Corresponding author: Varpu Vahtera (
Academic editor: M. Zapparoli
A new species of
Vahtera V, Stoev P, Akkari N (2020) Five million years in the darkness: A new troglomorphic species of
Located in the southeastern part of Romania not far from the Black Sea Coast, Movile Cave is the first known subterranean chemosynthesis-based ecosystem (
Despite its harsh living conditions, Movile Cave ecosystem is known to harbor a diverse and unique fauna. The cave hosts 51 invertebrate species, of which 34 species are endemic (
Five species of myriapods are hitherto discovered from the innermost parts of Movile viz.
Recently, we had the occasion to study freshly collected specimens of an undetermined species of the genus
All
Morphological terminology follows
Abbreviations:
Altogether 29 specimens from both inside and outside the Movile Cave were included in the phylogenetic analysis. Of these, 14 were sequenced in this study. Total DNA was extracted from the legs using NucleoSpinTissue kit (Macherey-Nagel) according to the standard protocol for human or animal and cultured cells. Samples were incubated overnight. One nuclear (28S rRNA) and two mitochondrial (cytochrome
Polymerase chain reaction (
Specimens used in the molecular phylogeny and their GenBank accession numbers (specimens sequenced in this study in bold). Institutional abbreviations:
Species | Lab code | Voucher ID number | Voucher | Country |
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16S | 28S |
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K3 |
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Romania |
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K4 |
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Romania |
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1a |
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Serbia |
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1b |
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Serbia |
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2 |
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Serbia |
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– |
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3 |
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Serbia |
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4 |
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Serbia |
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7 |
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Serbia |
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8 |
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Serbia |
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9 |
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Serbia |
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12 |
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Serbia |
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13 |
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Serbia |
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54a |
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Romania |
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57a |
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Romania |
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Germany |
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– | – | ||
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Germany |
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– | – | ||
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Germany |
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– | – | ||
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Germany |
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– | – | ||
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Germany |
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– | – | ||
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Germany |
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– | – | ||
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Germany |
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– | – | ||
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IZ-131458 |
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UK |
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Austria |
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– | – | |||
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Germany |
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– | – | |||
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Slovenia |
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– | – | |||
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Croatia |
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– | – | |||
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Austria |
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– | – | ||
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IZ-130582 |
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UK |
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IZ-130592 |
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UK |
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IZ-131446 |
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Spain |
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Most specimens included in the analysis had all three markers successfully sequenced. To obtain more geographic variation in the dataset, 15
Phylogenetic analysis was conducted using both parsimony and maximum likelihood as optimality criteria. Parsimony analysis was done with TNT v. 1.5 (
Uncorrected p-distances of aligned
A species morphologically similar to
Differences in morphological characters between
Morphological character |
|
|
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Body size (mm) | 25–50 | >46–52 |
Antennae length | Until posterior end of T3 | Until mid of T5 |
Antennal article 7 L/W (mm) | 0.5 × 0.25 | 1.0 × 0.5 |
Antennae: spines on basal articles | Present, numerous | Lacking or just a few |
Ultimate leg length | 7.65 mm | 13.25 mm |
Ultimate leg pretarsus (mm) | 0.25 | 1 |
Ultimate leg saw teeth on tibia and tarsus 1 | Tibia: 7–12 (usually 7–10); Tarsus: 3–5 | Tibia: 13–17; Tarsus: 5–6 |
Legs | Short, compact, pretarsus short | Strongly elongated, pretarsus long |
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Ovoid, small to medium sized (Fig. |
Strongly elongated, large (Fig. |
Forcipular trochanteroprefemur | With spines medially (4–6) | Without spines, just stout setae |
Coxopleural pore field | Approx. 2/3 of coxapleura; composed of less than 100 pores (86–90) | Approx. 4/5 of coxopleura; composed of more than 310 pores (317–320) |
Length (anterior margin of head plate to posterior margin of telson) approx. 52 mm (46 mm in an adult paratype) (Figs
The species epithet is a noun in apposition, meaning "king of the cave", referring to the species top position in the food chain of the Movile ecosystem.
The species is hitherto known only from the aphotic zone of the Cave Movile in the southern part of Romanian Dobrogea.
Parsimony analysis resulted in a single most-parsimonious (MP) tree of length 1586 steps (Fig.
The single most parsimonious tree of length 1586 steps with jackknife resampling values > 50% shown on the nodes. Branch lengths represent the number of optimized character-state changes.
Regarding the placement of
Likelihood tree with bootstrap values > 50% shown for each node.
When analyzed separately (only likelihood, tree not shown), the mitochondrial
Pairwise distances between the samples by each marker are shown in Tables
Estimates of evolutionary divergence between sequences.
All positions containing gaps and missing data were eliminated. There were a total of 556 positions in the final dataset. | ||||||||||||||||||||||||||||||
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1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 | 27 | 28 | 29 | ||
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0.248 | |||||||||||||||||||||||||||||
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0.248 | 0.000 | ||||||||||||||||||||||||||||
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0.248 | 0.000 | 0.000 | |||||||||||||||||||||||||||
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0.248 | 0.000 | 0.000 | 0.000 | ||||||||||||||||||||||||||
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0.248 | 0.000 | 0.000 | 0.000 | 0.000 | |||||||||||||||||||||||||
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0.248 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | ||||||||||||||||||||||||
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0.248 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | |||||||||||||||||||||||
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0.248 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | ||||||||||||||||||||||
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0.237 | 0.110 | 0.110 | 0.110 | 0.110 | 0.110 | 0.110 | 0.110 | 0.110 | |||||||||||||||||||||
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0.223 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.144 | ||||||||||||||||||||
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0.223 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.144 | 0.000 | |||||||||||||||||||
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0.225 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.149 | 0.005 | 0.005 | ||||||||||||||||||
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0.246 | 0.155 | 0.155 | 0.155 | 0.155 | 0.155 | 0.155 | 0.155 | 0.155 | 0.153 | 0.121 | 0.121 | 0.126 | |||||||||||||||||
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0.225 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.142 | 0.095 | 0.095 | 0.101 | 0.085 | ||||||||||||||||
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0.212 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.138 | 0.135 | 0.074 | 0.074 | 0.079 | 0.112 | 0.103 | |||||||||||||||
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0.225 | 0.146 | 0.146 | 0.146 | 0.146 | 0.146 | 0.146 | 0.146 | 0.146 | 0.142 | 0.092 | 0.092 | 0.097 | 0.122 | 0.117 | 0.040 | ||||||||||||||
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0.243 | 0.129 | 0.129 | 0.129 | 0.129 | 0.129 | 0.129 | 0.129 | 0.129 | 0.138 | 0.103 | 0.103 | 0.108 | 0.104 | 0.097 | 0.088 | 0.099 | |||||||||||||
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0.239 | 0.133 | 0.133 | 0.133 | 0.133 | 0.133 | 0.133 | 0.133 | 0.133 | 0.140 | 0.097 | 0.097 | 0.103 | 0.110 | 0.094 | 0.086 | 0.094 | 0.023 | ||||||||||||
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0.239 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.138 | 0.095 | 0.095 | 0.101 | 0.108 | 0.092 | 0.085 | 0.092 | 0.022 | 0.002 | |||||||||||
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0.239 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.138 | 0.095 | 0.095 | 0.101 | 0.108 | 0.092 | 0.085 | 0.092 | 0.022 | 0.002 | 0.000 | ||||||||||
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0.239 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.131 | 0.138 | 0.104 | 0.104 | 0.110 | 0.112 | 0.092 | 0.099 | 0.106 | 0.032 | 0.027 | 0.025 | 0.025 | |||||||||
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0.241 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.137 | 0.135 | 0.094 | 0.094 | 0.099 | 0.106 | 0.090 | 0.090 | 0.101 | 0.040 | 0.029 | 0.027 | 0.027 | 0.043 | ||||||||
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0.243 | 0.203 | 0.203 | 0.203 | 0.203 | 0.203 | 0.203 | 0.203 | 0.203 | 0.182 | 0.174 | 0.174 | 0.178 | 0.173 | 0.189 | 0.180 | 0.187 | 0.198 | 0.200 | 0.198 | 0.198 | 0.191 | 0.191 | |||||||
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0.228 | 0.178 | 0.178 | 0.178 | 0.178 | 0.178 | 0.178 | 0.178 | 0.178 | 0.167 | 0.182 | 0.182 | 0.185 | 0.174 | 0.167 | 0.176 | 0.183 | 0.180 | 0.173 | 0.173 | 0.173 | 0.185 | 0.182 | 0.169 | ||||||
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0.230 | 0.182 | 0.182 | 0.182 | 0.182 | 0.182 | 0.182 | 0.182 | 0.182 | 0.180 | 0.200 | 0.200 | 0.203 | 0.201 | 0.191 | 0.203 | 0.209 | 0.198 | 0.196 | 0.194 | 0.194 | 0.194 | 0.192 | 0.156 | 0.156 | |||||
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0.221 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.173 | 0.176 | 0.176 | 0.182 | 0.191 | 0.182 | 0.171 | 0.180 | 0.171 | 0.169 | 0.167 | 0.167 | 0.167 | 0.171 | 0.196 | 0.192 | 0.185 | ||||
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0.239 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.185 | 0.173 | 0.173 | 0.176 | 0.192 | 0.178 | 0.174 | 0.180 | 0.194 | 0.194 | 0.192 | 0.192 | 0.196 | 0.196 | 0.169 | 0.192 | 0.192 | 0.137 | |||
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0.255 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.192 | 0.185 | 0.189 | 0.189 | 0.192 | 0.201 | 0.203 | 0.189 | 0.203 | 0.201 | 0.205 | 0.207 | 0.207 | 0.201 | 0.196 | 0.173 | 0.207 | 0.180 | 0.187 | 0.165 | ||
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0.223 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.201 | 0.187 | 0.192 | 0.192 | 0.196 | 0.210 | 0.194 | 0.178 | 0.192 | 0.198 | 0.194 | 0.194 | 0.194 | 0.205 | 0.198 | 0.210 | 0.185 | 0.187 | 0.196 | 0.203 | 0.216 |
Estimates of evolutionary divergence between sequences. 16S: The number of base differences per site from between sequences are shown. The analysis involved 17 nucleotide sequences. All positions containing gaps and missing data were eliminated.
There were a total of 392 positions in the final dataset. | |||||||||||||||||
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1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | ||
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0.390 | ||||||||||||||||
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0.372 | 0.092 | |||||||||||||||
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0.372 | 0.092 | 0.000 | ||||||||||||||
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0.372 | 0.094 | 0.003 | 0.003 | |||||||||||||
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0.372 | 0.082 | 0.036 | 0.036 | 0.038 | ||||||||||||
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0.367 | 0.092 | 0.041 | 0.041 | 0.041 | 0.018 | |||||||||||
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0.372 | 0.089 | 0.043 | 0.043 | 0.043 | 0.020 | 0.008 | ||||||||||
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0.372 | 0.087 | 0.041 | 0.041 | 0.041 | 0.010 | 0.008 | 0.010 | |||||||||
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0.365 | 0.084 | 0.033 | 0.033 | 0.036 | 0.018 | 0.031 | 0.033 | 0.023 | ||||||||
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0.372 | 0.092 | 0.059 | 0.059 | 0.059 | 0.048 | 0.054 | 0.051 | 0.048 | 0.051 | |||||||
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0.372 | 0.099 | 0.066 | 0.066 | 0.066 | 0.054 | 0.059 | 0.056 | 0.054 | 0.059 | 0.013 | ||||||
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0.365 | 0.099 | 0.082 | 0.082 | 0.082 | 0.066 | 0.074 | 0.071 | 0.069 | 0.071 | 0.066 | 0.069 | |||||
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0.383 | 0.112 | 0.087 | 0.087 | 0.084 | 0.071 | 0.077 | 0.074 | 0.069 | 0.082 | 0.079 | 0.082 | 0.066 | ||||
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0.385 | 0.084 | 0.094 | 0.094 | 0.097 | 0.077 | 0.087 | 0.084 | 0.082 | 0.079 | 0.097 | 0.107 | 0.107 | 0.125 | |||
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0.355 | 0.217 | 0.209 | 0.209 | 0.212 | 0.214 | 0.227 | 0.224 | 0.222 | 0.219 | 0.224 | 0.227 | 0.232 | 0.235 | 0.232 | ||
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0.360 | 0.230 | 0.217 | 0.217 | 0.219 | 0.222 | 0.235 | 0.230 | 0.232 | 0.224 | 0.235 | 0.245 | 0.230 | 0.219 | 0.230 | 0.260 |
Estimates of evolutionary divergence between sequences. 28S: The number of base differences per site from between sequences are shown. The analysis involved 18 nucleotide sequences.
All positions containing gaps and missing data were eliminated. There were a total of 316 positions in the final dataset. | ||||||||||||||||||
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1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | ||
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0.187 | |||||||||||||||||
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0.187 | 0.000 | ||||||||||||||||
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0.190 | 0.003 | 0.003 | |||||||||||||||
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0.190 | 0.003 | 0.003 | 0.000 | ||||||||||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | |||||||||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | ||||||||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | |||||||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | ||||||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | |||||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | ||||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | |||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | ||||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | |||||
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0.190 | 0.003 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | ||||
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0.184 | 0.006 | 0.006 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | 0.009 | |||
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0.196 | 0.054 | 0.054 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | 0.057 | ||
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0.190 | 0.063 | 0.063 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.066 | 0.070 | 0.079 |
1 | Forcipular coxosternal margin with blunt, rounded or slightly flattened, hyaline lobes; tarsungulum very short |
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– | Forcipular coxosternal margin without hyaline lobes; tarsungulum moderate or long |
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3 | Trigonal sutures present on the posterior part of sternites. Tarsus of most legs bipartite |
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– | Sternal trigonal sutures absent. Tarsus of most legs a single article |
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5 | Ultimate legs with saw teeth present from prefemur to tarsus 2, saw teeth formula: 28-30-14-17-17 |
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– | Ultimate legs with saw teeth present on tibia and tarsus 1 only |
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7 | T1 with transverse suture only |
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– | T1 with transfer and other sutures |
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9 | Head without paramedian sutures; length: 19 mm, antennae short, 3+3 saw teeth on tibia and tarsus of ultimate legs |
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– | Head with incomplete paramedian sutures on the posterior half and the anteriormost quarter of the cephalic plate; length: 28–29 mm; antennae long, 4+9 saw teeth on tibia and tarsus 1 of ultimate leg |
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11 | T1 with inverted Y-shaped sutures |
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– | T1 with transverse and/or paramedian sutures |
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13 | T1 with transverse suture and two paramedian sutures; prefemur and femur of ultimate legs with dorsodistal spinous process; small species, ca 15 mm, cave in India |
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– | T1 with transverse suture and U-shaped or cruciform suture; prefemur and femur of ultimate legs without dorsodistal spinous process; caves in Europe |
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15 | T1 with transverse and cruciform sutures; head with 2 complete paramedian sutures, large species |
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– | T1 with transverse suture and characteristic U-shaped suture attached to it; head with incomplete paramedian sutures |
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17 | Labrum tridentate |
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– | Labrum unidentate |
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19 | Antennae short, head plate with incomplete anterior and posterior paramedian sutures; saw teeth on tibia and tarsus in combination 13+6 |
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– | Antennae long, head plate with posterior paramedian sutures only |
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21 | Head with two incomplete posterior paramedian sutures only; anterior margin of forcipular coxosternite strongly convex and covered by spiniform setae, cave in France |
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– | Head with two incomplete short posterior paramedian sutures only; anterior margin of forcipular coxosternite slightly rounded and barely protuberant; spiniform setae missing, cave on Tenerife |
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Scolopendromorphs are strictly terrestrial and most species are found in forest leaf litter, decomposed wood, under bark of dead trees, in the soil, under stones or in caves in the temperate and tropical areas of the world. Few species are well adapted to eremic environments (
An annotated list of the troglobitic/troglophilic
Species | Distribution | Category | References |
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Greece: Crete, Acrotiri, Cave Katholiko | Troglobite? |
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Australia: Queensland, Camooweal area, Five O’Clock Cave | Troglobite |
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Cuba | Troglobite |
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Greece: Thassos Island, cave Dracotrypa | unknown | Matic and Stavropoulos (1990) | |
Brazil: known from three iron ore caves of the “Quadrilátero Ferrífero” (Iron Quadrangle) in Minas Gerais in Mariana and Itabirito municipalities | Troglophile | ||
Caves only?; Slovenia and Croatia | Verhoeff 1933 | ||
Brazil: known from four caves (Ressurgência das Areias de Água Quente, Gruta Monjolinho, Caverna Alambari de Baixo, Caverna Santana) in Iporanga, São Paulo | Troglobite | ||
India: Assam, Garo Hills, Siju Cave | Troglophile | (Silvestri 1924) | |
India: Assam, Garo Hills, Siju Cave | Troglophile | (Silvestri 1924) | |
Botswana: Diviner’s Cave (Koanaka Hills) and Dimapo Cave (Gcwihaba Hills) | Epigean/Troglophile? |
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Caves in Spain | Troglobite | Ribaut (1915) | |
Romania: Mangalia, Movile Cave | Troglobite | This paper (see also |
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Australia: known from three caves (Nurina Cave 6N-46, Burnabbie Cave, cave 6N-1327), Roe Plains | Troglobite |
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Brazil: Bahia, Campo Formoso, only known from the type locality, Toca do Gonçalo Cave | |||
Cuba | Troglobite |
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Caves in France and Italy but also found outside caves | Troglophile | ||
Syn. |
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France: Alpes-Maritimes, Gourdon, Aven du Fourchu Cave | Troglobite |
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Spain: Tenerife Island, Cueva Felipe Reventón | Troglobite |
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Several morphological characters traditionally used in centipedes taxonomy could be subject to intraspecific variation related to postembryonic development, animal life stage and ecology (
Intraspecific distance between the two sequenced
The type locality of
Map of Europe showing geographic distribution of
We are especially grateful to Serban M. Sarbu (Adjunct Faculty, California State University Chico) for calling our attention to this interesting material and for committing samples from Movile Cave for study. Stefan Baba (“Emil Racoviță” Institute of Speleology & Faculty of Biology, University of Bucharest, Romania), Dragan Antić and Dalibor Stojanović (both from University of Belgrade – Faculty of Biology) committed further specimens of
Here belong: C. camoowealensis, C. cavernicolus, C. hephaestus, C. iporangensis, C. longicornis, C. roeplainsensis, C. troglobius