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The Trichomycterinae is a diversified catfish clade exhibiting numerous ecological specializations, thus offering fine opportunities for studies on vertebrate evolution, but insufficient knowledge on phylogenetic relationships and taxonomy impedes advanced evolutionary studies. Recent studies have contributed to a better taxonomical resolution of some trichomycterine groups, but several taxa remain poorly known, with taxonomical placement still undetermined. This study is directed to two similar undescribed species with unclear generic placement, collected in the savannahs of central Brazil, in the headwaters of the two most important South American river systems, the Amazon and Parana basins central Brazil. A multigene phylogeny integrated to a comparative osteological analysis supports a clade, with the new species forming a subclade sister to a trichomycterine subclade endemic to southern South America. The comparative osteological analysis provided some characters from the jaw suspensorium, opercular series, branchial arches, neurocranium, and pectoral girdle that are useful to diagnose members of the clade. The two species may be distinguished by meristic data, morphometrics, osteology, and colour pattern. Following recent efforts for a better understanding about diversity and relationships of trichomycterines, the present study provides phylogenetic and morphological basis for the taxonomical resolution of this important trichomycterine clade. However, much more research is necessary to improve trichomycterine taxonomy, including further studies providing a proper generic classification encompassing the whole Trichomycterinae.

The origin of melanism has been often associated with cryptic habits, driven by selection for camouflage, sometimes independently arising in closely related species. Field studies in Atlantic Forest rivers of south-eastern Brazil have indicated the occurrence of three melanic species in the catfish genus Trichomycterus: Trichomycterus immaculatus, Trichomycterus nigricans, and T. quintus Costa sp. nov. We performed a molecular analysis using two nuclear and two mitochondrial genes (2811 bp) for 62 trichomycterines and eight outgroups, in which monophyly of a group here named as the T. nigricans-nigroauratus clade is highly supported. It comprises two subclades, the T. nigricans group (containing Trichomycterus caipora, T. immaculatus, T. nigricans, and Trichomycterus santaeritae) and the Trichomycterus nigroauratus group (containing Trichomycterus maculosus, T. nigroauratus, and T. quintus). Ancestral state reconstructions indicate that melanism has arisen independently in each species exhibiting overall black colouration, which may be favoured by the strict nocturnal habits of these three species. However, whereas in the T. nigricans group cryptic habits precede the origin of melanism, in the T. nigroauratus group these events occur simultaneously during the evolution of T. quintus. Two melanic species, T. immaculatus and T. quintus, are sympatric but inhabit different habitats, suggesting some ecological divergence. Oppositely to the crypsis evolutionary trend, the analysis indicates a reversal to diurnal habits in the psammophilic T. santaeritae. This species, uniquely among congeners of the T. nigricans-nigroauratus clade, has a colour pattern consisting of dark brown blotches on a pale yellow colour ground and relatively large eyes, allowing sandy habitat camouflage and visual orientation during foraging activity, respectively, interpreted as adaptive morphological changes parallelly found in other diurnal psammophilic trichomycterids.

This study is primarily directed to the most poorly known species of the genus Trichomycterus, comprising five nominal species (T. florensis, T. immaculatus, T. nigricans, T. paquequerensis and T. ...

Trichomycterid catfishes of the Sarcoglanidinae, which includes small-sized species living in sandy river substratum, have been poorly represented in collections. We herein describe two new...

The Trichomycterinae is among the most diverse catfish groups in the world, with interesting ecological specializations; however, the taxonomy of Trichomycterinae is still problematic, mainly due to unclear limits and diagnoses of the eight included genera. Herein we infer the phylogenetic position and generic placement of Trichomycterus payaya, a recently described species from north-eastern Brazil, with unique morphological features among congeners. A multigene analysis including fragments of two nuclear and two mitochondrial genes (total of 2974 bp) for 53 trichomycterine taxa and three outgroups clearly supports inclusion of T. payaya in Ituglanis, a trichomycterine genus containing 29 valid species, which is corroborated by osteological characters. An examination of osteological characters used to diagnose Ituglanis in its original description indicates that a rudimentary or absent posterior cranial fontanel is synapomorphic for Ituglanis, but this is reversed in I. payaya; an anteriorly directed antero-lateral extremity of the sphenotic-prootic-pterosphenoid complex and a narrow and long lateral process of the parurohyal are synapomorphic for Ituglanis, but homoplastically occurring in other trichomycterids; and the presence of a deep medial concavity on the autopalatine is ambiguous to diagnose Ituglanis by occurring in other closely related trichomycterine taxa. Ituglanis is also distinguishable from other trichomycterines by a unique shape of the metapterygoid and by a reduced number of ribs. The analysis supports a clade, comprising I. payaya and I. paraguassuensis, that is endemic to the Chapada Diamantina, in the semiarid Caatinga of north-eastern Brazil, reinforcing the importance of this region for trichomycterid diversification. By integrating molecular analysis and comparative morphology, the present study provides a more solid basis for delimiting Ituglanis, creating a better taxonomical resolution of the Trichomycterinae, although much more research is necessary to reach a consistent generic classification for the entire subfamily.

A recent collection of the seasonal killifish found Leptopanchax splendens c. 5 km from the type locality, 74 years after its last record. The species was historically common in its type locality, the Estrela River basin in south-eastern Brazil, until 1950, after which it was not encountered and thought to have become extinct due to widespread deforestation and urbanization in the region. Despite the rediscovery, this study finds that other recently published reports of L. splendens are misidentifications.

Three distinct and independent molecular-based species delimitation analyses were performed among the species and populations included within theAustraloherosautranigroup, based on sequences of the mitochondrial gene Cytochrome b: a tree-based method proposed by Wiens and Penkrot (WP), a Character-based DNA Barcoding (CBB) and coalescent species delimitation method termed the Bayesian Implementation of the Poisson tree processes (bPTP). The congruence of WP and CBB delimited 11 independent lineages (species), while the bPTP delimited just nine lineages. We did not favour any of the methods, and we considered the possibility of two slightly variant scenarios. A time-calibrated phylogenetic analysis is proposed based on the predominant congruence of the results of these three species delimitation methods herein applied. The monophyly of theA.autranispecies group was highly supported with maximum node support value and diagnosed by 11 nucleotide substitutions. The sister clade of theA.autranispecies group is the clade comprisingA.sp. Timbé do Sul andA.minuano. The phylogenetic analysis supports three main clades within theA.autranispecies group, supported by maximum node support value, with the Southern Mata Atlântica clade as the most basal clade. Divergence time estimates indicate that the diversification of theAustraloherosoriginated during the early Neogene, but only in the late Neogene did the processes of diversification in the southeast and north regions occur. Diversification within theAustraloherosautranispecies group occurred synchronically for the three main clades during the beginning of the Quaternary. It is demonstrated that molecular characters are valuable tools for species recognition, particularly in speciose groups with inconspicuous or difficult to record morphological characters. The resulting phylogeny of theAustraloherosautranigroup is highly compatible with the geological and biogeographic scenarios proposed for the Neogene and Quarternary shaping of the extant river basins of eastern Brazil. Despite the origin of theA.autranigroup being dated to the late Miocene, species level diversification occurred in the Pleistocene and was probably driven by headwater capture events and sea-level fluctuations.

A high concentration of endemic species of seasonal killifishes has been recorded for a small area encompassing the highland plateaus associated with the upper section of the Carnaúba de Dentro River drainage and adjacent drainages of the middle section of the São Francisco River basin, northeastern Brazil. The present study is primarily directed to the taxonomy of theH.fulminantisspecies complex in this region, and describes habitat decline and extirpation of natural killifish populations recorded in field studies between 1993 and 2017. Both morphological characters and molecular species delimitation methods using single-locus models (GMYC and bPTP) support recognition of two closely related endemic species,H.fulminantisandH.splendissimusCosta,sp. n.The new species is distinguished from other congeners of theH.fulminantiscomplex by having a red pectoral fin in males, well-developed filamentous rays on the tips of the dorsal and anal fins in adult males, and the second proximal radial of the dorsal fin between the neural spines of the 8thand 9thvertebrae in males. Most recent field inventories indicated possible local extinction of populations ofH.fulminantisandH.splendissimusin the studied area, but additional field studies should be made in other parts of the upper Carnaíba de Dentro River basin to evaluate the current conservation status of these species.

A great diversity of animal species adapted to life in the semi-arid Caatinga of northeastern Brazil, including seasonal killifishes, has been reported in the last three decades. More recently, field and molecular data have shown a high occurrence of cryptic species. The killifish group herein analysed, the Hypsolebiasmagnificus species complex, is endemic to the middle and southern portion of the Caatinga, occupying about 120 km along the floodplains of the middle São Francisco River and some adjacent tributaries. Species of this complex are rare and presently considered threatened with extinction, being uniquely found in pools protected by trees and bushes. Single-locus delimitation methods were used to test species limits of populations displaying different colour patterns along the whole distribution of the complex. All analyses consistently supported the three nominal species and two new, herein described: H.gardneri Costa, sp. n., from the floodplains of the middle São Francisco River and H.hamadryades Costa, sp. n., from the Gorotuba River floodplains. The phylogenetic analysis highly supports H.hamadryades as sister to a clade comprising H.gardneri and H.harmonicus. Our field observations suggest that H.hamadryades is a miniature species. This study indicates that the H.magnificus complex comprises cryptic species apparently endemic to small areas and extremely vulnerable to environmental changes, deserving high concern.

The Caatinga is the largest nucleus of seasonally dry tropical forests in South America, but little is known about the evolutionary history and biogeography of endemic organisms. Evolutionary diversification and distribution of terrestrial vertebrates endemic to the Caatinga have been explained by palaeogeographical Neogene episodes, mostly related to changes in the course of the Sao Francisco River, the largest river in the region. Our objective is to estimate the timing of divergence of two endemic groups of short-lived seasonal killifishes inhabiting all ecoregions of the Caatinga, testing the occurrence of synchronic events of spatial diversification in light of available data on regional palaeogeography. We performed independent time-calibrated phylogenetic molecular analyses for two clades of sympatric and geographically widespread seasonal killifishes endemic to the Caatinga, the Hypsolebias antenori group and the Cynolebias alpha-clade. Our results consistently indicate that species diversification took place synchronically in both groups, as well as it is contemporary to diversification of other organisms adapted to life in the semi-arid Caatinga, including lizards and small mammals. Both groups originated during the Miocene, but species diversification started between the Late Miocene and Early Pliocene, when global cooling probably favoured the expansion of semi-arid areas. Synchronic diversification patterns found are chronologically related to Tertiary palaeogeographical reorganizations associated to continental drift and to Quaternary climatic changes, corroborating the recent proposal that South American biodiversity has been continuously shaped between the Late Paleogene and Pleistocene.

Morphological characters and phylogenetic trees generated by analyses of segments of two mitochondrial genes cytochrome b and cytochrome c oxidase I support recognition of three new species of the ‘Geophagus’ brasiliensis species group from coastal basins of northeast Brazil. All new species were diagnosed by exclusive morphological characters and exclusive nucleotide transformations. Geophagusrufomarginatus sp. n., from the Rio Buranhém Basin, is distinguished from all other species of the group by dorsal-fin lappets with red edges, the presence of longitudinal series of small light blue spots between the anal-fin spines and rays, and non-denticulated gill-rakers; it is closely related to G.brasiliensis and G.iporangensis. Geophagusmultiocellus sp. n., from the Rio de Contas Basin, is distinguished from all other species of the group by having small pale blue spots with minute bright blue dots at their centres, that are often vertically coalesced to form short bars on the caudal fin. Geophagussantosi sp. n., from the Rio Mariana Basin, is distinguished from all other species of the group by having blue stripes parallel to the dorsal and anal fin rays on their longest portions. Geophagusmultiocellus and G.santosi belong to the same clade of G.itapicuruensis. The clade composed by the Rio Paraguaçu Basin species was recovered as the sister group of the other species of the ‘G.’ brasiliensis species group.

Trichomycterus comprises about 170 valid species, but its monophyly has been challenged in the last decades. Bayesian Inference and Maximum Likelihood analyses comprehending mitochondrial genes COI and CYTB and nuclear genes GLYT, MYH6 and RAG2 from 71 Trichomycterinae terminal taxa and eight outgroups were performed. The analyses highly supports a clade containing Trichomycterusnigricans, the type species of the genus, and several other congeners endemic to eastern and northeastern Brazil, herein considered as the genus Trichomycterus, the sister clade the southern Brazil and adjacent areas clade; the latter clade comprises two subclades, one comprising species of the genus Scleronema and another comprising species previously placed in Trichomycterus, herein described as a new genus. Cambevagen. n. is distinguished from all other trichomycterines by the presence of a bony flap on the channel of the maxillo-dentary ligament, the interopercle shorter than the opercle, a deep constriction on the basal portion of the antero-dorsal arm of the quadrate, absence of teeth in the coronoid process of the dentary, the maxilla shorter than the premaxilla, the cranial fontanel extending from the the medial posterior of frontal to the medial region of supraoccipital, and absence of the postorbital process of the sphenotic-prootic-pterosphenoid.

Rondonacara, gen. nov. is herein described as a member of the tribe Cichlasomatini, being considered as the sister taxon of Laetacara. The new genus differs from all the other genera of Cichlasomatini by having transversal streaks originated at the insertion of spines and softs rays of dorsal fin. Additionally, the new genus differs from the other genera of the Cichlasomatini by its unique combination of character states: longitudinal stripe of the flank horizontally oriented, ending directed to caudal-fin base; conspicuous suborbital bar; caudal-fin base spot on the middle of the fin and not ocellated; preopercle without scales; two supra-neural; and uniserial predorsal squamation. Rondonacara is monotypic, including only Rondonacara hoehnei, re-described herein. This species was originally described from a tributary of the Araguaia River, Mato Grosso state, with unprecise locality. Currently, Rondonacara hoehnei is known only from a single locality, a tributary of the das Mortes River, Araguaia River basin, in the Brazilian Cerrado. The restrict distribution area, and the widespread and accelerated destruction of natural habitat strongly indicate that Rondonacara hoehnei is critically endangered

The rich biological diversity of South America has motivated a series of studies associating evolution of endemic taxa with the dramatic geologic and climatic changes that occurred during the Cainozoic. The organism here studied is the killifish tribe Cynolebiini, a group of seasonal fishes uniquely inhabiting temporary pools formed during the rainy seasons. The Cynolebiini are found in open vegetation areas inserted in the main tropical and subtropical South American phytogeographical regions east of the Andes. Here, we present the first molecular phylogeny sampling all the eight genera of the Cynolebiini, using fragments of two mitochondrial and four nuclear genes for 35 species of Cynolebiini plus 19 species as outgroups. The dataset, 4448bp, was analysed under Bayesian and maximum likelihood approaches, providing a relatively well solved tree, which retrieves high support values for the Cynolebiini and most included clades. The resulting tree was used to estimate the time of divergence in included lineages using two cyprinodontiform fossils to calibrate the tree. We further investigated historical biogeography through the likelihood-based DEC model. Our estimates indicate that divergence between the clades comprising New World and Old World aplocheiloids occurred during the Eocene, about 50Mya, much more recent than the Gondwanan fragmentation scenario assumed in previous studies. This estimation is nearly synchronous to estimated splits involving other South American and African vertebrate clades, which have been explained by transoceanic dispersal through an ancient Atlantic island chain during the Palaeogene. We estimate that Cynolebiini split from its sister group Cynopoecilini in the Oligocene, about 25Mya and that Cynolebiini started to diversify giving origin to the present genera during the Miocene, about 20-14Mya. The Cynolebiini had an ancestral origin in the Atlantic Forest and probably were not present in the open vegetation formations of central and northeastern South America until the Middle Miocene, when expansion of dry open vegetation was favoured by cool temperatures and strike seasonality. Initial splitting between the genera Cynolebias and Simpsonichthys during the Miocene (about 14Mya) is attributed to the uplift of the Central Brazilian Plateau.

Internal fertilization is a widespread mode of reproduction in chondrichthyans and tetrapods, but uncommon in actinopterygian fishes. In killifishes of the suborder Aplocheiloidei, internal fertilization is restricted to two genera, Campellolebias and Cynopoecilus, both containing species adapted to life in seasonal pools of subtropical South America and exhibiting elaborated inseminating structures. Phylogenetic studies involving these genera are scarce and limited to morphological characters and fragments of mitochondrial DNA sequences available for a few taxa, providing incongruent results and thus impeding hypotheses on the evolution of insemination and related morphological traits. We analyzed three nuclear loci (GLYT1, ENC1, Rho) for 13 aplocheiloid taxa obtaining the first well-supported phylogeny for cynopoecilines, thus providing a significant background to interpret evolutionary changes within the group. Like in killifishes of the suborder Cyprinodontoidei, the evolution of internal fertilization in aplocheiloids is associated with deep changes in the structure of male anal fin. The phylogenetic analyses indicate that internal fertilization corresponds to a single evolutionary event during the evolution of aplocheiloid killifishes. The analyses also indicate that male specialized muscle characters, comprising a muscular ejaculatory pump in the urogenital region and hypertrophied inclinatores and depressores anales, arose in the ancestor of the clade comprising Campellolebias and Cynopoecilus. On the other hand, anal fin specialized structures including the male inseminating tube of Campellolebias and the male inseminating fan of Cynopoecilus evolved independently in each genus.

The Cynopoecilus melanotaenia complex is a morphologically homogeneous killifish group, endemic from an area encompassing southern Brazil and northeastern Uruguay. It presently comprises four valid species: C. melanotaenia, the type species of the genus, and C. fulgens, C. intimus, and C. nigrovittatus. Cynopoecilus feltrini, n. sp., from the lower Tubarão river basin, southern Brazil, is distinguished from all congeners of the C. melanotaenia complex by having frontal E-scales medially overlapped, branchiostegal region orangish red in males and dorsum with few dark brown spots above opercular region. A phylogenetic tree derived from the analysis of a fragment of the mitochondrial gene cytochrome c oxidase subunit I (681 bp) indicates that C. feltrini is a member of the clade that includes all species of the C. melanotaenia complex except C. melanotaenia, as well as that C. feltrini is the sister group of a clade comprising C. fulgens and C. nigrovittatus.