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This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.(AU)

Este estudo teve como objetivo desenvolver um protocolo para a criopreservação do sêmen de Pseudoplatystoma corruscans. Para tal, machos maduros foram induzidos hormonalmente com uma dose única de extrato de hipófise de carpa (5 mg/kg de peso vivo). O sêmen foi coletado e avaliado. Sendo testados para compor os diluentes, dois crioprotetores: dimetil acetamida (DMA) e dimetil sulfóxido (Me2SO), em duas concentrações (8% e 10%), + 5,0% glicose + 10% gema de ovo. O sêmen foi diluído na proporção 1: 4 (sêmen: extensor), embalado em palhetas de 0,5 mL e congelado em container dryshipper em vapores de nitrogênio líquido. Após o descongelamento, foram avaliados os aspectos cinéticos espermáticos, a morfologia espermática e a integridade do DNA dos espermatozoides criopreservados. Os machos de P. corruscans produziram sêmen com motilidade espermática > 80%. Todos os tratamentos proporcionaram após o descongelamento sêmen com motilidade espermática total > 40%, sem diferença significativa (P < 0,05) entre eles, como também entre os demais parâmetros cinéticos espermáticos avaliados. Os tratamentos com DMA proporcionaram uma menor fragmentação do DNA dos gametas. Malformações espermáticas foram identificadas, tanto no sêmen fresco, como no criopreservado, sendo identificado um aumento discreto dessas malformações nos espermatozoides das amostras de sêmen descongeladas de P. corruscans.(AU)

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In 2014, an atypical drought in Southeast Brazil drastically reduced the water level in several reservoirs. We investigated the effects of this drought and the subsequent flood period on the attributes of ichthyofauna in an aquaculture and in a control area. Fish were collected bimonthly between 2014 and 2015 (drought) and 2016 (wet), using gill nets in the two sample areas in the Ilha Solteira reservoir, Upper Paraná River basin, Brazil. We compared ichthyofauna attributes between the drought and wet seasons in each area and between areas within each season. In the aquaculture area, the assemblages showed similar characteristics between the seasons. By contrast, the control area varied between seasons, with greater species richness, Shannon diversity, species evenness, and less ß diversity in the wet season. Comparisons between areas in each season showed higher abundance in the fish farm within the drought season. Changes in structure and composition in the control area are possibly associated with new areas and resources made available by the flooding of marginal areas during the wet season. We inferred that the effect of the flood on the aquaculture community was attenuated by the continuous habitat structure such as shelters and food provided by the enterprise.

Em 2014 um evento de seca atípica no sudeste brasileiro diminuiu drasticamente o nível da água em diversos reservatórios. Nós investigamos os efeitos dessa seca, e seu subsequente período de cheia sobre atributos da ictiofauna em uma área aquícola e uma área controle. Os peixes foram coletados bimestralmente entre 2014 a 2015 (seca) e 2016 (cheia), usando redes de espera nas duas áreas amostrais no reservatório de Ilha Solteira, bacia do alto rio Paraná, Brasil. Comparamos atributos da ictiofauna entre os períodos de seca e cheia em cada área e entre as áreas dentro de cada período. Na área aquícola, verificou-se similaridade das assembleias entre os períodos. Em contraste, a área controle apresentou variação entre os períodos, com maior riqueza, diversidade Shannon, equitabilidade e menor diversidade ß no período de cheia. Comparações entre áreas em cada período, mostraram maior abundância na piscicultura no período de seca. As mudanças na estrutura e composição na área controle possivelmente está associada as novas áreas e recursos disponibilizados pela inundação de áreas marginais no período de cheia. Infere-se que o efeito da cheia na comunidade da área aquícola foi atenuado pela continua estrutura de habitat, como abrigos e ração fornecidos pelo empreendimento.

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Individuals of the same species may present different reproductive tactics depending on the environment in which they develop and mature. The present study aimed to define the gonadal development phases of males and females of Astyanax rivularis and to carry out a comparative analysis of the reproductive development of specimens captured in two isolated environments of the São Francisco River basin in Serra da Canastra, Brazil (Point 1: low vegetation and river showing calm and crystalline waters with small well formations; Point 2: current waters, and well-established areas of arboreal vegetation). Thus, the gonads of A. rivularis specimens were collected, fixed and processed with techniques for light microscopy. Five maturation phases of the females' reproductive cycle were established: immature, developing, spawning capable, regressing and regenerating. Three maturation phases of the males' reproductive cycle were observed: spawning capable, regressing, and regenerating. There are differences in the phases of gonadal development of A. rivularis between the two sampling points so that, possibly, animals upstream of the waterfall demonstrate a delay in the reproductive cycle in relation to animals downstream.

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We aimed to develop a simplified protocol for transplantation of Brycon orbignyanus spermatogonial stem cells (SSCs) into Astyanax altiparanae testes. Brycon orbignyanus testes were enzymatically digested and SSC purified by a discontinuous density gradient. Endogenous spermatogenesis was suppressed in A. altiparanae using busulfan or by incubation at 35 °C water, and SSCs from B. orbignyanus labeled with PKH26 were injected into their testes via the urogenital papilla. Twenty-two hours post-transplantation, labeled spermatogonia were observed in A. altiparanae tubular lumen. After 7 days, spermatogonia proliferated in the epithelium, and 21 days post-transplantation, sperm was observed in the lumen. Of surviving host fish, nearly 67% of those treated with busulfan and 85% of those held in warm water showed labeled cells in host germinal epithelium. The present study standardized, by a simple and accessible method, germ cell transplantation between sexually mature Characiformes fish species. This is the first report of xenogenic SSC transplantation in this fish order.

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In view of the established climate change scenario and the consequent changes in global temperature, it is essential to study its effects on animal spermatogenesis. Therefore, the aim of this study was to verify the duration of spermatogenesis at different temperatures. For this purpose, 96 male and adult specimens of Astyanax altiparanae were kept in a closed circulation system with water temperature stabilized at 27 °C and 32 °C. Subsequently, the specimens received pulses of BrdU (bromodeoxyuridine) at a concentration of 100 mg/kg/day for 2 consecutive days, and the samples were collected daily for a period of 15 days. Their testes were removed, fixed, processed in historesin, and sectioned in 3 µm, submitted to hematoxylin/eosin staining and to bromodeoxyuridine immunodetection. Partial results of the optimum temperature experiments allowed the classification of A. altiparanae spermatogenic cells in Aund, Adiff, and type B spermatogonia, spermatocytes, spermatids, and spermatozoa. The duration of spermatogenesis was determined as approximately 6 days for animals at a temperature of 27 °C and 1 day for animals at 32 °C. The elevated temperature was also responsible for increasing cell proliferation, resulting in an increase in the number of spermatocytes, spermatids, spermatozoa, and cell death (cell pyknotic). The duration of spermatogenesis in A. altiparanae was directly affected by the elevated water temperature, causing a reduction in the estimated time of spermatogenesis.

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This study reports the occurrence of metacercariae of Austrodiplostomum compactum in Pimelodus platicirris from a Neotropical reservoir in the Grande River, SP, Brazil. A total of 164 fish were collected, of which 12.80% were infected with metacercariae in the eyes. The mean intensity of infection and mean abundance were 1.52±0.14 (1‒3) and 0.23±0.05 (0‒3), respectively. The presence of this parasite with a high intensity of infection can cause exophthalmos, retinal displacement, opacity of the lens, blindness or even death. This is the first record of ocular metacercariae for P. platicirris, thus increasing the number of hosts for A. compactum.

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This study aimed to determine the semen characteristics of Astyanax lacustris after hormonal induction and to evaluate the sensitivity of the species sperm to cryoprotective solutions based on the cryoprotectants dimethyl sulfoxide and methyl glycol. Volume, color, sperm concentration, total motility and aspects of sperm movement were analyzed using "Integrated Semen Analysis System". Three different extenders were tested: A) glucose 5%+egg yolk 10%, B) BTS®5% and C) glucose 5% and two permeable cryoprotectants: dimethyl sulfoxide (Me2SO) and methyl glycol (MTG). Fresh A. lacustris semen presented total motility of 76.6±11.2%, motility duration of 33.0±2.2s, sperm concentration of 7.22±3.2×109sptz/mL and seminal osmolality of 219±0.03mOsm/kg-1. The toxicity test showed the highest total motility values at the MTG15%+A, Me2SO15%+B and Me2SO10%+C dilutions, and the Me2SO10%+C and Me2SO15%+C dilutions presented the highest values for curvilinear velocity, linear velocity and average velocity. The tested protocol was not effective at maintaining the viability of A. lacustris semen after freezing because no motility was observed in any of the dilutions. However, the Comet Assay demonstrated that cryoprotectant solutions were effective in protecting the genetic material of cells, as DNA damage levels were low, with no difference between control and Me2SO10% + A, dilutions MTG10%+C, Me2SO10%+B and Me2SO15%+B.(AU)

O objetivo deste estudo foi determinar as características do sêmen de Astyanax lacustris após indução hormonal e avaliar a sensibilidade dos espermatozoides da espécie a soluções crioprotetoras baseadas nos crioprotetores dimetilsulfóxido e metilglicol. Volume, cor, concentração espermática, motilidade total e aspectos do movimento espermático foram analisados usando o "Sistema Integrado de Análise de Sêmen (ISAS®CASA)". Três extensores diferentes foram testados: A) glicose 5%+gema de ovo 10%, B) BTS® 5% e C) glicose 5% e dois crioprotetores permeáveis: dimetilsulfóxido (Me2SO) e metilglicol (MTG). O sêmen fresco de A. lacustris apresentou motilidade total 76,6±11,2%, duração da motilidade 33,0±2,2s, concentração de espermatozoides 7,22±3,2×109sptz/mL e osmolalidade seminal 219±0,03mOsm/kg-1. O teste de toxicidade apresentou maiores valores de motilidade total nas diluições MTG15%+A, Me2SO15%+B e Me2SO10%+C, e as diluições Me2SO10%+C e Me2SO15%+C apresentaram maiores valores de velocidade curvilínea, velocidade linear e velocidade média. O protocolo testado não foi eficaz em manter a viabilidade do sêmen de A. lacustris pós-congelamento, pois não foi observada motilidade em nenhuma das diluições. No entanto, o Ensaio Cometa demonstrou que as soluções crioprotetoras eram eficazes na proteção do material genético das células, pois os níveis de dano ao DNA eram baixos, sem diferença entre controle e Me2SO10%+A, MTG10%+C, Me2SO10%+B e Me2SO15%+B.(AU)

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This study aimed to determine the semen characteristics of Astyanax lacustris after hormonal induction and to evaluate the sensitivity of the species sperm to cryoprotective solutions based on the cryoprotectants dimethyl sulfoxide and methyl glycol. Volume, color, sperm concentration, total motility and aspects of sperm movement were analyzed using "Integrated Semen Analysis System". Three different extenders were tested: A) glucose 5%+egg yolk 10%, B) BTS®5% and C) glucose 5% and two permeable cryoprotectants: dimethyl sulfoxide (Me2SO) and methyl glycol (MTG). Fresh A. lacustris semen presented total motility of 76.6±11.2%, motility duration of 33.0±2.2s, sperm concentration of 7.22±3.2×109sptz/mL and seminal osmolality of 219±0.03mOsm/kg-1. The toxicity test showed the highest total motility values at the MTG15%+A, Me2SO15%+B and Me2SO10%+C dilutions, and the Me2SO10%+C and Me2SO15%+C dilutions presented the highest values for curvilinear velocity, linear velocity and average velocity. The tested protocol was not effective at maintaining the viability of A. lacustris semen after freezing because no motility was observed in any of the dilutions. However, the Comet Assay demonstrated that cryoprotectant solutions were effective in protecting the genetic material of cells, as DNA damage levels were low, with no difference between control and Me2SO10% + A, dilutions MTG10%+C, Me2SO10%+B and Me2SO15%+B.(AU)

O objetivo deste estudo foi determinar as características do sêmen de Astyanax lacustris após indução hormonal e avaliar a sensibilidade dos espermatozoides da espécie a soluções crioprotetoras baseadas nos crioprotetores dimetilsulfóxido e metilglicol. Volume, cor, concentração espermática, motilidade total e aspectos do movimento espermático foram analisados usando o "Sistema Integrado de Análise de Sêmen (ISAS®CASA)". Três extensores diferentes foram testados: A) glicose 5%+gema de ovo 10%, B) BTS® 5% e C) glicose 5% e dois crioprotetores permeáveis: dimetilsulfóxido (Me2SO) e metilglicol (MTG). O sêmen fresco de A. lacustris apresentou motilidade total 76,6±11,2%, duração da motilidade 33,0±2,2s, concentração de espermatozoides 7,22±3,2×109sptz/mL e osmolalidade seminal 219±0,03mOsm/kg-1. O teste de toxicidade apresentou maiores valores de motilidade total nas diluições MTG15%+A, Me2SO15%+B e Me2SO10%+C, e as diluições Me2SO10%+C e Me2SO15%+C apresentaram maiores valores de velocidade curvilínea, velocidade linear e velocidade média. O protocolo testado não foi eficaz em manter a viabilidade do sêmen de A. lacustris pós-congelamento, pois não foi observada motilidade em nenhuma das diluições. No entanto, o Ensaio Cometa demonstrou que as soluções crioprotetoras eram eficazes na proteção do material genético das células, pois os níveis de dano ao DNA eram baixos, sem diferença entre controle e Me2SO10%+A, MTG10%+C, Me2SO10%+B e Me2SO15%+B.(AU)

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We aimed to vitrify embryos of Prochilodus lineatus in a high-osmolarity cryoprotectant solution, evaluating, after the vitrification-thawing process, their morphological changes. Thus, 240 embryos in the 20-somite phase (20S) were exposed for 20 min to one main internal cryoprotectant solution (1,2-propanediol-PROP), divided into four immersion sequence steps of five minutes each. The first three steps were performed in solutions containing only a main internal cryoprotectant (PROP-2, 3 and 4 M), and the fourth step in a high-osmolarity solution combining internal (PROP + dimethyl sulphoxide-Me2 SO) and external cryoprotectants (sucrose-SUC). The final concentration of vitrification was PROP 5 M + Me2 SO 5 M + SUC 0.2 M. During vitrification, the straws exhibited a translucent solid appearance; however, during thawing, their structure became totally opaque and white. After thawing, the embryos suffered an increase in volume and presented morphological changes including protrusions on the surface of the yolk sac, yolk sac rupture, and optical vesicle degradation. On the inside, we observed intercellular spaces and a yolk syncytial layer (YSL) with altered chromatin. Yet, structures such as somites, neural tube, endoderm and epidermis presented cells with a nucleus and integral mitochondria. We conclude that the use of the tested cryoprotectant solution permits the formation of a vitreous solid and preserves part of the cells of the blastoderm. Yet, the heating protocol does not control recrystallization, resulting in the formation of serious morphological anomalies that prevent the preservation of the embryonic unit.

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SummaryThis study aimed to understand how germ cell development occurs in females of Devario aequipinnatus, by morphologically describing oogenesis and the reproductive phases. Sexually mature females of D. aequipinnatus (n = 70) were obtained from commercial fisheries and delivered to the Laboratório de Ictiologia Neotropical, UNESP, Ilha Solteira, SP, Brazil. The ovaries were removed, fragmented and fixed following the usual techniques for light microscopy. The stages of ovarian development in D. aequipinnatus begin with the oogonia, which proliferate into new cells or differentiate into prophasic oocytes that, at the end of this process, form the ovarian follicle and end folliculogenesis. In the previtellogenic stage, the oocytes were characterized mainly by the gradual loss of basophilia and an increase in oocyte diameter. Vitellogenesis was marked mainly by the incorporation of yolk granules. Mature oocytes were defined by their migration from the nucleus to the micropyle. Postovulatory follicles and atresic oocytes were also observed. The reproductive phases were classified as: immature, early and final developing, spawning capable, regressing and regenerating. Therefore, the development of an understanding of cell modifications that occurs up to oogenesis is a basic step that is essential for the description of the reproductive biology of D. aequipinnatus, given the lack of information about the reproductive aspects of this species.

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This study evaluated the effect of the cryoprotectants and the low temperatures on the embryonic development of Prochilodus lineatus, describing their main morphological alterations. On chilling sensitivity test, the survival rates at the twenty somites stage (20S) were 53.6% at 0ºC, and 100% in 5ºC. To test toxicity, the embryos were exposed to a graded series of 1,2-Propanediol (PROP), dimethyl sulfoxide (Me2SO4) and glycerol (GLY), terminating in a solution of high osmolarity. There was no significant difference in the embryos survival of toxicity test between series of PROP and Me2SO4 in the 6S and 20S. In the cooling protocols, were evaluated the effects of low temperature associated with cryoprotectants. At 5ºC, PROP showed survival rates above 75% in the gastrula stage (G) and above 90% in the 6S and 20S stages. High rates of abnormalities were observed, and the most recurrent were: small bodies, fins presenting uncontrolled cell growth, membrane rupture, and retraction. These results demonstrate the need to use cryoprotectant solutions, even when there is no ice nucleation, and, on the other hand, shows that high cryoprotectant concentrations promote numerous morphological lesions, compromising normal embryonic development.(AU)

Este estudo avaliou os efeitos dos crioprotetores e das baixas temperaturas no desenvolvimento embrionário de Prochilodus lineatus, descrevendo suas principais alterações morfológicas. Nos testes de sensibilidade ao frio, as taxas de sobrevivência, no estágio de vinte somitos (20S), foram de 53,6% a 0ºC e 100% a 5ºC. Nos testes de toxicidade, os embriões foram expostos a uma série graduada dos crioprotetores: 1,2-Propanediol (PROP), dimetilsulfóxido (Me2SO4) e glicerol (GLY), terminando em uma combinação de alta osmolaridade. Não houve diferença significativa na sobrevivência dos embriões nas séries do PROP e Me2SO4 nos estágios de 6S e 20S. Nos protocolos de resfriamento, foram avaliados os efeitos da baixa temperatura associados as séries crioprotetoras. A 5ºC, o PROP apresentou taxas de sobrevivência acima de 75% no estádio de gástrula (G) e acima de 90% nos estádios 6S e 20S. Foram observadas altas taxas de anormalidades sendo as mais recorrentes: corpos pequenos, nadadeiras apresentando crescimento celular descontrolado, ruptura da membrana e retração vitelínica. Estes resultados demonstram a necessidade do uso das soluções crioprotetoras, mesmo não havendo nucleação de gelo e, em contrapartida, demonstra que as elevadas concentrações crioprotetoras promovem numerosas lesões morfológicas, comprometendo o desenvolvimento embrionário.(AU)

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This study evaluated the effect of the cryoprotectants and the low temperatures on the embryonic development of Prochilodus lineatus, describing their main morphological alterations. On chilling sensitivity test, the survival rates at the twenty somites stage (20S) were 53.6% at 0ºC, and 100% in 5ºC. To test toxicity, the embryos were exposed to a graded series of 1,2-Propanediol (PROP), dimethyl sulfoxide (Me2SO4) and glycerol (GLY), terminating in a solution of high osmolarity. There was no significant difference in the embryos survival of toxicity test between series of PROP and Me2SO4 in the 6S and 20S. In the cooling protocols, were evaluated the effects of low temperature associated with cryoprotectants. At 5ºC, PROP showed survival rates above 75% in the gastrula stage (G) and above 90% in the 6S and 20S stages. High rates of abnormalities were observed, and the most recurrent were: small bodies, fins presenting uncontrolled cell growth, membrane rupture, and retraction. These results demonstrate the need to use cryoprotectant solutions, even when there is no ice nucleation, and, on the other hand, shows that high cryoprotectant concentrations promote numerous morphological lesions, compromising normal embryonic development.(AU)

Este estudo avaliou os efeitos dos crioprotetores e das baixas temperaturas no desenvolvimento embrionário de Prochilodus lineatus, descrevendo suas principais alterações morfológicas. Nos testes de sensibilidade ao frio, as taxas de sobrevivência, no estágio de vinte somitos (20S), foram de 53,6% a 0ºC e 100% a 5ºC. Nos testes de toxicidade, os embriões foram expostos a uma série graduada dos crioprotetores: 1,2-Propanediol (PROP), dimetilsulfóxido (Me2SO4) e glicerol (GLY), terminando em uma combinação de alta osmolaridade. Não houve diferença significativa na sobrevivência dos embriões nas séries do PROP e Me2SO4 nos estágios de 6S e 20S. Nos protocolos de resfriamento, foram avaliados os efeitos da baixa temperatura associados as séries crioprotetoras. A 5ºC, o PROP apresentou taxas de sobrevivência acima de 75% no estádio de gástrula (G) e acima de 90% nos estádios 6S e 20S. Foram observadas altas taxas de anormalidades sendo as mais recorrentes: corpos pequenos, nadadeiras apresentando crescimento celular descontrolado, ruptura da membrana e retração vitelínica. Estes resultados demonstram a necessidade do uso das soluções crioprotetoras, mesmo não havendo nucleação de gelo e, em contrapartida, demonstra que as elevadas concentrações crioprotetoras promovem numerosas lesões morfológicas, comprometendo o desenvolvimento embrionário.(AU)

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The present study describes the testicular structure, spermatogenesis and maturation phases in the Devario aequipinnatus, a species with great potential for use as a biological model for animal experimentation. The testes in D. aequipinnatus were classified as tubular anastomosing with unrestricted spermatogonial distribution. Were observed four phases of testicular maturation based on the changes of the testicular germinal epithelium associated with the stages of germ cells present: Early Maturation, Mid Maturation, Late Maturation and Regression. Due to the phylogenetic proximity, testicular morphology D aequipinnatus is similar to other Cyprinídeos, example, zebrafish (Danio rerio), which corroborates the hypothesis that the use of D aequipinnatus as a biological model for animal experimentation.(AU)

O presente estudo descreve a estrutura, fases de maturação testicular e espermatogênese de Devario aequipinnatus, uma espécie com grande potencial de utilização como modelo biológico em experimentação animal. D. aequipinnatus possui o testículo tubular anastomosado com distribuição espermatogonial irrestrita. Foram observadas quatro fases de maturação testicular, com base nas alterações do epitélio germinativo testicular associado aos estágios das células germinativas presentes: Maturação Inicial, Maturação Intermediária, Maturação Final e Regressão. Devido à proximidade filogenética, a morfologia testicular do D aequipinnatus é semelhante aos demais Cyprinídeos, a exemplo, zebrafish (Danio rerio), o que corrobora com a hipótese da utilização do D aequipinnatus como modelo biológico em experimentação animal.(AU)

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The present study describes the testicular structure, spermatogenesis and maturation phases in the Devario aequipinnatus, a species with great potential for use as a biological model for animal experimentation. The testes in D. aequipinnatus were classified as tubular anastomosing with unrestricted spermatogonial distribution. Were observed four phases of testicular maturation based on the changes of the testicular germinal epithelium associated with the stages of germ cells present: Early Maturation, Mid Maturation, Late Maturation and Regression. Due to the phylogenetic proximity, testicular morphology D aequipinnatus is similar to other Cyprinídeos, example, zebrafish (Danio rerio), which corroborates the hypothesis that the use of D aequipinnatus as a biological model for animal experimentation.

O presente estudo descreve a estrutura, fases de maturação testicular e espermatogênese de Devario aequipinnatus, uma espécie com grande potencial de utilização como modelo biológico em experimentação animal. D. aequipinnatus possui o testículo tubular anastomosado com distribuição espermatogonial irrestrita. Foram observadas quatro fases de maturação testicular, com base nas alterações do epitélio germinativo testicular associado aos estágios das células germinativas presentes: Maturação Inicial, Maturação Intermediária, Maturação Final e Regressão. Devido à proximidade filogenética, a morfologia testicular do D aequipinnatus é semelhante aos demais Cyprinídeos, a exemplo, zebrafish (Danio rerio), o que corrobora com a hipótese da utilização do D aequipinnatus como modelo biológico em experimentação animal.

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We aimed to analyze the oogenesis of adult females of the cichlid fish Laetacara araguaiae. The specimens' gonads were removed and processed for light and transmission electron microscopy. Oogenesis in L. araguaiae showed the following characteristics: a germinal epithelium with three types of oogonia (A-undifferentiated, A-differentiated and B-oogonia), oocytes at meiotic prophase stage and ovarian follicle formation. Oocytes showing primary growth with pre-vitellogenic and cortical alveolus were observed. Similar to data for other cichlids, oocytes in secondary growth or vitellogenesis were characterized by the initial deposition of yolk microgranules. The event that characterizes the maturation stage is nucleolus migration, also called the germinal vesicle, to the oocyte periphery in the direction of the micropyle. The follicular complex undergoes several changes throughout the oocyte stages. To the best of our knowledge this study is the first to describe L. araguaiae oogenesis. Moreover, this study is the first step to better understand the reproductive biology of this species, which shows great potential for use as an ornamental fish.

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This study aimed to establish a hormonal induction protocol for spermiation of Brycon cephalus males, using Ala6, Pro9Net-mGnRH + metoclopramide (Ovopel®). Thus, 20 males were used divided into three inductor treatments [⅓ pellet/kg (T1), ⅔ pellet/kg (T2) and 1⅓ pellet/kg (T3)] and one control group (CO), which only received physiological solution applications (0.9% NaCl). All treatments were applied in a single dose. For evaluation of the availability of the treatment, the following seminal parameters were analyzed: seminal volume, subjective spermatic motility, duration of motility, pH, osmolality and spermatic concentration. T3 showed the highest seminal volume (4.66 ± 1.52 ml), and was significantly different in comparison with T1 (2.0 ± 0.9 ml), T2 (3.5 ± 1.3 ml) and CO (2.3 ± 1.2 ml). In relation to spermatic motility, T2 and T3 showed significantly higher levels [5, (81-100%)]. However, T3 showed significantly lower average sperm motility duration than T1, T2 and CO (30 ± 7 s; 28 ± 6 s; 32 ± 8 s, respectively). With regard to the seminal parameters of spermatic concentration, pH and osmolality, no significant variation was verified among treatments. In conclusion, mGnRH + metoclopramide used for hormonal induction of B. cephalus reproduction does not induce changes related to spermatic concentration, pH and osmolality parameters of the seminal fluid and the most adequate doses among tested treatments were ⅔ pellet/kg live fish.

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RESUMO

This study aimed to detailed description of the characteristics of the different germ cell types found in Astyanax altiparanae during spermatogenesis. In this purpose, testes from 25 adult males specimens of A. altiparanae were sampled and submitted to the usual techniques for light microscopy. Based on nuclear shape, chromatin condensation, nucleoli quantity and cell size were identified four spermatogonial types: type A undifferentiated (Aund.*); type A undifferentiated (Aund.); type A differentiated (Adif.); and type B spermatogonia. Spermatocytes were observed in different phases of meiosis (leptotene/zygotene/pachytene and diplotene), metaphase I and II and secondary spermatocytes, being distinguish mainly by their chromosomal organization inside the nucleus. Were also identified three different types of spermatids, which were named as initial, intermediate and final, which can be differentiated by increase in nuclear condensation and spacing among cells inside the cysts, possibly by flagella arise and reduction in nuclear diameter. Thus, this study contribute to a better understand of spermatogenesis in this and other fish species.

O presente estudo teve como objetivo a descrição detalhada dos diferentes tipos de células germinativas encontradas em Astyanax altiparanae durante a espermatogênese. Deste modo, os testículos de 25 espécimes machos e adultos de A. altiparanae foram coletados e submetidos às técnicas usuais para microscopia de luz. Baseado no formato nuclear, condensação da cromatina, quantidade de nucléolos e tamanho celular foram identificados quatro tipos de espermatogônias: indiferenciadas do tipo A (Aund.*); indiferenciadas do tipo A (Aund.); diferenciadas do tipo A (Adif.); e espermatogônia do tipo B. Os espermatócitos foram observados em diferentes fases da meiose (leptóteno/zigóteno, paquíteno e diplóteno), metáfase I e II e espermatócitos secundários, sendo distinguidos principalmente pela organização dos cromossomos nos núcleos. Também foram identificados três diferentes tipos de espermátides, que foram nomeadas como iniciais, intermediárias e finais, que se diferenciaram pelo aumento da compactação nuclear e espaçamento entre as células no cisto, pelo possível surgimento do flagelo, e pelo diâmetro nuclear. Assim, este estudo contribui para um melhor entendimento da espermatogênese nesta e nas demais espécies de peixes.

Assuntos

RESUMO

This study aimed to detailed description of the characteristics of the different germ cell types found in Astyanax altiparanae during spermatogenesis. In this purpose, testes from 25 adult males specimens of A. altiparanae were sampled and submitted to the usual techniques for light microscopy. Based on nuclear shape, chromatin condensation, nucleoli quantity and cell size were identified four spermatogonial types: type A undifferentiated (Aund.*); type A undifferentiated (Aund.); type A differentiated (Adif.); and type B spermatogonia. Spermatocytes were observed in different phases of meiosis (leptotene/zygotene/pachytene and diplotene), metaphase I and II and secondary spermatocytes, being distinguish mainly by their chromosomal organization inside the nucleus. Were also identified three different types of spermatids, which were named as initial, intermediate and final, which can be differentiated by increase in nuclear condensation and spacing among cells inside the cysts, possibly by flagella arise and reduction in nuclear diameter. Thus, this study contribute to a better understand of spermatogenesis in this and other fish species.(AU)

O presente estudo teve como objetivo a descrição detalhada dos diferentes tipos de células germinativas encontradas em Astyanax altiparanae durante a espermatogênese. Deste modo, os testículos de 25 espécimes machos e adultos de A. altiparanae foram coletados e submetidos às técnicas usuais para microscopia de luz. Baseado no formato nuclear, condensação da cromatina, quantidade de nucléolos e tamanho celular foram identificados quatro tipos de espermatogônias: indiferenciadas do tipo A (Aund.*); indiferenciadas do tipo A (Aund.); diferenciadas do tipo A (Adif.); e espermatogônia do tipo B. Os espermatócitos foram observados em diferentes fases da meiose (leptóteno/zigóteno, paquíteno e diplóteno), metáfase I e II e espermatócitos secundários, sendo distinguidos principalmente pela organização dos cromossomos nos núcleos. Também foram identificados três diferentes tipos de espermátides, que foram nomeadas como iniciais, intermediárias e finais, que se diferenciaram pelo aumento da compactação nuclear e espaçamento entre as células no cisto, pelo possível surgimento do flagelo, e pelo diâmetro nuclear. Assim, este estudo contribui para um melhor entendimento da espermatogênese nesta e nas demais espécies de peixes.(AU)

Assuntos

RESUMO

The histological description of the urogenital papilla is an important tool to comprehension of the reproductive mechanisms in fish, as well as a pre-requisite to germ cell transplantation in adult fish, besides to be a good biological indicator to environmental changes. Was performed the histological description of the urogenital papilla and its component ducts in the tetra Astyanax altiparanae. The genital and urinay ducts pass separately throughout most part of its extension, joining in a single duct before opening. In males this opening is asymmetric and seems to have double origin, being completely surrounded by striated muscle fibers, while in females it is symmetric and the muscle fibers does not surround it totally. Spermatic duct and oviduct undergo changes throughout their extension, mainly in the morphology of the surrounding epithelium. In the spermatic duct, squamous epithelial cells change to columnar and cuboid with possible secretory activity, close to testes. In the oviduct, anteriorly epithelial cells are also squamous, however, close to ovary there are lamellae composed by a pseudostratified epithelium with columnar and cuboid cells. The urinary duct is highly similar for both sexes presenting globoid cells, which description is known in mammals, however, rare in fish.(AU)

A descrição histológica da papila urogenital é uma importante ferramenta para a compreensão dos mecanismos reprodutivos em peixes, assim como um pré-requisito para a realização do transplante de células germinativas em peixes adultos, além de um bom indicador biológico de possíveis alterações ambientais. Foi realizada a investigação histológica da papila urogenital e seus ductos constituintes no lambari Astyanax altiparanae. Os ductos genital e urinário ocorrem separadamente ao longo de maior parte de sua extensão, entretanto, unem-se em um ducto simples antes de abrir para o meio externo. Nos machos esta abertura é assimétrica e parece ter dupla origem, sendo completamente envolvida por fibras musculares estriadas, enquanto nas fêmeas ela é simétrica e as fibras musculares não a envolve totalmente. O ducto espermático e oviducto sofrem alterações ao longo de sua extensão, principalmente na morfologia do epitélio que os envolve. No ducto espermático as células epiteliais passam de pavimentosas a colunares e cuboides, com possível atividade secretora, à medida que se aproxima dos testículos. No oviducto, anteriormente as células também são epiteliais pavimentosas, entretanto, próximo aos ovários, formam-se lamelas compostas por um epitélio pseudoestratificado composto por células cuboides e colunares. O ducto urinário é bastante similar em ambos os sexos apresentando células globosas, cuja descrição é conhecida em mamíferos, porém rara em peixes.(AU)

Assuntos

RESUMO

The histological description of the urogenital papilla is an important tool to comprehension of the reproductive mechanisms in fish, as well as a pre-requisite to germ cell transplantation in adult fish, besides to be a good biological indicator to environmental changes. Was performed the histological description of the urogenital papilla and its component ducts in the tetra Astyanax altiparanae. The genital and urinay ducts pass separately throughout most part of its extension, joining in a single duct before opening. In males this opening is asymmetric and seems to have double origin, being completely surrounded by striated muscle fibers, while in females it is symmetric and the muscle fibers does not surround it totally. Spermatic duct and oviduct undergo changes throughout their extension, mainly in the morphology of the surrounding epithelium. In the spermatic duct, squamous epithelial cells change to columnar and cuboid with possible secretory activity, close to testes. In the oviduct, anteriorly epithelial cells are also squamous, however, close to ovary there are lamellae composed by a pseudostratified epithelium with columnar and cuboid cells. The urinary duct is highly similar for both sexes presenting globoid cells, which description is known in mammals, however, rare in fish.

A descrição histológica da papila urogenital é uma importante ferramenta para a compreensão dos mecanismos reprodutivos em peixes, assim como um pré-requisito para a realização do transplante de células germinativas em peixes adultos, além de um bom indicador biológico de possíveis alterações ambientais. Foi realizada a investigação histológica da papila urogenital e seus ductos constituintes no lambari Astyanax altiparanae. Os ductos genital e urinário ocorrem separadamente ao longo de maior parte de sua extensão, entretanto, unem-se em um ducto simples antes de abrir para o meio externo. Nos machos esta abertura é assimétrica e parece ter dupla origem, sendo completamente envolvida por fibras musculares estriadas, enquanto nas fêmeas ela é simétrica e as fibras musculares não a envolve totalmente. O ducto espermático e oviducto sofrem alterações ao longo de sua extensão, principalmente na morfologia do epitélio que os envolve. No ducto espermático as células epiteliais passam de pavimentosas a colunares e cuboides, com possível atividade secretora, à medida que se aproxima dos testículos. No oviducto, anteriormente as células também são epiteliais pavimentosas, entretanto, próximo aos ovários, formam-se lamelas compostas por um epitélio pseudoestratificado composto por células cuboides e colunares. O ducto urinário é bastante similar em ambos os sexos apresentando células globosas, cuja descrição é conhecida em mamíferos, porém rara em peixes.

Assuntos