Research Article |
Corresponding author: Karin Voigtländer ( karin.voigtlaender@senckenberg.de ) Academic editor: Marzio Zapparoli
© 2017 Karin Voigtländer, Etienne Iorio, Peter Decker, Jörg Spelda.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Voigtländer K, Iorio E, Decker P, Spelda J (2017) The subgenus Monotarsobius in the Iberian Peninsula with a description of a new pseudo-cryptic species from Northern Spain revealed by an integrative revision of Lithobius crassipes L. Koch, 1862 (Chilopoda, Lithobiomorpha, Lithobiidae). ZooKeys 681: 1-38. https://doi.org/10.3897/zookeys.681.12942
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The widespread European centipede species Lithobius (Monotarsobius) crassipes L. Koch, 1862 was revised using an integrative approach incorporating sequence data and morphology. The partial mitochondrial cytochrome c oxidase subunit I (COI) barcoding gene was amplified and sequenced for 21 individuals from northern Spain, France and Germany as well as for individuals of three other species of the subgenus Monotarsobius Verhoeff, 1905. The dataset was used for molecular phylogenetic analysis and genetic distance determination. In addition, Monotarsobius specimens from more than 100 localities in northern Spain, France, and Germany were morphologically investigated. Both morphological and molecular data indicate that specimens from the Navarre and Gipuzkoa provinces, northern Spain, represent a distinct pseudo-cryptic species, only differing in some minor characters from L. crassipes. The new species L. (Monotarsobius) crassipesoidessp. n. is described and compared to L. crassipes in detail using morphology and morphometric statistics for body, head, and antennae length, number of ocelli and coxal pores, as well as the starting leg for legpair spines Vmt and DaP. The Iberian and European records of L. crassipes are discussed. The subspecies L. crassipes morenoi Garcia Ruiz, 2014 from Southern Spain is elevated to species as L. morenoi stat. n. A checklist, distribution map and key to all five species of Monotarsobius of the Iberian Peninsula are presented.
COI, taxonomy, new species, pseudo-cryptic diversity, key, checklist
Currently 53 species and seven subspecies of the genus Lithobius Leach, 1814 are known from the Iberian Peninsula (
Following a field trip to Navarre province in northern Spain in 2009 (see Material and methods), we noted that specimens of L. crassipes from the area differed slightly in morphology from other European L. crassipes. Fresh and museum material of L. crassipes from several localities in northern Spain, France and Germany was then studied in order to clearly delimit the new Spanish L. crassipes-like species, with integrated use of phylogenetic data, classical morphology and morphometric statistics. Here we also elevate L. crassipes morenoi to full species status and present an annotated list and updated key to species of Monotarsobius on the Iberian Peninsula. Finally, the implications on species distribution of L. crassipes on the Iberian Peninsula as well as in other parts of Europe are discussed.
Specimen collecting and preservation. Material of Lithobius crassipesoides sp. n. was hand-collected during a collecting trip in Navarre province (Spain) in 2009. Participants and collectors (collectively referred to as FT2009) were Karin Voigtländer, Hans Reip, Norman Lindner, Desmond Kime, Helen Read, Henrik Enghoff, Paul Richards, Steve Gregory, and Per Djursvoll.
Additional material (in ethanol or mounted at slides) was borrowed from the Museum of Zoology of the University of Navarra (
Material of Lithobius crassipes from various regions in Germany in the Senckenberg Museum of Natural History Görlitz (
Sixty-four specimens of L. crassipesoides sp. n. from Spain were morphologically studied. For L. crassipes 55 specimens from 36 localities in Germany and 131 specimens from 60 localities in France were studied (see supplementary Table
Species, localities, GenBank accession numbers, and repository accession numbers for all specimens analyzed.
Species | Locality | GenBank Acc. No. | Voucher |
---|---|---|---|
Ingroup | |||
L. crassipesoides sp. n. | Spain, Navarra, Leitza, between area „Ustarleku“ and „Karobieta“ | MF123704 |
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L. crassipesoides sp. n. | Spain, Navarra, Leitza, between area „Ustarleku“ and „Karobieta“ | MF123705 |
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L. crassipesoides sp. n. | Spain, Navarra, Leitza, near road NA 4150 | MF123706 |
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L. crassipesoides sp. n. | Spain, Spain, Navarra, Leitza, near road NA 4150 | MF123707 |
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L. crassipesoides sp. n. | Spain, Navarra, Sierra de Aralar, S Errazkin | MF123708 |
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L. crassipesoides sp. n. | Spain, Gipuzkoa, Onati, Aizkorri-Aratz | MF123709 |
|
L. crassipes | Germany, Baden-Württemberg, Bad Urach | KX458777 | GBOL14972 |
L. crassipes | Germany, Bavaria, Höllental | MF123703 |
|
L. crassipes | Germany, Bavaria, Kelheim | JQ350449 | BCZSMMYR00443 |
L. crassipes | Germany, Bavaria, Klausenhöhle | KX458796 | GBOL11199 |
L. crassipes | Germany, Bavaria, Poppendorf | KX458752 | GBOL12250 |
L. crassipes | Germany, Bavaria, Eichstätt | KX458625 | GBOL14992 |
L. crassipes | Germany, Hesse, Göttingen | JQ801572.1 | |
L. crassipes | Germany, Lower Saxony, Soltau | KX458601 | GBOL11898 |
L. crassipes | Germany, Saxony-Anhalt, Bollenkopf | MF123701 |
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L. crassipes | Germany, Schleswig-Holstein, Bad Segeberg | KX458716 | GBOL12308 |
L. crassipes | France, Haut-Rhin, Sewen | KX458631 | GBOL11870 |
L. crassipes | France, Haut-Rhin, Thann | KX458674 | GBOL11887 |
L. crassipes | France, Moselle, Ballon d`Alsace | KX458683 | GBOL11895 |
L. crassipes | France, Loire-Atlantique, Le Gâvre | MF123710 |
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L. crassipes | Great Britain, Wales, Newbridge | KX458753 | GBOL11881 |
L. curtipes | Germany, Bavaria, Neuschönau | KX458647 | GBOL12396 |
L. curtipes | Germany, Bavaria, Hengersdorf | KX458722 | GBOL14991 |
L. curtipes | Germany, Saxony-Anhalt, Elbingerode | KX458653 | GBOL11899 |
L. curtipes | Germany, Saxony-Anhalt, Elbingerode | KX458595 | GBOL11875 |
L. curtipes | Great Britain, Wales, Groesfaen Woods | KX458652 | GBOL11815 |
L. austriacus | Germany, Bavaria, Grafenau | KX458587 | GBOL11194 |
L. austriacus | Germany, Bavaria, Grafenau | KX458687 | GBOL14980 |
L. austriacus | Germany, Bavaria, Karlsfeld | KX458792 | GBOL11829 |
L. austriacus | Germany, Bavaria, Spiegelau | KX458633 | GBOL12311 |
L. austriacus | Germany, Saxony, Oybin | KX458666 | GBOL12326 |
L. austriacus | Austria, Upper Austria, Sankt Wolfgang | JQ350447 | BCZSMMYR00442 |
L. forficatus | Germany, Mecklenburg-Western Pomerania, Rügen | MF123702 |
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Outgroup | |||
E. cavernicolus | Croatia | KF715050.1 | CHP-416 |
C. parisi | Germany | KU497164.1 | GBOL02712 |
S. linearis | Germany | KM491663.1 | ZFMK-TIS-15771 |
Illustrations. For scanning electron micrographs (SEM), samples were dehydrated through an ethanol series, dried in a desiccator overnight, and mounted on aluminum stubs before being sputter coated with gold-palladium. SEM images were taken using a JEOL JSM-6510LV microscope, and samples were removed from stubs and returned to alcohol upon examination.
Preserved specimens were imaged with a Leica M165 C or Leica DM5500B stereo microscope and DFC295 camera. Focus-stacked images were assembled from 10–25 source images using the software package Leica V4.5. All images of the z-stacks are available online at VIRMISCO (www.virmisco.org,
All figures were later edited using Adobe Photoshop CS4. Maps were created with ArcGIS 10.
DNA extraction and molecular analysis. At the
Polymerase chain reaction (PCR) was used for amplifying the COI barcode fragment using the primer pair LCO1490 and HCO2198 (
As part of the GBOL project at
All 32 new sequences were deposited in GenBank (see Table
Alignment and phylogenetic analysis. All obtained sequences were checked via Blast searches (
Statistical analysis. The software package PAST version 3.14 (
Box and jitter plots: For each sample, the 25–75 percent quartiles are drawn using a box. The median is shown with a horizontal line inside the box. The minimal and maximal values are shown with short horizontal lines (“whiskers”). Each value is plotted as a dot. To show overlapping points more clearly, they have been displaced using a random “jitter” value controlled by a slider.
Usually nonparametric statistics have been used, while parametric statistics have been restricted to interval data. Prior to the parametric t-test normality was tested using three statistical tests for normal distribution: The Shapiro-Wilk test (
In the xy graph of the canonical variate analysis (Figs
The following characters have only been investigated in L. crassipesoides sp. n. from Spain and L. crassipes from Germany: the head length, the number of coxal pores on legpair 12–15 and the start of the ventral median spine on the trochanter.
Terminology. The terminology of external morphology follows
The coloration as seen in alcohol material under a stereo microscope follows the colour terminology of
The following abbreviations are used in the text and figures:
FT2009 participants in the Navarre field trip in 2009
AL antennae length
BL body length
HL head length
LP legpair
C coxa
P prefemur
F femur
T tibia
t trochanter
Ts tarsus
a anterior spine
m median spine
p posterior spine
V ventral
D dorsal
The monophyly of both Lithobius crassipesoides sp. n. and L. crassipes is well supported with bootstrap values of 99 and 98 respectively (Fig.
Statistics of body length. A comparison of jitter box plots shows that the specimens of L. crassipesoides sp. n. from Spain are usually smaller than those of L. crassipes (Fig.
Body length (mm) of males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany and France).
Lithobius crassipesoides | Lithobius crassipes | ||||||||
---|---|---|---|---|---|---|---|---|---|
Spain | France | Germany | |||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 62 | 69 | 131 | 27 | 25 | 52 |
Min | 6.94 | 6.39 | 6.39 | 8.30 | 7.90 | 7.90 | 9.40 | 7.62 | 7.62 |
Max | 11.02 | 10.74 | 11.02 | 13.20 | 13.10 | 13.20 | 12.40 | 12.10 | 12.40 |
Median | 8.47 | 8.02 | 8.43 | 9.80 | 9.90 | 9.90 | 10.60 | 9.93 | 10.20 |
Mean | 8.68 | 8.18 | 8.42 | 9.92 | 10.15 | 10.05 | 10.63 | 10.11 | 10.38 |
Stand. dev | 1.04 | 1.04 | 1.06 | 1.06 | 1.31 | 1.20 | 0.90 | 1.12 | 1.04 |
The performed tests for normality (Table
Normality test for body length (mm) of males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany and France). The p values below 0.05 are marked with bold letters.
Lithobius crassipesoides | Lithobius crassipes | ||||||||
---|---|---|---|---|---|---|---|---|---|
Spain | France | Germany | |||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 62 | 69 | 131 | 27 | 25 | 52 |
Shapiro-Wilk W | |||||||||
p(normal) | 0.387 | 0.729 | 0.690 | 0.015 | 0.015 | 0.000 | 0.089 | 0.467 | 0.416 |
Anderson-Darling A | |||||||||
p(normal) | 0.251 | 0.740 | 0.671 | 0.157 | 0.021 | 0.002 | 0.165 | 0.313 | 0.274 |
p(MCarlo) | 0.261 | 0.761 | 0.687 | 0.165 | 0.020 | 0.002 | 0.167 | 0.319 | 0.289 |
Jarque-Bera JB | |||||||||
p(normal) | 0.794 | 0.718 | 0.637 | 0.019 | 0.146 | 0.010 | 0.405 | 0.796 | 0.897 |
p(MCarlo) | 0.759 | 0.637 | 0.559 | 0.023 | 0.086 | 0.018 | 0.188 | 0.754 | 0.888 |
Using the Mann-Whitney-U-Test for mean, no significant differences in body length have been found between males and females of L. crassipesoides sp. n. from Spain (p=0.108) and between L. crassipes from France (0.433) and Germany (p=0.067). Therefore, it was justified to pool the samples. These pooled samples showed, that specimens of L. crassipesoides sp. n. are significantly shorter than specimens of L. crassipes from France (p<0.001) and Germany (p<0.001), while the two samples of L. crassipes do not significantly differ (p=0.081).
A comparison of jitter box plots shows, that the specimens of L. crassipesoides sp. n. from Spain usually have shorter heads than those of L. crassipes from Germany (Fig.
Basic statistics of head length (in mm) in males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany).
Lithobius crassipesoides | Lithobius crassipes | |||||
---|---|---|---|---|---|---|
Spain | Germany | |||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 25 | 27 | 52 |
Min | 0.72 | 0.6 | 0.6 | 0.8 | 0.8 | 0.8 |
Max | 1.08 | 1.08 | 1.08 | 1.08 | 1.12 | 1.12 |
Median | 0.86 | 0.8 | 0.84 | 0.92 | 0.9 | 0.92 |
Mean | 0.85 | 0.81 | 0.83 | 0.93 | 0.92 | 0.92 |
Stand. dev | 0.09 | 0.11 | 0.10 | 0.07 | 0.09 | 0.08 |
The performed tests for normality (Table
Normality test for head length (in mm) of males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany). The p values below 0.05 are marked with bold letters.
Lithobius crassipesoides | Lithobius crassipes | |||||
---|---|---|---|---|---|---|
Spain | Germany | |||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 27 | 25 | 52 |
Shapiro-Wilk W | ||||||
p(normal) | 0.105 | 0.572 | 0.241 | 0.040 | 0.022 | 0.002 |
Anderson-Darling A | ||||||
p(normal) | 0.080 | 0.314 | 0.101 | 0.016 | 0.037 | 0.001 |
p(MCarlo) | 0.076 | 0.316 | 0.100 | 0.015 | 0.035 | 0.001 |
Jarque-Bera JB | ||||||
p(normal) | 0.585 | 0.705 | 0.675 | 0.563 | 0.236 | 0.130 |
p(MCarlo) | 0.406 | 0.625 | 0.616 | 0.380 | 0.089 | 0.068 |
Using the Mann-Whitney-U-Test, no significant differences in head length have been found between males and females of L. crassipesoides sp. n. (p=0.105) and L. crassipes from Germany (0.655). The pooled samples show that head length was significantly shorter in specimens of L. crassipesoides sp. n. than in specimens of L. crassipes from Germany (p<0.001).
Statistics of antennae length. A comparison of jitter box plots shows, that the antennae of the specimens of L. crassipesoides sp. n. from Spain are usually smaller than those of L. crassipes (Fig.
Antennae length (mm) of males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany and France).
Lithobius crassipesoides | Lithobius crassipes | ||||||||
Spain | France | Germany | |||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 59 | 59 | 118 | 25 | 27 | 52 |
Min | 1.60 | 1.60 | 1.60 | 2.00 | 2.10 | 2.00 | 2.44 | 2.40 | 2.40 |
Max | 3.20 | 3.00 | 3.20 | 3.70 | 4.00 | 4.00 | 3.68 | 3.64 | 3.68 |
Median | 2.28 | 2.04 | 2.16 | 3.00 | 3.00 | 3.00 | 2.88 | 3.00 | 2.98 |
Mean | 2.27 | 2.05 | 2.16 | 3.01 | 2.98 | 3.00 | 2.94 | 2.99 | 2.97 |
Stand. dev | 0.39 | 0.36 | 0.38 | 0.36 | 0.42 | 0.39 | 0.34 | 0.33 | 0.33 |
The performed tests for normality (Table
Normality test for antennal length (mm) of males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany and France). The p values below 0.05 are marked with bold letters.
Lithobius crassipesoides | Lithobius crassipes | ||||||||
---|---|---|---|---|---|---|---|---|---|
Spain | France | Germany | |||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 62 | 69 | 131 | 27 | 25 | 52 |
Shapiro-Wilk W | |||||||||
p(normal) | 0.486 | 0.079 | 0.034 | 0.083 | 0.700 | 0.195 | 0.307 | 0.869 | 0.327 |
Anderson-Darling A | |||||||||
p(normal) | 0.421 | 0.200 | 0.119 | 0.053 | 0.538 | 0.043 | 0.369 | 0.891 | 0.621 |
p(MCarlo) | 0.424 | 0.203 | 0.116 | 0.054 | 0.554 | 0.042 | 0.378 | 0.896 | 0.645 |
Jarque-Bera JB | |||||||||
p(normal) | 0.726 | 0.299 | 0.291 | 0.253 | 0.932 | 0.804 | 0.739 | 0.737 | 0.569 |
p(MCarlo) | 0.645 | 0.113 | 0.156 | 0.132 | 0.931 | 0.785 | 0.668 | 0.677 | 0.466 |
Using the Mann-Whitney-U-Test, only low significant differences in antennal length have been found between males and females of L. crassipesoides sp. n. from Spain (p=0.046) but not between L. crassipes from France (0.655) and Germany (p=0.555). The very low significance in specimens of L. crassipesoides sp. n. solely is interpreted as a stochastic effect and as there are no sex-specific differences in both of the L. crassipes samples they are pooled for comparison of the two species. These pooled samples showed that the antennae of specimens of L. crassipesoides sp. n. are significantly shorter than in specimens of L. crassipes from France (p<0.001) and Germany (p<0.001, while the two samples of L. crassipes do not significantly differ (p=0.632).
Statistics of number of ocelli. The number of ocelli was counted on each side of the head separately. For statistical evaluation, the average of the two sides has been used, as the separate measures are not independent samples (
A comparison of jitter box plots shows that the specimens of L. crassipesoides sp. n. from Spain usually have a lower number of ocelli than specimens of L. crassipes from France and Germany (Fig.
Number of ocelli of males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany and France).
Lithobius crassipesoides | Lithobius crassipes | ||||||||
---|---|---|---|---|---|---|---|---|---|
Spain | France | Germany | |||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 23 | 24 | 47 | 59 | 66 | 125 | 25 | 27 | 52 |
Min | 7.5 | 5.5 | 5.5 | 7 | 8.5 | 7 | 8.5 | 8 | 8 |
Max | 11 | 10 | 11 | 14.5 | 13.5 | 14.5 | 11 | 11.5 | 11.5 |
Median | 9 | 8.5 | 9 | 10 | 10 | 10 | 10 | 10 | 10 |
Mean | 9.1 | 8.3 | 8.7 | 10.1 | 10.2 | 10.1 | 10.1 | 9.7 | 9.9 |
Stand. dev | 1.2 | 1.2 | 1.3 | 1.0 | 1.0 | 1.0 | 0.7 | 0.8 | 0.8 |
Using the Mann-Whitney-U-Test, no significant differences in the number of ocelli have been found between males and females of L. crassipesoides sp. n. from Spain (p=0.063) and L. crassipes from France (0.438) and Germany (p=0.074). The pooled samples show that the number of ocelli in specimens of L. crassipesoides sp. n. is significantly lower than in specimens of L. crassipes from France (p<0.001) and Germany (p<0.001, while the two samples of L. crassipes do not significantly differ (p=0.293).
Statistics of number of coxal pores. A comparison of jitter box plots shows that specimens of L. crassipesoides sp. n. usually have a lower number of coxal pores on legpair 12 to 15 and that females usually have a higher number of coxal pores on them (Fig.
Basic statistics of number of coxal pores on legpair 12–15 in males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany).
Lithobius crassipesoides | Lithobius crassipes | |||||
---|---|---|---|---|---|---|
Spain | Germany | |||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
legpair 12 | ||||||
N | 24 | 26 | 50 | 25 | 27 | 52 |
Min | 2 | 2 | 2 | 2 | 2 | 2 |
Max | 2 | 3 | 3 | 3 | 4 | 4 |
Median | 2 | 2 | 2 | 2 | 2 | 2 |
Mean | 2.00 | 2.08 | 2.04 | 2.12 | 2.41 | 2.27 |
Stand. dev | 0.00 | 0.27 | 0.20 | 0.33 | 0.57 | 0.49 |
legpair 13 | ||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 25 | 27 | 52 |
Min | 2 | 3 | 2 | 3 | 3 | 3 |
Max | 4 | 4 | 4 | 4 | 5 | 5 |
Median | 3 | 3 | 3 | 3 | 3 | 3 |
Mean | 3.00 | 3.12 | 3.06 | 3.08 | 3.48 | 3.29 |
Stand. dev | 0.29 | 0.33 | 0.31 | 0.28 | 0.58 | 0.50 |
legpair 14 | ||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 25 | 27 | 52 |
Min | 2 | 3 | 2 | 3 | 3 | 3 |
Max | 4 | 5 | 5 | 4 | 5 | 5 |
Median | 3 | 3 | 3 | 3 | 4 | 3 |
Mean | 3.00 | 3.42 | 3.22 | 3.20 | 3.74 | 3.48 |
Stand. dev | 0.29 | 0.58 | 0.51 | 0.41 | 0.53 | 0.54 |
legpair 15 | ||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 25 | 27 | 52 |
Min | 2 | 2 | 2 | 2 | 2 | 2 |
Max | 3 | 4 | 4 | 3 | 4 | 4 |
Median | 2 | 3 | 3 | 3 | 3 | 3 |
Mean | 2.08 | 3.00 | 2.56 | 2.60 | 3.22 | 2.92 |
Stand. dev | 0.28 | 0.40 | 0.58 | 0.50 | 0.51 | 0.59 |
In Table
Significance levels (p values) of differences in the numbers of coxal pores between males and females and pooled samples of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany). The p values below 0.05 are marked with bold letters.
legpair 12 | legpair 13 | legpair 14 | legpair 15 | |
---|---|---|---|---|
♂♂ (Spain) vs. ♀♀ (Spain) | 0.179 | 0.197 | 0.002 | < 0.001 |
♂♂ (Germany) vs. ♀♀ (Germany) | 0.037 | 0.003 | < 0.001 | < 0.001 |
♂♂ (Spain) vs. ♂♂ (Germany) | 0.087 | 0.343 | 0.06 | < 0.001 |
♀♀ (Spain) vs. ♀♀ (Germany) | 0.01 | 0.008 | 0.041 | 0.083 |
♂+♀ (Spain) vs. ♂+♀ (Germany) | 0.002 | 0.007 | 0.014 | 0.002 |
Statistics of legpair DaP spine. The start of the anterior dorsal spine (DaP) at the prefemur has sometimes been checked on both sides, sometimes on one side solely, depending on the availability of intact specimens with at least one leg being complete on one side at a given legpair position. Again, the average value has been used for the statistical calculation if both values had been available. Due to the high number of specimens with missing legs the number of samples was much lower than in the previously processed characters, see Table
Legpair where anterior dorsal spine of prefemur (DaP) starts in males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany and France). Abbreviation: n. p. = not present.
Lithobius crassipesoides | Lithobius crassipes | ||||||||
---|---|---|---|---|---|---|---|---|---|
Spain | France | Germany | |||||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 36 | 34 | 70 | 25 | 26 | 51 |
Min | 14 | 12 | 12 | 7,5 | 8 | 7.5 | 7 | 8 | 7 |
Max | 15 | n. p. | n. p. | 11.5 | 11.5 | 11.5 | 11 | 13 | 13 |
Median | 15 | 15 | 15 | 10 | 9.75 | 10 | 9 | 9 | 9 |
Mean | 14.7 | 14.4 | 14.5 | 10.0 | 9.8 | 9.9 | 9.0 | 9.8 | 9.4 |
Stand. dev | 0.5 | 1.0 | 0.8 | 0.9 | 0.7 | 0.8 | 1.2 | 1.4 | 1.4 |
A comparison of jitter box plots shows that the spine DaP usually starts later in specimens of L. crassipesoides sp. n. from Spain than in specimens of L. crassipes (Fig.
Using the Mann-Whitney-U-Test, no significant differences in the starting position of spine DaP have been found between males and females of L. crassipesoides sp. n. from Spain (p=0.209) and L. crassipes from France (p=0.340), while there was a low significance between males and females from Germany (p=0.024). The pooled samples show that spine DaP starts significantly later in specimens of L. crassipesoides sp. n. than in specimens of L. crassipes from France (p<0.001) and Germany (p<0.001). However, the two samples of L. crassipes differ significantly (p=0.001).
The start of the median ventral spine at the trochanter (Vmt) has sometimes been checked on one side solely. The basic descriptive statistic parameters are given in Table
Start of the anterior dorsal spine of prefemur in males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany and France). The DaP spine has been recorded on each leg of each specimen in French material: this explains the 0.5 graduation in this case (DaP being sometimes present on one side only).
Basic statistics of legpair where median ventral trochanter spine (Vmt) starts in males and females of Lithobius crassipesoides sp. n. (Spain) and L. crassipes (Germany).
Lithobius crassipesoides | Lithobius crassipes | |||||
---|---|---|---|---|---|---|
Spain | Germany | |||||
♂♂ | ♀♀ | ♂+♀ | ♂♂ | ♀♀ | ♂+♀ | |
N | 24 | 26 | 50 | 25 | 27 | 52 |
Min | 12 | 12 | 12 | 13 | 13 | 13 |
Max | 14 | 14 | 14 | 14 | 14 | 14 |
Median | 13 | 13 | 13 | 14 | 14 | 14 |
Mean | 13.0 | 13.0 | 13.0 | 13.9 | 13.9 | 13.9 |
Stand. dev | 0.6 | 0.3 | 0.5 | 0.3 | 0.4 | 0.3 |
A comparison of jitter box plots shows that the spine Vmt usually starts earlier in specimens of L. crassipesoides sp. n. from Spain than in specimens of L. crassipes (Fig.
Using the Mann-Whitney-U-Test, no significant differences in the starting position of spine Vmt have been found between males and females of L. crassipesoides sp. n. (p=0.777) and L. crassipes from Germany (p=0.771). The pooled samples show that spine Vmt appeared significantly earlier in specimens of L. crassipesoides sp. n. than in specimens of L. crassipes from Germany (p<0.001).
A canonical variate analysis (CVA, linear discriminant analysis with several groups) has been performed to improve the separation of the two species. As complete datasets are necessary for such a comparison only samples from Germany could be used to represent L. crassipes. All morphometric characters evaluated above have been used and sex specific groups have been defined. Additionally, the ratios of body length and antenna length to head length have been included.
The CVA (Fig.
Although differences between the sexes are clearly visible, even CVA does not allow separating them completely. Three discriminant factors have been extracted, explaining 85.8 %, 12.1 % and 2.1 % of the observed variance.
The factor loadings of discriminant function one show that the start of spine DaP and the body length have the highest separating power for the two species (Table
Eigenvalues and factor loading of the three extracted discriminant factors for the complete model. Highest factor loadings marked in bold letters and top values in larger fonts.
Axis 1 | Axis 2 | Axis 3 | |
---|---|---|---|
Eigenvalue | 10.6 | 1.49 | 0.26 |
Explanation of variance | 85.8 % | 12.1 % | 2.1 % |
Body length | -0.303 | 0.080 | 0.579 |
Antennae length | -0.125 | 0.063 | 0.147 |
Head length | -0.014 | 0.015 | 0.015 |
AL/HL | -0.097 | 0.026 | 0.132 |
BL/AL | 0.067 | -0.071 | -0.004 |
Coxal pores LP 15 | -0.064 | -0.316 | 0.012 |
Coxal pores LP 14 | -0.044 | -0.187 | 0.157 |
Coxal pores LP 13 | -0.037 | -0.090 | 0.197 |
Coxal pores LP 12 | -0.037 | -0.062 | 0.145 |
Ocelli | -0.170 | 0.262 | 0.120 |
DaP | 0.804 | -0.202 | 0.400 |
Vmt | -0.132 | 0.017 | -0.001 |
Discriminant factor two could be interpreted as a “sex factor”, with the highest factor loading on the number of coxal pores on legpair 15. Ocelli number and start of DaP are also of importance, while the numbers of coxal pores on the other legs are redundant characters.
Factor three is of no practical value and explains only a negligible part of the variance (Fig.
Based on these interpretations a reduced model could be defined, which has a lower separating power but requires less measuring. Fig.
Eigenvalues and factor loading of the three extracted discriminant factors in a reduced model. Highest factor loadings marked in bold letters.
Axis 1 | Axis 2 | Axis 3 | |
---|---|---|---|
Eigenvalue | 9.84 | 1.23 | 0.13 |
Explanation of variance | 87.9 % | 11.0 % | 1.1 % |
Body length | -0.313 | -0.080 | 0.904 |
Antennae length | -0.130 | -0.063 | 0.235 |
Coxal pore LP15 | -0.063 | 0.352 | 0.099 |
Ocelli | -0.179 | -0.278 | 0.167 |
DaP | 0.838 | 0.145 | 0.393 |
Vmt | -0.138 | -0.008 | 0.033 |
Both molecular analysis (Fig.
The main weakness of the morphometric statistical evaluation is the fact that the measured specimens are not identical with the sequenced specimens. This means that the possibility of having other cryptic species among the material cannot be excluded. The rejection of the normal distribution in the French samples in body length, in the German samples in head length and in samples from Spain and France in antennae length might be interpreted as a hint for more cryptic lineages.
The moderate dropping of the eigenvalue of axis1 from 10.6 to 9.84 (7 %) and of axis2 from 1.49 to 1.23 (1.5 %) shows that no crucial variables have been removed when generating the reduced model. This is also visible by comparison of Fig.
Canonical variate analysis of morphometric data of Lithobius crassipesoides sp. n. from Spain and L. crassipes from Germany, showing factors one and two. Only characters with high separating power have been included. The specimen
As seen in Fig.
Currently, all records of L. crassipes on the Iberian Peninsula as well as in other parts of Europe need to be carefully checked against L. crassipesoides sp. n., which could also be dispersed by human transport (
Interestingly, some authors have previously given details on morphology of Spanish L. crassipes.
Here we give a brief review of historic descriptions of L. crassipes for characters which have been found to be useful. Descriptions of L. crassipes elsewhere in Western Europe quoted a body length of 9–12 mm (
In Sweden, western specimens of L. crassipes seems to differ from the south-eastern Swedish L. crassipes (including also those from Denmark) as underlined by
Species of the subgenus Monotarsobius are characterized by short antennae and, in its modern conception, very frequently with 20 antennal articles for most members in Western Europe; the wider amplitude in this area being usually 17–23 articles, very rarely 24–25 in exception as in Spanish L. blascoi Eason, 1991. Monotarsobius always has 2 + 2 forcipular teeth, no posterior projections on tergites and tarsal articulation is missing on the first 12 or 13 legpairs (
Lithobius
crassipes
–
Small member (body length 6.4–11 mm) of the subgenus Monotarsobius. Antennae with 20 articles, short, 2.6 times longer than head, 1/4 of body length. 5–11 ocelli, mostly 8 or 9, in two or three rows with one larger posterior ocellus. Legs with species-specific plectrotaxy; legpair 14 and 15 thickened in both sexes, much more so in males; legpair 15 without accessory apical claw, in males with a depression in the posterior half of tibia. Female gonopod claw tridentate.
L. crassipesoides sp. n. differs generally from other Iberian members of Monotarsobius in the presence of a depression in the posterior half of the legpair 15 tibia. It differs from L. osellai and L. morenoi in having more than one row of ocelli; from L. blascoi in having only 20 antennal articles; and from L. crassipes in smaller body length, shorter antennae, lower number of ocelli, the DaP spine starting posteriorly from legpair (12) 13, and the different location and size of the male depression on legpair 15 tibia.
Derived from the morphological similarity to Lithobius crassipes.
Holotype. Spain: Navarre province: Leitza, Ariz Mendiak, between area “Ustarleku” and “Karobieta”, above side stream to Gorriztaran; loamy and calcareous soil, 43.0778°N, 1.8775°W, 615 m a.s.l., 20 April 2009, leg. FT2009, grove of Castanea, pollard trees on the slope with Ranunculus ficaria, Daphne, Helleborus, Salvia, Rubus, Lathrea; under leaves and bark of rotten trunk, 1 ♂ (
Paratype. Spain: Navarre province: Same data as holotype, 1 ♂ (
Other material examined.Spain: Navarre province: Leitza, Ariz Mendiak, between area “Ustarleku” and “Karobieta”, above side stream to Gorriztaran; loamy and calcareous soil, 43.0778°N, 1.8775°W, 615 m a.s.l., 20 April 2009, leg. FT2009, grove of Castanea, pollard trees on the slope with Ranunculus ficaria, Daphne, Helleborus, Salvia, Rubus, Lathrea; under leaves and bark of rotten trunk, 2 ♂♂, 3 ♀♀, 1 juv. ♂ (
Habitus. Slightly fusiform, widest around tergite 10 (Fig.
Colour. General body colouration varies from pale and light buff (most individuals), through yellow ochre to tawny olive (Fig.
Length. 6.4–11.0 mm (Fig.
Head. Head roundish, mostly as broad as long or little broader than long and head broader or as broad as T5. Head length 0.6–1.08 mm (Fig.
Antennae. 20 antennal articles, short and stout, 1.6–3.2 mm long, 2.6 times longer than head (Figs
Ocelli. 5 to 11 ocelli on each side of the head, mostly 8 or 9, rarely 5 or 11, arranged in two or three rows (Fig.
Coxosternum. Anterior border with 2+2 teeth, upper part slender, acuminate, lateral borders without shoulders. Middle notch narrow to moderate width. Sometimes coxosternum slenderer and the middle notch narrower (Fig.
Tergites. Surface slightly rough, glossy. Posterior border of T1 feebly concave or straight, T3 to T5 feebly concave, T8 to T15 distinctly concave, T16 feebly to distinctly concave. Posterior angles of T9, T11 and T13 mostly obtuse or rounded with no trace of lobes or triangular projections.
Legs. Tarsus and metatarsus fused on legpair 1 to 11. On legpair 12 and 13 the tarsal-metatarsal articulation is indistinct. Penultimate and ultimate legpairs (14, 15) are densely covered with pores. Last two legpairs are thickened in both sexes, much more so in males. Without accessory apical claw on legpair 15.
Legpair 15 tibia of males with a more setiferous depression (fossa), which is distinct and well-developed in specimens in later developmental stages (Fig.
Coxal pores. Round, 2–4 (sometimes 5) pores on each coxa (Fig.
Plectrotaxy. The plectrotaxy of legs of L. crassipesoides sp. n. is given in Table
Plectrotaxy of Lithobius crassipesoides sp. n. (n = 11 ♂♂ and 11 ♀♀). In brackets spines absent in more than 50 % of individuals.
Legpair | Ventral | Dorsal | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
C | t | P | F | T | C | t | P | F | T | |
1 | – | – | (p) | am(p) | m | – | – | m p | a(p) | a |
2 | – | – | (p) | am(p) | (a)m | – | – | m p | a p | a(p) |
3 | – | – | (p) | am(p) | (a)m | – | – | m p | a p | a(p) |
4 | – | – | (p) | am(p) | (a)m | – | – | m p | a p | a(p) |
5 | – | – | (p) | am(p) | (a)m | – | – | m p | a p | a(p) |
6 | – | – | (p) | am(p) | (a)m | – | – | m p | a p | a(p) |
7 | – | – | (p) | am(p) | (a)m | – | – | m p | a p | a(p) |
8 | – | – | (p) | am(p) | a m | – | – | m p | a p | a(p) |
9 | – | – | (p) | a m p | a m | – | – | m p | a p | a p |
10 | – | – | (mp) | a m p | a m | – | – | m p | (a)p | a p |
11 | – | – | (m)p | a m p | a m | – | – | m p | (a)p | a p |
12 | – | (m) | (a)mp | a m p | a m | (a) | – | (a)mp | p | (a)p |
13 | – | m | a m p | a m p | a m | a | – | (a)mp | p | p |
14 | – | m | a m p | a m p | (a)m | a | – | (a)mp | p | (p) |
15 | – | m | a m p | am(p) | – | a | – | a m p | (p) | – |
Male gonopods. Uni-articulated.
Female gonopods. Basal article with two conical spurs on each side, their apical edge serrated (Fig.
So far only known from Navarre and Gipuzkoa provinces, northern Spain (Fig.
L. crassipes was recorded from the Navarre region from Irati (
Lithobius
crassipes
L.
Lithobius
crassipes
–
Habitus. Slightly fusiform, widest around tergite 10 (Fig.
Colour. General body colouration varies from pale horn colour to cream colour and to yellow ochre (Fig.
Length. 7.6–13.2 mm (Fig.
Head. Head roundish, mostly as broad as long or little broader than long and head broader or as broad as T5. Head length 0.8–1.12 mm (Fig.
Antennae. 20 antennal articles, short and stout, 2–4 mm long, 3.2 times longer than head, length (Figs
Ocelli. The German specimens have 8–12, the French 7–15 ocelli on each side of the head, mostly 9–11 (Fig.
Coxosternum. Anterior border with 2+2 teeth, upper part slender, acuminate, lateral borders without shoulders. Middle notch narrow to moderate width. Sometimes coxosternum broader and the middle notch broader (Fig.
Tergites. Surface slightly rough, glossy. Posterior border of T1 feebly concave or straight, T3 to T5 feebly concave, T8 to T15 distinctly concave, T16 feebly to distinctly concave. Posterior angles of T9, T11 and T13 mostly obtuse or rounded with no trace of lobes or projections.
Legs. Tarsus and metatarsus fused on legpair 1 to 11. On legpair 12 and 13 the tarsal-metatarsal articulation is indistinct. Penultimate and ultimate legpairs (14, 15) are densely covered with pores. Last two legpairs are thickened in both sexes, much more so in males. Without accessory apical claw on legpair 15.
Legpair 15 tibia of males with a more setiferous depression (fossa), which is distinct and well-developed in specimens in later developmental stages (Fig.
Coxal pores. Round, 2–4 (sometimes 5) pores on each coxa (Fig.
Coxal pores in Lithobius crassipesoides sp. n. and in German L. crassipes. Sequence from legpair 15–12.
Lithobius crassipesoides sp. n. | Lithobius crassipes (Germany) | |||
---|---|---|---|---|
♂♂ | ♀♀ | ♂♂ | ♀♀ | |
n | 31 | 31 | 26 | 26 |
Variants of arrangements | 7 | 7 | 5 | 14 |
Highest number | 2432, 2342 | 4543 | 3443 | 4554 |
Common arrangements | 2332 (71 %) | 3332 (45 %) | 2332 (42 %) | 2332 (15 %) |
2332 (32 %) | 3332 (39 %) | 3443 (15 %) |
Plectrotaxy. The plectrotaxy of legs of L. crassipes is given in Table
Plectrotaxy of German Lithobius crassipes (n = 11 ♂♂ and 11 ♀♀). In brackets spines absent in more than 50 % of individuals.
Legpair | Ventral | Dorsal | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
C | t | P | F | T | C | t | P | F | T | |
1 | – | – | – | a m | m | – | – | m p | a(p) | a |
2 | – | – | – | a m | (a)m | – | – | m p | a p | a(p) |
3 | – | – | – | a m | (a)m | – | – | m p | a p | a(p) |
4 | – | – | – | am(p) | (a)m | – | – | m p | a p | a p |
5 | – | – | – | am(p) | (a)m | – | – | m p | a p | a p |
6 | – | – | – | am(p) | a m | – | – | m p | a p | a p |
7 | – | – | – | am(p) | a m | – | – | (a)mp | a p | a p |
8 | – | – | – | am(p) | a m | – | – | (a)mp | a p | a p |
9 | – | – | (mp) | a m p | a m | – | – | a m p | a p | a p |
10 | – | – | m(p) | a m p | a m | – | – | a m p | a p | a p |
11 | – | – | m p | a m p | a m | – | – | a m p | a p | a p |
12 | – | – | (a)mp | a m p | a m | (a) | – | a m p | p | p |
13 | – | (m) | a m p | a m p | a m | a | – | a m p | p | p |
14 | – | m | a m p | a m p | m | a | – | a m p | p | (p) |
15 | – | m | a m p | a m | – | a | – | a m p | (p) | – |
Male gonopods. Uni-articulated.
Female gonopods. Basal article with two conical spurs on each side, their apical edge serrated (Fig.
Widespread in Europe. The species has been recorded from several more or less precise Spanish localities (Fig.
All current records of L. crassipes on the Iberian Peninsula are doubtful and need further verification (see discussion above). Some records from the literature have been assigned here to L. crassipesoides sp. n. or L. morenoi.
Lithobius crassipes morenoi Garcia Ruiz and Baena, 2014: 58–61, fig. 1, tables I–II.
Lithobius
crassipes
morenoi
– Serrra 1982: (record). – Garcia Ruiz and Baena 2014: 61 (discussion). –
Known from two caves (Sima LQ-14, Abuchite, Luque; Sima de la Sierrezuela/Sima Fuente del Francés, Carcabuey) in Andalusia and from Jerez de la Frontera (Cádiz), Spain (Fig.
This subspecies was described by Garcia Ruiz and Baena (2014) based on specimens from two caves in the Cordoba province in Andalusia. It clearly differs from L. crassipes crassipes in the low number of ocelli (only three), the enlarged Tömösváry’s organ, and the absence of some spines, particularly of Vmt, VaF, and DaH on legpair 15. Garcia Ruiz and Baena (2014) also discuss the record of L. crassipes by
Lithobius
blascoi
Lithobius
blascoi
– Garcia Ruiz and Baena 2014: 56 (listed). –
So far only known from the type locality Pina de Ebro near Zaragoza, Spain (Fig.
Lithobius osellai Matic 1968: 123–125, fig. 4
Lithobius
osellai
– Serrra 1982: 47 (record, description), 49 (in key). – Garcia Ruiz and Baena 2014: 56 (listed). –
Lithobius
cf.
osellai
–
In Spain (Fig.
Key is valid for adult specimens only.
1 | 3 or 4 ocelli arranged in one row only | 2 |
– | 6–13 ocelli arranged in two or three rows | 3 |
2 | Ventral plectrotaxy of LP15 = --, m, amp, -m-, ---. VaP present from LP13 to LP15, VpF from LP8 to LP14, VaT from LP7 to LP13 and DaP from LP10 to LP15. Male LP15 tibia with a weak depression on the dorsal side | L. morenoi Garcia Ruiz & Baena, 2014 stat. n. |
– | Ventral plectrotaxy of LP15 = --, m, -mp, -m-, -m-. No VaP, VpF, VaT and DaP spines on legs | L. osellai Matic, 1968 |
3 | Antennae with 22 to 23 (rarely 24, unusually 25) articles. 6–7 ocelli arranged in two rows (usually 1 + 3, 3). Angular shoulders on each side of the lateral teeth on the forcipular coxosternite. No VaP, VaF, VpF, DaP nor DpT spines on legs. Male with a dorsal wart near the extremity of the LP15 femur and a weak sulcus in the distal two thirds of LP15 tibia | L. blascoi Eason, 1991 |
– | Antennae with usually 20 articles (18–21). 6–13 ocelli arranged in two or three rows. No angular shoulders on each side of the lateral forcipular teeth. VaP at least exist on LP15, VaF at least from LP12 to LP15, VpF at least from LP13 to LP14, DaP at least on LP15 (or very rarely lacking), DpT at least on LP12 and/or LP13. Male with a depression on dorsal side of LP15 tibia | 4 |
4 | Body usually 6–11 mm long. Usually, DaP starts on LP14 or on LP15, exceptionally from LP12, rarely from LP13 or totally lacking. Male dorsal depression of LP15 tibia shorter; starts in the posterior half of LP15 tibia (Fig. |
L. crassipesoides Voigtländer, Iorio, Decker & Spelda, sp. n. |
– | Body 8.5–12 mm long (extreme values 7.9 and 13.2). Usually, DaP starts from LP7 to LP11 until LP15, very rarely from LP12 or LP13. Male dorsal depression of LP15 tibia longer, starts in the first third of LP15 tibia (Fig. |
L. crassipes L. Koch, 1862 |
We thank Ulrich Burkhardt for support in the lab, Birgit Lang on the SEM and Diana Goernert taking z-stacks on stereo microscopes (all
Jerome Moriniere (
This is a publication supported of the German Barcode of Life (GBOL) project of the Humboldt Ring, financed by the German Federal Ministry for Education and Research (FKZ 01LI1101A and FKZ 01LI1101B).
The publication of this article was funded by the Open Access Fund of the Leibniz Association.
Full data of investigated specimens of Lithobius crassipes
Data type: Occurence and collection
Explanation note: Information on locality, date, collector, collection, sex of investigated specimens of Lithobius crassipes.