Beatriz A. Dorda

Undisclosed Taxonomic Diversity of Bathynellacea (Malacostraca: Syncarida) in the Iberian Peninsula Revealed by Molecular Data

ABSTRACT The biodiversity of Bathynellacea, a globally important group of groundwater crustacean, remains poorly known and understood. The objectives of this work were to increase the molecular information of bathynellaceans in order to test: 1) its usefulness solving taxonomic problems; and 2) evaluate the extent of cryptic speciation in a morphologically constrained clades from populations that have already been studied using only morphological methodology, contributing in this way to estimate the real diversity of Spanish subterranean fauna. We employ the COI barcode region to provide a preliminary assessment of the genetic subdivision, mtDNA lineages, of the genus Vejdovskybathynella, Bathynellidae, which has a restricted distribution to a karst system of Burgos (Spain) and was initially identified as a single species by morphological evaluation: Vejdovskybathynella edelweiss Camacho, 2007. We also studied the mtDNA lineages within six morphospecies of Parabathynellidae, five species belonging to Iberobathynella, a genus of wider distribution in the Iberian Peninsula, and one species belonging to the cosmopolitan genus Hexabathynella. The analyses of molecular data demonstrate the presence of highly divergent genetic units. We identify three divergent mtDNA clades, that may represent cryptic species that had gone unnoticed and possibly correspond to undescribed new species. We present a first preliminary molecular phylogeny of Bathynellacea, using three genera of Parabathynellidae and one genus of Bathynellidae, and one member of Anaspididae Thomson, 1893 as an out-group. The results of this study provide the first molecular data complementing the existing morphological knowledge to try to resolve the relations among Spanish genera and species of Bathynellacea through phylogenetic studies. Based on the results, we conclude that the evolutionary scenario of this special group of subterranean crustaceans cannot be revealed using only morphological information due to the presence of cryptic species.

Old and new taxonomic tools: description of a new genus and two new species of Bathynellidae from Spain with morphological and molecular characters

One new genus and two new species of the Bathynellidae Grobben, 1905 family are described from Spain. Paradoxiclamousella gen. nov. has a unique combination of morphological characters, which includes eight-segmented antenna, medial seta on exopod of antenna, antennule slightly longer than antenna, three-segmented mandibular palp, three segments on endopod of thoracopods I to V and four on thoracopods VI and VII, four spines on the sympod of uropod and three lobes (plus a very long frontal projection) on the penial region of the male thoracopod VIII. Paradoxiclamousella fideli sp. nov. displays a unique combination of morphological characters and has two exclusive characters: the shape of the outer protuberance and the bilobulated inner lobe of the male thoracopod VIII. Paradoxiclamousella pirata sp. nov. also displays a unique combination of morphological characters. Partial sequences of cytochrome c oxidase I have also been obtained from specimens of four populations, including the two type localities of the two new species. The analyses of molecular data demonstrate the presence of two highly divergent genetic units (from three localities), corresponding to two new species. The mitocondrial DNA information obtained in this study complements the traditional morphological taxonomic description supporting the validity of both new species. http://www.zoobank.org/urn:lsid:zoobank.org:pub:DD85D5EB-D02D-40FE-BEE0-3FE26D645C4E

Traditional water mite fixatives and their compatibility with later DNA studies.

This work compares frozen water, 70% alcohol, Koenikes and Angeliers fluid as preservation media for water mites in terms of their eventual facilitation of DNA extraction and amplification. The time the mites spent in the fixatives ranged between 1 week and 25 years. Two molecular markers were amplified: 16S ribosomal DNA and CoI mitochondrial DNA. DNA was extractable and could be sequenced from specimens fixed in all the above media, although this generally became more difficult as time progressed. In the light of the known characteristics of these media, the results suggest Angeliers fluid to be the most practical, especially on long expeditions.

Northernmost discovery of Bathynellacea (Syncarida: Bathynellidae) with description of a new species of Pacificabathynella from Alaska (USA)

Abstract A new species of the genus Pacificabathynella Schminke and Noodt, 1988 is described from groundwaters of Alaska (USA). This is the first record of Bathynellacea Chappuis, 1915 from the far north of America. Pacificabathynella has hitherto been known only from the states of California (one species) and Montana (three species). Pacificabathynella and Paradoxibathynella Serban, 2000 are the only genera that show sexual dimorphism in thoracopod VI. Pacificabathynella yupik sp. nov. has several unique features within the genus: antenna eight-segmented; antennule only slightly longer than the antenna; the setal formula of the maxilla (7/3/7/5); without seta on the endopod of the male thoracopod VIII; five spines on the endopod of the uropod and the endopod as long as the sympod. The new species further shows slight differences in the antennula, pars molaris of the mandible, in thoracopod I–VII and in thoracopod VIII of males and females with the other species of the genus. Sequences of mitochondrial DNA (mtDNA) cytochrome c oxidase I (COI) of five specimens of the new species were obtained to complete the morphological description with molecular data.

Iberian Peninsula and Balearic Island Bathynellacea (Crustacea, Syncarida) database.

Abstract This is the first published database of Bathynellacea. It includes all data of bathynellids (Crustacea, Bathynellacea) collected in the last 64 years (1949 to 2013) on the Iberian Peninsula and Balearic Island. The samples come from groundwater (caves, springs, wells and hyporrheic habitat associated rivers) from both sampling campaigns and occasional sampling conducted throughout the Iberian Peninsula and Balearic Islands. The dataset lists occurrence data of bathynellids distribution, sampling sites (with localities, county and geographic coordinates), taxonomic information (from family to species level) and sampling sources (collector and sampling dates) for all records. The descriptions of new species and species identifications have been carried out by an expert taxonomist (AIC) with 25 years experience in the bathynellids studies (see references). Many of the sampling sites are type localities of endemic species from Iberian Peninsula. The dataset includes 409 samples record corresponding to two families, 12 genera and 58 species, 42 of them formally described plus 16 taxa unpublished and 47 samples in study. All species known from the study area are included, which nearly sum up a quarter of species of Bathynellacea known in the world (250 species).

The collection of Bathynellacea specimens of MNCN (CSIC) Madrid: microscope slices and DNA extract

Abstract This is the first published database of a Bathynellacea Chappuis, 1915 collection of slices and DNA extracts. It includes all data of bathynellaceans (Crustacea: Syncarida) collected in the last 48 years (1968 to 2016) on the Iberian Peninsula and Balearic Islands, studied since 1984. It also includes specimens studied across many countries of Europe (Portugal, Romania, France, Italy, Slovenia, Bulgaria, and England), as well as some specimens obtained from samples of North America (Montana, Washington, Alaska and Texas), South America (Brazil, Chile and Argentina), Asia (China, Thailand, Vietnam, Mongolia and India), Africa (Morocco and Chad) and Australia (New South Wales –NSW- and Queensland). The samples come from groundwater (caves, springs, wells and hyporrheic habitat associated with rivers) obtained from both, sampling campaigns and occasional sampling efforts. The data set includes 3399 records (2657 slices and 742 DNA extracts) corresponding to three families (Parabathynellidae Noodt, 1965, Leptobathynellidae Noodt, 1965 and Bathynellidae Grobben, 1905) of the order Bathynellacea; the existence of three families is accepted, but this is a controversial issue and here is not the appropriate context to address this problem; 52 genera and 92 species formally described, in addition to 30 taxa under study and, thus, still unpublished. This represents more than half of all the genera known worldwide (80) and almost one third of the species currently known in the world (329, which increases every year). This dataset contains especially relevant collection that includes holotypes and type series of 43 new species of Bathynellacea (33 from the Parabathynellidae and ten from the Bathynellidae) described by Ana I. Camacho (AIC hereinafter); eleven of these are the type species for new genera described from all around the world, ten belonging to the Parabathynellidae and one from the Bathynellidae. As previously mentioned, these new species come from all continents, although 26 of them are from the Iberian Peninsula. The most important feature of this collection is that it has been created and reviewed by a specialist of the group (AIC), and each specimen, regardless of its shape (either permanent slices or DNA extracts), includes taxonomic, geographical and authorship information. The specialist has been involved in all stages of the process, from field sampling to the digitization of the results we are now presenting, and has worked in close collaboration with the curators responsible for the different collections involved in this project.

New Parabathynellidae species in Africa: the first bathynellids from Chad and an assay of their phylogenetic position in the order Bathynellacea (Crustacea: Malacostraca) based on 18S sequences

ABSTRACT Two new species of the genera Haplophallonella Serban and Coineau, 1975a and Racovitzaibathynella Serban and Coineau, 1994 are described from Africa. This is the first record of the members of the order Bathynellacea Chappuis, 1915 from Chad. Representatives of the genus Haplophallonella have hitherto been known only from the Ivory Coast type locality, and Racovitzaibathynella only from South Africa. Representatives of Haplophallonella irenae sp. nov. and Racovitzaibathynella dumonti sp. nov. have a unique combination of morphological characters. Haplophallonella irenae sp. nov. includes: seven-segmented antennule; five-segmented antenna; labrum with special morphology, clefted; mandible without proximal tooth; no epipodite on thoracopod I; bidentated exopod of male thoracopod VIII; triangular female thoracopod VIII; presence of basal seta on exopod of uropod and three plumose setae on endopod of uropod. Racovitzaibathynella dumonti sp. nov. includes: seven-segmented antennule; five-segmented antenna with sexual dimorphism and without seta on segment two; no epipodite on thoracopods I and II; outer lobe of male thoracopod VIII almost square, does not cover the posterior part of dentate lobe; the posterior lobe covers one part of dentate lobe, exopod with pointed distal end, basipod with distal end bilobed and endopod as two unequal smooth setae; almost triangular female thoracopod VIII and five to seven similar spines on sympod of uropod. Sequences of the 18S ribosomal RNA (rRNA) gene from 12 specimens of Racovitzaibathynella dumonti sp. nov. and two specimens of Haplophallonella irenae sp. nov. were obtained and analysed to complete the morphological description with molecular data. We have used these data to make a preliminary phylogenetic analysis with all genera of the world for those whose sequences are available (18S). Phylogenetic reconstruction of the Parabathynellidae family shows how the new African species remain clearly separated from European and Australian genus groups. http://zoobank.org/urn:lsid:zoobank.org:pub:4E7EB201-8922-403C-9546-AF8E9A409776

Description of a new genus and species of Bathynellidae (Crustacea: Bathynellacea) from Texas based on morphological and molecular characters

ABSTRACT A new genus and species of Bathynellidae is described from Texas (USA). Hobbsinella gen. nov. displays a unique combination of morphological characters including seven-segmented antenna lacking medial seta on exopod, antennule much longer than antenna, three-segmented mandibular palp with sexual dimorphism, four segments on endopod of thoracopods I–VII, five spines on sympod of uropod and three lobes on the penial region of male thoracopod VIII. Partial sequences of cytochrome oxidase I (Cox1) and 18S have been obtained from several specimens of the new species. The mitochondrial and nuclear DNA data complement the traditional morphological taxonomic description supporting the validity of the new genus. Global molecular data for the Bathynellidae demonstrate the presence of three highly divergent genetic units corresponding to three subfamilies, with the new genus confidently placed in the subfamily Gallobathynellinae. http://zoobank.org/urn:lsid:zoobank.org:pub:2FC56BDC-1B64-4F39-8B17-C041C7D61DAB