António M. de Frias Martins

Phylogenetic relationships and evolution of pulmonate gastropods (Mollusca): New insights from increased taxon sampling

Phylogenetic relationships among higher clades of pulmonate gastropods are reconstructed based on a data set including one nuclear marker (complete ribosomal 18S) and two mitochondrial markers (partial ribosomal 16S and Cytochrome oxidase I) for a total of 96 species. Sequences for 66 of these species are new to science, with a special emphasis on sampling the Ellobiidae, Onchidiidae, and Veronicellidae. Important results include the monophyly of Systellommatophora (Onchidiidae and Veronicellidae) as well as the monophyly of Ellobiidae (including Trimusculus, Otina, and Smeagol). Relationships within Ellobiidae, Onchidiidae, and Veronicellidae are evaluated here for the first time using molecular data. Present results are compared with those from the recent literature, and the current knowledge of phylogenetic relationships among pulmonate gastropods is reviewed: despite many efforts, deep nodes are still uncertain. Identification uncertainties about early fossils of pulmonates are reviewed. Impacts of those phylogenetic and fossil record uncertainties on our understanding of the macro-evolutionary history of pulmonates, especially transitions between aquatic and terrestrial habitats, are discussed.

Ten new complete mitochondrial genomes of pulmonates (Mollusca: Gastropoda) and their impact on phylogenetic relationships.

BackgroundReconstructing the higher relationships of pulmonate gastropods has been difficult. The use of morphology is problematic due to high homoplasy. Molecular studies have suffered from low taxon sampling. Forty-eight complete mitochondrial genomes are available for gastropods, ten of which are pulmonates. Here are presented the new complete mitochondrial genomes of the ten following species of pulmonates: Salinator rhamphidia (Amphiboloidea); Auriculinella bidentata, Myosotella myosotis, Ovatella vulcani, and Pedipes pedipes (Ellobiidae); Peronia peronii (Onchidiidae); Siphonaria gigas (Siphonariidae); Succinea putris (Stylommatophora); Trimusculus reticulatus (Trimusculidae); and Rhopalocaulis grandidieri (Veronicellidae). Also, 94 new pulmonate-specific primers across the entire mitochondrial genome are provided, which were designed for amplifying entire mitochondrial genomes through short reactions and closing gaps after shotgun sequencing.ResultsThe structural features of the 10 new mitochondrial genomes are provided. All genomes share similar gene orders. Phylogenetic analyses were performed including the 10 new genomes and 17 genomes from Genbank (outgroups, opisthobranchs, and other pulmonates). Bayesian Inference and Maximum Likelihood analyses, based on the concatenated amino-acid sequences of the 13 protein-coding genes, produced the same topology. The pulmonates are paraphyletic and basal to the opisthobranchs that are monophyletic at the tip of the tree. Siphonaria, traditionally regarded as a basal pulmonate, is nested within opisthobranchs. Pyramidella, traditionally regarded as a basal (non-euthyneuran) heterobranch, is nested within pulmonates. Several hypotheses are rejected, such as the Systellommatophora, Geophila, and Eupulmonata. The Ellobiidae is polyphyletic, but the false limpet Trimusculus reticulatus is closely related to some ellobiids.ConclusionsDespite recent efforts for increasing the taxon sampling in euthyneuran (opisthobranchs and pulmonates) molecular phylogenies, several of the deeper nodes are still uncertain, because of low support values as well as some incongruence between analyses based on complete mitochondrial genomes and those based on individual genes (18S, 28S, 16S, CO1). Additional complete genomes are needed for pulmonates (especially for Williamia, Otina, and Smeagol), as well as basal heterobranchs closely related to euthyneurans. Increasing the number of markers for gastropod (and more broadly mollusk) phylogenetics also is necessary in order to resolve some of the deeper nodes -although clearly not an easy task. Step by step, however, new relationships are being unveiled, such as the close relationships between the false limpet Trimusculus and ellobiids, the nesting of pyramidelloids within pulmonates, and the close relationships of Siphonaria to sacoglossan opisthobranchs. The additional genomes presented here show that some species share an identical mitochondrial gene order due to convergence.

Using taxonomically unbiased criteria to prioritize resource allocation for oceanic island species conservation

Oceanic islands have been the grand stage of documented extinctions. In view of limited resources, efficient prioritization is crucial to avoid the extinction of taxa. This work lists the top 100 management priority species for the European archipelagos of the Macaronesian region (Azores, Madeira and the Canary Islands), taking into account both their protection priority and their management feasibility. Bryophytes, vascular plants, molluscs, arthropods and vertebrates were scored by species experts following two sets of criteria: (i) protection priority, including ecological value, singularity, public institutions’ management responsibilities and social value; (ii) management feasibility, including threats knowledge and control feasibility, external socio-economical support for management and biological recovery potential. Environmental managers weighted the same criteria according to their management importance. Final species scores were determined by the combination of both species valuation and criteria weighting. Vascular plants dominate the Top 100 list, followed by arthropods and vertebrates. The majority of listed taxa are endemic to one archipelago or even to a single island. The management feasibility criteria did not dictate that all taxa must be eminently endangered, as for most of the species it should be relatively easy to control threats. The main advantages of this process are the independent participation of scientists and conservation managers, the inclusion of criteria on both protection priority and management feasibility and the taxonomically unbiased nature of the process. This study provides a potentially useful biodiversity conservation tool for the Macaronesian archipelagos that could be readily implemented by the respective regional governments in future legislation.

The Azorean Biodiversity Portal: An internet database for regional biodiversity outreach

There is a growing interest in academia to provide biodiversity data to both the scientific community and the public. We present an internet database of the terrestrial lichens, bryophytes, vascular plants, molluscs, arthropods, vertebrates and coastal invertebrates of the Azores archipelago (Portugal, North Atlantic): the Azorean Biodiversity Portal (ABP, http://www.azoresbioportal.angra.uac.pt/). This is a unique resource for fundamental research in systematics, biodiversity, education and conservation management. The ABP was based on a regional species database (ATLANTIS), comprised of grid-based spatial incidence information for c. 5000 species. Most of the data rely on a comprehensive literature survey (dating back to the 19th century) as well as unpublished records from recent field surveys in the Azores. The ABP disseminates the ATLANTIS database to the public, allowing universal, unrestricted access to much of its data. Complementarily, the ABP includes additional information of interest to the general public (e.g. literature on Macaronesian biodiversity) together with images from collections and/or live specimens for many species. In this contribution we explain the implementation of a regional biodiversity database, its architecture, achievements and outcomes, strengths and limitations; we further include a number of suggestions in order to implement similar initiatives.There is a growing interest in academia to provide biodiversity data to both the scientific community and the public. We present an internet database of the terrestrial lichens, bryophytes, vascular plants, molluscs, arthropods, vertebrates and coastal invertebrates of the Azores archipelago (Portugal, North Atlantic): the Azorean Biodiversity Portal (ABP, http://www.azoresbioportal.angra.uac.pt/). This is a unique resource for fundamental research in systematics, biodiversity, education and conservation management. The ABP was based on a regional species database (ATLANTIS), comprised of grid-based spatial incidence information for c. 5000 species. Most of the data rely on a comprehensive literature survey (dating back to the 19th century) as well as unpublished records from recent field surveys in the Azores. The ABP disseminates the ATLANTIS database to the public, allowing universal, unrestricted access to much of its data. Complementarily, the ABP includes additional information of interest to the ge...

Patterns of Diversity of the Rissoidae (Mollusca: Gastropoda) in the Atlantic and the Mediterranean Region

The geographical distribution of the Rissoidae in the Atlantic Ocean and Mediterranean Sea was compiled and is up-to-date until July 2011. All species were classified according to their mode of larval development (planktotrophic and nonplanktotrophic), and bathymetrical zonation (shallow species—those living between the intertidal and 50 m depth, and deep species—those usually living below 50 m depth). 542 species of Rissoidae are presently reported to the Atlantic Ocean and the Mediterranean Sea, belonging to 33 genera. The Mediterranean Sea is the most diverse site, followed by Canary Islands, Caribbean, Portugal, and Cape Verde. The Mediterranean and Cape Verde Islands are the sites with higher numbers of endemic species, with predominance of Alvania spp. in the first site, and of Alvania and Schwartziella at Cape Verde. In spite of the large number of rissoids at Madeira archipelago, a large number of species are shared with Canaries, Selvagens, and the Azores, thus only about 8% are endemic to the Madeira archipelago. Most of the 542-rissoid species that live in the Atlantic and in the Mediterranean are shallow species (323), 110 are considered as deep species, and 23 species are reported in both shallow and deep waters. There is a predominance of nonplanktotrophs in islands, seamounts, and at high and medium latitudes. This pattern is particularly evident in the genera Crisilla, Manzonia, Onoba, Porosalvania, Schwartziella, and Setia. Planktotrophic species are more abundant in the eastern Atlantic and in the Mediterranean Sea. The results of the analysis of the probable directions of faunal flows support the patterns found by both the Parsimony Analysis of Endemicity and the geographical distribution. Four main source areas for rissoids emerge: Mediterranean, Caribbean, Canaries/Madeira archipelagos, and the Cape Verde archipelago. We must stress the high percentage of endemics that occurs in the isolated islands of Saint Helena, Tristan da Cunha, Cape Verde archipelago and also the Azores, thus reinforcing the legislative protective actions that the local governments have implemented in these islands during the recent years.

Conservation genetics of the endemic Azorean slug Plutonia atlantica (Mollusca, Pulmonata)

Abstract Fifty-six specimens of the rare and endangered, endemic Azorean carnivorous slug, Plutonia atlantica , were surveyed for variation at 13 putative allozyme loci in eight populations from three islands. Genotypic proportions showed no significant deviations from Hardy–Weinberg equilibrium expectations and average observed and expected heterozygosity values were H o =0.109 and H e =0.118. At nearly all loci there were significant allele frequency differences between populations. Hierarchical F -statistics, genetic distances (identities), allelic distributions, and an independent analysis of isoelectric focused esterase profiles, suggested that genetic variation in P. atlantica is geographically structured, i.e. within the island of Sao Miguel (Tronqueira region vs the remainder of the island), between islands and between island groups (Faial/Pico vs. Sao Miguel). Genetic distances between populations from any of these geographic units were typical for interspecific comparisons. Therefore, it was concluded that conservation efforts in P. atlantica should recognize at least four management units (Faial, Pico, Tronqueira in Sao Miguel and the remainder of Sao Miguel), in order to effectively preserve the currently known genetic diversity in this species. How far these management units represent evolutionarily significant units or even different taxa remains to be investigated.

Assessing the efficiency of protected areas to represent biodiversity: a small island case study

Protected areas (PAs) have been selected using either subjective or objective criteria applied to an extremely limited subset of biodiversity. Improved availability of species distribution data, better statistical tools to predict species distributions and algorithms to optimize spatial conservation planning allow many impediments to be overcome, particularly on small islands. This study analyses whether 219 species are adequately protected by PAs on Pico Island (the Azores, Portugal), and if they are as efficient as possible, maximizing species protection while minimizing costs. We performed distribution modelling of species’ potential distributions, proposed individual conservation targets (considering the context of each species in the archipelago and their current conservation status) to determine the efficiency of current PAs in meeting such targets and identify alternative or complementary areas relevant for conservation. Results showed that current PAs do not cover all taxa, leaving out important areas for conservation. We demonstrate that by using optimization algorithms it is possible to include most species groups in spatial conservation planning in the Azores with the current resources. With increasing availability of data and methods, this approach could be readily extended to other islands and regions with high endemism levels.

A new species of Pythia Röding, 1798 (Pulmonata, Ellobiidae), from New Ireland, Papua New Guinea

Pythia colmani, a new species from the rainforest of New Ireland, Papua New Guinea, is described. The anatomy of the reproductive and nervous systems is studied in detail. The reproductive system agrees with the patterns observed for other species of the genus. The central nervous system, however, presents a slight divergence from patterns previously recognized in the Pythiinae; the right parieto-visceral connective, in the new species, is less than half the length of its left counterpart, whereas in other pythiine species it is slightly longer than half the length of the left parieto-visceral connective. Taking into consideration that the length of the right parieto-visceral connective is nevertheless appreciable, the significance of this deviation from previously recognized diagnostic characters of subfamilial division of the Ellobiidae is minimized . The new species is compared to Pythia scarabaeus (Linnaeus, 1758), P. pyramidata (Reeve, 1842), P. argenvillei Pfeiffer, 1853 and P. savaiensis Mousson, 1...

Mitochondrial DNA hyperdiversity and its potential causes in the marine periwinkle Melarhaphe neritoides (Mollusca: Gastropoda)

We report the presence of mitochondrial DNA (mtDNA) hyperdiversity in the marine periwinkle Melarhaphe neritoides (Linnaeus, 1758), the first such case among marine gastropods. Our dataset consisted of concatenated 16S-COI-Cytb gene fragments. We used Bayesian analyses to investigate three putative causes underlying genetic variation, and estimated the mtDNA mutation rate, possible signatures of selection and the effective population size of the species in the Azores archipelago. The mtDNA hyperdiversity in M. neritoides is characterized by extremely high haplotype diversity (Hd = 0.999 ± 0.001), high nucleotide diversity (π = 0.013 ± 0.001), and neutral nucleotide diversity above the threshold of 5% (πsyn = 0.0677). Haplotype richness is very high even at spatial scales as small as 100m2. Yet, mtDNA hyperdiversity does not affect the ability of DNA barcoding to identify M. neritoides. The mtDNA hyperdiversity in M. neritoides is best explained by the remarkably high mutation rate at the COI locus (μ = 5.82 × 10−5 per site per year or μ = 1.99 × 10−4 mutations per nucleotide site per generation), whereas the effective population size of this planktonic-dispersing species is surprisingly small (Ne = 5, 256; CI = 1,312–3,7495) probably due to the putative influence of selection. Comparison with COI nucleotide diversity values in other organisms suggests that mtDNA hyperdiversity may be more frequently linked to high μ values and that mtDNA hyperdiversity may be more common across other phyla than currently appreciated.

Assessing the efficiency of protected areas to represent biodiversity: a small island case study – CORRIGENDUM

Marta Vergílio would like to thank the Azorean Regional Fund for Science and Technology and the Pro-Emprego for funding the PhD Project M3.1.2/F/007/2011. The authors would like to thank the Portuguese Foundation for Science and Technology (FCT), for funding the project PTDC/AAC-AMB/098786/2008. This work was also funded by FEDER funds through the Operational Programme for Competitiveness Factors COMPETE and by National Funds through FCT Foundation for Science and Technology under the project No. FCOMP-01-0124FEDER-037300 (Ref. FCT PEstC / BIA / UI0609 / 2013), UID/BIA/50027/2013, and POCI-01-0145-FEDER00682. Data on species distributions was gathered based on the European Union projects INTERREGIII B “ATLÂNTICO” (2004–2006) and BIONATURA (2006– 2008), and, more recently, ATLANTISMAR “Mapping coastal and marine biodiversity of the Azores” (Ref: M2.1.2/I/027/2011). The climatic modeling work of EBA was developed in the framework of the project “EstraMAR” (MAC/3/C177), which was supported by the European Union through the MAC Transnational Program of Cooperation – Madeira-Azores-Canaries.