Larvae of migratory Characiformes species in an archipelago in the Lower Amazon River

In the Amazon basin, most of the migratory Characiformes species represent an important fishing resource for local people. However, the lack of information about the main areas and periods of reproduction and the importance of certain environments for the maintenance of species may jeopardize the renewal of fish stocks. Thus, the aim was to examine the spatial and seasonal variation in the density of larvae of migratory Characiformes species in an archipelago in the Lower Amazon River and to assess the importance of this environment for the biological recruitment of the studied species. The capture of larvae was carried out in places close to restinga swamps, steep ruts,and inlet areas with a plankton net (300 μm) in monthly sampling from January to December 2013. An analysis of variance was used to verify differences in larval density between seasons and sampling months. A total of 30,997 larvae were captured and showed no significant differences in their distributions between habitats. However, a variationbetween the phases of the hydrological cycle was evident, in which the highest larval concentrations were observed during the flooding (17.72 larvae.10m-3) and the drought (1.33 larvae.10m-3). The study area has a great capacity to assist in the maintenance and renewal of regional fish stocks, as it is an important retention and nursery site for larvae of Characiformes that drift along the main channel of the Amazon River.

Effects of cryoprotectants and low temperatures on hatching and abnormal embryo development of Prochilodus lineatus (Characiformes: Prochilodontidae)

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)

Early larvae ontogeny of the Neotropical fishes: Prochilodus costatus and P. argenteus (Characiformes: Prochilodontidae) / Ontogenia temprana de larvas de peces neotropicales: Prochilodus costatus y P. argenteus (Characiformes: Prochilodontidae)

AbstractEarly development of fish larvae is a highly dynamic process and its study may provide important information about ontogenetic development, bioenergetic growth, behaviour, taxonomic characteristics for identification in natural environments, identification of spawning areas, and population monitoring. With the aim to provide knowledge about their growth and behaviour, to support larval rearing, and also taxonomic purposes, we studied the life history of the Prochilodus argenteus and P. costatus from hatching until the complete absorption of the yolk. Larvae were obtained through artificial reproduction at the Hydrobiology and Aquaculture Station of Três Marias, Minas Gerais, Brazil. Immediately after hatching, 100 larvae of each species were put in two plastic incubators for conditioning. On a daily basis, larvae behavior was recorded and 14 larvae of each species were collected to analyse body morphology. On the first day after hatching, larvae of P. costatus and P. argenteus showed an elongated and transparent body; the yolk sac was filled with individualized yolk globules. In both species, the embryonic fin rounded the caudal region of the body, the retina was non-pigmented and the gut was obliterated. At the second day post-hatching, larvae of both species dendritic chromatophores had emerged, the mouth was obliterated and the pectoral fin was observed. The larvae showed 38-43 myomeres in P. costatus and 42-43 in P. argenteus. For both species, the gas bladder was inflated and the lumen of the gut was already open. On the third day post-hatching, the mouth of P. costatus and P. argenteus was already open in a sub-terminal position; the retina was pigmented; the gill arches had lamellar protrusions and were partially covered by the operculum. On the fourth day post-hatching, the pigmentation pattern was maintained with greater intensity; the mouth occupied a terminal position, the yolk sac was almost completely reabsorbed, and the pectoral and caudal fins showed mesenchymal rays in both species. The gut showed a broad lumen with folded mucosa and epithelium with striated border. The larvae of both species showed similar swimming behaviour. Our study provided understanding about the morphophysiological aspects, species identification, larval development and growth, and the ontogenic characteristics of two Neotropical fishes with importance for commercial and sports fishing. Rev. Biol. Trop. 64 (2): 537-546. Epub 2016 June 01.

ResumenEl desarrollo temprano de las larvas de peces es un proceso altamente dinámico y estudiarlo proporciona información importante sobre su desarrollo ontogénico, su crecimiento bioenergético, su comportamiento, las características taxonómicas para la identificación en entornos naturales, la identificación de las zonas de desove y la vigilancia de la población. Los primeros años de vida de Prochilodus argenteus y Prochilodus costatus fueron estudiados desde la eclosión hasta la completa absorción de la yema, con el fin de proporcionar conocimiento sobre los caracteres taxonómicos y el crecimiento y comportamiento, lo cual puede ser aplicado al desarrollo de la cría de larvas. Las larvas fueron obtenidas junto a la Estación de Hidrobiología y Acuicultura de Três Marias, Minas Gerais, donde fueron reproducidas artificialmente. Inmediatamente después de la eclosión, aproximadamente 100 larvas de cada especie fueron acondicionadas en dos incubadoras de plástico. El comportamiento de las larvas fue registrado diariamente y fueron recolectadas 14 larvas de cada especie para análisis morfológico del cuerpo. En el primer día después de la eclosión las larvas de P. costatus y P. argenteus presentaban cuerpos alargados y transparentes. El saco vitelino se llenó de glóbulos de vitelo individualizados. En las dos especies, la aleta embrionaria circundaba la región caudal del cuerpo, la retina no presentaba pigmentos y el intestino estaba obliterado. En el segundo día, en las larvas de ambas especies habían surgido cromatóforos dendríticos, la boca estaba obliterada y fue registrada una aleta pectoral. Las larvas presentaban 38-43 miómeros en P. costatus y 42-43 en P. argenteus. En las dos ecpecies la vejiga de gas estaba inflada y el lumen del intestino ya estaba abierto. En el tercer día, la boca tanto del P. costatus como del P. argenteus ya estaba abierta en una posición subterminal. La retina estaba pigmentada, los arcos branquiales tenían protuberancias laminares y estaban parcialmente cubiertos por el opérculo. En el cuarto día, el patrón de pigmentación se presentó con mayor intensidad; la boca ocupó una posición terminal, el saco vitelino fue casi completamente reabsorbido y las aletas pectorales y caudales presentaban rayos mesenquimales en ambas especies. El intestino mostró un amplio lumen, con la mucosa plegada y el epitelio con borde estriado. Las larvas de ambas especies mostraron un comportamiento similar de natación durante el periodo de prueba. Nuestro estudio provee conocimiento sobre aspectos morfofisiológicos, identificación de especies, desarrollo y crecimiento larval y características ontogénicas de dos peces subtropicales de importancia para la pesca comercial y deportiva.

Influence of a large dam and importance of an undammed tributary on the reproductive ecology of the threatened fish matrinxã Brycon orthotaenia Günther, 1864 (Characiformes: Bryconidae) in southeastern Brazil

A total of 312 adults of matrinxã were captured bimonthly, between 2011-2012, in two sites of the rio São Francisco: immediately downstream of the Três Marias Dam (site 1) and in the confluence with the rio Abaeté (site 2). Specimens of Brycon orthotaenia from site 2 presented higher values of total length (32.24 ± 5.70 cm for females and 26.64 ± 3.79 cm for males) and body weight (506.66 ± 332.17 g for females and 267.36 ± 145.84 g for males), when compared with the site 1. In site 2 was registered the highest GSI means (9.97 ± 2.96 for females and 0.93 ± 0.52 for males), compared with the site 1 means. Reproduction period occurred from October to February. Unlike site 2, no spawned females or spent males were captured in site 1. Significant differences were found between females and males in site 2 (χ2 = 26.84, df = 1, P < 0.05). Physical-chemical parameters of the water presented higher values in site 2. The canonical correlation test indicated that the reproduction of males is less susceptible to environmental factors than that of females. These results show that B. orthotaenia finds favourable conditions to reproduction just in site 2.

Um total de 312 adultos de matrinxã foram capturados bimestralmente entre 2011-2012, em dois pontos do rio São Francisco: imediatamente a jusante da barragem de Três Marias (ponto 1) e na confluência com o rio Abaeté (ponto 2). Exemplares de Brycon orthotaenia do ponto 2 apresentaram maiores valores de comprimento total (32,24 ± 5,70 cm para fêmeas e 26,64 ± 3,79 cm para machos) e peso corporal (506,66 ± 332,17 g para fêmeas e 267,36 ± 145,84 g para machos) quando comparado com peixes do ponto 1. No ponto 2, foi registrado as maiores médias de IGS (9,97 ± 2,96 para fêmeas e 0,93 ± 0,52 para machos) em comparação com o ponto 1. O período reprodutivo ocorreu de outubro a fevereiro. Ao contrário do ponto 2, fêmeas desovadas e machos espermiados não foram capturados no ponto 1. Foram encontradas diferenças significativas entre fêmeas e machos no ponto 2 (χ2 = 26,84, gl = 1, P < 0,05). Parâmetros físico-químicos da água apresentaram maiores valores no ponto 2. A análise de correlação canônica indicou que a reprodução de machos é menos susceptível a fatores ambientais do que de fêmeas. Estes resultados mostram que B. orthotaenia encontra condições favoráveis para a reprodução apenas no ponto 2.

O desenvolvimento embrionário da Piapara, Leporinus elongatus (Pisces, Anostomidae), utilizando técnicas de histologia, microscopia eletrônica de varredura e imunológicas empregando marcadores ósseos / The embryonic development of Piapara, Leporinus elongatus (Pisces, Anostomidae), using histological techniques, electron microscopy and immunological techniques using bone markers

O desenvolvimento embrionário dos peixes é de grande importância para a piscicultura e na reintrodução de espécies ameaçadas de extinção em seus ambientes, e seu conhecimento constitui uma importante maneira para minimizar doenças e mortalidades dessas espécies. Com o auxílio de técnicas como a Microscopia Eletrônica de Varredura (MEV) e a imuno-histoquimica para identificar proteínas ósseas, foi possível avaliar as fases de desenvolvimento com mais riqueza de detalhes, facilitando a compreensão de hábitos e da biologia da espécie. Neste trabalho pudemos observar a ontogenia e osteogênese da Piapara (Leporinus elongatus), desde a fecundação até a fase juvenil, sendo evidenciadas estruturas importantes como o tamanho do vitelo, essencial para a nutrição do embrião; o fechamento do blastóporo, evento principal da embriogênese, que indica as taxas de fertilização; a metamorfose, que indica a formação dos primeiros e principais órgãos do animal e a formação de sua estrutura óssea. As Proteínas Ósseas Morfogenéticas (BMP-2 e BMP-4), moléculas essenciais reguladoras no desenvolvimento embrionário e na formação óssea, foram observadas apenas no estádio larval até o período juvenil, não sendo evidenciadas nos estágios anteriores. Os resultados desse trabalho trouxeram novas informações quanto à biologia do desenvolvimento dessa espécie, que certamente poderão auxiliar no aprimoramento de técnicas reprodutivas visando uma melhora na sua produção seja para fins comerciais ou de repovoamento.(AU)

The embryonic development of fishes has great importance in fish culture and on reintroduction of species at risk of extinction into their environment; its knowledge constitutes an important way to minimize diseases and mortality of these species. By using techniques like electron microscopy and immunohistochemistry for bone markers identification, it was possible to evaluate the developmental stages of Leporinus elongatus in more details, helping to clarify the habits and biology of this species. Results showed the ontogeny and ostheogenesis of of Leporinus elongatus this species from fecundation to juvenile, evidencing important structures as size of the yolk, essential to embryo nutrition, the blastopores closure, important event in embryogenesis because it indicates fertilization indexes, and the metamorphosis, which shows the main organs' development including bones. Bone morphogenetic markers 1 and 4, essential regulatory molecules for bone development, had their expression restricted from larva to fry stages, and were not observed in their previous stages. In sum, these results provided new data that may improve reproductive techniques for L. elongatus, and will support commercial and repopulation purposes.(AU)

Toxicity of cryoprotectants on Prochilodus lineatus (Valenciennes, 1837) (curimba) embryos in an experimental incubator (Characiformes: Prochilodontidae)

This paper investigated the effect of cryoprotectant substances on Prochilodus lineatus embryos in an experimental incubator. The prospective study applied combinations of polyvinyl alcohol, hydroxyethyl cellulose, gelatin and fetal bovine serum with dimethyl sulfoxide and ethylene glycol in a new experimental incubator. The morphology of embryos, larval viability and the efficiency of experimental incubators in maintaining the quality of embryos were evaluated. This study demonstrates the efficient association between hydroxyethylcellulose and dimethyl sulfoxide as greater viability (p<0.05) was found for embryos (72.9 ± 23.9%). It should also be noted the permeation of cryoprotectants in embryos through the changes found in chorion diameter, embryo diameter and embryo volume comparing the treatments versus control group (water) (p<0.05), this results can help in future cryopreservation protocols. Although the temperature and oxygenation differed between the usual and experimental incubators (p<0.05), the results showed a high fertilization rate (79.6 ± 13.2%) for experimental incubators (p<0.05) which is sufficient for the maintenance of embryos in a cryoprotective environment and effectively allows experimentation for long periods with cryoprotectant substances. Cryopreservation of fish embryos has not been accomplished yet and new approaches are required for understanding the permeability of teleost embryos, especially in Brazilian native species.

Este trabalho avaliou o efeito de substâncias crioprotetoras sobre embriões de Prochilodus lineatus em uma incubadora experimental. O estudo aplicou combinações de álcool polivinílico, hidroxietilcelulose, gelatina e soro fetal bovino com dimetilosulfóxido e etilenoglicol em uma nova incubadora experimental. Foram avaliadas a morfologia dos embriões, a viabilidade larval e a eficiência das incubadoras experimentais na manutenção da qualidade dos embriões. Este estudo demonstra a associação eficiente entre hidroxietilcelulose e dimetilsulfóxido pela maior viabilidade (p<0,05) encontrada para os embriões (72,9 ± 23,9%). Deve-se notar também a permeação dos crioprotetores nos embriões através das alterações encontradas no diâmetro córion, diâmetro do embrião e no volume do embrião comparando os tratamentos ao grupo controle (água) (p<0,05), estes resultados podem ajudar em futuros protocolos de criopreservação. Embora a temperatura e a oxigenação diferiram entre as incubadoras comuns e as experimentais (p<0,05), os resultados mostraram elevada taxa de fertilização (79,6 ± 13,2%) para incubadoras experimentais (p<0,05), o que é suficiente para a manutenção de embriões em ambiente crioprotetor e permite efetivamente a experimentação por longos períodos com substâncias crioprotetoras. A criopreservação de embriões de peixes ainda não foi realizada e novas abordagens são necessárias para a compreensão da permeabilidade dos embriões de teleósteos, especialmente em espécies nativas brasileiras.