Chromosomes of three freshwater stingrays (Rajiformes Potamotrygonidae) from the Rio Negro basin, Amazon, Brazil

Potamotrygonidae is the representative family of South American freshwater elasmobranchs. It is a monophyletic group containing 20 species grouped into three genera. Three species belonging to two genera of this family were collected from the middle Negro River, Amazonas, Brazil, and studied cytogenetically: Paratrygon aiereba, Potamotrygon motoro and Potamotrygon orbignyi. Paratrygon aiereba presented 2n = 90 chromosomes and 4M+2SM+10ST+74A. Both species of Potamotrygon presented 2n = 66 chromosomes and differed in their chromosomal formulas: P. motoro had 18M+12SM+10ST+26A and P. orbignyi had 22M+10SM+8ST+26A. No sex heteromorphism was detected. The Fundamental Number (FN) was 106 for the three species. A system of multiple NORs was found in the three species, but with interspecific differences in terms of location and position of the active Ag-NORs sites. Paratrygon aiereba presented only four sites on the short arms of two chromosomal pairs, both in terminal regions. Potamotrygon motoro presented seven sites, on the long and short arms, all in terminal regions of non-homologous chromosomes; P. orbignyi presented eight sites on the long arms, all in terminal regions, of non-homologous chromosomes. The constitutive heterochromatin was in pericentromeric regions of all chromosomes, and no significant interspecific difference was found in relation to this marker.     

Taxonomic revision of Rhinebothroides (Eucestoda: Tetraphyllidea: Phyllobothriidae), parasites of neotropical freshwater stingrays (Rajiformes: Myliobatoidei: Potamotrygonidae)

Extensive new collections of specimens of Rhinebothroides spp. from a wide range of Neotropical freshwater potamotrygonid stingrays in numerous localities permitted critical evaluation of the validity of the 7 nominal species in the genus. Some qualitative characters previously used in the taxonomy of this group are highly variable within and among populations, and most morphometric and meristic characters thought to have taxonomic value for this group overlapped among several previously defined nominal species. Characters that were consistent throughout populations and that clearly define morphotypes supported recognition of 4 species: R. freitasi (syn. R. circularisi and R. venezuelensis), R. glandularis (syn. R. mclennanae), R. moralaurai, and R. scorzai. We provide a taxonomic review of the genus, 8 additional host records, 7 new localities, and an identification key.     

Origins, diversification, and historical structure of the helminth fauna inhabiting neotropical freshwater stingrays (Potamotrygonidae).

Members of the freshwater stingray family Potamotrygonidae occur throughout the major river systems of eastern South America that empty into the Atlantic Ocean. Ichthyologists have tended to assume that the ancestor of the potamotrygonids was an Atlantic marine or euryhaline stingray that dispersed into freshwater, presumably during the last marine ingression 3-5 million years ago. The helminth parasites that inhabit potamotrygonids suggest an alternative perspective on their origin. Phylogenetic and biogeographic analysis of the helminths inhabiting potamotrygonids suggest that the hosts are derived from an ancestral Pacific urolophid stingray that was trapped in freshwater by the uplifting of the Andes beginning perhaps as early as the early Cretaceous period and ending by the mid-Miocene epoch, changing the course of the Amazon River, which previously had flowed into the Pacific Ocean.     

Himantura lobistoma, a new whipray (Rajiformes: Dasyatidae) from Borneo, with comments on the status of Dasyatis microphthalmus

A new long-snouted whipray, Himantura lobistoma, is described on the basis of seven specimens collected from the southeastern sector of the South China Sea, off western Borneo. It belongs to a subgroup of Himantura species provisionally defined herein as the “uarnacoides” complex that includes five nominal species: H. chaophraya, H. granulata, H. hortlei, H. pastinacoides, and H. uarnacoides. These whiprays all have a similar lateral disc shape (i.e., pectoral-fin apices broadly rounded rather than narrowly rounded or angular), a plain colouration (i.e., usually uniform light to dark brown dorsally, and pale or whitish ventrally with or without a dark margin), a long and slender whiplike tail, and platelike denticles in a broad dorsal band in adults. Himantura lobistoma can be distinguished from the superficially similar H. hortlei by its highly protrusible mouth and pelvic girdle with prominent lateral prepelvic processes. It differs from other members of the “uarnacoides” complex by a combination of internal and external morphological characteristics (e.g., having an extremely elongate snout and very small eyes). Another long-snouted stingray from the region, Dasyatis microphthalmus Chen, inadequately described and known only from the lost holotype, is considered a nomen dubium. It had been assigned to Himantura based on the absence of an obvious ventral cutaneous tailfold but may be conspecific with D. acutirostra.     

Systematics, cladistics and biogeography of the American genus Farrodes (Ephemeroptera: Leptophlebiidae: Atalophlebiinae)

Cladistic and biogeographic analyses of the genus Farrodes are presented. Two species groups are delineated within Farrodes: F. caribbianus and F. bimaculatus. Three species formerly placed in other genera– Thraulus caribbianus Traver, Thraulus roundsi Traver and Homothraulus maculatus (Needham & Murphy)–are transferred to Farrodes. The species of the F. caribbianus species group are revised. Three new species are described: F. savagei from Venezuela, F. maya and F. mexicanus from Mexico. Keys to separate the two species groups of Farrodes and the species of the F caribbianus species group are provided. Successive cladistic analyses were carried out on both adult and nymphal characters using Hennig86 and CLADOS. The matrix was composed of all available data (nymphal characters were missing for some species), from nymphal and adult stages separately and on taxa represented by both adult and nymphal characters. Species of the genera Simothraulopsis and Homothraulus (components of the Farrodes lineage) were included in the analyses, and Ecuaphkbia was used as the outgroup. Results of the four analyses are compatible. The historical biogeography of Farrodes , with a distribution from nordiern Argentina to southern Texas, is analysed using the program COMPONENT. Areas of endemism are established, and some of their relationships compared with those of other groups available in the literature.     

A re‐examination of the molecular phylogeny and biogeography of the genus Schizothorax (Teleostei: Cyprinidae) through enhanced sampling,with emphasis on the species in the Yunnan–Guizhou Plateau,China

The mitochondrial DNA cytochrome b sequences of 36 Schizothorax species from 51 localities in the Yunnan–Guizhou Plateau (YGP) and its adjacent areas were analysed. Maximum parsimony, Maximum likelihood and Bayesian phylogenetic analyses were performed to examine the relationships of Schizothorax species. A hypothesis of the phylogenetic relationships of the species is given. A relaxed molecular clock based on Bayesian evolutionary analysis was used to tentatively calculate the divergence times of Schizothorax. Samples from the YGP were tentatively grouped into three geographically distributed clades: the Tsangpo‐Irrawaddy, the Mekong‐Salween and the Trans‐Jinsha River (including Jinsha, Red, Nanpan and Beipan Rivers). Calibration of the molecular clock revealed that two geological periods, the late Miocene about 10 million years before present (Myr BP) and the Pliocene (4.0 Myr BP), were important times in the vicariant speciation of Schizothorax. The phylogenetic history of the species is congruent with events caused by the uplift of the Tibetan Plateau and the YGP. The divergence of Schizothorax species in YGP began in the Pliocene. Our phylogenetic trees did not support the hypothesis that the paleo Jinsha River was drained through the Yangtze River‐Jianchuan Lake‐Erhai Lake to the Red River. Schizothorax in the Beipan River were derived from the Jinsha River.     

Molecular systematics,taxonomy and biogeography of the genus Cavia (Rodentia: Caviidae)

Phylogenetic analyses were conducted on cytochrome b sequence data of the most geographically and taxonomically broad sampling of Cavia taxa to date. Primary objectives included providing the first extensive molecular phylogenetic framework for the genus, testing the taxonomic and systematic hypotheses of previous authors and providing insight into the evolutionary and biogeographic history of the genus. Support was found for the morphologically defined species C. aperea, C. tschudii, C. magna and C. fulgida and the taxonomic placement of taxa previously subject to conflicting taxonomic opinions (e.g. C. nana, C. anolaimae and C. guianae) was further resolved. Additionally, we elevate the Ecuadorian C. a. patzelti to species status, restrict the distributional limits and suggest taxonomic affiliations of some C. tschudii subspecies, and provide strong evidence for the geographic origin of guinea pig domestication. Finally, we provide an estimated evolutionary timeline for the genus Cavia, which appears to extend well into the late Miocene.     

Molecular phylogeny and the historical biogeography of the warblers of the genus Sylvia (Aves)

A molecular phylogeny based on DNA/DNA hybridization revealed that the Sylvia-Parisoma complex is monophyletic and includes three main groups of species, the “mid-European” warblers, the genus Parisoma, and the “eu-Mediterranean” Sylvia species sensu stricto. The latter can be assigned to three main clusters, a “West-Mediterranean” group, a “Central-Mediterranean group”, and an “East-Mediterranean” group. The radiation of the whole complex is much more ancient than formerly believed. It started ca 12–13 Ma ago and the ancestors of the main extant groups differentiated during the Pliocene. Only speciation events within the “eu-Mediterranean” lineages occurred during the Pleistocene. The paleoclimatical and paleoecological history of the Mediterranean region is too complicated to provide any evidence for direct relationships between past events and evolutionary steps of these taxa which did not leave any reliable fossil record. However, some major speciation events may be related to well documented climatical crises as well as paleobotanical data. The largely man-induced extension of matorrals over several millenia presumably extended the range of several species that were formerly much more restricted, which complicates reconstruction of the spatio-temporal course of speciation.     

Molecular phylogeny and historical biogeography of the Anatolian lizard Apathya (Squamata,Lacertidae)

Apathya is a lacertid genus occurring mainly in south-east Turkey and its adjacent regions (part of Iran and Iraq). So far two morphological species have been attributed to the genus; A. cappadocica (with five subspecies, A. c. cappadocica, A. c. muhtari, A. c. schmidtlerorum, A. c. urmiana and A. c. wolteri) and A. yassujica. The first species occupies most of the genus’ distribution range, while A. yassujica is endemic of the Zagros Mountains. Here, we explored Apathya’s taxonomy and investigated the evolutionary history of the species by employing phylogenetic and phylogeographic approaches and using both mitochondrial (mtDNA) and nuclear markers. The phylogenetic relationships and the genetic distances retrieved, revealed that Apathya is a highly variable genus, which parallels its high morphological variation. Such levels of morphological and genetic differentiation often exceed those between species of other Lacertini genera that are already treated as full species, suggesting the necessity for a taxonomic revision of Apathya. The phylogeographical scenario emerging from the genetic data suggests that the present distribution of the genus was determined by a combination of dispersal and vicariance events between Anatolia and Southwest Asia dating back to the Miocene and continuing up to the Pleistocene. Key geological events for the understanding of the phylogeography of the genus are the movement of the Arabian plate that led to the configuration of Middle East (orogenesis of the mountain ranges of Turkey and Iran) and the formation of Anatolian Diagonal.     

Phylogeny and biogeography of Valerianaceae (Dipsacales) with special reference to the South American valerians

Species of Valerianaceae are a common component of the alpine flora throughout the Northern Hemisphere as well as the Andes of South America. Sequence data from three chloroplast markers (psbA-trnH intron, trnK-matK intron, and the trnL-F region) along with the internal transcribed spacer region (ITS) of nuclear ribosomal DNA were used to infer relationships within Valerianaceae. Both genomes, as well as a combined data set, provide support for the major clades within the group and do not support a monophyletic Valeriana. In addition, these data indicate that Plectritis is nested within South American Valeriana, as opposed to being sister to Centhranthus as previously hypothesized. Valerianaceae appear to have originated in Asia, probably in the Himalayas, and subsequently to have dispersed several times to Europe and to the New World. Our results imply that Valerianaceae colonized South America on multiple occasions from the north. In one of these cases there appears to have been a substantial and rapid radiation, primarily in the high elevation paramo habitat. A variety of methods were used to estimate divergence times to determine when Valerianaceae might have colonized South America. Regardless of the method and fossil constraints applied, our estimates suggest that Valerianaceae colonized South America prior to the formation of the Isthmus of Panama.     

Revision of Potamotrygonocotyle Mayes, Brooks & Thorson, 1981 (Platyhelminthes: Monogenoidea: Monocotylidae), with descriptions of four new species from the gills of the freshwater stingrays Potamotrygon spp. (Rajiformes: Potamotrygonidae) from the La Plata river basin

The only known monocotylid genus to parasitise Neotropical freshwater stingrays (Potamotrygonidae) is Potamotrygonocotyle Mayes, Brooks & Thorson, 1981, a monotypic genus erected to accommodate P. tsalickisi Mayes, Brooks & Thorson, 1981. For more than 20 years, no other species has been recognised in this genus, but new efforts to survey the diversity of parasites inhabiting potamotrygonids have revealed the existence of new species and the need to redefine the genus. Here, the generic diagnosis of Potamotrygonocotyle is amended, P. tsalickisi is redescribed and four new species are recognised and described based on samples collected from the gills of freshwater potamotrygonids from the La Plata river basin: Potamotrygonocotyle chisholmae n. sp. and P. dromedarius n. sp. from Potamotrygon motoro; Potamotrygonocotyle eurypotamoxenus n. sp. from Potamotrygon cf. motoro (type-host), P. castexi, P. falkneri and P. histrix; and Potamotrygonocotyle uruguayensis n. sp. from Potamotrygon brachyura. Potamotrygonocotyle is characterised by species possessing: (1) slightly sinuous sclerotised ridges on all septa; (2) two pairs of the dorsal haptoral accessory structures associated with the four posterior peripheral loculi and with anterior dorsal haptoral accessory structure bilobate or semicircular; and (3) male copulatory organ without an accessory piece.     

Systematics and phylogeny of the Aglajidae (Opisthobranchia: Mollusca)

The generic limits within the Aglajidae are reviewed and modified, based on the study of 21 species. Navanax Pilsbry, 1895 is reinstated as a genus distinct from Aglaja. The generic placement of several species is altered from previous works. An evaluation of morphological characters suggests that each of six genera has distinctive external morphology. The systematic value of twelve characters is critically reviewed. The evolutionary relationships of the Aglajidae and its genera to other philinacean familes are discussed.     

Systematics and biogeography of the Neotropical genus Mabuya,with special emphasis on the Amazonian skink Mabuya nigropunctata (Reptilia,Scincidae)

Phylogenetic analyses using up to 1532 base pairs (bp) of mitochondrial DNA from 106 specimens of Neotropical Mabuya, including 18 of the 19 recognized South American and Mesoamerican species, indicate that most species of the genus are monophyletic, including M. nigropunctata that had previously been reported to be paraphyletic. The present results shows that this species includes three highly divergent and largely allopatric lineages restricted to occidental, meridional, and oriental Amazonia. Our dataset demonstrates that previous claims regarding the paraphyletic status of M. nigropunctata and the phylogenetic relationships within this species complex based on the analysis of three mitochondrial and four nuclear genes (approx. 5000 bp) were erroneous and resulted from two contaminated cytochrome b sequences.The phylogenetic results indicate that diversification in the Neotropical genus Mabuya started approximately in the Middle Miocene (15.5–13.4 Ma). The divergence dates estimated for the Mabuya nigropunctata species complex suggest that the major cladogenetic events that produced the three main groups (occidental (oriental + meridional)) occurred during the Late Miocene. These estimations show that diversification within the M. nigropunctata species complex was not triggered by the climatic changes that occurred during the Pleistocene, as has been suggested by several authors. Rather, our data support the hypothesis that the late tertiary (essentially Miocene epoch) was a period that played a very important role in the generation of biological diversity in the Amazonian forests.Speciation between Mabuya carvalhoi, endemic to the coastal mountain range of Venezuela, and M. croizati, restricted to the Guiana Shield, occurred during the Middle Miocene and may have been as the result of a vicariant event produced by the formation of the present day Orinoco river drainage basin and the consequent appearance of the Llanos del Orinoco, which acted as a barrier to dispersal between these two species. The split between M. bistriata and M. altamazonica and between the occidental and (meridional + oriental) clades of M. nigropunctata fits very well with the biogeographic split between the eastern and western Amazon basins reported for several other taxa.     

Cestode parasites in Potamotrygon motoro (Natterer) (Chondrichthyes: Potamotrygonidae) from southwestern Brazil, including Rhinebothroides mclennanae n. sp. (Tetraphyllidea: Phyllobothriidae), and a revised host-parasite checklist for helminths inhabiting neotropical freshwater stingrays.

Specimens of 5 species of cestodes were collected in 6 specimens of the freshwater stingray species Potamotrygon motoro (Natterer), collected in the vicinity of Corumba, Mato Grosso do Sul, Brazil. Acanthobothrium regoi, Potamotrygonocestus orinocoensis, Rhinebothroides venezuelensis, and Rhinebothrium paratrygoni are reported from P. motoro and from southwestern Brazil for the first time. Rhinebothroides mclennanae n. sp. appears to be the sister species of Rhinebothroides glandularis, the only other member of the genus exhibiting darkly staining glandular cells lying free in the parenchyma surrounding the terminal genitalia. The new species resembles Rhinebothroides glandularis, Rhinebothroides freitasi, and Rhinebothroides scorzai by having poral ovarian arms that extend anteriorly beyond the posterior margin of the cirrus sac, coiled vaginae, and vitelline follicles not interrupted on the poral side in the vicinity of the genital pore. It differs from all 6 previously described members of the genus by possessing an average of 31 testes per proglottid, compared with an average of 45 for R. glandularis, 55 for R. freitasi and R. venezuelensis, 77 for Rhinebothroides circularisi and Rhinebothroides moralarai, and 80 for R. scorzai. An updated phylogenetic tree for Rhinebothroides is presented.     

Systematics and biogeography of the jellyfish Aurelia labiata (Cnidaria: Scyphozoa)

The hypothesis that the common eastern North Pacific Aurelia is A. aurita is falsified with morphological analysis. The name Aurelia labiata is resurrected, and the species is redescribed, to refer to medusae differing from A. aurita by a suite of characters related to a broad and elongated manubrium. Specifically, the oral arms are short, separated by and arising from the base of the fleshy manubrium, and the planulae are brooded upon the manubrium itself, rather than on the oral arms. Aurelia aurita possesses no corresponding enlarged structure. Furthermore, the number of radial canals is typically much greater in A. labiata, and thus the canals often appear more anastomosed than in A. aurita. Finally, most A. labiata medusae possess a 16-scalloped bell margin, whereas the margin is 8-scalloped in most A. aurita. Separation of the two forms has previously been noted on the basis of allozyme and isozyme analyses and on the histology of the neuromuscular system. Partial 18S rDNA sequencing corroborates these findings. Three distinct morphotypes of A. labiata, corresponding to separate marine bioprovinces, have been identified among 17 populations from San Diego, California, to Prince William Sound, Alaska. The long-undisputed species A. limbata may be simply a color morph of A. labiata, or a species within a yet-unelaborated A. labiata species complex. The first known introduction of Aurelia cf. aurita into southern California waters is documented. Although traditional jellyfish taxonomy tends to recognize many species as cosmopolitan or nearly so, these results indicate that coastal species, such as A. labiata, may experience rapid divergence among isolated populations, and that the taxonomy of such species should therefore be scrutinized with special care.     

Molecular phylogeny and biogeography of an ancient Holarctic lineage of mygalomorph spiders (Araneae: Antrodiaetidae: Antrodiaetus)

The mygalomorph spider genera Antrodiaetus and Atypoides (Antrodiaetidae) belong to an ancient lineage that has persisted since at least the Cretaceous. These spiders display a classic disjunct Holarctic distribution with species in the eastern Palaearctic plus the western and eastern Nearctic. Prior phylogenetic analyses of this group have been proposed on the basis of morphology, but lack strong support and independent corroboration. Here we present the first phylogenetic analysis of species-level relationships based on molecular data obtained from the mitochondrial (cytochrome c oxidase subunit I) and nuclear (18S and 28S rRNA) genomes. Analyses corroborate earlier findings that Atypoides forms a paraphyletic grade with respect to Antrodiaetus, and consequently, that genus is formally synonymized under Antrodiaetus. In addition, our results support the relatively early divergence of Antrodiaetus roretzi. Antrodiaetus pacificus is "paraphyletic" with respect to the A. lincolnianus group and is likely an assemblage of numerous species. The final topology based on a combined molecular dataset, in conjunction with two different molecular dating techniques (penalized likelihood plus a Bayesian approach) and ancestral distribution reconstructions, was used to infer the historical biogeography of these spiders. Trans-Beringian and trans-Atlantic routes appear to account for the present-day distribution of Antrodiaetus in Japan and North America. Future studies on Antrodiaetus phylogeny will be used to address questions regarding morphological stasis and the evolution of quantitative morphological characters.     

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.