Unrevealing the Karyotypic Evolution and Cytotaxonomy of Armored Catfishes (Loricariinae) with Emphasis in Sturisoma, Loricariichthys, Loricaria, Proloricaria, Pyxiloricaria, and Rineloricaria.

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 (2n) ranging from 2n = 54 (Loricariichthys platymetopon and Loricariichthys anus), 2n = 60 (Rineloricaria reisi and Rineloricaria parva), 2n = 62 (Proloricaria prolixa), 2n = 64 (Loricaria cataphracta complex species), 2n = 66 (Sturisoma barbatum), and 2n = 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.

Cytogenetic Trends in Two Families of the Neotropical Catfishes: Heptapteridae and Pseudopimelodidae (Siluriformes).

Several neotropical Siluriformes groups suffered important taxonomic revisions based on the evaluation of morphological and molecular characteristics that allow the construction of new phylogenetic hypothesis. In the present study were cytogenetically analyzed six species belonging to Heptapteridae (Cetopsorhamdia iheringi, Phenacorhamdia tenebrosa, Rhamdella eriarcha, Pimelodella meeki, Pimelodella australis, Heptapterus mustelinus) and two to Pseudopimelodidae families (Microglanis cottoides and Microglanis cibelae) by means of differential staining techniques to describe more precisely cytogenetic similarities and differences. The diploid number of R. eriarcha with 2n = 58 and M. cibelae with 2n = 56 were reported for the first time. Also, the lowest chromosome number (2n = 48) for P. tenebrosa was described. The chromosome-banding techniques for to put in evidence nucleolar organizers impregnated by silver nitrate ([AgNORs], chromomycin A3 [CMA3], and rDNA 18S) showed for all studied species conserved patterns, characteristic for each family. The rDNA 5S showed high variability among species or populations of both families, these regions could be simple or multiple, syntenic, or not with rDNA18S. The chromosome markers showed that both families are related not only from a morphologic point of view but also by their karyotypic characteristics, however, some of the present cytogenetic results evidence the importance of new morphologic, molecular, and phylogenetic studies to improve the knowledge of these fish groups.

First chromosome data on Steindachneridion doceanum (Siluriformes: Pimelodidae): a critically endangered catfish endemic of the Doce River basin, Brazil

The present report represents the first cytogenetic description of Steindachneridion doceanum, great catfish which is currently at high extinction risk and it is listed as threatened on the red list of the Brazilian Ministry of the Environment, also are suggested karyotype relationships with other species of the same genus endemic from other river basins. The results revealed a diploid number of 2n = 56 and the karyotype composed of 18 metacentric, 20 submetacentric, 10 subtelocentric and 8 acrocentric chromosomes (NF = 104). The AgNORs and CMA3 signals were coincident in location occupying the short arm of an acrocentric chromosome pair (25th), in a secondary constriction. The 5S rDNA genes were localized on the short arms of one subtelocentric pair. C-banding revealed terminal blocks on the short arms on many chromosomes as well as terminal positive bands at the both ends of a submetacentric pair. C banding also revealed a large heterochromatic block in the secondary constriction (25th) region that was coincident with the AgNORs sites and CMA3+ bright bands. In spite S. doceanum represent an endemic taxon, in spite their geographic isolation their cytogenetic characteristics show similarities with other species of the genus.(AU)

O presente trabalho apresenta a primeira descrição citogenética de Steindachneridion doceanum, grande bagre que se encontra atualmente em alto risco de extinção e listado como ameaçado na lista vermelha do Ministério do Meio Ambiente, também sugere relações cariotípicas com outras espécies do mesmo gênero, endêmicas de outras bacias hidrográficas. Os resultados revelaram um número diplóide de 56 cromossomos e o cariótipo composto por 18 elementos metacêntricos, 20 submetacêntricos, 10 subtelocêntricos e 8 acrocêntricos (NF = 104). As marcações AgNORs e CMA3 foram coincidentes ocupando o braço curto de um par de cromossomos acrocêntricos (par 25), em uma constrição secundária. Os genes 5S rDNA foram detectados nos braços curtos de um par subtelocêntrico. A banda C revelou blocos terminais nos braços curtos em vários cromossomos, bem como blocos terminais nas duas extremidades de um par submetacêntrico. A banda C também evidenciou um grande bloco heterocromático na constrição secundária (par 25) coincidente com os sítios AgNORs e as bandas CMA3 positivas. Apesar de S. doceanum representar um táxon endêmico, suas características citogenéticas mostram semelhanças com outras espécies do gênero das quais se encontra geograficamente isolado.(AU)

Cytogenetic characterization of Hoplias malabaricus (Bloch, 1794) from the Ctalamochita River (Córdoba, Argentina): first evidence for southernmost populations of this species complex and comments on its biogeography.

Hoplias malabaricus (Bloch, 1794), a predatory freshwater fish with a wide distribution throughout South America, represents a species complex with seven well characterized karyomorphs at the cytogenetic level. Although this species has been extensively studied in several Brazilian basins, data are still scarce for hydrographic systems from other South American countries. This study aims to characterize cytogenetically the Hoplias malabaricus populations from the Argentinean Central Region, close to the southernmost distribution of this species complex. A total of 32 specimens from the Ctalamochita River, a tributary of Lower Paraná Basin located in the province of Córdoba, were analyzed using cytogenetic techniques (Giemsa staining, C- and Ag-NOR banding and fluorescent in situ hybridization with 18S rDNA). All the specimens showed diploid number 2n=42, chromosomic formula 22m + 20sm and absence of sexual chromosomes. Thus, the analyzed populations belong to the karyomorph named A. These populations showed a remarkable degree of divergence in their cytogenetic traits such as karyotypic formula, C-banding, NORs and 18S rDNA patterns for Hoplias malabaricus from other populations bearing the same karyomorph in the Middle and Upper Paraná Basin. These findings are consistent with molecular data from a recent study (where specimens collected in the present work were included), which indicate a closer phylogenetic relationship of Hoplias malabaricus populations from the Ctalamochita River with those from the Uruguay basin and the coastal regions of South Brazil than with populations from the Middle and Upper Paraná Basin. Overall, these pieces of evidence highlight the distinctive features of Hoplias malabaricus from the Ctalamochita River, and also reveal a complex history of dispersion of these populations. The present work is the first to provide cytogenetic information and include some phylogeographic aspects of Hoplias malabaricus populations living in close proximity to the southernmost extreme of its distribution area. Therefore, this study expands significantly upon the previously known geographical coverage for karyomorph A and contributes to a better understanding of the karyotypic diversification within this species complex.

First report of B chromosomes in three neotropical thorny catfishes (Siluriformes, Doradidae).

The family Doradidae (Siluriformes) is an important group of fishes endemic to freshwater ecosystems in South America. Some cytogenetic studies have been conducted focused on the group; however, there are no reports on the occurrence of B chromosomes for the family. In this paper the chromosomal characteristics of Platydoras armatulus (Valenciennes, 1840), Pterodoras granulosus (Valenciennes, 1821) and Ossancora punctata (Kner, 1855) were investigated through classical cytogenetics approaches. The conventional staining reveals 2n=58 in Platydoras armatulus and Pterodoras granulosus, however with distinct karyotypic formulae, possibly originated by pericentric inversions. In Ossancora punctata a derivate karyotype was described with 2n=66 and predominance of acrocentric chromosomes. The C banding pattern was resolutive in discriminating the three species, being considered an important cytotaxonomic marker. All species showed B chromosomes totally heterochromatic with non-Mendelian segregation during meiosis and low frequencies in mitotic cells. The probably origin of these additional elements was through fragmentations of chromosomes of the standard complement, which occurred recently and independently in these three species. The diploid number observed in Ossancora punctata is an evidence of centric fusions and up to the moment it is the highest diploid number reported for Doradidae.

Cytogenetic studies in the redtail catfish, Phractocephalus hemioliopterus (Bloch & Schneider, 1801) (Siluriformes, Pimelodidae) a giant fish from Amazon basin.

The objective of this study was to cytogenetically analyze Phractocephalus hemioliopterus comparing the findings with other data to infer relationships among Pimelodidae species. The results revealed a diploid number of 2n = 56 and the karyotype composed of 16 metacentric, 20 submetacentric, 6 subtelocentric and 14 acrocentric chromosomes (FN = 98). The Ag-NORs, 18S rDNA and CMA3 signals were coincident in location occupying the short arm of an acrocentric chromosome pair (23th), in a secondary constriction. The 5S rDNA genes were localized near the centromere on the short arms of one submetacentric chromosome pair. C-bands were localized predominantly in the terminal regions of chromosomes, including the AgNORs and a small metacentric pair with a conspicuous positive band on interstitial region. This chromosome pair could be considered a species-specific cytogenetic marker.

The importance of pericentric inversions in the karyotypic diversification of the species Loricariichthys anus and Loricariichthys platymetopon.

Catfishes of the genus Loricariichthys are widely distributed in the Platina Basin. Considering that the cytogenetic knowledge of Loricariichythys is underestimated, this study assessed Loricariichthys anus and Loricariichthys platymetopon through different chromosome bandings, to define the mechanisms determining the variability in these species. Cytogenetic analyses evidenced a high degree of similarity in relation to the 2n (54 chromosomes), as well as to the distribution of heterochromatin. Despite the apparent conservatism, it was possible to differentiate between these species, especially in relation to the location of the 18S rDNA genes. An interpopulation variation in the karyotype formula was detected only in L. anus, showing the existence of different karyotypes, probably due to the geographical isolation between Laguna dos Patos and Tramandaí River. The maintenance of the 2n=54, along with the different karyotypes observed in L. anus, the differential nucleolus organizer regions position, as well as the sexual chromosome system ZZ/ZW in L. platymetopon, makes the participation of pericentric inversions in the karyotypic evolution of these species evident. These structural rearrangements were important for chromosome evolution of these two species, because they probably promoted the postzygotic barriers to reproduction, significantly contributing to the speciation process between them.

Cytogenetics of the Porthole Shovelnose Catfish, Hemisorubim platyrhynchos (Valenciennes, 1840) (Siluriformes, Pimelodidae), a widespread species in South American rivers.

Hemisorubim platyrhynchos is a medium- to large-sized pimelodid catfish distributed along several river basins of the Neotropical Region, noteworthy for representing an important fishery source. In this work, Hemisorubim platyrhynchos from three isolated populations were cytogenetically analyzed. The karyotype shows a diploid number of 2n=56 chromosomes comprising 22m, 16sm, 10st, 8a (FN=104). NORs detected by AgNO3 were located in the terminal regions of the short arm of a st chromosome pair, as confirmed by CMA3 and FISH using an 18S rDNA probe. C-banding revealed a small amount of heterochromatin in chromosomes, including the NORs, and one biarmed pair that showed conspicuous positive bands on both arms. This fact was also evidenced when using other banding techniques, such as RE (AluI), and indicates that this pair constitutes a species-specific cytogenetic marker.

Cytogenetic description of Bunocephalus doriae Boulenger, 1902 (Siluriformes: Aspredinidae) from the Paraná River (Misiones, Argentina)

In this work, Bunocephalus doriae was cytogenetically analyzed. A karyotype with a diploid number of 2n= 50 comprising 6m, 10sm, 6st, and 28a (FN= 72) chromosomes was observed. The occurrence of an asymmetric karyotype with a large number of acrocentric chromosomes distinguishes this species from others the Order Siluriformes. An exclusive character observed is the first pair of subtelocentric as the largest chromosome pair of the complement. NORs detected using AgNO3 were located in the terminal regions, on the short arm of a subtelocentric chromosome pair (pair 11), in a secondary constriction. C-banding revealed heterochromatic centromeric regions on several chromosomes of the complement after C-banding. This is the first cytogenetic description of this species and the first cytogenetic report on a member of the family Aspredinidae.

No presente trabalho, Bunocephalus doriae. foi analisado citogeneticamente. O número diploide encontrado foi 2n= 50 compreendendo 6m, 10sm, 6st, and 28a (NF= 72) cromossomos. A ocorrência de um cariótipo assimétrico com um grande número de cromossomos acrocêntricos distingue esta espécie das demais pertencentes à Ordem Siluriformes e foi observada como característica exclusiva a presença do primeiro par subtelocêntrico, sendo o maior do complemento. As NORs detectadas pelo AgNO3 foram observadas na região terminal do braço curto de um par cromossômico subtelocêntrico (par 11), em uma constrição secundária. A heterocromatina, após o bandamento C, foi visualizada em regiões centroméricas de vários cromossomos do complemento. Esta é a primeira descrição citogenética desta espécie e a primeira descrição de um membro da família Aspredinidae.

Cytogenetic description of Bunocephalus doriae Boulenger, 1902 (Siluriformes: Aspredinidae) from the Paraná River (Misiones, Argentina)

In this work, Bunocephalus doriae was cytogenetically analyzed. A karyotype with a diploid number of 2n= 50 comprising 6m, 10sm, 6st, and 28a (FN= 72) chromosomes was observed. The occurrence of an asymmetric karyotype with a large number of acrocentric chromosomes distinguishes this species from others the Order Siluriformes. An exclusive character observed is the first pair of subtelocentric as the largest chromosome pair of the complement. NORs detected using AgNO3 were located in the terminal regions, on the short arm of a subtelocentric chromosome pair (pair 11), in a secondary constriction. C-banding revealed heterochromatic centromeric regions on several chromosomes of the complement after C-banding. This is the first cytogenetic description of this species and the first cytogenetic report on a member of the family Aspredinidae.

No presente trabalho, Bunocephalus doriae. foi analisado citogeneticamente. O número diploide encontrado foi 2n= 50 compreendendo 6m, 10sm, 6st, and 28a (NF= 72) cromossomos. A ocorrência de um cariótipo assimétrico com um grande número de cromossomos acrocêntricos distingue esta espécie das demais pertencentes à Ordem Siluriformes e foi observada como característica exclusiva a presença do primeiro par subtelocêntrico, sendo o maior do complemento. As NORs detectadas pelo AgNO3 foram observadas na região terminal do braço curto de um par cromossômico subtelocêntrico (par 11), em uma constrição secundária. A heterocromatina, após o bandamento C, foi visualizada em regiões centroméricas de vários cromossomos do complemento. Esta é a primeira descrição citogenética desta espécie e a primeira descrição de um membro da família Aspredinidae.

Chromosomal analyses in Megalonema platanum (Siluriformes: Pimelodidae), an endangered species from South American rivers

Este estudo apresenta dados cromossômicos de Megalonema platanum do rio Tibagi, Paraná, Brasil e do rio Paraná, Argentina. O número diploide foi igual 54 com composição cariotípica de 24m+16sm+2st+12a em ambas populações. Os sítios AgNORs foram detectados na posição terminal de um par submetacêntrico das duas populações analisadas, coincidindo com constrição secundária no braço curto do par 15. CMA3 e FISH com sonda de DNAr 18S exibiram sinais fluorescentes que correspondem aos sítios AgNORs e à constrição secundária. A presença de um pequeno cromossomo supranumerário acrocêntrico foi observado em M. platanum do rio Tibagi, com heterocromatina centromérica. Outros blocos heterocromáticos foram evidenciados na posição terminal de alguns cromossomos e um par cromossômico submetacêntrico grande, provavelmente o primeiro par, mostrou heterocromatina intersticial. Na população do rio Paraná foram observados ainda blocos heterocromáticos em ambas regiões terminais em alguns cromossomos. Este trabalho mostra pela primeira vez dados citogenéticos de M. platanum, que é uma espécie muito rara na bacia do rio Paraná e pode estar ameaçada de extinção.

This study presents chromosomal data of Megalonema platanum from rio Tibagi, Paraná, Brazil and from rio Paraná, Argentina. The diploid number was equal 54 with karyotype composition of 24m+16sm+2st+12a in both populations. The AgNOR siteswere detected in the terminal position of a submetacentric pair of the two analyzed populations, coinciding with secondary constrictions on the short arm of pair 15. CMA3 and FISH with 18S rDNA probe displayed fluorescent signals that correspond to the AgNOR sites and secondary constriction. The presence of a small acrocentric supernumerary chromosome can be observed in M. platanum from rio Tibagi, with centromeric heterochromatin. Others heterochromatic blocks were evidenced in the terminal position of some chromosome and one metacentric large chromosome pair, probably the first pair, showed an interstitial heterochromatin. In the population of the rio Paraná were still observed heterochromatic blocks in both ends in some chromosomes. This work brings for the first time cytogenetic date of M. platanum, which is a very rare species in the rio Paraná basin and may be endangered.

Analyses of the structure of NORs in two species of South American Sorubiminae fishes (Siluriformes) by means of several cytogenetic techniques.

Several cytogenetic techniques (AgNOR, C- G- and RE bandings, DAPI, CMA3 and FISH) were applied in order to analyze the structure and variability of NORs in the fish species Steindachneridion melanodermatum and S. scripta. Ag-NORs were observed on the short arm of the first acrocentric chromosome pair, coincidentally with a strong C-positive band on a large secondary constriction. In addition, NORs showed bright fluorescent signals when stained with CMA3 and treated for FISH with rDNA 18S. However, they showed negative coloration after G- and restriction enzyme banding and DAPI staining. The results evidence a substantial size polymorphism in these regions. The NOR bearing chromosomes in both species may be considered homologues because they maintain conserved characteristics, such as being interspersed with a GC-rich heterochromatin and possessing target sequences for AluI, BamHI and EcoRI.

Chromosomal studies of five species of the marine fishes from the Paranaguá Bay and the karyotypic diversity in the marine teleostei of the Brazilian coast

In this study, five species of marine fishes from the Paranaguá Bay in the Brazilian coast were evaluated. Eucinostomus argenteus and Diapterus rhombeus (Gerreidae) presented 48 chromosomes, all of which more acrocentric (FN = 48); Strongylura timucu and S. marina (Belonidae) also presented 48 chromosomes, but with a higher karyotypic complexity than the Gerreidae, 10M+2SM+36A (FN = 60) and 4M+44A (FN = 52), respectively. The fifth species, Mugil curema (Mugilidae), different than the others, presented only 28 chromosomes 20M+4ST+4A (FN = 48). The species presented diversity in the karyotypic macro-structure, which should be relevant for the cytotaxonomy and the evolution of this group of the vertebrate.

Nas últimas décadas tem ocorrido no Brasil um incremento de estudos cariotípicos em peixes marinhos. Atualmente são conhecidos os cariótipos de 118 espécies, distribuídas em 43 famílias e 80 gêneros. Foram estudadas cinco espécies de peixes marinhos do complexo estuarino da Baía de Paranaguá na costa brasileira. Eucinostomus argenteus e Diapterus rhombeus (Gerreidae), apresentaram 48 cromossomos todos acrocêntricos (NF = 48); Strongylura timucu e S. marina (Belonidae) apresentaram 48 cromossomos, porém com complexidade cariotípica maior do que apresentada pelos gerreídeos, 10M+2SM+36A (NF = 60) e 4M+44A (NF = 52), respectivamente. A quinta espécie, Mugil curema (Mugilidae), ao contrário das outras quatro espécies aqui analisadas, apresentou apenas 28 cromossomos 20M+4ST+4A (NF = 48). Apesar da tendência em se verificar um cariótipo constituído por 48 cromossomos em teleósteos marinhos, as espécies aqui analisadas apresentam uma diversidade para a macroestrutura cariotípica a ser considerada para a citotaxonomia e evolução desse grupo de vertebrados.

Population analysis of a chromosome polymorphism in Astyanax (Teleostei, Characiformes) species endemic to the Iguaçu River

Chromosomal analyses were performed in the fish Astyanax sp.D collected from three different points: two streams from the right bank and one from the left bank of the Upper Iguaçu River, Paraná State, Brazil. The individuals from all localities possess 2n = 50 chromosomes and a FN = 84 (4m+24sm+6st+16a). The C-banding pattern was similar in all populations. However, within each population, an interindividual variation concerning the number and localization of heterochromatic bands was observed. Some of these variations were quantified in each population, and the results indicate that the samples were not different when studying the variable frequencies. Considering that Astyanax sp.D is typical in the headwaters of the Iguaçu River, these results were not expected. The data indicate that gene flow is occurring and that the Iguaçu River is not an ecological barrier among the Astyanax sp. D populations.

Cytotaxonomy, heterochromatic polymorphism and natural triploidy of a species of Astyanax (Pisces, Characidae) endemic to the Iguaçu river basin

Cytogenetic analysis with Astyanax sp. D revealed a karyotype of 2n=50 with 2M+26SM+6ST+16A, besides a triploid specimen showing 2n=75 chromosomes (3M+39SM+9ST+24A). C-banding strongly stained the terminal regions of several SM-ST-A chromossomes. Two pairs of acrocentric chromosomes presented interstitial heterochromatin, this state being polymorphic and occuring due to possible paracentric inversions. The results obtained with the AluI restriction enzyme and A3 chromomycin were similar to the C-banding. Relationships were proposed between Astyanax sp. D and A. scabripinnis, as well as considerations for a possible origin of the triploid specimen (2n=3x=75). When comparing the present results with cytogenetic features of other endemic Astyanax species in the Iguaçu river (A. sp. B and C), a clear differentiation was observed between them, indicating cytogenetics as an important cytotaxonomic tool.

Análises citogenéticas em Astyanax sp. D evidenciaram 2n=50 cromossomos e um cariótipo com 2M+26SM+6ST+16A. O bandamento C destacou as regiões teloméricas de diversos cromossomos SM-ST-A. Dois pares de cromossomos acrocêntricos possuem heterocromatina intersticial, sendo este estado polimórfico decorrente de prováveis inversões paracêntricas. O resultados obtidos com a enzima de restrição AluI e a Cromomicina A3 foram semelhantes aos do bandamento C. São propostas relações de parentesco entre Astyanax sp. D e Astyanax scabripinnis, bem como considerações sobre a possível origem do exemplar triplóide (2n=3x=75). Ao comparar os resultados deste trabalho com outras espécies de Astyanax do Rio Iguaçu, estas espécies se tornam claramente distinguíveis, evidenciando a citogenética como uma importante ferramenta taxonômica.

Karyotypic characterization of Hydromedusa tectifera (Testudines, Pleurodira) from the upper Iguaçu River in the Brazilian state of Paraná

We present the karyotypic characterization of 26 specimens of the side-necked turtle Hydromedusa tectifera collected in the upper Iguaçu River, Paraná state, Brazil. The turtles were cytogenetically analyzed using Giemsa staining and other banding techniques (C, G, Ag-NOR and CMA3) as well as fluorescence in situ hybridization (FISH) with a rDNA 18S probe. All the specimens showed a diploid number of 58 composed of 22 macro and 36 microchromosomes. The Ag-NOR, CMA3 and FISH techniques permitted the identification and characterization of the chromosome pairs bearing nucleolus organizer regions (NORs), while G-banding facilitated a better recognition and pairing of macrochromosomes. These data agree with some information available in the literature and should be very useful for further cytotaxonomic and cytosystematic studies.

First chromosome data on Steindachneridion scripta (Pisces, Siluriformes, Pimelodidae) from Brazilian rivers: Giemsa, CBG, G-, and RE banding.

A cytogenetic study was performed on the large pimelodid species Steindachneridion scripta (Siluriformes, Sorubiminae) from the Paraná River basin (Brazil). Chromosome preparations were obtained avoiding sacrifice of the specimens, by means of lymphocyte culture, and several staining and chromosome banding techniques were applied. The karyotype consisted of 56 chromosomes, 24 metacentrics, 20 submetacentrics, 4 subtelocentrics, and 8 acrocentrics (fundamental number = 104). The first pair of acrocentric chromosomes (pair 25) consistently had a decondensed secondary constriction; the C-banding pattern of some chromosomes allows them to be considered cytogenetic markers (i.e., pairs 1, 3, 4, 6, 7, 9, 13, 23, and 24). G-banding and restriction enzymes provided patterns that helped improve chromosome pairing. This is the first report on a Neotropical pimelodid species of economic interest using several cytogenetic techniques and providing an integral karyotypic characterization.

Mutagenic effects of tributyltin and inorganic lead (Pb II) on the fish H. malabaricus as evaluated using the comet assay and the piscine micronucleus and chromosome aberration tests

Genotoxicity studies on toxic metals and their organic compounds are very important, especially so in the investigation of the effects of these compounds on the aquatic environments where they tend to accumulate. The use of endemic aquatic organisms as biological sentinels has proved useful to environmental monitoring. We assessed the mutagenic potential of tributyltin (TBT) and inorganic lead (PbII) using samples of the fish Hoplias malabaricus (commonly called traíra) using the comet assay and the piscine micronucleus and chromosome aberration tests. Eighteen H. malabaricus were acclimatized in three individual aquariums, each containing six fish, six fish being exposed to 0.3 mg/g of body weight (bw) of TBT, six to 21 mg/g bw of PbII and six being used as controls. Exposure to TBT and PbII was achieved by feeding the fish every five days with Astyanax (a small fish that is part of the normal diet of H. malabaricus) which had been injected with solutions of TBT, PbII or with water (the control group). After two months the H. malabaricus were sacrificed and their peripheral blood collected and subjected to the comet and micronucleus assays, the chromosome aberration assay being conducted using kidney-tissue. Although the comet assay showed now mutagenic effects at the lead concentrations used but encountered results with TBT, the micronucleus and chromosome aberrations assays both indicated that TBT and PbII are potentially mutagenic (p < 0.01), the micronucleus assay showing morphological alterations of the nucleus.

Genetic damage induced by trophic doses of lead in the neotropical fish Hoplias malabaricus (Characiformes, Erythrinidae) as revealed by the comet assay and chromosomal aberrations

The effects of clastogenic or mutagenic agents have rarely been studied in neotropical fish species exposed to contaminated water. In this study, the genetic damage caused by lead in the widely distributed South American fish, Hoplias malabaricus, was assessed using the comet (SCGE) assay and by testing for chromosomal aberrations. Eighteen specimens were acclimatized to laboratory conditions and then chronically exposed to contaminated food by feeding prey (Cyprinus sp.) injected intraperitoneally with doses of inorganic lead adjusted to give a contamination level of 21 mg of Pb2+.g-1 net weight of H. malabaricus. Three fish were sampled for chromosomal analysis after four doses (18 days) and another three after eight doses (41 days) of lead and the results then compared with three untreated controls kept under lead-free conditions. An additional six treated fish and three controls were sampled for the comet assay after 13 doses (64 days). Exposure to lead significantly increased the frequency of chromosomal aberrations and the frequency of tailed cell nuclei, the latter indicating DNA damage. These results show that H. malabaricus is a useful biological model for screening the clastogenic effects of lead and possibly other xenobiotics. The genetic damage seen here illustrates the need to investigate the potential effects of heavy metals on fish species in South America.

Karyotypic characterization and NOR analysis by different banding techniques of Rhamdia quelen (Pisces, Pimelodidae) from the first plateau of the Iguaçu River (Brazil).

A population of R. quelen from the first plateau of the Iguaçu River (Paraná State, Brazil) was analyzed cytogenetically. A diploid set of 58 chromosomes was constituted by 32 M, 16 SM, 6 ST and 4 A (FN=116). In one individual a 2n=59 karyotype was determined due to the presence of one additional metacentric heterochromatic chromosome. NORs, detected by AgNO3 and CMA3 staining as well as fluorescence in situ hybridization with an 18S rDNA probe, are located at a terminal position on the short arms of a ST chromosome pair. C-banding marks the NORs and other weak bands distributed on telomeric regions of some chromosomes. These results give evidence that Rhamdia constitute, in terms of karyotypic macrostructure, a conserved group and support the idea that several synonymous species may be included in this genus.