www. europeanj ournaloftaxonomy. eu
2016 • Pimvichai P. et al.
http://dx.doi.org/10.5852/ejt.2016.199
This work is licensed under a Creative Commons Attribution 3.0 License.
Research article
urn:lsid:zoobank.org:pub:EE35A675-06C7-4FF9-9EE0-0854D95036C4
A revision of the Thyropygus allevatus group. Part V: Nine new species
of the extended opinatus subgroup, based on morphological and DNA
sequence data (Diplopoda: Spirostreptida: Harpagophoridae)
Piyatida PIMVICHAI Henrik ENGHOFF 2 , Somsak PANHA 3 & Thierry BACKEFJAU 4
1 Department of Biology, Faculty of Science, Mahasarakham University,
Kantharawichai District, Maha Sarakham 44150, Thailand.
2 Natural History Museum of Denmark, University of Copenhagen,
Universitetsparlcen 15, DK-2100 Copenhagen 0, De nm ark
3 Animal Systematics Research Unit, Department of Biology, Faculty of Science,
Chulalonglcorn University, Bangkok 10330, Thailand.
4 University of Antwerp, Evolutionary Ecology Group, Groenenborgerlaan 171,
B-2020 Antwerp, Belgium and Royal Belgian Institute of Natural Sciences,
OD Taxonomy and Phylogeny, and JEMU, Vautierstraat 29, B-1000 Brussels, Belgium.
* Corresponding author: [email protected]
2 Email: henghoff@snm ku.dk
3 Email: [email protected]
4 Email: [email protected]
1 urn:lsid:zoobank.org:author:4D882A89-E0Cl-4F78-93E7-FAlB5A2650F2
2 urn:lsid:zoobank.org:author:FB09A817-000D-43C3-BCC4-2BClE5373635
3 urn:lsid:zoobank.org:author:AC935098-D901-4F35-A414-4B0D4FE44E79
4 urn:lsid:zoobank.org:author:E6F42575-36AE-4AD7-98C6-D083EF052568
Abstract. The Thyropygus opinatus subgroup (Diplopoda: Harpagophoridae) of the T. allevatus group
in Thailand is revised. Based on a phylogenetic analysis of mtDNA sequence data, it is merged with
the T. bifurcus subgroup to form an extended T. opinatus subgroup. Nine new species are described:
Thyropygus cimi sp. nov. and T. forceps sp. nov. from Nakhonsrithammarat Province, T. culter sp. nov.,
T. planispina sp. nov., T. undulatus sp. nov. and T. ursus sp. nov. from Krabi Province, T. mesocristatus
sp. nov. from Songkhla Province, T. navychula sp. nov. from Phang-Nga Province and T. sutchariti
sp. nov. from Phetchaburi Province.
Keywords. Millipedes, DNA barcoding, phylogeny, new species, Thailand.
Pimvichai R, Enghoff H., Panha S. & Backeljau T. 2016. A revision of the Thyropygus allevatus group. Part V:
Nine new species of the extended opinatus subgroup, based on morphological and DNA sequence data (Diplopoda:
Spirostreptida: Harpagophoridae). European Journal of Taxonomy 199: 1-37. http://dx.doi.org/10.5852/
eit.2016.199
1
European Journal of Taxonomy 199 : 1-37 ( 2016 )
Introduction
The millipede genus Thyropygus Pocock, 1894, is the most species-rich genus of the family
Harpagophoridae (subfamily Harpagophorinae) in Southeast Asia. Currently, it includes 56 named
species of which 35 occur in Thailand (Enghoff 2005; Jeelcel 2006; Pimvichai et al. 2009a, b; 2011a, b).
The genus is widely distributed in Southeast Asia and is particularly diverse in Thailand. In a series of
previous papers four subgroups of the large Thyropygus allevatus group were revised: the T. opinatus
subgroup (Pimvichai et al. 2009a), the T. bifurcus subgroup (Pimvichai et al. 2009b), the T. induratus
subgroup (Pimvichai et al. 2011a), and the T. cuisinieri subgroup (Pimvichai et al. 2011b). Yet, several
species of the T. allevatus group still have not been assigned to a subgroup.
Although the four currently recognized subgroups of the T. allevatus group appear morphologically
distinct, Pimvichai et al. (2014) observed that the T. opinatus and T. bifurcus subgroups did not form
separate clades in a mtDNA phylogeny, but instead formed a single mixed clade. This suggested that
the two subgroups should not be kept separate, which would also better reflect their morphological
similarity (Pimvichai et al. 2009b).
The mitochondrial cytochrome c oxidase subunit I (COI) is widely used for DNA-based species
identification (DNA barcoding) of closely related species (Hebert et al. 2003), while mitochondrial
16S rRNA sequences are often used as an additional marker in diplopod taxonomy and phylogeny
(Pimvichai et al. 2014). Hence, in the present study, COI and 16S rRNA sequence data are used to
(1) delimit species of the T. opinatus and T. bifurcus subgroups, and (2) assess the monophyly of both
these subgroups. As a result we describe nine new species and suggest to abandon the concept of a
separate T. bifurcus subgroup. Instead, we propose to merge the T. bifurcus and T. opinatus subgroups
into an extended T. opinatus subgroup.
Material and methods
Fresh specimens were hand-collected and preserved partly in 70% ethanol, partly in a freezer at -20 °C
for subsequent DNA studies. Specimens were examined from the following collections:
CUMZ = Museum of Zoology, Chulalongkorn University, Bangkok, Thailand
NHMW = Naturhistorisches Museum, Vienna, Austria
ZMUC = Zoological Museum, Natural History Museum of Denmark, University of Copenhagen
Partial COI and 16S rRNA gene sequences were used to construct a phylogenetic tree of Thyropygus
species. COI was also used to evaluate genetic divergences between species. The DNA sequence data
were extracted from Pimvichai et al. (2014) and supplemented with sequences of seven new samples.
The procedures for extracting and analyzing COI and 16S rRNA sequences followed Pimvichai et al.
(2014). The COI fragment was amplified using the primers ECO-1490 and HCO-2198 (Folmer et al.
1994), while the 16S rRNA DNA fragment was amplified with the universal primers 16Sar and 16Sbr
(Kessing et al. 1989). The new sequences are deposited in GenBank under the accession numbers
KU306518-KU306531. Collection localities and accession codes for each nominal species are shown
in Table 1.
The COI and 16S rRNA data involve 33 specimens, representing 29 named species. The genus
Anurostreptus Attems, 1914 belongs to another harpagophorid subfamily (Rhynchoproctinae), and
therefore Anurostreptus barthelemyae Demange, 1961, and A. sculptus Demange, 1961 were used as
outgroup taxa.
2
PIMVICHAIP. et al ., New species of Thyropygus millipedes
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European Journal of Taxonomy 199: 1-37 (2016)
Alignment and phylogenetic analysis
The resulting forward and reverse sequences were assembled using CodonCode Aligner version
4.0.4 (CodonCode Corporation) and checked for errors/ambiguities. The partial nucleotide sequences
(COI, 16S rRNA) were analyzed with the Basic Tocal Alignment Search Tool (BTAST) provided by
NCBI to compare them with known reference sequences in GenBank. DNA sequences were aligned
using MUSCTE (Edgar 2004). The two original mtDNA alignments consisted of 660 bp (COI) and
514 bp (16S rRNA). Yet, 27 bp, including all the gaps, were excluded from the 16S rRNA because
of alignment ambiguities, resulting in a final 16S rRNA alignment of 487 bp. The sequences were
checked for ambiguous nucleotide sites, saturation and phylogenetic signal using DAMBE version
5.2.65 (Xia 2013). MEGA version 6 (Tamura et al. 2013) was used to (1) perform Tajima’s D test for
selective neutrality of mutations (D-test statistic), (2) calculate uncorrected pairwise p-distances among
sequences, (3) translate COI protein coding sequences into amino acids, (4) check for stop codons, and
(5) evaluate transition/transversion rates for COI. Substitution models were inferred independently for
each of the gene partitions using jModelTest version 2.1.7 (Darriba et al. 2012) applying Akaike (1974)
weights as selection criterion.
Phylogenetic trees were constructed using maximum likelihood (ML) and Bayesian inference (BI). The
shape parameter of the gamma distribution, based on 16 rate categories, was estimated using likelihood
analysis. ML trees were inferred with RAxML version 8.0.0 (Stamatakis 2014) through the CIPRES
Science Gateway (Miller et al. 2010) using a GTR+G substitution model and 1000 bootstrap replicates
to assess branch support. The concatenated sequence alignment was partitioned by gene, allowing
different evolution rates for COI and 16S rRNA.
The BI analysis was run for 5 million generations (heating parameter = 0.08), sampling every 1000
generations. Convergence was confirmed by verifying that the standard deviations of split frequencies
were below 0.01. Then the first 2500 trees were discarded as burn-in, so that the final consensus tree
was built using the last 7502 trees. The optimal parameters were assessed by calculating the Bayes
factor using Tracer version 1.6.0 (Rambaut et al. 2014). Support for nodes was defined as posterior
probabilities.
Species descriptions
Drawings were made using a stereo microscope. We have mainly focused on adult males, although adult
females and a few juveniles were also available. For the gonopods we used the terminology of Pimvichai
et al. (2009a: 21, figs 1-3) with the following abbreviations (terms not used in Pimvichai et al. 2009a,
b in bold):
ac = anterior coxal fold: the main part of gonopod in anterior view; confusingly called posterior
coxal fold by Demange (1961) and Hoffman (1975)
aip = additional spine-like process: between lateral and mesal processes of anterior coxal fold
alp = lateral process of anterior coxal fold: the distolateral part of the anterior coxal fold
amp = mesal process of anterior coxal fold: an additional projection on the anterior coxal fold,
protruding from its mesal margin
bp = blepharochaete (pi. -ae): the normal form of apical setae, long, slender, stiffened, and usually
pigmented, somewhat reminiscent of the mammalian eyelash (Hoffman 1975)
cr = longitudinal crest in gutter of palette: a crest which runs along the middle of the gutter near the
tip of the palette
fe = femoral spine (also fe 1 and fe 2)\ a usually long, curved spine on the telopodite, originating
slightly distal to the point where the telopodite emerges from the coxa
Ic = longitudinal crest: a strong longitudinal crest at the mesal margin of amp in posterior view
ll = lamellar lobe: a small, slightly folded lobe at the basis of the apical part of the telopodite
pa = palette: the distalmost lobe of the apical part, carrying the row of blepharochaetae
6
PIMVICHAIP. et al ., New species of Thyropygus millipedes
pc = posterior coxal fold: the main part of gonopod in posterior view, usually shorter than ac and
forming shelf for accommodation of telopodite shaft
pip = lateral process of posterior coxal fold: the lateral part of the posterior coxal fold, normally
digitiform
pmp = mesal process of posterior coxal fold: the mesal part of the posterior coxal fold, normally
forming a shelf for accommodation of telopodite shaft
px = paracoxite: the basal, lateral part of the posterior coxal fold
si = spatulate lobe: a distinct distal, separate lobe at the apical part, spatulate, sometimes with a
distal spine-like process
sis = slender long spine: an additional slender long spine (much longer than ss ) at the base of the
apical part of telopodite in posterior view
ss = small spine: an additional small spine at the base of the apical part of telopodite in posterior
view
st = sternum: a small, usually triangular sclerite between the basal parts of the anterior coxal folds
ti = tibial spine: a usually long spine on the telopodite, originating distal to the femoral spine, at the
basis of the apical part of the telopodite, usually curved in the opposite direction of the femoral
spine, the two together forming a circle
Apical part: the part of the telopodite distal to the tibial spine
Shelf: the distal surface of the posterior coxal fold
Results
DNA sequence data
The COI dataset included 660 bp, while the 16S rRNA dataset included 487 bp. The concatenated
dataset therefore comprised 1147 bp.
The aligned COI gene fragment (660 bp) had nucleotide frequencies of 0.287, 0.226, 0.161 and 0.326
for A, C, G and T, respectively, (37.8% GC content). The uncorrected p-distance between the taxa
ranged from 0.02 to 0.21 (Table 2).
The aligned 16S rRNA gene fragment (487 bp), had nucleotide frequencies of 0.324, 0.100, 0.210 and
0.366 for A, C, G and T, respectively (29.9% GC content). The uncorrected p-distance between the taxa
ranged from 0.00 to 0.15 (Appendix I).
The concatenated data set (1147 bp) had nucleotide frequencies of 0.303, 0.172, 0.182 and 0.343 for
A, C, G and T, respectively (35.3% GC content). The uncorrected p-distance between the taxa ranged
from 0.01 to 0.18. The estimated value of the shape parameter, as evaluated by MEGA version 6 with
the substitution pattern and rates fitted by the GTR+G model (Kumar & Nei 2000), for the discrete
Gamma Distributions (Gu et al. 1995) were 0.1542, 0.1584 and 0.1424 for the COI fragment, 16S rRNA
(unambiguously aligned sequences) and the concatenated data set, respectively.
D values for selective neutrality were 1.91, 0.63 and 1.45 for COI, 16S and the combined sequences,
respectively. The COI transition/transversion ratio was 2.24.
Phytogeny
For ML analyses, we consider clades with bootstrap values of 70% and above to be well-supported (Hillis
& Bull 1993), and below as not significantly supported; thus, we refer to clades with bootstrap values
between 70% and 79% as moderately supported, between 80% and 89% as strongly supported, and 90%
and above as very strongly supported. For BI analyses, we regard clades with posterior probabilities of
0.95 and above to be strongly supported and below as not significantly supported.
7
European Journal of Taxonomy 199: 1-37 (2016)
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31 T. uncinatus Rcb 0.16 0.16 0.13 0.13 0.14 0.13 0.14 0.16 0.15 0.16 0.15 0.15 0.14 0.15 0.16 0.13 0.13 0.15 0.15 0.13 0.16 0.15 0.15 0.16 0.15 0.12 0.13 0.15 0.15 0.00
32 T. undulatus sp. nov. Pb2 0.18 0.20 0.15 0.15 0.12 0.15 0.17 0.11 0.13 0.03 0.13 0.13 0.05 0.17 0.17 0.17 0.17 0.16 0.16 0.14 0.13 0.15 0.12 0.02 0.12 0.17 0.17 0.13 0.16 0.16 0.16
33 T. ursus sp. nov. LTK 0.20 0.19 0,16 0,16 0.05 0,13 0,13 0,11 0.05 0,12 0.08 0.04 0,11 0,15 0,13 0.14 0,14 0,12 0.15 0,11 0.07 0.11 0,12 0,12 0.02 0.14 0,14 0.05 0.14 0,15 0,15 0.12
PIMVICHAIP. et al. , New species of Thyropygus millipedes
The ML and BI phylogenetic trees reconstructed by separate and combined analyses of the partial COI
and 16S rRNA were largely congruent (by visual inspection of the branch pattern). Indeed, the COI
and concatenated COI+16S rRNA trees were topologically identical, whereas the 16S rRNA tree only
showed topological differences for unresolved or unsupported nodes. The single gene trees (COI and
16S rRNA) are presented in Appendices II and III. Thus, we have selected the combined COI plus 16S
rRNA phytogeny derived from the ML analysis for further discussion (Fig. 1). The best-fit substitution
models for the ML tree under the Alcailce information criterion (Akaike 1974) was the TIM1 + I + G
(-InL = 5531.3288, gamma shape = 0.7890) and TIM3 +1 + G (-InL = 2557.2902, gamma shape = 0.4540)
model for COI and 16S rRNA, respectively. The Thyropygus clade (Clade 1, 27 included species) is
strongly supported by 100% for ML bootstrap replicates, and a BI posterior probability of 1.00.
Clade 1A1, Thyropygus allevatus is very strongly supported (100% for ML bootstrap replicates and a
BI posterior probability of 1.00).
100 / 1.00
—Anurostreptus barthelemyae Tib
■Anurostreptus sculptus PI
100 / 1.00
100 / 1.00
78/1.00
#/0.81
65/0.86
Thyropygus allevatus Bb
Thyropygus allevatus Bk
100/1.00 i - Thyropygus foliaceus Kj
I- Thyropygus jarukchusri Np
81/0.991- Thyropygus bispinus Tm
T
1A1
1
T
1A2
89/1.00
#/-
#/-
59/0.93
#/-
77/0.88
91/0.99
69/0.78
53/0.88
88/1.00
55/0.77
98/1.00
uT=
!_I98/1.C
A_[
Thyropygus inflexus Kk
Thyropygus brachyacanthus Sk
Thyropygus enghoffi Kl
Thyropygus cris tag alii Yn
Thyropygus undulatus sp. nov. Pb 2
Thyropygus planispina sp. nov. Tsk 2
Thyropygus cutter sp. nov. Nr
Thyropygus bearti Ks
- Thyropygus mesocristatus sp. nov. Ss
I —Thyropygus cimi sp. nov. Nws 1_2
J j- Thyropygus quadricuspis Hy
Jin —Thyropygus ursus sp. nov. LTK
11 — Thyropygus demangei Cm
I- Thyropygus richardhoffmaniTCH
- Thyropygus forceps sp. nov. Nws 2
143
81/0.98
100/1.00
Thyropygus bispinispatula Kb
- Thyropygus opinatus Arw
— Thyropygus navychula sp. nov. KSR
Thyropygus loxia Ty
- Thyropygus sutchariti sp. nov. KCP
#/0.70
100 / 1.00
100 / 1.00
I Thyropygus uncinatus Kdb
1 -Thyropygus uncinatus Rcb
Thyropygus resimus Kc
Thyropygus resimus Ksp
I— Thyropygus induratus Nru
I Thu
1A4
'Thyropygus induratus Tt
0.06
Fig. 1. Phylogenetic relationships of Thyropygus species based on maximum likelihood analysis (ML)
and Bayesian Inference (BI) of 1147 bp of concatenated gene fragments of COI (660 bp) and 16S
rRNA (487 bp). Numbers at nodes indicate branch support based on bootstrapping (ML) / posterior
probability (BI). Scale bar = 0.06 substitutions/site. # indicates branches which received < 50% ML
bootstrap support, - indicates non-supported branches by posterior probability. Clade memberships
and designations are shown as vertical bars; 1A1 = T. allevatus , 1A2 = cuisinieri subgroup, 1A3 =
opinatus subgroup and 1A4 = induratus subgroup. The coloured area marks the T. opinatus subgroup.
Abbreviations after species names refer to locality names as shown in Table 1.
9
European Journal of Taxonomy 199: 1-37 (2016)
Clade 1A2, T. cuisinieri subgroup (here represented by foliaceus and jarukchusri ) is very strongly
supported (100% for ML bootstrap replicates and a BI posterior probability of 1.00).
Clade 1A3 (coloured highlight), is strongly supported by 89% ML bootstrap replicates and strongly
supported by a BI posterior probability of 1.00, and is a monophyletic group consisting of a mixture of
species from the T. opinatus and T. bifurcus subgroups (Pimvichai et al. 2009a, b), including the nine
new species described in this study. Thus, we treat all species of these subgroups as a single subgroup
viz., the extended T. opinatus subgroup (henceforward simply “the T. opinatus subgroup”).
Clade 1A4, the T. induratus subgroup ( induratus, resimus and uncinatus) is not significantly supported
(65% for ML bootstrap replicates and a BI posterior probability of 0.86).
Taxonomy
The Thyropygus opinatus subgroup (including the T. bifurcus subgroup of Pimvichai et al. 2009b)
This is a subgroup of the T. allevatus group, characterized by an additional projection on the anterior
coxal fold (amp). Included species:
T. beard Pimvichai et al., 2009a
T. bifurcus (Demange, 1986)
T. bispinispatula Pimvichai et al., 2009a
T. bispinus Pimvichai et al., 2009a
T. brachyacanthus Pimvichai et al., 2009a
T. casjeekeli Pimvichai et al., 2009b
T. chelatus Pimvichai et al., 2009a
T. cristagalli Pimvichai et al., 2009a
T. demangei Pimvichai et al, 2009b
T. enghoffi (Demange, 1989)
T. erectus Pimvichai et al, 2009a
T. floweri (Demange, 1961)
T. implicatus (Demange, 1961)
T. inflexus (Demange, 1989)
T. loxia Pimvichai et al., 2009a
T. opinatus (Karsch, 1881)
T. quadricuspis Pimvichai et al., 2009b
T. richardhoffmani Pimvichai et al., 2009b
T. cimi sp. nov.
T. culter sp. nov.
T. forceps sp. nov.
T. mesocristatus sp. nov.
T. navychula sp. nov.
T. planispina sp. nov.
T. sutcharid sp. nov.
T. undulatus sp. nov.
T. ursus sp. nov.
In addition to the diagnostic character combination given by Pimvichai et al. (2009a, b), all species in
the opinatus subgroup share common characters of head, antennae, mandibles, gnathochilarium, collum,
epiproct, paraprocts, hypoproct and first pair of male legs. We therefore give a general description of
males of the opinatus subgroup, so that subsequent species descriptions (which are mainly based on
males) can be kept brief.
10
PIMVICHAIP. et al ., New species of Thyropygus millipedes
General description of males of the T. opinatus subgroup
Head smooth, 5-8 supralabral setae, eyes moderate in size, ocelli in 7-8 horizontal rows consisting of
11-14, 10-13, 8-11, 7-9, 5-8, 3-6, 1-4 and 1-3 ocelli, total: 46-68.
Gnathochilarium: mentum smooth, at most with a few small setae distally and a large, horseshoe-shaped
ridge opening distally. Stipites densely covered with spine-lilce setae, except for an irregular oblique
band from c. middle of lateral margin to border of lamella lingualis; long setae present on the distolateral
part; males (not females) distally with a small sclerotised ‘island’ with 1-3 spine-like setae in middle of
an ovoid, poorly sclerotized, hairless area. Lamellae linguales with three long, apical setae and a number
of short, basal, spine-like setae.
Sterna smooth, stigmatal grooves distinct, long. Legs relatively long; tip of postfemora, tibiae and
tarsi visible from above when legs are extended laterad; coxae of anterior and posterior pair markedly
dissimilar, posterior coxae humped; coxae with 2-8 setae, prefemora with 3-6 setae, femora with 2-5
setae in a median row and 2 larger, subapical setae; tarsi typically with 6-8 irregularly placed ventral
setae and 1-2 large dorsal, apical spurs. Postfemora and tibiae without setae.
Ventral pads on postfemora and tibiae on all legs, except first three pairs.
Gonopods: sternum ( st ) triangular. Anterior coxal fold (ac) basally slender, becoming broader towards
tip, lateral margins diverging; distally with two processes: a lateral process (alp) and a usually smaller
mesal process (< amp ); the shape of these processes is species-specific. Posterior coxal (pc) fold much
lower than anterior coxal fold, basally with moderately high lateral paracoxites (px ), distally variously
modified but always with a smooth area over which the telopodite shaft can slide. Telopodite with a single
or double, well-developed femoral spine (fe) and a long, slender, curved tibial spine (ti). Many species
with a characteristic spatulate lobe (si) originating under the base of the tibial spine; lobe sometimes
distally rounded, spoon-like, sometimes ending in a large, stout spine. Other species instead with a small
and slightly folded lateral lamella (11). Apical palette (pa) simple, forming a broad gutter, sometimes
with a longitudinal crest in the concavity; apically with a row of 7-13 brownish blepharochaetae (bp).
Key to species of the T. opinatus subgroup (partly extracted and adapted from Pimvichai et al.
2009a, b)
1. Apical part of telopodite with spatulate lobe (si) .2
- Apical part of telopodite with lateral lamella (//).20
2. Spatulate lobe (si) distally drawn out into one (rarely two) sharp dark brown spine(s) .3
- Spatulate lobe (si) distally expanded and/or rounded, spoon-like, without a spine .9
3. Spatulate lobe (si) terminating in two sharp brown spines, the outer spine slightly smaller and
shorter than the inner one; lateral process of anterior coxal fold (alp) slender, slightly curving
mesad; mesal process of anterior coxal fold (amp) almost as long as alp , flattened.
. T. bispinispatula Pimvichai et al ., 2009a
- Spatulate lobe (si) terminating in a single sharp dark brown spine .4
4. Telopodite without a lobe distal to femoral spine (fe); lateral process of anterior coxal fold (alp)
long, slender, regularly curved, tip close to tip of opposite alp , the two together forming a circle;
mesal process of anterior coxal fold (amp) straight, shorter than alp; fe directed distad, pointed
. T. erectus Pimvichai et al ., 2009a
- Telopodite distally to fe with a large, round lobe (lo), projecting distolaterally .5
11
European Journal of Taxonomy 199: 1-37 (2016)
5. Lateral process of anterior coxal fold (alp) very slender, regularly curved .6
- Lateral process of anterior coxal fold (alp) different, broader and/or with several apical denticles ...8
6 . Mesal margin of lateral process of anterior coxal fold (alp) with fine serrations (Fig. 6A); mesal
process of anterior coxal fold (amp) almost as long as alp , broadly expanded, apically sharp,
straight distad, mesal margin forming a strong longitudinal crest (Ic) in posterior view (Fig. 6B)
. T. navychula sp. nov.
- Mesal margin of lateral process of anterior coxal fold (alp) without serrations, tip of lateral
process close to tip of the opposite side, the two together forming a circle .7
7. Mesal process of posterior coxal fold (pmp) strongly developed along anterior-posterior axis .
. T. floweri (Demange, 1961)
- Mesal process of posterior coxal fold (pmp) slender, directed distolaterad (Fig. 4C) .
. T. forceps sp. nov.
8 . Lateral process of anterior coxal fold (alp) broad, apically gradually narrowed; mesal process
of anterior coxal fold (amp) almost as long as alp , slender, straight, terminally slightly curved,
pointed. T. opinatus (Karsch, 1881)
- Lateral process of anterior coxal fold (alp) apically bent abruptly mesad, tip with serrate
margins; mesal process of anterior coxal fold (amp) much shorter than alp , directed meso-distad,
simple, pointed; mesal process of posterior coxal fold (pmp) strongly developed along anterior-
posterior axis . T. implicatus (Demange, 1961)
9. Telopodite with a single femoral spine.10
- Telopodite with a double femoral spine.17
10. Lateral process of anterior coxal fold (alp) apically abruptly truncate .
. T. bearti Pimvichai et al., 2009a
- Lateral process of anterior coxal fold (alp) apically pointed.11
11. Mesal process of anterior coxal fold (amp) very small; telopodite distally to fe with a small
round lobe (lo) projecting distolaterally. T. loxia Pimvichai et al., 2009a
- Mesal process of anterior coxal fold (amp) relatively long .12
12. Mesal process of anterior coxal fold (amp) shorter than lateral process (alp) .13
- Mesal process of anterior coxal fold (amp) as long as lateral process (alp) .14
13. Mesal process of anterior coxal fold (amp) directed obliquely disto-mesad, slender, straight.
. T. chelatus Pimvichai et al., 2009a
- Mesal process of anterior coxal fold (amp) directed distad, thicker, slightly sigmoid.
. T. br achy acanthus Pimvichai et al., 2009a
14. Mesal process of anterior coxal fold (amp) directed obliquely disto-mesad, tip overlapping
tip of opposite amp (Fig. 8A); lateral process of posterior coxal fold (pip) a massive, broad
lobe, projecting laterad (Fig. 8B) . T. sutchariti sp. nov.
- Mesal process of anterior coxal fold (amp) directed distad.15
15. Lateral process of anterior coxal fold (alp) apically without a crest; telopodite distally with a
rounded lobe (lo); margins of spatulate lobe (si) terminally meeting in a distinct angle .
. T. bispinus Pimvichai et al., 2009a
- Lateral process of anterior coxal fold (alp) apically with a crest .16
12
PIMVICHAIP. et al., New species of Thyropygus millipedes
16. Mesal process of anterior coxal fold (amp) apically irregularly tuberculate; telopodite distally
without a rounded lobe ( lo ). T. inflexus (Demange, 1989)
- Mesal process of anterior coxal fold (amp) slender, straight, its tip pointed, its mesal margin
forming a strong longitudinal crest ( Ic) in posterior view (Fig. 5B). T. mesocristatus sp. nov.
17. Anterior coxal fold (ac) with an additional spine-like process (aip) between lateral process of
anterior coxal fold (alp) and mesal process of anterior coxal fold ( amp); alp broad, mesal margin
concave, tip with serrate margins, cockscomb-like; amp much shorter than alp , directed meso-distad,
simple, pointed; both femoral spines (fe) slender, long. T. cristagalli Pimvichai et al., 2009a
- Anterior coxal fold (ac) without an additional spine-like process (aip) between alp and amp ...18
18. Lateral process of anterior coxal fold (alp) apically without a crest, flattened, slightly curved, its latero-
distal margin coarsely dentate, terminating in a short, sharp, pointed spine (Fig. 3A); mesal process
(amp) much shorter than alp, directed distad, tip curving mesad, pointed; both femoral spines (fe 1,
fe 2) long, curving backward; tibial spine (ti) long, not curving in horizontal plane (Fig. 3C).
. T. culter sp. nov.
- Lateral process of anterior coxal fold (alp) apically with a crest extending caudad .19
19. Lateral process of anterior coxal fold (alp) flattened, curving mesad, laterodistal margin coarsely
dentate, terminating in a short spine, tip curving against the tip of opposite side (Fig. 9A);
mesal process of anterior coxal fold (amp) much shorter than alp, slender, curving mesad; both
femoral spines {fe 1, fe 2) broad, long; tibial spine (ti) long, curving in horizontal plane, not
ending in a sharp spine (Fig. 9D). T. undulatus sp. nov.
- Lateral process (alp) regularly curved, terminating in a sharp, slightly upward-pointing spine
(Fig. 7A); mesal process (amp) slightly shorter than alp, flattend, straight, directed distad; tibial
spine (ti) flattend, short, curving mesad (Fig. 7D). T. planispina sp. nov.
20. Telopodite with a single femoral spine .21
- Telopodite with a double femoral spine .23
21. Lateral process of anterior coxal fold (alp) without an apical crest; mesal process of anterior
coxal fold (amp) shorter than and as broad as alp, directed distad; femoral spine (fe) very
long and slender . T. casjeekeli Pimvichai et al., 2009b
- Lateral process of anterior coxal fold (alp) with a sharp crest on the posterior surface near the tip .. .22
22. Lateral process of anterior coxal fold (alp) flattened, slightly curved, inflexed; femoral spine (fe)
very long, slender, with an additional lamella at base ...I quadricuspis Pimvichai et al., 2009b
- Lateral process of anterior coxal fold (alp) regularly curved, basally broad, gradually tapering
towards end and ending in sharp point (Fig. 2A); femoral spine (fe) very long, slender, without an
additional lamella at base (Fig. 2D). T. cimi sp. nov.
23. Lateral process of anterior coxal fold (alp) broader .24
- Lateral process of anterior coxal fold (alp) slender, regularly curved, sickle-shaped .25
24. Lateral process of anterior coxal fold (alp) terminating in a very short external spine and a very long
internal one; mesal process of anterior coxal fold (amp) as long as alp; first femoral spine (fe 1)
very short, pointed; second femoral spine (fe 2) very long, as long as tibial spine (ti); an additional
lamella at both sides of base of fe 2 . T. rich cirdh offmani Pimvichai et al., 2009b
- Lateral process of anterior coxal fold (alp) flattened, apically curved laterad as a short spine,
lateral margin of alp slightly folded; mesal process of anterior coxal fold (amp) shorter than
alp, slender, straight, directed distad, pointed (Fig. 10A); the first femoral spine (fe 1) very
13
European Journal of Taxonomy 199: 1-37 (2016)
short, directed upward, situated above the second femoral spine ife 2); fe 2 very long, slender,
curved downward (Fig. 10C) . T. ursus sp. nov.
25. Mesal margin of lateral process of anterior coxal fold (alp) simple, without a caudad spine or crest;
mesal process of anterior coxal fold (amp) much shorter than alp , curved, pointed.
. T. enghoffi (Demange, 1989)
- Mesal margin of lateral process of anterior coxal fold (alp) caudad with a small spine or crest.26
26. Mesal margin of lateral process of anterior coxal fold (alp) with a small caudad crest; mesal
process of anterior coxal fold (amp) slightly shorter than alp , slightly sigmoid, pointed.
. T. bifurcus (Demange, 1986)
- Mesal margin of lateral process of anterior coxal fold (alp) with a short curved caudad spine; mesal
process of anterior coxal fold (amp) as long as alp , straight ...I demangei Pimvichai et al. , 2009b
Species descriptions
Class Diplopoda Blainville-Gervais, 1844
Order Spirostreptida Brandt, 1833
Family Harpagophoridae Attems, 1909
Subfamily Harpagophorinae Attems, 1909
Thyropygus Pocock, 1894
Thyropygus cimi sp. nov.
urn:lsid:zoobank.org:act:0252646C-AD4E-4B8B-8A7C-AEEA80DD4A72
Fig. 2A-E
Diagnosis
A species of the opinatus subgroup. Lateral process of anterior coxal fold (alp) regularly curved, basally
broad, gradually tapering towards end and ending in sharp point, with a lateral crest near tip. Similar
in this respect to T. bifurcus. Differs from this species by having the mesal process of anterior coxal
fold (amp) straight, directed distad, by having the mesal process of posterior coxal fold (pmp) slender,
directed anteriad, and by having only one femoral spine (fe).
Etymology
This species is named after the organization “Centre International de Myriapodologie - CIM” ( www.
mvriapodologv.org ) in recognition of its imm ense importance for inspiring and supporting research on
myriapods.
Material examined
Holotype
THAILAND: Nakhonsrithammarat Province, Lanska District, Namwang Srithammasokrach,
8°20'25" N, 99°49'42" E, 16 May 2010, leg. S. Panha, P. Pimvichai and members of the Anim al
Systematics Research Unit (CUMZ-D00086).
Paratypes
THAILAND: 2 $$, 1 §, same data as holotype (CUMZ-D00096); 1 S, same data as holotype (ZMUC).
Description
Adult males with 65-68 podous rings, no apodous rings. Length 13-16 cm, width 7.6-8.3 mm. Adult
female with 65 podous rings, no apodous rings. Length 13 cm, width 7.4 mm. Colour in life unknown;
14
PIMVICHAIP. et al ., New species of Thyropygus millipedes
preserved specimens with head, antennae, prozona, middorsal metazona and legs brown; metazona,
epiproct, paraprocts and hypoproct dark brown.
Gonopods (Fig. 2A-E). Anterior coxal fold ( ac ; Fig. 2A): lateral process (alp) regularly curved, basally
broad, gradually tapering towards end and ending in sharp point, with lateral crest near tip (Fig. 2C,
arrow); mesal process (amp) slightly shorter than alp , straight, directed distad. Posterior coxal fold
(pc; Fig. 2B) basally with moderately high lateral paracoxites (px), distally truncate, forming shelf for
accommodation of telopodite, mesal process (pmp) slender, directed anteriad. Telopodite (Fig. 2D-E)
leaving coxite over shelf of posterior coxal fold; femoral spine (fe) very long, slender, curving downward,
in situ resting between alp and amp; tibial spine (ti) long, slender, curving in horizontal plane, its tip
C
D
E
E
E
E
E
Fig. 2. Thyropygus cimi sp. nov., from Namwang Srithammasokrach, holotype (CUMZ-D00086),
S, gonopods. A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed.
C. Lateral view. D. Left telopodite, posterior-mesal view. E. Left telopodite, anterior-lateral view.
15
European Journal of Taxonomy 199: 1-37 (2016)
in situ resting close to base of fe\ apical part: lamellar lobe ( II) broadly expanded, bent down; palette
(pa) simple, gutter-like, with longitudinal rounded crest (cr) at middle; distally with about ten brownish
blepharochaetae (bp).
DNA barcode
The GenBank accession number of the barcode of the holotype is KU306519 (voucher code
CUMZ-D00086).
Distribution (Fig. 12)
Known only from the type locality.
Remarks
Coexisting with the smaller T. forceps sp. nov.
Thyropygus culter sp. nov.
urn:lsid:zoobank.org:act:0093C39D-FlFC-4CAD-9DA0-0492F45E7370
Fig. 3A-D
Diagnosis
A species of the opinatus subgroup. Lateral process of anterior coxal fold (alp) flattened, slightly
curved, its laterodistal margin coarsely dentate. Similar in this respect to T. cristagalli , T. implicatus
and T. undulatus sp. nov. Differs from these species by having the mesal process of posterior coxal fold
(pmp) very high, pointed-triangular, directed almost straightly distad, and by having the tibial spine (ti)
recurved.
Etymology
The name is a Latin noun in apposition, meaning “knife”, and refers to the knifelike second femoral
spine (fe 2).
Material examined
Holotype
THAILAND: S, Krabi Province, Khlong Thom District, Rorn waterfall, 7°56'06" N, 99°12T8" E,
15 Jan. 2009, leg. P. Pimvichai, C. Sutcharit and members of the Animal Systematics Research Unit
(CUMZ-D00091).
Paratypes
THAILAND: 3 2 $ §, 1 subadult §, same data as holotype (CUMZ-D00078); 1 1 $, same data
as holotype (ZMUC).
Description
Adult males with 58-62 podous rings, no apodous rings. Length 12-14 cm, width 7.1-8.0 mm. Adult
females with 59-62 podous rings, no apodous rings. Length 11-13 cm, width 6.9-7.9 mm. Colour in
life unknown; preserved specimens with head, antennae, prozona, middorsal metazona and legs brown;
metazona, epiproct, paraprocts and hypoproct reddish brown.
Gonopods (Fig. 3A-D). Anterior coxal fold (ac; Fig. 3A): lateral process (alp) flattened, slightly curved,
its laterodistal margin coarsely dentate, terminating in short, sharp, pointed spine; mesal process (amp)
much shorter than alp , directed distad, tip curving mesad, pointed. Posterior coxal fold (pc\ Fig. 3B)
basally with lateral paracoxites (px) quite low, distally with two processes: mesal process (pmp ) very
16
PIMVICHAIP. et al ., New species of Thyropygus millipedes
high, pointed-triangular, directed almost straightly distad; lateral process {pip) much shorter, digitiform,
directed distad and slightly laterad. Telopodite (Fig. 3C-D) leaving coxite over shelf of posterior coxal
fold; femoral spine (fe) duplicated,^ 1 projecting above top of telopodite curvature, curving backward,
ending in slender barb,ye 2 basally broad, situated under fe 1 , curving backward, ending in slender barb;
Fig. 3. Thyropygus culter sp. nov., from Rom waterfall, holotype (CUMZ-D00091), S, gonopods.
A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed. C. Left telopodite,
posterior-mesal view. D. Left telopodite, anterior-lateral view.
17
European Journal of Taxonomy 199: 1-37 (2016)
tibial spine (ti) long, slender and recurved (not curving in horizontal plane); apical part: spatulate lobe
(si) slender, rounded; with slender, long spine (sis) at base of apical part, opposite origin of tibial spine;
palette (pa) simple, distally with about twelve brownish blepharochaetae (bp).
DNA barcode
The GenBank accession number of the barcode of one of the paratypes is KC519535 (voucher code
CUMZ-D00078).
Distribution (Fig. 12)
Known only from the type locality.
Thyropygus forceps sp. nov.
um:lsid:zoobank.org:act:llBAAEC5-404D-4E15-982C-2F3E51DEA56B
Figs 4A-E, 11B
Diagnosis
A species of the opinatus subgroup. Lateral process of anterior coxal fold (alp) long, slender, regularly
curved, tip close to tip of opposite alp , the two together forming a circle. Similar in this respect to
T. erectus and T. floweri. Differs from the former by having a telopodite lobe (lo). Particularly similar to
T. floweri , differing from it by having the mesal process of posterior coxal fold (pmp) slender, directed
distolaterad, laterally with a digitiform process (pip), and by not having pmp strongly developed along
the anterior-posterior axis.
Etymology
The name is a Latin noun in apposition, referring to the forceps-like gonopod coxae.
Material examined
Holotype
THAILAND: $, Nakhonsrithammarat Province, Lanslca District, Namwang Srithammasokrach,
8°20'25" N, 99°49'42" E, 16 May 2010, leg. S. Panha, P. Pimvichai and members of the Anim al
Systematics Research Unit (CUMZ-D00092).
Paratypes
THAILAND: 2 2 subadult $$, same data as holotype (CUMZ-D00073); 5 $$, Nakhonsrithammarat
Province, Ronpiboon District, Tham Pha Deang temple, 8°14'42" N, 99°52'03" E, 14 May 2012, leg. C.
Sutcharit and members of the Animal Systematics Research Unit (CUMZ-D00093); 2 $$, same data
as preceding (ZMUC).
Description
Adult males with 54-62 podous rings, no apodous rings. Length 9-12 cm, width 4.6-5.8 mm. Overall
colour of living animal (Fig. 11B) brown. Legs, antennae, middorsal metazona, epiproct, paraprocts
and hypoproct brownish orange; preserved specimens with head, antennae, prozona, legs, middorsal
metazona, epiproct, paraprocts and hypoproct brown; metazona dark brown.
Gonopods (Fig. 4A-E). Anterior coxal fold (ac; Fig. 4A): distinctly crenulated along convex part of
lateral margin (smooth in specimens from Tham Pha Deang temple), lateral process (alp) long, slender,
regularly curved, tip close to tip of opposite alp, the two together forming a circle; mesal process (amp)
slightly shorter than alp, directed obliquely disto-mesad (in specimens from Tham Pha Deang temple
lateral process (alp) long, slender, crossing over with opposite tip, the two together forming a circle;
18
PIMVICHAIP. et al ., New species of Thyropygus millipedes
Fig. 4B); mesal process (amp) slightly shorter than alp, directed distad, slightly sigmoid. Posterior coxal
fold (pc ; Fig. 4C) basally with moderately high lateral paracoxites (px), distally with two processes:
mesal process (pmp) slender, directed distolaterad; lateral process (pip) digitiform. Telopodite
(Fig. 4D-E) leaving coxite over shelf of posterior coxal fold; femoral spine (fe) massive, curving in
horizontal plane and curving against tv, telopodite distally to fe with large, round lobe (lo) projecting
distolaterally; tibial spine (ti) very long, slender, curving in horizontal plane, its tip resting against base
of fe ; apical part: spatulate lobe (si) with sharp dark brown spine at tip; palette (pa) simple, distally with
about nine brownish blepharochaetae (bp).
Fig. 4. Thyropygusforceps sp. nov., gonopods. - A, C-E. Holotype (CUMZ-D00092), $, fromNamwang
Srithammasokrach. A. Anterior view, left telopodite removed. C. Posterior view, left telopodite removed.
D. Left telopodite, posterior-mesal view. E. Left telopodite, anterior-lateral view. - B. Specimen from
Tham Pha Deang temple (CUMZ-D00093), $. Anterior view, left telopodite removed.
19
European Journal of Taxonomy 199: 1-37 (2016)
DNA barcode
The GenBank accession number of the barcode of one of the paratypes is KC519531 (voucher code
CUMZ-D00073).
Distribution (Fig. 12)
Known only from Namwang Srit.ha. mm asokrach and Tham Pha Deang temple in Nakhonsritha. mm a.rat
Province.
Remarks
Coexisting with the larger T. cimi sp. nov. at Namwang Srithammasokrach.
Thyropygus mesocristatus sp. nov.
um:lsid:zoobank.org:act:5D8F7830-2E8E-46E3-B2BE-830FC31F290D
Fig. 5A-E
Diagnosis
A species of the opinatus subgroup. Lateral process of anterior coxal fold (alp) slender, curving mesad;
mesal process of anterior coxal fold (amp) as long as alp , slender, straight, directed distad. Similar
in these respects to T. demangei. Differs from this species by having mesal margin of amp forming a
strong longitudinal crest {Ic) in posterior view, by having only one femoral spine (fe) and by having the
spatulate lobe ( si) broad and rounded.
Etymology
The name is a Latin adjective, referring to the longitudinal crest on the mesal process of the anterior
coxal fold.
Material examined
Holotype
THAILAND: f, Songkhla Province, Rattaphum District, Srikasorn, 7°00'18" N, 100°08'24" E, 12
Jan. 2009, leg. P. Pimvichai, C. Sutcharit and members of the Animal Systematics Research Unit
(CUMZ-D00094).
Paratype
THAILAND: S, same data as holotype (CUMZ-D00077).
Description
Adult males with 61-66 podous rings, no apodous rings. Length 16-17 cm, width 9.2-9.5 mm. Colour
in life unknown; preserved specimens with head, antennae, prozona and legs brown; metazona, epiproct,
paraprocts and hypoproct reddish brown.
Gonopods (Fig. 5A-E). Anterior coxal fold ( ac ; Fig. 5A): lateral process (alp) slender, curving mesad,
with lateral serrate crest near tip (Fig. 5C, arrow); mesal process (amp) as long as alp , slender, straight,
directed distad, tip sharp, pointed, in posterior view mesal margin of amp forming strong longitudinal
crest (lc). Posterior coxal fold ( pc ; Fig. 5B) basally with moderately high lateral paracoxites (px),
distally with two processes: mesal process (pmp) slender, directed anteriad; lateral process (pip) short,
erect, digitiform; forming shelf for accommodation of telopodite. Telopodite (Fig. 5D-E) leaving coxite
between pmp and plp\ femoral spine (fe) long, curved downward, with expanded lamella at base; tibial
spine (ti) very long, curving in horizontal plane, tip close to basal part of fe\ small spine (ss) at base of
apical part, opposite origin of tibial spine; spatulate lobe (si) broad, rounded; a small accessory lobe (al)
20
PIMVICHAIP. et al ., New species of Thyropygus millipedes
at base of si: palette (pa ) simple, gutter-like, with longitudinal crest (cr) at middle, distally with about
twelve brownish blepharochaetae (bp).
DNA barcode
The GenBank accession number of the barcode of the paratype is KC519534 (voucher code
CUMZ-D00077).
Distribution (Fig. 12)
Known only from the type locality.
Fig. 5. Thyropygus mesocristatus sp. nov., from Srikasorn, holotype (CUMZ-D00094), S, gonopods.
A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed. C. Lateral view.
D. Left telopodite, posterior-mesal view. E. Left telopodite, anterior-lateral view.
21
European Journal of Taxonomy 199: 1-37 (2016)
Thyropygus navychula sp. nov.
um:lsid:zoobank.org:act:01AC06C9-EEF0-4211-9837-28FE2C12C68E
Figs 6A-D, 11A
Diagnosis
A species of the opinatus subgroup. Mesal margin of anterior coxal fold (alp) with fine serrations.
Similar in this respect to T. opinatus. Differs from this species by having the lateral process (alp)
Fig. 6. Thyropygus navychula sp. nov., from Surin Islands, holotype (CUMZ-D00095), <$, gonopods.
A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed. C. Left telopodite,
posterior-mesal view. D. Left telopodite, anterior-lateral view.
22
PIMVICHAIP. et al. , New species of Thyropygus millipedes
slender, regularly curved and by having the mesal process (amp) broadly expanded and forming a strong
longitudinal crest (Ic) in posterior view.
Etymology
The species is named after the Royal Thai Navy, in recognition of their kind assistance which enabled
us to pursue the necessary fieldwork at the type locality, and also after Chulalongkorn University where
pink is the symbolic colour of the university, indirectly referring to the pink legs of the species; the
species name is treated as a noun in apposition.
Material examined
Holotype
THAILAND: $, Phang-Nga Province, Khuraburi District, Surin Islands, 9°24'20" N, 97°51'38" E,
7 Apr. 2012, leg. S. Panha, P. Pimvichai and members of the Animal Systematics Research Unit
(CUMZ-D00095).
Paratypes
THAILAND: 5 $$, 1 §, same data as holotype (CUMZ-D00089); 2 same data as holotype
(ZMUC).
Description
Adult males with 58-62 podous rings, no apodous rings. Tength 11-12 cm, width 6.2-7.2 mm. Adult
female with 60 podous rings, no apodous rings. Tength 14 cm, width 8.0 mm. Overall colour of living
animal (Fig. 11 A) dark brown, shiny. Tegs and antennae pink; preserved specimen with head, antennae,
prozona, legs, epiproct, paraprocts and hypoproct brown; metazona dark brown.
Gonopods (Fig. 6A-D). Anterior coxal fold ( ac ; Fig. 6A): lateral process (alp) slender, regularly curved,
mesal margin with fine serrations; mesal process (amp) almost as long as alp , broadly expanded, apically
sharp, straight distad, in posterior view mesal margin of amp forming strong longitudinal crest (Ic).
Posterior coxal fold ( pc ; Fig. 6B) basally with moderately high lateral paracoxites (px), distally with
two processes: mesal process (pmp) shorter than pip , curving behind pip: lateral process (pip) flattened,
directed distad. Telopodite (Fig. 6C-D) leaving coxite between pmp and pip: femoral spine (fe) very
long, curving almost exclusively in horizontal plane; tibial spine (ti) very long, curving in horizontal
plane close to basal part of fe: spatulate lobe (si) terminating in sharp spine; palette (pa) simple, gutter¬
like, distally with about twelve brownish blepharochaetae (bp), laterally with few sharp spines.
DNA barcode
The GenBank accession number of the barcode of one of the paratype is KU306522 (voucher code
CUMZ-D00089).
Distribution (Fig. 12)
Known only from the type locality.
Thyropygus planispina sp. nov.
um:lsid:zoobank.org:act:2F5A25A4-44CC-4B93-AC17-9A9B6E6E9ElF
Fig. 7A-E
Diagnosis
A species of the opinatus subgroup. Differs from all other species in the subgroup by having the lateral
process of the anterior coxal fold (alp) regularly curved, terminating in a sharp spine pointing slightly
distad, and by having the tibial spine (ti) short and flattened.
23
European Journal of Taxonomy 199: 1-37 (2016)
Etymology
The name is a Latin noun in apposition and refers to the flattened tibial spine (ti).
Material examined
Holotype
THAILAND: S, Krabi Province, Muang District, Tham Sua temple, 8°07'30" N, 98°55'30" E, 15
Jan. 2012, leg. P. Pimvichai, C. Sutcharit and members of the Animal Systematics Research Unit
(CUMZ-D00088).
Fig. 7. Thyropygus planispina sp. nov., from Tham Sua temple, holotype (CUMZ-D00088), S, gonopods.
A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed. C. Lateral view.
D. Left telopodite, posterior-mesal view. E. Left telopodite, anterior-lateral view.
f e 2
24
PIMVICHAIP. et al. , New species of Thyropygus millipedes
Paratypes
THAILAND: 3 same data as holotype (CUMZ-D00097).
Description
Adult male with 64 podous rings, no apodous rings. Length 13 cm, width 7.6 mm. Adult females with
60-63 podous rings, no apodous rings. Length 13-14 cm, width 7.8-8.6 mm. Colour in life unknown;
preserved specimens with head, antennae, legs, metazona, epiproct, paraprocts and hypoproct brown;
prozona yellowish brown.
Gonopods (Fig. 7A-E). Anterior coxal fold (ac; Fig. 7A): lateral process (alp) regularly curved,
terminating in sharp spine pointing slightly distad, with lateral short, sharp crest at middle of alp
(Fig. 7C, arrow); mesal process (amp) slightly shorter than alp , flattened, straight, directed distad.
Posterior coxal fold (pc ; Fig. 7B) basally with moderately high lateral paracoxites (px), distally truncate,
forming shelf for accommodation of telopodite, mesal process (pmp) triangular, directed distad, lateral
process (pip) erect, digitiform. Telopodite (Fig. 7D-E) leaving coxite over shelf of posterior coxal fold;
femoral spine double,^ 1 very long, curved downward, situated above fe 2 , in situ resting at middle of
alp, fe 2 long, very slender, curved downward; tibial spine (ti) flattened, short, curving mesad; apical
part: spatulate lobe (si) small, rounded; with slender, long spine (sis) at base of apical part; palette (pa)
simple, gutter-like, distally with about nine brownish blepharochaetae (bp).
DNA barcode
The GenBank accession number of the barcode of the holotype is KU306521 (voucher code
CUMZ-D00088).
Distribution (Fig. 12)
Known only from the type locality.
Thyropygus sutchariti sp. nov.
urn:lsid:zoobank.om:act:6804DF2C-7787-46FE-9185-2E5C40AF5ED3
Fig. 8A-D
Diagnosis
A species of the opinatus subgroup. Mesal process of anterior coxal fold (amp) directed slightly obliquely
disto-mesad, tip overlapping tip of opposite amp. Similar in this respect to T. chelatus. Differs from this
species by having amp as long as alp and by having the lateral process of the posterior coxal fold (pip)
as a massive, broad lobe.
Etymology
The species is named in honour of Chirasak Sutcharit in recognition of his devotion to collecting
millipedes.
Material examined
Holotype
THAILAND: Phetchaburi Province, Kaeng Krachan District, Kaeng Krachan, 12°54'07" N,
99°35'56" E, 14 May 2012, leg. C. Sutcharit and members of the Animal Systematics Research Unit
(CUMZ-D00090).
Paratypes
THAILAND: 2 SS, 2 55, 1 subadult 5, same data as holotype (CUMZ-D00098); 1 S, same data as
holotype (ZMUC).
25
European Journal of Taxonomy 199: 1-37 (2016)
Description
Adult males with 63-65 podous rings, no apodous rings. Length 10-11 cm, width 5.4-5.7 mm. Adult
females with 61-63 podous rings, no apodous rings. Length 12-13 cm, width 6.7-6.9 mm. Colour in life
unknown; preserved specimens with head, antennae, prozona, legs, epiproct, paraprocts and hypoproct
brown; metazona reddish brown.
E
E
C
E
E
D
Fig. 8. Thyropygns sutchariti sp. nov., from Kaeng Krachan, holotype (CUMZ-D00090), 3, gonopods.
A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed. C. Left telopodite,
posterior-mesal view. D. Left telopodite, anterior-lateral view.
26
PIMVICHAIP. et al., New species of Thyropygus millipedes
Gonopods (Fig. 8A-D). Anterior coxal fold ( ac ; Fig. 8A): lateral process {alp) slender, regularly curved;
mesal process {amp) as long as alp, directed slightly obliquely disto-mesad, tip overlapping tip of
opposite amp. Posterior coxal fold {pc. Fig. 8B) basally with moderately high lateral paracoxites {px),
distally with two processes: mesal process {pmp) a rounded lobe; lateral process {pip) a massive, broad
lobe, projecting laterad. Telopodite (Fig. 8C-D) leaving coxite between pmp and pip ; femoral spine (fe)
long, curved downward; telopodite distally to fe with large, round lobe {lo) projecting distolaterally;
tibial spine {ti) very long, curving in horizontal plane, tip close to basal part of lo; spatulate lobe {si)
broad, rounded; palette {pa) simple, gutter-like, distally with about ten brownish blepharochaetae {bp).
DNA barcode
The GenBank accession number of the barcode of the holotype is KU306524 (voucher code
CUMZ-D00090).
Distribution (Fig. 12)
Known only from the type locality.
Thyropygus undulatus sp. nov.
um:lsid:zoobank.org:act:972296F7-CF7B-4AD4-A2AA-3DCE22596B3E
Fig. 9A-E
Diagnosis
A species of the opinatus subgroup. Lateral process {alp) flattened, slightly curved, its laterodistal margin
coarsely dentate. Similar in this respect to T. cristagaUi, T. implicatus and T. culter sp. nov. Differs from
these species by having the mesal process of the posterior coxal fold {pmp) with two rounded distal
lobes, visible in anterior view between alp and amp, and by having the tibial spine (ti) not ending in a
sharp point.
Etymology
The name is a Latin adjective, referring to the undulate/coarsely dentate laterodistal margin of the lateral
process of the anterior coxal fold.
Material examined
Holotype
THAILAND: $, Krabi Province, Muang District, Khao Phanom Bencha, 8°16'30" N, 98°55'30" E,
16 Jan. 2009, leg. P. Pimvichai, C. Sutcharit and members of the Animal Systematics Research Unit
(CUMZ-D00087).
Paratypes
THAILAND: 1 5, same data as holotype (CUMZ-D00099).
Description
Adult male with 63 podous rings, no apodous rings. Length 15 cm, width 8.7 mm. Adult female with
64 podous rings, no apodous rings. Length 14 cm, width 8.5 mm. Colour in life unknown; preserved
specimens with head, legs, middorsal metazona, epiproct, paraprocts and hypoproct brown; antennae
and metazona dark brown; prozona whitish-brown.
Gonopods (Fig. 9A-E). Anterior coxal fold {ac; Fig. 9A): lateral process {alp) flattened, curving mesad,
its laterodistal margin coarsely dentate, terminating in a short spine, tip curving against tip of opposite
side, lateral margins with serrate crest (Fig. 9C, arrow); mesal process {amp) much shorter than alp.
27
European Journal of Taxonomy 199: 1-37 (2016)
slender, curving mesad. Posterior coxal fold ( pc ; Fig. 9B) basally with moderately high lateral paracoxites
(px): mesal process ( pmp ) distally with two rounded lobes, visible in anterior view between alp and amp:
lateral process {pip ) erect, digitiform. Telopodite (Fig. 9D-E) leaving coxite over shelf of posterior
coxal fold; femoral spine (fe) duplicated,/^ 1 relatively long, projecting above top of telopodite, curving
backward, ending in slender barb, 7^ 2 basally broad, situated under fe 1 , curving backward, ending in
slender barb; tibial spine (ti) long, curving in horizontal plane, not ending in sharp spine; apical part:
spatulate lobe (si) rounded, with slender long spine (sis) at base of apical part, opposite origin of tibial
spine; palette (pa) simple, distally with about eleven brownish blepharochaetae (bp).
DNA barcode
The GenBank accession number of the barcode of the holotype is KU306520 (voucher code
CUMZ-D00087).
Fig. 9. Thyropygus undulatus sp. nov., from Khao Phanom Bencha, holotype (CUMZ-D00087), /,
gonopods. A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed.
C. Lateral view. D. Left telopodite, posterior-mesal view. E. Left telopodite, anterior-lateral view.
28
Distribution (Fig. 12)
Known only from the type locality.
PIMVICHAIP. et al ., New species of Thyropygus millipedes
Thy ropygus ursus sp. nov.
um:lsid:zoobank.org:act:D3E091F2-C075-458B-A9B5-DA35B673309B
Fig. 10A-D
Diagnosis
A species of the opinatus subgroup. Differs from all other species in the subgroup by having the lateral
process of the anterior coxal fold (alp) flattened, broad, apically curved caudad and ending in a short
spine, as well as by the first femoral spine (fe 1) being very short and erect.
fe 2
E
E
Fig. 10. Thyropygus ursus sp. nov., from Lanta Islands, holotype (NMHW-Inv.7855), gonopods.
A. Anterior view, left telopodite removed. B. Posterior view, left telopodite removed. C. Left telopodite,
posterior-mesal view. D. Left telopodite, anterior-lateral view.
29
European Journal of Taxonomy 199: 1-37 (2016)
Etymology
The name is a Latin noun in apposition, meaning “bear”, and refers to the (somewhat) bearhead-like
profile of the lateral process of the anterior coxal fold (alp).
Material examined
Holotype
THAILAND: $, Krabi Province, Koh Lanta District, Lanta Islands, 7°39'20" N, 99°02'31" E, 2 Jan.
2009, leg. B. Daubl and E. Haring (NMHW-Inv.7855).
Paratypes
THAILAND: 1 §, same data as holotype (NMHW).
Description
Adult male with 62 podous rings, no apodous rings. Length 11 cm, width 6.4 mm. Adult female with
61 podous rings, no apodous rings. Length 12 cm, width 7.4 mm. Colour in life unknown; preserved
specimens with head, antennae, legs, metazona, epiproct, paraprocts and hypoproct brown; prozona
yellowish brown.
Gonopods (Pig. 10A-D). Anterior coxal fold (ac; Pig. 10A): lateral process (alp) flattened, broad, apically
curved caudad, tip with short spine, lateral margin slightly folded; mesal process (amp) shorter than alp,
slender, straight, directed distad, pointed. Posterior coxal fold (pc; Pig. 10B) basally with moderately
high paracoxites (px), distally truncate, forming shelf to accommodate telopodite, mesal process (pmp )
very small, directed distolaterad. Telopodite (Pig. 10C-D) leaving coxite over shelf of posterior coxal
fold; femoral spine double, fe 1 very short, erect, situated above fe 2, fe 2 very long, slender, curved
downward, in situ resting behind alp; tibial spine (ti) long, slender, curving in horizontal plane, its tip
close to base of fe; apical part: lamellar lobe (II) broad, bent down; palette (pa) simple, gutter-like, with
small crest (cr) near tip; distally with about twelve brownish blepharochaetae (bp).
DNA barcode
The GenBank accession number of the barcode of the holotype is KU306523 (voucher code NMHW-
Inv.7855).
Distribution (Fig. 12)
Known only from the type locality.
Fig. 11. A. Thyropygus navychula sp. nov., specimen from Surin Islands, living $ (paratype,
CUMZ-D00089-1). B. Thyropygus forceps sp. nov., specimen from Namwang Srithammasokrach,
living f (paratype, CUMZ-D00073-1).
30
PIMVICHAIP. et al. , New species of Thyropygus millipedes
Discussion
In our previous treatment of the T. opinatus subgroup (Pimvichai et al. 2009a) we anticipated that
DNA sequence data would help resolve relationships between Thyropygus species. A subsequent study
(Pimvichai et al. 2014), which included a mtDNA sequence analysis of 23 Thyropygus species, did
indeed shed light over these relationships. The present study expands the taxonomic coverage of this
latter analysis by adding 9 new species to the mtDNA dataset. The analysis clearly shows that the
opinatus and bifurcus subgroups cannot be maintained as separate groups and hence should be merged
into an extended opinatus subgroup. The monophyly of this (extended) opinatus subgroup is well-
supported (Fig. 1). However, the phylogenetic relationships within the opinatus subgroup are often not
well-resolved. Thus, still more species and nuclear genes need to be added to improve the phylogenetic
resolution.
For the delimitation of species in the opinatus subgroup we used both gonopodal characters and
divergences in COI sequences (Table 2). Interspecific distances among Bavarian Diplopoda ranged
rm
Distribution of the opinatus subgroup
T. bearti
T. bifurcus
T. bispinispatula
T. bispin us
T. brachyacanthus
T. casjeekeli
T. chelatus
T. cimi sp. nov.
T. cristagalli
7, culter sp. nov
T. demangei
7. enghoffi
T. erectus
T. floweri
T. forceps sp. nov.
7. implicatus
T. inflexus
T. loxia
T. mesocristatus sp. nov.
7. navychuia sp. nov.
7. opinatus
T. planispina sp. nov.
7. quadricuspis
7. richardhoffmani
T. sutchariti sp. nov.
7. undulatus sp. nov.
7, ursus sp. nov.
Fig. 12. Known distribution of the species of the T. opinatus subgroup.
31
European Journal of Taxonomy 199: 1-37 (2016)
between 0% (in subspecies) and 33.18 % (among different orders). The mean value of the interspecific
distance for Diplopoda was 14.17% (Spelda et al. 2011). For 21 species of the opinatus subgroup the
divergence was between 2 and 17%. The lowest interspecific divergence was 2%, between T. cristagalli
and T. planispina sp. nov., between T. planispina sp. nov. and T. undulatus sp. nov., and between
T. quadricuspis and T. ursus sp. nov. In these cases, speciation may have been very recent but we still need
more DNA data and more samples to be able to infer the taxonomic status and relationships among these
species properly. However, all these species show unique gonopodal characters and occur in separate
localities and hence, for the time being, we treat these “morphs” as separate species. Nevertheless,
intraspecific divergence of T. induratus was also 2% in the samples of the present study but despite
this sequence divergence the overall gonopod configuration remained the same in the specimens here
assigned to T. induratus.
All species of the opinatus subgroup share a conspicuous synapomorphy, viz. the additional projection
on the anterior coxal fold. This character separates the opinatus subgroup from other subgroups of the
T. allevatus group.
The main gonopodal variations within the opinatus subgroup are:
- the number of femoral spines (fe)\ one or two
- presence/absence of a spatulate lobe (si)
- presence/absence of a longitudinal rounded crest (cr) near the tip of the palette
- presence/absence of a small spine (ss) or a slender long spine (sis) at the posterior base of the apical
part of the telopodite
- presence/absence of a lateral (pip) and a mesal (pmp) process of the posterior coxal fold
The vast majority of the species we have studied and of the previous records of the T. opinatus subgroup
are from the southern part of Thailand (Fig. 12). The remaining material comes from the North and West
of Thailand and from some adjacent areas in Myanmar and Malaysia. Most probably further fieldwork
in all these areas will lead to the discovery of further new species of Thyropygus .
Acknowledgements
This research was funded by the Thailand Research Fund (TRF-MRG5680056) to PR PP was also
funded by an EC Marie Curie Postdoctoral Fellowship to non EU-researchers issued by the Belgian
Science Policy Office (BELSPO). Additional funding came from the Thailand Research Fund (TRF) as a
TRF Senior Research Scholar (2015-2018; RTA 5880002) to SP. We express our sincere gratitude to the
Plant Genetic Conservation Project initiated by her Royal Hig hn ess Princess Maha Chakri Sirindhorn,
and the Navy Special Warfare Unit of the Royal Thai Navy for their kind assistance and encouragement,
which enabled us to pursue the necessary fieldwork at Surin Islands and Lanta Islands, Phang-Nga
Province. We also thank members of the Animal Systematics Research Unit, Chulalongkorn University
for assistance in collecting specimens, especially to Dr. Chirasalc Sutcharit, to Dr. Verena Stagl (NHMW)
who provided specimens from Lanta Islands, and to Thita Krutchuen for the excellent drawings.
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Topic editor: Rudy Jocque
Desk editor: Danny Eibye-Jacobsen
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EJT consortium: Museum national d’Histoire naturelle, Paris, France; Botanic Garden Meise, Belgium;
Royal Museum for Central Africa, Tervuren, Belgium; Natural History Museum, London, United
Kingdom; Royal Belgian Institute of Natural Sciences, Brussels, Belgium; Natural History Museum of
Denmark, Copenhagen, De nm ark
34
PIMVICHAIP. et al ., New species of Thyropygus millipedes
Appendix I
Estimates of evolutionary divergence between 16S rRNA sequences expressed as p-distance.
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T. allevatus Bb
T. allevatus Bk
T. bearti Ks
T. bispinispatula Kb
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T. brachyacanthus Sk
T. cimi sp. nov. NWS1_2
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35
European Journal of Taxonomy 199: 1-37 (2016)
Appendix II
Phylogenetic relationships of Thyropygus species based on maximum likelihood analysis (ML) and
Bayesian Inference (BI) of partial COI gene (660 bp). Numbers at nodes indicate branch support based
on bootstrapping (ML) / posterior probability (BI). Scale bar = 0.07 substitutions/site. # indicates
branches which received < 50% ML bootstrap support and < 0.70 BI posterior probability, - indicates
non-supported branches by posterior probability. Abbreviations after species names refer to locality
names as shown in Table 1.
98 / 1.00
98 / 1.00
- Anurostreptus sculptus PI
■Anurostreptus barthelemyae Tib
100 / 1.00
57 / 0.96
Thyropygus allevatus Bk
Thyropygus allevatus Bb
i oo/i .oo j Thyropygus resimus Kc
■Thyropygus resimus Ksp
loo/i.oo i Thyropygus uncinatus Rcb
Thyropygus uncinatus Kdb
I —Thyropygus induratus Nru
'Thyropygus induratus Tt
Thyropygus foliaceus Kj
Thyropygus jarukchusri Np
Thyropygus inflexus Kk
Thyropygus bispinus Tm
m\
51 / 0.82
70 / 0.95
Thyropygus cutter sp. nov. Nr
Thyropygus mesocristatus sp. nov. Ss
Thyropygus bearti Ks
|- Thyropygus cimi sp. nov. Nws1_2
r]p Thyropygus quadricuspis Hy
Thyropygus ursus sp. nov. LTK
Thyropygus demangei Cm
Thyropygus richardhoffmani TCH
Thyropygus enghoffi Kl
■ 9 . 2 . /1 .-°l| 86/0 jP jhyropygus planispina sp. nov. Tsk2
Thyropygus undulatus sp. nov. Pb2
Thyropygus cristagalli Yn
Thyropygus brachyacanthus Sk
Thyropygus loxia Ty
Thyropygus sutchariti sp. nov. KCP
Thyropygus forceps sp. nov. Nws2
Thyropygus bispinispatula Kb
Thyropygus opinatus Arw
Thyropygus navychula sp. nov. KSR
0.07
36
PIMVICHAIP. et al ., New species of Thyropygus millipedes
Appendix III
Phylogenetic relationships of Thyropygus species based on maximum likelihood analysis (ML) and
Bayesian Inference (BI) of partial 16S rRNA gene (487 bp). Numbers at nodes indicate branch support
based on bootstrapping (ML) / posterior probability (BI). Scale bar = 0.06 substitutions/site. # indicates
branches which received < 50% ML bootstrap support and < 0.70 BI posterior probability, - indicates
non-supported branches by posterior probability. Abbreviations after species names refer to locality
names as shown in Table 1.
100/1.00
Anurostreptus barthelemyae Tib
• Anurostreptus sculptus PI
76 /o. 94 | Thyropygus bispinus Tm
76/0.99
70/0.96
100 / 1.00
#/#
#/#
71/0.80
■ Thyropygus inflexus Kk
Thyropygus sutchariti sp. nov. KCP
85/0.98
• Thyropygus loxia Ty
#/#
■ Thyropygus bispinispatula Kb
•Thyropygus richardhoffmani TCH
Thyropygus cimi sp. nov. Nws1_2
72/0.97
52/0.80
541-J— .
ft-7/
■■4 rTI
93/1.00
1 j- Thyropygus quadricuspis Hy
n* *-Thyropygus ursus sp. nov. LTK
L Thyropygus demangei Cm
—-Thyropygus cutter sp. nov. Nr
Thyropygus mesocristatus sp. nov. Ss
88 /o 95 ~ Thyropygus brachyacanthus Sk
j Thyropygus undulatus sp. nov. Pb2
J^Thyropygusplanispina sp. nov. Tsk2
UJI Thyropygus cristagalli Yn
I- Thyropygus enghoffi KI
Thyropygus bearti Ks
Thyropygus opinatus Arw
Thyropygus navychula sp. nov. KSR
Thyropygus forceps sp. nov. Nws2
96/1.00
Thyropygus foliaceus Kj
— Thyropygus jarukchusri Np
100 / 1.00
#/-
100/0.99
57/0.73
60/0.92
100 / 1.00
75/0.9
17
_j Thyropygus altevatus Bb
I Thyropygus altevatus Bk
Thyropygus uncinatus Kdb
Thyropygus uncinatus Rcb
r Thyropygus resimus Kc
Thyropygus resimus Ksp
Thyropygus induratus Nru
Thyropygus induratus It
0.06
37