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The Oxynoemacheilus bergianus species group is revised based on tree topology (ML, NJ, MP), distance (K2P and ASAP) and Poisson tree process analyses of DNA barcode data tested against morphometric and morphological characters including colour patterns. The O. bergianus species group is distinguished from other Oxynoemacheilus groups based on morphological characters: its constituent species have a slender caudal peduncle, a suborbital flap in the male, a mottled or blotched colour pattern, and lack bold, black spots on the caudal-fin base. It is also supported as a monophyletic unit in our molecular analysis. The O. bergianus group includes 10 molecular clades following congruently well-supported NJ, MP and ML based entities. Species described as O. bergianus, O. banarescui, O. erdali, O. fatsaensis, O. samanticus, and O. simavicus from Turkey, O. lenkoranensis from Azerbaijan, and O. longipinnis and O. parvinae from Iran belong to this species group. The group includes also four unnamed molecular clades. We were unable to detect external differences between any of the molecular clades in colour pattern or any morphometric or morphological characters examined. In the 10 molecular clades in the O. bergianus species group, the intraclade K2P distance ranges from 0.0–1.8% while the distances between molecular clades ranges from 0.6–5.9%. To resolve the species diversity of this group, we also analysed the intraspecific and interspecific variability in the K2P distance of DNA barcode data from 53 other Oxynoemacheilus species. Here, the intraspecific variability ranges from 0.0–2.4% while the interspecific K2P distance ranges from 1.2–20.8%. In the O. bergianus species group, only four groups are detected by the mPTP species delimitation approach distinguished by a K2P distance of 2.9% or more. We treat these four groups as valid species, corresponding to O. banarescui, O. bergianus, O. fatsaensis, and O. simavicus. Oxynoemacheilus samanticus from the Kızılırmak and Seyhan drainages, O. lenkoranensis from the Caspian basin, O. erdali from the Euphrates, and O. longipinnis and O. parvinae from the Tigris drainage are treated as synonyms of O. bergianus. Fishes from an unnamed molecular clade from the upper Tigris, and from a second unnamed clade from the upper Euphrates, are both identified as O. bergianus. Oxynoemacheilus bergianus might be a junior synonym of O. bergi from the Kura. The distribution range of O. simavicus, described from the Simav drainage in the Marmara basin, is expanded to the east and two molecularly differentiated population groups occur in the Sakarya drainage, the Büyük Melen River and potentially in other adjacent coastal streams. Oxynoemacheilus fatsaensis, described from the coastal stream Elekçi in northern Anatolia, is also widespread in the Yeşilırmak drainage. Morphological characters proposed to distinguish O. fatsaensis from the other species of the O. bergianus group could not be confirmed by our data on fishes from the Yeşilırmak. This study also discusses the theoretical background, our reasons for conducting this revision in the way we did, and what the alternatives would be.

This study summarizes results of a DNA barcoding campaign on German Diptera, involving analysis of 45,040 specimens. The resultant DNA barcode library includes records for 2,453 named species comprising a total of 5,200 barcode index numbers (BINs), including 2,700 COI haplotype clusters without species‐level assignment, so called “dark taxa.” Overall, 88 out of 117 families (75%) recorded from Germany were covered, representing more than 50% of the 9,544 known species of German Diptera. Until now, most of these families, especially the most diverse, have been taxonomically inaccessible. By contrast, within a few years this study provided an intermediate taxonomic system for half of the German Dipteran fauna, which will provide a useful foundation for subsequent detailed, integrative taxonomic studies. Using DNA extracts derived from bulk collections made by Malaise traps, we further demonstrate that species delineation using BINs and operational taxonomic units (OTUs) constitutes an effective method for biodiversity studies using DNA metabarcoding. As the reference libraries continue to grow, and gaps in the species catalogue are filled, BIN lists assembled by metabarcoding will provide greater taxonomic resolution. The present study has three main goals: (a) to provide a DNA barcode library for 5,200 BINs of Diptera; (b) to demonstrate, based on the example of bulk extractions from a Malaise trap experiment, that DNA barcode clusters, labelled with globally unique identifiers (such as OTUs and/or BINs), provide a pragmatic, accurate solution to the “taxonomic impediment”; and (c) to demonstrate that interim names based on BINs and OTUs obtained through metabarcoding provide an effective method for studies on species‐rich groups that are usually neglected in biodiversity research projects because of their unresolved taxonomy.

The Glyptothorax species inhabiting the Euphrates and Tigris drainages are reviewed and six species are recognised, one of which is described herein as new species. Glyptothorax armeniacus is endemic to headwater streams in the Euphrates drainage. Glyptothorax kurdistanicus is endemic to the upper Tigris downstream to the Lesser Zab drainage. Glyptothorax cous and G. steindachneri are riverine species widespread in both the Euphrates and Tigris drainages. Glyptothorax silviae is endemic to Iran. Glyptothorax daemon, new species, from the Greater Zab and Yanarsu in the upper Tigris drainage, is distinguished by having the thoracic adhesive apparatus strongly elevated, 1.1–1.2 times longer than wide, without tubercles on the head, well developed anteromedial striae, the medial pit without striae, and a short adipose fin. Glyptothorax daemon is separated into two mitochondrial lineages, externally indistinguishable and separated by a minimum K2P distance of 2.0% in the DNA barcode region. These lineages are paraphyletic in our analysis indicating past introgressive hybridisation with G. cous. All six species are diagnosed and all, except unstudied G. steindachneri, form distinct mitochondrial clades with between 1.2% and 3.4% minimum K2P distance between them. Species from the Euphrates and Tigris form a monophyletic mitochondrial group separated from 53 other Glyptothorax species studied from India and areas further east.

Oxynoemacheilus phasicus, new species, is described from the Rioni and Enguri River drainages in Georgia. It is distinguished from other Oxynoemacheilus species in the O. brandtii group by a mottled or marbled flank pattern in adults, a slightly emarginate caudal fin, and a deep caudal peduncle. Molecular data suggest that the new species is characterized by a minimum K2P distance of 7.5% from O. brandtii from the Kura drainage in the mtDNA COI barcode region. Oxynoemacheilus brandtii and O. elsae are re-diagnosed. A very slender Oxynoemacheilus from the Aras drainage clusters as sister to O. elsae in our molecular analysis and not with O. brandtii from the Kura River. However, it is identified as O. brandtii as it is indistinguishable from this species in morphological characters.

Oxynoemacheilus shehabi, new species, is described from the upper Orontes in southern Syria. It is distinguished from other Oxynoemacheilus species in the Levant by possession of a complete lateral line, a deeply emarginate caudal fin, a narrow caudal peduncle, a suborbital groove in male individuals, a well-developed pelvic axillary lobe, and 5–7 dark-brown bars on the flank. Molecular data suggest that the new species is characterised by a minimum K2P distance of 3.8% to O. “seyhanicola” in the COI mtDNA barcode region.

The nemacheilid genus Seminemacheilus is revised. Six species are recognised, three of them described herein as new. All species are endemic to Central Anatolia. Seminemacheilus lendlii from the Sakarya River drainage and the endorheic Lake Aksehir and Eber basins, S. ispartensis from Lake Eğirdir basin and S. ahmeti from Sultan Sazlığı are valid species. Although Seminemacheilus lendlii and S. ispartensis have almost identical COI sequences, they are distinguished by the shape of the caudal peduncle and the presence of scales on the caudal peduncle in S. ispartensis (vs. absent in S. lendlii). Seminemacheilus attalicus, new species, from Kırkgöz drainage, is distinguished by having a slightly emarginated caudal fin, a central pore in the supratemporal canal and a marbled flank pattern. Seminemacheilus ekmekciae, new species, from Lake Tuz basin is distinguished by having a roundish caudal fin, a short post-dorsal length and large brown blotches fused into stripes on the flank. Seminemacheilus tubae, new species, from Lake Beyşehir basin, is distinguished by having a truncate caudal fin and 2–5 (6) supraorbital head pores. Seminemacheilus dursunavsari from the Göksu River drainage is not a valid name and this population is identified as S. tubae.

Although aquatic macroinvertebrates and freshwater fishes are important indicators for freshwater quality assessments, the morphological identification to species-level is often impossible and thus especially in many invertebrate taxa not mandatory during Water Framework Directive monitoring, a pragmatism that potentially leads to information loss. Here, we focus on the freshwater fauna of the River Sieg (Germany) to test congruence and additional value in taxa detection and taxonomic resolution of DNA barcoding vs. morphology-based identification in monitoring routines. Prior generated morphological identifications of juvenile fishes and aquatic macroinvertebrates were directly compared to species assignments using the identification engine of the Barcode of Life Data System. In 18% of the invertebrates morphology allowed only assignments to higher systematic entities, but DNA barcoding lead to species-level assignment. Dissimilarities between the two approaches occurred in 7% of the invertebrates and in 1% of the fishes. The 18 fish species were assigned to 20 molecular barcode index numbers, the 104 aquatic invertebrate taxa to 113 molecular entities. Although the cost-benefit analysis of both methods showed that DNA barcoding is still more expensive (5.30-8.60€ per sample) and time consuming (12.5h), the results emphasize the potential to increase taxonomic resolution and gain a more complete profile of biodiversity, especially in invertebrates. The provided reference DNA barcodes help building the foundation for metabarcoding approaches, which provide faster sample processing and more cost-efficient ecological status determination.

There are particular challenges in defining the taxonomic status of recently radiated groups due to the low level of phylogenetic signal. Members of the Salmo trutta species-complex, which mostly evolved during and following the Pleistocene, show high morphological and ecological diversity that, along with their very wide geographic distribution, have led to morphological description of 47 extant nominal species. However, many of these species have not been supported by previous phylogenetic studies, which could be partly due to lack of significant genetic differences among them, the limited resolution offered by molecular methods previously used, as well as the often local scale of these studies. The development of next-generation sequencing (NGS) and related analytical tools have enhanced our ability to address such challenging questions. In this study, Genotyping-by-Sequencing (GBS) of 15,169 filtered SNPs and mitochondrial DNA (mtDNA) D-loop sequences were combined to assess the phylogenetic relationships among 166 brown trouts representing 21 described species and three undescribed groups collected from 84 localities throughout their natural distribution in Europe, west Asia, and North Africa. The data were analysed using different clustering algorithms (admixture analysis and discriminant analysis of principal components-DAPC), a Bayes Factor Delimitation (BFD) test, species tree reconstruction, gene flow tests (three- and four-population tests), and Rogue taxa identification tests. Genomic contributions of the Atlantic lineage brown trout were found in all major sea basins excluding the North African and Aral Sea basins, suggesting introgressive hybridization of native brown trouts driven by stocking using strains of the Atlantic lineage. After removing the phylogenetic noise caused by the Atlantic brown trout, admixture clusters and DAPC clustering based on GBS data, respectively, resolved 11 and 13 clusters among the previously described brown trout species, which were also supported by BFD test results. Our results suggest that natural hybridization between different brown trout lineages has probably played an important role in the origin of several of the putative species, including S. marmoratus, S. carpio, S. farioides, S. pellegrini, S. caspius (in the Kura River drainage) and Salmo sp. in the Danube River basin. Overall, our results support a multi-species taxonomy for brown trouts. They also resolve some species in the Adriatic-Mediterranean and Black Sea drainages as members of very closely related genomic clusters that may need taxonomic revision. However, any final conclusions pertaining to the taxonomy of the brown trout complex should be based on an integrative approach combining genomic, morphological, and ecological data. To avoid challenges in taxonomy and conservation of species complexes like brown trouts, it is suggested to describe species based on genomic clusters of populations instead of describing species based only on morphologically differentiated single type populations.

Pseudophoxinus cilicicus, new species, is described from the Arsuz, Ceyhan and Seyhan river drainages in the Gulf of İskenderun. It is distinguished from other members of the Pseudophoxinus zeregi species group by having a complete lateral line with 38–45 + 2–3 scales, the lower lip usually slightly projecting beyond the tip of the upper lip, a prominent black stripe along the flank, and no black pigments below the lateral line. Pseudophoxinus cilicicus is distinguished from P. zekayi by a minimum K2P distance of 3.8% based on the mitochondrial DNA barcode region. Pseudophoxinus atropatenus and P. sojuchbulagi are returned to the genus Rutilus.

The Oxynoemacheilus tigris species group is reviewed, resulting in the recognition of six species, of which two are described herein as new. Oxynoemacheilus tigris is known from the endorheic Qweik River and the Merziman River, which is a tributary of the western Euphrates. Oxynoemacheilus ercisianus is endemic to the endorheic Lake Van basin and O. hazarensis is endemic to Lake Hazar basin in the upper Tigris drainage. Oxynoemacheilus kaynaki is widespread in the Euphrates drainage. The two undescribed species occur in the Euphrates drainage. Oxynoemacheilus arsaniasus, new species, from the Murat River and the upper Karasu (Muş) River drainage, is distinguished from other species of the O. tigris group by having a bold, black, irregularly-shaped bar at the caudal-fin base, an incomplete lateral line and a scaleless body. Oxynoemacheilus muefiti, new species, from the upper Murat River drainage and a tributary to the Atatürk reservoir, is most similar to O. ercisianus, from which it is distinguished by a more slender body and a shallower dorsal adipose crest. According to our molecular data, the Qweik population of O. tigris is suspected to be introgressed by O. namiri from the Orontes drainage. Oxynoemacheilus erdali is identified as a synonym of O. bergianus as we were unable to find differences between the two species.

Article 16.4. of the International Code for Zoological Nomenclature (ICZN, 1999) requires that the fixation of name-bearing types for a new species to be explicit: “Every new specific and subspecific name published after 1999, except a new replacement name…, must be accompanied in the original publication 16.4.1. by the explicit fixation of a holotype,…..and 16.4.2. where the holotype or syntypes are extant specimens, by a statement of intent that they will be (or are) deposited in a collection and a statement indicating the name and location of that collection.” That means that for species described after 1999, the holotype must be finally deposited in a collection and it is obligatory to indicate the name of the collection and where it is located.

Paracobitis abrishamchiani, new species, is described from the southern Caspian Sea basin, where it is found in the Babol, Haraz, Siah, Tajan, Talar and Kashpal River drainages. It is distinguished from P. hircanica and P. atrakensis, the two other Paracobitis species known from the Caspian Sea basin, by having scales on the flank posterior to the dorsal-fin origin and from its congeners in the Middle East by the position of its dorsal-fin origin, having a roundish posterior narial opening, and a colour pattern consisting of many widely spaced brown spots and blotches. It is further characterized by six diagnostic nucleotide substitutions and a minimum K2P distance of 4.3% to P. persa and 4.6% to P. malapterura in the mtDNA COI barcode region.

Metabarcoding is a powerful, increasingly popular tool for biodiversity assessment, but it still suffers from some drawbacks (specimen destruction, separation, and size sorting). In the present study, we tested a non-destructive protocol that excludes any sample sorting, where the ethanol used for sample preserving is filtered and DNA is extracted from the filter for subsequent DNA metabarcoding. When tested on macroinvertebrate mock communities, the method was widely successful but was unable to reliably detect mollusc taxa. Three different protocols (no treatment, shaking, and freezing) were successfully applied to increase DNA release to the fixative. The protocols resulted in similar success in taxa detection (6.8–7 taxa) but differences in read numbers assigned to taxa of interest (33.8%–93.7%). In comparison to conventional bulk sample metabarcoding of environmental samples, taxa with pronounced exoskeleton and small-bodied taxa were especially underrepresented in ethanol samples. For EPT (Ephemeroptera, Plecoptera, Trichoptera) taxa, which are important for determining stream ecological status, the methods detected 46 OTUs in common, with only 4 unique to the ethanol samples and 10 to the bulk samples. These results indicate that fixative-based metabarcoding is a non-destructive, time-saving alternative for biodiversity assessments focussing on taxa used for ecological status determination. However, for a comprehensive assessment on total invertebrate biodiversity, the method may not be sufficient, and conventional bulk sample metabarcoding should be applied.

Biodiversity loss is mainly driven by human activity. While concern grows over the fate of hot spots of biodiversity, contemporary species losses still prevail in industrialized nations. Therefore, strategies were formulated to halt or reverse the loss, driven by evidence for its value for ecosystem services. Maintenance of the latter through conservation depends on correctly identified species. To this aim, the German Federal Ministry of Education and Research is funding the GBOL project, a consortium of natural history collections, botanic gardens, and universities working on a barcode reference database for the country’s fauna and flora. Several noticeable findings could be useful for future campaigns: (i) validating taxon lists to serve as a taxonomic backbone is time-consuming, but without alternative; (ii) offering financial incentives to taxonomic experts, often citizen scientists, is indispensable; (iii) completion of the libraries for widespread species enables analyses of environmental samples, but the process may not hold pace with technological advancements; (iv) discoveries of new species are among the best stories for the media; (v) a commitment to common data standards and repositories is needed, as well as transboundary cooperation between nations; (vi) after validation, all data should be published online via the BOLD to make them searchable for external users and to allow cross-checking with data from other countries.

The diversity of Cobitis in the Middle East is reviewed, resulting in the recognition of 30 species, of which eight are described herein as new. Two species, C. amphilekta and C. kellei , seem to be extinct. Hypotheses on species-level diversity derived from distance and Poisson tree process analyses of DNA barcode data are tested against morphometric and morphological characters including colour patterns. For species pairs separated by small K2P distances in COI sequence data we follow a practitioner-oriented diagnostic species concept, in which we recognise species only if differentiated morphologically (including by colour pattern). For all 30 species we provide diagnoses and identification keys. Cobitis afifeae , new species, from the Buyuk Menderes River drainage in the Aegean Sea basin, is distinguished by having two laminae circularis in the male, a row of blotches below Z4, a small, roundish or comma-shaped black spot at the upper caudal-fin base, and elevated mental lobes. Cobitis aliyeae , new species, from the lower Seyhan and Ceyhan River drainages, is distinguished by having two laminae circularis in the male, the blotches in Z2 and Z4 anterior to the dorsal-fin origin usually well separated from each other, and the pigmentation in Z1 well distinguished from the pigmentation in Z2. Cobiti s anabelae , new species, from the lower Orontes River drainage, is distinguished by having two laminae circularis in the male, the pigmentation in Z2 formed by small, brown spots, always much smaller than blotches in Z3, much smaller than the pupil diameter, Z2 and Z3 well separated, and no pigmentation below Z4. Cobitis erkakanae , new species, from the Golbasi Lakes, adjacent to the Ceyhan River drainage, is distinguished by having two laminae circularis in the male, no blotches below Z4, the blotches in Z2 and Z4 being horizontally elongated and often fused with adjacent ones, and the caudal fin with 4 – 6 wide, regularly-shaped, brown bands . Cobitis emrei , new species, from the Lake Sapanca basin is distinguished by having one lamina circularis in the male, a large black spot at the upper caudal-fin base, and Z3 fully covered by very small spots forming a sand-like pattern. Cobitis joergbohleni , new species, from the Sultan marshes in Central Anatolia is distinguished by having two laminae circularis in the male, and the flank colour pattern being completely disorganised, not following the Gambetta zones. Cobitis pirii, new species, from the endorheic Lake Egirdir basin and the Mediterranean Aksu and Kopru Rivers, is distinguished by having two laminae circularis in the male, a simple external part of the suborbital spine and two distinct rows of small blotches in Z4, one along the lateral midline and one distinctly below. Cobitis troasensis, new species, from the Tuzla River drainage, is distinguished by having one lamina circularis in the male and 25–36 small, comma-shaped brown blotches in Z4. A lectotype is designated for Cobitis battalgilae . As First Revisers, priority is given to Cobitis fahireae over C. kurui . Cobitis damlae and C. kurui are treated as synonyms of C. fahireae . Cobitis strumicae and C. taenia are recorded for the first time from Anatolia and C. saniae is newly documented from the Black Sea basin in Georgia. The Poisson tree process analysis of COI data proposed 31 groups, most of which could be distinguished by morphological characters. Cobitis troasensis is described based on morphological data alone .

Barbus anatolicus, new species, is described from the Kızılırmak and Yeşilırmak River drainages in the southern Black Sea basin. It is distinguished from other Barbus species in the Middle East by having 58–71 total lateral line scales, a moderately ossified last simple dorsal-fin ray, serrated along about 70–80% of its posterior margin, many small irregular shaped black or brown spots, smaller or as large as scales, often forming large, dark-brown blotches on the head, back and flank in adults and juveniles, and a concave posterior dorsal-fin margin. In addition, DNA barcode data reject the hypothesis that it belongs to one of the other species of the B. barbus species group. Barbus bergi from Bulgaria and adjacent Turkey is treated as synonym of B. tauricus. Barbus tauricus was previously believed to be restricted to the Crimean Peninsula but is found to be widespread in the Black Sea basin.

Alburnus goekhani, new species, is described from the Yeşilırmak and Kızılırmak River drainages. It belongs to the A. alburnus species group and is distinguished from other species in this group in Anatolia by having 48–56 + 2–3 scales in the lateral-line, 12–15 gill rakers, a distinct dark lateral stripe along the flank on live and preserved specimens and the anal-fin origin situated below the branched dorsal-fin ray 6–8. Alburnus goekhani is also well distinguished from other Alburnus species by its DNA barcode sequence.

The generic position of Leuciscus kurui Bogutskaya, 1995 is reviewed through a comparison of morphological and molecular characters (COI). The molecular data place L. kurui in Alburnus , close to Alburnus timarensis from the Lake Van basin. Alburnus kurui (Bogutskaya) is distinguished from this species by lacking a ventral keel and possessing both a very low number of gill rakers and midlateral scales. Alburnus selcuklui , from the upper Tigris drainage, cannot be distinguished from the widespread A. sellal and is therefore treated as a synonym of this species. Alburnus kurui Mangit & Yerli, 2018 is a junior secondary homonym of A. kurui (Bogutskaya, 1995) and A. carianorum is proposed as its replacement name. Several specimens of Alburnus caeruleus and Alburnus heckeli shared the same haplotypes as some A. sellal and therefore these species cannot always be distinguished by mitochondrial molecular characters. Alburnus caeruleus and A. heckeli are treated as valid species. Other individuals of A. caeruleus have haplotypes very different from A. sellal , and A. heckeli is well distinguished from A. sellal by having more gill rakers. The Lake Van basin as a separate freshwater ecoregion and the treatment of several species of Alburnus in synonymy of A. mento are discussed.

Oxynoemacheilus zarzianus, new species, is described from the Lesser Zab River drainage, a tributary of the lower Tigris. It is distinguished from other Oxynoemacheilus species in the Tigris drainage by having a slightly emarginate caudal fin, no suborbital groove in males, a complete lateral line, the posterior process of the bony air-bladder capsule directed posteriorly, the flank and posterior part of back covered by scales, short barbels and a deep caudal peduncle. It is the fourth Oxynoemacheilus species known from the Lesser Zab drainage, where such loaches seem to be highly isolated in headwaters. Oxynoemacheilus species diversity in the Euphrates and Tigris drainage is exceptional high. Today 22 species are known from the entire Euphrates and Tigris drainage and 15 from the Tigris drainage alone.

In order to better understand the complex geologic history of the Mediterranean area, we have analysed evolutionary history, phylogeographic structure and molecular diversity of freshwater fishes belonging to the genus Telestes. As primary freshwater fishes distributed largely in the Mediterranean basin, this genus represents a suitable model system for investigating the historical biogeography of freshwater drainage systems in southern Europe. In this investigation we have included samples representing all Telestes species and based our analyses on one mitochondrial and one nuclear gene. We have investigated phylogenetic structure inside the genus Telestes, estimated divergence times, reconstructed ancestral distribution ranges and described intraspecific molecular diversity. Diversification of Telestes started in the Early Miocene, when the ancestors of T. souffia, lineage comprising T. croaticus and T. fontinalis, and the one comprising T. pleurobipunctatus and T. beoticus got isolated. The remaining species are genetically more closely related and form a common cluster in the recovered phylogenetic trees. Complex geological history of southern Europe, including formation of continental bridges, fragmentation of landmass, closing of the sea corridor, local tectonic activities, led to complicated biogeographical pattern of this genus, caused by multiple colonization events and passovers between ancient rivers and water basins. Especially pronounced diversity of Telestes found in the Adriatic watershed in Croatia and Bosnia and Herzegovina is a consequence of a triple colonization of this area by different lineages, which led to an existence of genetically distinct species in neighboring areas. Significant intraspecific structuring is present in T. souffia, T. muticellus, T. croaticus and T. pleurobipunctatus. Besides in well-structured species, elevated levels of genetic polymorphism were found inside T. turskyi and T. ukliva, as a consequence of their old origin and unconstrained evolutionary history., This work was supported by the Czech Science Foundation (project GA15-19382S). IB received support from the SYNTHESYS Programme project CZ-TAF-4228 at the National Museum Prague, financed by European Community Research Infrastructure Action under the FP7 "Capacities" Programme. RS also received support from the SYNTHESYS Programme project ES-TAF-1187 at the Museo Nacional de Ciencias Naturales, Madrid, financed by European Community Research Infrastructure Action under the FP6 "Structuring the European Research Area" Programme.

This study represents the first comprehensive molecular assessment of freshwater fishes and lampreys from Germany. We analysed COI sequences for almost 80% of the species mentioned in the current German Red List. In total, 1056 DNA barcodes belonging to 92 species from all major drainages were used to (i) build a reliable DNA barcode reference library, (ii) test for phylogeographic patterns, (iii) check for the presence of barcode gaps between species and (iv) evaluate the performance of the barcode index number (BIN) system, available on the Barcode of Life Data Systems. For over 78% of all analysed species, DNA barcodes are a reliable means for identification, indicated by the presence of barcode gaps. An overlap between intra- and interspecific genetic distances was present in 19 species, six of which belong to the genus Coregonus. The Neighbour-Joining phenogram showed 60 nonoverlapping species clusters and three singleton species, which were related to 63 separate BIN numbers. Furthermore, Barbatula barbatula, Leucaspius delineatus, Phoxinus phoxinus and Squalius cephalus exhibited remarkable levels of cryptic diversity. In contrast, 11 clusters showed haplotype sharing, or low levels of divergence between species, hindering reliable identification. The analysis of our barcode library together with public data resulted in 89 BINs, of which 56% showed taxonomic conflicts. Most of these conflicts were caused by the use of synonymies, inadequate taxonomy or misidentifications. Moreover, our study increased the number of potential alien species in Germany from 14 to 21 and is therefore a valuable groundwork for further faunistic investigations.

Eidinemacheilus proudlovei, new species, is described from subterranean waters in the Little Zab River drainage in Iraqi Kurdistan. After the discovery of E. smithi in 1976, E. proudlovei is the second troglomorphic nemacheilid loach found in the Middle East and the second species placed in Eidinemacheilus. Eidinemacheilus proudlovei is distinguished from E. smithi by having 8+8 or 8+7 branched caudal-fin rays, no adipose keel on the caudal peduncle, enlarged jaws and a fully developed head canal system. It furthers differs substantially in its DNA barcode (gt;8% K2P distance) from all other nemacheilid loaches in the Middle East, Europe and Western India.

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No. of bitstreams: 0 Previous issue date: 2016-11-23 Villanova Univ, Dept Biol, Villanova, PA 19085 USA Museu Nacl Hist Nat & Ciencia, Lisbon, Portugal Univ Brasilia, Dept Zool, Inst Ciencias Biol, BR-70910900 Brasilia, DF, Brazil Smithsonian Inst, Natl Museum Nat Hist, Washington, DC 20560 USA Sorbonne Univ, Museum Natl Hist Nat, ISYEB, Paris, France Univ Nacl Tucuman, San Miguel De Tucuman, Argentina King Saud Univ, Riyadh, Saudi Arabia Cornell Univ, Ithaca, NY USA Univ Fed Sao Paulo, Diadema, Brazil Upper Austrian Museum, Ctr Biol, Linz, Austria Villanova Univ, Villanova, PA 19085 USA Univ Salzburg, Salzburg, Austria Plazi, Bern, Switzerland Univ Brasilia, Brasilia, DF, Brazil Univ Sao Paulo, Ribeirao Preto, Brazil Zool Forsch Museum A Koenig, Bonn, Germany Museu Paraense Emilio Goeldi, Belem, Para, Brazil Russian Acad Sci, St Petersburg, Russia Pontificia Univ Catolica Rio Grande do Sul, Porto Alegre, RS, Brazil Museo Reg Sci Nat, Turin, Italy Univ Fed Rio Grande do Sul, Porto Alegre, RS, Brazil Royal Museum Cent Africa, Tervuren, Belgium Univ Nacl Autonoma Mexico, Mexico City, DF, Mexico Univ Innsbruck, Inst Ecol, Innsbruck, Austria Hasselt Univ, Hasselt, Belgium Univ Fed Pernambuco, Recife, PE, Brazil Univ Fed Espirito Santo, Vitoria, ES, Brazil Univ Nacl Misiones, Felix De Azara, Argentina Albany Museum, Grahamstown, South Africa Natl Museum, Bloemfontein, South Africa Senckenberg Nat Hist Sammlungen, Dresden, Germany Univ Estadual Paulista, Botucatu, SP, Brazil Stephen F Austin State Univ, Nacogdoches, TX 75962 USA Landesmuseum, Leonding, Austria Univ Camerino, Camerino, Italy Univ Fed ABC, Sao Bernardo, Brazil Univ Estadual Londrina, Museu Zool, Londrina, Parana, Brazil Senckenberg Res Inst, Frankfurt, Germany Museo Civ Storia Nat, Carmagnola, Italy Polskiej Akad Nauk, Muzeum & Inst Zool, Warsaw, Poland Museo Civ Storia Nat, Genoa, Italy Russian Acad Sci, Inst Zool, St Petersburg, Russia Univ Estadual Paulista, Sao Vicente, Brazil Sorbonne Univ, ISYEB, Museum Natl Hist Nat, Paris, France Port Elizabeth Museum, Port Elizabeth, South Africa Sam Noble Museum, Norman, OK USA Harvard Univ, Museum Comparat Zool, Cambridge, MA 02138 USA North West Univ, Potchefstroom, South Africa Museu Nacl, Rio De Janeiro, Brazil Nat Hist Museum, Basel, Switzerland Senckenberg Nat Kundemuseum, Gorlitz, Germany North Carolina Museum Nat Sci, Raleigh, NC USA Univ Rennes 1, Rennes, France Univ Sao Paulo, Museu Zool, Sao Paulo, SP, Brazil Amer Museum Nat Hist, New York, NY 10024 USA Univ Huddersfield, Huddersfield, W Yorkshire, England North Dakota State Univ, Fargo, ND USA Univ Sao Paulo, Sao Paulo, SP, Brazil Fac Sci Sfax, Sfax, Tunisia DPTC PC, Manaus, Amazonas, Brazil Louisiana State Univ, Museum Nat Sci, Baton Rouge, LA 70803 USA Zool Survey India, Kolkata, India Univ Tennessee, Chattanooga, TN USA Ohio State Univ, Columbus, OH 43210 USA Univ Luterana Brasil, Museu Ciencias Nat, Canoas, Brazil Univ Firenze, Florence, Italy Univ Ovidius Constanta, Constanta, Romania Univ Mississippi, Oxford, MS USA Royal Belgian Inst Nat Sci, Brussels, Belgium Univ New Mexico, Albuquerque, NM 87131 USA Inst Nacl de Pesquisas da Amazonia, Manaus, Amazonas, Brazil Univ Stellenbosch, Matieland, South Africa Univ Fed Minas Gerais, Belo Horizonte, MG, Brazil CIBIO Ctr Invest Biodiversidade & Recursos Genet, Vairao, Portugal Museum Hist Nat, Geneva, Switzerland Univ Fed Mato Grosso, Cuiaba, Brazil Univ Austral Chile, Valdivia, Chile Univ Malaysia, Sarawak, Malaysia Indian Inst Sci, Bangalore, Karnataka, India Scottish Assoc Marine Sci, Oban, Argyll, Scotland Sam Houston State Univ, Huntsville, TX 77340 USA Museo Nacl Ciencias Nat, Madrid, Spain Sorbonne Univ, CR2P, Museum Natl Hist Nat, Paris, France Drexel Univ, Acad Nat Sci, Philadelphia, PA 19104 USA Univ Richmond, Richmond, VA 23173 USA Ctr Pesquisas Cacau, Itabuna, Brazil Univ Torino, Turin, Italy Soc Hist Nat Alcide dOrbigny, Aubiere, France Wolfden Sci Consulting, Murcia, Spain Univ Fed Rio de Janeiro, Macae, Brazil Univ Tennessee, Knoxville, TN USA Inst Humboldt, Villa De Leyva, Colombia Escuela Politec Nacl, Quito, Ecuador Calif Acad Sci, San Francisco, CA 94118 USA Museum Dept Hist Nat Var, Toulon, France Nat Environm & Wildlife Soc, Angul, India Univ Kansas, Lawrence, KS 66045 USA Kyoto Univ, Kyoto, Japan Consejo Nacl Invest Cient & Tecn, Museo Argentino Ciencias Nat Bernardino Rivadavia, Buenos Aires, DF, Argentina Univ Fed Paraiba, Joao Pessoa, Paraiba, Brazil Univ Fed Parana, Curitiba, Parana, Brazil Univ Fed Rio de Janeiro, Rio De Janeiro, Brazil Nat Hist Museum Narodini Museum, Prague, Czech Republic Nat Hist Museum Zimbabwe, Bulawayo, Zimbabwe Ateneo Manila Univ, Quezon City, Philippines Pontificia Univ Javeriana, Bogota, Colombia RACINE, St Maugan, France Univ Paris 06, Sorbonne Univ Paris, ISYEB, Paris, France Ocean Univ China, Qingdao, Peoples R China Museo Nacl Hist Nat Paraguay, San Lorenzo, Paraguay Univ Carabobo, Valencia, Venezuela Natl Ctr Biol Sci, 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Squalius namak , new species, from the endorheic Lake Namak and Kavir basins in Iran, is distinguished from the species of the genus Squalius in the Persian Gulf and the southern Caspian Sea basins by having a wide and thick symphysial knob on the lower jaw, a convex posterior anal-fin margin, a bold, dark-grey or brown, roundish or crescent-shaped blotch at the posterior tip of each flank scale and orange caudal-, anal- and pelvic-fin rays in life. Squalius namak is also characterized by four fixed, diagnostic nucleotide substitutions in the mtDNA COI barcode region.

Garra sindhi , new species, is described from Wadi Andhur in the Dhofar province of the Sultanate of Oman. It is closely related to G . dunsirei and G. smarti , two other species from the same Omani province. It differs from these species by the following combination of characters: breast covered by scales, a shallow hump on back, eyes normally developed and 8–12 gill rakers on the lower limb of the first gill arch.

Gobio nigrescens, from the Hari River drainage, is redescribed. It represents a valid species distinguished from congeners in the adjacent Caspian and Aral Sea basins by a combination of characters: a naked breast, 16 circumpeduncular scales and 6-7 irregularly shaped, black or brown blotches on back behind dorsal-fin base.

Sasanidus, new genus, is described for Noemacheilus kermanshahensis BănărescuNalbant, endemic to the Karkheh and Karun drainages in Iran. Sasanidus kermanshahensis was initially identified as a species in Oxynoemacheilus, from which it is distinguished by the absence of an external sexual dimorphism (i.e. longer pectoral fin, and nuptial tubercles on fins, head and back in males). Sasanidus is distinguished from all other genera of Nemacheilidae in the Middle East by a combination of the following character states: pelvic-fin origin behind of a vertical of the dorsal-fin origin, anus about one eye diameter in front of the anal-fin origin, dorsal adipose keel absent, a high crest on the bony capsule of the swim bladder present and colour pattern marbled or mottled or with an irregularly shaped midlateral stripe. Ilamnemacheilus longipinnis was examined and no difference could be found between Ilamnemacheilus and Oxynoemacheilus. Therefore, Ilamnemacheilus is treated as a synonym of Oxynoemacheilus. COI barcode sequences from all nemacheilid loach genera occurring in the Middle East and western India are analysed jointly for the first time. The view that Schistura is a paraphyletic assemblage is supported by the clustering of DNA sequences from 45 specimens placed in at least 20 species in the genus Schistura analysed here.

Cobitis avicennae, new species, from the Karkheh and Karun sub-drainages in the Tigris catchment is distinguished from other Cobitis species in the Persian Gulf, Kor and the southern Caspian Sea basins by having a single lamina circularis in males, a small comma-shaped black spot on the upper caudal-fin base, 5½ branched anal-fin rays, 5-6 rows of dark spots on the dorsal and caudal fins, scales below the dorsal-fin base with a small focal zone and pigmentation zone Z4 with 12-17 large, partly fused blotches. It is also distinguished from other Cobitis species in the comparison group by six fixed, diagnostic nucleotide substitutions in the mtDNA COI barcode region.

Lampetra soljani , new species, is described from the lower Neretva River in the Adriatic Sea basin. Previously it was identified as L. zanandreai . Based on morphological similarity and mitochondrial genetic data, it is related to L. lanceolata and L. ninae from the Black Sea basin. Lampetra soljani is distinguished from all other species of Lampetra by having a marmorate flank pattern in live, fully grown ammocoetes, and in some adults (vs. plain colour pattern). Lampetra soljani is distinguished from other Lampetra species by having three velar tentacles, 54–57 trunk myomeres between the last branchial opening and the anus, no posterial teeth and a bicuspid middle endolateral tooth. Lampetra soljani is also well distinguished by COI barcode data from its congeners. The new species is widespread in the Neretva River drainage and lampreys from Lake Skadar basin are likely to belong to this species also.

M. F. Geiger [et al.].- Received 14 January 2014; revision received 15 March 2014; accepted 19 March 2014, Incomplete knowledge of biodiversity remains a stumbling block for conservation planning and even occurs within globally important Biodiversity Hotspots (BH). Although technical advances have boosted the power of molecular biodiversity assessments, the link between DNA sequences and species and the analytics to discriminate entities remain crucial. Here, we present an analysis of the first DNA barcode library for the freshwater fish fauna of the Mediterranean BH (526 spp.), with virtually complete species coverage (498 spp., 98% extant species). In order to build an identification system supporting conservation, we compared species determination by taxonomists to multiple clustering analyses of DNA barcodes for 3165 specimens. The congruence of barcode clusters with morphological determination was strongly dependent on the method of cluster delineation, but was highest with the general mixed Yule-coalescent (GMYC) model-based approach (83% of all species recovered as GMYC entity). Overall, genetic morphological discontinuities suggest the existence of up to 64 previously unrecognized candidate species. We found reduced identification accuracy when using the entire DNA-barcode database, compared with analyses on databases for individual river catchments. This scale effect has important implications for barcoding assessments and suggests that fairly simple identification pipelines provide sufficient resolution in local applications. We calculated Evolutionarily Distinct and Globally Endangered scores in order to identify candidate species for conservation priority and argue that the evolutionary content of barcode data can be used to detect priority species for future IUCN assessments. We show that large-scale barcoding inventories of complex biotas are feasible and contribute directly to the evaluation of conservation priorities.

The polytypic Nicaraguan Midas cichlids (Amphilophus cf. citrinellus) have been established as a model system for studying the mechanisms of speciation and patterns of diversification in allopatry and sympatry. The species assemblage in Crater Lake Apoyo has been accepted as a textbook example for sympatric speciation. Here, we present a first comprehensive data set of population genetic (mtDNA & AFLPs) proxies of species level differentiation for a representative set of individuals of all six endemic Amphilophus species occurring in Crater Lake Apoyo. AFLP genetic differentiation was partitioned into a neutral and non-neutral component based on outlier-loci detection approaches, and patterns of species divergence were explored with Bayesian clustering methods. Substantial levels of admixture between species were detected, indicating different levels of reproductive isolation between the six species. Analysis of neutral genetic variation revealed several A. zaliosus as being introgressed by an unknown contributor, hereby rendering the sympatrically evolving L. Apoyo flock polyphyletic. This is contrasted by the mtDNA analysis delivering a clear monophyly signal with Crater Lake Apoyo private haplotypes characterising all six described species, but also demonstrating different demographic histories as inferred from pairwise mismatch distributions.