From synonym to valid species: Redescriptions of Ampharete acutifrons (Grube, 1860) and A. cirrata Webster amp; Benedict, 1887, and brief descriptions of A. baltica Eliason, 1955 and A. grubei Malmgren, 1865 (Annelida: Terebellida: Ampharetidae).

This paper aims mainly to provide clarity on the morphological characters of the type species of Ampharete, Ampharete acutifrons (Grube, 1860). Its common occurrence and wide distribution are most likely a result of misidentification of different species. Possible reasons for that are the brief original description of this species that is available solely in Latin and a series of questionable synonymizations, resulting in a confusing history of A. acutifrons. In addition to a detailed redescription of the holotype of A. acutifrons, we describe an Ampharete species from the Baltic Sea that has also been incorrectly identified as A. acutifrons for many decades. The individuals of this species agree in all diagnostic characters with those of the Northwest Atlantic species Ampharete cirrata Webster Benedict, 1887. Since no differences were found, but it is clearly distinct from A. acutifrons, A. cirrata is recognised as a valid species and consequently deleted from the synonym list of A. acutifrons. We additionally examined type material of A. grubei Malmgren, 1865 and those of A. grubei baltica Eliason, 1955, now accepted as A. baltica. According to this, both species are valid, and A. grubei must therefore also be deleted from the list of synonyms of A. acutifrons. Information on five molecular markers (Histone H3, COI, 16S, 18S, 28S) is provided for A. cirrata from the Baltic Sea. DNA sequences (H3, 16S, 28S) were identical to sequences of a specimen found in Iceland that was incorrectly determined as A. acutifrons, supporting the assumption of an amphiatlantic distribution of A. cirrata. By comparing obtained sequences to available sequences in GenBank and BOLD, we found evidence that at least four species were previously misidentified as A. acutifrons. The historical course of the taxonomy of A. acutifrons demonstrates the importance of carefully studying type material and type locality material, respectively. We believe that most previous records and synonyms of A. acutifrons have been identified incorrectly and should be re-evaluated. Additionally, an updated key to all species of Ampharete from the North Atlantic is provided.

Phylogeography of the veined squid, Loligo forbesii, in European waters.

The veined squid, Loligo forbesii Steenstrup, 1856, occurs at the European Shelf areas including the Azores and represents a valuable resource for the European commercial fishery in the North East Atlantic. However, very little is known about its population structure and phylogeography. This lack of knowledge also impedes the development of sustainable fishery management for this species. The present study combined the use of two types of markers that retrieve patterns of gene flow in different time spans; the analysis of 16 nuclear microsatellites and sequencing of the mitochondrial cytochrome oxidase subunit I (COI). Whereas the high mutation rate of microsatellites allows the description of recent patterns of connectivity in species, the lower mutation rate of COI provides phylogeographic patterns on a longer timescale. A total of 347 individuals of L. forbesii were investigated from nearly the entire distribution range of the species, including the North East Atlantic Shelf, the Azores and the Mediterranean. Individuals from the Western and Eastern Mediterranean Sea have never been included in a genetic study before. We were able to analyse COI sequences from all 12 sampling areas and define three clades of L. forbesii. Due to our large sampling area, we are presenting 13 COI-haplotypes that were previously unknown. The microsatellite analysis does not include the Azores but three main clades could be identified at the remaining 11 sampling sites. Low FST values indicate gene flow over large geographical distances. However, the genetically significant differences and an additional slight grouping in the microsatellite structure reveal that geographical barriers seem to influence the population structure and reduce gene flow. Furthermore, both markers provide strong evidence that the observed phylogeographic pattern reflects the geographical history of the Azores and the Mediterranean Sea.

Description of a new species of Sabellidae (Polychaeta, Annelida) from fresh and brackish waters in Europe, with some remarks on the branchial crown of Laonome.

In 2009, a hitherto unknown Laonome species was found in the Canal Ghent-Terneuzen in the Netherlands and subsequently in other Dutch rivers, canals and estuaries. A few years later, more unknown Laonome specimens were found in the eastern part of the Baltic Sea and in the Don River estuary, Sea of Azov. Initially, it was assumed that these specimens could represent Laonome calida Capa, 2007, originally described from Australia. In the present study we examine all these unknown European Laonome specimens and compare these specimens with the type material of L. calida from Australia. This lead to two main results: First, all specimens from Europe have the same diagnostic characters and therefore belong to one species. This finding was also supported by the results of a correspondence analysis, and genetic analyses using four different DNA sequences (COI, 16S, 28S). Second, it turned out that the type material of L. calida contains two morphologically distinct groups of specimens. The holotype and 7 paratypes are similar to each other but differ significantly from the other also similar 16 paratypes, and from all European specimens. On the basis of these observations, the Laonome specimens from European waters are described here as L. xeprovala sp. nov. We also provide the characters of the branchial crown of three Laonome species for a prospective revision of this genus.

(Cryptic) sex in the microsporidian Nosema granulosis--evidence from parasite rDNA and host mitochondrial DNA.

Microsporidia are single-celled, intracellular eukaryotes that parasitise a wide range of animals. The Nosema/Vairimorpha group includes some putative asexual species, and asexuality is proposed to have originated multiple times from sexual ancestors. Here, we studied the variation in the ribosomal DNA (rDNA) of 14 isolates of the presumed apomictic and vertically transmitted Nosema granulosis to evaluate its sexual status. The analysed DNA fragment contained a part of the small-subunit ribosomal gene (SSU) and the entire intergenic spacer (IGS). The mitochondrial cox1 gene of the host Gammarus duebeni (Crustacea) was analysed to temporally calibrate the system and to test the expectation of cophylogeny of host and parasite genealogies. Genetic variability of the SSU gene was very low within and between the isolates. In contrast, intraisolate (within a single host) variability of the IGS felt in two categories, because 12 isolates possess a very high IGS genetic diversity and two isolates were almost invariable in the IGS. This difference suggests variable models of rDNA evolution involving birth-and-death and unexpectedly concerted evolution. An alternative explanation could be a likewise unattended mixed infection of host individuals by more than one parasite strain. Despite considerable genetic divergence between associated host mitochondrial haplotypes, some N. granulosis 'IGS populations' seem not to belong to different gene pools; the relevant tests failed to show significant differences between populations. A set of recombinant IGS sequences made our data incompatible with the model of a solely maternally inherited, asexual species. In line with recent reports, our study supports the hypothesis that some assumed apomictic Microsporidia did not entirely abstain from the evolutionary advantages of sex. In addition, the presented data indicate that horizontal transmission may occur occasionally. This transmission mode could be a survival strategy of N. granulosis whose host often populates ephemeral habitats.

Into the Himalayan exile: the phylogeography of the ground beetle Ethira clade supports the Tibetan origin of forest-dwelling Himalayan species groups.

The Himalayan mountain arc is one of the hotspots of biodiversity on earth, and species diversity is expected to be especially high among insects in this region. Little is known about the origin of the Himalayan insect fauna. With respect to the fauna of high altitude cloud forests, it has generally been accepted that Himalayan lineages are derived from ancestors that immigrated from Western Asia and from adjacent mountainous regions of East and Southeast Asia (immigration hypothesis). In this study, we sought to test a Tibetan Origin as an alternative hypothesis for groups with a poor dispersal ability through a phylogeographic analysis of the Ethira clade of the genus Pterostichus. We sequenced COI mtDNA and the 18S and 28S rDNA genes in 168 Pterostichini specimens, including 46 species and subspecies of the Ethira clade. In our analysis, we were able to show that the Ethira clade is monophyletic and, thus, represents a Himalayan endemic clade, supporting endemism of two of the basal lineages to the Central Himalaya and documenting large distributional gaps within the phylogeographic structure of the Ethira clade. Furthermore, the molecular data strongly indicate very limited dispersal abilities of species and subspecies of these primary wingless ground beetles. These results are consistent with the hypothesis of a Tibetan Origin, which explains the evolution, diversity and distribution of the Himalayan ground beetle Ethira clade much more parsimoniously than the original immigration hypothesis.

The mitogenome of Gammarus duebeni (Crustacea Amphipoda): A new gene order and non-neutral sequence evolution of tandem repeats in the control region.

We determined the complete mitogenome sequence of Gammarus duebeni (Peracarida, Amphipoda). The mitogenome is circular and has a length of 15,651bp. The content corresponds to typical mitogenomes of metazoans. The gene order and transcriptional polarity of the protein-coding genes is identical to the pancrustacean ground pattern. Six tRNA genes are rearranged, making the gene order unique. Thus it will bring forward the understanding of mitogenome evolution within the Peracarida, for which much more derived gene orders are known. We postulate that the gene string trnA-trnS1 (AGN)-trnN-trnE-trnR constitutes an apomorphic character for the Amphipoda. In contrast to the relatively large genome size, we found two extremely truncated rRNA genes. The rrnL gene is the shortest (986bp) reported up until now for crustaceans. A six-time imperfect tandem repeat was observed within the control region. The inferred deterministic pattern of variation between the repeat units makes it likely, that functional constraints play an important role in the evolutionary dynamics and extant appearance of the repeat array.

Proteomic comparison of two invasive polychaete species and their naturally occurring F1-hybrids.

The mud worm genus Marenzelleria is highly invasive and is therefore studied intensively. In recently invaded habitats, sympatric populations of the sibling species Marenzelleria viridis and Marenzelleria neglecta are found. In these secondary contact zones, hybridization occurs frequently, revealing incomplete reproductive isolation between these recently diverged species. Two-dimensional polyacrylamide gel electrophoresis (2-DE) and mass spectrometric methods were applied for a comparative analysis of these species and their F(1)-hybrids. Nineteen proteins were identified by cross-species identification strategies. A low degree of interindividual variability within either species allowed characterizing qualitative species-specific differences in 2-DE spot patterns as well as in peptide maps. Species-specific peptides were found in tryptic digests of various proteins, such as glyceraldehyde-3-phosphate dehydrogenase, troponin C, gelsolin-like protein, and peroxiredoxin-1. F(1)-hybrids of M. viridis and M. neglecta showed additivity of protein spot patterns, and the presence of both parental traits was confirmed by mass spectrometric data. This study is one of few dealing with global protein expression in polychaetes and is the first proteomic description of natural F(1)-hybrids in polychaetes. It furthermore indicates the feasibility of proteomic methods for analyses of speciation in Marenzelleria siblings as well as of hybridization events in secondary contact zones in general.

Identification of Anguilla anguilla (L.) and Anguilla rostrata (Le Sueur) and their hybrids based on a diagnostic single nucleotide polymorphism in nuclear 18S rDNA.

The variability of the nuclear 18S rDNA was examined for 10 species of freshwater eels: Anguilla anguilla, A. australis, A. dieffenbachii, A. japonica, A. marmorata, A. mossambica, A. nebulosa labiata, A. obscura, A. reinhardtii and A. rostrata. We observed that a single nucleotide polymorphism (SNP) separated A. anguilla and A. japonica from the remaining taxa. While this SNP marker may be used to identify hypothetical hybrids of A. japonica and sympatric eel species, we focused on Atlantic eels to evaluate the potential for this diagnostic chromosomal marker to discriminate between individuals of A. anguilla, A. rostrata and their hybrids.

Phylogeny of Spinicaudata (Branchiopoda, Crustacea) based on three molecular markers--an Australian origin for Limnadopsis.

Taxonomy and phylogeny within the branchiopod taxon Spinicaudata are still controversial. We analyzed sequences of three gene fragments (28S rRNA, 16S rRNA and COI) from up to 41 species of the Cyzicidae, Limnadiidae and Leptestheriidae to infer their phylogenetic relationships, focusing in particular on species from Australia and their phylogenetic position within Spinicaudata. Four major monophyletic lineages could be distinguished: Limnadiidae, Leptestheriidae, Eocyzicus and all Cyzicidae except Eocyzicus. A clear genetic distinction between Australian and non-Australian Cyzicidae is well supported (i.e. Caenestheria and Caenestheriella species from Australia and Caenestheriella and Cyzicus species from Europe, Asia and North America). In the genera Eocyzicus and Eulimnadia the Australian species were closely related to those from other continents. The species of the Australian endemic genus Limnadopsis and Australian Limnadia species form a monophylum. This suggests that the origin of Limnadopsis lies in Australia and that Limnadia is not monophyletic.

Investigations on cyanobacterial diversity in a shallow estuary (Southern Baltic Sea) including genes relevant to salinity resistance and iron starvation acclimation.

The cyanobacterial diversity in the pelagic of a shallow estuary at the Southern Baltic Sea has been investigated by a combination of classical morphological data and a polymerase chain reaction (PCR)-based molecular approach. The aim of the study was to investigate possible changes in the composition of the cyanobacterial community along the salinity and nutrient gradients. For this purpose partial gene sequences of cyanobacterial 16S rDNA and of two functional genes (ggpS- salinity tolerance marker, isiA- iron starvation marker) were amplified and compared with total community DNA. Random distribution of ggpS genotypes along the salinity gradient suggests that synthesis of the osmolyte glucosylglycerol is not restricted to higher salinity sampling sites. Most of the isiA sequences formed a new homogenous cluster in a phylogenetic tree, which indicates that the indigenous cyanobacterial community comprises a group of unknown species. Minimum iron concentrations, which can activate isiA transcription in model cyanobacteria, occurred at a few sampling sites with high phytoplankton biomass and moderate salinity. Nevertheless, isiA expression could be detected at all sampling sites, which indicated restricted iron supply to cyanobacterial phytoplankton in summer.

Mutational analysis of the nf2 tumour suppressor gene in three subtypes of primary human malignant mesotheliomas.

Fourteen primary human malignant mesothelioma (HMM) samples obtained from 14 patients were screened for point mutations and microdeletions/microinsertions in exons 1-16 of the chromosome 22q-located tumour suppressor gene neurofibromin 2 (nf2) by single strand conformation polymorphism (SSCP) analysis. In one tumour (7%) a 10 basepair microdeletion of exon 10 was detected by SSCP and subsequently characterised in detail by sequencing. Deletion of the second nf2 allele in laser-microdissected regions of the 10 bp mutation-harbouring tumour was demonstrated by denaturing gradient gel electrophoresis (DGGE) analysis. Simultaneous comparative genomic hybridisation (CGH) analysis also showed losses at chromosome 22q. Our data indicate that functional loss of the NF2 protein may be involved in the formation of a subset of HMMs.