Modulation of immunity and hepatic antioxidant defense by corticosteroids in pacu (Piaractus mesopotamicus).

We investigated the impact of both the oral administration of hydrocortisone (HC) and an acute stressor on stress, innate immune responses and antioxidant system/oxidative stress responses of juvenile Piaractus mesopotamicus. Fish were either 1) given a commercial feed (C), 2) given a feed supplemented with 400 mg/kg HC, or 3) fed a commercial feed, chased for 2 min and exposed to air for 4 min (S). After initial sampling, fish C and HC were fed and sampled 1, 3, 6, 24 and 72 h post-feeding. Fish S were fed at the same time as the other groups, exposed to a stressor, and sampled 1, 3, 6, 24 and 72 h after. Exposure to the stressor increased circulating glucose and cortisol levels (at 1 and 3 h, respectively), while oral HC increased circulating cortisol at 1 h and glucose at 3 h. The stressor activated respiratory activity of leukocytes (RAL) at 3 h and reduced it at 6 h. HC did not activate RAL, but it did impair it at 6 h. The serum hemolytic activity of the complement system (HAC50) was impaired by the stressor at 1 and 3 h and by HC at 1 h. Regarding the antioxidant system, exposure to the stressor reduced glutathione peroxidase (GPx) and catalase (CAT) activity and decreased concentrations of reduced glutathione (GSH) in the liver up to 6 h. HC only impaired GPx. Additionally, stress induced the accumulation of melano-macrophage (MM) and melano-macrophage centers (MMC), which are biomarkers of oxidative stress, in the spleen. Differences in biomarkers in fish given cortisol and exposed to stress indicate that exogenous hormone was unable to precisely reproduce stress responses.

Modulation of the innate immune response, antioxidant system and oxidative stress during acute and chronic stress in pacu (Piaractus mesopotamicus).

Stress is an energy-demanding process, as well as the responses of the innate immune system, that impose a metabolic overload on cellular energy production, which can affect the cellular redox balance, causing oxidative damage. We evaluated the role of stress in the modulation of innate immune and oxidative/antioxidant mechanisms in juvenile pacu exposed to acute and chronic stressors. The experimental period lasted 30 days, and fish (113.7 ± 35.1 g) were fed commercial feed. During this period, half of the fish were not manipulated (Condition A), and the other half were chased with a dip net for 5 min twice a day (Condition C). After the 30-day period, fish from both groups were sampled (baseline sampling), and the remainders (not sampled) were air exposed for 3 min (acute stressor), returned to the tanks, and were sampled again 30 min, 3 h, 6 h, and 24 h after air exposure. We evaluated biomarkers of stress (circulating cortisol and glucose), the innate immune system (respiratory burst activity/RBA, hemolytic activity of the complement system (HA-AP) and serum concentration of lysozyme), oxidative damage (lipid peroxidation/LPO), and antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; and glutathione peroxidase, GSH-Px). Our results showed that stress, acutely or chronically, caused a transient reduction of RAL and activated the HA-AP. Acutely, stress increased the lysozyme concentration. Furthermore, both conditions caused oxidative stress in the liver, and differently they modulated the antioxidant system, enhancing SOD activity and impairing CAT and GSH-Px activity.

Under stress conditions, pacu Piaractus mesopotamicus modulates the metabolic allostatic load even after Dolops carvalhoi challenge to maintain self-protection mechanisms.

Fish metabolic allostatic dynamics, when animal present physiological modifications that can be strategies to survive, are important for promoting changes to ensure whole body self-protection and survival in chronic states of stress. To determine the impact of sequential stressors on pacu (Piaractus mesopotamicus), fish were subjected to two trials of stressful treatments, administration of exogenous dietary cortisol, and parasite challenge. The first experiment consisted of a two-day acute stress trial and the second, an eight-day chronic stress trial, and after both experiments, fish parasite susceptibility was assessed with the ectoparasite Dolops carvalhoi challenge. Physiological changes in response to acute trial were observed in glycogen, cortisol, glucose, osmolarity, sodium, calcium, chloride, potassium, hematocrit, hemoglobin, red blood cells and mean corpuscular volume, and white blood cell (P < 0.05), whereas response to chronic trial were observed in glycogen, osmolarity, potassium, calcium, chloride, mean corpuscular volume, white blood cell, neutrophil, and lymphocyte (P < 0.05). Acute trials caused physiological changes, however those changes did not induce the consumption of hepatic glycogen. Chronic stress caused physiological changes that induced hepatic glycogen consumption. Under acute trial, stress experience was important to fish to achieve homeostasis after chronic stress. Changes were important to modulate the response to stressor, improve body health status, and overcome the extra stressor with D. carvalhoi challenge. The experiments demonstrate that pacu initiate strategic self-protective metabolic dynamics in acute states of stress that ensure the maintenance of important life processes in front of sequential stressors.

Physiological activity of Aloe vera in pacu (Piaractus mesopotamicus) inoculated with Aeromonas hydrophila.

Aloe vera is a traditional medicinal plant; however, its use in fish is fairly recent. We evaluated the effects of dietary A. vera on stress, innate immunity, and energy metabolism in pacu inoculated with Aeromonas hydrophila. For 7 days, 192 fish were fed with diets supplemented with 0% (control), 0.5%, 1.0%, and 2.0% of the plant extract and then inoculated with bacteria and sampled 3, 6, and 24 h later. All concentrations of A. vera reduced basal levels of cortisol, and 1.0% reduced cortisol levels more intensely 3 h after inoculation. A. vera increased the basal respiratory activity of leukocytes/RAL (0.5 and 1.0%), increased the serum levels of lysozyme (1.0 and 2.0%) 6 h after inoculation, and increased the activity of the complement system after 3 h. Spleen somatic index/SSI increased with 1.0 and 2.0% A. vera. A. vera also promoted metabolic effects. It increased basal levels of lipids in the liver and muscle, as well as hepatosomatic index (1.0%) and, 3 h after inoculation, prevented the reduction of serum triglyceride (1.0%) and reduced the mesenteric fat (1.0%). Bacterial inoculation increased RAL from 3 to 24 h and lysozyme levels at 24 h, increased serum cholesterol at 24 h, and decreased serum triglyceride from 3 to 24 h, regardless of A. vera. We concluded that A. vera offered for only 7 days had stress-reducing effects, stimulated innate immunity, protected triglyceride levels in blood, lipid depots in the liver and muscle, and directed the energy mobilization to visceral depots.

ß-Glucan enhances respiratory activity of leukocytes suppressed by stress and modulates blood glucose levels in pacu (Piaractus mesopotamicus).

We evaluated the immune response of pacu fed with a ß-glucan diet (0.5%) for 10 days. After the feeding period, fish were subjected to handling and 3 h after, inoculated with Aeromonas hydrophila. Fish were sampled before handling (baseline condition), 3, 6, and 24 h and 1 week after inoculation. A higher level of blood glucose was found in fish treated with ß-glucan in baseline conditions. Handling and bacterial inoculation increased the circulating levels of cortisol and glucose and promoted the acute inflammatory response (lymphopenia and neutrophilia). ß-Glucan prevented the decrease in the respiratory activity of leukocytes observed in the control group at 3 h sampling. ß-Glucan did not affect the complement and lysozyme, which were activated 24 h after the bacterial challenge in control fish. A reduction in the number of leukocytes was found in fish treated with ß-glucan 1 week after the challenge. We suggest two plausible hypotheses for this event: (1) it could be attributed to a depletion of the immune responses or (2) it could be due to a mobilization of the leukocytes to the spleen for antigen presenting/processing. In general, ß-glucan avoided the reduction of the activity of leukocytes after stress and the bacterial challenge and increased the baseline glucose levels. Our findings confirm the immunomodulatory action of glucan and add evidence showing that glucan can have a role in stress response.

ß-glucan modulates cortisol levels in stressed pacu (Piaractus mesopotamicus) inoculated with heat-killed Aeromonas hydrophila.

In this study, we show that ß-glucan can modulate cortisol release in fish. We simulated a common situation in aquaculture: the transport of fish followed by contact with an opportunistic pathogen and observed what effect glucan had on the immune and stress response in these conditions. Pacu (Piaractus mesopotamicus) were fed with a diet containing ß-glucan (0.1%) for 15 days prior to transport followed by an injection with heat-killed Aeromonas hydrophila. We sampled fish before transport, at arrival and at 3 and 24 h after bacterial injection. ß-Glucans are used in aquaculture and have a known immunostimulatory effect, which was observed in this study. The results showed that ß-glucan modulated the plasma cortisol levels differently by increasing these levels up to 24 h after transport and preventing the increase caused by bacterial inoculum injection. In addition, ß-glucan enhanced the activity of the complement system at 24 h and reduced the monocytes and lymphocytes number in peripheral blood at 3 and 24 h after bacterial inoculation. Our results suggest that ß-glucan modulated a bidirectional interaction between the stress and the immune responses. The modulation of cortisol levels and the immunostimulation by ß-glucan at different moments in our study suggest the compound has a protective effect by avoiding higher levels of the hormone and improving resistance against bacterial infection in pacu. These results add evidence to support the use of ß-glucan as an immunomodulator in the aquaculture industry.

Distinct ß-glucan molecules modulates differently the circulating cortisol levels and innate immune responses in matrinxã (Brycon amazonicus).

This study investigated the effects of two ß-glucan molecules with different purities and isolated by different biotechnological processes on the immune response of matrinxã (Brycon amazonicus) prior and after challenge with Aeromonas hydrophila. In this sense, we evaluated serum cortisol and plasma glucose levels, the number of leukocytes (lymphocytes, neutrophils and monocytes), as well as the respiratory activity of leukocytes prior to, 6 and 24 h post infection (hpi). During 15 days, fish were fed with diets containing 0.1% of two ß-glucans (ß-G 1 and ß-G 2, with 71 and 62% of purity, respectively) and then submitted to challenge. Results were compared with a positive control group fed with a ß-glucan-free diet. A negative control group, also fed with ß-glucan-free diet but inoculated with PBS, was established to evaluate the effect of handling during injection. Our results showed that different ß-glucans affected differently the biological responses of matrinxã. The ßG 2 modulated the cortisol profile prior to and after the acute infection with A. hydrophila, and increased the mobilization and activity of leukocytes. The infection promoted lymphopenia at 6 hpi and both ß-glucans increased the circulating lymphocyte population 24 hpi. Moreover, the ß-G 2 prevented the infection-induced neutrophilia at 6 and 24 hpi. Finally, the ß-G 2 caused a marked increase in the circulating monocytes prior to infection, and a reduction at 6 hpi that was reversed at 24 hpi. In summary, our study demonstrates that ß-G 2 was more efficient on the induction of the cell-mediate immunity in matrinxã.

Glucan-MOS® improved growth and innate immunity in pacu stressed and experimentally infected with Aeromonas hydrophila.

We tested the efficacy of a commercial product (Glucan-MOS®) derived from yeast Saccharomyces cerevisiae, containing two combined products, ß-1,3-1,6 glucans and mannans on the growth, feed efficiency, stress and innate immune responses of juvenile pacu (Piaractus mesopotamicus) after a stressful handling and bacterial inoculation. For this, we evaluated the serum cortisol and plasma glucose levels, the respiratory activity of leukocytes, the serum lysozyme levels, as well as the number of circulating erythrocytes and leukocytes of fish fed during 30 days with diets containing increased levels of Glucan-MOS (0.0, 0.1, 0.2, 0.4 and 0.8%). The supplementation of 0.1% improved weight gain, feed conversion and the protein efficiency ratio compared to a control diet. The 0.2 and 0.4% Glucan-MOS® diets were sufficient to increase the respiratory burst of leukocytes and lysozyme activity, the number of thrombocytes, neutrophils and monocytes in the blood after a stressful handling and bacterial challenge, and minimized stress response as shown by decreased cortisol and glucose levels when compared to the control. The results of this work reinforce the benefits of the adoption of feeding strategies including combination of both ß-1,3-1,6 glucans and mannans as a dietary supplement in periods prior to intensive management. The 30-day period was sufficient to stimulate growth performance, improve nutrient utilization, minimize stress response and modulate innate immunity responses.

Levamisole enhances the innate immune response and prevents increased cortisol levels in stressed pacu (Piaractus mesopotamicus).

We analyzed the effects of levamisole on stress and the innate immune responses of pacu (Piaractus mesopotamicus). A total of 300 fish (180 ± 1.27 g) were fed a diet containing levamisole hydrochloride (LHC) for 15 days, then distributed into the following groups: T0 (control group); T1 (100), T2 (150), T3 (300) and T4 (500) mg kg-1 LHC (15 fish per group and four replicates per treatment). After this, fish (n = 8 per treatment) were exposed to air for three minutes to simulate stress conditions and were then challenged with the bacterium Aeromonas hydrophila to stimulate the immune system. Fish were sampled at 1, 3 and 24 h after bacterial inoculation to measure plasma cortisol and glucose concentrations, the leukocyte respiratory burst (LRB), hemolytic activity of the complement system (HAC50) and serum lysozyme activity (SLA). LHC attenuated the increase in plasma cortisol at 1 h (500 mg kg-1) and 3 h (300 mg kg-1) after air exposure and bacterial inoculation compared to control fish. The highest glucose concentrations were observed at 1 and 3 h after stress, which then returned to initial levels after 24 h, without any effect of LHC. The LHC 100 mg kg-1 dose increased LRB 1 h after inoculation and activated the HAC50 3 h later. At 24 h, all LHC concentrations increased the HAC50. SLA was reduced after inoculation, throughout the experimental period, without an effect of levamisole. Our results indicate that the oral administration of levamisole for 15 days modulated circulating cortisol levels during the stress response and improved the innate immune response against A. hydrophila infection in pacu.

Feed restriction and insulin-like growth factor-I (IGF-I) affect the oocyte maturation in matrinxã Brycon amazonicus.

The feeding and nutrition of breeders are crucial aspects in the reproductive process. During the maturation period, metabolic changes occur aiming at mobilizing energy for growth and follicular development. The involvement of IGF-1 in metabolic and reproductive events is important. The aim of this work was to evaluate if alternate feed restriction and re-feeding have permissive effects on in vitro actions of IGF-1 on oocytes development of matrinxã. In vivo experiments were performed during vitellogenesis period. Females (n = 60) were fed with a commercial feed (2% of biomass) and they were divided into two treatments: fish receiving food daily (control - fed), and fish submitted to cycles of 3 days of feed restriction and 2 days of re-feeding (no-fed group). For the in vitro experiments, oocytes (n = 20) were obtained from the ovaries removed at the end of the in vivo experiment and were divided into four groups: fed -IGF-1; fed +IGF-1; no-fed -IGF-1 and no-fed +IGF-1. Fish under restriction had lower body weights, decreased plasma glucose, increased triglycerides levels, and their final maturation and mature oocyte were reduced and the atresic ones were in higher number. Moreover, IGF-1, in vitro, increased the percentage of mature oocytes in fed females and decreased the atresic ones. In no-fed females, IGF-1 increased the final maturation and mature oocytes and reduced the atresic ones. This study demonstrates the importance of the feeding management of female breeders of matrinxã during the vitellogenesis period.

Long-term organic selenium supplementation overcomes the trade-off between immune and antioxidant systems in pacu (Piaractus mesopotamicus).

Selenium (Se) is an essential nutrient for antioxidant defenses in fish because of its role in preventing immunosuppression caused by oxidative stress. In this study it was demonstrated the relation between the oxidative stress and immune status after a long Se supplementation period, as a result of the evaluation of immunological, hematological and antioxidant responses, as well as growth performance of pacu fed diets supplemented with different concentrations of organic selenium (0, 0.3, 0.6, 0.9, and 1.8 mg Se-yeast/kg, but the final analyzed selenium concentrations were 0.72, 0.94, 1.15, 1.57 and 2.51 mg/kg, respectively) for 65 days. Dietary Se supplementation at 1.15 mg Se-yeast/kg (analyzed value) restored the production of antioxidant enzymes (glutathione peroxidase (GPx) and glutathione S-transferase (GST)), and consequently allowed the increased of some immunological parameters (leukocyte respiratory burst activity and lysozyme activity), hematological parameters (red blood cell count (RBC), hematocrit (HTC), mean corpuscular volume (MCV), and white blood cell count (WBC)). Se supplementation in pacu diets at 1.15 mg Se-yeast/kg for 65 days improved immune response and antioxidant defenses, suggesting that oxidative stress impairs immune system response to prevent excessive reactive oxygen species in cells and indicating the occurrence of a physiological trade-off between immune and antioxidant systems. Higher Se levels, such as 1.57 mg Se-yeast/kg increased the leukocyte respiratory burst activity, the WBC and thrombocyte counts, the RBC and HTC, and the GST and GPx enzymes. However, 2.51 mg Se-yeast/kg decreased the lysozyme levels, the WBC and thrombocyte counts, the RBC, HTC and MCV, and the GST and GPx enzymes. Those findings are important to future studies because showed the negative effect of oxidative stress on immunity, and may help to prevent any inhibition of the expected immune response after immunomodulators administration and vaccination. Also it was possible to meet the dietary selenium requirement of pacu, that was estimated to be 1.56 mg/kg.

Acute fluoxetine treatment increases aggressiveness in juvenile matrinxã (Brycon amazonicus).

Fluoxetine (FLX) is a selective serotonin (5-HT) reuptake inhibitor known for its effects modifying aggressiveness, personality traits, and anxiety-like behaviors. The aim of the present study was to evaluate the influence of the acute treatment, by immersion, with FLX on aggressive behavior of resident Brycon amazonicus fish. Fish pretreated with FLX presented an increase in aggressiveness, evidenced by the increase on the number of bites and chases against the intruder and a decrease in latency for the first attack, when compared to control fish. Together with previous studies, these results show the complexity of the neural modulation of the aggressive behavior in fish by 5-HTergic system.

Social challenge increases cortisol and hypothalamic monoamine levels in matrinxã (Brycon amazonicus).

The neural circuitry for social behavior and aggression appears to be evolutionarily conserved across the vertebrate subphylum and involves a complex neural network that includes the hypothalamus as a key structure. In the present study, we evaluated the changes in monoamine levels in the hypothalamus and on serum cortisol and plasma glucose of resident matrinxã (Brycon amazonicus) submitted to a social challenge (introduction of an intruder in their territory). The fight promoted a significant increase in hypothalamic 5-HT, NA and DA levels and on the metabolites 5-HIAA and DOPAC, and decreased 5-HIAA/5-HT and DOPAC/DA ratios in resident fish. Furthermore, an increase in serum cortisol and plasma glucose was also observed after the fight. Resident fish presented a high aggressiveness even with increased 5-HT levels in the hypothalamus. The alteration in hypothalamic monoaminergic activity of matrinxã suggests that this diencephalic region is involved in aggression and stress modulation in fish; however, it does not exclude the participation of other brain areas not tested here.

Novelty of the arena impairs the cortisol-related increase in the aggression of matrinxã (Brycon amazonicus).

The dichotomic effect of a cortisol level rise in vertebrate behavior has been widely observed. Generally, a chronic increase of the hormone level inhibits aggression, while an acute rise increases aggression. However, in this study, we show that this increase in aggression through an acute rise of cortisol also depends on the context in which the agonistic interaction occurs in the tropical fish matrinxã, Brycon amazonicus. We combined two factors: the type of housing (resident or non-resident in the trial arena) and the level of cortisol at the beginning of the fight (normal level - control, or high level - hydrocortisone-treated fish). The cortisol treatment increased the aggressiveness in the resident fish, but this effect was not observed in the non-resident fish, which fought in an unknown arena. The novelty of the arena may have elicited an "alerted state" in the non-resident fish; in this situation the fight was not the priority, and the cortisol effect in aggression could be impaired by a conflict between motivational systems (fear and aggression). In our knowledge, in fish, the increase of aggression promoted by an acute rise in cortisol levels was always tested and observed in a resident context, and the inhibition of cortisol effect in the agonist behavior is demonstrated for the first time. As the cortisol effect in aggression is observed in several taxa, the inhibition of aggressiveness increased by the novelty of the arena should be investigated in other groups to clarify the dynamics of this effect of cortisol in animal behavior.

Cortisol influences the antipredator behavior induced by chemical alarm cues in the Frillfin goby.

We evaluated the effect of increased plasma cortisol levels on fish antipredator behavior induced by conspecific chemical alarm cues. The experimental model for the study was the Frillfin goby Bathygobius soporator. We first confirmed that the alarm substance induces typical defensive antipredator responses in Frillfin gobies and described their alarm substance cells (epidermal 'club' cells). Second, we confirmed that intraperitoneal cortisol implants increase plasma cortisol levels in this species. We then demonstrated that exogenous cortisol administration and subsequent exposure to an alarm substance decreased swimming activity to a greater extent than the activity prompted by either stimulus alone. In addition, cortisol did not abolish the sheltering response to the alarm chemical cue even though it decreased activity. As predators use prey movements to guide their first contact with the prey, a factor that decreases swimming activity clearly increases the probability of survival. Consequently, this observation indicates that cortisol helps improve the antipredator response in fish.

Dietary L-tryptophan alters aggression in juvenile matrinxã Brycon amazonicus.

This study evaluated the effect of dietary supplementation with L: -tryptophan (L-TRP), a serotonin precursor, on the aggressiveness of juvenile matrinxã Brycon amazonicus. Fish were kept in individual aquaria for 7 days receiving the diets: D1 (control: 0.47% of TRP), D2 (0.94% of TRP), D3 (1.88% of TRP), and D4 (3.76% of TRP). After this, they were grouped with an intruder fish to establish a resident-intruder relationship during periods of 20 min. Blood cortisol, glucose, chloride, sodium and calcium; hemoglobin, hematocrit, red blood cell count and volume; liver glycogen and lipids were measured. Territoriality had significant effect on the aggressiveness of matrinxã (the residents were more aggressive than intruders, P < 0.001) and tryptophan significantly affected their behavior. Fish fed with the D2 diet presented a longer latency until the first attack (P = 0.0069) and bit the intruder fewer times (P = 0.0136) during the period of observation, compared to the control group. The frequency of bites and chases after the first attack was not affected by the dietary supplementation of TRP. Physiological variables were not significantly affected by the diet, except for a moderate increase in cortisol level in fish fed with D2 diet after the fight, indicating slight activation of the hypothalamus-pituitary-interrenal axis. The results show that juvenile matrinxã have aggressive and territorial behavior and that a diet containing 9.4 g TRP kg(-1) alter their aggressiveness, without affecting the stress-related physiological parameters.

Testicular structure and spermatogenesis of Amazonian freshwater cururu stingray Potamotrygon cf. histrix.

The cururu stingray Potamotrygon cf. histrix, a new and endemic Amazonian freshwater species, presents appropriate characteristics for fish keeping and is exploited from its natural environment. The present study identified the testicular structure and spermatogenesis of this species. Gonads from adult male specimens were dissected, fixed and processed for histological analysis. The testes were of testicular/epigonial type. The presence of germinal papillae was observed in the upper portion of organ with primordial germ cells and Sertoli cell precursors. The testis was lobular with zonal organization and cystic gametogenesis, with the occurrence of spermatoblasts. The Sertoli cells underwent morphological modifications over the course of gamete formation. The spermatozoids had long heads and were spiraled on their own axis. Information on the reproductive biology will serve as basis for studies on the reproduction and phylogeny of this peculiar group of cartilaginous fish.

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.

Desenvolvimento gonadal de fêmeas de matrinxã, Brycon amazonicus, submetidas a restrição alimentar / Gonadal development of matrinxã, Brycon amazonicus, females submitted to feed restriction

O presente estudo avaliou o efeito de ciclos de restrição alimentar e realimentação (2/3 dias), aplicados durante seis meses antes da desova, no desenvolvimento gonadal de matrinxã. Na ocasião da desova, fêmeas alimentadas diariamente e submetidas ao regime alimentar experimental, selecionadas para a indução hormonal, foram sacrificadas para retirada das gônadas e do fígado, com os quais se calculou o IGS (índice gonadossomático) e o IHS (índice hepatossomático), sendo os ovários processados para análise histológica. Não houve alteração no peso relativo dos ovários e fígado, e o desenvolvimento gonadal não foi afetado pelo esquema alimentar. Os valores de IGS foram de 5,09±4,98 por cento e 9,79±4,17 por cento e os de IHS foram de 0,84±0,07 por cento e 0,91±0,11 por cento, para as fêmeas controle e experimentais, respectivamente, sem diferenças significativas entre os grupos. Os ovários de peixes dos dois grupos apresentaram as mesmas características do estádio maduro, com predominância de ovócitos na fase final de maturação, repletos de vitelo. O estudo indica que a restrição alimentar não afetou a preparação das fêmeas para a reprodução e que ciclos adequados de restrição e realimentação poderão ser aplicados na criação do matrinxã, assegurando menores custos de produção.

The present study evaluated the effect of cycles of feed restriction and refeeding (2/3 days) during 6 months before the spawning on the gonadal development of matrinxã. At the spawning time, females selected to hormonal induction were killed for gonad and liver removal and GSI (gonadosomatic index) and HSI (hepatosomatic index) were calculated. Ovaries were processed for histological analysis. The feeding regime did not alter GSI and HSI and the gonadal development. The values of GSI were 5.09±4.98 percent and 9.79±4.17 percent, and of HSI were 0.84±0.07 percent and 0.91±0.11 percent, in control and experimental females, respectively, without significant difference between groups. Ovaries from both fish groups showed the same charachteristics of the mature stage, with predominance of oocytes in the final phase of maturation, presenting large amount of yolk. The study indicates that the feed restriction did not affect the matrinxã ability to reproduction and that cycles of feed restriction and refeeding can be used in matrinxã farming leading to lower production costs.

Influence of diets supplemented with vitamins C and E on pirarucu (Arapaima gigas) blood parameters.

This study evaluated the influence of diets supplemented with 500, 800, 1200 mg kg-1 of vitamin C (ascorbic acid or AA) and vitamin E (alpha-tocopherol or alpha-T) on the physiological responses of pirarucu fed for 2 months. Weight and mortality were not affected by dietary vitamin type or their concentrations. Significant increase (p<0.05) on the red blood cells count was obtained on treatments with 800 and 1200 mg AA kg-1 and on the hemoglobin concentration on treatment with 500 mg alpha-T kg-1 relatively to control. Mean corpuscular volume presented a significant decrease (p<0.05) on treatment with 800 and 1200 mg AA kg-1 when compared to control. Mean corpuscular hemoglobin concentration was significantly high (p<0.05) on treatment with 500 mg alpha-T kg-1. Only in vitamin C treatments, we noticed a significant increase (p<0.05) in the number of leucocytes relative to control. All fish in the vitamin-supplemented treatments, except 500 mg AA kg-1, had high total protein values compared to control. Fish treated with 800 or 1200 mg alpha-T kg-1 also showed increases in plasma glucose concentrations. Our results suggest that 800 and 1200 mg AA kg-1 are probably the most suitable concentrations for pirarucu diets, although high vitamin E diets are not necessary for quantitative leucocyte increases for this species.