2urn:lsid:arphahub.com:pub:45048D35-BB1D-5CE8-9668-537E44BD4C7Eurn:lsid:zoobank.org:pub:91BD42D4-90F1-4B45-9350-EEF175B1727AZooKeysZK1313-29891313-2970Pensoft Publishers10.3897/zookeys.843.3286732867Research ArticleAnimaliaGastropodaHelicidaeInvertebrataMolluscaStylommatophoraNomenclatureSystematicsTaxonomyAfricaEuropeAnatomical and phylogenetic investigation of the genera Alabastrina Kobelt, 1904, Siretia Pallary, 1926, and Otala Schumacher, 1817 (Stylommatophora, Helicidae)KneubühlerJeannettejeannette.kneubuehler@students.unibe.ch12HuttererRainer3PfarrerBeat12NeubertEikehttps://orcid.org/0000-0002-0277-289412Naturhistorisches Museum der Burgergemeinde Bern, 3005 Bern, SwitzerlandNaturhistorisches Museum der Burgergemeinde BernBernSwitzerlandInstitute of Ecology and Evolution, University of Bern, 3012 Bern, SwitzerlandUniversity of BernBernSwitzerlandZoologisches Forschungsmuseum Alexander Koenig, 53113 Bonn, GermanyZoologisches Forschungsmuseum Alexander KoenigBonnGermany
2019090520198431379CBEDB07-7FD6-5464-8608-FC12B0D9F1C04821C74E-C7E8-4866-9BAE-9A03CC46AD1634849520401201901042019Jeannette Kneubühler, Rainer Hutterer, Beat Pfarrer, Eike NeubertThis 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.http://zoobank.org/4821C74E-C7E8-4866-9BAE-9A03CC46AD16
This study presents new insights in the anatomy of genital organs of some large helicid gastropods from northern Africa. The genetic analysis with the markers COI, 16S, H3, and 5.8 S rRNA+ITS2 reveales a high support for Alabastrina and Otala as separate evolutionary lineages within the Otalini. The position of Siretia as another separate lineage within the Otalini is discussed. “Tingitanaminetteidecussata” clusters within the O.xanthodon clade and confirms that the genus Tingitana can be synonymised with Otala. The genus Alabastrina differs from all other known genera by possession of a penial appendix. This character state is also found in topotypic A.tistutensis. Examination of the twin penial papilla system in Otala recovers a reduction of the proximal penial papilla in O.punctata. The position of Helixmurcica as a separate subspecies of O.lactea is not supported, and it is here considered to be a synonym of the latter species.
Kneubühler J, Hutterer R, Pfarrer B, Neubert E (2019) Anatomical and phylogenetic investigation of the genera Alabastrina Kobelt, 1904, Siretia Pallary, 1926, and Otala Schumacher, 1817 (Stylommatophora, Helicidae). ZooKeys 843: 1–37. https://doi.org/10.3897/zookeys.843.32867
Introduction
Working with the terrestrial molluscs from northern Africa, students are faced with a confusing situation: an enormous number of species- and genus-level taxa are available to arrange the malacodiversity but for many groups a modern treatment is missing. As a result, this important part of the Palaearctic fauna is still in a chaotic state (Rour et al. 2002). The major problem in the Helicidae is the absence of a stable generic concept that is based on recognisable character states. This can be morphological, anatomical, or genetic data. For this reason, we follow the idea of integrative taxonomy and try to draw conclusions based on a synopsis of these types of traits.
Research on the malacofauna of northern Africa was mainly elaborated by three researchers, Bourguignat (1829–1892), Kobelt (1840–1916), and Pallary (1869–1942), who laid a fundament so strict that it is followed more or less until today. This system was more or less supported by P Hesse (1911) by his anatomical research on some groups of Helicidae. His research was the onset of the valorisation of genital morphology as another source of characters and character states. Amongst others, he investigated species, which are treated also in this publication. Unfortunately, Hesse restricted his research to the outer morphology of the genital organs thus missing the highly informative traits found in the lumen. While in the remaining part of the western Palaearctic, taxonomy of terrestrial snails went through a phase of deep changes, northern Africa was left more or less untouched. This situation is currently changing, and several papers were published in the last years which resulted in new data, for example on the Helicidae (Psonis et al. 2013, Neubert 2014, Neubert and Korábek 2015, Walther et al. 2016, Bouaziz-Yahiatene et al. 2017). Recently, Holyoak and Holyoak (2017) published a major paper on the large group of Otalini G Pfeffer, 1930, which has its centre of radiation in the north-west of Africa. In this paper, the authors went through numerous available names and came up with a radical solution following a lumping approach.
The investigation in this study is mainly based on specimens collected by the second author during his excavation campaigns in north-eastern Morocco (Hutterer et al. 2011a, b. 2014). The taxonomic investigation of terrestrial molluscs was part of an archaeological study of various cave sediments in the Rif region (Mikdad et al. 2000).
This study aims to serve as an addition to the recent studies on helicid phylogeny. Due to the restricted number of taxa available in our study, we here can add only some remarks to the ongoing work on the north African Helicidae. Particular emphasis is laid on filling gaps in the knowledge of the anatomy of the genital organs. It has to be stressed that the investigation of this complex of organs should always include the structure of the internal lumina; they certainly help in identifying autapomorphic character states. In addition, we supply new data on shell and anatomical traits, and present a first genetic approach to some of the genera involved using the following markers: cytochrome c oxidase subunit I (COI), 16S rRNA (16S), histone 3 (H3), and partial sequence of 5.8 S rRNA flanking the internal transcribed spacer 2 (ITS2).
Material and methodsSpecimens investigated
The specimens were collected in Morocco and Algeria between 1998 and 2015. Reference specimens from Spain and Portugal could be included. Detailed sampling locations of the investigated specimens are given in Fig. 1 and Table 1. The voucher number and the GenBank accession numbers for the obtained DNA sequences can be found in Table 1. All specimens used in this study are housed in the Natural History Museum Bern, Switzerland.
Sampling locations of the investigated specimens. This map was produced with QGIS (2016, v2.18.12) using the Natural Earth data set.
https://binary.pensoft.net/fig/299955
Detailed list of the sampling sites and the GenBank accession numbers of the investigated specimens.
Species
Locality
Latitude
Longitude
Voucher
GenBank accession number COI
GenBank accession number 16S
GenBank accession number H3
GenBank accession number ITS2
Alabastrinaalabastrites
Morocco, Montes de Kebdana, Kebdana Mountain/ Rif
35.027N, 2.614W
NMBE-549817
MK754458
MK585087
MK728781
MK585111
Morocco, Rif Jbel Fiztoutine w Hills El Batel
34.938N, 3.193W
NMBE-549813
MK754457
MK585086
MK728780
MK585110
Morocco, Cave Ifri n’Ammar, 20 km SW Berkane
34.782N, 3.094W
NMBE-549812
MK754456
MK585085
MK728779
MK585109
Morocco, Hassi Ouenzga nach Afso/ Oriental
34.796N, 3.195W
NMBE-549811
MK754455
MK585084
MK728778
MK585108
Morocco, Etsedda/ Kebdane
35.195N, 3.269W
NMBE-549816
MK754459
MK585088
MK728782
MK585112
Alabastrinatistutensis
Morocco, Rif, Tiztoutine, village bouaza
34.955N, 3.166W
NMBE-555174
MK754469
MK585099
MK728792
MK585123
Allognathusbalearicus
Spain, Mallorca, Escorça
39.822N, 2.887E
EHUMC-1051
KR705026
KR704986
no data
no data
Ariantaarbustorum
Austria, Upper Austria, Höllengebirge Mts
no data
no data
NHM-109000
KF596871
KF596823
KF596915
no data
Helixmelanostoma
Tunisia, Kasserine
35.172N, 8.831E
NMBE-540550
MF564162
MF564116
MF564178
no data
France, between Rabieux and Saint-Félix-de-Lodez/ Herault
43.663N, 3.441E
NMBE-520822
MK754471
MF564115
MF564177
no data
Marmoranamuralis
Italy, Rome
41.885N, 12.481E
MN-2554
KR705023
KR704983
no data
no data
Massylaeaconstantina
Algeria, Ighil Bourmi
36.487N, 4.061E
NMBE-540545
MF564168
MF564122
MF564185
no data
Massylaeavermiculata
Algeria, Makouda, Tizi Ouzou/ Kabylie
36.791N, 4.066E
NMBE-540544
MF564159
MF564112
MF564174
no data
Otalalactea
Spain, Finca de la Concepción, N Málaga
36.760N, 4.428W
NMBE-554174
MK754463
MK585093
MK728786
MK585117
Spain, Punta Entinas, W Almería
36.690N, 2.694W
NMBE-554175
MK754464
MK585094
MK728787
MK585118
Spain, Punta Entinas, W Almería
36.690N, 2.694W
NMBE-554176
MK754465
MK585095
MK728788
MK585119
Portugal, W Almocageme/ Sintra Cascais National Park
38.798N, 9.485W
NMBE-553246
MK754460
MK585089
MK728783
MK585113
Morocco, Hassi Ouenzga/ Oriental
34.698N, 3.256W
NMBE-555171
MK754452
MK585081
MK728775
MK585105
Morocco, Hassi Ouenzga/ Oriental
34.698N, 3.256W
NMBE-549814
MK754468
MK585098
MK728791
MK585122
Morocco, West of Aoulouz/ Souss-Massa-Draa
30.709N, 8.268W
NMBE-549951
MK754472
MK603015
MK728794
MK602877
Morocco, Etsedda/ Kebdane
35.195N, 3.269W
NMBE-545594
MK754448
MK585077
MK728771
MK585101
Otalapunctata
Spain, El Tarajal, W Málaga
36.705N, 4.506W
NMBE-554171
MK754462
MK585092
MK728785
MK585116
Spain, El Tarajal, W Málaga
36.705N, 4.506W
NMBE-554172
MK754467
MK585097
MK728790
MK585121
Algeria, Makouda, Tizi Ouzou/ Kabylie
36.745N, 4.068E
NMBE-534228
MK754466
MK585096
MK728789
MK585120
Otalatingitana
Morocco, Tarzout de Guigou/ Boulmane, NW Boulmane
33.381N, 4.778E
NMBE-510549
no data
no data
no data
no data
Otalaxanthodon
Morocco, Kebdana, Moulouya valley S Mechraa Elmalh
34.821N, 2.745W
NMBE-555169
MK754450
MK585079
MK728773
MK585103
Morocco, Kebdana, Moulouya valley S Mechraa Elmalh
34.821N, 2.745W
NMBE-555170
MK754451
MK585080
MK728774
MK585104
Kebdana, Moulouya valley below barrage
34.739N, 2.803W
NMBE-549825
MK754453
MK585082
MK728776
MK585106
Kebdana, Moulouya valley below barrage
34.739N, 2.803W
NMBE-549826
MK754454
MK585083
MK728777
MK585107
Morocco, Montes de Kebdana, Kebdana Mountain/ Rif
35.027N, 2.614W
NMBE-549841
MK754473
MK603016
MK728795
MK602878
Morocco, Montes de Kebdana, Djebel Sebaa Reyal/ Rif
35.030N, 2.613W
NMBE-549843
MK754474
MK603017
MK728796
MK602879
Morocco, Guercif, Oued Melloulon/ Taza al-Hoceima
34.207N, 3.414W
NMBE-549820
MK754449
MK585078
MK728772
MK585102
Siretiapallaryi
Morocco, Montes de Kebdana, Kebdana Mountain/ Rif
35.027N, 2.614W
NMBE-549815
MK754461
MK585090
MK728784
MK585114
Thebasubdentatasubdentata
Morocco, West of Aoulouz/ Souss-Massa-Draa
30.709N, 8.268W
NMBE-549949
MF564172
MF564126
MF564188
no data
“Tingitanaminetteidecussata”
Morocco, Montes de Kebdana, Djebel Sebaa Reyal/ Rif
35.030N, 2.613W
NMBE-549840
MK754470
MK585100
MK728793
MK585124
Abbreviations of institution:
MHNL Musée de Confluence, Lyon
MNHNMuseum National d’Histoire Naturelle, Paris
NMBENaturhistorisches Museum, Bern
SMF Research Institute Senckenberg, Frankfurt
Molecular study
For total DNA extraction the Qiagen Blood and Tissue Kit (Qiagen; Hilden, Germany) was used in combination with a QIAcube extraction robot. Ca. 0.5 cm3 of foot tissue was cut from the foot muscle and placed in a mix of 180 µl ATL buffer and 20 µl Proteinase K. It was then incubated for ca. 4 hours at 56 °C in a heater (Labnet, Vortemp 56, witec AG, Littau, Switzerland). For subsequent DNA extraction the QIAcube extraction robot with the Protocol 430 (DNeasy Blood Tissue and Rodent tails Standard) was used. In this study, two mitochondrial markers (COI and 16S) and two nuclear markers (H3 and 5.8 S rRNA+ITS2) were investigated. PCR mixtures consisted of 12.5 µl GoTaq G2 HotStart Green Master Mix (Promega M7423), 8.5 µl ddH2O, 1 µl forward and reverse primer each, and 2 µl DNA template. In Table 2 the used primer pairs for the PCR are listed. Following PCR cycles were used: for COI 2 min at 94 °C, followed by 35 cycles of 1 min at 95 °C, 1 min at 40 °C and 1 min at 72 °C and finally, 5 min at 72 °C; for 16S 5 min at 95 °C, followed by 45 cycles of 30 s at 95 °C, 30 s at 48 °C and 45 s at 72 °C, and finally, 5 min at 72 °C; for H3 3 min at 95 °C, followed 40 cycles of 1 min at 95 °C, 1 min at 42 °C and 1 min at 72 °C, and finally, 10 min at 72 °C, and for 5.8 S rRNA+ITS2 1 min at 96 °C, followed by 45 cycles of 30 s at 94 °C, 30 s at 50 °C and 1 min at 72 °C, and finally, 10 min at 72 °C (SensoQuest Tabcyclet and Techne TC-512, witec AG, Littau, Switzerland). The purification and sequencing of the PCR product was performed by LGC (LGC Genomics Berlin, Germany). Interpretation of Bootstrap values: 70 to 80 = moderate support; 80 to 90 = well supported; > 90 = high support. Bayesian posterior probabilities: values above 0.95 are significant support.
Used primer pairs for the two mitochondrial and two nuclear markers.
Gene
Primer
Sequence
Sequence length (bp)
Reference
COI
LCO1490
5’-GGTCAACAAATCATAAAGATATTGG-3’
680
Folmer et al. 1994
HCO2198
5’-TAAACTTCAGGGTGACCAAAAAATCA-3’
16S
16S ar
5’-CGC CTG TTT ATC AAA AAC AT-3’
440
Simon et al. 1994
16S br
5’- CCG GTC TGA ACT CTG ATC AT -3’
H3
H3AD
5’-ATGGCTCGTACCAAGCAGACVGC-3’
380
Colgan et al. 1998
H3BD
5’-ATATCCTTRGGCATRATRGTGAC-3’
ITS2
ITS2ModA
5’-GCTTGCGGAGAATTAATGTGAA-3’
900
Bouaziz-Yahiatene et al. 2017
ITS2ModB
5’-GGTACCTTGTTCGCTATCGGA-3’
Phylogenetic analyses
For the phylogenetic analyses sequences obtained from GenBank were included as outgroups: Ariantaarbustorum (Linnaeus, 1758) (Cadahia et al. 2014), Marmoranamuralis (OF Müller, 1774), and Allognathusbalearicus (Rossmässler, 1838) (= Allognathushispanicus (Rossmässler, 1838)) (Neiber and Hausdorf 2015). Additionally, sequences of Helixmelanostoma Draparnaud, 1801, Thebasubdentatasubdentata (Férussac, 1821), Massylaeaconstantina (E Forbes, 1838) and Massylaeavermiculata (OF Müller, 1774) from the study of Bouaziz-Yahiatene et al. 2017 were also included as outgroups. These species were selected to identify the phylogenetic placement of the focal taxa investigated in this study.
For sequence processing and editing the software package Geneious v9.1.8 (Biomatters Ltd) was used. The protein-coding gene fragments of COI and H3 were defined in two data blocks. The first two codon positions were defined as one block and the third codon position as a second block. The non-coding regions from 16S and 5.8 S rRNA+ITS2 were defined as a single data block. Partitionfinder-2.1.1 (Lanfear et al. 2012) was applied for searching optimal evolutionary models for the partitions using the corrected Akaike Information Criterion (cAIC). RAxML plug-in for Geneious (Stamatakis 2006) was implemented for computing ML inference, using Geneious’ plug-in with rapid bootstrapping setting, the search for the best scoring ML tree and 1500 bootstrapping replicates. Bayesian Inference (BI) was performed using Mr. Bayes v3.2.6 ×64 (Huelsenbeck and Ronquist 2001; Ronquist and Huelsenbeck 2003; Altekar et al. 2004) through the HPC cluster from the University of Bern (http://www.id.unibe.ch/hpc). For the concatenated data set, Partitionfinder-2.1.1 was used for finding the optimal evolutionary models for each subset with the model = all function. The Monte Carlo Markov Chain (MCMC) parameter was set as follows: starting with four chains and four separate runs for 20 million generations with a tree sampling frequency of 1000 and a burn in of 25%.
Anatomical and morphological study
Living animals were killed in boiling water and stored for one day in 80% ethanol. The next day, the ethanol was exchanged and the specimens were stored in the fridge at 5 °C until DNA extraction and dissection. Our experience showed that this procedure maintains the soft tissue and is essential for proper anatomical studies, as well as for the conservation of DNA. The dissection of the snail genitalia took place under a stereomicroscope (Leica MZ12) using thin tweezers and scissors. The genitalia were dissected from the body, spread on a wax bedded bowl, and properly pinned with small needles. The total length of the situs was measured using a calliper (Mitutoyo). Proportions between different parts of the genitalia were estimated using the total situs length as a reference. Additionally, the inner structures of the penis and the epiphallus were investigated. Pictures of the situs were taken with a Leica DFC425 microscope camera using an image-processing program (IMS Client V15Q4, Imagic, Switzerland). The empty shells were imaged using a camera (Canon EOS 50D) in a frontal, lateral, apical, and ventral position. The shell height and shell diameter were measured with perpendicular shell axis with the calliper.
Abbreviations used in the anatomical descriptions and figures:
At atrium
AG albumin gland
AS atrial stimulator
BC bursa copulatrix
BCD diverticulum of the bursa copulatrix
D shell diameter
DS dart sac
Ep epiphallus
Fl flagellum
FO free oviduct
H shell height
HD hermaphroditic duct
MG mucus glands
MRP musculus retractor penis
PA penial appendix
Pe penis
PF penial flap
PP1 proximal penial papilla
PP2 distal penial papilla
PS penis sheath
Va vagina
VD vas deferens
ResultsPhylogenetic results
The RAxML analysis of the concatenated data set (Fig. 2) recovered the genus Alabastrina as sister genus to Siretia and Otala. This node is supported with a ML support value of 90. The species A.tistutensis Galindo, 2018 clusters within the five specimens of A.alabastrites (Michaud, 1833). The monophyly of S.pallaryi (Kobelt, 1909) and Otala (and thus the separation of S.pallaryi and Alabastrina) is highly supported (bootstrap value of 99). The monophyly of Otala is not statistically supported (bootstrap value of 61). Within Otala we recovered three major clades, i.e., O.punctata (OF Müller, 1774), O.lactea (OF Müller, 1774), and O.xanthodon (Anton, 1838). The specimen of “Tingitanaminetteidecussata” (nomen nudum) clusters within the O.xanthodon clade. The monophyly of O.lactea is not statistically supported (bootstrap value of 65). Within O.xanthodon there are some nodes with very low support, especially the node which includes “Tingitanaminetteidecussata” (NMBE 549840). Otalal.murcica (Rossmässler, 1854) (NMBE-554175 and NMBE-554176 in Figs 2, 3) nests within the O.lactea clade. Both, the separate mitochondrial and nuclear tree show the same topology as the concatenated tree. They can be found in the supplementary material (Suppl. materials 1, 2).
Maximum Likelihood (RAxML) tree based on concatenated data set of COI, 16S, H3, and 5.8 S rRNA+ITS2. Numbers represent bootstrap support values from the ML analysis.
Bayesian Inference tree based on concatenated data set of COI, 16S, H3, and 5.8 S rRNA+ITS2. Numbers represent Bayesian posterior probabilities.
https://binary.pensoft.net/fig/299957
The Bayesian Inference analysis of the concatenated data set (Fig. 3) recovered the monophyly of Alabastrina. This node is statistically supported (posterior probability of 1). The monophyly of S.pallaryi and Otala and thus the separation of S.pallaryi and Alabastrina is fully supported. There is no difference in both types of analyses in the O.lactea and the O.xanthodon clade. The separate mitochondrial and nuclear trees can be found in the supplementary material (Suppl. materials 3, 4).
Taxonomic accounts
The nomenclature of the parts of the genital organs follows Neubert and Bank (2006) and Neubert (2014). In Table 3, the traits of the genital organs are summarised.
Traits of genital organs.
A.alabastrites
A.tistutensis
S.pallaryi
O.lactea
O.punctata
O.xanthodon
relative size of the AS
medium
medium
no data
large
large
large
penial flap
yes
yes
no data
no
no
no
relative size of the Fl
short
short
no data
long
medium
long
relationship BC:BCD
1:1
no data
no data
1.5:2
1:1
1.5:2
no. of penial papillae
1
1
no data
2
1
2
penial appendix
yes
yes
no data
no
no
no
Alabastrina Kobelt, 1904
1904 Alabastrina Kobelt, in Rossmässler: Iconographie der Land- & Süsswasser-Mollusken, (2) 11: 33, 132, 194 [type species Helixalabastrites Michaud, 1833 by OD].
1904 Alabastra Kobelt, in Rossmässler: Iconographie der Land- & Süsswasser-Mollusken, (2) 11: 100.
Currently, this genus is subdivided in six subgenera (Schileyko 2006). This system is more or less completely based on shell characters and only for a few specimens the morphology of the genital organs has been investigated and published. Schileyko (2006: 1794, fig. 2297B, C) shows the genital organs of Helixhieroglyphicula Michaud, 1833, which is the type species of Michaudia Pallary, 1926 [by original designation]. In his definition of Alabastrina sensu lato, he uses the character state “branches of mucus glands before entering common duct form distinct swellings” (Schileyko 2006: 1792). This interesting trait is not seen in any of the Alabastrina species investigated by us. Holyoak and Holyoak (2017: 426, Table 1) relegate Michaudia into the synonymy of Otala, also based on Schileyko’s figure arguing with the conformity in the structure of the interior of the proximal penis. The assumption by Schileyko (2006) that Alabastrina agrees with Otala on the presence of two penial papillae is wrong.
Without further comment, Holyoak and Holyoak (2017) consider Loxana Pallary, 1899 a separate genus, follow Razkin et al. (2015) in leaving Atlasica Pallary, 1917 as a subgenus of Alabastrina, and omit Lechatelieria Pallary, 1926. Taxon sampling in Razkin et al. (2015) is not sufficient enough to clearly reveal the subgeneric position of Atlasica. Based on our anatomical investigation, the genus Alabastrina can now be newly characterised using the following traits of the genital organs: Penis with a single penial papilla (PP) with a central pore, distal penis with penial flap (PF), proximal penis with a small penial appendix (PA); epiphallus and flagellum of similar length; mucus glands (MG) multifid, branches very long and slender.
Nomenclatural remark: Kobelt established the names Alabastra and Alabastrina simultaneously in the register volume of the “Iconographie”. In this work, he presented a register on the “System der palaearktischen Binnenconchylien”, listing a genus group name together with a single species group name (129 ff.). In the second register (171 ff.), he provided a systematically ordered list with information on all taxa ever published in the “Iconographie”, and affiliated these taxa into the new system as outlined before in register 1. Both registers are accompanied by text dealing with zoogeographic considerations and taxonomic remarks.
The name Alabastra was used three times exclusively on page 100 (in combination with a species list). The name Alabastrina was used on page 33 (zoogeographic context), page 132 (systematic register combined with the species group name alabastrites), page 158 (a list of potential members of Alabastrina including alabastrites), and finally page 194 (amended list of illustrated taxa of Alabastrina sensu Kobelt). According to ICZN 24.2.4 we deem Kobelt to act here as First Reviser, because he consequently used the name Alabastrina in his registers. We interpret the name Alabastra to constitute an erroneous misspelling.
Both genus group names included species lists of differing composition, the name alabastrites was always included (loc. cit.). In the first register, the name Alabastrina was combined with a single species (p. 132). We consider this act a designation of the type species by the original author (OD); Schileyko’s note on the type species selection (2006: 1792) as “monotypy” is erroneous.
Alabastrinaalabastrites (Michaud, 1833)
Figs 4–8
1833 Helixalabastrites Michaud, Catalogue des testacés vivans envoyés d’Alger par M. Rozet, capitaine au corps royal d’État-Major, au cabinet d’Histoire Naturelle de Strasbourg: 4, figs 6–8 [Oran].
1833 Helixsoluta Michaud, Catalogue des testacés vivans envoyés d’Alger par M. Rozet, capitaine au corps royal d’État-Major, au cabinet d’Histoire Naturelle de Strasbourg: 3, figs 9, 10 [Oran].
Type specimens: Helixalabastrites: syntype MHNL 45000690; Helixsoluta: syntype MHNL 45000679.
Specimens examined: for sequenced specimens, see Table 1.
Description. The range of the shell diameter of the investigated specimens is between 14.93–22.77 mm and shell height is between 10.85–13.45 mm. The shell of this species is pale and often with dark brown stripes. Some individuals do not show any stripes at all (Figs 4B, 6A). There is none to one tooth found in the aperture.
Alabastrina type specimens. AHelixsoluta, syntype MHNL 45000679, Oran, Algeria, coll. Michaud, D = 24.15 mm BHelixalabastrites, syntype MHNL 45000690, Oran, Algeria, coll. Michaud, D = 22.48 mm. All photographs by Kneubühler & Neubert, × 1.5.
https://binary.pensoft.net/fig/299958
This species has a rather short flagellum which is a bit shorter than the penis. MG are thin and fragile. The epiphallus goes over into the penial lumen without any penial papilla. Parallel but outside of the epiphallus is a penial appendix found. This penial appendix lies next to the epiphallus and is also covered by the penial sheath. It is blind on one side and opens into the penial lumen on the other side (PA in Fig. 5C, D). From there a huge penial papilla (PP) points towards the atrium. The PP is surrounded by massive muscles. In the atrium is a large atrial stimulator found and a smaller is located at the exit of the penis (PF).
Alabastrinaalabastrites (NMBE 549817), Kebdana Mountain, Morocco; A shell B situs C penis D penial lumen; D = 21.91 mm, H = 13.36 mm, situs length 27.57 mm (atrium-flagellum). All photographs by Kneubühler, shell × 1.5.
Alabastrinaalabastrites (NMBE 549812), cave Ifri n’Ammar, Morocco; A shell B situs C penis D penial lumen; D = 19.72 mm, H = 13.00 mm, situs length 26.27 mm (atrium-BCD). BC lost during dissection. All photographs by Kneubühler, shell × 1.5.
Alabastrinaalabastrites (NMBE 549813), hills El Batel, Morocco; A shell B situs C penis; D = 17.25 mm, H = 10.85 mm, situs length 13.46 mm (atrium-flagellum). Situs is not complete. All photographs by Kneubühler, shell × 1.5.
Alabastrinaalabastrites (NMBE 549816), Etsedda/ Kebdana, Morocco; A shell B situs C penis; D = 22.77 mm, H = 13.45 mm, situs length 36.84 mm (atrium-BCD). BC destroyed. All photographs by Kneubühler, shell × 1.5.
2018 Alabastrinatistutensis Galindo, Mostra mondiale, Cupra Marittima (2): 22–26.
Type specimen: Alabastrinatistutensis: holotype MMM Cupra Marittima (2): 23.
Specimens examined: for sequenced specimen, see Table 1.
Description. The shell is pale and characterised by a sharp keel. The aperture is white with a white lip. The mucus glands (MG) are fragile and slender. The flagellum is slightly shorter than the penis. The epiphallus is characterised by longitudinal tissue ridges and goes over into the penial lumen without any penial papilla. Parallel but outside of the epiphallus is a penial appendix found (PA in Fig. 9C). It is together with the epiphallus covered by the penial sheath. The PA is blind on one side and the other side opens into the penial lumen. This species possesses one penial papilla (PP in Fig. 9C) which is slightly smaller than in A.alabastrites but it is clearly visible. A large atrial stimulator is found in the atrium and a smaller stimulator is situated in front of the exit of the penis.
Alabastrinatistutensis (NMBE 555174), Tiztoutine, Morocco; A shell B situs C penis; D = 19.59 mm, H = 8.74 mm, situs length 13.51 mm (atrium-flagellum). Situs not complete. All photographs by Kneubühler, shell × 1.5.
1926 Siretia Pallary, Journal de Conchyliologie, 70: 19.
This genus is characterised by a triangular, toothless aperture, the short upper edge of the shell, its flat form, and by having four dark bands (Pallary 1926). Although Siretia has a peculiar shell morphology, Schileyko (2006) considers it as a subgenus of Alabastrina. Our phylogenetic analyses reveal it as a separate genus.
Siretiapallaryi (Kobelt, 1909)
Figure 10
1909 Archelixpallaryi Kobelt, Nachrichtsblatt der Deutschen Malakozoologischen Gesellschaft, 41 (3): 134 [Taforalt im Gebiet der Beni Snassen].
1914 Archelixpallaryi, – Kobelt: in Rossmässler: Iconographie der Europäischen Land- & Süsswasser-Mollusken (2) 20: 21, fig. 2790.
Type specimen: Siretiapallaryi: syntype SMF 75926.
Specimens examined: for sequenced specimen, see Table 1.
Description. In Figure 10B, a syntype of S.pallaryi from Teforalt (= Taforalt), Morocco (coll. CR Boettger ex Kobelt) is shown. The type specimen is slightly larger than our investigated specimen (Fig. 10A). Both show similar shell morphology and stripe pattern. Unfortunately, our specimen was badly preserved and a juvenile, therefore no proper investigation of the genital organs could be made.
ASiretiapallaryi (NMBE 549815), Kebdana Mountain, Morocco, D = 16.82 mm, H = 8.81 mm; BS.pallaryi (SMF 75926), Teforalt (= Taforalt), Morocco, coll. CR Boettger, D = 19.38 mm. All photographs by Kneubühler & Neubert, × 1.5.
https://binary.pensoft.net/fig/299964
Remarks. Holyoak and Holyoak (2017: 446) attribute this species to A Koch. However, in the description Kobelt explicitly mentions “Koch mss”. Therefore, Kobelt is considered the nomenclatural author of this taxon.
Otala Schumacher, 1817
1817 Otala Schumacher, Essai d’un nouveau système des habitations des vers testacés: 58, 191 [type species Helixlactea OF Müller, 1774, by subsequent designation Pilsbry, 1895: 323].
1904 Otala (Dupotetia) Kobelt: in Rossmässler: Iconographie der Europäischen Land- & Süsswasser-Mollusken (2) 11: 158 [type species Helixdupotetiana Terver, 1839 by original designation].
1918 Alabastrina (Tingitana) Pallary, Bulletin de la Société d’ Histoire naturelle de l’Afrique du Nord, 9 (7): 145 [type species Archelixminettei Pallary, 1917 by monotypy].
This genus was recently revised by Holyoak and Holyoak (2017). After examining several hundreds of specimens from Morocco and Algeria, they distinguish five species within the genus Otala, i.e., O.punctata, O.lactea, O.xanthodon, O.tingitana (Paladilhe, 1875), and O.hieroglyphicula (Michaud, 1833). The species formerly attributed to Tingitana Pallary, 1918, and Dupotetia Kobelt, 1904 (genera which appeared to have species in the area of the Kebdana) are now lumped under Otalatingitana. This lumping approach is supported by the molecular study of Helicoidea by Razkin et al. (2015), who revealed that the genus Tingitana is nested within Otala. In our phylogenetic analysis we included a specimen of the well-known shell form “Tingitanaminetteidecussata”, which clustered within the O.xanthodon clade thus supporting the results of Razkin et al. (2015) and Holyoak and Holyoak (2017). More taxon sampling is needed to reveal the phylogenetic relationships within Otala.
Otalalactea (OF Müller, 1774)
Figs 11–16
Type specimens: Helixlucasii: MNHN IM-2000-31721.
Specimens examined: for sequenced specimens, see Table 1.
Description. The shell of O.lactea is characterized by a dark aperture. The shell diameter of the investigated specimens ranges between 27.01–40.81 mm and shell height between 15.77–21.75 mm. This species has a large and thick penial tube. It has two distinct penial papillae with each a large central pore. The distal penial lumen between the large tongue-shaped atrial stimulator and the distal penial papilla (PP2) exhibits longitudinal ridges. The distal penial papilla is located ca. 2 mm distally to the atrium. The penial chamber which is bordered by the two penial papillae ranges between 2–4 mm and is characterised by strong annular tissue folds. There is a short transformation zone between the proximal penial papilla (PP1) and the epiphallus. The epiphallus is characterised by longitudinal tissue ridges. The flagellum is ca. 1.5× the length of the penis. The BCD is ca. double in length as the BC, except for the specimen in Figure 13, where they are approximately the same length. The vagina is stout and short. The MG consist of two massive stems which subdivide into ten smaller branches.
Otalalactea (NMBE 553246), W Almocageme, Portugal; A shell B situs C penis and atrium; D = 29.82 mm, H = 18.71 mm, situs length 41.34 mm (atrium-BCD). All photographs by Kneubühler, shell × 1.5.
Otalalactea (NMBE 554174), N Málaga, Spain; A shell B situs; D = 27.25 mm, H = 18.60 mm, situs length 22.90 mm (atrium-albumin gland); juvenile, BC destroyed. All photographs by Kneubühler, shell × 1.5.
Otalalactea (NMBE 555171), Hassi Ouenzga/ Oriental, Morocco; A shell B situs C penis and atrium; D = 22.63 mm, H = 14.85 mm, situs length 34.60 mm (atrium-flagellum). All photographs by Kneubühler, shell × 1.5.
Otalalactea (NMBE 549951), W Aoulouz, Morocco; A shell B situs; D = 27.01 mm, H = 15.77 mm, situs length 30.77 mm (atrium-flagellum); juvenile; situs not complete; BC destroyed. All photographs by Kneubühler, shell × 1.5.
https://binary.pensoft.net/fig/299968
Remarks. The analysis includes also specimens of O.l.murcica (Fig. 15) from Almería, Spain, which is the type locality. This taxon is characterised by a larger shell and an aperture, which is enlarged and more reflected (Cadevall and Orozco 2016). The morphology of the genital organs shows no difference to the specimens of O.lactea investigated from Portugal or Morocco.
Otalalactea (NMBE 554175), W Almería, Spain; A shell B situs C penis and atrium; D = 31.89 mm, H = 18.23 mm, situs length 57.86 mm (atrium-BCD). All photographs by Kneubühler, shell × 1.5.
https://binary.pensoft.net/fig/299969
In a small area in north-eastern Morocco, another form of O.lactea occurs, namely Helixlucasii (Fig. 16D). Our investigation of a specimen from this population revealed some differences in the anatomy of the genital organs (Fig. 16C). The penial chamber is much longer than in the other specimens of O.lactea. The length of the penial chamber (PP1-PP2) is 4 mm and the length of the distal penial lumen (PP2-AS) is 1.8 mm. The internal structures differ substantially. Here, the inner walls of this tube are filled by numerous fine transverse ridges arranged in a very dense annular pattern. All other specimens seen so far displayed an irregular network of tissue folds in this section of the penis. Additionally the shell is quite large and flat with a comparatively strong basal tooth or strengthened lip.
Otalalactea (NMBE 545594); Etsedda/Kebdana, Morocco; A shell B situs C penis; D = 40.81 mm, H = 21.75 mm, situs length 61.47 mm (atrium-BCD), BC destroyed; DH.lucasii (syntype MNHN IM-2000-31721), Oran, Algeria, D = 35.4 mm. All photographs by Kneubühler & Neubert, shell original size.
Specimens examined: for sequenced specimens, see Table 1.
Description. The shell is characterized by a white lip and a basal tooth. This species is characterized by a long and thick penial tube. It has a large penial papilla (PP), which is located ca. 2 mm distally to the atrium (Figs 17, 18) with a large central pore. The second proximal penial papilla is reduced and inconspicuous. The distal penial lumen between the atrial stimulator and the penial papilla exhibits a few low longitudinal ridges intersected by many small annular folds. The proximal lumen between penial papilla and epiphallus is filled by a network of irregularly shaped folds and small and large ridges. The epiphallus is characterised by longitudinal tissue ridges with a small transformation zone at the proximal end of the penial lumen. The flagellum has approximately the same length as the penis. The vagina is short and stout. The mucus glands (MG) consist of two massive stems which subdivide into 10–12 smaller subsequent branches. The BCD has approximately the same length as the BC. They are ca. 3× the length of the flagellum and the penis. The dominant structure in the atrium is a large, folded stimulator, which was also mentioned by De Mattia and Mascia (2011).
Otalapunctata (NMBE 534228); Makouda, Algeria; A shell B situs C penis; D = 36.02 mm, H = 22.37 mm, situs length 59.77 mm (atrium-BCD). All photographs by Kneubühler, shell original size
Otalapunctata (NMBE 554171); W Málaga, Spain; A shell B situs C penis; D = 30.30 mm, H = 18.05 mm, situs length 70.10 mm (atrium-BC). All photographs by Kneubühler, shell × 1.5.
Specimens examined: for sequenced specimens, see Table 1.
Description. The shell is characterized by a dark aperture with a white and strongly reverted lip. This species possesses one basal tooth. A palatal tooth is found in some specimens. The shell diameters of the investigated specimens range between 21.47–27.77 mm and shell height between 13.37–16.04 mm. Otalaxanthodon has two distinct penial papillae with each a large central pore. The distal penial lumen between the atrial stimulator and the distal penial papilla (PP2) exhibits smooth longitudinal tissue ridges. The penial chamber which is bordered by the two penial papillae is filled by a network of irregularly shaped tissue folds and is ca. 3 mm long. There is a short transformation zone between the proximal penial papilla (PP1) and the epiphallus. The epiphallus contains few smooth longitudinal ridges. This species has a large flagellum which is ca. double the length of the penis. The BC is a thin tube and ca. half the length of the BCD. It has two massive mucus glands (MG) which subdivide in four thinner branches of which each again subdivides in two thin branches. The dominant structure in the atrium is a large tongue-shaped stimulator.
Otalaxanthodon (NMBE 549825), Moulouya, Morocco; A shell B situs C atrium and penis; D = 23.13 mm, H = 14.49 mm, situs length 32.68 mm (atrium-BCD). BC destroyed. All photographs by Kneubühler, shell × 1.5.
Otalaxanthodon (NMBE 549826), Moulouya, Morocco; A shell B situs C atrium and PP2 D penial chamber; D = 21.47 mm, H = 14.22 mm, situs length 37.23 mm (atrium-BCD). All photographs by Kneubühler, shell × 1.5.
Otalaxanthodon (NMBE 549841), Kebdana Mountain, Morocco; A shell B situs; D = 26.85 mm, H = 15.74 mm, situs length 33.58 mm (atrium-flagellum). Situs not complete. All photographs by Kneubühler, shell × 1.5.
Otalaxanthodon, Kebdana, Moulouya valley, Morocco; A shell from O.xanthodon (NMBE 555169), D = 22.33 mm, H = 13.61 mm; B shell from O.xanthodon (NMBE 555170), D = 23.10 mm, H = 13.37 mm. Kebdana, Djebel Sebaa Reyal/ Rif C shell from O.xanthodon (NMBE 549843), D = 27.77 mm, H = 16.04 mm. All photographs by Kneubühler, shell × 1.5.
Otalaxanthodon (NMBE 549820), Guercif, Morocco; A shell B situs C atrium and penis; D = 26.74 mm, H = 15.96 mm, situs length 42.81 mm (atrium-BCD), BC destroyed; DHelixzaffarina Terver, 1839 (syntype MHNL 45001034), Oran, Algeria, coll. Michaud, D = 29.54 mm. All photographs by Kneubühler & Neubert, shell × 1.5.
Specimens examined: Otalatingitana (NMBE 510549); for the sequenced specimen of “Tingitanaminetteidecussata” NMBE 549840, see Table 1.
Nomenclatorial note: The name “decussata Pallary” is a nomen nudum as already stated by Holyoak and Holyoak (2017: 463). Pallary never made the name available, nor did Llabador (1952). For the latter publication, the provisions of Article 13 ICZN (names published after 1930) rule that every new name must “be accompanied by a description or definition that states in words characters that are purported to differentiate the taxon” or Article 13.1.2. “be accompanied by a bibliographic reference to such a published statement”. No such statements are provided by Llabador. This taxon is well known and often treated as a subspecies of Tingitanaminettei (Pallary, 1917) (see for example Cossignani 2014). The genus Tingitana Pallary, 1918 is synonymised with Otala by Holyoak and Holyoak (2017).
Description. The shells of “decussata” are flat and have a sharply keeled last whorl. The aperture is oval and dark brown inside with a white lip and a strong basal tooth. “Tingitanaminetteidecussata” has a network-like sculpture on its surface (Fig. 25). This is in contrast to Otalatingitana with a rather smooth surface and a few weakly developed radial ribs. In this species, the interior of the aperture is brighter and the basal tooth conspicuously smaller. Typically, O.xanthodon has a smooth shell with evenly rounded whorls and up to three apertural denticles.
Otalatingitana (NMBE 510549), Tarzout de Guigou, Morocco, D = 27.42 mm, H = 14.38 mm (specimens from the type locality of Archelixminettei Pallary, 1917). All photographs by Kneubühler, shell × 1.5.
“Tingitanaminetteidecussata” (NMBE 549840), Kebdana, Morocco; A shell B situs C atrium and penis; D = 32.45 mm, H = 16.12 mm, situs length 28.84 mm (atrium-flagellum). Situs not complete. All photographs by Kneubühler, shell × 1.5.
https://binary.pensoft.net/fig/299975
The genital organs of “decussata” are characterised by two distinct penial papillae, each with a central pore. The distal penial lumen between the atrial stimulator and the distal penial papilla (PP2) is characterised by a network of irregularly shaped folds with large and small ridges. The penial chamber exhibits many annular tissue folds and is ca. 3 mm long. Between the proximal penial papilla (PP1) and the epiphallus is a short transformation zone. The epiphallus is characterised by two strong and several smooth longitudinal ridges. The mucus glands consist of two massive stems which subdivide into several thinner branches which again become thinner in the second half. The dominant structure in the atrium is a large tongue-shaped stimulator. There are almost no differences in the inner and outer morphology of the genital organs of “decussata” and O.xanthodon specimens.
Remarks. According to field observations by R Hutterer, this particular taxon does only occur on top of one mountain in the Kebdana range; comparison with similar specimens illustrated by Cossignani (2014: 109) from Ras el Ma and Tazouta is pending. The distribution area of O.tingitana/minettei is far and separated by lowlands, so a position of this taxon as a species in its own right is highly probable. However, as long as topotypic specimens of O.tingitana are missing in the genetic analysis, the exact taxonomic position of “decussata Pallary” remains unclear. Our results signal a position within or close to O.xanthodon rather than to O.tingitana.
Discussion
The results of our study strongly support the monophyly of the genera Alabastrina and Otala within the tribe Otalini. Alabastrinaalabastrites is morphologically as well as genetically clearly separated from the genera Siretia and Otala. All investigated specimens within Alabastrina show the unique trait of the presence of a blind penial appendix. This is an anatomical character, which has never been reported before within the Helicidae. The function of this penial appendix is not known. Schileyko’s system which was based on morphology only, is incorrect as we could demonstrate in our phylogeny that the species Archelixpallaryi Kobelt, 1909, which is the type species for the genus Siretia, clusters outside the Alabastrina clade. We consider this taxon as a separate genus. Anatomical and genetic data for Helixbailloni Kobelt, 1888, the type species of Guilia Pallary, 1926 also suggest a phylogenetically separate position of this genus (Kneubühler et al. in prep.). The position of A.tistutensis within the clade of A.alabastrites shows that this extreme local shell form should probably be considered a local subspecies rather than a species in its own rights. Further sampling is necessary to resolve the problem.
The phylogenetic results clearly show that Siretia is separated from Alabastrina. In the ML analyses Siretia forms a lineage separate from Otala (Fig. 2; Suppl. materials 1, 2). However, in the Bayesian Inference analyses, Siretia clusters within the Otala clade (Fig. 3; Suppl. materials 3, 4). Thus, the monophyly of Otala is not supported. It cannot be excluded that Siretia forms a subgenus or even a synonym of Otala. Unfortunately, we cannot present anatomical data for S.pallaryi because of the bad preservation of the only specimen we could analyse. More sequence data are necessary to corroborate the monophyly of Otala and to resolve the relationships within the Otala clade (including Siretia). For the time being, Siretia is considered here as a separate unit because of the differences in shell shape. Holyoak and Holyoak (2017: 423) regard Siretia as a distinct genus within the Otalini.
Otalalactea is characterized by a dark aperture, which clearly differentiates it from O.punctata with a white aperture. We investigated several populations of O.lactea from Morocco, Spain and Portugal and they all cluster together in the phylogenetic analysis. Hesse’s (1911) investigations of the outer morphology of the genital organs of Archelixpunctata, A.lactea, and A.lucasi showed no difference to our results. In contrast to Holyoak and Holyoak (2017), we could distinguish the species O.lactea and O.punctata without any doubt by their genital anatomy. Otalapunctata has one strongly developed penial papilla and a second which is nearly completely reduced, whereas O.lactea has two massive and distinct penial papillae. Unfortunately, Holyoak and Holyoak (2017: 425, Table 1) do not describe the form of the proximal verge (PP1 herein) for each species nor do they provide a drawing. This hampers the interpretation of the data known so far and we agree that more detailed study may be necessary for a reliable comparison of species.
We also investigated specimens of O.l.murcica from Almería, Spain; from a genetic point of view there is no difference to the remaining specimens of O.l.lactea. The two specimens of O.l.murcica included in the analyses from the same population (NMBE-554175 and NMBE-554176 in Figs 2, 3) cluster together with the Portuguese specimen of O.lactea, which originates close to the type locality of the neotype of O.lactea designated by Holyoak and Holyoak (2017: 446). For this reason we conclude that this subspecies has to be considered a synonym of O.lactea.
The specimen from Etsedda, Morocco (NMBE-545594 in Figs 2, 3) clusters as the sister lineage of all investigated O.lactea specimens. It shows a slightly different shell morphology and genital anatomy (Fig. 16A, B, C). The shells of this population strongly resemble Helixlucasii (syntype shown under Fig. 16D). However, the bootstrap support value for this clade (65) is too low to currently allow the separation as a distinct species or whether it falls within the range of variability of O.lactea. More specimens are needed here to corroborate the differences in the anatomical details of the genital organs as well as the separate position on the phylogeny.
“Tingitanaminetteidecussata” clusters within the specimens of O.xanthodon but with a low support (Figs 2, 3). The genital organs show strong similarities to other O.xanthodon specimens as exemplified by the system of two penial papillae, the short penial chamber, the massive mucus glands, and the large atrial stimulator. However, the shell morphology of this form is clearly different. This could be due to a local adaptation to a rocky habitat since the gastropod shell form is strongly influenced by the substrate the species live on (Goodfriend 1986); specimens with a flat shell can hide more easily in crevices, particularly in limestone. This conflicts with the definition of Tingitana by Pallary, who erected this genus for species with a keeled shell. Next to the observation cited above that keeled shells are probably an adaptation to a rocky environment with crevices, juvenile shells of large helicid species often show this phenomenon of a keeled shell (see for example species of Levantina Kobelt, 1871, Codringtonia Kobelt, 1898, Isaurica Kobelt, 1901, etc. (Holyoak and Holyoak 2017)). Consequently, this trait is unsuitable for generic definition; its use even for species delimitation is disputable.
Holyoak and Holyoak (2017) synonymised H.zaffarina (a species usually under Dupotetia) with O.xanthodon. Therefore, we included a specimen that usually would have been identified as D.zaffarina in our study (Fig. 23A), and compared the shell with that of the syntype (Fig. 23D). We agree here with the synonymisation of H.zaffarina with O.xanthodon, because our genetic analyses revealed that this specimen clusters within the specimens of O.xanthodon.
Acknowledgements
We are grateful to Estée Bochud (NMBE Switzerland) for support during the anatomical research and technical advice, José Ahuir Galindo (Málaga, Spain) and Adrienne Jochum (NMBE Switzerland) for providing specimens used in this study. RH is grateful to the support of Deutsches Archäologisches Institut (Bonn) and the Institut National des Sciences de l’Archéologie et du Patrimoine (Rabat). We acknowledge the valuable comments of the reviewers B Hausdorf and MT Neiber (both Hamburg) on an earlier draft of this paper.
ReferencesAltekarGDwarkadasSHuelsenbeckJPRonquistF (2004) Parallel Metropolis-coupled Markov chain Monte Carlo for Bayesian phylogenetic inference.20: 407–415. https://doi.org/10.1093/bioinformatics/btg427AntonHE (1838) Eduard Anton, Halle, 110 pp. https://doi.org/10.5962/bhl.title.11509Bouaziz-YahiateneHPfarrerBMedjdoub-BensaadFNeubertE (2017) Revision of Massylaea Möllendorff, 1898 (Stylommatophora, Helicidae).694: 109–133. https://doi.org/10.3897/zookeys.694.15001CadahiaLHarlJDudaMSattmannHKruckenhauserLFeherZZoppLHaringE (2014) New data on the phylogeny of Ariantinae (Pulmonata, Helicidae) and the systematic position of Cylindrusobtusus based on nuclear and mitochondrial DNA marker sequences.52(2): 163–169. https://doi.org/10.1111/jzs.12044CadevallJOrozcoA (2016) Caracoles y babosas de la península Ibérica y Baleares. Nueva guías de campo. Omega, Barcelona.ColganDMcLauchlanAWilsonGDFLivingstonSPEdgecombeGD (1998) Histone H3 and U2 snRNA DNA sequences and arthropod molecular evolution.46: 419–437. https://doi.org/10.1071/ZO98048CossignaniT (2014) African Landshells. L’informatore Piceno, Ancona.De MattiaWMasciaF (2011) Otalapunctata (O.F. Müller, 1774) (Stylommatophora: Helicidae) in Italy.29(1): 39–46.Férussac D’AudebardAÉJPDeshayesGP (1819–1851) J-B Baillière, Paris, 184 pp. [T. 1], 402 pp [T. 2(1)], 1–260, 1–22 [2(2)]. [Atlas 1: 70 pls; Atlas 2: 166 + 5 pls]FolmerOBlackMHoeWLutzRVrijenhoekR (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates.3(5): 294–299.GalindoJA (2018) Three new terrestrial gastropod species and two new subspecies from Morocco.2: 22–26.GenBank (2018) National Center for Biotechnology Information, U.S. National Library of Medicine, Rockville Pike. https://www.ncbi.nlm.nih.gov/genbank/ [last accessed 17.12.2018]GoodfriendGA (1986) Variation in land-snail shell form and size and its causes: a review.35(2): 204–223. https://doi.org/10.2307/2413431HesseP (1907–1920) Subfamilia Helicinae. In: Iconographie der Land- & Süsswasser-Mollusken mit vorzüglicher Berücksichtigung der europäischen noch nicht abgebildeten Arten (2) 14 (1/2): 1–64, pl. 361–370 (13 February 1907); (2) 14 (3/4): 65–128, pl. 371–380 (2 September 1907); (2) 14 (5/6): 129–172 + VII pp., pl. 381–390 (12 September 1908); (2) 16 (1/2): 1–42, pl. 421–430 (23 June 1909); (2) 16 (3/4): 43–66, pl. 431–440 (6 June 1910); (2) 16 (5/6): 67–119 + 5 pp., pl. 441–450 (3 August 1911); (2) 23 (1/2): 1–72, pl. 631–640 (October 1915); (2) 23 (3/4): 73–152, pl. 641–650 (11 November 1919); (2) 23 (5/6): 153–262 + 5 pp., pl. 651–660 (24 November 1920). CW Kreidel, Wiesbaden. https://doi.org/10.1016/S0033-3506(09)80188-1HolyoakDTHolyoakGA (2017) A revision of the land-snail genera Otala and Eobania (Gastropoda, Helicidae) in Morocco and Algeria.42(6): 419–490.HuelsenbeckJPRonquistF (2001) MRBAYES: Bayesian inference of phylogeny.17: 754–755. https://doi.org/10.1093/bioinformatics/17.8.754HuttererRMikdadARipkenTEJ (2011a) Species composition and human exploitation of terrestrial gastropods from Taghit Haddouch, an Early Holocene archaeological site in NE Morocco.140: 57–78. https://doi.org/10.1127/arch.moll/1869-0963/140/057-075HuttererREiwangerJLinstädterJMikdadA (2011b) Konsum von Landschnecken im Neolithikum: Neue Daten aus dem östlichen Rif (Marokko).8: 29–34.HuttererRLinstädterJEiwangerJMikdadA (2014) Human manipulation of terrestrial gastropods in Neolithic culture groups of NE Morocco.320: 83–91. https://doi.org/10.1016/j.quaint.2013.12.006KobeltW (1887–1888) . In: Rossmässler EA (Ed.) Iconographie der Land- & Süßwasser-Mollusken mit vorzüglicher Berücksichtigung der europäischen noch nicht abgebildeten Arten (2) 3 (1/2): 1–12, pl. 61–70 [15. Apr. 1887]; (2) 3 (3/4): 13–36, pl. 71–80 [6. Oct. 1887]; (2) 3 (5/6): 37–60, pl. 81–90 [10. March 1888]. Kreidel, Wiesbaden.KobeltW (1901–1904) Iconographie der Land- & Süsswasser-Mollusken mit vorzüglicher Berücksichtigung der europäischen noch nicht abgebildeten Arten von EA Rossmässler (2) 11: 80–264.KobeltW (1909) Diagnose einer neuen Archelix.41(3): 134–135.KobeltW (1914) Rossmässler’s Iconographie der Europäischen Land- & Süsswasser-Mollusken. (2) 20, 1–24. [pl. 541–552]LanfearRFrandsenPBWrightAMSenfeldTCalcottB (2016) PartitionFinder 2: new methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses. Molecular Biology and Evolution. https://doi.org/10.1093/molbev/msw260LlabadorF (1952) Contribution à l’étude de la faune malacologique terrestre et fluviatile du Rif oriental.92: 93–142. [pl 5]MichaudALG (1833) Catalogue des testacés vivans envoyés d’Alger, par M. Rozet (capitaine au corps royal d’état-major), au cabinet d’histoire naturelle des Strasbourg; notice présentée à la Société d’histoire naturelle de la même ville. Mémoires de la Société d’histoire naturelle de Strasbourg, 1–22.MikdadAEiwangerJAtkiHBen-NcerABokbotYHuttererRLindstädterJMoucineT (2000) Recherches préhistoriques et protohistoriques dans le Rif Oriental (Maroc).20: 109–167.MüllerOF (1774) Vermium terrestrium et fluviatilium, seu animalium infusoriorum, Helminthicorum et Testaceorum, non marinorum succincta historia. 2. Havinae & Lipsiae, I–XXXVI [= 1–36] + 1–214. https://doi.org/10.5962/bhl.title.46299NeiberMTHausdorfB (2015) Molecular phylogeny reveals the polyphyly of the snail genus Cepaea (Gastropoda: Helicidae).93: 143–149. https://doi.org/10.1016/j.ympev.2015.07.022NeubertE (2014) Revision of Helix (Linnaeus, 1758) in its eastern Mediterranean distribution area, and reassignment of Helixgodetiana (Kobelt, 1878) to Maltzanella (Hesse, 1917) (Gastropoda, Pulmonata, Helicidae).26: 1–200.NeubertEBankRA (2006) Notes on the species of Caucasotachea C Boettger, 1909 und Lindholmia P Hesse, 1919, with annotations to the Helicidae (Gastropoda: Stylommatophora: Helicidae).135(1): 101–132. https://doi.org/10.1127/arch.moll/0003-9284/135/101-132NeubertEKorábekO (2015) Comment on Psonis et al. (2015) Evaluation of the taxonomy of Helixcincta (Muller, 1774) and Helixnucula (Mousson, 1854); insights using mitochondrial DNA sequence data.49: 383–392. https://doi.org/10.1080/00222933.2015.1021874PaladilheA (1875) Étude sur les coquilles terrestres et fluviatiles rapportées du Maroc par le Dr. Bleicher. Revue et Magasin de Zoologie, pure et appliquée (3eme sér.3: 75–101. [pl 9]PallaryPM (1899) Deuxième contribution à l’étude de la faune malacologique du Nord-Ouest de l’Afrique. Supplement à “La faune malacologique du Maroc” de A Morelet. Journal de Conchyliologie 46(2) [1898]: 49–170. [pls 5–9; Paris] [9 February].PallaryPM (1917) Hélicidées nouvelle du Maroc.63(2): 126–141. [pl 5]PallaryPM (1918) Diagnoses d’une cinquantaine de mollusques terrestres nouveaux du Nord de l’Afrique.9(7): 137–152.PallaryPM (1926) Compléments à la faune malacologique de la Berbérie.70(1): 1–50. [pls 1–8]PallaryPM (1936) Deuxième complément a la faune malacologique de la Berbérie. Journal de Conchyliologie, 1er Trimestre, 32–39.PilsbryHA (1895) Manual of Conchology, structural and systematic, with illustrations of the species. By George W Tryon, Jr. Second series: Pulmonata, vol 9. (Helicidæ, vol. 7), part 36, frontispiece. Guide to the study of Helices. Conchological Section, Academy of Natural Sciences of Philadelphia, Philadelphia, 161–366. [pls 41–71]PfefferG (1930) Zur Kenntnis tertiärer Landschnecken. Geologische und Paläontologische Abhandlungen, Jena (NF) 17(3): 3–230 (153–380).PsonisNVardinoyannisKMylonasMPoulakakisN (2013) Evaluation of the taxonomy of Helixcincta (Muller, 1774) and Helixnucula (Mousson, 1854); insights using mitochondrial DNA sequence data.49: 383–392. https://doi.org/10.1080/00222933.2013.825023RazkinOGómez-MolinerBJPrietoCEMartínez-OrtíAArrébolaRMuñozBChuecaLJMadeiraMJ (2015) Molecular phylogeny of the western Palaearctic Helicoidea (Gastropoda, Stylommatophora).83: 99–117. https://doi.org/10.1016/j.ympev.2014.11.014RonquistFHuelsenbeckJP (2003) MRBAYES 3: Bayesian phylogenetic inference under mixed models.19: 1572–1574. https://doi.org/10.1093/bioinformatics/btg180RossmässlerEA (1854) Band 3, Heft 1–2, 139 pp. [pl 61–88]RourEChahlaouiAvan GoethemJL (2002) Etat actuel des connaissances de la malacofaune terrestre du Maroc.72: 189–199.SchileykoAA (2006) Treatise on recent terrestrial pulmonate molluscs. Part 13. Helicidae, Pleurodontidae, Polygyridae, Ammonitellidae, Oreohelicidae, Thysanophoridae. Ruthenica, Supplement 2, Moscow, 1765–1906.SchumacherCF (1817) Essai d’un nouveau système des habitations des vers testacés. Schultz, Copenhagen, [1–3], 1–287. [pls 1–22]SimonCFratiFBeckenbachACrespiBLiuHFlookP (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers.87(6): 651–701. https://doi.org/10.1093/aesa/87.6.651StamatakisA (2006) RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models.22(21): 2688–2690. https://doi.org/10.1093/bioinformatics/btl446TerverAP (1839) Catalogue des mollusques terrestres et fluviatiles, observés dans les possessions françaises au nord de l’Afrique, publié par M Terver. J-B Baillière, Paris & Savy, Lyon, 1–40. [pls 1–4]WaltherFNeiberMTHausdorfB (2016) Species complex or complex species? Integrative taxonomy of the land snail genus Rossmaessleria (Gastropoda, Helicidae) from Morocco and Gibraltar.14: 394–416. https://doi.org/10.1080/14772000.2016.1150905Supplementary materials10.3897/zookeys.843.32867.suppl13486692E83C81A8-D705-5934-AA0A-EAF9AD924F42
Maximum Likelihood (RAxML) tree based on mitochondrial data set of COI and 16S
PDF file
Numbers represent bootstrap support values from the ML analysis.
https://binary.pensoft.net/file/299976This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.Jeannette Kneubühler, Rainer Hutterer, Beat Pfarrer, Eike Neubert10.3897/zookeys.843.32867.suppl23486694AA306969-A1C6-5363-B048-7987376711BF
Maximum Likelihood (RAxML) tree based on nuclear data set of H3 and 5.8 S rRNA+ITS2
PDF file
Numbers represent bootstrap support values from the ML analysis.
https://binary.pensoft.net/file/299977This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.Jeannette Kneubühler, Rainer Hutterer, Beat Pfarrer, Eike Neubert10.3897/zookeys.843.32867.suppl334866963E75CB3E-D523-596A-B9F1-4E3D93B2F492
Bayesian Inference tree based on mitochondrial data set of COI and 16S
https://binary.pensoft.net/file/299978This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.Jeannette Kneubühler, Rainer Hutterer, Beat Pfarrer, Eike Neubert10.3897/zookeys.843.32867.suppl4348669811855DA8-AE8D-570D-A311-F1E768773E24
Bayesian Inference tree based on nuclear data set of H3 and 5.8 S rRNA+ITS2
https://binary.pensoft.net/file/299979This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.Jeannette Kneubühler, Rainer Hutterer, Beat Pfarrer, Eike Neubert