scolopendra cingulata Latreille, 1829 and one of Theatops erythrocephalus Koch, 1847 were utilized as outgroups. Instead of the expected three species (C. parisi Brolemann, 1920; C. anomalans Newport, 1844; C. hortensis (Donovan, 1810)), analyzed samples included eight to ten species. Of the eight clearly distinguishable morphospecies of Cryptops, five (C. parisi; C. croaticus Verhoeff, 1931; C. anomalans; C. umbricus Verhoeff, 1931; C. hortensis) could be tentatively determined to species level, while a further three remain undetermined (one each from Germany, Austria and Croatia, and Slovenia). Cryptops croaticus is recorded for the first time from Austria. A single specimen (previously suspected as being C. anomalans), was redetermined as C. umbricus Verhoeff, 1931, a first record for Germany. All analyzed Cryptops species are monophyletic and show large genetic distances from one another (p-distances of 13.7–22.2%). Clear barcoding gaps are present in lineages represented by >10 specimens, highlighting the usefulness of the barcoding method for evaluating species diversity in centipedes. German specimens formally assigned to C. parisi are divided into three clades differing by 8.4–11.3% from one another; their intra-lineage genetic distance is much lower at 0–1.1%. The three clades are geographically separate, indicating that they might represent distinct species. Aside from C. parisi, intraspecific distances of Cryptops spp. in Central Europe are low (<3.3%).
... among interspecific populations [77]. A clear gap was also found in the genetic distance for COX1 between and within species, as the average genetic distance within species was 6.4 and the highest was 9.1%, whereas the average genetic distance was 14.8% and the lowest was 13.5% between species [78]. Hence, our results hypothesized that T. tuberculata DL was a cryptic species of the T. tuberculata. ...
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Based on morphological characteristics to make species identification, the cryptic species of the Scutigeromorpha can be greatly underestimated. The mitochondrial genome provides a desirable tool for the biological identifications and the discovery of the cryptic species. The capacity to acquire mitochondrial genome sequences has substantially improved in recent years using next-generation sequencing (NGS) technology. On the basis of the next-generation sequencing, we obtained four complete mitochondrial genomes of Thereuonema tuberculata (Wood, 1862) from Nanyang, Henan Province (NY), Nanchang, Jiangxi Province (NC), Jinan, Shandong Province (JN), and Dali, Yunnan Province (DL) in China with GenBank numbers OK513221, OL449685, ON058988 and ON058989, respectively. The lengths of the four mitochondrial genomes ranged from 14,903 to 14,909 bp. The composition and order of genes of the four mitochondrial genomes were identical to the published mitochondrial genome of Scutigera coleoptrata (Linnaeus, 1758) (Scutigeromorpha: Scutigerdae). It was the first time that the tandem repeats in the control region were detected in Scutigeromorpha. We also calculated the corrected pairwise genetic distance of four complete mitochondrial genomes of T. tuberculata, ranging from 7.7 to 15.2%. The results showed that the T.tuberculata NC belonged to the typical sample of T. tuberculata, and T. tuberculata DL was hypothesized as a cryptic species of T. tuberculata. Meanwhile, T. tuberculata NY and T. tuberculata JN were hypothesized as potential cryptic species of T. tuberculata in this study. In both BI and ML trees, the monophyly of Scutigeromorpha, Scolopendromorpha, Geophilomorpha, and Lithobiomorpha was forcefully advocated. Moreover, Scutigeromorpha was recovered as the sister clade of (Scolopendromorpha + (Lithobiomorpha + Geophilomorpha)). Four specimens of T. tuberculata were clustered into one clade, which was the sister to the clade of S. coleoptrata.
... Morphology-based taxonomy of Scolopendra is complicated because of individual and geographical variations seen in several morphological characters that have been used by previous authors for species discrimination (e.g., Siriwut et al. 2015). Improved species discrimination has been attempted in several studies by using a combination of conventional morphology-based and modern molecular phylogenetic methods, mainly for Asian Scolopendra species (e.g., Oeyen et al. 2014;Siriwut et al. 2015b;Kang et al. 2017;Doménech et al. 2018). In the present study, we also employed an integrative approach, as described below. ...
In Japan and Taiwan, five valid species of the genus Scolopendra Linnaeus, 1758 have been described: S. morsitans Linnaeus, 1758, S. subspinipes Leach, 1816, S. mutilans Koch, 1878, S. japonica Koch, 1878, and S. multidens Newport, 1844. Recently, an undetermined species was found in the Ryukyu Archipelago and Taiwan. Using molecular phylogenetic analyses with mitochondrial COI and 16S rRNA and nuclear 28S rRNA and 18S rRNA genes as well as conventional morphological examination, we successfully discriminated this sixth species as an independent lineage from S. subspinipes, S. mutilans, and other named congeners from East and Southeast Asia. Therefore, the species was described as S. alcyona Tsukamoto & Shimano, sp. nov. Several situational evidences suggest that this species prefers streamside environments and exhibits amphibious behavior.
... However, mobile tropical genera with widely distributed species such as the large colourful fire millipedes of the genus Aphistogoniulus Silvestri, 1897, endemic to Madagascar, show larger intraspecific variation of up to 12%. Such large intraspecific distances in the COI gene seem to be the norm in European centipedes, reaching up to 15% in Geophilomorpha , as well as Scolopendromorpha (Oeyen et al 2014; and Lithobiomorpha (Decker et al. 2017), comparable to the distances observable between different populations of Polyxenus lagurus. As bisexual populations of P. lagurus have never been found in mainland Germany in previous studies (Enghoff 1978, Schömann 1956, nor have they been revealed in German specimens examined in this study, it is probable that Clade 5 ( fig. ...
The Polyxenidae in the fauna of the Crimeo-Caucasian region is represented by four species: Polyxenus lagurus (Linnaeus, 1758) (= P. lagurus caucasicus Lignau, 1924, syn. n.), Propolyxenus argentifer (Verhoeff, 1921) comb. n. (= P. trivittatus Verhoeff, 1941, = P. sokolowi Lignau, 1924, both syn. n.), a new species, Polyxenus lankaranensis sp. n., and an undescribed Polyxenus sp. The distributions of all these species in the region concerned are mapped, based on old and new records. A molecular phylogeny based on COI sequences is used to study the relationship within and among the genera Polyxenus and Propolyxenus from Western Europe to the southern Caucasus. The results highlight the presence of a number of undescribed species of Polyxenus across this region.
... For that reasons, in the last decades several studies have introduced a molecular approach as a complement to morphology-based taxonomic works (Edgecombe & Giribet 2008;Joshi & Karanth 2011, 2012Murienne et al. 2010Murienne et al. , 2011. The selected sequence of the mitochondrial cytochrome c oxidase subunit I gene (COI) has been used as a reference for taxonomic and phylogenetic studies in diverse organisms (Mengual 2008, Krishnamurthy & Francis 2012, Tyagi et al. 2017, being equally proved useful as a complement for the centipede's species delimitation (Edgecombe & Giribet 2008;Vahtera et al. 2012Vahtera et al. , 2013Oeyen et al. 2014;Siriwut et al. 2015Siriwut et al. , 2016. The analysis of the evolutionary divergence in combination with phylogenetic trees inference, using the COI gene isolated or in concatenation, has contributed for example to positioning in trees of previously undescribed species (Siriwut et al. 2015) like S. cataracta Siriwut, Edgecombe & Prahna, 2016, or to unmasking possible cryptic species resembling S. pinguis Pocock, 1891a (Siriwut et al. 2015). ...
The genus Scolopendra Linnaeus, 1758 is represented in the Philippines’ fauna by five species, two of which are endemic. Mitochondrial DNA sequences of gene cytochrome c oxidase subunit I (COI) were obtained from six Scolopendra specimens belonging to two endemic species and a new one, described here as Scolopendra paradoxa Doménech sp. nov. These sequences were analyzed together with another forty-one sequences from GenBank, including additional species of Scolopendra and a few representatives of other Scolopendridae genera. Phylogenetic trees inferred from the COI analysis using maximum likelihood and neighbor joining showed the three Philippines Scolopendra endemic species as a polyphyletic group coherent with their respective morphologies, although the position of S. spinosissima Kraepelin, 1903 varied within the obtained trees. Species delimitation based on standard external morphological characters was also concordant with the observed genetic distances, monophyly and node support, confirming S. subcrustalis Kronmüller, 2009 and S. paradoxa sp. nov. as separate species also at the molecular level, while only the position of S. spinosissima could not be properly established with any of the statistical methods used. In addition, the male genitalia of the three studied species were found to lack gonopods and a penis. Remarks on the ultimate legs prefemoral spinous formula of S. spinosissima plus a key to the species of the genus Scolopendra in the Philippines are provided.
... He also reviewed the subspecies Scolopendra subspinipes Leach, 1815 and proposed a nomen novum (Kronmüller 2012). Simaiakis et al. (2012) and Oeyen et al. (2014) studied the evolutionary history of Scolopendra cingulata Latreille, 1829 in Europe based on its morphology and molecular data, respectively. Siriwut et al. (2015) evaluated the use of morphology and molecular data on Scolopendra species delimitation from mainland Southeast Asia. ...
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Zootaxa 4425 (1): 153-164 Abstract Scolopendra arthrorhabdoides Ribaut, 1913 is redescribed based on fresh material. Its taxonomic status is evaluated and compared with Scolopendra armata Kraepelin, 1903 and Scolopendra alternans Leach, 1816. The geographical distribution of S. arthrorhabdoides is also revised. Scolopendra armata is reported from Colombia for the first time.
... On the other hand, the distance analysis can offer reference points in timing slowly evolving relicts, where molecular taxonomy cannot provide convincing results. Oeyen and his colleagues tested the "speciation by distance" (both in space and time) phylogeographic hypothesis in the Pannonian region by combining topographic and molecular genetic approaches on the Mediterranean Banded Centipede (Scolopendra cingulata Latreille) [63]. The authors studied the rediscovered Austrian and the already known Hungarian and Romanian occurrences as examples isolated by great distance, genetically comparing them to the populations of the continuous Mediterranean coastal areas. ...
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The identification of climatic relicts is seldom straightforward. These species are threatened owing to current climatic trends, which underlines the importance of carrying out ecological and biogeographic investigations of them. Here we introduce a novel approach to improve the identification of climatic relicts. We are focusing on thermophilic relict plants of the Pannonian biogeographic region from the Holocene Thermal Maximum (HTM). We argue that a minimal mean annual temperature difference (MATD) of the HTM compared to the recent climate allowed a continuous northward expansion for the taxa investigated. We measured latitudinal distances between the recent occurrences of relicts and those of the main distribution found further south. Regarding estimates for MATD (1.0–2.5 °C), we only consider species with a distribution which has a 150–350 km North-South gap, since a latitudinally directed distance can be translated into temperature, showing a poleward cooling trend. Of the 15 selected species, 12 were recorded with values of 1.0–1.7 °C MATD, and three with values of 1.8–2.5 °C, some of which are presumably interglacial species. Woody species are over-represented among them (four species), using the Hungarian flora as a reference. The proposed method allows the prediction of potential climate-related changes in the future distribution of species, constrained by the topographic features of their habitats.
... Karakteriše je pljosnato telo dužine do 17cm, koje može pokazivati značajne varijacije u boji, kao i prvi par nogu koji je modifikovan u klješta na kojima se nalaze izvodni putevi otrovnih žlezda (3) . Scolopendra cingulata je izuzetno agresivna vrsta, hrani se insektima i manjim gmizavcima, ali ukoliko je u prilici, napada i višestruko veće organizme (4) . ...
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Scolopendra cingulata (Laterille, 1829) was noticed sixty years ago for the first time in one location in Serbia and since then data about it are scarce. It represents the biggest centipede in Europe and only one that can seriously harm humans. Scolopendrism in Republic of Serbia is a true rarity and until now there has been no case report of harming human by centipede Scolopendra cingulata (Laterille, 1829). Patient aged 85, was stung on his right leg toe by unknown centipede in the toilet of his home. The centipede was afterwards identified as Scolopendra cingulata. As a chief complaint patient describes intense pain at the sting site and burning feeling that spreads to the hip joint. The objective finding revealed a slight redness of the affected area. He was ordinated 20mg chloropyramine (Synopen), 2x4mg dexamethasone and 40mg methylprednisolone i.v. 24 hours after centipede attack pain was completely gone and the patient could walk freely. Although there is a possibility that a species located in our country's territory exhibits phenotypes and genotype differences in relation to the more widely described Mediterranean type, each human bitten by i.e. that come into contact with Scolopendra cingulata venom should be taken seriously in order to facilitate the patient's symptoms, manage constant monitoring and prevent the development of isolated or associated complications with potential comorbidities of the patient.
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The presence of an isolated population of Scolopendra cingulata in NW Italy is here reported, together with the vegetational characteristics of the biotope. These data further underline the particularity and the importance of the conservation of this site.
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ALESSANDRO MINELLI Istituto di Zoologia de11’Um'versit'21 di Padova Note critiche sui Chilopodi della Sardegna La rnaggior parte -dei dati finora pubblioati sui Chilopodi della Sardegna e contenuta in soli quattro lavori, due antichi (Fan- zago, 1881; Silvestri, 1897) ie du-e modern-i (Eason, 1980; Minelli, 1983); altre informazioni, compresa la descrizione di alcune specie e sottospecie nuove, sono sparse in un’altra ventina di lavori, tutti elencati in bibliografia. A110 stato presente delle nostre conoscenze, ritengo si possa considerate accertata Ia presenza in Sardegna di 43 specie di Chi— lopodi. Alle 37 di cui mi sono occupato nella nota precedente (Minelli, 1983) vannoinfatti aggiunti: -— Litbobius doderoi Si1V., specie cavernicola di cui ho esa- minato la serie tipica; —— Plzztoimmz zzuierleinii Cavanna, Lit/aolaiux calccmztus C. Koch e L. dczbli Verh., 1e cui citazioni per la Sardegna sono attendibili, trattandosi di specie di facile i-dentificazione e non ponendo, 1a loro presenza in Sardegna, particolari problemi zoogeognafici; —— Lit/aobius cerii Verh., con cui identifico, anche se con qualche esitazione, una specie recentemente descritta come nuova da Restivo de Miranda (197813): Lit/aolaim molop/mi (la sinonirnia e discussa pifi avanti). — Lit/aobius agilis sandy: Manfr., la cui precisa didentitet e i cui rapporti con L. agilis C. Koch mi sono ignoti, ma che non e identifioabile con a1c1in’a1tra specie nota per la Sardegna. Prima di passare ad elencare 1e specie Ia cui presenza in Sat- degna e accertata e a discuterne il significato biogeografico, e_;.«ne-
The species of the genus Scolopendra Linnaeus, 1758 are very widely distributed in the Mediterranean region. Current knowledge is summarized, with references to material derived mainly from the well-explored islands of the Aegean Archipelago, several localities in continental Greece and old bibliographic reports. We suggest that Scolopendra species represent examples of both paradigms of historical biogeography, namely vicariance and dispersal. We propose that the dispersal routes of Scolopendra species in Greece were mainly influenced by late Miocene and upper Pleistocene palaeogeography. The formation of the Mid-Aegean trench (c. 12 – 9 Mya) considered as a remarkable geographical barrier between the Anatolian peninsula and mainland Greece, prevented the entry of certain Scolopendra species westwards. In total, five Scolopendra species have been recorded from mainland and insular Greece. A vicariance event that occurred in the area more than 17 Mya, when the Aegean region was part of a united landmass, better explains the biogeographical history of S. canidens. Cyclades harbours remnants of the ancient populations of S. canidens, whereas during the late Pleistocene (c. 400.000 21.000 ya) S. canidens was isolated in Dodecanese. S. cretica is the only endemic, being distributed in Crete and its adjacent islets. S. clavipes in E-Mediterranean and S. dalmatica in W-Mediterranean evolved from ancient canidens populations. S. cingulata entered central and southern Europe from the east (c. 20 – 11 Mya), while the formation of the Mid-Aegean trench (c. 12 – 9 Mya) prevented its entry in Crete.
A new method called the neighbor-joining method is proposed for reconstructing phylogenetic trees from evolutionary distance data. The principle of this method is to find pairs of operational taxonomic units (OTUs [= neighbors]) that minimize the total branch length at each stage of clustering of OTUs starting with a starlike tree. The branch lengths as well as the topology of a parsimonious tree can quickly be obtained by using this method. Using computer simulation, we studied the efficiency of this method in obtaining the correct unrooted tree in comparison with that of five other tree-making methods: the unweighted pair group method of analysis, Farris's method, Sattath and Tversky's method, Li's method, and Tateno et al.'s modified Farris method. The new, neighbor-joining method and Sattath and Tversky's method are shown to be generally better than the other methods.
During the last ten years, the use of molecular markers, revealing polymorphism at the DNA level, has been playing an increasing part in animal genetics studies. Amongst others, the microsatellite DNA marker has been the most widely used, due to its easy use by simple PCR, followed by a denaturing gel electrophoresis for allele size determination, and to the high degree of information provided by its large number of alleles per locus. Despite this, a new marker type, named SNP, for Single Nucleotide Polymorphism, is now on the scene and has gained high popularity, even though it is only a bi-allelic type of marker. In this review, we will discuss the reasons for this apparent step backwards, and the pertinence of the use of SNPs in animal genetics, in comparison with other marker types.