Larval development of Brycon amazonicus (Teleostei, Bryconidae) with a focus on locomotory, respiratory and feeding structures.

In the present work we describe the larval development of Brycon amazonicus, with emphasis on structures linked to swimming, respiration and feeding. We monitored from larval hatching (13 h post-fertilization) to juvenile stage. Formation of the oral cavity and differentiation of gill arches began within 4 h post-hatching (hph) and within 11 hph the primordium of the pectoral fin appeared. At 25 hph, the head was in a rectilinear position with a terminal mouth, when the onset of tooth formation appeared. At 35 hph, gill arches were covered by the operculum and branchiostegal membrane and cusps of teeth pierced the epithelium of the premaxilla and anterior dentary region. Sharp teeth were observed in the maxilla at 171 hph and the pelvic fin primordium appeared at 243 hph, with taste buds on the tongue and the roof of the buccopharyngeal cavity. At 579 hph, all structures related to swimming and food capture were formed. Exogenous feeding of the larvae started at 29 hph, before the endogenous reserves were used up and the development of efficient swimming ability, reflecting the species' need to increasingly specialize on prey capture.

Efecto de la restricción de alimento y posterior realimentación sobre algunas variables fisiológicas y metabólicas en yamú (Brycon amazonicus) / Effects of food restriction and refeeding on some physiological and metabolic variables of yamú (Brycon amazonicus)

RESUMEN Se evaluó el efecto de la restricción de alimento y posterior realimentación sobre algunas variables fisiológicas en yamú (Brycon amazonicus) utilizando 1115 juveniles (peso inicial 217,75 g ± 14,74 g) distribuidos en tres estanques y asignados a los siguientes tratamientos: 1) Control (C), alimentación todos los días; 2) Restricción moderada (RM), restricción por 2 días y realimentación por 3 y 3) Restricción severa (RS), restricción total durante 5 semanas y realimentación a partir de la semana 6 hasta la 12. Se realizaron colectas de sangre los días 1, 35, 42, 56, 70 y 84, de 7 animales por tratamiento para determinación del hematocrito, proteína, glucosa, lactato, triglicéridos, colesterol, cortisol, T3 e insulina. Adicionalmente, los animales se sacrificaron para retirar el hígado y determinar el glucógeno hepático. El modelo experimental fue mixto de efectos fijos, con 3 tratamientos y 6 muestreos, con anidamiento del muestreo en los tratamientos. Los datos se analizaron a través de análisis de varianza y en los casos en que hubo diferencias significativas se empleó la prueba de Tukey-Kramer. No hubo diferencias significativas (p > 0,05) para hematocrito, proteína, glucosa, cortisol y lactato entre los tratamientos en ninguna de las muestras tomadas. La insulina y T3 mostraron diferencias entre tratamientos en el día 35, pero no en el 84, igual a lo ocurrido con colesterol y glucógeno hepático. Los triglicéridos mostraron diferencias a lo largo del estudio, en el día 84 RS presentó menores niveles. Los resultados permiten concluir que pese a la restricción parcial de alimento, no se generaron efectos negativos sobre el metabolismo de Brycon amazonicus.

ABSTRACT The effect of food restriction and refeeding on some physiological variables in yamú (Brycon amazonicus) was evaluated using 1115 juveniles (initial weight 217.75 g ± 14.74 g), distributed in three ponds. The groups were randomly assigned one of the following treatments: 1) Control (C), fed every day; 2) Moderate restriction (MR), restriction during 2 days and refeeding during 3; and 3) Severe restriction (SR), total restriction during 5 weeks and refeeding from week 6 to 12. Blood samples from 7 animals from each treatment were taken on days 1, 35, 42, 56, 70, and 84 to determine hematocrit, proteins, glucose, lactate, triglycerides, cholesterol, cortisol, T3, and insulin. The animals were sacrificed and their livers removed to calculate the hepatic glycogen. The experimental design used was a fixed effects mixed model of 3 treatments and 6 samples, with sampling nesting in the treatments. A variance analysis was performed to analyze the data; the Tukey-Kramer test a variance analysis was used in the cases with significant differences. No significant differences (p> 0.05) were found for hematocrit, protein, glucose, cortisol, and lactate. The insulin and T3 showed differences among treatments on day 35, but not on day 84, the same as cholesterol and hepatic glycogen. Triglycerides showed differences throughout the study, SR showed the lowest levels on day 84. The results allow us to conclude that, despite the partial feed restriction, there were no adverse effects on the metabolism of Brycon amazonicus.

Microcystin-LR leads to oxidative damage and alterations in antioxidant defense system in liver and gills of Brycon amazonicus (SPIX & AGASSIZ, 1829).

Microcystin's (MCs) are toxins produced by several groups of cyanobacteria, in water bodies throughout the world, in a process which is being intensified by human action. Among the variants of MCs, MC-LR stands out for its distribution and toxicity. MCs are potent inhibitors of protein phosphatases 1 and 2 A, which causes disruption of the cytoskeleton and consequent cell death. They can also alter the antioxidant system and induce oxidative stress in various organs of many species. There is, however, a lack of information about the effects of MCs on the antioxidant system and oxidative damage in Brazilian fishes. This study evaluated the effect of microcystin-LR on the antioxidant system in liver and gills of the Brazilian fish Brycon amazonicus, after 48 h of i.p injection of 100 µg MC-LR.kg-1 body mass. The liver exhibited increases in the activity of GST (74%) and GPx (217%), and a 47% decrease in SOD activity, with no changes in CAT values. In the gills of fish exposed to MC-LR, CAT and GPx activities did not show significant changes, while SOD and GST activity decreased by 66% and 37%, respectively. The GSH content did not change significantly in the liver, however, a decrease of 43% was observed in the gills. Oxidative damage measured by protein oxidation (PC) and lipoperoxidation (LPO) showed significant effects in both tissues. In hepatic tissue, there was no change in PC levels but LPO increased by 116%. Conversely, in the gills LPO levels did not change but PC increased by 317%. In conclusion, these data show that MC-LR induces oxidative damage in both tissues but in different ways, with being liver most sensitive to LPO and gills to PC. This also suggests that the gills are most sensitive to oxidative stress than liver, due to the inhibition of its antioxidant responses following MC-LR exposure.

Effects of exercise training on excitation-contraction coupling, calcium dynamics and protein expression in the heart of the Neotropical fish Brycon amazonicus.

Matrinxã (Brycon amazonicus) is a great swimming performance teleost fish from the Amazon basin. However, the possible cardiac adaptations of this ability are still unknown. Therefore, the aim of the present work was to investigate the effects of prolonged exercise (EX group - 60days under 0.4BL·s-1) on ventricular contractility by (i) in-vitro analysis of contractility comparing the relative roles of sodium/calcium exchanger (NCX) and sarcoplasmic reticulum (SR) in the excitation-contraction (E-C) coupling and (ii) molecular analysis of NCX, sarcoplasmic reticulum Ca2+ ATPase (SERCA2) and phospholamban (PLB) expression and quantification. The exercise training significantly improved twitch tension, cardiac pumping capacity and the contraction rate when compared to controls (CT). Inhibition of the NCX function, replacing Na+ by Li+ in the physiological solutions, diminished cardiac contractility in the EX group, reduced all analyzed parameters under both high and low stimulation frequencies. The SR blockage, using 10µM ryanodine, caused ~50% tension reduction in CT at most analyzed frequencies while in EX, reductions (34-54%) were only found at higher frequencies. SR inhibition also decreased contraction and relaxation rates in both groups. Additionally, higher post-rest contraction values were recorded for EX, indicating an increase in SR Ca2+ loading. Higher NCX and PLB expression rates and lower SERCA2 rates were found in EX. Our data indicate that matrinxã presents a modulation in E-C coupling after exercise-training, enhancing the SR function under higher frequencies. This was the first study to functionally analyze the effects of swimming-induced exercise on fish cardiac E-C coupling.

Aloe vera bathing improved physical and humoral protection in breeding stock after induced spawning in matrinxã (Brycon amazonicus).

In this study, we show that induced spawning causes stress, an intense loss of epithelia and immunosuppression, decreasing physical and humoral protection in fish, effects that were prevented or improved in fish bathed with Aloe vera. A. vera has several medicinal properties, including wound healing and immunostimulatory effects, which we observed in this study. Fish bathed with A. vera had a higher number of epidermal goblet cells and, in general, an improved wound healing rate compared with the control after induced spawning. These effects might be related to (1) the stimulation of leukocyte activity, represented here by the increased leukocyte respiratory activity triggered by A. vera (leukocytes are recognized as playing an important role in wound repair); (2) the antimicrobial properties of A. vera, which decrease wound infection and accelerate the healing process; and (3) several mechanisms that explain the healing effect of A. vera (increased collagen synthesis, rate of epithelialization, and anti-inflammatory and moisturizing effects). Our results also suggest that caution is necessary during the induced spawning process, especially during stripping, and A. vera bathing is recommended after intensive aquaculture operations.

Spawning failure in Brycon amazonicus may be associated with ovulation and not with final oocyte maturation / A falha na desova de fêmeas de Brycon amazonicus pode estar relacionada com ovulação e não com a indução a maturação final dos ovócitos

Avaliaram-se os possíveis mecanismos envolvidos com a falha na desova de matrinxãs (Brycon amazonicus), submetidas à indução hormonal por extrato bruto de hipófise de carpa. Para tal, após a extrusão, os ovários foram coletados e analisados histomorfometricamente. Nas fêmeas que não desovaram (FNDs), a maioria dos ovócitos vitelogênicos remanescentes nos ovários atingiu a maturação final, apresentando quebra de vesícula germinativa, mas não foram ovulados (NOs). Consequentemente, estas fêmeas apresentaram frequências mais baixas de folículos pós ovulatórios (5%) quando comparadas com a que desovou (FD) (23%). Com relação aos NOs, os valores se inverteram e a frequência destes nas FNDs (21%) foi maior do que na FD (3%). Estes dados indicam que as falhas na desova desta espécie estão provavelmente relacionadas com a ovulação, uma vez que a maturação final dos ovócitos ocorre de forma similar tanto nas FNDs como na FD. Os dados sugerem que as substâncias que promovem a ovulação, como as prostaglandinas, podem aumentar o sucesso de desova em peixes reofílicos.

Velocidade crítica de natação (Ucrit) de matrinxã (Brycon amazonicus) após exposição à hipoxia / Critical swimming speed of matrinxã (Brycon amazonicus) exposed to hypoxia

A primeira resposta ao estresse é a fuga, que depende do desempenho natatório e de ajustes fisiológicos. Este estudo investigou a velocidade crítica de natação (Ucrit) de matrinxã após exposição à hipoxia. Para isso, os peixes foram expostos à hipoxia, sendo uma parte do grupo analisada antes e outra após natação forçada, por meio da Ucrit. A hipoxia resultou no aumento de lactato, glicose, cortisol e hematócrito. Mudanças nos níveis de sódio e potássio, bem como os valores de Ucrit não foram observadas. Sugere-se que o matrinxã seja sensível à hipoxia, mas os ajustes fisiológicos são suficientes para manter seu desempenho natatório.

Escape is the first response of fish to stress, that depends on the swimming performance and the physiological adjustments. This study has investigated the critical swimming speed (Ucrit) of matrinxã after exposure to hypoxia. To achieve that, the fishes were exposed to hypoxia and analyzed before and after forced swimming, using Ucrit. The hypoxia caused an increase of lactate, glucose, cortisol and hematocrit. No changes of plasma sodium and potassium levels, as well as the Ucrit, were observed. We suggest that matrinxã is sensitive to hypoxia, but the physiological adjustments are sufficient to keep its swimming performance.

Interação do exercício de natação sustentada e da densidade de estocagem no desempenho e na composição corporal de juvenis de matrinxã Brycon amazonicus / Sustained swimming and stocking density interaction in the performance and body composition of matrinxã Brycon amazonicus juveniles

O objetivo deste estudo foi estimar o efeito da densidade de estocagem associada ao exercício de natação moderada no desempenho e na composição corporal de juvenis de matrinxã (Brycon amazonicus). Foram utilizados peixes com comprimento e peso médio inicial de 12,3±0,5cm e 18,4±0,1g, os quais foram distribuídos ao acaso em dois grupos: o primeiro grupo de peixes foi estocado em três densidades correspondendo a 88, 176 e 353 peixes m-3 e foi condicionado a nadar a uma velocidade de 1,0cc s-1 (comprimento corporal por segundo) em tanques circulares de 250L, durante 70 dias. O segundo grupo de peixes foi mantido nas mesmas densidades em água parada (sem exercício) perfazendo no total seis tratamentos com três repetições. Foram estimados parâmetros de desempenho e da composição corporal, particularmente do músculo branco e do músculo vermelho de ambos os grupos. Os resultados mostraram que o exercício e a densidade afetaram significativamente o crescimento e a composição dos músculos do matrinxã. O grupo de peixes criados sob exercício moderado na densidade de 176 peixes m-3 apresentou melhor desempenho (P<0,05), que se refletiu em maior crescimento e uniformidade do peso, além de melhor fator de condição, conversão alimentar e sobrevivência. Além disso, o exercício de natação sustentada aumentou tanto a concentração de proteína no músculo branco, como a deposição de lipídios no músculo vermelho nesse mesmo grupo de peixes. Já em altas densidades de estocagem, os resultados foram contrários. Portanto, a criação de juvenis de matrinxã sob condições de exercício moderado e em densidades próximas a 176 peixes m-3 é a mais indicada, já que se foram observadas as melhores respostas de crescimento e desempenho, além de mudanças benéficas na composição corporal. Essas mudanças resultaram em melhor condicionamento físico, possibilitando, assim, maior tolerância a altas densidades de criação.

The aim of the present study was to gauge the effect of stocking density associated to the sustained swimming on the performance and body composition of matrinxã (Brycon amazonicus) juveniles. The fish were initially sized at 12.3±0.5cm length and 18.4g±0.1g weight. They were distributed randomly in two groups: the first was arranged into three densities of 88, 176 and 353 fish m-3 and conditioned to swim at 1.0BL sec-1 in circular tanks of 250L for 70 days; the second was arranged in the same fish densities but in static waters performing six treatments with three repetitions. Performance and body compositions were estimated in white and red muscles for both groups. The results express the growth and muscle composition change in response to the exercise and fish density. The fish maintained in moderate swimming at 176 fish m-3 depicted the best performance (P<0.05), reflected as best growth, weight uniformity, conditioning factor, feed conversion and survival. Otherwise, the sustained swimming enhanced the white muscle protein concentration and the red muscle lipid deposition in the same group of fish. Yet, the results were opposite in high fish densities. Therefore, the rearing of matrinxã juveniles under moderate sustained swimming and near to 176 fish m-3 is suggested, since the best responses of growth and performance, and beneficial body changes were observed. These responses resulted in better body conditioning allowing higher tolerance to high fish densities.

Comparative study on hematological parameters of farmed matrinxã, Brycon amazonicus Spix and Agassiz, 1829 (Characidae: Bryconinae) with others Bryconinae species / Estudo comparativo sobre parâmetros hematológicos de matrinxã Brycon amazonicus Spix e Agassiz, 1892(Characidae: Bryconinae) criados em cativeiro, com outras espécies de Bryconinae

The aim of this study was not only to determine the red blood cells parameters, thrombocyte and leukocyte counts in farmed Brycon amazonicus (matrinxã), to compare these parameters among Bryconinae species from literature, and also to investigate the presence of special granulocytic cells in these fish. The results of the blood cells parameters here established for farmed B. amazonicus, a species of great economic importance in Brazilian aquaculture, could help a better understanding of the blood features in natural populations of this Amazon species. Blood parameters varied between Bryconinae species investigated, mainly the red blood cell counts, hemoglobin, hematocrit and mean corpuscular volume (MCV). The presence of the blood granulocytes, neutrophils and heterophils in matrinxã suggest that both leukocytes can be a characteristic for Bryconinae family. Furthermore, it indicates that the existence of special granulocytic cells in the blood of Bryconinae species from literature is an artifact, and this was herein discussed.

O objetivo deste estudo foi determinar os parâmetros eritrocíticos, as contagens de trombócitos e de leucócitos de espécimes de Brycon amazonicus (matrinxã), criados em cativeiro e compará-los com aqueles descritos na literatura para outras espécies de Bryconinae. Além disso, foi ainda investigada a presença de células granulocíticas especiais nestes peixes. Os resultados dos parâmetros sangüíneos apresentados para B. amazonicus podem ajudar a entender melhor as características sangüíneas em população natural desta espécie de grande importância para a aqüicultura brasileira. Os parâmetros sangüíneos das espécies de Bryconinae investigadas apresentaram variação interespecíficas principalmente a contagem de eritrócitos, hemoglobina, hematócrito e volume corpuscular médio (VCM). A presença dos granulócitos sangüíneos, neutrófilos e heterófilos em matrinxã sugere que esta pode ser uma característica das espécies pertencentes à subfamília Bryconinae. Além disso, indica que a existência de células granulocíticas especiais no sangue de espécies de Bryconinae da literatura é um artefato, e isto foi aqui discutido.

Stress responses of juvenile matrinxã (Brycon amazonicus) after transport in a closed system under different loading densities / Respostas de estresse de matrinxã (Brycon amazonicus) juvenil após transporte em sistema fechado sob diferentes densidades de carga

Optimum loading density for the transport of matrinxã juveniles was investigated in a closed system with plastic bags. Transport was conducted for 4 h with fish (23.5± 0.4g and 11.6 (0.08cm) starved for 24h at loading densities of 83g L-1 (D1), 125g L-1 (D2), 168g L-1 (D3) and 206g L-1 (D4). Fish were sampled before transport (BT), after transport (AT) and 24h AT. The water quality was monitored before capturing fish in depuration tanks, after transport in plastic bags and in recovery tanks. Water oxygen decreased to values below 4mg L-1 in D2, D3 and D4, temperature was around 32°C, pH 6.5-6.78, total ammonia 1.09-1.7mg L-1, un-ionized ammonia 3.58-9.33x10³mg L-1 and alkalinity 134-165mg CaCO3 L-1. Blood cortisol and glucose concentrations increased in fish of all densities AT and recovered the BT levels 24h after arrival. Osmolality did not change AT but increased 24h AT in fish of all densities whereas plasma chloride decreased inversely to the loading densities after the procedure. Hematocrit decreased 24h AT in fish of all densities but no differences were verified in the number of erythrocytes. No mortality was registered in any treatment during the next week after transport. Matrinxã demonstrated to be a crowding tolerant-species in transport operations besides tolerating low levels of oxygen in the water.

Neste estudo, foram investigadas as densidades de carga adequadas para transporte de matrinxãs juvenis em sistema fechado com sacos plásticos. O transporte de 4h foi feito com peixes (23,5±0,4g; 11,6 (0,08cm) em jejum por 24h, em densidades de 83g L-1 (D1), 125g L-1 (D2), 168g L-1 (D3) e 206g L-1 (D4). Os peixes foram amostrados antes do transporte (AT), logo após o transporte (chegada) (DT) e 24h depois. A qualidade da água foi monitorada antes da captura dos peixes nos tanques de depuração, após o transporte nos sacos plásticos e nos tanques de recuperação. O oxigênio da água diminuiu para valores inferiores a 4mg L-1 em D2, D3 e D4, a temperatura esteve em torno de 32°C, pH 6,5-6,78, a amônia total foi de 1,09-1,7mg L-1, a amônia não-ionizada foi de 3,58-9,33 x 10³mg L-1 e alcalinidade 134-165mg CaCO3 L-1. O cortisol plasmático e a glicose sanguínea aumentaram após o transporte nos peixes em todas as densidades ensaiadas, voltando aos valores controle 24h depois. Os valores de osmolaridade não mudaram logo após o transporte, mas aumentaram 24h depois de modo igual em todas as densidades. O cloreto plasmático diminuiu na chegada, de modo inversamente proporcional à densidade de carga. O hematócrito diminuiu 24h depois da chegada dos peixes, em todas as densidades testadas, mas não houve diferença no número de eritrócitos. Não houve mortalidade até uma semana após o transporte. O matrinxã mostrou ser uma espécie tolerante a altas densidades de carga em embalagens para transporte além de suportar baixos níveis de oxigênio na água.

Metabolic responses of matrinxã to eugenol in anesthetic baths - DOI: 10.4025/actascibiolsci.v29i3.474 / Respostas metabólicas do matrinxã submetido a banhos anestésicos de eugenol - DOI: 10.4025/actascibiolsci.v29i3.474

Matrinxã (Brycon amazonicus) is a commercial fish that presents excessive movements during handling. This characteristic predisposes the animals to injuries and losses of scales that may result in high mortality rates. Eugenol, the main component of clove oil, has been reported as an alternative fish anesthetic because it is a natural product and cheap. However, studies remain necessary about the metabolic responses of tropical fishes to anesthetics. The present work evaluated metabolic responses of Brycon amazonicus to eugenol in simulated anesthetic baths. The fish metabolic demand was supplied mainly by amino acids catabolism. Typical metabolic stress responses to handling were detected but eugenol could not totally reduce them. On the other hand, the anesthetic dissolved in the water did not provoke any extra charge of stress during short-term exposures up to 60 mg L-1 for 10 min. Eugenol provides safety to the workers in handling of matrinxã.

O matrinxã (Brycon amazonicus) é uma espécie de interesse comercial. Porém, este peixe movimenta-se em excesso durante práticas de manejo, podendo sofrer ferimentos e perdas de escamas que, muitas vezes, resultam em taxas elevadas de mortalidade. O eugenol, principal componente do óleo de cravo, tem sido bastante utilizado como um anestésico alternativo para peixes por ser um produto natural e de baixo custo. Entretanto, estudos que tratam de respostas metabólicas, em peixes tropicais expostos a diferentes anestésicos, são ainda necessários. Dentro deste intuito, o presente trabalho avaliou respostas metabólicas do Brycon amazonicus ao eugenol, em simulações de banhos anestésicos. A demanda metabólica do matrinxã foi suprida principalmente pelo catabolismo de aminoácidos. Respostas típicas ao estresse foram detectadas por causa do manuseio imposto aos peixes para a realização dos banhos anestésicos. O eugenol não reduziu totalmente essas reações ao estresse. Por outro lado, esse anestésico não provocou estresse adicional em virtude de sua presença em exposições curtas de até 60 mg L-1 por 10 min. O eugenol proporciona segurança aos trabalhadores durante práticas de manejo, sem maiores prejuízos ao matrinxã.