The ultrastructural aspects of vitellogenesis or oocyte secondary growth in Serrasalmus spilopleura (Teleostei, Characiformes, Serrasalminae)

Oocyte secondary growth in S. spiloleura corresponds to the period in which different vesicular structures are formed, including the cortical alveoli and the yolk granules. The oocytes with cortical alveolus formation show vesicular structures with filamentous content in the cortical cytoplasmic region, which are the cortical alveolus precursors. In these oocytes, electron-dense vesicles of heterogenous content are dispersed in the inner cytoplasmic region and their nuclei are irregular, showing many nucleoli of different sizes. The oocytes in vitellogenesis are filled with many vesicles. The cortical alveolus precursors are in the peripheral region, and electron-dense granules are seen near to the nucleus. These fuse and form yolk granules. The oocytes in vitellogenesis show a very irregular nucleus that has nucleoli of different sizes. In the oocytes in final vitellogenesis, the yolk granules are scattered throughout the cytoplasm, displacing the cortical alveoli toward cell periphery. The nucleus is similar to the other stages.

Morfologia da micrópila e da superfície dos ovócitos de piracanjuba, Brycon orbignyanus (Osteichthyes, Characidae), sob microscopia eletrônica de varredura

This paper studied the morphology of oocytes of Brycon orbignyanus, (Osteichthyes, Characidae), through observation in Scanning Electron Microscopy (SEM). Fragments of the ovaries from adult females were collected. During the reproductive period, a hormonal induction in females was carried out to collect the oocytes after extrusion. The samples were fixed and processed for observation in SEM. Results showed that the oocytes of B. orbignyanus had a follicular epithelium formed by a single layer of cells with compressed shape, covering the whole radiatta zone that showed a smooth and regular surface with innumerable pores. The micropyle had a funnel-shaped, containing several furrows. The oocyte surface around the micropyle presented pores closer of each other than the other surface areas of radiatta zone.

Spatial distribution of characidiin fishes (Teleostei, Crenuchidae) in the Ribeirão Grande system, a tributary of Paraíba do Sul river basin, Brazil

This study analyses the spatial distribution of characidiin fishes Characidium lauroi and Characidium alipioi (Crenuchidae) in a forest stream system located in southeastern Brazil. Fish were sampled from Jury 2001 to April 2002. Collections were made with an electro-fishing device in five stream reaches of the Ribeirão Grande system. Conductivity, pH, water temperature and dissolved oxygen were measured at each site. The species have different distributions in Ribeirão Grande system. Characidium lauroi is abundant in montane-piedmont zones and Characidium alipioi occurs mainly in piedmont-plain zones. Streams' different features contribute to these species' distribution in the system.

Cytogenetic characterization of the silverside fish Odontesthes regia (Humboldt, 1833) (Teleostei: Atheriniformes: Atherinopsidae) from Iquique, Chile

This paper describes the karyotype of Odontesthes regia by means of Giemsa staining, C-banding, to reveal the distribution of the constitutive heterochromatin, and by Ag-staining and fluorescent in situ hybridization (FISH), to locate ribosomal genes (rDNA). The chromosome diploid modal count in the species was 2n = 48. The karyotype is composed of one submetacentric pair (pair 1), 16 subtelocentric pairs (pairs 2 to 17), and 7 acrocentric pairs (pairs 18 to 24). With the exception of pair 1 it was not possible to classify the homologous chromosomes accurately because differences in chromosome size were too slight between adjacent pairs. The distribution of C-banded heterochromatin allowed for a more accurate matching of the majority of chromosomes of the subtelocentric series. Silver staining of metaphase spreads allowed for the identification of Nucleolus Organizer Regions (Ag-NOR) on pair 1. FISH experiments showed that 18S rDNA sequences were located, as expected, in the same chromosome pair identified as the Ag-NOR-bearing one.

α-Lipoic acid-enrichment partially reverses tissue ascorbic acid depletion in pacu (Piaractus mesopotamicus) fed vitamin C-devoid diets

Effects of dietary α-lipoic acid (LA) and ascorbic acid (AA) on the growth, tissue vitamin C and tocopherol (vitamin E) levels, and malondialdehyde levels were examined in the tropical fish pacu, Piaractus mesopotamicus. Pacu juveniles were fed one of four casein-gelatin-based diets for 8 weeks: with 0.05% AA and 0.1% LA (+AA+LA), with AA and without LA (+AA-LA), without AA and with LA (-AA+LA), and without AA and LA (-AA-LA). When the fish received quantities of feed equal to 1.9-2.5% of its body weight, growth was not influenced, regardless of the presence of AA or LA throughout most of the experimental period. Growth was, however, slightly but significantly lower at week 8 in the AA-deficient/LA-supplemented group. An AA-deficient diet caused a highly significant reduction in both total AA and dehydroascorbic acid content in the liver and gill tissues. This reduction of tissue AA concentrations was reversed in a significant manner by LA (antioxidant-sparing effect). The 8-week-long vitamin C deprivation was sufficient to initiate the reduction in tissue ascorbic acid; however, total ascorbate in the liver of fish in the (-)AA/(+)LA group was 127.7±54.3 nmol g-1 tissue, whereas it was 28.6±26.3 nmol g-1 in the (-)AA/(-)LA group, a 4.4-fold difference. This mitigating effect of the addition of the endogenous antioxidant LA to the diet indicates that LA exerts a vitamin C-sparing effect in teleost fish that by far exceeds the phenomena demonstrated in non-scurvy-prone mammals. There was no difference among the different diet groups for vitamin E and malondialdehyde levels in the liver. These results suggest that LA is a potent substance for the prevention of AA deficiency in cultured fishes. The optimal dietary level of LA needs to be determined in the light of the slight reduction in body weight gain after 8 weeks of feeding in the absence of AA. © Springer Science+Business Media, Inc. 2006.

Respiratory chemoreceptor function in vertebrates - Comparative and evolutionary aspects

The sensing of blood gas tensions and/or pH is an evolutionarily conserved, homeostatic mechanism, observable in almost all species studied from invertebrates to man. In vertebrates, a shift from the peripheral O2-oriented sensing in fish, to the central CO2/pH sensing in most tetrapods reflects the specific behavioral requirements of these two groups whereby, in teleost fish, a highly O2-oriented control of breathing matches the ever-changing and low oxygen levels in water, whilst the transition to air-breathing increased the importance of acid-base regulation and O2-related drive, although retained, became relatively less important. The South American lungfish and tetrapods are probably sister groups, a conclusion backed up by many similar features of respiratory control. For example, the relative roles of peripheral and central chemoreceptors are present both in the lungfish and in land vertebrates. In both groups, the central CO2/pH receptors dominate the ventilatory response to hypercarbia (60-80), while the peripheral CO2/pH receptors account for 20-30. Some basic components of respiratory control have changed little during evolution. This review presents studies that reflect the current trends in the field of chemoreceptor function, and several laboratories are involved. An exhaustive review on the previous literature, however, is beyond the intended scope of the article. Rather, we present examples of current trends in respiratory function in vertebrates, ranging from fish to humans, and focus on both O2 sensing and CO2 sensing. As well, we consider the impact of chronic levels of hypoxia - a physiological condition in fish and in land vertebrates resident at high elevations or suffering from one of the many cardiorespiratory disease states that predispose an animal to impaired ventilation or cardiac output. This provides a basis for a comparative physiology that is informative about the evolution of respiratory functions in vertebrates and about human disease. Currently, most detail is known for mammals, for which molecular biology and respiratory physiology have combined in the discovery of the mechanisms underlying the responses of respiratory chemoreceptors. Our review includes new data on nonmammalian vertebrates, which stresses that some chemoreceptor sites are of ancient origin.

Cortisol response to acute stress in jundiá Rhamdia quelen acutely exposed to sub-lethal concentrations of agrichemicals

Exposure to agrichemicals can have deleterious effects on fish, such as disruption of the hypothalamus-pituitary-inter-renal axis (HPI) that could impair the ability of fish to respond to stressors. In this study, fingerlings of the teleost jundiá (Rhamdia quelen) were used to investigate the effects of the commonly used agrichemicals on the fish response to stress. Five common agrichemicals were tested: the fungicide - tebuconazole, the insecticide - methyl-parathion, and the herbicides - atrazine, atrazine + simazine, and glyphosate. Control fishes were not exposed to agrichemicals and standard stressors. In treatments 2-4, the fishes were exposed to sub-lethal concentrations (16.6%, 33.3%, and 50% of the LC50) of each agrichemical for 96 h, and at the end of this period, were subjected to an acute stress-handling stimulus by chasing them with a pen net. In treatments 5-7 (16.6%, 33.3%, and 50% of the LC50), the fishes were exposed to the same concentrations of the agrichemicals without stress stimulus. Treatment 8 consisted of jundiás not exposed to agrichemicals, but was subjected to an acute stress-handling stimulus. Jundiás exposed to methyl-parathion, atrazine + simazine, and glyphosate presented a decreased capacity in exhibiting an adequate response to cope with stress and in maintaining the homeostasis, with cortisol level lower than that in the control fish (P < 0.01). In conclusion, the results of this study clearly demonstrate that the acute exposure to sub-lethal concentrations of methyl-parathion, atrazine + simazine, and glyphosate exert a deleterious effect on the cortisol response to an additional acute stressor in the jundiá fingerlings. © 2008 Elsevier Inc. All rights reserved.

An overview of functional and stereological evaluation of spermatogenesis and germ cell transplantation in fish

Although there are almost thirty-thousand species of fish living in a great variety of habitats and utilizing vast reproductive strategies, our knowledge of morphofunctional and quantitative aspects of testis structure and spermatogenesis is still incipient for this group of vertebrates. In this review, we discuss aspects that are important to better understanding of testis structure and function, and of the development of germ cells (GC) during spermatogenesis. To achieve this, we have recently completed a number of studies presenting morphometric and functional data related to the numbers of GC and Sertoli cells (SC) per each type of spermatogenic cyst, the number of spermatogonial generations, the SC efficiency, and the magnitude of GC loss that normally occurs during spermatogenesis. We also investigated SC proliferation and the relationship of this important event to early spermatogenic cysts. The available data strongly suggest that SC proliferation in sexually mature tilapia is the primary factor responsible for the increase in testis size and for determination of the magnitude of sperm production. The influence of temperature on the duration of spermatogenesis in tilapia was also evaluated and we have used this knowledge to deplete endogenous spermatogenesis in this teleost, in order to develop an experimental system for GC transplantation. This exciting technique results in new possibilities for investigation of spermatogenesis and spermatogonial stem cell biology, creating also an entirely new and promising scenario in biotechnology - transgenic animal production and the preservation of the genetic stocks of valuable animals or endangered species. © Springer Science+Business Media B.V. 2008.

Descrição cariotípica de possíveis novas espécies do complexo Hypostomus ancistroides e outros Hypostominae

Cytogenetic analyses were performed in four species of the Hypostominae subfamily, three from Hypostomus (Hypostomini) genus and Rhinelepis aspera (Rhinelepini). Three populations of Hypostomus ancistroides were analyzed, which had 2n=68 chromosomes, but presented different karyotype formulas. Hypostomus regani and H. strigaticeps, both from Ivaí river, showed 2n=72 chromosomes with two distinct cytotypes. In turn, R. aspera of the upper Paraná river basin presented 2n=54 chromosome. Multiple Nucleolar Organizer Regions (NORs) have been evidenced by silver nitrate staining in species of Hypostomus and single NOR in R. aspera. The observed variation in the chromosome number and the marked variability in karyotype formulas and NORs reveal a certain amount of karyotype variation in the genus Hypostomus suggesting the probable existence of cryptic species with independent chromosome traits. Therefore, our data can be of great value in discriminating species and understanding their chromosomal evolution.

Ocorrência de Braga cigarra (Cymothoidae) parasitando Galeocharax knerii (Characidae) de afluentes da represa de Jurumirim, Brasil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Acute aerocystitis in Piaractus mesopotamicus: Participation of eicosanoids and pro-inflammatory cytokines

A total of 360 pacus (Piaractus mesopotamicus) were used to study vascular permeability (VP) and inflammatory cell component (CC) in induced aerocystitis in P. mesopotamicus through inoculation of inactivated Aeromonas hydrophila, and the effect of steroidal and nonsteroidal anti-inflammatory drugs. It was observed that after inoculation of A. hydrophila, the maximum VP occurred 180 min post-stimulus (MPS). Pretreatment with anti-inflammatory drugs inhibited VP, and the inhibitory effect of dexamethasone was seen earlier than the effects caused by meloxicam and indomethacin. Inoculation of the bacterium caused a gradual increase in the accumulation of cells, which reached a maximum 24 h post-stimulus (HPS). Pretreatment with dexamethasone, indomethacin and meloxicam reduced the accumulation of lymphocytes, thrombocytes, granulocytes and macrophages. There was no significant difference between the different doses of the drugs tested. The results suggest that eicosanoids and pro-inflammatory cytokines participate in chemical mediation in acute inflammation in pacus. © 2013 Elsevier Ltd.

The discovery of Foxl2 paralogs in chondrichthyan, coelacanth and tetrapod genomes reveals an ancient duplication in vertebrates

The Foxl2 (forkhead box L2) gene is an important member of the forkhead domain family, primarily responsible for the development of ovaries during female sex differentiation. The evolutionary studies conducted previously considered the presence of paralog Foxl2 copies only in teleosts. However, to search for possible paralog copies in other groups of vertebrates and ensure that all predicted copies were homolog to the Foxl2 gene, a broad evolutionary analysis was performed, based on the forkhead domain family. A total of 2464 sequences for the forkhead domain were recovered, and subsequently, 64 representative sequences for Foxl2 were used in the evolutionary analysis of this gene. The most important contribution of this study was the discovery of a new subgroup of Foxl2 copies (ortholog to Foxl2B) present in the chondrichthyan Callorhinchus milii, in the coelacanth Latimeria chalumnae, in the avian Taeniopygia guttata and in the marsupial Monodelphis domestica. This new scenario indicates a gene duplication event in an ancestor of gnathostomes. Furthermore, based on the analysis of the syntenic regions of both Foxl2 copies, the duplication event was not exclusive to Foxl2. Moreover, the duplicated copy distribution was shown to be complex across vertebrates, especially in tetrapods, and the results strongly support a loss of this copy in eutherian species. Finally, the scenario observed in this study suggests an update for Foxl2 gene nomenclature, extending the actual suggested teleost naming of Foxl2A and Foxl2B to all vertebrate sequences and contributing to the establishment of a new evolutionary context for the Foxl2 gene. © 2013 Macmillan Publishers Limited All rights reserved.

An assessment approach for application of spermatic data in phylogenetic analyses: Within the genus Moenkhausia Eigenmann, 1903 (Characiformes: Characidae)

Spermiogenesis and sperm ultrastructure from 21 species of Moenkhausia and others related genera are described. To evaluate the phylogenetic signals, 18 unordered characters were utilized in implied weighting analysis through the program TNT 1.1. Four variations of spermiogenesis were found. In the earliest spermatids, the nucleus can be positioned lateral, eccentric, strongly eccentric or nearly medial in relation to the distal centriole. The nuclear rotation can be present or absent. These spermiogenesis processes are related or intermediate to Type I and Type III. Taking into account the degrees of nuclear rotation during the spermiogenesis and other characteristics, distinct forms of spermatozoa are observed among the species analyzed. The phylogenetic analysis yielded a single most parsimonious tree with fit value 2.70000 and the topology obtained founds Moenkhausia as non-monophyletic. However, some hypothesis of relationships previously proposed viz the clade 20, which contains the type species Moenkhausia xinguensis, is recovered herein. This clade is supported by five synapomorphies, and it allows the supposition that these species constitute a monophyletic group. The whole topology is presented and discussed. © 2012 The Authors. Acta Zoologica © 2012 The Royal Swedish Academy of Sciences.

Comparative analysis of sex chromosomes in Leporinus species (Teleostei, Characiformes) using chromosome painting

Background: The Leporinus genus, belonging to the Anostomidae family, is an interesting model for studies of sex chromosome evolution in fish, particularly because of the presence of heteromorphic sex chromosomes only in some species of the genus. In this study we used W chromosome-derived probes in a series of cross species chromosome painting experiments to try to understand events of sex chromosome evolution in this family.Results: W chromosome painting probes from Leporinus elongatus, L. macrocephalus and L. obtusidens were hybridized to each others chromosomes. The results showed signals along their W chromosomes and the use of L. elongatus W probe against L. macrocephalus and L. obtusidens also showed signals over the Z chromosome. No signals were observed when the later aforementioned probe was used in hybridization procedures against other four Anostomidae species without sex chromosomes.Conclusions: Our results demonstrate a common origin of sex chromosomes in L. elongatus, L. macrocephalus and L. obtusidens but suggest that the L. elongatus chromosome system is at a different evolutionary stage. The absence of signals in the species without differentiated sex chromosomes does not exclude the possibility of cryptic sex chromosomes, but they must contain other Leporinus W sequences than those described here. © 2013 Parise-Maltempi et al.; licensee BioMed Central Ltd.

Influência de uma piscicultura em tanquesrede na dieta e atributos biológicos de Galeocharax knerii na represa de Chavantes, Brasil

The aim of the present study was to evaluate the diet and biological attributes of the population of Galeocharax knerii residing near net cage fish farming activities in the Chavantes reservoir (Paranapanema River, Brazil) to check their possible impacts. Samples were collected from two populations: one near the net cages (NC) and one from an area not influenced by these cages denominated the reference site (RS). Monthly sampling was carried out from March 2008 to February 2009. Fish were caught with a standardized effort using gill nets deployed for 14 hours. The alimentary index (AI) and degree of repletion (RD) were calculated to determine diet composition. Analyses of the sex ratio and the gonadosomatic index (GSI) were also performed. The calculations of AI revealed that fish wastes constituted the most frequent food item in the diet in both study areas (NC = 70.43; RS = 87.55), followed by the consumption of Apareiodon affinis (AI = 29.56), which was abundant near the NC, and prawn at the reference site (AI = 12.28). The sex ratio differed from 1:1 and mature individuals were only found in the population near the NC. The findings demonstrate that G. knerii indirectly benefits from the input of organic matter, using small fish as its main food resource. We conclude that the activities of fish farming influence diet and biological attributes of the species G. knerii, evidenced mainly by higher feeding activity, numerical abundance and biomass in the area of the cages.