The systematics and evolution of the Sri Lankan rainforest land snail Corilla: New insights from RADseq-based phylogenetics.

The stylommatophoran land-snail genus Corilla is endemic to Sri Lanka and India's Western Ghats. On the basis of habitat distribution and shell morphology, the 10 extant Sri Lankan species fall into two distinct groups, lowland and montane. Here, we use phylogenetic analyses of restriction-site-associated DNA sequencing (RADseq) data and ancestral-state reconstructions of habitat association and shell morphology to clarify the systematics and evolution of Sri Lankan Corilla. Our dataset consists of 9 species of Corilla. Phylogenetic analyses were based on 88 assemblies (9,604-4,132,850 bp) generated by the RADseq assembler ipyrad, using four parameter combinations and different levels of missing data. Trees were inferred using a maximum likelihood (ML) approach. Ancestral states were reconstructed using maximum parsimony (MP) and ML approaches, with 1 binary state character analysed for habitat association (lowland vs montane) and 6 binary state characters analysed for shell morphology (shape, colour, lip width, length of upper palatal folds, orientation of upper palatal folds and collabral sculpture). Over a wide range of missing data (40-87 % missing individuals per locus) and assembly sizes (62,279-4,132,850 bp), nearly all trees conformed to one of two topologies (A and B), most relationships were strongly supported and total branch support approached the maximal value. Apart from the position of Corilla odontophora 'south', topologies A and B showed similar, well-resolved relationships at and above the species level. Our study agrees with the shell-based taxonomy of C. adamsi, C. beddomeae, C. carabinata, C. colletti and C. humberti (all maximally supported as monophyletic species). It shows that C. erronea and C. fryae constitute a single relatively widespread species (for which the valid name is C. erronea) and that the names C. gudei and C. odontophora each apply to at least two distinct, yet conchologically-cryptic species. The MP and ML ancestral-state reconstructions yielded broadly similar results and provide firm evidence that diversification in Sri Lankan Corilla has involved evolutionary convergence in the shell morphology of lowland lineages, with a pale shell and wide lip having evolved on at least two separate occasions (in C. carabinata and C. colletti) from montane ancestors having a dark, narrow-lipped shell.

Species limits, interspecific hybridization and phylogeny in the cryptic land snail complex Pyramidula: The power of RADseq data.

Restriction site-associated DNA sequencing (RADseq) was used to jointly assess phylogenetic relationships, interspecific hybridization and species delimitation in the cryptic, non-model land snail complex Pyramidula. A robust phylogeny was inferred using a matrix of concatenated sequences of almost 1,500,000bp long, containing >97,000 polymorphic sites. Maximum likelihood analyses fully resolved the phylogenetic relationships among species and drastically improved phylogenetic trees obtained from mtDNA and nDNA gene trees (COI, 16S rRNA, 5.8S rRNA, ITS2 and 28S rRNA sequence data). The best species delimitation scenario was selected on the basis of 875 unlinked single nucleotide polymorphisms, showing that nine Pyramidula species should be distinguished in Europe. Applying D-statistics provided no or weak evidence of interspecific hybridization among Pyramidula, except for some evidence of gene flow between two species.

Molecular identification of novel intermediate host species of Angiostrongylus vasorum in Greater London.

Angiostrongylus vasorum is a parasitic nematode that can cause serious and potentially fatal disease in dogs and other canids. The aim of this study was to determine the intermediate slug species infected in nature by sampling sites in Greater London and Hertfordshire located within a known hyperendemic region. Overall, A. vasorum larvae were recovered from 6/381 slugs (1.6%) by tissue digestion, and their identity was confirmed by PCR. Infected slugs originated from three different sites in the Greater London area: one in Waltham Forest and two in Bromley. Slugs parasitised by A. vasorum were identified by a combination of external morphological characteristics and molecular techniques and belonged to three different families: the Arionidae, the Milacidae and the Limacidae. This includes two new host records for the parasite: Arion distinctus and Tandonia sowerbyi. This is the first record of A. vasorum in the family Milacidae, indicating that the parasite has a broader intermediate host range than previously recognised.

Mixed breeding system in the hermaphroditic land slug Arion intermedius (Stylommatophora, Arionidae).

Theory suggests that hermaphroditic plants and animals should be either entirely outcrossing or entirely selfing. As such, very few hermaphroditic plants and basommatophoran snails have a mixed breeding system. However, reliable estimates of selfing rates are lacking for most hermaphroditic animals. This partly prevents to delineate the relative contributions of the selective factors that determine selfing and outcrossing rates in hermaphroditic animal taxa. Here, we studied the population genetic structure of, and breeding system in, 11 populations of the hermaphroditic land slug Arion intermedius using five polymorphic microsatellite loci. Moreover, genotype frequencies deviated significantly from Hardy-Weinberg equilibrium expectations for most of the loci in all populations suggesting some level of selfing. Estimates of the selfing level s, suggest moderate levels of outcrossing (mean s based on FIS = 0.84; mean s based on the two-locus heterozygosity disequilibrium = 0.20, or with a ML approach = 0.22). Our study therefore suggests that A. intermedius has a mixed breeding system. A re-analysis of allozyme data from another arionid slug ( subgenus Carinarion) indicates that mixed breeding may be more common in arionid slugs than hitherto was assumed. These results seem therefore at variance with current theoretical and empirical predictions and opens perspectives for the study on the evolutionary factors driving mixed breeding systems in animals.

Rapid large-scale evolutionary divergence in morphology and performance associated with exploitation of a different dietary resource.

Although rapid adaptive changes in morphology on ecological time scales are now well documented in natural populations, the effects of such changes on whole-organism performance capacity and the consequences on ecological dynamics at the population level are often unclear. Here we show how lizards have rapidly evolved differences in head morphology, bite strength, and digestive tract structure after experimental introduction into a novel environment. Despite the short time scale ( approximately 36 years) since this introduction, these changes in morphology and performance parallel those typically documented among species and even families of lizards in both the type and extent of their specialization. Moreover, these changes have occurred side-by-side with dramatic changes in population density and social structure, providing a compelling example of how the invasion of a novel habitat can evolutionarily drive multiple aspects of the phenotype.

Morphological and molecular characterization of Klossnema viguerasi n. sp. (Nematoda: Oxyuridomorpha: Hystrignathidae) from a Cuban passalid beetle (Coleoptera: Passalidae), first record of the genus for Cuba.

Klossnema viguerasi n. sp. (Nematoda: Oxyuridomorpha: Hystrignathidae) is described from the passalid beetle Antillanax pertyi (Kaup, 1869), endemic to Cuba. The females of K. viguerasi n. sp. are morphologically similar but slightly longer than K. repentina Cordeiro Artigas, 1983 (1.143 mm vs. 1.000 mm). Both species differ in that K. viguerasi n. sp. has a longer procorpus (139 µm vs. 110 µm), isthmus (39 µm vs. 24 µm), and tail length (28 µm vs. 21 µm). The distance from the vulva to the anterior end is also longer in the new species (0.748 mm vs. 0.650 mm). The males of K. viguerasi n. sp. are larger than K. repentina (0.980 mm vs. 0.800 mm), but their isthmus is shorter (38 µm vs. 48 µm). New features of the cephalic end of both sexes, and copulatory papillae pattern of the males were observed by SEM and the generic diagnosis is emended in order to include such features. The phylogeny of K. viguerasi n. sp. is inferred by the analysis of the D2-D3 domains of the 28S rDNA and the 18S rDNA. This constitutes the first record of the genus Klossnema for the Cuban archipelago and the West Indies.

Intermediate gastropod hosts of major feline cardiopulmonary nematodes in an area of wildcat and domestic cat sympatry in Greece.

BACKGROUND: The metastrongyloid nematodes Aelurostrongylus abstrusus, Troglostrongylus brevior and Angiostrongylus chabaudi are cardiopulmonary parasites affecting domestic cats (Felis catus) and wildcats (Felis silvestris). Although knowledge on these nematodes has been improved in the past years, gaps in our knowledge of their distribution and role of gastropods as intermediate hosts in Europe still exist. This study reports on the presence of these nematodes and their intermediate hosts in an area in Greece where domestic cats and wildcats occur in sympatry. METHODS: Terrestrial gastropods were collected in the field and identified morphologically and by mitochondrial DNA-sequence analysis. Metastrongyloid larvae were detected by artificial digestion, morphologically identified to the species and stage level and their identity was molecularly confirmed. RESULTS: Aelurostrongylus abstrusus was found in the snails Massylaea vermiculata and Helix lucorum, T. brevior in the slug Tandonia sp., and A. chabaudi in the slug Limax sp. and the snails H. lucorum and M. vermiculata. CONCLUSIONS: To the best of our knowledge this study provides the first reports of (i) terrestrial gastropods being naturally infected with A. chabaudi, (ii) T. brevior naturally infecting terrestrial gastropods in Europe, and (iii) A. abstrusus naturally infecting terrestrial gastropods in Greece. Furthermore, the present study describes for the first time developmental stages of A. chabaudi and T. brevior in naturally infected gastropods. The biological characteristics of various intermediate gastropod hosts that could influence the distribution and expansion of feline cardiopulmonary nematodes are discussed, along with epizootiological implications and perspectives.

Morphological and molecular characterization of Lepidonema magnum Morffe García, 2010 (Nematoda: Oxyuridomorpha: Hystrignathidae) from Passalus interstitialis Eschscholtz, 1829 (Coleoptera: Passalidae) from Cuba and new locality records for the species.

Lepidonema magnum Morffe García, 2010 (Nematoda: Oxyuridomorpha: Hystrignathidae) is redescribed and illustrated with the aid of SEM. New features of the cephalic end and genital tract of the females were observed. New locality records are given. The phylogenetic position of the species is inferred on the basis of the D2-D3 segment of the 28S LSU rDNA and 18S SSU rDNA. L. magnum forms a monophyletic clade formed by other hystrignathids: Coynema poeyi (Coy, García Álvarez, 1993), two species of Longior Travassos Kloss, 1958 and two Hystrignathus Leidy, 1850.

Morphological examination and phylogenetic analysis clarify the taxonomic status of Cuban Longior Travassos Kloss, 1958 (Nematoda: Thelastomatoidea: Hystrignathidae).

The status of Longior alius García Coy, 1994 (Nematoda: Thelastomatoidea: Hystrignathidae) is discussed on the basis of new material obtained from its type host Antillanax pertyi (Kaup, 1869) (Coleoptera: Passalidae) from Limonar, Guantánamo province, Cuba, near the type locality. Based on morphological and molecular studies L. alius is considered synonymous with L. longior Morffe García, 2011. L. longior and L. similis Morffe, García Ventosa, 2009 are redescribed and illustrated with the aid of SEM. New locality records for both species are given. The conspecificity of females and males of both species is supported by comparison of the D2-D3 segment of the 28S LSU rDNA. The inter-specific differences and phylogenetic position of L. longior and L. similis are discussed.

Morphological and molecular characterization of Coynema poeyi (Coy, García Álvarez, 1993) (Oxyuridomorpha: Hystrignathidae) from Antillanax pertyi (Kaup, 1869) (Coleoptera: Passalidae) from Cuba and new locality records for the species.

Coynema poeyi (Coy, García Álvarez, 1993) (Oxyuridomorpha: Hystrignathidae) is redescribed and illustrated with the aid of SEM. New features of the cephalic end of both sexes and copulatory papillae pattern of the males were observed and the generic diagnosis is emended in order to include such features. New locality records are given. The phylogenetic position of the species is inferred on the basis of the D2-D3 segment of the LSU rDNA. C. poeyi is located basal in a monophyletic clade formed by other hystrignathids: two species of Longior Travassos Kloss, 1958 and Hystrignathus sp.

DNA barcodes and phylogenetic affinities of the terrestrial slugs Arion gilvus and A. ponsi (Gastropoda, Pulmonata, Arionidae).

The Iberian Peninsula is a region with a high endemicity of species of the terrestrial slug subgenus Mesarion. Many of these species have been described mainly on subtle differences in their proximal genitalia. It therefore remains to be investigated 1) whether these locally diverged taxa also represent different species under a phylogenetic species concept as has been shown for other Mesarion species outside the Iberian Peninsula, and 2) how these taxa are phylogenetically related. Here, we analysed DNA sequence data of two mitochondrial (COI and 16S) genes, and of the nuclear ITS1 region, to explore the phylogenetic affinities of two of these endemic taxa, viz. Arion gilvus Torres Mínguez, 1925 and A. ponsi Quintana Cardona, 2007. We also evaluated the use of these DNA sequence data as DNA barcodes for both species. Our results showed that ITS did not allow to differentiate among most of the Mesarion molecular operational taxonomic units (MOTUs) / morphospecies in Mesarion. Yet, the overall mean p-distance among the Mesarion MOTUs / morphospecies for both mtDNA fragments (16.7% for COI, 13% for 16S) was comparable to that between A. ponsi and its closest relative A. molinae (COI: 14.2%; 16S: 16.2%) and to that between A. gilvus and its closest relative A. urbiae (COI: 14.4%; 16S: 13.4%). Hence, with respect to mtDNA divergence, both A. ponsi and A. gilvus, behave as other Mesarion species or putative species-level MOTUs and thus are confirmed as distinct 'species'.

First report of the exotic blue land planarian, Caenoplana coerulea (Platyhelminthes, Geoplanidae), on Menorca (Balearic Islands, Spain).

In April 2009 two specimens of a terrestrial flatworm were collected from under a rock in an orchard at Ciutadella de Menorca on the easternmost Balearic island of Menorca (Spain). Their external morphology suggested that both specimens belonged to the invasive blue planarian Caenoplana coerulea, a species which is native to eastern Australia. Sequence data of a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) and of the entire 18S ribosomal RNA confirm its identification. This is one of the first records of the species in Europe where it has only been found in one locality in the United Kingdom, France and NE Spain.