Novel molecular resources for single-specimen barcoding of enigmatic crustacean y-larvae.

Despite discovery more than 100years ago and documented global occurrence from shallow waters to the deep sea, the life cycle of the enigmatic crustacean y-larvae isincompletely understood and adult forms remain unknown. To date, only 2 of the 17 formally described species, all based on larval stages, have been investigated using an integrative taxonomic approach. This approach provided descriptions of the morphology of the naupliar and cyprid stages, and made use of exuvial voucher material and DNA barcodes. To improve our knowledge about the evolutionary history and ecological importance of y-larvae, we developed a novel protocol that maximises the amount of morpho-ecological and molecular data that can be harvested from single larval specimens. This includes single-specimen DNA barcoding and daily imaging of y-nauplii reared in culture dishes, mounting of the last naupliar exuviae on a slide as a reference voucher, live imaging of the y-cyprid instar that follows, and fixation, DNA extraction, amplification and sequencing of the y-cyprid specimen. Through development and testing of a suite of new primers for both nuclear and mitochondrial protein-coding and ribosomal genes, we showcase how new sequence data can be used to estimate the phylogeny of Facetotecta. We expect that our novel procedure will help to unravel the complex systematics of y-larvae and show how these fascinating larval forms have evolved. Moreover, we posit that our protocols should work on larval specimens from a diverse array of moulting marine invertebrate taxa.

Diversity, habitat endemicity and trophic ecology of the fauna of Loki's Castle vent field on the Arctic Mid-Ocean Ridge.

Loki's Castle Vent Field (LCVF, 2300 m) was discovered in 2008 and represents the first black-smoker vent field discovered on the Arctic Mid-Ocean Ridge (AMOR). However, a comprehensive faunal inventory of the LCVF has not yet been published, hindering the inclusion of the Arctic in biogeographic analyses of vent fauna. There is an urgent need to understand the diversity, spatial distribution and ecosystem function of the biological communities along the AMOR, which will inform environmental impact assesments of future deep-sea mining activities in the region. Therefore, our aim with this paper is to provide a comprehensive inventory of the fauna at LCVF and present a first insight into the food web of the vent community. The fauna of LCVF has a high degree of novelty, with five new species previously described and another ten new species awaiting formal description. Most of the new species from LCVF are either hydrothermal vent specialists or have been reported from other chemosynthesis-based ecosystems. The highest taxon richness is found in the diffuse venting areas and may be promoted by the biogenic habitat generated by the foundation species Sclerolinum contortum. The isotopic signatures of the vent community of LCVF show a clear influence of chemosynthetic primary production on the foodweb. Considering the novel and specialised fauna documented in this paper, hydrothermal vents on the AMOR should be regarded as vulnerable marine ecosystems and protective measures must therefore be implemented, especially considering the potential threat from resource exploration and exploitation activities in the near future.

Investigation of the reproductive system in calanoid copepods: A new approach using 3D reconstructions from serial semi-thin cross-sections in Calanus glacialis and Metridia longa.

The structure of the female reproductive system of the calanoid copepods Calanus glacialis and Metridia longa from the White Sea was studied using light microscopy, scanning and transmission electron microscopy, as well as confocal laser scanning microscopy. For the first time, we applied also the method of 3D reconstructions from semi-thin cross-sections to visualize the general plan of the reproductive system in both species. The application of a combination of methods provided novel and detailed information on the genital structures and muscles located in the genital double-somite (GDS) as well as structures used for the reception and storage of spermatozoa, fertilization and release of eggs. An unpaired ventral apodeme and associated muscles located in the GDS are described for the first time for calanoid copepods. The role of this structure in copepod reproduction is discussed. Stages of oogenesis and the mechanism of yolk formation in M. longa are studied using semi-thin sections for the first time. A combination of non-invasive (LM, CLSM, SEM) and invasive techniques (semi-thin sections and TEM) applied in this study substantially improves our understanding of the functioning of the genital structures in calanoid copepods and could be recommended as a standard set of methods for future research in the reproductive biology of copepods.

A synthesis of the external morphology of cypridiform larvae of Facetotecta (crustacea: Thecostraca) and the limits of the genus Hansenocaris.

Although the naupliar and cypridiform stages of the enigmatic y-larvae of Facetotecta have been found in the marine plankton worldwide, they still represent the last significant group of crustaceans for which the adult forms are still unknown. From a number of y-cyprids representing different taxa from different locations, we employ scanning electron microscopy to describe fine morphological details of all external structures of this unique larval form. We document different segmentation patterns of the abdomen and presence/absence of the labrum and structural differences in the antennules, labrum, paraocular process, thoracopods, and telson lend support for the erection of several new genera as opposed to the single Hansenocaris. The data presented here emphasize the morphological limits of the genus Hansenocaris and the "bauplan" of cyprydiform larvae of Facetotecta. Although the optimum pathway is a joint analysis of both molecular and morphological characters, we use the morphological characters of y-cyprids to align them cladistically and determine the limits of the genus Hansenocaris s.s. and describe common characters for all y-cyprids including six pairs of the lattice organs instead five pairs considered as a ground pattern for all Thecostraca. We also determine plesiomorphic and apomorphic characters of all known y-cyprids and separate them from other thecostracan cypridiform larvae.

Microdajus tchesunovi sp. n. (Tantulocarida, Microdajidae) - a new crustacean parasite of from the White Sea.

A new species of Tantulocarida, Microdajus tchesunovi sp. n., was found on tanaid host, Typhlotanais sp. of the family Nototanaidae, collected from silty sediment in the Kandalaksha Bay in the White Sea. Several tantulus larvae, developing males at different stages of metamorphosis, and early stages of parthenogenetic females were found attached to different sites of their hosts. Ultrastructure of the new species was studied with SEM. Microdajus tchesunovi sp. n. can be easily distinguished from other species of the genus Microdajus Greve by the presence of a pair of longitudinal dorsal lamellae at the anterior end of the cephalon. It is also characterized by the presence of an endopod seta on the sixth thoracopod and the absence of thoracopodal endites. A morphological comparison of species of the family Microdajidae is presented in tabular form.

Serratotantulus chertoprudae gen. et sp. n. (Crustacea, Tantulocarida, Basipodellidae): A new tantulocaridan from the abyssal depths of the Indian Ocean.

A single tantulus larva was found at the abyssal depth of the Indian Ocean attached to a harpacticoid host of the family Cletodidae. It represents a new genus and species of Tantulocarida, family Basipodellidae. Its ultrastructure was studied with SEM. This genus can be easily distinguished from the other genera of Basipodellidae by the pore pattern, bilobed oral disk with strong longitudinal ridges and the posterior projection of the cephalic shield. A morphological analysis of two related families Basipodellidae and Deothertridae shows that they represent polyphyletic taxa and need further revision.