Chromosomal analysis of two Acanthodoras species (Doradidae, Siluriformes): Insights into the oldest thorny catfish clade and its karyotype evolution.

The Doradidae fishes constitute one of the most diverse groups of Neotropical freshwater environments. Acanthodoradinae is the oldest lineage and the sister group to all other thorny catfishes, and it includes only the genus Acanthodoras. The diversity of Acanthodoras remains underestimated, and the use of complementary approaches, including genetic studies, is an important step to better characterize this diversity and the relationships among the species within the genus. Therefore, we conducted a comprehensive analysis using conventional cytogenetic techniques and physical mapping of three multigene families (18S and 5S ribosomal DNA [rDNA], U2 small nuclear DNA [snDNA]) and four microsatellite motifs, namely (AC)n, (AT)n, (GA)n, and (GATA)n, in two sympatric species from the Negro River: Acanthodoras cataphractus and Acanthodoras cf. polygrammus. We found significant differences in constitutive heterochromatin (CH) content, distribution of the microsatellite (AT)n, and the number of 5S rDNA and U2 snDNA sites. These differences may result from chromosome rearrangements and repetitive DNA dispersal mechanisms. Furthermore, the characterization of the diploid number (2n) of these Acanthodoras species enables us to propose 2n = 58 chromosomes as the plesiomorphic 2n state in Doradidae based on ancestral state reconstruction. Acanthodoradinae is the oldest lineage of the thorny catfishes, and knowledge about its cytogenetic patterns is crucial for disentangling the karyotype evolution of the whole group. Thus, this study contributes to the understanding of the mechanisms behind chromosome diversification of Doradidae and highlights the importance of Acanthodoradinae in the evolutionary history of thorny catfishes.

Exploring hidden diversity: Molecular insights into the Leporinus species of the rivers of the Brazilian states of Maranhão and Piauí.

The present study delved into the world of hidden diversity by examining specimens identified as Leporinus piau from the river basins of the northern Brazilian states of Maranhão and Piauí. Using genetic analyses that combined data from three mitochondrial markers and one nuclear marker, the study identified two well-supported groups, reinforcing the findings of previous publications. The first group, found in samples from the Itapecuru, Mearim, Turiaçu, and Pericumã basins, in Maranhão, appears to represent a relatively ancient diversification and the possibility of concealed cryptic diversity. The second group, comprising specimens from the Parnaíba (Piauí) and Mearim (Maranhão) basins, appears to have resulted from a more recent process of diversification and has a close relationship with Leporinus friderici from the type locality. Our findings not only confirm the existence of a complex scenario of cryptic diversity in the genus Leporinus from the study basins but also underscore the taxonomic inconsistencies within this group of fish. This study offers a comprehensive analysis of the species diversity of the Maranhão and Piauí basins, which are critical regions for the conservation of Amazonian fish, providing valuable insights for the sustainable management and conservation of these fish.

The DNA barcode reveals cryptic diversity and a new record for the genus Leporinus (Characiformes, Anostomidae) in the hydrographic basins of central northern Brazil.

Leporinus is one of the most speciose genera of the order Characiformes, with 81 valid species distributed throughout much of Central and South America. The considerable diversity of this genus has generated extensive debate on its classification and internal arrangement. In the present study, we investigated the species diversity of the genus Leporinus in central northern Brazil, and conclude that six valid species-Leporinus maculatus, Leporinus unitaeniatus, Leporinus affinis, Leporinus venerei, Leporinus cf. friderici, and Leporinus piau-are found in the hydrographic basins of the Brazilian states of Maranhão, Piauí, and Tocantins. We analyzed 182 sequences of the Cytochrome Oxidase subunit I gene, of which, 157 were obtained from Leporinus specimens collected from the basins of the Itapecuru, Mearim, Turiaçu, Pericumã, Periá, Preguiças, Parnaíba, and Tocantins rivers. The species delimitation analyses, based on the ABGD, ASAP, mPTP, bPTP, and GMYC methods, revealed the presence of four distinct molecular operational taxonomic units (MOTUs), identified as L. maculatus, L. unitaeniatus, L. affinis, and L. piau (from the Parnaíba River). The bPTP method restricted L. venerei to a single MOTU, and confirmed the occurrence of this species in the rivers of Maranhão for the first time. The separation of L. cf. friderici into two clades and the subsequent formation of different operational taxonomic units was consistent with polyphyly in this species, which indicates the existence of cryptic diversity. The arrangement of L. cf. friderici and L. piau in two different clades supports the conclusion that the L. piau specimens from Maranhão were misidentified, based on their morphological traits, reflecting the taxonomic inconsistencies that exist among morphologically similar species. Overall, then, the species delimitation methods employed in the present study indicated the presence of six MOTUs-L. maculatus, L. unitaenitus, L. affinis, L. cf. friderici, L. venerei, and L. piau. In the case of two other MOTUs identified in the present study, one (L. venerei) is a new record for the state of Maranhão, and we believe that the other represents a population of L. piau from the basin of the Parnaíba River.

Reconstruction of the Doradinae (Siluriformes-Doradidae) ancestral diploid number and NOR pattern reveals new insights about the karyotypic diversification of the Neotropical thorny catfishes.

Doradinae (Siluriformes: Doradidae) is the most species-rich subfamily among thorny catfishes, encompassing over 77 valid species, found mainly in Amazon and Platina hydrographic basins. Here, we analyzed seven Doradinae species using combined methods (e.g., cytogenetic tools and Mesquite ancestral reconstruction software) in order to scrutinize the processes that mediated the karyotype diversification in this subfamily. Our ancestral reconstruction recovered that 2n=58 chromosomes and simple nucleolar organizer regions (NOR) are ancestral features only for Wertheimerinae and the most clades of Doradinae. Some exceptions were found in Trachydoras paraguayensis (2n=56), Trachydoras steindachneri (2n=60), Ossancora punctata (2n=66) and Platydoras hancockii whose karyotypes showed a multiple NOR system. The large thorny catfishes, such as Pterodoras granulosus, Oxydoras niger and Centrodoras brachiatus share several karyotype features, with subtle variations only regarding their heterochromatin distribution. On the other hand, a remarkable karyotypic variability has been reported in the fimbriate barbells thorny catfishes. These two contrasting karyoevolution trajectories emerged from a complex interaction between chromosome rearrangements (e.g., inversions and Robertsonian translocations) and mechanisms of heterochromatin dispersion. Moreover, we believe that biological features, such as microhabitats preferences, populational size, low vagility and migratory behavior played a key role during the origin and maintenance of chromosome diversity in Doradinae subfamily.

Taxonomic revision of the genus Anostomoides (Characiformes: Anostomidae).

A taxonomic revision of Anostomoides is herein presented based on a comprehensive revision of specimens deposited in fish collections and the literature. The present revision proposes that A. laticeps is a junior synonym of A. atrianalis, while A. passionis is a junior synonym of Leporinus nattereri, and the latter is thus transferred to Anostomoides, forming the new combination Anostomoides nattereri. Principal Components Analysis (PCA) showed that examined specimens form two distinct morphotypes, corroborating the validity of both A. atrianalis and A. nattereri. Anostomoides nattereri is distinguished from A. atrianalis by having four branchiostegal rays (vs. three), three pores in infraorbital one (vs. four), 37-39 pored lateral-line scales (vs. 41-44), lower lip with rounded dermal papillae (vs. forming ridges), three dark  rounded midlateral blotches (vs. three or four vertically elongated blotches and/or a faded dark longitudinal stripe), dark lines between scale series on posterior half of body on specimens smaller than 150 mm SL (vs. dark lines absent), a greater body depth (27.6-36.2 % vs. 24.5-38.7% of SL) and smaller interorbital distance (34.4-53.8 % vs. 42.6-67.3 % of HL). Anostomoides nattereri is distributed across several tributaries of the Amazon and Orinoco rivers, whereas A. atrianalis is widespread throughout the Amazon, Orinoco, and Essequibo basins. The genus Anostomoides is currently diagnosed based on a combination of non-exclusive characters: upturned or slightly upturned mouth with four premaxillary teeth including symphyseal tooth bicuspid or with blunt cutting edge, remaining teeth slightly tricuspid (with medial cuspid distinctly larger); four dentary teeth, symphyseal tooth with truncate cutting edge (without cusps); second tooth with a single large cusp, and two lateral teeth with three or slightly more small cusps.

Integrative approach detects natural hybridization of sympatric lambaris species and emergence of infertile hybrids.

Despite its relevance for ecology, evolution and conservation of species, natural hybridization and hybrids biology are still poorly studied in freshwater fish. Here, we tested the hypothesis that sympatric species Astyanax paranae and A. fasciatus are able to interbreed in the natural environment and presented evidence for the first record of hybridization between these species. We analyzed anatomical traits, gametogenesis, reproductive biology, and genetic variations of the COI and S7 genes of both species and putative hybrids. Intermediate morphometric and meristic features were observed in hybrids when compared to A. paranae and A. fasciatus. Overlap in reproductive season was showed for these species, with greater reproductive activity from August to January, but hybrids did not present any sign of gonadal maturation. Oogonia and perinucleolar follicles as well as spermatogonia and primary spermatocytes were found in hybrids, but previtellogenic and vitellogenic follicles, spermatids, and spermatozoa were absent. Moreover, several alterations in gametogenesis were detected, such as interrupted meiosis in both males and females, vacuolated and degenerated germ cells, increased interstitial tissue, and presence of immune cells. Molecular analyses supported the hypothesis of hybridization between A. paranae and A. fasciatus. Overall, our multidisciplinary approach also provides strong evidence that hybrids are infertile.

Integrative taxonomy supports new candidate fish species in a poorly studied neotropical region: the Jequitinhonha River Basin.

Molecular identification through DNA barcoding has been proposed as a way to standardize a global biodiversity identification system using a partial sequence of the mitochondrial COI gene. We applied an integrative approach using DNA barcoding and traditional morphology-based bioassessment to identify fish from a neotropical region possessing low taxonomic knowledge: the Jequitinhonha River Basin (Southeastern Brazil). The Jequitinhonha River Basin (JRB) has a high rate of endemism and is considered an area of high priority for fish conservation, with estimates indicating the presence of around 110 native and non-indigenous species. DNA barcodes were obtained from 260 individuals belonging to 52 species distributed among 35 genera, 21 families and 6 orders, including threatened and rare species such as Rhamdia jequitinhonha and Steindachneridion amblyurum. The mean Kimura two-parameter genetic distances within species, genera and families were: 0.44, 12.16 and 20.58 %, respectively. Mean intraspecific genetic variation ranged from 0 to 11.43 %, and high values (>2 %) were recovered for five species. Species with a deep intraspecific distance, possibly flagging overlooked taxa, were detected within the genus Pimelodella. Fifteen species, only identified to the genus level, had unique BINs, with a nearest neighbor distance over 2 % and therefore, potential new candidate species supported by DNA barcoding. The integrative taxonomy approach using DNA barcoding and traditional taxonomy may be a remedy to taxonomy impediment, accelerating species identification by flagging potential new candidate species and to adequately conserve the megadiverse neotropical ichthyofauna.

External Morphology of Lophiosilurus alexandri Steindachner, 1876 during Early Stages of Development, and Its Implications for the Evolution of Pseudopimelodidae (Siluriformes).

Pseudopimelodidae are Neotropical catfishes characterized by having slightly to strongly depressed body in fully developed specimens. The largest species of the family with 500 mm SL, Lophiosilurus alexandri, experiences impressive changes in body shape during development, becoming extremely depressed when fully developed. Accordingly, Lophiosilurus alexandri is an ideal species to observe the morphological changes during ontogeny, and to seek solid interpretations on the polarity of characters. Specimens of distinct larval periods (yolk sac, flexion and postflexion; n = 186 specimens) and juvenile stages (n = 20) were analyzed. Changes in body shape, position of mouth and eye, morphology of fins and pigmentation were observed during the development of Lophiosilurus. Larvae (5.7-11.2 mm standard length) had pigmentation concentrated on the head and parts of body, eyes small and pigmented, short barbels, and well-developed finfold. Juveniles (15.9-28.1 mm standard length) had body shape similar to adult, with head depressed and bearing bony ridges, large mouth, dorsally-oriented eyes, small barbels and well-developed shoulder bulges (cleithral width). The greatest morphological changes in the development of L. alexandri occurred during the postflexion larval stage. Relative to standard length, measurements of snout length, head depth and body depth are smaller in juveniles than in larvae, but body width is larger. New interpretations on the phylogenetic characters related to these changes are provided in view of the two alternative hypotheses of the evolution of Pseudopimelodidae.

A new species of the catfish genus Centromochlus (Siluriformes: Auchenipteridae: Centromochlinae) from the upper rio Paraná basin, Brazil

Centromochlus comprises twelve species, distributed in the main inland watersheds of South America, including the Orinoco, Essequibo, coastal rivers of Suriname, Amazon, upper Paraná and São Francisco basins. The new species is described from the upper rio Paraná based on material collected in 1965 during the construction of the UHE Ilha Solteira, São Paulo, Brazil. The new species is easily distinguished from all congeners due to absence of adipose fin, a condition otherwise restricted to Gelanoglanis nanonocticolus, among centromochlin catfishes. The new species comprises small catfishes (adults ranging from 35 to 39 mm SL), in which modified anal fin of males is devoid of denticulations or spines, and most posterior rays reduced in length. In addition, Tatia simplex Mees is transferred to Centromochlus and its generic reassignment discussed.(AU)

Centromochlus abriga doze espécies, com registros para os principais sistemas fluviais de águas interiores da América do Sul, como o Orinoco, Essequibo, rios costeiros do Suriname, Amazonas, alto Paraná e São Francisco. A nova espécie é descrita do alto rio Paraná, a partir de material coletado em 1965 durante a construção da UHE Ilha Solteira, São Paulo, Brasil. A espécie nova distingue-se facilmente de todas as suas congêneres pela ausência de nadadeira adiposa, uma condição até então restrita à Gelanoglanis nanonocticolus, dentre os bagres centromochlíneos. A nova espécie inclui pequenos bagres (adultos entre 35 e 39 mm CP), nos quais a nadadeira anal modificada em machos é desprovida de dentículos ou espinhos, e com os raios mais posteriores de tamanho reduzido. Adicionalmente, Tatia simplex Mees é transferida para Centromochlus e é fornecida uma discussão acerca de seu posicionamento genérico.(AU)

A new species of the catfish genus Centromochlus (Siluriformes: Auchenipteridae: Centromochlinae) from the upper rio Paraná basin, Brazil

Centromochlus comprises twelve species, distributed in the main inland watersheds of South America, including the Orinoco, Essequibo, coastal rivers of Suriname, Amazon, upper Paraná and São Francisco basins. The new species is described from the upper rio Paraná based on material collected in 1965 during the construction of the UHE Ilha Solteira, São Paulo, Brazil. The new species is easily distinguished from all congeners due to absence of adipose fin, a condition otherwise restricted to Gelanoglanis nanonocticolus, among centromochlin catfishes. The new species comprises small catfishes (adults ranging from 35 to 39 mm SL), in which modified anal fin of males is devoid of denticulations or spines, and most posterior rays reduced in length. In addition, Tatia simplex Mees is transferred to Centromochlus and its generic reassignment discussed.

Centromochlus abriga doze espécies, com registros para os principais sistemas fluviais de águas interiores da América do Sul, como o Orinoco, Essequibo, rios costeiros do Suriname, Amazonas, alto Paraná e São Francisco. A nova espécie é descrita do alto rio Paraná, a partir de material coletado em 1965 durante a construção da UHE Ilha Solteira, São Paulo, Brasil. A espécie nova distingue-se facilmente de todas as suas congêneres pela ausência de nadadeira adiposa, uma condição até então restrita à Gelanoglanis nanonocticolus, dentre os bagres centromochlíneos. A nova espécie inclui pequenos bagres (adultos entre 35 e 39 mm CP), nos quais a nadadeira anal modificada em machos é desprovida de dentículos ou espinhos, e com os raios mais posteriores de tamanho reduzido. Adicionalmente, Tatia simplex Mees é transferida para Centromochlus e é fornecida uma discussão acerca de seu posicionamento genérico.

A new miniature characid (Ostariophysi: Characiformes: Characidae), with phylogenetic position inferred from morphological and molecular data.

Erythrocharax altipinnis is described from the Serra do Cachimbo, Pará, Brazil. The new taxon is distinguished from all of the Characidae genera by having the pelvic bones firmly attached through the isquiatic processes; a nearly triangular hiatus in the musculature covering the anterior chamber of the swim bladder between the first and second pleural ribs (pseudotympanum); the pedunculate, notably expanded and distally compressed teeth in both jaws; circumorbital series represented by antorbital and four infraorbital bones with laterosensory canals not enclosed; a single tooth row in the premaxillary with the teeth perfectly aligned and similar in shape and cusp number; the first three branched dorsal-fin rays distinctly elongate in males; a bright red adipose and caudal fins in life; a conspicuous dark midlateral stripe extending from the opercle to the tip of the median caudal-fin rays; and by the absence of a humeral spot. The phylogenetic position of the new taxon is discussed using morphological and molecular datasets, with conflicting results of both approaches discussed. Additionally, a summarized discussion on the current problems in the Characidae taxonomy is presented and the principal biases in the morphological dataset are also discussed.