NEWS OF PSEUDAMNICOLA ( CORROSELLA ) OF SPAIN AND FRANCE ( MOLLUSCA : GASTROPODA : TRUNCATELLOIDEA )

Pseudamnicola (Corrosella) Boeters, 1970 from Spain and France is reviewed: delimitation of the subgenus against the nominotypical subgenus is supported with geographical, anatomical and conchological data. Four new species are described, three from Spain: P. (C.) collingi n. sp., P. (C.) valladolensis n. sp. with two subspecies P. (C.) v. valladolensis n. ssp. and P. (C.) v. kahbei n. ssp., and P. (C.) tajoensis n. sp., and one from France: P. (C.) tejedoi n. sp. Another two species: P. (C.) hinzi Boeters, 1986 and P. (C.) navasiana (Fagot, 1907), are revised. DNA barcodes (COI) were analysed for some of the new species. Key worDs: Pseudamnicola, Corrosella, subgenus delimitation, new species, COI, mitochondrial DNA

The genus comprises two subgenera, the nominotypical subgenus and Corrosella Boeters, 1970.Whereas species of the nominotypical subgenus have been reported from the whole distribution area of the genus, representatives of Corrosella have been recorded only from France (Boeters 2000, GirarDi et al. 2009) and Spain (Boeters 1988) yet.
Recently DeliCaDo et al. (2012) have distinguished Corrosella from the nominotypical subgenus, i.e.Pseudamnicola s. str., based on five synapomorphies.Corrosella is characterised by a more slender shell and a more pigmented and tapered penis, compared to Pseudamnicola s. str., whereas the latter has a longer seminal receptacle and a longer bursa duct.Furthermore, in Corrosella the bursa copulatrix varies from pyriform to cylindrical, while in Pseudamnicola s. str. it is ovoid to pyriform (HersHler & PonDer 1998).
DeliCaDo et al. (2012) and DeliCaDo & ramos (2012) have redefined the criteria for distinguishing among the species of Corrosella.In the light of their work it is obvious that P. (Corrosella) hinzi as originally described (Boeters 1986) covers two separate species.Thus, P. (C.) hinzi is redefined and a new species, P. (C.) collingi n. sp., is described in this paper.
With one exception, members of Corrosella inhabit drainage systems supplying the Mediterranean Sea and the adjacent Atlantic Golfo de Cádiz.The first and single report of a representative of Corrosella inhabiting a system draining the Sistema Central into the Portuguese Atlantic, has been provided by soler et al. (2006).This paper confirms and elucidates it by the description of P. (C.) valladolensis n. sp. and P. (C.) tajoensis n. sp.
A new species of Corrosella, P. (C.) tejedoi n. sp., is described from the area between Zaragoza and the French Côte-d'Azur, where no member of Corrosella has been found previously.
DNA sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene fragment are frequently used for species identification on molecular level (DNA barcoding; introduced by HeBert et al. 2003).In this study, we analysed COI DNA barcodes, in order to assess the discrimination of the studied taxa at the molecular level.

MATERIAL AND METHODS
Whorls were counted according to GittenBerGer et al. (1970).The shell measurements are presented as minimum-mean-maximum values.The colours of shells and opercula apply to samples kept in ethanol 75% by weight; in dried state, the colour of shells and opercula changes from white to corneous or from corneous to brownish.Anatomical examination followed Boeters (1999).
Drawings of shells and animals and parts thereof were done with ZEISS 45 degree Drawing Prism.Except for Tables 2-3 and Fig. 71, the shell length and diameter were measured with a 5 mm measure plate (0.05 mm grading) at 25× magnification; the measurements were rounded to the nearest 0.05 mm.For the data in Tables 2-3 and Fig. 71 a micrometer was used (reticular ocular 10 : 100).Photographs of shells were taken with a stereomicroscope ZEISS 2000 C, colour photographs with a digital colour camera SSC-DC SONY with monitor exit TRINITON COLOR VIDEO SONY, to achieve depth of focus.The photographs were touched up with PAINT SHOP PRO (version 9).
The distribution map (Fig. 1, see also: Appendix 1) is based on MGRS (Military Grid Reference System).
For molecular analysis we used 32 COI sequences published by DeliCaDo et al. ( 2013) and deposited at GenBank (http://www.ncbi.nlm.nih.gov)under accession numbers JX081854-JX081885.All of the 32 sequences were originally assigned to P. navasiana (DeliCaDo et al. 2013).The sequences were aligned using Muscle algorithm (eDGar et al. 2004) implemented in MEGA version 6.06 (tamura et al. 2011).Intra-and interspecific genetic distances were calculated in MEGA using the Kimura two-parameter (K2P) model (Kimura 1980).The difference between the maximum intraspecific distance and the smallest distance to the nearest neighbour (NN) is known as the "barcode gap" and quantifies the distinctness of species at the DNA barcode locus (meyer & Paulay 2005).Barcode gaps were calculated for each species.A neighbour-joining (NJ) topology (saitou & nei 1987) was built based on K2P pairwise genetic distances together with bootstrap analysis of 1,000 replicates (Felsenstein 1985).
The material is kept in the following collections: BOE -Collection Hans Boeters (München); HG -Collection Henri Callot-GirarDi (Montfavet); SMF -Forschungsinstitut und Natur-Museum Senckenberg (Frankfurt am Main).Animal : On visceral sac whorls light to dark brown spots.Mantle except for its white border pigmented brownish, only eyes, tentacles and head between tentacles pigmented black.Foot seen through operculum with three parallel colour fields running to the tip of operculum, from left to right: brown, white (under nucleus) and brownish.Separated operculum uniformly corneous to brownish.Gill (Fig. 40) with 17 filaments (n = 1♂).
Female genitalia : The proximal, first section of renal oviduct is pigmented black and coiled up to the distal, final section which is straight, only very slightly pigmented and entered by two ducts: the receptaculum duct, and the bursal duct; duct of the small drop-to sac-like receptaculum (rs1) very short; bursa copulatrix long, U-like bent; bursal duct thick, gradually passing into the bursa in front of its bend.The receptaculum reaches the bursa bottom  or downward; peristome smooth.Periostracum more or less eroded, the degree of erosion increases from the last to the first whorl, sometimes to such a degree that the eroded areas are covered by calcareous structures.
Penis : Slender, tapered from base to tip; the penial duct undulates slightly along the penis periphery and is flanked by a weakly blackish pigmented zone of the penis core; the duct ends nearly at the tip of penial body (n = 1 each from Covanera and Tubilla del Agua).
Female genitalia : The proximal, first section of renal oviduct is pigmented black and coiled up to the distal, final section which is straight, only very slightly pigmented and entered by two ducts, the receptaculum duct and the bursal duct; duct of the small drop-to sac-like receptaculum (rs1) very short; bursa copulatrix long, U-like bent; bursal duct thick, gradually passing into the bursa in front of its bend.The receptaculum reaches the bursa bottom (n = 2 from Covanera).Differentiating characters: 1. Contrary to P. (Corrosella) hinzi the mantle is not pigmented brownish, but black except for its white border.The penial duct is flanked by a slightly blackish pigmented zone of its core.Furthermore, the penis is tapered from base to tip and not finger-like.81) and showed the genetic distance to the nearest neighbouring species of 1.8% (Table 5).This new species inhabits the drainage system of the Rio Duero, with 2 subspecies.Animal : Head between tentacles, eyes, tentacles and mantle except for its white border pigmented black.Visceral sac whorls with lighter and darker pigmented spots.Foot seen through operculum with three parallel colour fields running to the tip of operculum, from left to right: black, brownish (under nucleus) and black.Separated operculum uniformly light brownish except for the slightly more intensively brownish coloured nucleus.Gill (Fig. 42) with 14 filaments (n = 1♀).Penis (Figs [14][15][16][17]: Tapered from base to tip, in its resting position with weak transverse folds; penis bent, so that the left, inner periphery of the bend looks slightly wavy; penial duct meandering towards the penis tip and accompanied by a slightly blackish zone of the penis core; remarkably, three out of the four dissected males showed a different shape each (n = 4).up to the distal, final section which is straight, only very slightly pigmented and entered by two ducts, the receptaculum duct and the bursal duct; duct of the small drop-to sac-like receptaculum (rs1) extremely short; bursa copulatrix not oriented parallel to the intestine, but embedded in the albumen gland; the bursa is slim and long, U-like bent, both arms oriented parallel to each other, but at a distance from each other; bursal duct gradually passing into the bursa in front of its bend.The receptaculum does not touch the bursa bottom (n = 3    The proximal, first section of renal oviduct is unpigmented or at most very weakly pigmented black and coiled up to the distal, final section which is straight, at most very slightly pigmented and entered by two ducts, the receptaculum duct and the bursal duct; duct of the small drop-to sac-like receptaculum (rs1) extremely short; bursa copulatrix long and voluminous, U-like bent; bursal duct thicker than the renal oviduct, slightly separated from the bursa or gradually passing into the bursa in front of its bend.
The receptaculum reaches the bursa bottom (n = 2).1998: 372), and supplying the Atlantic Ocean, not the Mediterranean Sea. 2. The NJ analysis (Fig. 81) revealed one well supported monophyletic cluster including sequences from Peralejos (Per), Ontígola (Mar) and Borox (Box) published by DeliCaDo et al. (2013: 390-391).The distance between the "Mar" population and the type locality of P. (C.) tajoensis n. sp. is probably less than 3 km.Accordingly, the sequences can be assigned to P. (C.) tajoensis n. sp. and show a genetic distance to the nearest neighbouring species of 1.2 % (Table 5).

Specific characters of the penis are additionally
commented on in the Discussion.4).Distribution and habitat: Known from the type locality only, the Source de la Rivière d'Err (Font de la Ribera d'Err) in the French Pyrenees (Pyrénées- Orientales), sympatric with Bythinella sp.(BOE 3157).The river Err is a tributary of the Segre and belongs to the Ebro drainage system.Remarks: This species is the second representative of Corrosella in France, but it inhabits a drainage system which currently drains the waters of the Pyrenees to the river Ebro in Spain.Since its description is based on conchological and distributional characters only, further investigations, especially of anatomical and genetic characters, are necessary to establish the species identity and distinctness.80, Table 1)

Pseudamnicola (Corrosella
1907 Amnicola navasiana Fagot (FaGot 1907: 158), type locality: "Buelbuente [sic]" 1988 Pseudamnicola (Corrosella) navasiana (Boeters 1988: 203, figs 65, 71, 87, pl. 2 fig. 20) Material examined: Spain, Zaragoza, Fonnueva spring about 2 km west of Bulbuente; Hinz leg.IV.1984 [label: "Zara 1"]; BOE 1239/animals + BOE 1654/shells.Shell (Figs 72,74,76,(78)(79)(80): Ovate-conical with strongly vaulted whorls; body whorl often with a slight shoulder between suture and periphery; initial whorls more or less eroded; apex can project like a corksrew or be completely lost and replaced by whitish deposits; shell surface gives a spotted impression, because the whitish eroded areas alternate with periostracum fragments; areas of intact periostracum and especielly fractures of broken shells give an opaque to milky impression resembling shells of Mercuria; shells of mature animals may still have about 4 whorls; aperture inclined ovate; palatal and parietal border forming a rounded angle of slightly less than 90°; parietal border broadened, but generally forming a slit with the shell wall, only occasionally fused with it over a short distance; umbilical border often remarkably broadened without covering the umbilicus; umbilicus, however, often closed by whitish deposits; peristome sharp.Shell measurements (Table 1): Height (h) 3.05-3.34-3.45mm, diameter (d) 1.85-2.08-2.25 mm (n = 5 each), h : d ratio 1.61.Animal (Figs 73,77): Visceral sac whorls with light to intensively darkish spots.Mantle except for its white border, tentacles with eyes and head between tentacles pigmented black.Foot seen through operculum with three parallel colour fields running to the tip of operculum, from left to right: blackish, white (under nucleus) and blackish.Separated operculum uniformly corneous to brownish.Gill with 15 to 16 filaments (n = 2♀♀).Penis (Fig. 73): Broad tapering penis base bent like a flat U towards the mantle skirt, followed by a slowly tapering slender final section; the penial duct runs straight along the penis periphery, flanked by a slightly blackish pigmented zone of the penis core; behind its bend and opposite to the penial duct, the penis base looks like a saw blade due to unpigmented transverse folds; the penial duct ends nearly at the tip of penial body and forms with it a rounded penis tip (n = 3).Female genitalia (Fig. 75): The proximal first section of renal oviduct is pigmented black and coiled up to the distal, final section which is straight, only very slightly pigmented and entered by two ducts, the receptaculum duct and the bursal duct; duct of the small drop-to sac-like receptaculum (rs1) very short; bursa copulatrix long, U-like bent; bursal duct and bursa are clearly separated from each other, and the receptaculum does not reach the bursa bottom (n = 2).Differentiating characters are given under P. Remarks: Already the original description characterises the shell as opaque, but muddy greyish ["opaca… sordide grisea"], a character which resembles milky shells of Mercuria species.The DNA sequences of the P. (C.) navasiana populations formed a monophyletic cluster (Fig. 81) indicating genetic separation from the other species.The genetic distance to the nearest neighbouring species was found to be 1.2% (Table 5).Hinz' vague indication concerning specimens collected in April 1984 "spring about 2 km west of Bulbuente" could be identified as Fonnueva spring; cf.DeliCaDo et al. (2013: 390) Altaba, 2007(Boeters 1981, altaBa 2007, Glöer & zettler 2007).For the rediscovery of P. (Pseudamnicola) spirata (= subproducta) in the region of its type locality, "aux environs de Banolas (Catalogne)", see Boeters (1988: 199) Boeters (1970Boeters ( , 1984) ) and DeliCaDo et al. ( 2012) independently of each other showed that characters of receptaculum and bursa could be useful for differentiation of species.The receptaculum can be positioned at a distance from the bursa or touch or lean on it (luisi : Boeters 1984: 10, fig. 2, Boeters 1988: 204, fig. 88, DeliCaDo et al. 2012: 35, fig. 4H;falkneri: Boeters 1970: 68, fig. 8, Boeters 1988: 204, figs 90-91, DeliCaDo et al. 2012: 39, figs 7H-J).Furthermore, the duct of the bursa can be clearly separated from the voluminous bursa as in P. (C.) falkneri, or the duct can gradually pass into the bursa as in P. (C.) hinzi (Boeters 1986: 126, fig. 2, Boeters 1988: 204, fig. 85).17D).Starting from the discovery and anatomical examination of P. (Corrosella) tajoensis n. sp., this character, needs, however, clarification.In P. (C.) tajoensis n. sp. the penial duct is not simply embedded in the penis body but flanks the body on its external side.The penis tip protrudes beyond the opening of the duct in such a manner that the end of the duct forms a shoulder at the periphery of the penis in front of its tip.A re-examination of other species of Corrosella now revealed that the described structure applies also to other species, such as P. (C.) navasiana, but is not necessarily as pronounced as in P. (C.) tajoensis n. sp.This suggests that similar characters might be present in nearly all species of Corrosella.locality which corresponds to this indication is Spéracèdes, not more than 5 km west of the centre of Grasse.A comparison of shells from this locality with shells from three other localities about 40, 75 and 85 km west of Grasse is given in Table 6.

PENIS
Shells from Draguignan, Pontevès and Seillons are smaller than those from Spéracèdes and more elongated which is reflected by the h:d ratio of 1.55, 1.67 (or even 1.78) and 1.73 instead of 1.47.Because of these differences which make shells from Pontevès, for example, more similar to shells of P. (C.) tejedoi n. sp.(Fig. 71), a critical reconsideration of the identity of P. (C.) astieri seems necessary.
In this context it is noteworthy that DuPuy (1851) described his species as having its shell "3-4 mill."high, which corresponds best to the measurements given for Spéracèdes (Table 6) and the only syntype which has yet come to our knowledge.The shell height of the syntype is 3.05 mm.It will be described in the context of the intended critical reconsideration.

Fig. 1 .
Fig. 1.Distribution of species of subgenera Pseudamnicola (asterisks) and Corrosella (circles) in Spain and the mainland of France.For details see Appendix 1.The distribution areas of the two genera roughly separated with a solid line

2 .
In P. (Corrosella) navasiana the suture is deeper than in P. (Corrosella) collingi n. sp.Besides, in P. (C.) navasiana the body whorl often shows a slight shoulder between the suture and the periphery, whereas in P. (C.) collingi n. sp.such a shoulder is absent.In P. (Corrosella) navasiana the bursal duct and bursa are clearly separated from each other, whereas in P. (Corrosella) collingi n. sp. the bursal duct passes gradually into the bursa in front of its bend.In P. (C.) navasiana the receptaculum is located at a distance from the bursa, whereas in P. (C.) collingi n. sp. it touches the bursa.In P. (C.) collingi n. sp. the penial duct undulates slightly, whereas in P. (C.) navasiana it runs straight down towards the penis tip. 3.In P. (Corrosella) tajoensis n. sp. the penis is unpigmented, whereas in P. (Corrosella) collingi n. sp. the penial duct is flanked by a slightly blackish pigmented zone of the penis core.Furthermore, in P. (C.) tajoensis n. sp. the penial duct runs at the periphery of the penis body straigth to the penis tip, while in P. (C.) collingi n. sp. the penial duct undulates slightly from the base of the penis up to its tip.
Female genitalia (Figs 30-33): The proximal, first section of renal oviduct pigmented black and coiled

2 .
Being 3.22 mm high, the shells of P. (Corrosella) v. valladolensis n. ssp.are on average slightly larger than those of P. (Corrosella) tajoensis n. sp.(2.90 mm).Compared to P. (Corrosella) v. kahbei n. ssp., the vault of the whorls below the suture is well rounded without any indication of even a weak shoulder.In P. (C.) v. valladolensis n. ssp.and in P. (C.) v. kahbei n. ssp.there is no shoulder at the end section of penis, immediately in front of its tip.3. Contrary to the new species, in P. (Corrosella) navasiana the suture is deeper and the body whorl
Reference is especially made to the figures given by DeliCaDo et al. (2012) and DeliCaDo & ramos (2012) for P. (C.) bareai, P. (C.) marisolae and P. (C.) hauffei.These figures show a slight step or shoulder in front of the penis tip despite the fact that according to the figures the duct runs directly up to the penis tip (DeliCaDo et al. 2012: 54, fig.14G, 59, fig.17D, DeliCaDo & ramos 2012: 72, fig.7D).The flanking of the penis body by the penial duct might reflect the origin of the closed duct from an open gutter as is known, for example, in the Aciculidae.There is, however, a restricted number of species of Corrosella with a slightly undulating, wavy or meandering instead of straight course of the penial duct towards its opening, for example P. (C.) manueli and P. (C.) marisolae (DeliCaDo et al. 2012: 49, 59, fig.17D and 60) and P. (C.) collingi n. sp.It would be interesting to clarify whether this structure has simply to do with changes of the penis from its resting to active state or whether evolutionary aspects should be considered.PSEUDAMNICOLA (CORROSELLA) TEJEDOI N. SP.In the southern foreland of the Pyrenees, species of Corrosella are known only from its southwestern part and only from the drainage systems of southern tributaries of the Ebro River (DeliCaDo et al. 2013: 389, fig.1).They are completely unknown from the Pyrenees themselves and their northern foreland.The only representative of the subgenus lives far to the northeast of the Pyrenees, in Southeastern France.DeliCaDo et al. (2013: 395) interpreted the Pyrenees as a geographical barrier.They argued that in the northern part of the Iberian Peninsula, up to Southern France, "some populations [of P. (Corrosella) species] were isolated probably due to the last uprising of the Pyrenees during the upper Miocene.(…) This geological barrier may have led to the split between P. (C.) astieri and the group formed by P. (C.) navasiana and P. (C.) hauffei."Thus, the discovery of a representative of Corosella in the drainage systems of a northern tributary of the Ebro River and directly in the Pyrenees was unexpected.THE TYPE LOCALITY OF PSEUDAMNICOLA (CORROSELLA) ASTIERI 1 DuPuy (1851) reported "les environs de Grasse" as the type locality for P. (C.) astieri.The only known 1 The species was originally described by DuPuy (1851) as Hydrobia astierii with its name dedicated to M. Astier.The species name was amended by DeliCaDo & ramos (2012) according to the current ICZN rules.See also Bou-CHet P. (2014).Pseudamnicola astieri Dupuy, 1851.Accessed through World Register of Marine Species at http://www.marinespecies.org/aphia.php?p=taxdetails&id=716159 on 2014-11-10 (editor's remarks).
(C.) navasiana the receptaculum is located at a distance from the bursa, whereas in P. (C.) tajoensis n. sp. it touches the bursa.In P. (C.) navasiana the penial duct ends nearly at the tip of the penial body and forms with it a rounded tip, whereas in P. (C.) tajoensis n. sp. the duct ends in front of the tip of the penial body, forming a small shoulder immediately in front of the tip, which protrudes like a wedge.4. Characters differentiating P. (Corrosella) tajoensis n. sp.from P. (Corrosella) collingi n. sp. are given under P. (Corrosella) collingi n. sp.Habitat and distribution:

Table 1 )
. Furthermore, in frontal view the body whorl of P. (C.) hauffei is flattened (DeliCaDo & ramos 2012: 68, figs 5A-C), whereas the whorls of P. (C.) tejedoi n. sp. are well vaulted.4. Shells of P. (C.) astieri (Figs 51, 68-70, Table 3, Appendix 2B) are less elongatedly conical (Table 1); in P. (C.) navasiana the parietal border of the aperture does not always touch the shell wall and the umbilicus is more than slit-like open (cf.Boeters 1970: 69 fig.9; DeliCaDo & ramos 2012: 62, fig.2A). 5. P. (C.) tejedoi n. sp.differs from all other known species of Corosella no only in the conchological characters, but also in being geographically sep- arated by a distance of certainly more than 300 km from P. (C.) navasiana and P. (C.) hinzi, more than 350 km from P. (C.) hauffei (type locality) and more than 400 km from P. (C.) astieri, which live next to the Pyrenees.Besides, the altitude of 2,380 m of the habitat of the new species is extraordinary (Table

Table 5 .
. P. (Pseudamnicola) gasulli was reported from coast-al localities in Almeria and Murcia and from the Baleares.The remaining four Spanish species of the nominotypical subgenus were described from the Baleares only.Their description requires a revision of P. (Pseudamnicola) spirata sensu Boeters from Mallorca and Minorca.In France the only well studied species of Pseudamnicola s. s. lives or lived at the border of the Étang de Berre and at the border Intraspecific variation and barcoding gaps for three species based on K2P genetic distances (details seeAppendix 3) by DeliCaDo et al. (2012: 32, figs 2E-F luisi; 37, figs 5F-G falkneri; 62, figs 18F-G iruritai).These erosion traces look like feeding traces.DeliCaDo