An easy and low-cost method to elaborate a container to dehydrate nematodes and other meiofauna in order to process them for scanning electron microscopy (SEM) is presented. Illustrations of its elaboration, step by step, are included. In addition, a brief methodology to process meiofauna, especially nematodes and kinorhynchs, and illustrations are provided. With this methodology it is possible to easily introduce the specimens, to lock them in a closed chamber allowing the infiltration of fluids and gases (ethanol, acetone, carbon dioxide) but avoiding losing the specimens. After using this meiofauna basket for SEM the results are efficient. Examples of nematode and kinorhynch SEM pictures obtained using this methodology are also included.
Species of the nematode genus Heterorhabditis are important biological control agents against agricultural pests. The taxonomy of this group is still unclear as it currently relies on phylogenetic reconstructions based on a few genetic markers with little resolutive power, specially of closely related species. To fill this knowledge gap, we sequenced several phylogenetically relevant genetic loci and used them to reconstruct phylogenetic trees, to calculate sequence similarity scores, and to determine signatures of species- and population-specific genetic polymorphism. In addition, we revisited the current literature related to the description, synonymisation, and declaration as species inquirendae of Heterorhabditis species to compile taxonomically relevant morphological and morphometric characters, characterized new nematode isolates at the morphological and morphometrical level, and conducted self-crossing and cross-hybridization experiments. The results of this study show that the sequences of the mitochondrial cytochrome C oxidase subunit I (COI) gene provide better phylogenetic resolutive power than the sequences of nuclear rRNA genes and that this gene marker can phylogenetically resolve closely related species and even populations of the same species with high precision. Using this gene marker, we found two new species, Heterorhabditis ruandica n. sp. and Heterorhabditis zacatecana n. sp. A detailed characterization of these species at the morphological and morphometric levels and nematode reproduction assays revealed that the threshold for species delimitation in this genus, using COI sequences, is 97% to 98%. Our study illustrates the importance of rigorous morphological and morphometric characterization and multi-locus sequencing for the description of new species within the genus Heterorhabditis, serves to clarify the phylogenetic relationships of this important group of biological control agents, and can inform future species descriptions to advance our efforts towards developing more tools for sustainable and environmentally friendly agriculture.
A new species of the genus Oscheius, O. saproxylicus sp. n., collected in decaying wood obtained from an orchard in Southern Iberian Peninsula, is reported. A detailed description, including morphometrics, LM and SEM images, and molecular (18S and 28S rDNA genes) information is provided. The female is characterized by a moderately long body, lateral field with three longitudinal ridges, midbody vulva, and conical tail with acute tip. It was distinguished from its closest relative, O. dolichura, by a shorter tail and longer rectum. The male was not found. Morphological and molecular data support its belonging to Dolichura-group. Molecular analyses show that both Insectivorus and Dolichura groups are related to each other, being proposed as subgenera of the genus Oscheius as Oscheius and Dolichorhabditis. Finally, an updated taxonomy of the genus is presented, with generic and subgeneric diagnoses, list of species and a key to their identification.
Two very similar species of the genus Chiloplacus, namely C. magnus and C. insularis, are re-described and compared based on type material as well as Iberian specimens. Relevant new information is obtained for their characterisation and for elucidating their identity with more accuracy. Special mention is deserved by the findings regarding: i) the morphology of the lateral field at vulval level; and ii) the location of the vulva, which is either midventral or subventral, two remarkable traits that were originally overlooked. Chiloplacus magnus is distinguished by having the left lateral field expanded at vulval level and the vulva displaced to a subventral position, whereas C. insularis displays the usual pattern of the genus. The evolutionary relationships of the Iberian C. magnus are explored by means of D2-D3 sequence analysis. Chiloplacus similis is regarded as identical to C. magnus and is proposed as a new junior synonym of the latter.
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