Contribuição à citogenética de testudines e análise de assimetria flutuante em Eretmochelys imbricata, Cheloniidae

This work has contributed to knowledge of the order Testudines from cytogenetic and morphological point of view. With regard to the aspects proposed cytogenetic characterization of the species Mesoclemmys tuberculata (n = 5), endemic to the Caatinga biomes, through conventional techniques of cytogenetics and molecular levels. This species presented 2n = 58, NF = 64, the first submetacentric pair, the second metacentric and third subtelocentric, and the other microchromosome telocentric. This species showed a nucleolar bearing pair, coincident with the 18S ribosomal rDNA and that proved to be heterochromatic. Small heterochromatic blocks were also found in the centromeres of the largest chromosomes, as well as terminal regions in most other chromosomes of the complement, that were GC +. Telomeric sequences showed variable patterns of signal intensity, with some repeats more intense in microchromosomes and subtly in the larger ones. When compared with other species of the genus, the G-banding patterns showed a marked similarity between them. The first karyotypic description of the species will aid in future studies and the understanding of evolutionary aspects of this family. From the morphological point of view, we carried out studies of fluctuating asymmetry in sea turtle Eretmochelys imbricata, using methods of benchmarking between hatchlings and adults and their implications for natural selection. Data were collected at two different times: first during the spawning female and the second during the outbreak and birth of the nest. The analyzed characteristics consisted of measurements of length and width of front and rear flippers (CANT, LANT, CPOS and LPOS) also collected data on the number of hull plates, side plates (NPL), the surrounding plates (NPCIRC), and plastron; plates power plants (NPP), inframarginais plates (NPIM). With the values of asymmetry we calculated the value of strict heritability for these traits, the calculation was based on only one parent. A nonparametric analysis Mann-Whitneywas performed to compare the groups (females X hatchlings, newborn hatchlings X dead hatchlings). Adult females showed no bilateral fluctuating asymmetry (FA = 0) on the number plates of the hull and plastron, while offspring, living and dead, showed a greater level of variation in these meristic parameters. In the analysis of females x hatchlings we found a significant difference between the levels of asymmetry in hoof plates (p=0.006) an the width of hindlimbs (p=0.001). Levels of FA suggest an accurate indicator as to the viability or maintenance of the individual to the reproductive phase. The coefficient of heritability (h2) of FA , obtained from the regression analysis, showed that both have low and not statistically significant values(p> 0.1). In the case of exclusion of the effective role of genetics in the generation of FA, reproductive strategies based on high number of subsidiaries products, such as those observed in E. imbricata seems to implicate the production of individuals with high level of developmental instability

Microbiology quality in continuous water flow fish ponds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Brycon gouldingi (Teleostei, Characidae): aspects of the embryonic development in a new fish species with aquaculture potential

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Piscinoodinium pillulare (Schäperclaus, 1954) Lom, 1981 (Dinoflagellida) infection in cultivated freshwater fish from the Northeast region of São Paulo State, Brazil: parasitological and pathological aspects

O Centro de Aqüicultura, Unesp, Jaboticabal, SP, Brasil, recebeu peixes para diagnose, os quais apresentavam aglomeração nas bordas dos viveiros e na entrada da água. Dos 194 casos diagnosticados, 53 apresentavam estruturas brancas circulares ou ovais, imóveis, medindo 162 mm de diâmetro, identificadas como o dinoflagelado Piscinoodinium pillulare. em 34 casos, os parasitos estavam presentes nas brânquias, em 2 casos, no corpo e em 9 casos, em ambos. Dos 53 casos observados, 31 eram o híbrido tambacu; 7, o Piaractus mesopotamicus; 6, o Colossoma macropomum; 5, o Leporinus macrocephalus; 3, o Oreochromis niloticus; e 1, o Prochilodus lineatus. Os peixes apresentaram aumento da produção de muco no corpo e nas brânquias e equimoses no pedúnculo caudal e nos opérculos. As brânquias também apresentaram palidez, congestão e petéquias. A histopatologia revelou a presença de grande número de trofontes situados entre as lamelas secundárias, fixados ou não ao epitélio. As lamelas primárias mostraram hemorragias intersticiais, severa hiperplasia do epitélio e das células caliciformes e infiltrado inflamatório. O presente trabalho é o primeiro relato de P. pillulare no Brasil e enfatiza a importância dos dinoflagelados, que causaram significativas perdas econômicas entre 1995 e 1997.

Effects of different sampling intervals on apparent protein and energy digestibility of common feed ingredients by juvenile oscar fish (Astronotus ocellatus)

O presente estudo avaliou a digestibilidade aparente da proteína e da energia de ingredientes (farelo de soja, farinha de peixe, farelo de trigo e milho) por juvenis de apaiari (Astronotus ocellatus) usando dois diferentes intervalos de coleta (30 min. e 12h). Os 160 juvenis de apaiari utilizados (22,37 ± 3,06 g de peso corporal) foram divididos em quatro tanques rede plásticos e cilíndricos, cada um colocado em um tanque de alimentação de 1.000 L. O experimento foi inteiramente casualizado em esquema fatorial 2 x 4 (2 intervalos de coleta de fezes e 4 ingredientes foram) com quatro repetições. Os testes estatísticos não detectaram efeito da interação entre o intervalo de coleta e tipo de ingrediente nos coeficientes de digestibilidade. O intervalo de coleta não afetou a digestibilidade da proteína e da energia. As características físicas das fezes dos juvenis de apaiari aparentemente as tornam menos sensíveis à perda de nutrientes por lixiviação, permitindo intervalos de coleta maiores. A digestibilidade da proteína dos ingredientes avaliados foi semelhante, mostrando que a digestibilidade aparente de ingredientes animais e vegetais por juvenis de apaiari é eficiente. Os coeficientes de digestibilidade da energia foram maiores para a farinha de peixe e o farelo de soja comparado a farelo de trigo e milho. Ingredientes ricos em carboidratos (farelo de trigo e milho) apresentaram os piores coeficientes de digestibilidade da energia e, portanto, não são usados eficientemente pelos juvenis de apaiari.

Structural and functional evidence that a B chromosome in the characid fish Astyanax scabripinnis is an isochromosome

Astyanax scabripinnis possesses a widespread polymorphism for metacentric B chromosomes as large as the largest chromosome pair in the A complement. on the basis of C-banding pattern, it was hypothesized that these B chromosomes are isochromosomes that have arisen by means of centromere misdivision and chromatid nondisjunction. In the present paper we test this hypothesis by analysing (i) the localization of a repetitive DNA sequence on both B chromosome arms, and (ii) synaptonemal complex formation, in order to test the functional homology of both arms. Genomic DNA digested with KpnI and analysed by gel electrophoresis showed fragments in a ladder-like pattern typical of tandemly repetitive DNA. These fragments were cloned and their tandem organization in the genome was confirmed. A 51-bp long consensus sequence, which was AT-rich (59%) and contained a variable region and two imperfect reverse sequences, was obtained. Fluorescence in situ hybridization (FISH) localized this repetitive DNA into noncentromeric constitutive heterochromatin which encompasses the terminal region of some acrocentric chromosomes, the NOR region, and interstitial polymorphic heterochromatin in chromosome 24. Most remarkably, tandem repeats were almost symmetrically placed in the two arms of the B chromosome, with the exception of two additional small clusters proximally located on the slightly longer arm. Synaptonemal complex (SC) analysis showed 26 completely paired SCs in males with 1B. The ring configuration of the B univalent persisting until metaphase I suggests that the two arms formed chiasmata. All these data provided strong support for the hypothesis that the B chromosome is an isochromosome.

Supernumerary chromosomes in the pufferfish Sphoeroides spengleri: first occurrence in marine Teleostean Tetraodontiformes fish

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structural analysis of fertilization in the fish Brycon orbignyanus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Levantamento cariotípico em espécies de peixes marinhos costeiros de fundo arenoso (Osteichthypes, Perciformes)

Cytogenetics analyses in fish are important because they compose a private group among the vertebrates, occupying a central position in the animal evolution. The Perciforms Order, dominant in the marine and freshwater environment, it constitutes a model potentially useful in the genetic evaluation of populations, as well as in the understanding of its evolutionary processes. In spite of this, cytogenetics studies in this great group is scarce, above all for the inhabitants of sandy bottom and pelagics habits. The present work proposed to contribute for the cytogenetic characterization of nine species of fish marine of sandy bottom of the coast of Rio Grande do Norte (Brazil), identifying the evolutionary patterns related to the karyotype in these species and the existence of filogenetics affinities between them and other Perciformes. The animals were collected in the beaches of the Redinha, Ponta Negra and Búzios (Coast of Rio Grande do Norte) and in Saint Peter and Saint Paul Archipelago. Later on they were submitted to the cytogenetics technical that consist of mitotic estimulation, obtaining of mitotics chromosomes, proceeded by techniques of conventional coloration (Giemsa) and chromosomic bands (Ag-RONs and C band). Diploid number and fundamental number equal to 48 were observed in most of the species: Menticirrhus americanus, Ophioscion punctatissimus, Pareques acuminatus (Sciaenidae); Chloroscombrus chrysurus (Carangidae); Echeneis sp. 2 (Echeneidae); Archosargus probatocephalus (Sparidae) and Orthopristis ruber (Haemulidae). Trachinotus goodei (NF=52) (Carangidae) and Echeneis sp. 1 (Echeneidae) (NF=54) presented variation in NF, staying constant a diploid number equal to 48. RONs was situated in pericentromeric position in whole the scianids, and in the species Echeneis sp. 2 (22° pair), O. ruber and A. probatocephalus (1° pair), coinciding with great heterocromatics blocks in M. americanus (1° pair), P. acuminatus (2° pairl) and O. ruber (1° pair). RONs was also located in the telomeric area of the short arm of the 5° and 11° acrocentrics pairs in T. goodei, 4° and 19° pairs of C. chrysurus, 1° pair (sm) of Echeneis sp. 1. The C band detected centromeric blocks in most of the chromosomes of the species of Sciaenidae, Carangidae and Echeneidae, with great blocks in A. probatocephalus (4° pair). Heterocromatic blocks in telomeric areas in submetacentrics of Echeneis sp. 1, and pericentromerics in M. americanus (1° and 8° pairs), O. punctatissimus (1° pair) and P. acuminatus (2° pair) were also observed. It is noticed a marked conservatism cromossomic in the species of the family Scianidae and Haemulidae in what says respect to the number of acrocentrics chromosomes and the location of RONs. Even so it is outstanding the presence of heterocromatinization events during the karyotypic evolution of this family. Already in the families Sparidae and Carangidae, the obtained results reaffirm examples of small variations structural resultants of inversion and translocation Robertsonian, as important mechanisms of diversification karyotipical, as well as a pattern numerical evolutionary conserved, also observed in representatives of Echeneidae of Atlantic in relation to Pacific. The presence of RONs multiple, observed in the species T. goodei and C. chrysurus seems to represent a character derived in the family Carangidae. The results for the species O. ruber and A. probatocephalus suggest the presence of possible geographical or climatic barriers among populations of NE of Brazil in relationship the one of the SE

Padrões cariotípicos em tetraodontiformes (Osteichthypes) :redução genômica e mecanismos de diversificação

The Tetraodontiformes order is composed for about 400 species of fish, distributed in ten families, with circuntropical distribution. The morphologic diversity of each family reflects in, part, the different levels of specialization. This group represents an ancestry after-Perciformes and constitutes the last branch of the diffusion of the Teleósteos, occupying a position of prominence. The phylogenetics relationships of the Tetraodontiformes exist diverse works examining and, in all, these families are recognized as groups brothers, being Diodontidae next to Tetraodontidae and Balistidae next to Monacanthidae. Although it possesss a representative number of species, the works involving of the families Balistidae and Monacanthidae are few exemplary, especially species of oceanic islands. In this work cytogenetic studies in five species had been analyzed Cantherhines macrocerus, Cantherhines pullus (Monacanthidae), Melichthys niger (Balistidae), Sphoeroides testudíneus (Tetraodontidae) and Chilomycterus antennatus (Diodontidae); through conventional coloration, Ag-NORs and C banding. Ahead of the different karyological trends of evolution presented by the Tetraodontiformes, the present work also searched to verify the relation existence enters the total size of the chromosomes with the amount of DNA in these groups of Tetraodontiformes. For such, they had been correlated the total size of the chromosomes of these species, with values of content of available DNA in literature. The cytogenetics analyses for the species C.macrocerusJ C.pullus (Monacanthidae) and M.niger (Balistidae), had disclosed 40 chromosomes, all acrocentrics. All possess only one pair of NORs and pericentromeric heterochromatin. For S.testudíneus the found dyploid number was equal 2n=46, with NF=78 (16m+18sm+8st+4a), while that for C.antennatus it possesss 2n=50, with NF=76 (4m+22st+24a). Both species possess simple NORs and pericentromeric heterochromatin blocks. In M.niger, the presence of positive marking (heterochromatin and NOR) in the secondary constriction in the second chromosomic pair suggesting the occurrence of a rearrangement, possibly a fusing involving these homologous ones, indicating that these events had been important for the establishment of the karyological history of this group. A maintenance of the chromosomic constancy found in the populations of C.macrocerus (Monacanthidae) and S.testudineus (Tetraodontidae) perhaps if must for the aiding of the gene flow through oceanic chains. These data contrast with the differentiated kinds of chromosomes of C.antennatus between the Northeast coast and Southeastern, suggesting that the ecological standards of each species, added to the conditions of the marine environment, can be responsible for the karyological delineation of each species. The found characteristics for the species C.macrocerus, C.pullus, M.niger, S.testudineus and C.antennatus add it the available data for other species of Tetraodontiformes. From the data gotten in the present study, it can be inferred that the DNA content possesss direct relation with the total length of the chromosomes

Effects of mercury intoxication on the response of horizontal cells of the retina of thraira fish (Hoplias malabaricus)

Methyl mercury (MeHg) is highly neurotoxic, affecting visual function in addition to other central nervous system functions. The effect of mercury intoxication on the amplitude of horizontal cell responses to light was studied in the retina of the fish Hoplias malabaricus. Intracellular responses were recorded from horizontal cells of fish previously intoxicated with MeHg by intraperitoneal injection (IP group) or by trophic exposure (T group). Only one retina per fish was used. The doses of MeHg chloride administered to the IP group were 0.01, 0.05, 0.1, 1.0, 2.0, and 6.0 mg/kg. The amplitudes of the horizontal cell responses were lower than control in individuals exposed to 0.01 (N = 4 retinas), 0.05 (N = 2 retinas) and 0.1 mg/kg (N = 1 retina), whereas no responses were recorded in the 1.0, 2.0, and 6.0 mg/kg groups. T group individuals were fed young specimens of Astyanax sp previously injected with MeHg corresponding to 0.75 (N = 1 retina), 0.075 (N = 8 retinas) or 0.0075 (N = 4 retinas) mg/kg fish body weight. After 14 doses, one every 5 days, the amplitude of the horizontal cell response was higher than control in individuals exposed to 0.075 and 0.0075 mg/kg, and lower in individuals exposed to 0.75 mg/kg. We conclude that intoxication with MeHg affects the electrophysiological response of the horizontal cells in the retina, either reducing or increasing its amplitude compared to control, and that these effects are related to the dose and/or to the mode of administration.

Bioaccumulation of chlorinated pesticides and PCBs in the tropical freshwater fish Hoplias malabaricus: Histopathological, physiological, and immunological findings

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Mercury and DDT exposure risk to fish-eating human populations in Amazon

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Spermatozoon ultrastructure and sperm cryopreservation of the Brazilian dry season spawner fish pirapitinga, Brycon nattereri

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Systemic response induced by Scorpaena plumieri fish venom initiates acute lung injury in mice

Scorpaena plumieri venomous fish inflicted severe injuries in humans characterized by systemic effects and cardiovascular abnormalities. Although cardiotoxic and hypotensive effects induced in rats by this venom have been studied, little is known about their effect on bronchial epithelial permeability and airway inflammation in mice. The primary goal of this study was to determine whether the intraplantar or intraperitoneal injection of S. plumieri venom results in systemic response, and whether this event initiates acute lung injure. We found that BALB/c mice developed neutrophilic infiltrates, areas of lung hemorrhage and alveolar macrophage activation within 24 h after injection with S. plumieri venom. These histopathological changes were associated with an early increase in BAL fluid protein and early induction of cytokines, chemokines and matrix metaloproteinases, followed by a later increase in BAL fluid neutrophils. These findings provide clear evidence that the injection of S. plumieri venom in footpad or peritoneal cavity of mice results in venom deposition in the airway and initiates a sustained inflammatory response in the lungs. (C) 2007 Elsevier Ltd. All rights reserved.