Genetic Differentiation Among Annual Fishes of the Genus Cynolebias (Cyprinodontiformes, Rivuluidae) in a Biosphere Reserve Site from Uruguay

The annual killifishes of the genus Cynolebias include one well supported clade composed of three taxa: Cynolebias adloffi, from Porto Alegre (southern Brazil), C. cf. adloffi and Cynolebias viarius, parapatrically distributed in southeastern Uruguay. Cynolebias cf. adloffi has always been considered an intergradation complex of populations between C. adloffi and C. viarius showing high levels of morphological variability and its systematic affinities have been historically discussed. This study clarified the patterns of genetic differentiation of C. cf. adloffi by means of RAPD markers to define its taxonomic status as a possible separated taxon. Multidimensional scaling analysis reveals that C. cf. adloffi is composed of two highly structured genetic groups (northern and southern) with discrete geographical distributions. The estimate of gene diversity for RAPD markers could suggest a scenario in which the rainy season flood pool populations across large distances could make intrapopulation variation high but interpopulation variation low. Phylogenetic analyses (UPGMA and parsimony) show different and unstable patterns of relationships among the taxa analyzed. In the UPGMA tree, C. viarius and C. cf. adloffi appear as sister taxa, while parsimony analysis does not support a unique hypothesis of relationship. Our analyses are in agreement with the hypothesis that C. cf. adloffi may well be a case of ancient hybrid ancestry followed by its divergence associated with Pleistocene and post-Pleistocene environmental changes in this region.     

Pattern of differentiation in the annual killifish genus Austrolebias (Cyprinodontiformes: Rivulidae) from a biosphere reserve site in South America: a multidisciplinary approach

The annual killifish genus Austrolebias includes approximately 38 species distributed throughout the Paraná-Plata basin and Patos-Merín system. Within the Austrolebias adloffi species complex, the Uruguayan populations of Austrolebias charrua were considered as an intergradation between A. adloffi and Austrolebias viarius populations. Austrolebias charrua presents an intermediate phenotype between both taxa and high levels of morphological and chromatic variability. In the present study, we incorporate different methodological approaches (molecular, morphology, and gamete ultrastructure) to elucidate the pattern of differentiation among the parapatric taxa ( A. charrua , Austrolebias reicherti , A. viarius ) distributed in a Biosphere Reserve Site. Analyses of cytochrome b sequences show high values of DNA polymorphism, in particular for A. charrua. This is in accordance with both morphological and gametic variation. Using a statistical parsimony network based on these sequences and analysis of morphological data, past fragmentation and range expansion involving perhaps secondary contact between A. charrua and A. reicherti could be proposed. Coloration pattern and morphometric analyses showed an unexpected higher similarity between the most distantly-related taxa, A. viarius and A. charrua . This could be the result of retention of ancestral polymorphisms, especially in A. charrua populations from ponds of higher elevation, or to directional selection acting in similar ecological environments. Because these annual killifish species are considered endangered, our work reinforces the high priority need to include them in a conservation programme.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 620–635.     

Mitochondrial genes: signals and noise in the phylogenetic reconstruction of the annual killifish genus Cynolebias (Cyprinodontiformes, Rivulidae)

The genus Cynolebias (Cyprinodontiformes: Rivulidae) is a locally endemic and speciose group of Neotropical fishes with an annual life cycle. Members of the genus vary greatly in morphology and behaviour, and extensive interspecific karyotypic divergence has been documented among species from Uruguay, Argentina and Rio Grande do Sul, Brazil. We present a molecular systematic hypothesis of the relationships between these Cynolebias species based on phylogenetic analysis of a combined dataset containing 1825 base pairs of DNA sequence, representing three mitochondrial genes. The protein-coding cytochrome -b gene, the 12S and 16S rRNA mitochondrial genes, alone and in combination, yield robust support for monophyly within Cynolebias . Furthermore, our analyses identify two major Cynolebias clades, one of which contains at least four monophyletic groups. Corrected mtDNA genetic distances range from 5.2 to 17.5% between Cynolebias species, and application of a molecular clock suggests the occurrence of two pulses of cladogenesis, one in the late Miocene and another in the Pliocene–Pleistocene. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 76 , 49–59.     

Patterns of population differentiation in annual killifishes from the Paraná–Uruguay–La Plata Basin: the role of vicariance and dispersal

Aim To elucidate the role of vicariance versus dispersal at the microevolutionary scale in annual killifish populations belonging to the Austrolebias bellottii species complex (Rivulidae). Within this complex, A. bellottii and A. apaii have low vagility and occur widely within the study area, making them excellent models for testing biogeographic hypotheses of differentiation. Location South America, in the Paraná–Uruguay–La Plata river basin. Methods Molecular data and morphometric analyses were used to reconstruct the phylogeographic history and morphological variation of 24 populations of two taxa of the A. bellottii species complex. Phylogenetic analyses using maximum likelihood (ML) and Bayesian inference (BI) model‐based methods, estimates of clade divergence times implemented in beast , non‐metric multidimensional scaling, analysis of molecular variance results, and morphological analyses elucidated the role of vicariance versus dispersal hypotheses in population differentiation in the aforementioned river basin. Results In the A. bellottii species complex from the Paraná–Uruguay–La Plata river basin, past allopatric fragmentation from vicariance events seems to be the most plausible scenario for diversification since the Late Miocene and more recently since the Plio‐Pleistocene. The Plio‐Pleistocene vicariance produced the differentiation of three major clades in A. bellottii populations. One clade from the eastern Uruguay River drainage was separated from another in western Uruguay and the Paraná–La Plata River drainages. A later vicariance event split populations to the south (lower Paraná–La Plata Basin) and north (middle Paraná) of the western Paraná River drainage. However, our results do not exclude the possibility of dispersal events among A. bellottii populations from both the Uruguay and Paraná river drainages, which could occur in these river basins during hypothesized connectivity cycles of the Late Pliocene and Pleistocene. Main conclusions Past allopatric fragmentation caused by different vicariance events seems to be the main driver of diversification in the A. bellottii species complex since the Plio‐Pleistocene. However, the current molecular data suggest that populations from both drainages of the Paraná–Uruguay rivers may have experienced cycles of connectivity during the Pleistocene, perhaps including multiple vicariance or dispersal events from populations located in the western lower Uruguay River drainage, which encompassed climatic and geological changes in the Paraná–Uruguay–La Plata Basin.     

Historical biogeography of the Western Rattlesnake (Serpentes: viperidae: Crotalus viridis), inferred from mitochondrial DNA sequence information

We infer the phylogeography of the Western Rattlesnake (Crotalus viridis) using phylogenetic analysis of mitochondrial DNA sequences from 1345 bp of the genes for cytochrome b and NADH dehydrogenase subunit 4. Two main clades are revealed: one includes populations from east and south of the Rocky Mountains (conventionally referred to as Crotalus viridis viridis and C. v. nuntius), and the other consists of populations west of the Rocky Mountains. Within the western clade, a population from southern Arizona (C. v. cerberus) represents the sister taxon to the remaining western populations. The conventional subspecies recognized in this species do not fully correspond to the phylogenetic pattern, and a review of the systematic status of several populations is needed. Our data allow the inferences that small body size evolved twice and that the ability of one population (C. v. concolor) to secrete highly lethal toxins related to Mojave toxin arose within the complex. Our phylogeny should represent the basis for further studies on the causes of geographical variation in this complex.     

Phylogeography of Iurus dufoureius (Brullé, 1832) (Scorpiones, Iuridae)

Aim  This study uses molecular data in conjunction with palaeogeography to infer the most plausible biogeographical scenario accounting for the current distributional pattern of Iurus dufoureius .
Location  North-eastern Mediterranean region.
Methods  Sequencing of a 441-bp segment of the mitochondrial 16S rRNA gene in seven populations covering the whole distributional range of the species. Phylogenetic analyses performed included neighbour joining, maximum parsimony and Bayesian inference.
Results  The molecular phylogeny showed that two Iurus clades are strongly supported. These clades correspond to the two subspecies Iurus dufoureius dufoureius and Iurus dufoureius asiaticus , currently recognized within the genus. The assumption of a clock-like evolution could not be rejected, and this enabled us to estimate an approximation of the local rate of evolution for the I. dufoureius lineages. Based on the estimated evolutionary rate (0.79 ± 0.17 Myr⁻¹), the split between the two Iurus clades occurred c. 8 Ma.
Main conclusions  Contrary to what was believed in the past, the genus Iurus is an old north-eastern Mediterranean genus that has been differentiating in southern Greece and south-west Turkey at least from the middle Miocene. According to the phylogenetic trees obtained and the dating of the divergence times of lineages, the genus dispersed into the Aegean Archipelago when the Aegean was still a uniform land mass. Although the phylogenetic relationships of I. d. dufoureius populations have been shaped by the most recent vicariant events, the phylogenetic relationships of I. d. asiaticus populations are mostly attributable to older palaeoevents occurring in the area.     

Karyotype differentiation in Chromaphyosemion killifishes (Cyprinodontiformes, Nothobranchiidae): patterns, mechanisms, and evolutionary implications

Chromaphyosemion killifishes are a karyotypically highly diverse group of small, sexually dimorphic fishes living in rainforest rivulets in tropical West and Central Africa. In the present study, we used various chromosome banding and staining techniques to analyse the karyotypes of 13 populations representing seven described species ( Chromaphyosemion loennbergii , Chromaphyosemion punctulatum , Chromaphyosemion splendopleure , Chromaphyosemion volcanum , Chromaphyosemion malumbresi , Chromaphyosemion melanogaster , Chromaphyosemion bitaeniatum ) and two undescribed forms ( Chromaphyosemion cf. lugens , Chromaphyosemion sp. Rio Muni GEMHS00/41). Diploid chromosome numbers (2 n ) and the number of chromosome arms (NF) ranged from 2 n  = 24 in C. malumbresi to 2 n  = 40 in C. bitaeniatum and from NF = 40 in C. volcanum and C. cf. lugens to NF = 54 in one population of C. loennbergii . A tentative XX/XY sex chromosome system was revealed in C. loennbergii , C. melanogaster , C. malumbresi , and Chromaphyosemion sp. Rio Muni GEMHS00/41. Mapping cytogenetic data for all described Chromaphyosemion species onto a recently published mitochondrial DNA phylogeny revealed a complex pattern of chromosomal evolution with several independent reductions of 2 n and independent modifications of NF and nucleolus organizer region phenotypes. Together with the results of preliminary crossing and mate choice experiments, the cytogenetic and molecular phylogenetic data suggest that, contrary to previous hypotheses, chromosomal rearrangements are probably not the most important and certainly not the only factor driving speciation in Chromaphyosemion killifishes.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 143–153.     

Phylogeny, divergence times and species limits of spiny lizards (Sceloporus magister species group) in western North American deserts and Baja California

The broad distribution of the Sceloporus magister species group (squamata: phrynosomatidae) throughout western North America provides an appropriate model for testing biogeographical hypotheses explaining the timing and origins of diversity across mainland deserts and the Baja California Peninsula. We inferred concordant phylogenetic trees describing the higher-level relationships within the magister group using 1.6 kb of mitochondrial DNA (mtDNA) and 1.7 kb of nuclear DNA data. These data provide strong support for the parallel divergence of lineages endemic to the Baja California Peninsula (S. zosteromus and the orcutti complex) in the form of two sequential divergence events at the base of the magister group phylogeny. A relaxed phylogenetic analysis of the mtDNA data using one fossil and one biogeographical constraint provides a chronology of these divergence events and evidence that further diversification within the Baja California clades occurred simultaneously, although patterns of geographical variation and speciation between clades differ. We resolved four major phylogeographical clades within S. magister that (i) provide a novel phylogenetic placement of the Chihuahuan Desert populations sister to the Mojave Desert; (ii) illustrate a mixed history for the Colorado Plateau that includes Mojave and Sonoran Desert components; and (iii) identify an area of overlap between the Mojave and Sonoran Desert clades near Yuma, Arizona. Estimates of bidirectional migration rates among populations of S. magister using four nuclear loci support strong asymmetries in gene flow among the major mtDNA clades. Based on the nonexclusivity of mtDNA haplotypes, nuclear gene flow among populations and wide zones of phenotypic intergradation, S. magister appears to represent a single geographically variable and widespread species.     

Filling the gaps: phylogeography of the self-fertilizing Kryptolebias species (Cyprinodontiformes: Rivulidae) along South American mangroves

Mangrove killifishes of the genus Kryptolebias have been historically classified as rare because of their small size and cryptic nature. Major gaps in distribution knowledge across mangrove areas, particularly in South America, challenge the understanding of the taxonomic status, biogeographical patterns and genetic structuring of the lineages composing the self-fertilizing “Kryptolebias marmoratus species complex.” In this study, the authors combined a literature survey, fieldwork and molecular data to fill major gaps of information about the distribution of mangrove killifishes across western Atlantic mangroves. They found that selfing mangrove killifishes are ubiquitously distributed across the Caribbean, Central and South American mangroves and report 14 new locations in South America, extending the range of both the “Central clade” and “Southern clade” lineages which overlap in the Amazon. Although substantial genetic differences were found between clades, the authors also found further genetic structuring within clades, with populations in Central America, north and northeast Brazil generally showing higher levels of genetic diversity compared to the clonal ones in southeast Brazil. The authors discuss the taxonomic status and update the geographical distribution of the Central and Southern clades, as well as potential dispersal routes and biogeographical barriers influencing the distribution of the selfing mangrove killifishes in the western Atlantic mangroves.     

Systematics of dusky salamanders, Desmognathus (Caudata: Plethodontidae), in the mountain and Piedmont regions of Virginia and North Carolina, USA

We analysed mitochondrial (cytochrome  b ) nucleotide sequences, nuclear allozyme markers, and morphometric characters to investigate species boundaries and phylogenetic relationships among dusky salamanders ( Desmognathus ) in the southern Blue Ridge and adjacent Piedmont Physiographic Provinces of Virginia and North Carolina. Our results revealed four distinct mitochondrial DNA clades that are also characterized by distinct allozyme markers. One clade consists of sequences derived from populations distributed from New England to south-western Virginia that are referable to Desmognathus fuscus Rafinesque, 1820, although there is considerable sequence and allozyme divergence within this clade. A second clade consists of sequences derived from populations referable to Desmognathus planiceps Newman, 1955, a form that we resurrect from its long synonymy under D. fuscus . Desmognathus planiceps and D. fuscus also differ in mandibular tooth shape. Two other cytochrome  b sequences recovered from populations along the Blue Ridge escarpment in southern Virginia are quite divergent from those of the previous two clades, and these populations may represent yet another distinct species. Sequences from a population in the Brushy Mountains in the Piedmont of northern North Carolina are similar to those of Desmognathus carolinensis . Population groupings indicated by allozyme data generally correspond to the cytochrome  b clades. Cryptic diversity in Appalachian desmognathan salamanders clearly requires further study. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society , 2008, 152 , 115–130.     

High levels of mitochondrial cytochrome b divergence in annual killifishes of the genus Cynolebias (Cyprinodontiformes, Rivulidae)

Phylogenetic relationships based on 324 base pairs of the mitochondrial cytochrome b gene were examined in 14 species of the genus Cynolebias. The monophyly of the genus relative to three outgroup taxa belonging to the family Rivulidae was supported by the sequence data. Bootstrap values corroborated the existence of intrageneric monophyletic units in both parsimony and Neighbour-joining analyses. These include a Cynolebias bellottii-C. wolterstorffl clade, a C. adloJfi-2, C. duraznensis, C. viarius and C adloffi-1 group, and a C. gymnoventris, C. luteoflammulatus pair and the strongly supported assemblage that includes C. prognalhus and C. cheradophilus clade. Phylogenetic relationships remain poorly supported for C. nigripinnis, C. qffinis and C. alexandri, and unresolved among the previous ingroup clades. Cytochrome b sequences reveal an unexpectedly high level of divergence among species of the genus Cynolebias. Consequently, cytochrome b shows good resolution of recent cladogenetic events but limited phylogenetic information at deeper nodes. High levels of sequence divergence span a broad range within Cynolebias. The highest sequence divergence (c. 28%) occurred among C. antenori and the remaining species of the genus. The minimum divergence value (4.5%) is exhibited by sympatric species C. cheradophilus and C. prognathus.     

Karyotype differentiation in Chromaphyosemion killifishes (Cyprinodontiformes, Nothobranchiidae). II: cytogenetic and mitochondrial DNA analyses demonstrate karyotype differentiation and its evolutionary direction in C. riggenbachi

African killifishes of the genus Chromaphyosemion show a high degree of phenotypic and karyotypic diversity. The latter is especially pronounced in C. riggenbachi, a morphologically defined species restricted to a small distribution area in Cameroon. This study presents a detailed reconstruction of karyotype differentiation within C. riggenbachi using conventional Giemsa staining and sequential chromosome banding as well as a phylogenetic analysis based on part of the mitochondrial (mt) cytochrome b gene from eleven populations. The cytogenetic analysis revealed differences in chromosome morphology, banding patterns and/or diploid chromosome number (2n) among all populations examined. Diploid number ranged from 2n = 20 to 2n = 36 and varied mainly among populations, while C-banding patterns and NOR phenotypes showed fixed differences among populations as well as some variability within populations. The mtDNA analysis disclosed five clearly differentiated haplotype groups. Mapping the karyotype data onto the mtDNA dendrogram revealed a decrease in 2n from the most basal to the most derived groups, thus demonstrating a reduction of 2n during their evolutionary history. Our results indicate that karyotype differentiation involved Robertsonian fusions as well as non-Robertsonian processes. Causes of the high karyotypic variability may include an elevated chromosomal mutation rate as well as certain features of the ecology and mating system that could facilitate the fixation of chromosomal rearrangements. The pattern of karyotype and haplotype differentiation and the results of previous crossing experiments suggest incipient speciation in C. riggenbachi.     

Mitochondrial DNA sequence variation and phylogeography among Salmo trutta L. (Greek brown trout) populations

To investigate the phylogenetic relationships and geographical structure among brown trout S. trutta L. populations from the South Adriatic-Ionian and Aegean sea basins, mitochondrial DNA nucleotide sequence comparisons were used. A 310-base-pair (bp) segment of the control region (D-loop), and an additional 280-bp segment of the cytochrome b gene were sequenced from representatives of 13 brown trout populations. Phylogenetic analyses, conducted after combining the data presented with published data from other Eurasian brown trout, revealed four major phylogenetic groups, three of which were found widely distributed within the southern Balkan region. The phylogeographical patterns revealed by mtDNA represent one of the few cases where phylogenetic discontinuity in a gene tree exists without obvious geographical localization within a species' range and has most likely resulted from the differentiation of the major mtDNA clades during Messinian or early Pleistocene times. Finally, the genetic relationships among the populations suggested by mtDNA were generally not in accordance with either allozyme or morphological data.     

Genetic characterization and phylogeography of the wild boar Sus scrofa introduced into Uruguay

The European wild boar Sus scrofa was first introduced into Uruguay, in southern South America during the early decades of the last century. Subsequently, and starting from founder populations, its range spread throughout the country and into the neighbouring Brazilian state Rio Grande do Sul. Due to the subsequent negative impact, it was officially declared a national pest. The main aim in the present study was to provide a more comprehensive scenario of wild boar differentiation in Uruguay, by using mtDNA markers to access the genetic characterization of populations at present undergoing rapid expansion. A high level of haplotype diversity, intermediate levels of nucleotide diversity and considerable population differentiation, were detected among sampled localities throughout major watercourses and catchment dams countrywide. Phylogenetic analysis revealed the existence of two different phylogroups, thereby reflecting two deliberate introduction events forming distantly genetic lineages in local wild boar populations. Our analysis lends support to the hypothesis that the invasive potential of populations emerge from introgressive hybridization with domestic pigs. On taking into account the appreciable differentiation and reduced migration between locales in wild boar populations, management strategies could be effective if each population were to be considered as a single management unit.     

Phylogeography of the Calonectris shearwaters using molecular and morphometric data

We investigated phylogenetic relationships and the biogeographic history of the Calonectris species complex, using both molecular and biometric data from one population of the Cape Verde shearwater Calonectris edwardsii (Cape Verde Islands), one from the streaked shearwater C. leucomelas (western Pacific Ocean) and 26 from Cory's shearwater populations distributed across the Atlantic (C. d. borealis) and the Mediterranean (C. d. diomedea). The streaked shearwater appeared as the most basal and distant clades, whereas the genetic divergences among the three main clades within the Palearctic were similar. Clock calibrations match the first speciation event within Calonectris to the Panama Isthmus formation, suggesting a vicariant scenario for the divergence of the Pacific and the Palearctic clades. The separation between the Atlantic and Mediterranean clades would have occurred in allopatry by range contraction followed by local adaptation during the major biogeographic events of the Pleistocene. The endemic form from Cape Verde probably evolved as a result of ecological divergence from the Mediterranean subspecies. Finally, one Mediterranean population (Almeria) was unexpectedly grouped into the Atlantic subspecies clade, both by genetic and by morphometric analyses, pointing out the Almeria-Oran oceanographic front (AOOF) as the actual divide between the two Cory's shearwater subspecies. Our results highlight the importance of oceanographic boundaries as potentially effective barriers shaping population and species phylogeographical structure in pelagic seabirds.     

Phylogenetic divergence in leatherside chub (Gila copei) inferred from mitochondrial cytochrome b sequences

We examined intra-specific phylogenetic relationships in leatherside chub, Gila copei. The complete mitochondrial (mt) cytochrome b gene (1140 bp) was sequenced for 30 individuals from 10 populations that span the geographical distribution of this species. Traditional phylogenetic analyses revealed two deeply divergent and evolutionarily distinct mtDNA clades that are geographically separated in northern and southern drainage basins. Interpopulation sequence variation between clades ranged from 7.7 to 8.1%. The northern clade was genetically more similar and phylogenetically more closely related to the selected out-group Lepidomeda m. mollispinus than to the southern clade, suggesting that the taxonomy of this species may require revision. Sequence variation among populations within clades ranged from 0 to 0.3% in the north and from 0 to 0.7% in the south. Statistical parsimony was used to construct phylogenetic networks of haplotypes within clades. Nested clade analysis revealed that geographical fragmentation has played an important role in genetic structuring within northern and southern clades.     

Phylogeny, genetic variability and colour polymorphism of an emerging animal model: the short-lived annual Nothobranchius fishes from southern Mozambique

Nothobranchius are a group of small, extremely short-lived killifishes living in temporary savannah pools in Eastern Africa and that survive annual desiccation of their habitat as dormant eggs encased in dry mud. One mitochondrial (COI) and three nuclear (CX32.2, GHITM, PNP) loci were used to investigate the phylogenetic relationship of Nothobranchius species from southern and central Mozambique. This group shows marked variation in captive lifespan at both the inter- and intraspecific levels; lifespan varies from a few months to over a year. As their distribution encompasses a steep gradient between semi-arid and humid habitats, resulting in contrasting selection pressures on evolution of lifespan and associated life history traits, Mozambican Nothobranchius spp. have recently become a model group in studies of ageing, age-related disorders and life history evolution. Consequently, intraspecific genetic variation and male colour morph distribution was also examined in the recovered clades. Using Bayesian species tree reconstruction and single loci analyses, three large clades were apparent and their phylogenetic substructure was revealed at the inter- and intra-specific levels within those clades. The Nothobranchius furzeri and Nothobranchius orthonotus clades were strongly geographically structured. Further, it was demonstrated that male colour has no phylogenetic signal in N. furzeri, where colour morphs are sympatric, but is associated with two reciprocally monophyletic groups in Nothobranchius rachovii clade, where colour morphs are parapatric. Finally, our analysis showed that a polymorphism in the Melanocortin1 receptor gene (which controls pigmentation in many vertebrates and was a candidate gene of male colouration in N. furzeri) is unrelated to colour phenotypes of the study species. Our results raise significant implications for future comparative studies of the species and populations analysed in the present work.     

Salenthydrobia gen. nov. (Rissooidea: Hydrobiidae): a potential relict of the Messinian salinity crisis

The Messinian salinity crisis (MSC) occurred synchronously throughout the Mediterranean basin about 5.96 ± 0.02 Mya and represents one of the most dramatic oceanic changes since the early Miocene. It is thought that the concomitant environmental changes brought about isolation of faunas and the development of endemism. As part of the search for possible speciation events triggered by the MSC, the author studied 38 populations of hydrobiine snails from the Mediterranean and Black Sea, including three populations from the Salentina Peninsula, Italy. Partial sequences (COI, 16S) and anatomical data were used to test the taxonomic and phylogenetic status of the peninsular populations. A maximum likelihood analysis of 11 hydrobiine taxa revealed five clades and lineages, four of which corresponded to previously recognized genera: Adriohydrobia , Hydrobia , Peringia , Ventrosia . The fifth clade was formed by haplotypes of the peninsular populations, which are characterized by distinct male and female reproductive systems. Based on molecular and anatomical data, these populations are considered to represent a new species, Salenthydrobia ferrerii , belonging to a new genus, Salenthydrobia . Ecological and biogeographical data for S. ferrerii strongly support a correlation between its origin and the MSC. Based on an island age of 5.33 Myr and a population divergence of 0.0973 ± 0.0114, the COI molecular clock rate for the Salenthydrobia and Peringia clades would be 1.83 ± 0.21% population divergence per Myr. The genetic diversity of S. ferrerii , its phylogenetic relationships, and the validity of the proposed local molecular clock rate are discussed.     

Patterns of genetic differentiation in the <Emphasis Type="Italic">Gymnogeophagus gymnogenys</Emphasis> species complex,a neotropical cichlid from South American basins

The neotropical cichlid genus Gymnogeophagus is distributed in the Río de la Plata basin and in Dos Patos and Merín coastal lagoons on the border between Uruguay and southern Brazil. A phylogeographic approach based on mitochondrial cytochrome b analysis was performed to assess the patterns and processes of differentiation in this taxon. Gymnogeophagus gymnogenys showed high haplotype diversity (H = 0.992) and corrected mtDNA genetic distances ranged from 0 to 5.3%. Our analyses yielded robust support for the existence of four monophyletic groups within G. gymnogenys from the analyzed basins. No correlation between the aforementioned clades and geographic structure was found, since individuals belonging to different phylogenetic clades inhabit the same locality. The phylogeographic approach presented here showed that these four phylogroups (1, 2, 3 and 4) were sister groups. Our present findings would corroborate that G. gymnogenys could be integrated by different phylogenetic lineages, showing an explosive differentiation pattern and confirming the hypothesis that this taxon constitutes a species complex.     

Early development in the annual fish Cynolebias viarius

The detailed characteristics and timing of early development in Cynolebias viarius , an annual fish endemic to Uruguay, were obtained from in vivo observations of embryos cultured under standardized laboratory conditions. The chorion ultrastructure is reported for this species for the first time. Comparison with previous data from other annual fishes showed that C. viarius exhibited the longest early development including cleavage, blastula formation and epiboly.