Ainara Achurra

Integrative taxonomy of the freshwater worm Rhyacodrilus falciformis s.l. (Clitellata: Naididae), with the description of a new species

The genetic and morphological variation within Rhyacodrilus falciformis Bretscher, 1901 (Clitellata: Naididae) in Europe was explored using an integrative approach, with three unlinked genetic markers [the mitochondrial cytochrome c oxidase subunit I (COI), the nuclear histone 3 (H3) and internal transcribed spacer region (ITS)] combined with morphology, to investigate whether this taxon constitutes a single or several species. Using Automatic Barcode Gap Discovery on the COI data set, the specimens were divided into seven clusters, used as hypothetical species that were further tested with the other data sources. Single‐gene trees were estimated for all three markers, using coalescence analysis and they were in many parts incongruent with each other. Only one of the clusters was supported by all trees; it was also morphologically differentiated from the other clusters by the shape of its modified penial chaetae. This group consists of two specimens from the Crotot Cave in south‐eastern France, and morphologically they fit a previously described but invalid variety, ‘pigueti’, which is here described as a new species, Rhyacodrilus pigueti Achurra & Martinsson sp. n. The study highlights the fact that a single data source (e.g. COI barcodes) seldom provides a sufficient basis for taxonomic decisions such as species delimitation.

DNA barcoding and species delimitation: the Stylodrilus heringianus case (Annelida : Clitellata : Lumbriculidae)

Abstract. Individuals of the aquatic oligochaete species Stylodrilus heringianus Claparède, 1862 were collected across a part of this species’ distribution range in Sweden, Estonia, Great Britain and Spain to test whether they represent a single metapopulation or several separately evolving lineages. Using sequences of the barcoding gene cytochrome c oxidase subunit I (COI) and two nuclear genes (internal transcribed spacer region and histone 3), three different approaches were conducted: pairwise distance-method, Bayesian inference and network analysis. Both the COI phylogeny and network analyses were concordant in recovering six haplotype clusters, which showed a maximum genetic distance of 7.7% (K2P) among each other. Nevertheless, nuclear genes failed to confirm any lineage separation, and we conclude that the sampled specimens all belong to the same species. A phylogeographic history with allopatric divergence and secondary contact is suggested to explain this intraspecific pattern of mitochondrial divergence and nuclear non-divergence. The study shows that a mitochondrial single-locus approach can be problematic for the accurate delimitation of species, and we emphasise the need for nuclear genes as supplementary markers, when taxonomic resolution is assessed with COI barcodes.

Biodiversity of groundwater oligochaetes from a karst unit in northern Iberian Peninsula: ranking subterranean sites for conservation management

The present study suggests that the Santa Eufemia-Ereñozar karst unit in northern Iberian Peninsula is a biodiversity hotspot for groundwater oligochaetes, due to (1) the presence of a high number of stygobiotic species (corresponding to 18% of the total stygobionts known in southern Europe); (2) the comparatively high number of oligochaete species collected (corresponding to 35% of the total epigean and hypogean oligochaete species in the region); and (3) the presence of 5 species endemic to the region. A list of the oligochaete taxa found in the karst unit is presented and a conservation ranking of the cavities in the karst is proposed based on the application of four biodiversity indices (Species richness, Rarity, Vulnerability and Complementarity) to the oligochaete taxa. Vulnerability was evaluated for the first time for groundwater oligochaete taxa and it provided a useful tool to assess the protection status of oligochaetes in karstic systems. Groundwater conservation management strategies could incorporate biodiversity data from the present study.

Phylogenetic analysis of oligochaete Tubificinae (Annelida : Clitellata) based on mitochondrial sequence data

Partial sequences of the mitochondrial genes 16S rRNA and cytochrome c oxidase subunit I (COI) were used (1) to resolve the internal relationships of the subfamily Tubificinae (Annelida : Clitellata) and (2) to test the existence of cryptic species within the stygobiont oligochaete Troglodrilus galarzai (Giani & Rodriguez, 1988). Phylogenies were estimated using maximum likelihood, Bayesian inference and parsimony. Although trees were incompletely resolved at intergeneric level, a close relationship between Heterochaeta Claparede, 1863 and Troglodrilus Juget, des Châtelliers & Rodriguez was revealed, and the separation of Lophochaeta ignota Stolc, 1886 and Heterochaeta costata Claparede, 1863 from Tubifex Lamarck, 1816 was corroborated by mitochondrial molecular data. Maximum genetic divergence between allopatric populations of T. galarzai was 18% for COI (uncorrected pairwise distance), suggesting cryptic speciation within this nominal species.