Computational protocol: Biogeography and phylogenetic position of Enchodeloidessignyensis (Loof, 1975), gen. n., comb. n. from Maritime Antarctic (Nematoda, Nordiidae)

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[…] Samples were collected from Livingston Island by Dr. N. Chipev (IBER), Dr. R. Mecheva (IBER), D. Apostolova (Sofia University) and from King George Island by Dr. R. Zidarova (Sofia University) during the regular Bulgarian Antarctic Expeditions (2006-2016). Nematodes were extracted from soils and plant materials by a Baerman funnel method () for at least 48 hours, killed by gentle heat, and fixed in 4% formalin. For light-microscopy, specimens were processed in anhydrous glycerine () and mounted on permanent slides. Drawings were prepared using an Olympus BX 51 compound microscope, equipped with a drawing tube. Photographs were taken using an Axio Imager.M2-Carl Zeiss compound microscope equipped with a digital camera (ProgRes C7) and specialised software (CapturePro Software 2.8). Measurements were made using an Olympus BX 41 light microscope with a drawing tube and digitising tablet (CalComp Drawing Board III, GTCO CalCom Peripherals, Scottsdale, AZ, USA) and Digitrak 1.0f computer program (Philip Smith, John Hutton Institute, Dundee, UK).Specimens used for SEM observations were rinsed in 0.1 M cacodylate buffer (twice for 10 min), post-fixed in 1% OsO4 for 2 h, washed twice for 10 min in 0.1 M cacodylate buffer and dehydrated in an ethanol series (), immersed in hexamethyldisilazane for 30 min and air dried. They were sputter coated with gold in a JEOL JFS 1200 and examined using a JEOL JSM 5510 microscope at 10 kV.The locations of pharyngeal gland nuclei are given following and . [...] The 18S and D2-D3 28S rDNA sequences were compared with those of other nematode species available at the GenBank sequence database using BLASTN similarity search tool. The sequences revealing the highest similarity were used for sequence and phylogenetic analyses (; ; , ; ; , etc.). Bayesian Inference (BI) algorithm implemented in MrBayes 3.2.5 was used for reconstruction of phylogenetic relationships (; ). For further details on phylogeny analyses and tree visualisation, see . Based on previous studies (; ) Aporcelaimellus spp. were selected as an outgroup for both phylogenies. The estimates of evolutionary divergences between sequences/species within and between groups (numbers of base differences and p-distances) were performed with MEGA7 (). The analyses involved nine nucleotide sequences with 790 and 1666 positions in total for D2-D3 and 18S rDNA, respectively. […]

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