Topotype-Based Chromosomal Diversity among Five Species of Freshwater Armored Catfishes in the Hypostomus auroguttatus Supergroup (Actinopterygii: Siluriformes).

Freshwater catfishes from the genus Hypostomus have been models for several cytogenetic studies, due to their intense variability in diploid number, chromosome morphology, and the distribution of repetitive DNAs. Taking into consideration the taxonomic complexity inherent to this group, the present study aims to describe the karyotypes of five species of Hypostomus collected in their type localities: Hypostomus albopunctatus (Regan, 1908), Hypostomus hermanni (Ihering, 1905), Hypostomus iheringii (Regan, 1908), and Hypostomus paulinus (Ihering, 1905) from the Piracicaba River (the Upper Paraná River Basin); and Hypostomus mutucae Knaack, 1999 from the Claro River (the Upper Paraguay River Basin). Our results evidenced a great inter-specific diploid-number variation: 2n = 72 (H. hermanni); 2n = 74 (H. albopunctatus); 2n = 76 (H. paulinus); 2n = 80 (H. iheringii); and 2n = 82 (H. mutucae), which reflects the important role of Robertsonian rearrangements in the karyotypic differentiation among these species. The distribution of heterochromatin also varied considerably among species, making it possible to distinguish each analyzed species, as well as to detect microstructural variations among populations of the same species. These data can support taxonomic revisions when further associated with molecular markers and morphological analyses to delimit, more consistently, the taxonomic status of these Hypostomus species, which have a complex taxonomic diagnosis history.

Chromosome Mapping of 18S Ribosomal RNA Genes in Eleven Hypostomus Species (Siluriformes, Loricariidae): Diversity Analysis of the Sites.

We investigated the chromosomal distribution of 18S ribosomal DNA (rDNA) in different populations of 11 species of Hypostomus collected in important Brazilian basins, namely South Atlantic, Upper Paraná, and Paraguay applying the fluorescence in situ hybridization (FISH). Hypostomus cochliodon, Hypostomus commersoni, Hypostomus hermanni, Hypostomus regani, Hypostomus albopunctatus, Hypostomus paulinus, Hypostomus aff. paulinus, Hypostomus iheringii, and Hypostomus mutucae presented multiple 18S rDNA sites while Hypostomus strigaticeps and Hypostomus nigromaculatus exhibited a single pair of chromosomes with 18S rDNA sites. The studied species presented variations in the number and position of these sites. The results accomplished were similar to those obtained by the analysis of AgNORs, revealing the same interspecific variability. Each species exhibited distinctive patterns of AgNOR and 18S rDNA distribution, which can be considered cytogenetic markers in each species of the genus and help improve the discussions on the phylogeny of the group.

Sympatric occurrence of four cytotypes and one extra chromosome in Bryconamericus ecai (Characidae): 18S rDNA polymorphism and heterochromatin composition.

In the present study, specimens of Bryconamericus ecai collected from the Forquetinha River/RS, were cytogenetically analyzed, disclosing a wide karyotypic diversity in this species. All individuals had 2n = 50, with different karyotypic formulae, resulting in four cytotypes and one B macrochromosome observed in cytotype III. Heterochromatin was distributed in the pericentromeric region of most chromosomes on the four cytotypes and also on a chromosome pair with interstitial markings in cytotype IV. Staining with CMA(3) and DAPI fluorochromes revealed a C-band region rich in AT base pairs in cytotypes I, II and III, and a pair with GC-rich heterochromatin in cytotypes II and III. Cytotype IV presented CMA(3) and DAPI positive heterochromatin. Silver nitrate impregnation, in situ hybridization, and fluorochrome staining showed a multiple system of AgNORs, 18S rDNA and CMA(3) sites in cytotypes I, III and IV, with both inter-and intraindividual variability in the number and location of these sites. Cytotype II had only one pair of NORs coincident with the 18S rDNA and CMA(3) sites, indicating a simple system. The chromosomal polymorphism observed among the specimens of B. ecai added to the literature data show that chromosomal rearrangements, especially pericentric inversions, play an important role in the karyotypic evolution of this group of fish. It can also be implied that more than one species of Bryconamericus is probably occurring, living in sympatry in the Forquetinha River/RS.

Cytogenetic characterization of four species of the genus Hypostomus Lacépède, 1803 (Siluriformes, Loricariidae) with comments on its chromosomal diversity.

Cytogenetic analyses were performed on fishes of the genus Hypostomus (Hypostomus ancistroides (Ihering, 1911), Hypostomus strigaticeps (Regan, 1908), Hypostomus regani (Ihering, 1905), and Hypostomus paulinus (Ihering, 1905)) from the seven tributaries of the Paranapanema River Basin (Brazil) by means of different staining techniques (C-, Ag-, CMA3- and DAPI-banding) and fluorescence in situ hybridization (FISH) to detect 18S rDNA sites. All species showed different diploid numbers: 2n=68 (10m+26sm+32st-a) in Hypostomus ancistroides, 2n=72 (10m+16sm+46st-a) in Hypostomus strigaticeps, 2n=72 (10m+18sm+44st-a) in Hypostomus regani and 2n=76 (6m+16sm+54st-a) in Hypostomus paulinus. Ag-staining and FISH revealed various numbers and locations of NORs in the group. NORs were usually located terminally on the subtelocentric/acrocentric chromosomes: on the long arm in Hypostomus strigaticeps (2 to 4) and Hypostomus paulinus (2); and on the short arm in Hypostomus ancistroides (2 to 8) and Hypostomus regani (2 to 4). Conspicuous differences in heterochromatin distribution and composition were found among the species, terminally located in some st-a chromosomes in Hypostomus ancistroides, Hypostomus strigaticeps, and Hypostomus paulinus, and interstitially dispersed in most st-a chromosomes, in Hypostomus regani. The fluorochrome staining indicated that different classes of GC and/or AT-rich repetitive DNA evolved in this group. Our results indicate that chromosomal rearrangements and heterochromatin base-pair composition were significant events during the course of differentiation of this group. These features emerge as an excellent cytotaxonomic marker, providing a better understanding of the evolutionary mechanisms underlying the chromosomal diversity in Hypostomus species.

Coexistence of cytotypes and chromosomal mosaicism in Hoplias malabaricus (Characiformes, Erythrinidae).

Karyotypes of seventeen Hoplias malabaricus specimens, collected in the fish culture station of UNOPAR (University of Northern Paraná), were analyzed. The station is in the Claro River system in the Tibagi River basin. Two distinct and coexistent karyotype forms (cytotypes) were identified, comprising either 42 chromosomes (cytotype A) or 40 chromosomes (cytotype C), both presenting metacentric and submetacentric chromosomes. In two specimens, one male and one female, it was not possible to characterize a modal diploid number because different cell lines were observed, with a predominance of 2n=41 and 2n=42 chromosomes at a frequency of 38.24% and 41.12%, respectively. The karyotype with 2n=41 showed some putative monosomic and trisomic chromosomes, while the karyotype with 2n=42 showed 21 chromosomal pairs, similar to cytotype A. RAPD analysis showed that these two specimens have the same band pro file of cytotype A (Nei's genetic identity=92%), discarding a possible hybridization between both cytotypes and supporting the mosaicism hypothesis. These findings corroborate the isolation between cytotypes A and C.

Cytogenetic analysis of Astyanax laticeps (Cope, 1894) (Ostariophysi: Characidae) from the laguna dos Patos system

The genus Astyanax is prominent among Characiformes, due to the large number of species found and its wide geographic distribution. In this work, Astyanax laticeps specimens from the laguna dos Patos system were cytogenetically analyzed. A diploid number of 2n = 50 chromosomes distributed into 6m+16sm+16st+12a (FN = 88) was found, without differences between males and females. A few small heterochromatin blocks were observed, besides three more conspicuous C-bands, corresponding to NORs, as confirmed by silver nitrate and CMA3 staining, FISH, and DAPI negative staining. These regions were located in a medium-sized subtelocentric and in a large subtelocentric chromosomal pair, probably because of a deletion of this region in one homologous chromosome, or due to a transposition event between them.(AU)

O gênero Astyanax é destacado entre os Characiformes, pelo grande número de espécies encontradas e a ampla distribuição geográfica. Neste trabalho, foram analisados citogeneticamente espécimes de Astyanax laticeps do sistema da laguna do Patos. O número diplóide observado foi de 2n = 50 cromossomos distribuídos em 6m+16sm+16st+12a (NF= 88), sem diferenças entre machos e fêmeas. Foram observados poucos blocos de heterocromatina, além de três bandas-C mais conspícuas, correspondentes às NORs, confirmado pela coloração com nitrado de prata,CMA3, FISH, e coloração negativa ao DAPI. Estas regiões foram localizadas em um cromossomo subtelocêntrico de tamanho médio e em um par subtelocêntrico grande, provavelmente devido a deleção desta região em um dos cromossomos homólogos, ou por eventos de transposição entre eles.(AU)

Coexistence of cytotypes and chromosomal mosaicism in Hoplias malabaricus (Characiformes, Erythrinidae)

Karyotypes of seventeen Hoplias malabaricus specimens, collected in the fish culture station of UNOPAR (University of Northern Paraná), were analyzed. The station is in the Claro River system in the Tibagi River basin. Two distinct and coexistent karyotype forms (cytotypes) were identified, comprising either 42 chromosomes (cytotype A) or 40 chromosomes (cytotype C), both presenting metacentric and submetacentric chromosomes. In two specimens, one male and one female, it was not possible to characterize a modal diploid number because different cell lines were observed, with a predominance of 2n=41 and 2n=42 chromosomes at a frequency of 38.24 percent and 41.12 percent, respectively. The karyotype with 2n=41 showed some putative monosomic and trisomic chromosomes, while the karyotype with 2n=42 showed 21 chromosomal pairs, similar to cytotype A. RAPD analysis showed that these two specimens have the same band pro file of cytotype A (Nei's genetic identity=92 percent), discarding a possible hybridization between both cytotypes and supporting the mosaicism hypothesis. These findings corroborate the isolation between cytotypes A and C.

Cytogenetic characterization of Hypostomus nigromaculatus (Siluriformes: Loricariidae)

Hypostomus is the most speciose genus in the family Loricariidae, with approximately 120 species. These fish show a wide morphological and color variation, which hinders the identification of species, mainly of widely distributed representatives. The aim of this study was to contribute to the current knowledge on cytogenetic features of Hypostomus nigromaculatus. Three specimens of H. nigromaculatus, collected in two tributaries of rio Tibagi, Paraná, and in Cachoeira de Emas, rio Mogi-Guaçu, São Paulo, the latter being the type locality of H. nigromaculatus, were studied. Chromosomal preparations were submitted to Giemsa staining, silver nitrate impregnation, C-banding and CMA3 and DAPI fluorochromes staining. All samples presented 2n = 76, but the rio Mogi-Guaçu sample differed from those from tributaries of rio Tibagi in relation to karyotype formulae, distribution and composition of heterochromatin, and NOR location. The silver nitrate staining revealed the presence of multiple Ag-NORs for all samples, but with differences on the location on chromosomes. CMA3 staining reveled bright signals equivalent to NOR-bearing chromosomal segments; such sites were characterized by negative, i.e. unstained, marks after DAPI staining. The pattern of heterochromatin distribution was distinctive among samples from rio Mogi-Guaçu and tributaries of rio Tibagi. The differences observed between the sample from rio Mogi-Guaçu and the ones from tributaries of rio Tibagi allow us to suggest that these samples are presently isolated. Further analyses are necessary to ascertain whether such isolation refers to distinct populations or characterizes true different species.

O gênero Hypostomus é um dos mais especiosos na família Loricariidae, tendo aproximadamente 120 espécies. Apresenta uma ampla diversidade quanto ao padrão de coloração e morfologia, o que dificulta a identificação de determinadas espécies, principalmente aquelas com ampla distribuição geográfica. Para isso os dados obtidos neste trabalho contribuem para os estudos citogenéticos de Hypostomus nigromaculatus. Foram analisados três exemplares de H. nigromaculatus de afluentes do rio Tibagi, Paraná e Cachoeira de Emas, rio Mogi-Guaçu, São Paulo, sendo esta última, a localidade tipo de H. nigromaculatus. Os cromossomos foram submetidos à coloração convencional (Giemsa), impregnação por nitrato de prata, bandamento-C e coloração com os fluorocromos CMA3 e DAPI. Todos os exemplares apresentaram 2n = 76, no entanto com diferença quanto às fórmulas cariotípicas, distribuição e composição da heterocromatina. O nitrato de prata detectou RONs múltiplas para as amostras, porém com diferenças quanto à localização nos cromossomos. A coloração com fluorocromo CMA3 foi correspondente aos cromossomos Ag-RONs, na coloração com DAPI foram observadas bandas negativas, ou seja, não coradas. O padrão de distribuição da heterocromatina foi diferente para as amostras do rio Mogi-Guaçu e dos tributários do rio Tibagi. As diferenças observadas entre as amostras de rio Mogi-Guaçu e afluentes do rio Tibagi permitem-nos sugerir que essas amostras estejam atualmente isoladas. Outras análises são necessárias para determinar se as amostras analisadas são populações distintas ou trata-se de espécies diferentes.

Cytogenetic studies in three species of the genus Oligosarcus

Karyotypic studies were carried out on three species of the genus Oligosarcus: O. paranensis, O. pintoi, and O. longisrostris. All of them showed a diploid number of 50 chromosomes and the same karyotypic formulae (4M + 10SM + 16ST + 20A). Silver nitrate staining revealed single NORs in O. longirostris and multiple NORs in O. paranensis and O. pintoi. Heterochromatin seemed to play an important role in the chromosomal diversification of these species. Based on cytogenetical data, speciation hypotheses within this group were proposed, reinforcing the importance of chromosomal studies for a better understanding of evolution within the genus Oligosarcus, as well as within the family Characidae.

No presente trabalho foram analisadas três espécies do gênero Oligosarcus: O. paranensis, O. pintoi e O. longirostris. As três espécies possuem número diplóide de 50 cromossomos e mesma fórmula cariotípica (4M + 10SM + 16ST + 20A). A impregnação por nitrato de prata revelou RONs simples para O. longirostris e RONs múltiplas para O. paranensis e O. pintoi. A heterocromatina desempenha um importante papel na caracterização das espécies. Hipóteses sobre a especiação do grupo são apresentadas com base nos dados citogenéticos obtidos neste trabalho, reforçando a importância destes estudos para uma melhor compreensão da evolução tanto do gênero Oligosarcus quanto da família Characidae.