Rhabdias glaurungi sp. nov. (Nematoda: Rhabdiasidae), parasite of Scinax gr. ruber (Laurenti, 1768) (Anura: Hylidae), from the Brazilian Amazon.

The genus Rhabdias Stiles & Hassal, 1905 includes about 83 species of nematodes parasitic in amphibians and reptiles worldwide. Herein, we describe Rhabdias glaurungi sp. nov. from the hylid frog Scinax gr. ruber (Laurenti, 1768) in the Gunma Ecological Park, Santa Bárbara municipality, state of Pará, Brazil. This species has six small lips, an inflated cuticle along the entire body and a cup-shaped buccal capsule with smooth internal surface of its anterior part and irregularly folded internal surface of its posterior part in apical view. From the 17 valid species recognized in the Neotropical realm, the new species can be distinguished by the number of lips, the morphology and size of its buccal capsule, as well as the extent and shape of its cuticular inflation; in addition, there are molecular differences. Sequences of the mitochondrial Cytochrome c Oxidase subunit I gene strongly support the status of this form as a separate species. Molecular phylogenetic analysis shows R. glaurungi sp. nov. nested within the R. pseudosphaerocephala Kuzmin, Tkach & Brooks, 2007 species complex. Rhabdias glaurungi sp. nov. is the second species of the genus described from hosts of the family Hylidae in the Neotropical realm. We conclude that the diversity of Rhabdias within the Neotropics is likely largely underestimated.

Description, molecular characterization and life cycle of Serpentirhabdias mussuranae n. sp. (Nematoda: Rhabdiasidae) from Clelia clelia (Reptilia: Colubroidea) in Brazil.

Serpentirhabdias mussuranae n. sp. is described from the lungs of the mussurana, Clelia clelia (Daudin, 1803), from vicinities of Lábrea, Amazonas State, Brazil. The species is characterized by the triangular oral opening, the presence of teeth (onchia) in the oesophastome, the excretory glands longer than the oesophagus and the tail abruptly narrowing in its anterior half and gradually tapering in posterior half. Among the Neotropical representatives of the genus, three species are known to possess the onchia in the oesophastome: S. atroxi, S. moi and S. viperidicus. Serpentirhabdias mussuranae n. sp. differs from S. atroxi and S. viperidicus by its triangular shape of the oral opening and the oesophastome in apical view, vs. round in the latter two congeners. Additionally, S. viperidicus has a larger oesophastome, 13-22 micrometers wide and 13-23 micrometers deep. The new species has relatively longer excretory glands than S. moi. The new species is morphologically and genetically close to S. atroxi, S. moi and S. viperidicus, all parasitic in Brazilian snakes, based on the presence of onchia and the comparison of nucleotide sequences of nuclear ribosomal DNA and mitochondrial cox1 gene (differences varied between 3.8% and 7.1%). Data on the life cycle of S. mussuranae n. sp. is provided, and the life cycle is typical of the genus Serpentirhabdias, with the combination of direct development and heterogony. Free-living larval stages and the adults of amphimictic free-living generation are described. The results of molecular phylogenetic analysis based on nuclear ribosomal internal transcribed spacer (ITS) + partial 28S region and partial mitochondrial cox1 gene are provided.

Ecological, morphological, and molecular studies of Acanthocheilonema odendhali (Nematoda: Filarioidea) in northern fur seals (Callorhinus ursinus) on St. Paul Island, Alaska.

Studies of northern fur seal (Callorhinus ursinus Linnaeus, 1758) infection by the filariid nematode Acanthocheilonema odendhali were carried out in 2011-2012 on St. Paul Island, Pribilof Archipelago, Alaska. Skins of 502 humanely harvested northern fur seals from haul-out areas of five rookeries, Polovina (n = 122), Morjovi (n = 54), Zapadni (n = 72), Lukanin (n = 109), and Gorbatch (n = 145), were examined. A. odendhali was found in 18% of northern fur seals. The prevalence of infection ranged from 12.5% up to 22.9% on different haul-out areas on the island. The mean intensity of infection was 1.3 (range 1-7). Detailed morphological examination of collected specimens was performed using light microscopy. Several characters were added to the morphological description of the species, among them lateral thickening of the body cuticle, especially prominent in males, variations in number and position of genital papillae in males, transverse striation of the cuticle, and terminal dilation on tail end in microfilariae. The adult specimens studied had a shorter esophagus than type specimens from the California sea lion described by Perry (1967). Comparison of partial sequences of the mitochondrial cox1 gene from specimens collected from five sampling sites on St. Paul Island and a specimen from the type host and territory in California showed no significant differences and strongly supported conspecificity of the material from Alaska with A. odendhali.

Pattern of extinction of the woolly mammoth in Beringia.

Extinction of the woolly mammoth in Beringia has long been subject to research and speculation. Here we use a new geo-referenced database of radiocarbon-dated evidence to show that mammoths were abundant in the open-habitat of Marine Isotope Stage 3 (∼45-30 ka). During the Last Glacial Maximum (∼25-20 ka), northern populations declined while those in interior Siberia increased. Northern mammoths increased after the glacial maximum, but declined at and after the Younger Dryas (∼12.9-11.5 ka). Remaining continental mammoths, now concentrated in the north, disappeared in the early Holocene with development of extensive peatlands, wet tundra, birch shrubland and coniferous forest. Long sympatry in Siberia suggests that humans may be best seen as a synergistic cofactor in that extirpation. The extinction of island populations occurred at ∼4 ka. Mammoth extinction was not due to a single cause, but followed a long trajectory in concert with changes in climate, habitat and human presence.

Patterns of nucleotide misincorporations during enzymatic amplification and direct large-scale sequencing of ancient DNA.

Whereas evolutionary inferences derived from present-day DNA sequences are by necessity indirect, ancient DNA sequences provide a direct view of past genetic variants. However, base lesions that accumulate in DNA over time may cause nucleotide misincorporations when ancient DNA sequences are replicated. By repeated amplifications of mitochondrial DNA sequences from a large number of ancient wolf remains, we show that C/G-to-T/A transitions are the predominant type of such misincorporations. Using a massively parallel sequencing method that allows large numbers of single DNA strands to be sequenced, we show that modifications of C, as well as to a lesser extent of G, residues cause such misincorporations. Experiments where oligonucleotides containing modified bases are used as templates in amplification reactions suggest that both of these types of misincorporations can be caused by deamination of the template bases. New DNA sequencing methods in conjunction with knowledge of misincorporation processes have now, in principle, opened the way for the determination of complete genomes from organisms that became extinct during and after the last glaciation.

Field study on the survival, migration and overwintering of infective larvae of horse strongyles on pasture in central Ukraine.

Experimental studies on the survival of infective stage larvae of horse strongyles and their ability to overwinter on pasture were carried out in central Ukraine (Poltavska oblast). Faecal pats (1.5 kg each) of naturally infected horses were placed on pasture, and samples of faeces and surrounding vegetation (10 g each) were collected each month, excluding the winter months, from November 2002 until April 2004. The number of infective third stage larvae was calculated in each sample and compared with that from the control faecal samples cultivated in the laboratory. In the control samples, the ratio of infective third stage larvae to the initial number of eggs was from 54.7% in June up to 84.2% in November. This ratio depended on the presence of nematophagous fungi growing in the faeces. On pasture, the development of larvae to the infective third stage took approximately 4 weeks in the warm season, from April until September. In October, a percentage of the eggs (25% to EPG value) did not hatch. No larval development was observed in faeces in November. A minute quantity of larvae, about 0.03% of their initial number, was observed to survive on pasture for the 12 months. Migration of infective larvae from the faeces to vegetation was not intensive, between 71% and 89% of larvae remained in the faeces 4 weeks after deposition of the faecal pats, the percentage related to soil humidity in each month. The proportion of larvae successfully surviving during winter appeared to be maximal in faecal pats deposited on pasture in September of the previous year (up to 42.0% of the initial number of larvae). Some larvae were observed surviving winter in soil beneath the faecal pats. The results of the study demonstrated that horse pastures in the central part of Ukraine are never free from the infective third stage larvae of strongyles.

Helminths of the sand lizard, Lacerta agilis (Reptilia, Lacertidae), in the Palaearctic: faunal diversity and spatial patterns of variation in the composition and structure of component communities.

We studied variation in the structure of component communities of helminths in sand lizards, Lacerta agilis, from 30 localities in the Ukraine and Bulgaria. Thirty-five separate samples of lizards, with a total of 661 completely censused infracommunities, yielded 30 helminth species (4 cestodes, 10 trematodes, 3 acanthocephalans and 13 nematodes). In its range within the Ukraine, L. agilis serves as the final host for 13 species of which only 3 (S. lacertae, S. hoffmanni and P. molini) can be considered as lizard specialists. A characteristic feature of these helminth component communities was the large proportion of heteroxeneous helminth species for which L. agilis serves as paratenic host. Sand lizards in the meadow steppeland zone were primarily parasitized by larval helminths that represented a major proportion of the total number of all worms recovered while those sampled in the grassland/forest transition zone were characterized by substantially higher proportions of adult helminths using lizards as final hosts. However, L. agilis was parasitized by a much higher proportion of lizard specialists in the 'typical' habitats of the meadow steppeland zone as opposed to those located in 'marginal' habitats in the grassland/forest transition zone, where helminths were shared to a greater extent with amphibian hosts.