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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., (© 2024 Fisheries Society of the British Isles.)

Using integrative tools can be effective for species identification, especially in complex groups like Astyanax . Astyanax bimaculatus group is composed of six valid species, including A. lacustris . " A. altiparanae ", " A. asuncionensis ", and " A. jacuhiensis " are considered as junior synonyms of A. lacustris . Seeking to test the operational taxonomic unit (OTU) status of the junior synonyms of A. lacustris (" A. altiparanae ", " A. asuncionensis ", and " A. jacuhiensis "), we used analyses through mitochondrial DNA ( COI and Cytb ), cytogenetic markers (classical and molecular), and morphometry ("truss network"). Analysis of mitochondrial DNA sequences separated A. lacustris from the other synonymized species. The cytogenetic and morphometric analyses did not corroborate the synonymization and suggest that besides A. lacustris , the OTUs A. altiparanae , A. asuncionensis , and A. jacuhiensis are valid species. The analysis of different characters proposed by the integrative taxonomy used on the same individuals could provide greater reliability and minimize the underestimation of biodiversity.

The scattered distribution pattern of microsatellites is a challenging problem in fish cytogenetics. This type of array hinders the identification of useful patterns and the comparison between species, often resulting in over-limited interpretations that only label it as "scattered" or "widely distributed". However, several studies have shown that the distribution pattern of microsatellites is non-random. Thus, here we tested whether a scattered microsatellite could have distinct distribution patterns on homeologous chromosomes of closely related species. The clustered sites of 18S and 5S rDNA, U2 snRNA and H3/H4 histone genes were used as a guide to compare the (GATA)n microsatellite distribution pattern on the homeologous chromosomes of six Trachelyopterus species: T. coriaceus and Trachelyopterus aff. galeatus from the Araguaia River basin; T. striatulus, T. galeatus and T. porosus from the Amazonas River basin; and Trachelyopterus aff. coriaceus from the Paraguay River basin. Most species had similar patterns of the (GATA)n microsatellite in the histone genes and 5S rDNA carriers. However, we have found a chromosomal polymorphism of the (GATA)n sequence in the 18S rDNA carriers of Trachelyopterus galeatus, which is in Hard-Weinberg equilibrium and possibly originated through amplification events; and a chromosome polymorphism in Trachelyopterus aff. galeatus, which combined with an inversion polymorphism of the U2 snRNA in the same chromosome pair resulted in six possible cytotypes, which are in Hardy-Weinberg disequilibrium. Therefore, comparing the distribution pattern on homeologous chromosomes across the species, using gene clusters as a guide to identify it, seems to be an effective way to further the analysis of scattered microsatellites in fish cytogenetics., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Haerter et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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.

The aims of the study were: (i) evaluate the efficacy of using amphibian adipose tissue as a valid biomarker of water contaminated by pyriproxyfen; and (ii) verify the use of Lithobates catesbeianus in laboratory experiments as a potential bioindicator for bioaccumulation of pesticide in adipose tissue from amphibians. Adult frogs were exposed to different dilutions (0.002 g/L and 0.02 g/L) of Sumilarv® (pyriproxyfen) over 50 days. The average results of the fortified sample were 108%, indicating that the test method was effective. Adult frogs exposed to the standard dose recommended by the World Health Organization (WHO) showed bioaccumulation of pyriproxyfen in adipose tissue significantly higher than control animals. Lithobates catesbeianus proved to be an effective bioindicator and the adipose tissue was an efficient biomarker to bioaccumulation of pyriproxyfen. We conclude that high-performance liquid chromatography-mass spectrometry was effective measuring pyriproxyfen bioaccumulation in adult amphibians., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Farlowella is the second richest genus in Loricariinae, broadly distributed in freshwater streams and rivers of South America. In this article, we aimed to expand on the cytogenetic and molecular data available for two allopatric populations of Farlowella hahni . Both populations had diploid chromosome number 58, but with karyotype differences, indicative of chromosomal rearrangements. C-banding showed large heterochromatic blocks at telomeric regions in acrocentric chromosomes in both populations. Fluorescence in situ hybridization (FISH) revealed a single 18S rDNA site in both populations and a single 5S rDNA site for individuals from lower Paraná River basin (native region) and multiple 5S rDNA sites for individuals from upper Paraná River basin (non-native region). Mitochondrial sequence analyses did not separate the two F. hahni populations. The cytogenetic and molecular data obtained are relevant in a preliminary study and suggested the existence of cryptic diversity and the hypothesis that at least two Farlowella lineages may coexist in the Paraná basin.

Basic and molecular cytogenetic techniques were carried out in 3 Neotropical region populations of catfishes, two of Trachelyopterus galeatus (one from the marshlands of Paraguay River basin and another from Lago Catalão, Amazon River basin) and one of Trachelyopterus porosus, a sympatric population to T. galeatus from the Amazon River basin. This study aimed to describe and understand the structure and evolution of Trachelyopterus B chromosomes, mainly through physical mapping of repetitive elements. A diploid number of 58 chromosomes was found for all individuals, as well as the presence of B chromosomes. For T. porosus this is the first report of a supernumerary. The sympatric species of T. galeatus and T. porosus from Amazon River had 1-3 B chromosomes and T. galeatus from Paraguay River had 1-2 B chromosomes, all of them showed intra- and interindividual numerical variation. Two females of T. porosus exhibited a new variant B chromosome (B2), previously not seen in Auchenipteridae, which might have originated from B1 chromosomes. All B chromosomes were entirely heterochromatic. In contrast to all complement A and B2 chromosomes, in which the telomeric sequences were found in the telomeric regions, B1 chromosomes of all populations were totally marked by (TTAGGG)n probes. (GATA)n sequence sites were found through all complement A chromosomes, but B1 and B2 chromosomes exhibited only a clustered block in one of the chromosome arms. The most frequent B chromosomes (B1) in all populations/species, including those previously studied in Auchenipteridae catfishes, share the following characteristics: totally heterochromatic, small, metacentric, with accumulation of repetitive (TTAGGG)n sequences, and a low number of (GATA)n copies, which might suggest a common ancient origin in Trachelyopterus species/populations., (© 2021 S. Karger AG, Basel.)

This study provides new insight into the chromosomal diversification in Loricariinae. We analyzed nine species from different Brazilian hydrographic basins, using conventional and molecular cytogenetic methods, aiming to understand the karyotypic diversification, and contribute with cytotaxonomic markers in this group considered one of the most diverse of Loricariidae. Our results evidenced a high karyotypic variability in diploid number (2 n ) ranging from 2 n  = 54 ( Loricariichthys platymetopon and Loricariichthys anus ), 2 n  = 60 ( Rineloricaria reisi and Rineloricaria parva ), 2 n  = 62 ( Proloricaria prolixa ), 2 n  = 64 ( Loricaria cataphracta complex species), 2 n =  66 ( Sturisoma barbatum ), and 2 n  = 68 ( Pyxiloricaria menezesi ). Different patterns of 18S and 5S ribosomal DNA (rDNA) were also identified, while slight divergences in heterochromatin distribution were observed. This high variability is probably related with independent events of Robertsonian translocations, pericentric inversions, and different mechanisms of rDNA sites dispersion (nonreciprocal translocation and transposable element [TEs] co-localization). In addition, our study provides a set of efficient chromosomal markers for the characterization of all analyzed species, and certainly, in future analyzes, will contribute as a useful cytotaxonomic tool in groups where the traditional taxonomy based on morphological data are not sufficient to clarify their relationship.

Background: One of the biggest challenges in chromosome biology is to understand the occurrence and complex genetics of the extra, non-essential karyotype elements, commonly known as supernumerary or B chromosomes (Bs). The non-Mendelian inheritance and non-pairing abilities of B chromosomes make them an interesting model for genomics studies, thus bringing to bear different questions about their genetic composition, evolutionary survival, maintenance and functional role inside the cell. This study uncovers these phenomena in multiple species that we considered as representative organisms of both vertebrate and invertebrate models for B chromosome analysis., Results: We sequenced the genomes of three animal species including two fishes Astyanax mexicanus and Astyanax correntinus, and a grasshopper Abracris flavolineata, each with and without Bs, and identified their B-localized genes and repeat contents. We detected unique sequences occurring exclusively on Bs and discovered various evolutionary patterns of genomic rearrangements associated to Bs. In situ hybridization and quantitative polymerase chain reactions further validated our genomic approach confirming detection of sequences on Bs. The functional annotation of B sequences showed that the B chromosome comprises regions of gene fragments, novel genes, and intact genes, which encode a diverse set of functions related to important biological processes such as metabolism, morphogenesis, reproduction, transposition, recombination, cell cycle and chromosomes functions which might be important for their evolutionary success., Conclusions: This study reveals the genomic structure, composition and function of Bs, which provide new insights for theories of B chromosome evolution. The selfish behavior of Bs seems to be favored by gained genes/sequences.

Cytogenetic data showed a variation in diploid chromosome number in the genus Hyphessobrycon ranging from 2n = 46 to 52, and studies involving repetitive DNA sequences are scarce in representatives of this genus. The purpose of this paper was the chromosomal mapping of repetitive sequences (rDNA, histone genes, U snDNA and microsatellites) and investigation of the amplification of 5S rDNA clusters in the Hyphessobrycon eques genome. Two H. eques populations displayed 2n = 52 chromosomes, with the acrocentric pair No. 24 bearing Ag-NORs corresponding with CMA 3 +/DAPI-. FISH with a 18S rDNA probe identified the NORs on the short (p) arms of the acrocentric pairs Nos. 22 and 24. The 5S rDNA probe visualized signals on almost all chromosomes in genomes of individuals from both populations (40 signals); FISH with H3 histone probe identified two chromosome pairs, with the pericentromeric location of signals; FISH with a U2 snDNA probe identified one chromosome pair bearing signals, on the interstitial chromosomal region. The mononucleotide (A), dinucleotide (CA) and tetranucleotide (GATA) repeats were observed on the centromeric/pericentromeric and/or terminal positions of all chromosomes, while the trinucleotide (CAG) repeat showed signals on few chromosomes. Molecular analysis of 5S rDNA and non-transcribed spacers (NTS) showed microsatellites (GATA and A repeats) and a fragment of retrotransposon (SINE3/5S-Sauria) inside the sequences. This study expanded the available cytogenetic data for H. eques and demonstrated to the dispersion of the 5S rDNA sequences on almost all chromosomes.

Wertheimerinae is a small subfamily of thorny catfish composed of two species found in eastern Brazilian coastal drainages: Wertheimeria maculata and Kalyptodoras bahiensis . According to molecular phylogenetic analysis, Franciscodoras marmoratus an endemic species of the São Francisco River is also a member of this subfamily. Even though both phylogenetic approaches suggest that this group is one of the oldest lineages of the Doradidae, a disagreement remains about the constitution of Wertheimerinae. Hence, cytogenetic analysis is important to understand the karyotypic evolution of thorny catfish and can be a useful cytotaxonomic tool to clarify the relationships between these species. All Wertheimerinae species, and F. marmoratus here analyzed, shared 2n = 58 chromosomes, karyotypic formulas (24m+12sm +8st +14a), and nucleolus organizer region (NOR) pattern (terminal 18S rDNA sites on pair 22). Differences were noted in heterochromatin and 5S rDNA site distribution. The chromosomal markers here applied added to the molecular data, reinforcing that these three species actually represent a well-resolved taxonomic unit. Our results represent one more evidence of the ancient connectivity between eastern coastal drainages and São Francisco River, whose separation represented an important event for the allopatric speciation that produced the current forms of Wertheimerinae subfamily.

Hypostominae is the largest subfamily of Loricariidae, and is widely distributed throughout the Neotropic. In the present article, we analyze three Loricariidae species that were considered part of Hypostominae, from three different tribes, to discuss chromosome evolution in this fish group and to review the existent data for the subfamily. Rhinelepis aspera had 54 chromosomes (20m + 26sm+8st), whereas Pterygoplichthys ambrosettii and Megalancistrus parananus had 52 chromosomes, with 16m+24sm+8st+4a and 18m+24sm+10st, respectively. The karyological data were compared with existent phylogenetic hypotheses, indicating a common ancestor with 2n = 52 chromosomes for the Acanthicus, Hemiancistrus, and Peckoltia clades, as well as for Hypostomini. Shared recurrent characteristics of the tribes are discussed, as well as peculiarities of genera Ancistrus and Hypostomus. We propose that the occurrence of fragile sites demonstrated for Ancistrus facilitated chromosomal rearrangements that decreased the proportion of metacentric/submetacentric chromosomes and the diploid number in many species from this genus. Although Hypostominae is usually considered a subfamily with derived chromosome features, our revision shows that this is valid only for Hypostomini and Ancistrini, which have a divergent chromosome evolution from other tribes that seems to conserve plesiomorphic features.

Astyanax is a species-rich polyphyletic genus distributed between the southern United States and central Argentina. The genus contains groups of cryptic species, which are difficult to distinguish, and are sometimes identified wrongly. Basic and molecular cytogenetic analyses were run on Astyanax abramis and three junior synonyms of Astyanax lacustris: Astyanax altiparanae, from the upper Paraná River basin, Astyanax asuncionensis, from the lower Paraná basin, and Astyanax jacuhiensis, from the upper Uruguay River. These species all belong to the Astyanax bimaculatus group. All species presented 2n = 50 chromosomes and single nucleolar organizing regions (NORs). In A. altiparanae, the karyotype was 6m + 28sm+4st+12a and the NORs were present in pair 20, while A. jacuhiensis was 8m + 28sm+6st+8a, with NORs in pair 22, and A. asuncionensis was 8m + 24sm+6st+12a, with NORs in pair 20. A. abramis was 4m + 30sm+8st+8a with NORs in pair 22. Fluorescence in situ hybridization revealed single 5S rDNA cistrons in A. altiparanae and A. asuncionensis, and multiple (4) cistrons in A. abramis and A. jacuhiensis. Heterochromatin had a distinct distribution in each species, but was predominantly centromeric and interstitial proximal. In A. abramis and A. asuncionensis, the first acrocentric chromosome pair presented centromeric, telomeric, and interstitial-proximal heterochromatin in the long arm, which may represent the presence of homologous chromosomes in these species. While there are some cytogenetic similarities, differences in the location of 5S rDNA, distribution of heterochromatin, and karyotype formulae contribute to the differentiation of the study species, and support the identification of phylogenetically proximate groups in the "Astyanax clade."

Doradidae has been a target of phylogenetic studies over the last few years, but chromosomal information about the family is still scarce. Therefore, new cytogenetic data are provided herein and they are correlated with phylogenetic proposals to contribute to the knowledge of chromosomal evolution within doradids. Cytogenetic studies were performed on Trachydoras paraguayensis, Anadoras sp. "araguaia," Ossancora eigenmanni, Platydoras armatulus, and Rhinodoras dorbignyi. O. eigenmanni, P. armatulus, and R. dorbignyi had 2n = 58 chromosomes as found for most doradids, but T. paraguayensis and Anadoras sp. "araguaia" had 2n = 56 chromosomes, probably caused by a chromosomal reduction. There is a great maintenance of 2n = 58 verified in doradids, but karyotype formulas are diverse. Moreover, other markers (i.e., nucleolar organizer regions, heterochromatin distribution, and 5S and 18S rDNA) showed a great diversity among the analyzed species. Contrasting the variability in the chromosomal markers with the maintenance of diploid number, it is likely that inversions and translocations played an important role in chromosome differentiation in Doradidae. Herein, we created an integrative discussion linking cytogenetic data to phylogenetic proposals, based on morphological and genetic features, enabling us to identify possible cytogenetic traits, as well as probable chromosomal plesiomorphy and apomorphy of Doradidae species.

The karyotype and chromosomal characteristics of Apteronotus albifrons and Apteronotus caudimaculosus collected from populations of two different large Brazilian river basins were analyzed by conventional and molecular cytogenetic techniques, to contribute to the differentiation and identification of the species in this genus. The diploid chromosome number was 2n = 24 for A. albifrons, but with difference in the karyotype structure and fundamental number values between two populations under study. In A. caudimaculosus, the diploid chromosome number was 2n = 26, which was classified as 22 metacentric (m), 2 submetacentric (sm), and 2 acrocentric (a) chromosomes. Heterochromatins were preferentially located in pericentromeric regions for both species. However, there are more C-banded chromosomes in A. caudimaculosus than A. albifrons. The sites of 18S DNA as revealed by fluorescence in situ hybridization (FISH) in the karyotypes of both species corresponded to sites revealed by Ag impregnation, although some additional 18S rDNA sites were observed in the genome of A. caudimaculosus. FISH with 5S rDNA-probe revealed interstitial sites on the m pair No. 1 for individuals of both A. albifrons populations, and in pericentromeric regions on the long arm of pair Nos. 5 and 9 in those of A. caudimaculosus. The karyotypes of A. albifrons and A. caudimaculosus indicated a reduction of 2n resulting from chromosomal fusion, as could be hypothesized from the presence of an interstitial telomere sequence in two chromosome pairs in karyotype of A. caudimaculosus. Thus, the present study demonstrated species-specific cytogenetic markers of otherwise morphologically very similar species A. albifrons and A. caudimaculosus.

Karyotypes and other chromosomal markers as revealed by conventional and molecular cytogenetic protocols in four species of the catfish family Doradidae from the Araguaia-Tocantins river basin, namely Hassar wilderi, Leptodoras cataniae, Tenellus leporhinus and Tenellus trimaculatus were examined. All species had diploid chromosome number 2n = 58 and karyotypes dominated by biarmed chromosomes, simple NOR phenotype, that is, one chromosome pair bearing this site in terminal position, but some differences in karyotypes and distribution of constitutive heterochromatin, position of rDNA sites. Such characteristics appeared species-specific. A ZZ/ZW sex chromosome system was found in Tenellus trimaculatus, resulting likely from the amplification of the heterochromatin, followed by a paracentric inversion. Our results confirmed low karyotype differentiation observed until now among representatives of this endemic catfish family.

Corydoras Lacepède, 1803 is the most specious genus of Corydoradinae subfamily and many of its species are still unknown in relation to molecular cytogenetic markers. However, the diploid number and karyotypic formula were recorded for many species of this group. In current study, we provided the first cytogenetic information of Corydoras carlae Nijssen & Isbrücker, 1983, an endemic fish species from Iguassu River basin, Paraná State, Brazil. The individuals were collected in Florido River, a tributary of Iguassu River and analysed with respect to diploid number, heterochromatin distribution pattern, Ag-NORs and mapping of 5S and 18S ribosomal genes. The karyotype of this species comprises 46 chromosomes arranged in 22m+22sm+2st. The heterochromatin is distributed in centromeric and pericentromeric positions in most of the chromosomes, and also associated with NORs. The Ag-NORs were detected in the terminal position on the long arm of the metacentric pair 6. The double-FISH technique showed that 5S rDNA and 18S rDNA were co-localized in the terminal portion on the long arm of the metacentric pair 6. This condition of co-localization of ribosomal genes in Corydoras carlae seems to represent a marker for this species.

The fish species Erythrinus erythrinus belongs to the family Erythrinidae (order Characiformes, superorder Teleostei) and is considered a species complex because of the considerable differences between the karyotypes of analyzed populations. Whereas some populations present a sex chromosome system with male heterogamety, others do not show differentiated sex chromosomes. In this article, two novel karyotypes of E. erythrinus with the occurrence of male and female heterogamety are described, and a discussion of the stability of multiple sex chromosome systems is provided. A possible cause for sex chromosomes instability is that the Robertsonian rearrangements that originated the multiple systems did not prevent recombination with ancestral chromosomes, which also did not pass through a heterochromatinization process, the opposite of what usually happens with simple systems, especially of the ZZ/ZW or XX/XY type. It is suggested that multiple sex chromosome systems would not act as an effective postzygotic barrier, especially when there are hybridization zones between distinct karyomorphs that bear and that do not bear sex chromosome systems, allowing the generation of hybrids. This finding is important both for the comprehension of sex chromosomes evolution in fish and for conservation biology since the contact between populations with and without multiple sex chromosomes may compromise the regional biodiversity.

The karyotype and chromosomal characteristics of Trachydorasparaguayensis, a representative of the South American catfish family Doradidae, were analyzed by conventional (Giemsa staining, silver staining, C-banding) and molecular (FISH with rDNA and telomeric probes) cytogenetic techniques. The diploid chromosome number was 2n = 56, with 36 metacentric, 16 submetacentric, and 4 subtelocentric chromosomes in both sexes; however, a remarkable heteromorphism in pair 22 (submetacentric and metacentric elements) was detected in 6 individuals. Compared to other representatives of Doradidae which mostly have 58 chromosomes, the karyotype of T. paraguayensis suggests a reduction in 2n due to chromosomal fusion, as could be deduced from the presence of an interstitial telomere sequence in the submetacentric pair 19. Pale heterochromatic blocks were present in the terminal regions of some chromosomes, very similar to other species of Doradidae. The interstitial position of the NORs observed in the karyotype of T. paraguayensis differs from those reported for most Doradidae species, indicating that it is a derived character. FISH with 5S rDNA revealed 2 interstitial fluorescent signals in the submetacentric pair 22, and the polymorphism of these sites likely resulted from a pericentric inversion., (© 2016 S. Karger AG, Basel.)

The Iguazu river is a tributary of the left margin of the Paraná river, isolated from this basin about 22 million years ago with the appearance of the Iguazu Falls. The Iguazu river is characterized by high endemism due to two factors: its rugged topography and the old isolation caused by formation of the Iguazu Falls. This study analyzed cytogenetically a population of Glanidium ribeiroi collected in a region at the final stretch of this basin, by Giemsa staining, C-banding, impregnation by silver nitrate, and FISH with probes of 5S rDNA, 18S rDNA, telomeric sequence [TTAGGG]n, and [GATA]n repeats. The diploid number was equal to 58 chromosomes. The heterochromatin was present in the terminal region of almost all chromosomes. The Ag-NORs were simple and presented interstitially on the short arm of the submetacentric pair 14, which was confirmed by FISH with 18S rDNA probe. The 5S rDNA-FISH marked only the submetacentric pair 16 on the long arm in interstitial position. The FISH with [TTAGGG]n probe presented all telomeres labeled as expected, with an absence of Interstitial Telomeric Sequence (ITS). The repetitive [GATA]n sequence was dispersed throughout the genome, with preferential location in the terminal region of all chromosomes. The data obtained are discussed herein with other species of Auchenipteridae, and other previously analyzed populations of G. ribeiroi from the Iguazu river, verifying differences among these populations, which should be mainly related to the rugged topography of this basin.

Karyotypes and chromosomal characteristics of both minor and major rDNAs in four fish species known popularly as "lambaris", namely Astyanaxabramis (Jenyns, 1842), Astyanaxasuncionensis Géry, 1972, Astyanaxcorrentinus (Holmberg, 1891) and Astyanax sp. collected from downstream of the Iguassu Falls (Middle Paraná River basin), preservation area of the Iguassu National Park, were analyzed by conventional and molecular protocols. Astyanaxabramis had diploid chromosome number 2n=50 (4m+30sm+8st+8a) and single AgNORs (pair 22), Astyanaxasuncionensis had 2n=50 (8m+24sm+6st+12a) and single AgNORs (pair 20), Astyanax sp. had 2n=50 (4m+26sm+8st+12a) and single AgNORs (pair 25), and Astyanaxcorrentinus had 2n=36 (12m+16sm+2st+6a) and multiple AgNORs (pairs 12, 15, 16, 17). FISH with 18S rDNA showed a single site for Astyanaxabramis, Astyanaxasuncionensis and Astyanax sp. and multiple for Astyanaxcorrentinus (14 sites). FISH with 5S rDNA showed single 5S-bearing loci chromosome pair only for Astyanaxasuncionensis and multiple for Astyanaxabramis (four sites), Astyanaxcorrentinus (five sites) and Astyanax sp. (four sites). Distinct distribution patterns of heterochromatin were observed for karyotypes of all species, with the exception of the first acrocentric chromosome pair characterized by centromeric, interstitial-proximal and telomeric blocks of heterochromatin on the long arm, which may represent homeology between karyotypes of Astyanaxabramis and Astyanaxasuncionensis. Our study showed species-specific characteristics which can serve in diagnosis and differentiation between Astyanaxabramis and Astyanaxasuncionensis, considered cryptic species, as well as strengthening the occurrence of a species of Astyanax not yet described taxonomically. In addition, the data obtained from first cytogenetic studies in Astyanaxcorrentinus suggest a high similarity with Astyanaxschubarti Britski, 1964, suggesting that these species may belong to the same morphological group and that can be phylogenetically related.

The karyotype of the Ancistrini catfish Ancistrus taunayi was analyzed by conventional (Giemsa staining, AgNOR staining and C-banding) and molecular cytogenetic (5S and 18S rDNA-FISH) methods. The diploid chromosome number was 2n = 50 (22 metacentrics + 10 submetacentrics + 10 subtelocentrics + 8 acrocentrics) for both sexes. A single NOR-bearing acrocentric chromosome pair (No. 24) was detected after Ag-staining and 18S rDNA-FISH, while 5S rDNA was found only in the subtelocentric pair No. 21. Conspicuous GC-rich heterochromatin blocks corresponded to the NOR sites and were also observed in the distal regions of the acrocentric chromosome pairs Nos. 22 and 25. Chromosome pair No. 22 differed between males and females; in males, only a small interstitial block of GC-rich heterochromatin was present in both chromosomes, whereas in females, 2 blocks of GC-rich heterochromatin flanked a euchromatic region in one of the homologues, suggesting the occurrence of a ZZ/ZW sex chromosome system. Two mechanisms for the origin and evolution of this simple ZZ/ZW sex chromosome system in A. taunayi are proposed: (1) a paracentric inversion followed by amplification of the proximal heterochromatin and (2) amplification of the interstitial heterochromatin followed by a paracentric inversion. Although ZZ/ZW systems have already been described for other Ancistrus species, our results do not show the same pattern, suggesting an independent origin., (© 2015 S. Karger AG, Basel.)

A cytogenetic analysis was performed on four populations of Hypostomus strigaticeps from the Paraná River basin, Brazil. Two populations were collected from the large channel river at the Itaipu reservoir area and the other two were from the upper stretches of tributaries of the Paraná River. All populations showed 2n=72 chromosomes (12m+12sm+18st+30a), intra- and interpopulation 18S rDNA site polymorphisms (two to three acrocentric chromosome pairs), and multiple 5S rDNA sites in three chromosome pairs (4, 21, and 28). C-banding revealed heterochromatin located in the centromere and pericentromere regions of most chromosome; however, large heterochromatic blocks (CMA3(-)/DAPI(+)) on the long arm of acrocentric chromosomes identified intra- and interpopulation polymorphism. The amount and distribution of heterochromatin seem to be correlated to biogeographical characteristics of H. strigaticeps along the Paraná River. Morphometric results also showed diversity among the populations, suggesting phenotypic plasticity of this species. Evolutionary, taxonomy, and biogeographical approaches with regard to H. strigaticeps and interrelationships in Hypostomus are discussed.

Molecular genetic assessments that consider ecological information, in addition to endogamy levels, genetic diversity, and the genetic differentiation among species and populations, are particularly important for the conservation of biological diversity. Prime candidates for conservation genetic review are those subject to human use, including harvests for the ornamental fish trade. Colorful South American tetra, such as Moenkhausia oligolepis and M. forestii, are good examples of fish species that are widely collected and exported worldwide. This study aimed to evaluate the population-specific characteristics of M. oligolepis and M. forestii by comparing morphometric and molecular analyses based on ISSR markers, to provide information that would facilitate the sustainable management of these 2 species. Seventy-two specimens were collected from the Araguaia-Tocantins and Paraguay River Basins in Brazil. All specimens were measured and analyzed using ISSR markers. Population-exclusive bands were found among the 86 detected bands, while morphometric clusters reflected the geographical distribution of individuals. Correlated genetic and morphological variation supported the presence of 3 distinct groups from tributaries of the Araguaia and Mortes Rivers. Using the same techniques, all M. oligolepis populations were isolated from M. forestii. This study on Moenkhausia presents an interesting example that could be used to construct a framework of South American ichthyodiversity, and reinforces the necessity of habitat conservation to prevent the loss of biological diversity.

Erythrinidae include Neotropical teleost fish that are widely distributed in South America. Hoplias Gill, 1903 include two large groups: H. malabaricus Bloch, 1794 and H. lacerdae Miranda Ribeiro, 1908. Hoplias malabaricus is characterized by remarkable karyotype diversity, with some karyomorphs widely distributed geographically while others are more restricted to certain river basins. Cytogenetic analyzes were performed in a population of Hoplias malabaricus from the Wildlife Refuge of Campos de Palmas, the Iguaçu River basin. The specimens showed diploid number of 42 chromosomes (24m+18sm) without differentiated sex chromosomes system. The impregnation by silver nitrate showed multiple AgNORs. Seven pairs (4, 7, 10, 13, 16, 20 and 21) carrying 18S rDNA were detected by FISH. Heterochromatin was verified in the centromeric and pericentromeric region of most chromosomes and the terminal region of some pairs. FISH with 5S rDNA probes showed two chromosome pairs carrying these sites in the interstitial region (8 and 14). The data obtained in this study are similar to those found for two other populations of H. malabaricus already studied in the basin of the Iguaçu River, confirming the hypothesis that this species is natural, not having been introduced, as well as having an intrinsic characteristic, such as the largest number of sites of 18S rDNA.

Representing a reflection of anthropic activity, the level of xenobiotic compounds in aquatic ecosystems has increased in recent years, bringing severe damage to the environment. The present work reports the occurrence of malformation in gonads of Physalaemus cuvieri individuals from a population of Atlantic Forest in Southern Brazil. Twenty male specimens were collected, which had their testicles removed, immersed in Karnovsky fixative solution, included in historesin for 2 μm cuts and stained with Hematoxylin-eosin. Four specimens showed intersexual gonads condition along with the presence of sperm and oocytes. In order to test a possible contamination of water, 2L were collected from the water body to check organochlorine, organophosphate and carbamate compounds. The analysis of water showed the presence of agrotoxic Dieldrin in a concentration of 0.05 μg/L, representing a concentration above the recommended reference. This agrotoxic, in addition to acting as endocrine disrupter and commercially prohibited, has quite persistent residual effects, and may be responsible for the high frequency of P. cuvieri with intersexual gonads, which in the long term can represent a risk for this population due to the potential impact on its effective reproductive ability., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Harttia is a genus in the subfamily Loricariinae that accommodates fishes popularly known as armored catfishes. They show extensive karyotypic diversity regarding interspecific numerical/structural variation of the karyotypes, with the presence of the XX/XY1Y2 multiple sex chromosome system, as found in H. carvalhoi. In this context, this study aimed to characterize Harttia punctata chromosomally, for the first time, and to infer the rearrangements that originated the X1X1X2X2/X1X2Y multiple sex chromosome system present in this species. The data obtained in this study, with classical (Giemsa, C-banding and AgNORs) and molecular methodologies (fluorescence in situ hybridization) and chromosome microdissection, indicated that a translocation between distinct acrocentric chromosomes bearing rRNA genes, accompanied by deletions in both chromosomes, might have originated the neo-Y chromosome in this species. The data also suggest that the multiple sex chromosome systems present in H. carvalhoi and H. punctata had an independent origin, evidencing the recurrence of chromosome alterations in species from this genus.

Hypostomus is a diverse group with unclear aspects regarding its biology, including the mechanisms that led to chromosome diversification within the group. Fluorescence in situ hybridization (FISH) with 5S and 18S rDNA probes was performed on ten Hypostomini species. Hypostomus faveolus, H. cochliodon, H. albopunctatus, H. aff. paulinus, and H. topavae had only one chromosome pair with 18S rDNA sites, while H. ancistroides, H. commersoni, H. hermanni, H. regani, and H. strigaticeps had multiple 18S rDNA sites. Regarding the 5S rDNA genes, H. ancistroides, H. regani, H. albopunctatus, H. aff. paulinus, and H. topavae had 5S rDNA sites on only one chromosome pair and H. faveolus, H. cochliodon, H. commersoni, H. hermanni, and H. strigaticeps had multiple 5S rDNA sites. Most species had 18S rDNA sites in the telomeric region of the chromosomes. All species but H. cochliodon had 5S rDNA in the centromeric/pericentromeric region of one metacentric pair. Obtained results are discussed based on existent phylogenies for the genus, with comments on possible dispersion mechanisms to justify the variability of the rDNA sites in Hypostomus.

Auchenipteridae is divided in two subfamilies, Centromochlinae and Auchenipterinae. Centromochlinae has 31 valid species, from which 13 are included in the genus Tatia Miranda Ribeiro, 1911. Among these, Tatia jaracatia Pavanelli & Bifi, 2009 and Tatia neivai (Ihering, 1930) are the only two representative species from the Paraná-Paraguay basins. This study aimed to analyze cytogenetically these two species and thus provide the first chromosomal data for the genus. Although Tatia jaracatia and Tatia neivai presented 2n=58 chromosomes, some differences were observed in the karyotypic formula. The heterochromatin was dispersed in the centromeric and terminal regions of most chromosomes of Tatia jaracatia, and only in the terminal region of most chromosomes of Tatia neivai. The AgNORs were detected in the subtelocentric pair 28 for both species, which was confirmed by FISH with 18S rDNA probe. The 5S rDNA sites were detected in four chromosome pairs in Tatia jaracatia and three chromosome pairs in Tatia neivai. Both species of Tatia presented great chromosomal similarities among themselves; however, when compared to other species of Auchenipteridae, it was possible to identify some differences in the karyotype macrostructure, in the heterochromatin distribution pattern and in the number and position of 5S rDNA sites, which until now seems to be intrinsic to the genus Tatia.

The greatest diversity of anurans in the world is in Brazil and one of the major challenges is to reconcile the accelerated economic development with strategies that aim to maintain this diversity in forest fragments, often representing ESUs of some biomes. This study aimed to obtain data that will support conservation projects through the pioneering use of ISSR analysis in Neotropical anurans, estimating the intra- and interpopulation genetic diversity of four populations of P. cuvieri (Paraná and São Paulo regions). Of the 65 loci scored 58 were polymorphic, with 0.797 intrapopulation variation and 0.203 interpopulation variation. The index of interpopulation genetic differentiation (FST) proved to be high among the population of Marmeleiro-PR and the three populations of SP (FST > 0.288); genetic dissimilarity was related to the geographical distance. The ISSR proved to be efficient and useful molecular markers in comparison with other markers most widely used for preliminary diagnosis of genetic diversity in populations of amphibians, and could be applied as a tool for future conservation projects, since they could identify potential ESUs and influence decisions on the preservation of fragments.

This study analyzed sequences of the control region of mitochondrial DNA (D-loop) in three populations of Parauchenipterus galeatus from the basins of the Paraná, São Francisco, and Piumhi rivers, of which the last river being a region that suffered transposition river. A fragment of 850 base pairs was obtained with a total of 65 polymorphic sites. The data discuss aspects related to the genetic distance between the populations through the phylogenetic reconstruction methods (neighbor-joining, maximum-likelihood, and Bayesian analysis). Moreover, the data suggest that the Piumhi River population (transposition region) has recently gone through a significant bottleneck effect, which must be directly related to the anthropic action that occurred in this region, since the drainage the old existing swamp was necessary for the construction of the transposition channel potentially leading this population in to the current lack of genetic diversity.

The Neotropical fish Heptapterus mustelinus, collected in the Pindorama stream of the upper Paraná River basin (Brazil), was studied cytogenetically, verifying 54 chromosomes (26m + 18m + 4st + 6a). This diploid number has not been reported among the Heptapteridae that have been studied to date. Unlike most species of the family, there were multiple Ag-nucleolar organizer regions (NOR) and heterochromatin present in the centromeric region of most of the chromosomes of the complement, being Chromomycin A(3) (CMA(3)(+))/4',6-diamidino-2-phenylindole (DAPI(-)) in nine chromosomal pairs, besides the one that coincides with the Ag-NORs. The data presented in this work reveal a different path in the karyotypic evolution of H. mustelinus when compared to the others Heptapteridae genera., (© 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.)

This study presents an adaptation of current methodologies for preparing mitotic chromosomes from fishes, optimized for use in the field. The high-quality preparations obtained using this modified methodology is suitable for subsequent chromosomal analysis. Importantly, this method is particularly useful when specimen collection sites are far from research laboratories or when researchers are working with highly sensitive species that do not survive long outside of their natural habitats., (© 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.)

The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme--GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species., Competing Interests: The authors have declared that no competing interests exist.

Neotropical fishes have a low rate of chromosome differentiation between sexes. The present study characterizes the first meiotic analysis of sex chromosomes in the order Gymnotiformes. Gymnotus pantanal - females had 40 chromosomes (14m/sm, 26st/a) and males had 39 chromosomes (15m/sm, 24st/a), with a fundamental number of 54 - showed a multiple sexual determination chromosome system of the type X(1)X(1)X(2)X(2)/X(1)X(2)Y. The heterochromatin is restricted to centromeres of all chromosomes of the karyotype. The meiotic behavior of sex chromosomes involved in this system in males is from a trivalent totally pared in the pachytene stage, with a high degree of similarity. The cells of metaphase II exhibit 19 and 20 chromosomes, normal disjunction of sex chromosomes and the formation of balanced gametes with 18 + Y and 18 + X(1)X(2) chromosomes, respectively. The small amount of heterochromatin and repetitive DNA involved in this system and the high degree of chromosome similarity indicated a recent origin of the X(1)X(1)X(2)X(2)/X(1)X(2)Y system in G. pantanal and suggests the existence of a simple ancestral system with morphologically undifferentiated chromosomes., (Copyright © 2011 S. Karger AG, Basel.)

Physical mapping of 5S rDNA in 2 species of knifefishes, Gymnotuspantanal and G. paraguensis (Gymnotiformes), was performed using fluorescence in situ hybridization with a 5S rDNA probe. The 5S rDNA PCR product from the genomes of both species was also sequenced and aligned to determine non-transcribed spacer sequences (NTS). Both species under study had different patterns of 5S rDNA gene cluster distribution. While in the karyotype of G. pantanal two 5S rDNA-bearing pairs were observed, the karyotype of G. paraguensis possessed as many as 19 such pairs. Such multiplication of 5S rDNA gene clusters might be caused by the involvement of transposable elements because the NTS of G. paraguensis was 400 bp long with high identity (90%) with a mobile transposable element called Tc1-like transposon, described from the cyprinid fish Labeo rohita., (Copyright © 2011 S. Karger AG, Basel.)

Conventional and molecular chromosomal analyses were carried out on three populations of Apareiodon ibitiensis sampled from the hydrographic basins of the São Francisco River and Upper Paraná River (Brazil). The results reveal a conserved diploid number (2n = 54 chromosomes), a karyotype formula consisting of 50 m-sm + 4st and a ZZ/ZW sex chromosome system that has not been previously identified for the species. C-banding analysis with propidium iodide staining revealed centromeric and terminal bands located in the chromosomes of the specimens from the three populations and allowed the identification of heteromorphism of heterochromatin regions in the Z and W chromosomes. The number of 18S sites located through fluorescent in situ hybridization (FISH) varied between the populations of the São Francisco and Upper Paraná Rivers. The location of 5S rDNA sites proved comparable in one pair of metacentric chromosomes. Thus, the present study proposes a ZZ/ZW sex chromosome system for A. ibitiensis among the Parodontidae, and a hypothesis is presented regarding possible W chromosome differentiation stages in this species through DNA accumulation, showing geographical variations for this characteristic, possibly as a consequence of geographical reproductive isolation.

B chromosomes are considered additional and non-essential; they likely originate from A chromosomes and follow a distinct evolution. In fish, approximately half of the Neotropical species with B chromosomes are Characiformes and 35% are Siluriformes. There has been no report of B chromosomes in Auchenipteridae until this moment. B chromosomes found in a population of Parauchenipterus galeatus from the São Francisco River basin in the state of Minas Gerais (Brazil) were small, metacentric, totally heterochromatic and exhibited intra-individual and inter-individual variation. The diploid number was 58 chromosomes (22 metacentric, 16 submetacentric, 12 subtelocentric and 8 acrocentric). The nucleolar organizing regions were simple and the heterochromatin intercalated in the ribosomal sites, characterized by CMA(3) and DAPI fluorochromes, was of a GC-rich constitution. The 5S rDNA genes were located in an intercalary position in only one chromosome pair. An hypothesis about the origin of the B chromosomes in P. galeatus and a review on B chromosomes in catfish are also presented in this study.

Karyotype, sex chromosome system and cytogenetics characteristics of an unidentified species of the genus Apareiodon originating from Piquiri River (Paraná State, Brazil) were investigated using differential staining techniques (C-banding and Ag-staining) and fluorescent in situ hybridization (FISH) with 5S and 18S rDNA probes. The diploid chromosome number was 2n = 54 with 25 pairs of meta- (m) to submetacentric (sm) and 2 pairs of subtelocentric (st) chromosomes. The major ribosomal rDNA sites as revealed by Ag-staining and FISH with 18S rDNA probe were found in distal region of longer arm of st chromosome pair 26, while minor 5S sites were observed in the interstitial sites on chromosome pairs 2 (smaller cluster) and 7 (larger one). The C-positive heterochromatin had pericentromeric and telomeric distribution. The heteromorphic sex chromosome system consisted of male ZZ (pair 21) and female middle-sized m/st Z/W chromosomes. The pericentric inversion of heterochromatinized short arm of ancestral Z followed by multiplication of heterochromatin segments is hypothesized for origin of W chromosome. The observed karyotype and chromosomal markers corresponded to those found in other species of the genus.

The genus Erythrinus belongs to the family Erythrinidae, a neotropical fish group. This genus contains only two described species, Erythrinus erythrinus being the most widely distributed in South America. Six samples of this species from five distinct Brazilian localities and one from Argentina were studied cytogenetically. Four groups were identified on the basis of their chromosomal features. Group A comprises three samples, all with 2n = 54 chromosomes, a very similar karyotypic structure, and the absence of chromosome differentiation between males and females. One sample bears up to four supernumerary microchromosomes, which look like 'double minute chromosomes' in appearance. Groups B-D comprise the three remaining samples, all sharing an X(1)X(1)X(2)X(2)/X(1)X(2)Y sex chromosome system. Group B shows 2n = 54/53 chromosomes in females and males, respectively, and also shows up to three supernumerary microchromosomes. Groups C and D show 2n=52/51 chromosomes in females and males, respectively, but differ in the number of metacentric, subtelocentric, and acrocentric chromosomes. In these three groups (B-D), the Y is a metacentric chromosome clearly identified as the largest in the complement. The present results offer clear evidence that local samples of E. erythrinus retain exclusive and fixed chromosomal features, indicating that this species may represent a species complex.