Next step in Monachacantiana (Montagu, 1803) phylogeography: northern French and Dutch populations (Eupulmonata, Stylommatophora, Hygromiidae).

Features of shell and genitalia as well as nucleotide sequences of selected mitochondrial and nuclear genes of specimens of Monachacantiana from ten northern French and two Dutch populations were compared with the same features of British and Italian populations. They were found to be very similar to populations previously identified as belonging to the CAN-1 lineage of M.cantiana. This confirms previous suggestions that M.cantiana was introduced to western Europe (England, France and the Netherlands) in historical times.

Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach.

Specimens obtained from ten populations of a Monacha species from the central Apennines were compared with six molecular lineages of Monacha cantiana s. l. (CAN-1, CAN-2, CAN-3, CAN-4, CAN-5, CAN-6) and two other Monacha species (M. cartusiana and M. parumcincta), treated as outgroup, by molecular (nucleotide sequences of two mitochondrial COI and 16S rDNA as well as two nuclear ITS2 and H3 gene fragments) and morphological (shell and genital anatomy) analysis. The results strongly suggest that these populations represent a separate species for which two names are available: the older Helix pantanellii De Stefani, 1879 and the junior M. ruffoi Giusti, 1973. The nucleotide sequences created well separated clades on each phylogenetic tree. Genital anatomy included several distinctive features concerning vaginal appendix, penis, penial papilla and flagellum; instead, shell characters only enabled them to be distinguished from M. cartusiana and M. parumcincta. Remarkably, populations of M. pantanellii show high morphological variability. Shell variability mainly concerns size, some populations having very small dimensions. Genital variability shows a more intricate pattern of all anatomical parts, being higher as regards the vagina and vaginal appendix. Despite this morphological variability, the K2P distance range of COI sequences between populations is narrow (0.2-4.5%), if we consider all but three of the 53 sequences obtained. This research confirmed that the species of Monacha and their molecularly distinguished lineages can only occasionally be recognised morphologically and that they have significant inter- and intra-population variability. The possibility of using an overall approach, including shell, genital and molecular evidence, was taken in order to establish a reliable taxonomic setting.

Exploration of phylogeography of Monachacantiana s.l. continues: the populations of the Apuan Alps (NW Tuscany, Italy) (Eupulmonata, Stylommatophora, Hygromiidae).

Two new lineages CAN-5 and CAN-6 were recognised in four populations of Monachacantiana (Montagu, 1803) s.l. from the Italian Apuan Alps by joint molecular and morphological analysis. They are different from other M.cantiana lineages known from English, Italian, Austrian and French populations, i.e. CAN-1, CAN-2, CAN-3 and CAN-4, as well as from the other Italian Monacha species used for comparisons (M.parumcincta and M.cartusiana). Although a definite taxonomic and nomenclatural setting seems to be premature, we suggest that the name or names for these new lineages as one or two species should be found among 19th century names (Helixsobara Mabille, 1881, H.ardesa Mabille, 1881, H.apuanica Mabille, 1881, H.carfaniensis De Stefani, 1883 and H.spallanzanii De Stefani, 1884).

Exploring Monacha cantiana (Montagu, 1803) phylogeography: cryptic lineages and new insights into the origin of the English populations (Eupulmonata, Stylommatophora, Hygromiidae).

Molecular analysis of nucleotide sequences of mitochondrial cytochrome oxidase subunit 1 (COI) and 16S ribosomal DNA (16SrDNA) as well as nuclear histone 3 (H3) and internal transcribed spacer 2 of rDNA (ITS2) gene fragments together with morphological analysis of shell and genitalia features showed that English, French and Italian populations usually assigned to Monacha cantiana consist of four distinct lineages (CAN-1, CAN-2, CAN-3, CAN-4). One of these lineages (CAN-1) included most of the UK (five sites) and Italian (five sites) populations examined. Three other lineages represented populations from two sites in northern Italy (CAN-2), three sites in northern Italy and Austria (CAN-3), and two sites in south-eastern France (CAN-4). The taxonomic and nomenclatural setting is only currently available for lineages CAN-1 and CAN-4; a definitive frame for the other two requires much more research. The lineage CAN-1 corresponds to the true M. cantiana (Montagu, 1803) because it is the only one that includes topotypical English populations. The relationships and genetic distances support the hypothesis of the Italian origin of this lineage which was probably introduced to England by the Romans. The lineage CAN-4 is attributed to M. cemenelea (Risso, 1826), for which a neotype has been designated and deposited. Its diagnostic sequences of COI, 16SrDNA, H3 and ITS2 genes have also been deposited in GenBank. Molecular and morphological (shell and genitalia) features showed that M. parumcincta (Rossmässler, 1834) is a distinct taxon from the M. cantiana lineages.

Historical biogeography of Tyrrhenian land snails: the Marmorana-Tyrrheniberus radiation (Pulmonata, Helicidae).

The few studies available on Tyrrhenian land snails support high diversification in the Italian Peninsula and groups structured mainly by vicariant events. Here we investigated the phylogeny of a conchologically diversified group of Tyrrhenian land snails assigned to the genera Marmorana and Tyrrheniberus. We constructed a molecular phylogeny by sequencing two commonly used mtDNA genes (cytochrome oxidase I and the large ribosomal subunit). We also carried out conchological and anatomical analysis. Morphological (shell and genitalia) and genetic data (mitochondrial genes) showed paraphyly of Marmorana. Plio-Pleistocene events in the Tyrrhenian area may have structured relationships between species at regional scale while isolation by distance probably played a role in diversification between populations on a local scale. Continental populations experienced dispersal during interglacial periods and fragmentation and reduction during the dry cold climatic phases. Areas inhabited by Apennine Marmorana could represent relict and/or glacial refugia, with extinction in some areas along the Apennines and survival in the south. The results support a reassessment of taxonomy both at genus and species level and call for further analysis.

Molecular phylogeny and historical biogeography of the land snail genus Solatopupa (Pulmonata) in the peri-Tyrrhenian area.

The land snail genus Solatopupa consists of six species and has a peri-Tyrrhenian distribution; most of the species have a very narrow range and all of them except one (Solatopupa cianensis, which inhabits porphyritic rocks) are strictly bound to calcareous substrates. One species (Solatopupa guidoni) is limited to Sardinia, Corsica, and Elba Island. Because the potential for dispersal of these snails is low, the insular range of this species has been traditionally related to the Oligocenic detachment of the Sardinia-Corsica microplate from the Iberian plate and its subsequent rotation towards the Italian peninsula. In this study, we used sequences of three mitochondrial and one nuclear gene to reconstruct the evolutionary history of the genus. Our phylogenetic results are consistent with the genetic relationships found using allozymes, but contrast with the phylogenetic hypotheses based on karyology and morphology. Molecular clock estimates indicate that the main cladogenetic events in the genus occurred between the middle Miocene and the middle-late Pliocene. Patterns of phylogenetic relationships and geological considerations suggest that the cladogenesis of the genus can be explained by vicariant (tectonic) processes. Our datings do not support a causal relation between the split of S. guidoni from its continental sister taxon and the initial phases of the detachment of the Corsica-Sardinia microplate from the mainland. On the contrary, time estimates coincide with the very last phase of detachment of the microplate (from 5 to 3 Myrs ago). Overall, our molecular clock estimates are in good agreement with the latest geological views on the tectonic evolution of the peri-Tyrrhenian area.

The seminal receptacle and sperm storage in Cochlostoma montanum (Issel) (Gastropoda: Prosobranchia).

Females of Cochlostoma montanum (Prosobranchia, Cochlostomatidae) have a seminal receptacle which is not a separate diverticulum of the oviduct. The seminal receptacle derives from a differentiated portion of the renal oviduct which has an inner wall composed of only one layer of cells. These cells are of two different types, both actively involved in secretory activity. One type is represented by goblet cells filled with large vesicles containing an electron-dense, homogeneous, and partially paracrystalline material. This material is expelled into the lumen through macro-apocrine or holocrine types of secretion. The other type is represented by ciliated cells rich in small vesicles containing granular material. Probably neither kind of secretion has a nutritive function; rather they serve as matrix for spermatozoa that immobilize them and prevent their expulsion from the receptacular portion of the oviduct. Spermatozoa are inserted in invaginations of the apex of both these epithelial cells. The sperm plasma membrane covering the acrosome forms long digitations which expand toward the corresponding invaginations of the receptaculum cells. This type of adhesion is a novelty for Mollusca and resembles that in seminal receptacles of some Annelida.