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Accepted by M. Pellinen: 2 Jul. 2015; published: 11 Aug. 2015
ZOOTAXA
ISSN 1175-5326 (print edition)
ISSN 1175-5334 (online edition)Copyright © 2015 Magnolia Press
Zootaxa 3999 (3): 401–412
www.mapress.com/zootaxa/
Article
401
http://dx.doi.org/10.11646/zootaxa.3999.3.5
http://zoobank.org/urn:lsid:zoobank.org:pub:FC57F570-73DF-4C9C-80B9-4AA1D9725656
Taxonomy 2.0: Sequencing of old type specimens supports the description of two
new species of the Lasiocampa decolorata group from Morocco
(Lepidoptera, Lasiocampidae)
WOLFGANG SPEIDEL1
, AXEL HAUSMANN2
, GÜNTER C. MÜLLER3
, VASILIY KRAVCHENKO4
,
JOSEF MOOSER5
, THOMAS J. WITT6
, KHALID KHALLAAYOUNE7
,
SEAN PROSSER8
& PAUL D.N. HEBERT8
1
Zoologische Staatssammlung München, Münchhausenstraße 21, D-81247 München and Museum Witt, Tengstr. 33, D-80796
München, Germany. E-mail: [email protected]
2
SNSB, Zoologische Staatssammlung München, Münchhausenstraße 21, D-81247 München, Germany.
E-mail: [email protected]
3
Department of Parasitology, Kuvin Centre for the Study of Infectious and Tropical Diseases, Hadassah-Medical School, Hebrew
University, Jerusalem, Israel. E-mail: [email protected]
4
Department of Zoology, Tel Aviv University, Tel Aviv, Israel. E-mail: [email protected]
5
Seilerbruecklstr. 23, D-85354 Freising, Germany. E-mail: [email protected]
6
Museum Witt, Tengstr. 33, D-80796 München, E-mail: [email protected]
7
Unité de Parasitologie et des Maladies Parasitaires, Département de Pathologie et de Santé Publique Vétérinaires. Institut
Agronomique et Vétérinaire Hassan II, BP 6202, Rabat-Instituts 10100, Morocco. E-mail: [email protected]
8
Biodiversity Institute of Ontario, University of Guelph, Guelph, Canada.
E-mail: [email protected]; [email protected]
Abstract
The type of Lasiocampa decolorata (KLUG, 1830), collected in 1820, was successfully barcoded to generate a 658bp
COI-fragment after 194 years. The resulting molecular data allowed the description of two closely related species from
Morocco: Lasiocampa hannae SPEIDEL, MOOSER & WITT sp. n. from the Anti Atlas and Lasiocampa editae SPEI-
DEL, MOOSER & WITT sp. n. from the High Atlas.
Key words: DNA barcode, Lasiocampa decolorata, Lasiocampa hannae, Lasiocampa editae, Lasiocampa staudingeri,
new species, taxonomy, Morocco, Egypt, North Africa
Introduction
The use of sequence data from a standardized region in the mitochondrial genome (COI 5’, ‘DNA barcode region’)
has been proposed as a tool for species identification (HEBERT et al. 2003) and for descriptive (alpha-) taxonomy
(e.g. HEBERT et al. 2004). After controversial discussions about the suitability of such molecular data for
taxonomy (cf. e.g. TAUTZ et al. 2002, 2003, WILL et al. 2005), there is now broad acceptance of the “integrated
taxonomic approach” which combines molecular and morphological data (e.g. TELETCHEA 2010; PADIAL et al.
2010, GOLDSTEIN & DeSALLE 2011, HAUSMANN 2011). Although this approach, recently dubbed ‘taxonomy
2.0’ (JÖRGER & SCHRÖDL 2014), is now widely accepted, the crucial point, i.e. the correct linkage of molecular
data with type specimens as the name-bearing key vouchers is still infrequent. Following initial success in
sequencing old specimens with rather demanding, time-consuming methods (HAUSMANN et al. 2009a, 2009b), a
different approach which focused on the recovery of a short 164bp amplicon from the centre of the COI 5’ barcode
gene fragment yielded a very high success rate (80%) for thousands of geometrid type specimens. Here we report a
recent breakthrough in recovering the full 658bp COI barcode fragment from a 194 year old type specimen
(Lasiocampa decolorata) in one single step through next-generation sequencing (NGS).
SPEIDEL ET AL. 402 · Zootaxa 3999 (3) © 2015 Magnolia Press
The poorly known Lasiocampa decolorata (KLUG, 1830) is rarely mentioned in modern literature
(LEWANDOWSKI & FISCHER 2005, 2008, 2012), and the few existing reports have only led to small advances
in the knowledge of this enigmatic species. Initially described in 1830 from Lower Egypt (near Alexandria,
erroneously indicated as ‘Upper Egypt’ in De FREINA & WITT 1987), several populations of this species were
discovered a few decades ago in Algeria and Tunisia, as well as a single questionable record from Morocco (De
FREINA & WITT 1987). In this report, the species was exclusively found in habitats located in desert plains at
higher elevation (De FREINA & WITT 1987), contrasting with the wide spectrum of environments inhabited by
the Lasiocampa decolorata group. Indeed, the analysis of the currently known localities of the Lasiocampa
decolorata species-group with several recently detected populations in Morocco shows that it inhabits areas
ranging from about sea-level to more than 2,000 m, exhibiting an elevational range which is quite remarkable for a
single species.
The typical Lasiocampa decolorata from Egypt remained nearly unknown, and the Algerian, Tunisian and
Libyan populations that had been described under separate taxon names were never compared to the nominotypical
Egyptian population. It was essential, therefore, to obtain more information about the type specimen of
Lasiocampa decolorata which was found in an area now close to the harbour of metropolitan Alexandria
(EHRENBERG 1828, BAKER 1997, LEWANDOWSKI & FISCHER 2012), where it is probably extinct. Since
only a single female type specimen (holotype by monotypy) is known from this locality, morphological analysis
does not reliably clear up the identity of the taxon decolorata KLUG, 1830 and the identity of the Moroccan
populations because of (1) the absence of valuable differential traits in females and (2) the absence of females in
the available material from the Moroccan populations. The successful recovery of the full COI barcode fragment
from the 194 year old holotype (collected in 1820), however, finally allows an unambiguous taxonomic analysis in
this group.
Material and methods
Abbreviations
ZSM Bavarian State Collection of Zoology, Munich
CCDB Canadian Centre for DNA Barcoding
MWM Museum Witt, Munich
BOLD Barcode of Life Data Systems
BIN Barcode Index Number (Global Unique Identifier)
COI mitochondrial cytochrome c oxidase I (COI) gene, region near the 5’ terminus (barcode fragment, 658
bp)
Sampling. Some 500 Palearctic specimens of the genus Lasiocampa have been examined at the Museum
Thomas WITT (MWM) and the Bavarian State Collection of Zoology (ZSM). For identification, 50 male and
female dissections were made, using standard procedures (ROBINSON 1976).
DNA sequences for 105 Palearctic specimens in the genus Lasiocampa were available for analysis, 11
belonging to the North African 'Lasiocampa decolorata species-group'. These sequences were obtained from DNA
extracts obtained from a single leg from dried specimens in various museums and private collections. Barcode
sampling strategy aims to gain comprehensive coverage across the distribution of each species. At the present
stage, potential sampling bias due to insufficient geographical coverage is expected to play a negligible role.
DNA Analysis. PCR amplification and DNA sequencing was performed at the CCDB, for most specimens,
following standard high-throughput protocols (IVANOVA et al. 2006; DeWAARD et al. 2008) that can be
accessed under http://www.dnabarcoding.ca/pa/ge/research/protocols. PCR amplification with a single pair of
primers consistently recovered a 658 bp region near the 5’ terminus of the mitochondrial cytochrome c oxidase I
(COI) gene that included the standard 648 bp barcode region for the animal kingdom (HEBERT et al. 2003). DNA
extracts are stored at both the CCDB and in the DNA-Bank facility of the ZSM (see http://www.zsm.mwn.de/
dnabank/). Another publication (PROSSER et al. 2015) will describe lab protocols and data analysis for the 194
years old type specimen, submitted to NGS sequencing (Torrent PGM316).
Zootaxa 3999 (3) © 2015 Magnolia Press · 403LASIOCAMPA DECOLORATA
All sequences are deposited in GenBank according to the iBOL data release policy. Complete specimen data
including images, voucher deposition, GenBank accession numbers, GPS coordinates, sequence and trace files can
easily be accessed in the Barcode of Life Data System (RATNASINGHAM & HEBERT 2007) in the public
dataset DS-LASIONA.
Data analysis. Sequence divergences for the barcode region were calculated using the Kimura-2-Parameter
model, employing the analytical tools on BOLD (RATNASINGHAM & HEBERT 2007) and MEGA 6 (TAMURA
et al. 2013). Genetic distances between species are reported as minimum pairwise distances, while intraspecific
variation is reported as maximum pairwise distances.
Systematic part
Lasiocampa decolorata (KLUG, 1830)
Gastropacha decolorata KLUG, 1830, in Hemprich & Ehrenberg, Symbolae Physicae, pars Zoologica 2 (series 1 Insecta,
Decas 2): unpaginated [9], pl. 20, fig. 1. Locus typicus: ad puteos Dscheil el achterie prope Alexandriam [Egypt]
Synonyms:
Bombyx datini MABILLE, 1888, Annls Soc. ent. Fr. (6) 8 (Bull.): xlii (Locus typicus: Gabès [Tunisia]), redescribed as Bombyx
datini OBERTHÜR, 1890, Études Ent. 13: 29, pl. 6, fig. 31, 32 (Locus typicus: Gabès).
Lambessa decolorata ssp. sordidior ROTHSCHILD, 1914, Novit. zool. 21: 314. Locus typicus: Prov. Alger Central, Hauts
Plateaux (Central Algeria).
Material examined. Holotype ♀: Decolorata, N, Tscheila Ehrenb, 9018, BC ZSM Lep 81612. Museum für
Naturkunde Berlin; 1 ♂ Syntype Lambessa decolorata sordidior Type Rothsch., syntype; Guelt-es-Stel, C. Algeria,
September 1913 (V. Faroult), Rothschild Bequest 1939-1;
1 ♀ Syntype Guelt-es-Stel, Centr. Algeria (V. Faroult), 24. 9. 13, Rothschild Bequest B. M. 1939-1; 1 ♀,
Tunisia, Metameur, 33.383° N, 10.373 E, 160 m, 30. X. 2001, leg. H. Fischer, coll. J. Mooser, BC ZSM Lep 81513;
1 ♂, Algeria c., [Djelfa], Guelt-es-Stel, V.1931, Franz Daniel, München, BC ZSM Lep 80425, genitalia slide
19874. coll. MWM/ZSM.
Ehrenberg and Hemprich accompanied the famous Minutoli expedition to Egypt for the discovery of
antiquities in order to collect Natural History objects and discovered, in 1820, a single female specimen (holotype)
of this species. Under the leadership of Ehrenberg and Hemprich (EHRENBERG, 1828) some members of this
expedition left Dscheil el Achterie near Alexandria en route to Cyrenaica but it was never reached. According to
LEWANDOWSKI & FISCHER (2012) the collecting date can be reconstructed as either September 23 or 28,
1820, and according to the label, the moth was collected by Ehrenberg.
Re-description. The species was described after a female specimen. The male characters are taken from the
taxon sordidior, which the molecular genetic analysis places near L. decolorata. Male: Wing-shape and antenna-
structure corresponding to L. trifolii (DENIS & SCHIFFERMÜLLER, 1775). As in all species of the group
unicolourous yellowish brown. The Algerian populations with a greyish tinge. Female: The winged female
holotype is very pale, but this is not likely a result of bleaching because the original coloured figure already shows
a very pale moth. The population from Egypt is known only by a single specimen collected in 1820 in an area
which is now part of the metropolitan harbour area of Alexandria.
Male genitalia (genitalia slide 19874, fig. 1): No morphological description seems to exist in literature for the
male genitalia of Lasiocampa. A general, tentative description is given here for explaining the terminology of the
below given differential analyses. Male genitalia of Lasiocampa are very modified, strongly diverging from the
lepidopteran groundplan. They are situated almost on the ventral side of the abdomen, not at its caudal end as
usual. This position is caused by a slight extension of the tergal abdominal part which is bent downwards at the
end. The vinculum is quite enlarged, with two lateral processes - rolled in in some preparations. The valvae are
upwards directed in our figures, but, in natural position, they are pointing caudad. The tegumen is membranous, the
valvae consist of two lobes, the inner ones probably representing the sacculus. The inner lobes are weaker
sclerotized than the external ones and are easily damaged when the valvae are brought to a standard position, as
they are fused to some extent. The external lobes are individually quite variable in shape in all species dissected in
numbers (e.g. Lasiocampa trifolii), whereas the shape of the inner ones seems to be specific. In Lasiocampa
decolorata, the inner lobe of valva is pointed, covered with setae. Vesica dorsolaterally everted, aedeagus with a
strong, sclerotized projection at its ventral side.
Distribution. Only known with certainty from the type locality, but morphology and barcode data suggest
Tunisian and Algerian records are also attributed to this species. While records from Palestine are erroneous (see
SPEIDEL ET AL. 404 · Zootaxa 3999 (3) © 2015 Magnolia Press
discussion), further analysis should ascertain if populations from Libya and eastern lowland Morocco also
represent this species.
FIGURE 1. Lasiocampa decolorata (KLUG, 1830). Genitalia slide Heterocera Nr. 19874 MWM "Algeria c., Guelt es Stel,
[19]31"; Aedeagus Genitalia slide SP 1522 same locality. ZSM.
FIGURE 2. Lasiocampa hannae sp. n. Genitalia slide Heterocera Nr. 19873 MWM "Morocco, Anti Atlas Mts, Sidi M’zal
village, 1260 m, 29°48‘00’’ N; 8°52’11’’ W, 30. ix. 2005, leg. Gy. M. László & G. Ronkay". MWM.
FIGURE 3. Lasiocampa editae sp. n. Genitalia slide Heterocera Nr. 19872 MWM "Morocco, High Atlas, Oukaimeden, NW
slopes, 2,0–2,300 m, 11.–20. 08. 2012, G. Muller, E. Revay et al., BC ZSM Lep 76653, Museum Witt München".
FIGURE 4. Lasiocampa staudingeri (BETHUNE-BAKER, 1885). Genitalia slide Heterocera Nr. 8691 MWM – Aedeagus
mirror-inverted. “Coll. A. Seitz, Algerien, Lambessa, Coll. Th. Witt, München/Weiden”.
Zootaxa 3999 (3) © 2015 Magnolia Press · 405LASIOCAMPA DECOLORATA
SPEIDEL ET AL. 406 · Zootaxa 3999 (3) © 2015 Magnolia Press
Ecology. A photograph of the environment west of the type locality is given by LEWANDOWSKI &
FISCHER 2012 (page 101). Flight period is September.
Discussion. The species was formerly believed to occur across North Africa in scattered populations which
almost all have been described under separate subspecies or species-names, but were later regarded as synonyms.
The following additional taxa were placed in synonymy with L. decolorata by De FREINA & WITT 1987:
Lambessa virago ROTHSCHILD, 1912, Novit. zool. 19: 118 (Locus typicus: Col de Sfa, Biskra, S. Algeria), and
Lambessa siniscalchii TURATI, 1926, Atti Soc. ital. Sci. nat. 65: 29, figured (Locus typicus: Cirene; Derna
(Cirenaica) [Libya]). No material of these taxa could be examined and, therefore, they are tentatively left in
synonymy with Lasiocampa decolorata, following De FREINA & WITT 1987. The status of these taxa will be
clarified, when material from their type localities becomes available for molecular analysis. The taxon Lambessa
decolorata ssp. rubrescens WILTSHIRE, 1986, Fauna of Saudi Arabia 8: 268. Locus typicus: Wadi Sarawin, 610
m (Saudi Arabia) is now viewed as a subspecies of the Asian Lasiocampa puengeleri STERTZ, 1915
(LEWANDOWSKI & FISCHER 2012). A few specimens recorded as L. decolorata from Um Arad (Sinai) and a
few erroneous records from ‘Palestine’ (cf. LEWANDOWSKI & FISCHER 2012) are probably also referable to L.
puengeleri, the specimens from Sinai were stated to be “redder than KLUG’s type” (WILTSHIRE 1948, 228).
FIGURE 5. Lasiocampa decolorata (KLUG, 1830) ♀ Holotype, "Decolorata, N, Tscheila Ehrenb, 9018, BC ZSM Lep 81612.
Museum für Naturkunde Berlin"
FIGURE 6. Lasiocampa decolorata (KLUG, 1830) ♂ Syntype, "Lambessa decolorata sordidior Type Rothsch., syntype;
Guelt-es-Stel, C. Algeria, September 1913 (V. Faroult), Rothschild Bequest 1939-1". Natural History Museum London.
FIGURE 7. Lasiocampa decolorata (KLUG, 1830) ♀ Syntype. "Syntype, Guelt-es-Stel, Centr. Algeria (V. Faroult), 24. 9. 13,
Rothschild Bequest B. M. 1939-1". Natural History Museum London.
FIGURE 8. Lasiocampa hannae sp. n. ♂ Holotype. "Morocco, Anti Atlas Mts, Sidi M’zal village, 1260 m, 29°48‘00’’ N;
8°52’11’’ W, 30. ix. 2005, leg. Gy. M. László & G. Ronkay". MWM.
FIGURE 9. Lasiocampa editae sp. n. ♂ Holotype. "Morocco, High Atlas, Oukaimeden, NW slopes, 2,0–2,300 m, 11.–20. 08.
2012, G. Muller, E. Revay et al., BC ZSM Lep 76653, Genitalpräparat Heterocera Nr. 19872 Museum Witt München".
FIGURE 10. Lasiocampa editae sp. n. ♂ "Morocco, High Atlas, Oukaimeden, NW slopes, 2,0–2,300 m, 1.–10. 08. 2012, G.
Muller, E. Revay et al. " MWM.
FIGURE 11. Lasiocampa staudingeri (BETHUNE-BAKER, 1885) ♂ "Algerien, Juli, coll. Th. Witt, München/Weiden".
FIGURE 12. Lasiocampa staudingeri (BETHUNE-BAKER, 1885) ♀ "e. l. 11. x. 71, Tunisia, vic. Hammamet, G. Hesselbarth
leg., coll. Th. Witt, München/Weiden, Abgebildet de Freina & Witt, Bombyces & Sphinges der Westpalaearktis, Bd. 1, Taf. 28,
Fig. 41".
FIGURE 13. Lasiocampa staudingeri (BETHUNE-BAKER, 1885) ♀ "Algerien, Juli, coll. Th. Witt, München/Weiden".
FIGURE 14. Lasiocampa decolorata (KLUG, 1830) ♂ "Algier, Guelt es Stel, Franz Daniel". MWM.
FIGURE 15. Lasiocampa decolorata (KLUG, 1830) ♂ "Lasio. decolorata ♂ nov. var. Guelt es Stel, Museum Witt, München"
FIGURE 16. Lasiocampa decolorata (KLUG, 1830) ♂ "Algier, Guelt es Stel, Franz Daniel, Abgebildet de Freina & Witt,
Bombyces & Sphinges der Westpalaearktis Bd. 1, Taf. 28, Fig. 42". MWM.
FIGURE 17. Lasiocampa hannae sp. n. ♂ "Morocco, Anti Atlas Mts, Tizi n’ Tarakatine pass, 1600 m, 29°45’11’’N;
8°50’02’’W, 29. ix. 2005, leg. Gy. M. László & G. Ronkay, Museum Witt, München".
FIGURE 18. Lasiocampa hannae sp. n. ♂ "Morocco, Anti Atlas Mts, Sidi M’zal village, 1260 m, 29°48‘00’’ N; 8°52’11’’ W,
30. ix. 2005, leg. Gy. M. László & G. Ronkay, Museum Witt, München".
FIGURE 19. Lasiocampa hannae sp. n. ♂ "Morocco, Anti Atlas Mts, Sidi M’zal village, 1260 m, 29°48‘00’’ N; 8°52’11’’ W,
30. ix. 2005, leg. Gy. M. László & G. Ronkay, Museum Witt, München".
FIGURE 20. Lasiocampa editae sp. n. ♂ "Morocco, High Atlas, Oukaimeden NW slopes, 2,3–2,500 m, 01.–10.08.2012, G.
Muller, E. Revay et al. ". MWM.
FIGURE 21. Lasiocampa editae sp. n. ♂ "Morocco, High Atlas, Oukaimeden, NW slopes, 2,0–2,300 m, 11.–20. 08. 2012, G.
Muller, E. Revay et al.". MWM.
FIGURE 22. Lasiocampa editae sp. n. ♂ "Morocco, High Atlas, Oukaimeden NW slopes, 2,3–2,500 m, 01.–10.08.2012, G.
Muller, E. Revay et al. ". MWM.
FIGURE 23. Lasiocampa staudingeri (BETHUNE-BAKER, 1885) ♂ "Algerien, Lambesse, Korb, Algerien, Lambessa, Franz
Daniel, Abgebildet de Freina & Witt, Bombyces & Sphinges der Westpalaearktis Bd. 1, Taf. 28, Fig. 40." MWM"
FIGURE 24. Lasiocampa staudingeri (BETHUNE-BAKER, 1885) ♂ "Tunis, 11. A. Faller, Fbg, coll. Th. Witt, München/
Weiden, Abgebildet de Freina & Witt, Bombyces & Sphinges der Westpalaearktis Bd. 1, Taf. 28, Fig. 39, staudingeri Bak. ".
MWM.
FIGURE 25. Lasiocampa staudingeri (BETHUNE-BAKER, 1885) ♂ "Coll. A. Seitz, Algerien, Lambessa, coll. Seitz,
Genitalpräparat Heterocera Nr. 8691, Museum Wit München.
Zootaxa 3999 (3) © 2015 Magnolia Press · 407LASIOCAMPA DECOLORATA
Other records of L. decolorata from Egypt are also thought to be erroneous (WILTSHIRE 1948, 228). One male in
bad condition was doubtfully recorded as “L. decolorata” by RUNGS 1981 (page 428) from eastern Morocco,
Kenitra (elevation 25 m). It would be the only record for Morocco and the most western one, if verified.
Lambessa virago ROTHSCHILD, 1912 from Biskra, Col de Sfa (Algeria) may represent a specimen of
Lasiocampa decolorata modified by the influence of breeding abroad. It also seems likely that the taxon
siniscalchii is correctly placed as a synonym judging from its geographical origin between Algeria and Egypt.
Genetic data. BIN: BOLD: ABW5023 (n=3, 658bp), including the Egyptian holotype and two specimens
from Tunisia (identified as “subsp. datini”), intraspecific variation: 0.6%, suggesting conspecificity of the
populations from Egypt and Tunisia. In addition, one very short (94bp) DNA sequence from an Algerian specimen
(Guelt es Stel, “subsp. sordidior”), at 0.6% distance from the other sequences of BIN BOLD:ABW5023,
supporting the hypothesis of conspecificity. Closest neighbouring species: Lasiocampa editae with a genetic
distance of 3.5%, Lasiocampa hannae with 3.0% divergence. The (short 164bp) barcode-fragment of a wingless
female of L. staudingeri (Tunisia) only shows a divergence of 1.2%, but this value may change when the whole
barcode fragment of that species will be sequenced.
Lasiocampa hannae SPEIDEL, MOOSER & WITT sp. n.
(Figs 8, 17–19)
Material examined. Holotype ♂: Morocco, Anti Atlas Mts, Sidi M’zal village, 1260 m, 29°48‘00’’ N; 8°52’11’’
W, 30. ix. 2005, leg. Gy. M. László & G. Ronkay, Museum Witt, München. BC ZSM Lep 76624, genitalia slide
Heterocera Nr. 19873 Museum Witt München
Paratypes: 2 ♂ dto, one with additional labels, DNA-6456-2010-73. coll. MWM, 2 ♂ dto. coll. Mooser, 1 ♂
Morocco, Anti Atlas Mts, Tizi n’ Tarakatine pass, 1600 m, 29°45’11’’N; 8°50’02’’W, 29. ix. 2005, leg. Gy. M.
László & G. Ronkay, Museum Witt, München.
Description. Wingspan 36–40 mm, forewing length 16–19 mm. Wing-shape and structure of antenna
corresponding to that of L. trifolii. Easily distinguished from the pale taxa of the L. decolorata complex by its
reddish-brown wing color, very similar to L. editae sp. n. but with a smaller average wingspan and a slightly paler
color.
Male genitalia (genitalia slide 19873 (MWM), fig. 2): Valvae very modified, like in all Lasiocampa species in
a hanging position, not laterally movable. The genitalia are easily damaged if one tries to move the valvae to a
standardized position and if the three–dimensional (steric) structures are too strongly flattened. Valva bifid. The
exterior lobe of the valva variable in size, strongly sclerotized. Interior lobe much stronger than in L. editae,
terminally ending truncate, broader. Vesica dorsolaterally everted, with the sclerotized zone stronger developed
than in L. editae.
Genetic data. BIN: ABX3741 (n=2), including holotype. Intraspecific variation: 0.0%. Closest neighbouring
species: Lasiocampa decolorata at a genetic distance of 3.0%, Lasiocampa hannae at 3.2%.
Etymology. Named in honor of Mrs. Hanna Sander-Mooser.
Ecology. Flight period September, at medium elevations of 1200–1600 m a.s.l. in an isolated area of the Anti
Atlas Mountains.
Lasiocampa editae SPEIDEL, MOOSER & WITT sp. n.
(Figs 9, 20–22)
Material. Holotype ♂: Morocco, High Atlas, Oukaimeden, NW slopes, 2,0–2,300 m, 11.–20. 08. 2012, G. Muller,
E. Revay et al., BC ZSM Lep 76653, Genitalpräparat Heterocera Nr. 19872 Museum Witt München.
Paratypes: 10 ♂ dto., with additional labels Genitalpräparat Heterocera Nr. 19869 Museum Witt München and
BC ZSM Lep 76656; Genitalpräparat Heterocera Nr. 19871 Museum Witt München and BC ZSM Lep 76654;
Genitalpräparat Heterocera Nr. 19868 Museum Witt München and BC ZSM Lep 76651; Genitalpräparat
Heterocera Nr. 19870 Museum Witt München and BC ZSM 76655; Genitalpräparat Heterocera Nr. 19867 Museum
Witt München and BC ZSM Lep 76652. Museum Witt München; dto. 6 ♂ 01.–10. 08. 2013. Museum Witt
München; 2 ♂ Morocco, High Atlas, Oukaimeden, Tisrafene, 2,8–3000 m, mid Aug. 2013, G. Muller, E. Revay et
SPEIDEL ET AL. 408 · Zootaxa 3999 (3) © 2015 Magnolia Press
al. Museum Witt München; 10 ♂ Morocco Toubkal, Ait El Qad 2200–2600 m, 22–31. 7. 2012, G. Muller, E Revay
et al. Coll. Mooser; 3 ♂ Morocco Toubkal, Ait El Qad 2200–2600 m, 10–15. 7. 2012, G. Muller, E Revay et al.
Coll. Mooser; 2 ♂ Morocco, High Atlas, Oukaimeden, southern slopes, 2300–2500 m, 5–10. 7. 2012, G. Muller, E
Revay et al. Coll. Mooser; 19 ♂ Morocco, High Atlas, Oukaimeden, southern slopes, 2300–2500 m, 22–31. 7.
2012, G. Muller, E Revay et al. Coll. Mooser; 4 ♂ Morocco, High Atlas, Oukaimeden NW slopes, 2,0–2,300 m,
11.–20.08.2012, G. Muller, E. Revay et al. MWM; 10 ♂ Morocco, High Atlas, Oukaimeden NW slopes, 2,3–2,500
m, 1.–10.08.2012, G. Muller, E. Revay et al. MWM.
Description. Male wingspan 38–46 mm, forewing length 19–22 mm. Wing-shape and antenna-structure
corresponding with that of other members of the L. trifolii group, but without traces of fasciae. Wings reddish
brown, superficially indiscriminable from L. hannae sp. n., but on average slightly larger and darker than the latter.
Male genitalia (genitalia slides 19867-19872 (MWM), fig. 3): Valvae very modified, like in all Lasiocampa
species in a dorsal position in the standard preparations, not laterally movable. The genitalia are easily damaged, if
one tries to move the valvae to a standardized position and if the three–dimensional structures are too strongly
flattened. Valva bifid. The exterior lobe of the valva is variable in size, strongly sclerotized. Interior lobe weaker,
rounded and easily destroyed during preparation. Vesica dorsolaterally everted.
Genetic data. BIN: ACL1902 (n=6), including holotype. Intraspecific variation: 0.0%. Closest neighbouring
species: Lasiocampa decolorata at a genetic distance of 3.5%, Lasiocampa hannae at 3.2%.
Etymology. Named in honour of Prof. Dr. Dr. Edita Revay.
Ecology. The species is found in the Central High Atlas Mts. above the tree line, adults fly in July and August.
For habitat pictures see Figs 26 and 27.
Discussion. The females of the two new species are so far unknown. Females of the Lasiocampa trifolii group
are readily attracted to light, and Lasiocampa decolorata also possesses winged females that are phototrophic.
Thus, the females of the present species may show low affinity to light or even may prove to be wingless, like the
allied species Lasiocampa staudingeri which ranges from Libya to Morocco. However, males of the latter taxon are
easily discriminated by its paler yellow rather than reddish-brown wing colour. As well, adults are only found in
autumn and there are no records from the alpine region.
Lasiocampa staudingeri (BETHUNE-BAKER, 1885)
(Figs 11, 23–25)
Bombyx staudingeri BETHUNE-BAKER, 1885, Entomologist's mon. Mag. 21: 242. Locus typicus: Lambessa (Algeria)
[author name only Baker in the original publication]
Synonym:
Lambessa staudingeri nom. nov. castaneolavata ROTHSCHILD, 1914, Novit. zool. 21: 314. Locus typicus: Hauts Plateaux,
Prov. Oran (Sebdou).
Material. 2 ♂ Algeria, Lambessa. MWM, 1 ♂ Algeria (no further data); 1 ♂ Tunis, 11. A. Faller, Fbg, coll. Th.
Witt, München/Weiden, Abgebildet de Freina & Witt, Bombyces & Sphinges der Westpalaearktis Bd. 1, Taf. 28,
Fig. 39, staudingeri Bak. MWM; 1 ♀ Tunis e. l., [Kurt John], Franz Daniel; 1 ♀ e. l. 11. x. 71, Tunisia, vic.
Hammamet, G. Hesselbarth leg., Sammlung de Freina München, coll. Th. Witt München/Weiden, Abgebildet de
Freina & Witt, Bombyces & Sphinges der Westpalaearktis Bd. 1, Taf. 28, Fig. 41, BC ZSM Lep 84742.
Male: The male adults are similar to the other species described here, although the ground colour is
distinctively paler than in the other species of the L. staudingeri-group, unicolourous pale yellowish brown. Female
with only minute traces of wings, almost wingless.
Male genitalia (genitalia slide 8691 (MWM), fig. 4). The aedeagus is slightly curved, whereas it is straight in
the other species. Vesica more or less terminally everted. It is likely that cross-breeding with L. decolorata and
allied species is improbable due to different form of the aedeagus and orientation of the everted vesica.
Distribution. The species is known from Libya to Morocco (De FREINA & WITT 1987), but many of the
localities require confirmation, as males can be confused with Lasiocampa decolorata.
Genetic data: A 164bp COI-fragment was obtained from a wingless female from northern Tunisia. It showed
1.2% divergence from Tunisian/Algerian Lasiocampa decolorata, 2.5% from L. hannae sp. n., and 3.7% from L.
editae sp. n., supporting species status of the last two.
Zootaxa 3999 (3) © 2015 Magnolia Press · 409LASIOCAMPA DECOLORATA
FIGURE 26. Morocco, Toubkal, Ait El Qad 2200–2600 m. Typical habitat of L. editae on the southern slopes of the High Atlas
Mts. Here Dr. Revay collecting Coenonympha vaucheri, Berberia abdelkader and Hipparchia hansii, typical butterfly species
associated with L. editae.
FIGURE 27. Morocco, High Atlas, Oukaimeden, Tisrafene south facing slopes, 2,800–3,000m.
SPEIDEL ET AL. 410 · Zootaxa 3999 (3) © 2015 Magnolia Press
FIGURE 28. NJT tree (Kimura 2 parameter, built with MEGA6) for 10 species in the genus Lasiocampa based on sequence
variation at COI. Sequence length of the holotype of Lasiocampa decolorata: 658bp. Width of triangles represent sample size,
depth the genetic variation within the cluster. Source: DNA Barcode data from BOLD (Barcode of Life Database, cf.
RATNASINGHAM & HEBERT 2007).
Discussion. The present species is surprisingly similar to Lasiocampa decolorata in its COI fragment. It seems
questionable whether L. staudingeri is a separate species, when only considering the small genetic divergence of
1,2% which concerns, however, just the short length of the 164bp fragment. There may be more informative
nucleotide sites in the rest of the barcode region. Moreover, Lasiocampa decolorata has full-winged females
whereas those of L. staudingeri have strongly reduced wings. Finally there are subtle morphological differences as
given above. We conclude that there are good arguments for tentatively retaining the two taxa as different species.
More data are needed for a definitive decision about the status of L. staudingeri as a different biospecies or not, but
in any case the putative close relationship of species/populations with winged and almost wingless females is
surprising.
Acknowledgements
We are grateful to the Ministry of Agriculture, Morocco for permission to carry out this work and to the General
Secretariat of Forestry and the local authorities for their field support. We want to thank Dr. Wolfram Mey,
Museum für Naturkunde, for the loan of the type specimen of Lasiocampa decolorata. Mr. Tibor Csövari kindly
and carefully prepared many of the genitalia slides and the photos of them, whereas the remaining genitalia slides
were made by Mrs. Mei-Yu Chen and photographed by our colleague Mr. Ulf Buchsbaum. Mr. Harald Sulak and
Mrs. Mei-Yu Chen prepared the colour figures, Mr. Igor Kostyuk assembled the plate.
Literature
Baker, D.B. (1997) C.G. Ehrenberg and W.F. Hemprich’s travels, 1820–1825, and the Insecta of the Symbolae Physicae.
Deutsche Entomologische Zeitschrift, 44, 165–202.
http://dx.doi.org/10.1002/mmnd.19970440208
Zootaxa 3999 (3) © 2015 Magnolia Press · 411LASIOCAMPA DECOLORATA
Baker, G.T. (1885) Descriptions of some new species of Lepidoptera from Algeria. Entomologist's Monthly Magazine, 21, 268–
270.
Dewaard, J.R., Ivanova, N.V., Hajibabaei, M. & Hebert, P.D.N. (2008) Assembling DNA barcodes: analytical protocols. In:
Martin, C. (Ed.), Methods in molecular biology: environmental Genetic data. Humana Press, Totowa, New Jersey, pp.
275–293.
http://dx.doi.org/10.1007/978-1-59745-548-0_15
Ehrenberg, C.G. (1828) Naturgeschichtliche Reisen durch Nord-Afrika und West-Asien in den Jahren 1820 bis 1825 von Dr. W.
F. Hemprich und Dr. C.G. Ehrenberg. Historischer Teil. Reisen in Aegypten, Libyen, Nubien und Dongala 1 (1). Ernst
Siegfried Mittler, Berlin, Posen und Bromberg, xxx + 162 pp.
Freina, J. de & Witt, T.J. (1987) Die Bombyces und Sphinges der Westpalaearktis. Edition Forschung und Wissenschaft Verlag,
München, 708 pp.
Goldstein, P.Z. & deSalle, R. (2011) Integrating DNA barcode data and taxonomic practice: Determination, discovery, and
description. Bioessays, 33, 135–147.
http://dx.doi.org/10.1002/bies.201000036
Hausmann, A., Hebert, P., Mitchell, A., Rougerie, R., Sommerer, M., Edwards, T. & Young, C.J. (2009a) Revision of the
Australian Oenochroma vinaria Guenée, 1858 species-complex (Lepidoptera, Geometridae, Oenochrominae): DNA
barcoding reveals cryptic diversity and assesses status of type specimen without dissection. Zootaxa, 2239, 1–21.
Hausmann, A., Sommerer, M., Rougerie, R. & Hebert, P. (2009b) Hypobapta tachyhalotaria n. sp. from Tasmania – an
example of a new species revealed by DNA barcoding (Lepidoptera, Geometridae). Spixiana, 32 (2), 237–242.
Hausmann, A. (2011) An integrative taxonomic approach to resolving some difficult questions in the Larentiinae of the
Mediterranean region (Lepidoptera, Geometridae). Mitteilungen der Münchner Entomologischen Gesellschaft, 101, 73–
97.
Hebert, P.D.N., Ratnasingham, S. & DeWaard, J.R. (2003) Barcoding animal life: cytochrome c oxidase subunit 1 divergences
among closely related species. Proceedings of the Royal Society of London, Series B: Biological Sciences, 270
(Supplement 1), 96–99.
http://dx.doi.org/10.1098/rsbl.2003.0025
Hebert, P.D.N., Penton, E.H., Burns, J.M., Janzen, D.H. & Hallwachs, W. (2004) Ten species in one: DNA barcoding reveals
cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proceedings of the National Academy of Sciences,
101, 14812–14817. [USA]
http://dx.doi.org/10.1073/pnas.0406166101
Ivanova, N.V., DeWaard, J.R. & Hebert, P.D.N. (2006) An inexpensive, automation-friendly protocol for recovering high-
quality DNA. Molecular Ecology Notes, 6, 998–1002.
http://dx.doi.org/10.1111/j.1471-8286.2006.01428.x
Jörger, K.M. & Schrödl, M. (2014) Opinion – DNA taxonomy '2.0'. Spixiana, 37 (1), 20.
Klug, F. (1830) Symbolae physicae seu Icones et Descriptiones Insectorum [quae] ex itinere [per Africam borealem et Asiam
occidentalem Friderici Guilelmi] Hemprich et [Christiani Godofredi] Ehrenberg [... studio novae aut illustratae
redierunt] / percensuit Dr. Fr. Klug. – Decas 2. [ex officina academica. Venditur a] Mittlero, Berolini, plates 14–20.
[unpaginated]
Lewandowski, S. & Fischer, H. (2005) Revision der Artengruppen von Lasiocampa trifolii und L. serrula der Gattung
Lasiocampa von Paula Schrank, 1802 (Lepidoptera: Lasiocampidae). Nachrichten des Entomologischen Vereins Apollo,
New Series, 26, 183–196.
Lewandowski, S. & Fischer, H. (2008) Nachträge zur Revision der Artengruppen von Lasiocampa trifolii und L. serrula der
Gattung Lasiocampa von Paula Schrank, 1802 (Lepidoptera: Lasiocampidae). Nachrichten des Entomologischen Vereins
Apollo, New Series, 29, 33–41.
Lewandowski, S. & Fischer, H. (2012) Zweiter Nachtrag zu den Revisionen der trifolii- und serrula-Artengruppen der Gattung
Lasiocampa sowie neue Erkenntnisse zum Typenfundort von L. decolorata (Lepidoptera: Lasiocampidae). Nachrichten
des Entomologischen Vereins Apollo, New Series, 32, 97–104.
Mabille, M.P. (1888) Description de quelques Lépidopoptères nouveaux de Tunisie et d’Algérie. Annales de la Société
Entomologique de France, (6) 8 (Bull.): xlii–xliii.
Oberthür, C. (1890) Lépidoptères d’Algérie. Études d’Entomologie 13, 19–33.
Prosser, S.W.J., DeWaard, J.R., Miller, S.E. & Hebert, P.D.N. (2015) DNA barcodes from century-old type specimens using
next generation sequencing. PLoS ONE. [in review]
Padial, J.M., Miralles, A., De La Riva, I. & Vences, M. (2010) The integrative future of taxonomy. Frontiers in Zoology, 7, 16.
http://dx.doi.org/10.1186/1742-9994-7-16
Ratnasingham, S. & Hebert, P.D.N. (2007) BOLD: The Barcode of Life Data System (http://www.barcodinglife.org).
Molecular Ecology Notes, 7 (3), 355–364.
http://dx.doi.org/10.1111/j.1471-8286.2007.01678.x
Robinson, G.S. (1976) The preparation of slides of Lepidoptera genitalia with special reference to the Microlepidoptera.
Entomologist’s Gazette, 27, 127–132.
Rothschild, W. (1912) On an apparently new form of Lambessa from Algeria. Novititates Zoologicae, 19, 118.
Rothschild, W. (1914) A preliminary account of the Lepidopterous fauna of Guelt-Es-Stel, central Algeria. Novitates
SPEIDEL ET AL. 412 · Zootaxa 3999 (3) © 2015 Magnolia Press
Zoologicae, 21, 299–357.
Rungs, C.E.E. (1981) Catalogue Raisonné des Lepidoptères du Maroc. 2. Travaux de l’ Institut Scientifique, Série Zoology, 40,
223–588. [Rabat]
Jörger, K.M. & Schrödl, M. (2014) Opinion. Taxonomy ‘2.0’. Spixiana, 37 (1), 20.
Tautz, D., Arctander, P., Minelli, A., Thomas, R.H. & Vogler, A.P. (2002) DNA points the way ahead in taxonomy. Nature, 418,
479.
http://dx.doi.org/10.1038/418479a
Tautz, D., Arctander, P., Minelli, A., Thomas, R.H. & Vogler, A.P. (2003) A plea for DNA taxonomy. Trends in Ecology &
Evolution, 18 (2), 70–74.
http://dx.doi.org/10.1016/S0169-5347(02)00041-1
Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013) MEGA6: Molecular Evolutionary Genetics Analysis
Version 6.0. Molecular Biology and Evolution, 30, 2725–2729.
http://dx.doi.org/10.1093/molbev/mst197
Teletchea, F. (2010) After 7 years and 1000 citations: Comparative assessment of the DNA barcoding and the DNA taxonomy
proposals for taxonomists and non-taxonomists. Mitochondrial DNA, 21 (6), 206–226.
http://dx.doi.org/10.3109/19401736.2010.532212
Turati, E. (1926) Novita di Lepidopterologia in Cirenaica. Atti della Societa Italiana di Scienze Naturali e del Museo Civico di
Storia Naturale in Milano, 65, 25–83.
Will, K.W., Mishler, B.D. & Wheeler, Q.D. (2005) The perils of DNA barcoding and the need for integrative taxonomy.
Systematic Biology, 54 (5), 844–51.
http://dx.doi.org/10.1080/10635150500354878
Wiltshire, E.P. (1948) The Lepidoptera of the Kingdom of Egypt. Part I. Bulletin de la Société Fouad Ier d’Entomologie, 32,
203–296, 7 pls.
Wiltshire, E.P. (1986) Lepidoptera of Saudi Arabia. Fam. Cossidae, Sesiidae, Metarbelidae, Lasiocampidae, Sphingidae,
Geometridae, Lymantriidae, Arctiidae, Nolidae, Noctuidae (Heterocera); Fam. Satyridae (Rhopalocera) (Pt. 5). Fauna of
Saudi Arabia 8, 262–323.
