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Mediterranean freshwater systems are under threat owing to increased drought driven by climate change, intensive human land uses and non‐native species. This is causing increased fish hybridization in isolated watercourses.The genetic and morphological characteristics of hybrids of sympatric native and non‐native fish species were studied in four streams of the Mediterranean Guadalquivir basin (south‐west Spain). Fish morphology was analysed using geometric morphometrics, and molecular determination of parenthood was inferred through one mitochondrial gene (cytb) and one nuclear gene (Beta‐actin) for all hybrids and a subset of pure parental specimens.Molecular analyses confirmed hybrids between the native Squalius alburnoides and non‐native Alburnus alburnus in a stream with continuous flow. Haplotype analyses suggested that they originated from backcrossing of hybrid offspring. Intergeneric crosses between native species S. alburnoides and Pseudochondrostoma willkommii, and S. alburnoides and Iberochondrostoma lemmingii were detected in streams under reduced connectivity scenarios.Morphometrics revealed that hybrid phenotypes were similar to S. alburnoides. In some cases, molecular markers uncovered hybridization events that were neither detected in the field nor by morphometric analyses, potentially supporting a backcrossing/introgression scenario.Hybridization is likely to be increasing in Mediterranean rivers where S. alburnoides are present owing to increased fragmentation caused by summer drought exacerbated by climate change and human land uses and pressures. This can become a problem for these endemic vulnerable species if genetic diversity is lost, morphological homogenization occurs and hybrids cannot be easily detected in the field.The potential risks could be addressed by monitoring and eradication of non‐native species and segregation from natives. To avoid native–native crosses, habitat quality and desiccation risk could be tackled by improved water quality and riparian reforestation to provide shade and reduce evapotranspiration. This would need increased coordination and intervention between the institutions that share conservation responsibilities in the area. [ABSTRACT FROM AUTHOR]

Freshwater systems are among the most affected by the introduction of exotic species. The pumpkinseed sunfish Lepomis gibbosus, a centrarchid native to eastern North America, is listed among the top ten introduced freshwater fishes with the greatest ecological impact globally. Despite this, genetic and evolutionary studies of the species are still scarce. Here, we analyzed the genetic variability of introduced populations of L. gibbosus using three mitochondrial genes (COI, d-loop, and ND1). In addition, we used species distribution modeling to compare the niche of introduced versus native populations to assess the present and potential future distribution of the species under different climate change scenarios. Compared with the native populations, introduced ones present a lower level of genetic variability, indicating these populations originated from a small number of individuals from the native (Atlantic) population in the USA and Canada. The low variability was likely driven by a founder effect and subsequent bottleneck, as often occurs in invasive species. Our modeling results suggest not only that L. gibbosus modified its niche during the invasion process in Europe but also the possible global expansion of the species under future climatic conditions, which could facilitate its establishment in almost all continents. [ABSTRACT FROM AUTHOR]

Loaches of the genus Sabanejewia are model organisms for many ichthyologic studies focusing on morphological and colour variation. The bright lateral spots—a part of the mimicry of this fish—cause a taxonomic puzzle. Common practice is that morphometric and molecular studies lack habitat assessments of species originating from different environments. In this study, we analysed body shape and colour variation within two species of Sabanejewia in order to reveal whether they are more affected by phylogeny or environmental conditions. The environmental parameters were obtained using satellite mapping tools, body shape using geometric morphometry, and the molecular identification of specimens using an analysis of mitochondrial DNA (sequence of the cytochrome b gene). Generalized linear mixed models were used for the analysis of lateral spot numbers, and principal components analysis to describe the changes in landmark position defined by the permutation tests. Results show a significant impact of the locality elevation and specimen's sex on the change of the body shape. We identified significant differences in the number of lateral spots, which are considered mimicry variation across an elevation gradient. Morphological differences of Sabanejewia spp. vary with ecological factors along the river continuum. [ABSTRACT FROM AUTHOR]

We studied the population genetic structure of Cobitis vettonica, an endangered freshwater fish species endemic to the Iberian Peninsula, in order to propose a biogeographic model of the responses of species to the multiple changes that occurred in the Iberian hydrological system during the Quaternary period. We also deciphered the relationship of C. vettonica with its sister species C. paludica, particularly in sympatric areas, and provide genetic information for conservation purposes. To achieve this goal, we analyzed both mitochondrial and nuclear data (the cytochrome b and the nuclear recombination activating 1 genes) and a battery of single‐nucleotide polymorphisms (SNPs) of 248 individuals of C. vettonica or C. paludica from 38 localities, including some sympatric ones, covering the entire distribution area of C. vettonica. We highlight the important role played by the hydrogeomorphological processes and climatic changes that occurred in the Iberian Peninsula during the Quaternary on both the population structure of C. vettonica and its relationship with its sister species C. paludica. Our results support the genetic introgression of populations at the eastern limit of the distribution of C. vettonica. Furthermore, we postulate genetic introgression in sympatric areas. Finally, we propose the establishment or expansion of four Operational Conservation Units (OCUs) for C. vettonica, and highlight the threat faced by its populations due to the low level of genetic diversity detected for some of its populations and genetic introgression with C. paludica, which could eventually displace C. vettonica, resulting in a loss of diversity in this species. [ABSTRACT FROM AUTHOR]

The freshwater blenny Salaria fluviatilis has a circum-Mediterranean distribution, but it is restricted to highly specific habitats and is particularly susceptible to human disturbances, leading to the decline of Iberian populations of the species, resulting in their classification as Vulnerable. The aim of this study is to evaluate the genetic diversity and structure of the Iberian populations of S. fluviatilis to facilitate the design of robust conservation programs for the species. Potential barriers among populations and recent bottlenecks were also identified. Eleven Iberian populations (N = 337) of S. fluviatilis were genotyped for twelve polymorphic microsatellites. Our results showed strong genetic structure among Iberian populations, clustering them into four groups: north-eastern Iberian group (Fluviá, Segre, Matarraña and Cabriel rivers and Albufera Lagoon), southern Iberian group (Verde, Guadaíza and Guadalmansa rivers), Atlantic group (Zújar River and Ruidera Lakes) and Bañolas Lake (north-eastern Spain). Genetic diversity was higher in the north-eastern Iberian group (average observed heterozygosity Ho = 0.5 and number of alleles Na = 4.9) compared with in the southern Iberian group (average Ho = 0.42 and Na = 2.3), although the lowest observed heterozygosity was found in the Atlantic group (Ruidera Lakes, Ho = 0.324 ± 0.106). The analyses also revealed a recent bottleneck in the southern Iberian group. Our results not only provide essential information for future conservation plans of the Iberian populations of S. fluviatilis but also for the maintenance of the species' genetic pool in general, for which the four clusters identified in this study should be considered. [ABSTRACT FROM AUTHOR]

Accurate determination of species diversity in areas of high endemicity, particularly those lacking comprehensive systematic knowledge, represents a challenge for both taxonomists and conservationists. This need is particularly evident in areas greatly affected by anthropogenic disturbances such as the Eastern Mediterranean and its freshwater environments. To improve our knowledge of Eastern Mediterranean freshwater fishes, we phylogenetically studied Western Palearctic Cobitis species, focusing on those found in Turkey. Overall, our results provide a robust framework to assess the number of species of Cobitis. Phylogenetic reconstructions based on mitochondrial (cyt b) and nuclear (RAG1) sequences show seven major clades (Clades 1–7) grouping all Western Palearctic Cobitis species, except C. melanoleuca. In general, each major clade comprises Cobitis species that inhabit geographically close areas and have similar secondary sexual characters. Multiple divergent lineages were identified in our analyses, some of which were highly divergent such as the ones inhabiting Turkish freshwaters. Moreover, in some analyses, several of the identified lineages were incongruent with a priori defined species. Furthermore, our analyses identified eight potentially new candidate species, six that had been suggested in previous studies and two that are reported here for the first time. Our results reveal Turkey as the area with the greatest diversity of spined loaches in the Mediterranean. [ABSTRACT FROM AUTHOR]

Species of the genus Dormitator, also known as sleepers, are representatives of the amphidromous freshwater fish fauna that inhabit the tropical and subtropical coastal environments of the Americas and Western Africa. Because of the distribution of this genus, it could be hypothesized that the evolutionary patterns in this genus, including a pair of geminate species across the Central American Isthmus, could be explained by vicariance following the break-up of Gondwana. However, the evolutionary history of this group has not been evaluated. We constructed a time-scaled molecular phylogeny of Dormitator using mitochondrial (Cytochrome b) and nuclear (Rhodopsin and β-actin) DNA sequence data to infer and date the cladogenetic events that drove the diversification of the genus and to relate them to the biogeographical history of Central America. Two divergent lineages of Dormitator were recovered: one that included all of the Pacific samples and another that included all of the eastern and western Atlantic samples. In contrast to the Pacific lineage, which showed no phylogeographic structure, the Atlantic lineage was geographically structured into four clades: Cameroon, Gulf of Mexico, West Cuba and Caribbean, showing evidence of potential cryptic species. The separation of the Pacific and Atlantic lineages was estimated to have occurred ~1 million years ago (Mya), whereas the four Atlantic clades showed mean times of divergence between 0.2 and 0.4 Mya. The splitting times of Dormitator between ocean basins are similar to those estimated for other geminate species pairs with shoreline estuarine preferences, which may indicate that the common evolutionary histories of the different clades are the result of isolation events associated with the closure of the Central American Isthmus and the subsequent climatic and oceanographic changes. [ABSTRACT FROM AUTHOR]

We present a phylogenetic investigation of the Northern Clade, the major monophyletic clade within the freshwater fish family Cobitidae, one of the most prominent families of freshwater fishes found in Asian and European waters. Phylogenetic reconstructions based on the cytochrome b and RAG-1 genes show the genera Microcobitis, Sabanejewia, Koreocobitis and Kichulchoia as monophyletic groups. These reconstructions also show a Cobitis sensu lato and a Misgurnus sensu lato group. The Cobitis sensu lato group includes all species of Cobitis, Iksookimia, Niwaella and Kichulchoia, while the Misgurnus sensu lato group includes Misgurnus, Paramisgurnus and Koreocobitis. Although the monophyly of both the Cobitis sensu lato and Misgurnus sensu lato groups is supported, relationships within the groups are incongruent with current generic definitions. The absence of monophyly of most genera included in the Cobitis sensu lato group (Cobitis, Iksookimia and Niwaella) or their low genetic differentiation (Kichuchoia) supports their consideration as synonyms of Cobitis. Molecular phylogenies indicate that the Asian species of Misgurnus experienced a mitochondrial introgression from a lineage of Cobitis. We also find two nuclear haplotypes in the same Cobitis species from the Adriatic area that, in the absence of morphological differentiation, may indicate molecular introgression. Most lineages within the Northern Clade consist of species found in East Asia. However, some lineages also contain species from Europe and Asia Minor. The phylogenetic relationships presented here are consistent with previous studies suggesting an East Asian origin of the Northern Clade. According to the current distributions and phylogenetic relationships of the Misgurnus sensu lato and Cobitis clade lineages, particularly of M. fossilis and C. melanoleuca, the range expansion of East Asian species into Europe was most likely via Siberia into Northern and Central Europe. Phylogenetic analyses also show that the Cobitis sensu lato group consists of two clear subgroups (I and II), each presenting geographical differences. Subgroup I is distributed exclusively in East Asian drainages with an Eastern European offshoot (C. melanoleuca), whereas Subgroup II includes species widespread throughout Europe (including the Mediterranean), Asia Minor, the Black Sea and the Caucasus, with some lineages related to species restricted to East Asia. [ABSTRACT FROM AUTHOR]

This study represents a thorough analysis of Codoma, a monotypic genus endemic to north-western Mexico. A previous morphological analysis of the species concluded that there exists several morphological groups in Codoma ornata, suggesting diversity in Codoma could be underestimated. No studies have examined the genetic diversity in Codoma ornata to test this hypothesis and identify independent lineages. We present a phylogeographic analysis using one mitochondrial and two nuclear genes, and specimens from across nine major drainages in both the Chihuahuan Desert and the Sierra Madre Occidental of western Mexico. All genes and analyses recovered populations of Codoma in a well-supported clade and sister to Tampichthys, and this clade sister to Cyprinella. Analyses of both mitochondrial and nuclear genes indicated Codoma is not monotypic and recover more diversity in the genus than currently recognized. The four (mitochondrial) and five (nuclear) genetically distinct lineages are consistent with those groups outlined in the prior morphological study of the genus. Composition and distribution of these major lineages is also consistent with prior biogeographic hypothesis for other fishes in the region, supporting an ancestral Rio Grande system extending south towards central Mexico. Fragmentation of this paleosystem was followed by allopatric speciation in the Chihuahuan Desert. These results suggest a scenario of long-term isolation in four major regions (upper Conchos, lower Conchos, Nazas, upper Mezquital). Resolution of the diversity and biogeography of these lineages has many implications for various biological disciplines, especially for evolutionary and conservation studies. [ABSTRACT FROM AUTHOR]

We assessed the genetic variability of the Siberian spined loach Cobitis melanoleuca across its unusually broad distribution that encompasses areas greatly affected by Pleistocene glaciations. Due to extensive morphological variation among their populations, the taxonomic status of C. melanoleuca is complicated. It is unclear whether C. melanoleuca represents a single taxonomic unit or contains several species or subspecies. Our analyses showed low genetic variability in all populations without any phylogenetic structure. The absence of molecular distinctiveness indicates the conspecificity of all C. melanoleuca populations. Only a few common haplotypes shared by East Asian, Siberian and European populations were found at high frequency in the nuclear genes analysed. At the mitochondrial level, Siberian populations shared haplotypes with populations located at both extremes of the species' range suggesting central populations as a source of current mitochondrial variability. Unimodal mismatch distributions and significant values from neutrality tests support a recent expansion of C. melanoleuca. Our time estimates suggest a postglacial colonisation of European waters around 1.0 MYA, indicating that C. melanoleuca may represent the last cobitid immigrant in Europe that used the northern route across Siberia to expand its range. [ABSTRACT FROM AUTHOR]

Russian Far East loaches of the genera Cobitis and Misgurnus are among members of the family Cobitidae with poorly understood systematics. In this study we present phylogenetic hypotheses based on mitochondrial (cytochrome b) and nuclear (RAG-1) sequences. All analyses recovered comparable topological phylogenies, and all data sets supported the non-monophyly of the genera Cobitis and Misgurnus. Both genera are represented by multiple lineages that in some cases do not correspond to the species described. We found some phylogenetic incongruities for the genus Misgurnus ( M. mohoity and M. anguillicaudatus) that are explained by ancient hybridization, as was suggested previously for M. anguillicaudatus. The revealed phylogenetic relationships suggest that Paramisgurnus should be treated as a synonym of Misgurnus and M. bipartitus as a synonym of M. mohoity. All analyses recovered C. choii as a member of the genus Cobitis, confirming previous taxonomic conclusions. Most of the molecular lineages found follow currently recognized taxa with some exceptions, such as M. anguillicaudatus and C. lutheri. Phylogenetic relationships recover several unrelated lineages of M. anguillicaudatus and suggest additional studies to solve current taxonomic uncertainty. We found that C. lutheri is a non-natural group that contains two unrelated lineages: specimens of C. lutheri from the Far East of Russia collected close to the type locality and a second lineage with specimens of C. lutheri from Korea, the identification of which must be revised. The study provides evidence of the presence of the Far East species M. nikolskyi in Sakhalin Island, but simultaneously shows conspicuous genetic distinctiveness between the island and the mainland populations. [ABSTRACT FROM AUTHOR]

Geomorphic evolution of river basins can shape the structure and diversity of aquatic communities, but understanding the biological significance of basin evolution can be challenging in semi-arid regions with ephemeral or endorheic conditions and complex drainage configurations such as the Sierra Madre Occidental (SMO) in North America. In this study, we characterized range-wide patterns of genetic variation in the Mexican stoneroller ( Campostoma ornatum) to infer how orogenic and erosional influences on river basin connectivity have given rise to the diverse and largely endemic freshwater communities across the SMO region. Twelve drainage basins across northern Mexico and the south-western United States, centred on the SMO. We collected 202 specimens from 98 localities across the range of C. ornatum. We performed phylogenetic analyses of DNA sequences from one mitochondrial (cytochrome b) and one nuclear (intron S7) gene. Phylogenetic trees were estimated for each data set using maximum likelihood and Bayesian inference. Phylogenetic analyses consistently resolved a monophyletic C. ornatum composed of multiple evolutionary lineages within two markedly divergent clades that differentiate northern drainages from southern drainages in the SMO region. Within-clade patterns of divergence corresponded to fine-scale geographic structure within and among SMO drainage basins. However, the geographic distribution of evolutionary lineages within the northern and southern clades did not always correspond to the geographic configuration of drainage basins. Some subclades encompassed multiple drainages, and individuals from a single drainage were sometimes recovered in multiple subclades. Our findings indicate that a common ancestor of Mexican Campostoma is likely to have entered north-west Mexico through an ancient Rio Grande system that extended as far south as the Rio Nazas and Rio Aguanaval. The geographic orientation of the two strongly divergent clades recovered within C. ornatum provides evidence of long-standing isolation of southern basins from northern basins within the ancestral Rio Grande system, possibly due to the combined influence of tectonic events and increasing regional aridity. Geographic patterns of genetic variation also provide evidence of range expansion from Atlantic to Pacific drainages due to drainage evolution and river capture events, as well as further inter-basin exchange via more recent headwater capture events, hydrological connections and possible anthropogenic introductions. [ABSTRACT FROM AUTHOR]

The molecular divergence and phylogenetic relationships of the Iberian populations of Aphanius iberus were established using allozymes and the complete cytochrome b gene sequence. Congruent results were found with both nuclear and mitochondrial molecular markers. The Mediterranean and Atlantic populations are clearly differentiated into two independent lineages. Their high molecular divergence suggests an early isolation, and the absence of gene flow among the populations indicates their independent evolution. The nuclear and mitochondrial data reveal monophyletic clustering of the two geographical lineages, but provide weak support for the population relationships. However, the mitochondrial results differentiated the Villena population as a distinct mitochondrial unit within the Mediterranean group. Geographically broad studies across the distribution range of A. iberus have helped to elucidate the patterns of diversification of this species. The genetic divergence found between the Atlantic and the Mediterranean populations is of the same order as those found among recognized species of cyprinodontids. The identification of two discrete evolutionary lineages has important implications for the conservation of this species, since its recovery requires the recognition and preservation of natural diversity. The Mediterranean and Atlantic lineages should be managed separately to avoid loss of their genetic identity, and the genetic uniqueness of the populations should be preserved by using wild stocks as the source of genetic diversity in captive breeding programmes. [ABSTRACT FROM AUTHOR]