Gene family evolution underlies cell-type diversification in the hypothalamus of teleosts.

Hundreds of cell types form the vertebrate brain but it is largely unknown how similar cellular repertoires are between or within species or how cell-type diversity evolves. To examine cell-type diversity across and within species, we performed single-cell RNA sequencing of ~130,000 hypothalamic cells from zebrafish (Danio rerio) and surface and cave morphs of Mexican tetra (Astyanax mexicanus). We found that over 75% of cell types were shared between zebrafish and Mexican tetra, which diverged from a common ancestor over 150 million years ago. Shared cell types displayed shifts in paralogue expression that were generated by subfunctionalization after genome duplication. Expression of terminal effector genes, such as neuropeptides, was more conserved than the expression of their associated transcriptional regulators. Species-specific cell types were enriched for the expression of species-specific genes and characterized by the neofunctionalization of expression patterns of members of recently expanded or contracted gene families. Comparisons between surface and cave morphs revealed differences in immune repertoires and transcriptional changes in neuropeptidergic cell types associated with genomic differences. The single-cell atlases presented here are a powerful resource to explore hypothalamic cell types and reveal how gene family evolution and shifts in paralogue expression contribute to cellular diversity.

Effects of vitamin A supplementation on ovarian development of Astyanax lacustris (Teleostei: Characidae) during the non-breeding season.

Vitamin A (VA) is essential for fish reproduction, however, knowledge of VA requirement of broodstock fish is scarce. We investigated the physiological role of VA supplementation in the ovarian maturation of Astyanax lacustris throughout the non-breeding season. Adult females were distributed in five nutritional treatments, which differed by the amount of VA supplemented in a plant-based diet: T1 = 0, T2 = 1800, T3 = 3600, T4 = 7200, and T5 = 14,400 IU kg-1 of VA for 60 days after a period of 16 days feeding on a diet poor in VA to reduce body VA content. Plasma total lipids (TL), lipoproteins, and 17ß-estradiol (E2) levels were measured after 30 and 60 days. Somatic indexes and morphophysiological ovarian parameters were calculated and VA concentration was analyzed in the ovaries and liver after 60 days. VA supplementation did not increase the concentration of this nutrient in the liver and ovaries. E2 plasma level increased in animals fed on 3600 IU kg-1 of VA (T3) after 60 days and females from T5 group presented a higher RF after 60 days. Ovarian development increased after 60 days compared to that after 30 days, confirmed by the higher gonadosomatic index (all groups). The supplementation with the highest VA level increased RF and decreased plasma TL (after 30 days) and low-density lipoprotein (LDL) (after 60 days) concentrations, showing a potential to improve reproduction even during the non-breeding season; however, its nutritional requirements of VA could be higher.

Dietary supplementation with nerolidol improves the antioxidant capacity and muscle fatty acid profile of Brycon amazonicus exposed to acute heat stress.

An increase in water temperature in the Amazon River has elicited concerns about commercially important fish species associated with food security, such as matrinxã (Brycon amazonicus). Studies have demonstrated the positive effects of diets supplemented with plant-based products that combat heat stress-induced oxidative damage. The aim of this study was to determine whether dietary supplementation with nerolidol prevents or reduces muscle oxidative damage and impairment of the fillet fatty acid profile of matrinxã exposed to heat stress. Plasma and muscle reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while plasma superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity was significantly lower. The total content of saturated fatty acids (SFA) in fillets was significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while he total content of polyunsaturated fatty acids (PUFA) was significantly lower. Nerolidol prevented the increase of muscle LPO and plasma ROS and LPO levels in fish exposed to heat stress, and partially prevented the increase in muscle ROS levels. Diets containing nerolidol prevented the inhibition of muscle GPx activity in fish exposed to heat stress, and partially prevented the decrease of plasma GPx activity. The nerolidol-supplemented diet prevented the increase of fillet SFA in fish exposed to heat stress, while partially preventing the decrease of PUFA. We conclude that acute heat stress at 34 °C for 72 h causes plasma and muscular oxidative damage, and that homeoviscous adaptation to maintain membrane fluidity can represent a negative impact for fish consumers. A nerolidol diet can be considered a strategy to prevent heat stress-induced oxidative damage and impairment of muscle fatty acid profiles.

Cavefish brain atlases reveal functional and anatomical convergence across independently evolved populations.

Environmental perturbation can drive behavioral evolution and associated changes in brain structure and function. The Mexican fish species, Astyanax mexicanus, includes eyed river-dwelling surface populations and multiple independently evolved populations of blind cavefish. We used whole-brain imaging and neuronal mapping of 684 larval fish to generate neuroanatomical atlases of surface fish and three different cave populations. Analyses of brain region volume and neural circuits associated with cavefish behavior identified evolutionary convergence in hindbrain and hypothalamic expansion, and changes in neurotransmitter systems, including increased numbers of catecholamine and hypocretin/orexin neurons. To define evolutionary changes in brain function, we performed whole-brain activity mapping associated with behavior. Hunting behavior evoked activity in sensory processing centers, while sleep-associated activity differed in the rostral zone of the hypothalamus and tegmentum. These atlases represent a comparative brain-wide study of intraspecies variation in vertebrates and provide a resource for studying the neural basis of behavioral evolution.

Evolution of the acoustic startle response of Mexican cavefish.

The ability to detect threatening stimuli and initiate an escape response is essential for survival and under stringent evolutionary pressure. In diverse fish species, acoustic stimuli activate Mauthner neurons, which initiate a C-start escape response. This reflexive behavior is highly conserved across aquatic species and provides a model for investigating the neural mechanism underlying the evolution of escape behavior. Here, we characterize evolved differences in the C-start response between populations of the Mexican cavefish, Astyanax mexicanus. Cave populations of A. mexicanus inhabit an environment devoid of light and macroscopic predators, resulting in evolved differences in various morphological and behavioral traits. We find that the C-start is present in river-dwelling surface fish and multiple populations of cavefish, but that response kinematics and probability differ between populations. The Pachón population of cavefish exhibits an increased response probability, a slower response latency and speed, and reduction of the maximum bend angle, revealing evolved differences between surface and cave populations. Analysis of the responses of two other independently evolved populations of cavefish, revealed the repeated evolution of reduced angular speed. Investigation of surface-cave hybrids reveals a correlation between angular speed and peak angle, suggesting these two kinematic characteristics are related at the genetic or functional levels. Together, these findings provide support for the use of A. mexicanus as a model to investigate the evolution of escape behavior.

Molecular characterization of different preproGnRHs in Astyanax altiparanae (Characiformes): Effects of GnRH on female reproduction.

Gonadotropin-releasing hormone (GnRH) is a key molecule in the initiation of the hypothalamic-pituitary-gonadal axis. Thus, knowledge about GnRH may contribute to the effectiveness of species reproduction. Using a Neotropical tetra Astyanax altiparanae as a fish model species, the GnRH forms were characterized at the molecular level and the role of injected GnRHs in vivo was evaluated. The full-length complementary DNA (cDNA) sequences of preproGnRH2 (612 bp) and preproGnRH3 (407 bp) of A. altiparanae were obtained, and the GnRH1 form was not detected. The cDNA sequences of preproGnRH2 and preproGnRH3 were found to be conserved, but a change in the amino acid at position 8 of the GnRH3 decapeptide of A. altiparanae was observed. All the injected GnRHs stimulated lhß messenger RNA (mRNA) expression but not fshß mRNA expression, and only GnRH2 was able to increase maturation-inducing steroid (MIS) levels and possibly stimulate oocyte release. Furthermore, only GnRH2 was able to start the entire reproductive hormonal cascade and induce spawning.

From collection to discharge: physical, chemical, and biological analyses for fish farm water quality monitoring.

The use of chemical substances for the management of fish farming activities may compromise the quality of the tank water itself and of water bodies that receive the effluents. As studies that assess the environmental effect caused by pisciculture are scarce, the present study aimed at evaluating the water quality in two fish farms in the region of Grande Dourados, Brazil, from the site of water collection to the site of water disposal. The tools used for this purpose were the analysis of land use and cover and the determination of physical, chemical, and biological parameters of water samples. Maps of land use and cover were created, and water samples were collected at four sampling sites in two fish farms. The Allium cepa test, assays with Astyanax lacustris, and the Salmonella/microsome assay were performed. In addition, physical and chemical parameters were measured and metal and emerging contaminants in the water samples were investigated. The A. lacustris demonstrated the genotoxicity and the Salmonella/microsome assay suggested the mutagenic potential of water samples from the fish farms and indicated higher genotoxicity in the disposal tanks than in the collection tanks of the Brilhante fish farm. However, all the samples at the Dourados fish farm were genotoxic, and mutagenicity was shown to start at the water collection site. With regard to the A. cepa test, there was no statistical difference between the collection sites in both fish farms. Moreover, the observed genetic damage may be associated with the presence of metals and emerging contaminants in the water samples, which suggests that these chemicals have potential genotoxic and mutagenic effects that are related to the type of land use and cover in the area of the region studied. Considering that contaminated waters can potentially disturb the structure and functioning of natural ecosystems, the present study demonstrated the importance of treating fish farm effluent to minimize the negative effect of this activity on water bodies.

Hypocretin underlies the evolution of sleep loss in the Mexican cavefish.

The duration of sleep varies dramatically between species, yet little is known about the genetic basis or evolutionary factors driving this variation in behavior. The Mexican cavefish, Astyanax mexicanus, exists as surface populations that inhabit rivers, and multiple cave populations with convergent evolution on sleep loss. The number of Hypocretin/Orexin (HCRT)-positive hypothalamic neurons is increased significantly in cavefish, and HCRT is upregulated at both the transcript and protein levels. Pharmacological or genetic inhibition of HCRT signaling increases sleep in cavefish, suggesting enhanced HCRT signaling underlies the evolution of sleep loss. Ablation of the lateral line or starvation, manipulations that selectively promote sleep in cavefish, inhibit hcrt expression in cavefish while having little effect on surface fish. These findings provide the first evidence of genetic and neuronal changes that contribute to the evolution of sleep loss, and support a conserved role for HCRT in sleep regulation.

Aloe vera enhances the innate immune response of pacu (Piaractus mesopotamicus) after transport stress and combined heat killed Aeromonas hydrophila infection.

In this study, pacu (Piaractus mesopotamicus) were fed with diets containing Aloe vera for 10 days prior to transport stress and infection with heat killed Aeromonas hydrophila. A. vera is popular around the world due to its medicinal properties, including immunostimulatory effects which was observed in this study. The results show that transport causes immunosuppression, an effect that was prevented by A. vera. Specifically, A. vera prevented reductions of both leukocyte respiratory burst and hemolytic activity of complement system caused by transport. Further, fish fed with A. vera also showed significantly higher leukocyte respiratory burst, serum lysozyme concentrations and activity of complement system 24 h after bacterial infection. Additionally, we observed that A. vera may modulate the innate response through activation of complement system during bacterial immune stimulation. In summary, A. vera extract enhanced innate immune parameters and consequently the ability of fish to cope with pathogens following transport stress. These findings show that A. vera has promise for use in aquaculture and add further evidence that medicinal herbs added to fish feed assist to prevent disease outbreaks.

Is There Volume Transmission Along Extracellular Fluid Pathways Corresponding to the Acupuncture Meridians?

Volume transmission is a new major communication signaling via extracellular fluid (interstitial fluid) pathways. It was proposed by the current authors that such pathways can explain the meridian phenomena and acupuncture effects. To investigate whether meridian-like structures exist in fish body and operate via volume transmission in extracellular fluid pathways, we injected alcian blue (AB) under anesthesia into Gephyrocharax melanocheir, which has a translucent body. The migration of AB could be seen directly and was recorded by a digital camera. The fish was then embedded and cut transversely to observe the position of tracks in three dimensions. Eight longitudinal threadlike blue tracks were recognized on the fish. The positions of these threadlike tracks were similar to meridians on the human body. Transverse sections showed that these tracks distributed to different layers of distinct subcutaneous loose connective tissues and intermuscular septa. Lymphatic vessels were sometimes associated with the extracellular blue tracks where the migration of AB occurred. Extracellular fluid pathways were found on fish through their transport of AB. These pathways operating via volume transmission appeared to be similar in positions and functions to the acupuncture meridians in Chinese medicine.

Curcuma longa as additive in the diet for Astyanax aff. bimaculatus.

The aim of the present study was to evaluate the effect of turmeric (Curcuma longa) as additive in the diet for Astyanax aff. bimaculatus. Fish (0.83 ± 0.04 g) were fed, for 60 days, with six diets containing 0.0, 20.0, 40.0, 60.0, 80.0, and 100.0 g turmeric kg-1 feed. There was an increasing linear effect of turmeric on the thickness of the muscular layer, and height and width of the folds of the intestine. In the liver, a quadratic effect was observed of turmeric on the percentage of hepatocyte cytoplasm and a decreasing linear effect on the percentage of sinusoid capillaries. A quadratic effect was also observed of turmeric on the liver glycogen. There was no effect of turmeric on the antioxidant activity in the liver, carcass composition or productive performance of the fish. Thus, we concluded that Curcuma longa has trophic effects on the epithelium and the muscular layer of the intestine of A. aff. bimaculatus. Additionally, low levels of Curcuma longa cause increased deposition of liver glycogen and high levels cause reduction.

ß-Glucan-induced cortisol levels improve the early immune response in matrinxã (Brycon amazonicus).

This study investigated the role of endogenous cortisol on the innate immune response in matrinxã (Brycon amazonicus) fed with ß-glucan, prior to and after stressor exposure and bacterial challenge. For this, we evaluated the serum cortisol and plasma glucose levels, the serum lysozyme levels, the hemolytic activity of the complement system, and the respiratory activity of leukocytes, as well as the number of circulating erythrocytes and leukocytes of fish fed during 15 days with diets containing ß-glucan 0.1% (ß-G) or ß-glucan 0.1% + metyrapone 30 mg kg-1 fish (ß-G + MTP). Dietary MTP was used to block cortisol production. After feeding, fish were air-exposed during 3 min, to endogenously increase the cortisol levels. Following that, they were challenged with intraperitoneal injection of Aeromonas hydrophila. 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 the handling during injection. Fish were sampled prior to the stressor exposure, 30 min after exposure, and 24 h post infection (hpi). Herein we observed that dietary ß-G modulated the cortisol profile prior to and after the stressor, increasing the number and activity of leukocytes. Moreover, cortisol showed to be an efficient modulator of both humoral and cellular innate immune system by increasing lysozyme and complement activity, as well as neutrophil and monocyte populations. Our results suggest that ß-glucan-induced cortisol increase is one important mechanism to improve the innate immune response in matrinxã.

Wound healing in Piaractus mesopotamicus supplemented with chromium carbochelate and Saccharomyces cerevisiae / Cicatrização cutânea em Piaractus mesopotamicus suplementados com carboquelato de cromo e Saccharomyces cerevisiae

Wound healing and tissue repair are necessary to ensure survival and health of any organism. The aim of this study was to investigate the impact of supplementation with chromium carbochelate (CC) and Saccharomyces cerevisiae (SC) on wound healing in tropical teleost fish Piaractus mesopotamicus. Thus, fish were distributed into four groups: a) control (without supplementation); b) supplemented with 18 mg/kg of chromium carbochelate; c) supplemented with 0.3% of S. cerevisiae and d) supplemented with an association of both supplements. After 105 days of feeding, full-thickness skin incisions (2.0 x 1.0 x 0.25 cm) were performed removing epidermis and dermis. Macroscopic and histologic observations were carried out at 1, 3, 7, 14, 21, 28, and 35 days after wounding to monitor the healing rate. Opposing fronts advanced gradually and faster each day demonstrating a progressive increase in the healing process over time. The inflammatory process was exacerbated and expansive, with an increase in mucous cells and chromatophores. Although no significant differences were observed between groups on wound retraction and microscopic parameters, fish supplemented with CC and SC showed faster re-epithelialization, greater degree of organization of collagen fibers, and higher neovascularization. We concluded that supplementation with S. cerevisiae and chromium carbochelate improves specific aspects of cutaneous healing process in pacu.(AU)

A cicatrização e reparação de tecidos são mecanismos essenciais para garantir a sobrevivência e saúde de qualquer indivíduo. O objetivo deste trabalho foi avaliar o impacto da suplementação com carboquelato de cromo (CC) e Saccharomyces cerevisiae (SC) sobre a cicatrização no peixe tropical Piaractus mesopotamicus. Para isto, os peixes foram distribuídos em quatro grupos: controle (sem tratamento), suplementados com 18 mg/kg de carboquelato de cromo, 0,3% de S. cerevisiae e associação de ambos os suplementos. Após 105 dias de alimentação, foram realizadas incisões na pele de espessura completa (2,0 x 1,0 x 0,25 cm) removendo epiderme e derme. Avaliações macroscópicas e microscópicas foram realizadas 1, 3, 7, 14, 21, 28 e 35 dias após a indução das feridas, para monitorar a taxa de cicatrização. As bordas opostas das feridas avançaram gradualmente a cada dia, demonstrando o aumento progressivo do processo de cicatrização ao longo do tempo. O processo inflamatório foi exacerbado e expansivo, com aumento no número de células mucosas e cromatóforos. Apesar deste processo, não foram observadas diferenças significativas na retração das feridas e nos parâmetros microscópicos entre os grupos. Peixes suplementados com CC ou Sc apresentaram rápida reepitelização, maior grau de organização de fibras colágenas e de neovascularização inicial. Concluiu-se que a suplementação com S. cerevisiae ou carboquelato de cromo melhora aspectos específicos do processo cicatricial no pacu.(AU)

Appetite regulating factors in dourado, Salminus brasiliensis: cDNA cloning and effects of fasting and feeding on gene expression.

The dourado, Salminus brasiliensis (Cuvier, 1816) is a freshwater piscivorous Characin native to South American rivers. Owing to the high quality of its flesh and its fast growth, it is the object of both capture fisheries and fish farming. However, very little is known about the endocrine regulation of feeding and metabolism of dourado. In this study, cDNAs for orexin, CART and CCK were isolated in dourado, and their mRNA tissue distributions examined. In order to assess the role of these peptides in the regulation of feeding of dourado, the effects of fasting and feeding on mRNA expression levels of orexin, CART and CCK in the brain as well as CCK in the intestine were assessed. Whereas orexin and CCK have widespread mRNA distributions in the brain and peripheral organs, CART seems to be mostly limited to the brain. Orexin brain expression increased with fasting and displayed periprandial changes, suggesting it is involved in both long- and short-term regulation of feeding and appetite. CART and CCK hypothalamic expressions were not affected by fasting, but displayed periprandial changes with post-feeding decreases, suggesting roles in short-term satiation. CCK expression in the anterior intestine was not affected by fasting and did not display periprandial changes. Overall, our results suggest that orexin, CART and CCK are involved in the physiology of feeding of dourado.

Intestinal and liver morphometry of the Yellow Tail Tetra (Astyanax altiparanae) fed with oregano oil.

This study aimed to evaluate the effect of oregano oil on the intestinal and liver morphometry of yellow tail tetra, Astyanax altiparanae. Fish (1.46 ± 0.09 g) were kept in a 60-L aquaria, at a stocking density of 0.5 fi sh L-1. Six diets containing varying amounts of oregano oil were evaluated (0.0; 0.5; 1.0; 1.5; 2.0 and 2.5 g of oregano oil kg-1). At the end of 90 days, the fi sh were euthanised. Four intestines and four livers were collected per treatment, which were fi xed in Bouin and embedded in resin. For height and width folds, the absorption surface area and thickness of the muscular layer a positive linear effect of oregano oil was observed. A decrescent linear effect on the total number of goblet cells was also observed. For the cytoplasmic percentage of hepatocytes and liver glycogen, a positive linear effect of oregano oil was observed. There was a decreasing linear effect on the percentage of nuclei in the hepatocytes and capillaries. Thus, the oregano essential oil promotes increased absorption areas, modulates the amount of goblet cells involved in protecting the intestinal mucosa and promotes cytoplasmic increase with greater deposition of liver glycogen in yellow tail tetra.

Hematological responses of tambaqui Colossoma macropomum (Serrassalmidae) fed with diets supplemented with essential oil from Mentha piperita (Lamiaceae) and challenged with Aeromonas hydrophila / Respostas hematológicas de tambaqui Colossoma macropomum (Serrassalmidae) alimentado com dietas suplementadas com óleo essencial de Mentha piperita (Lamiaceae) e desafiados com Aeromonas hydrophila

In fish farmings, diseases can be reduced by using immunostimulants. The aim of this study was to evaluate the immunostimulant potential of Mentha piperita in tambaqui fed with 0, 0.5, 1.0 and 1.5% of oil per kg of commercial fish feed. The fish were inoculated with Aeromonas hydrophila to challenge them. Hematological and biochemical parameters were determined after 30 days of feeding and seven days after the challenge. There was no mortality and M. piperita oil did not influence fish production parameters. However, blood hemoglobin concentration (Hb) increased in the fish fed with 0.5 and 1.5% of oil per kg of diet; albumin increased in those fed with 1.0%; cholesterol increased in all groups with oil; and triglycerides increased in those fed with 0.5%. After the bacterial challenge, the fish showed decreases in Hb when fed with diet enriched with 1.5% oil per kg of diet, in mean corpuscular volume with 1.0% and in mean corpuscular hemoglobin concentration with 0 and 1.5%. Protein levels increased in groups with 0 and 1.5% of oil and albumin when fed with 0 and 1.0%; cholesterol levels increased in the control group; and high levels of triglycerides were observed in the groups with 0, 0.5 and 1.5%. Thus, M. piperita essential oil promoted hematological alterations in tambaqui and can be recommended in diets containing up to 1.0% per kg, because of the minimal physiological modifications caused. However, additional studies are necessary to obtain more information regarding to the physiological effects of this immunostimulant.

Em pisciculturas, doenças podem ser minimizadas com o uso de imunoestimulantes. O objetivo deste estudo foi avaliar o potencial imunoestimulante do óleo essencial de Mentha piperita na dieta de tambaqui com dietas contendo 0; 0,5; 1,0; 1,5% de inclusão do óleo por kg de ração comercial para peixes. Os tambaquis foram desafiados com a bactéria Aeromonas hydrophila. Hematologia e bioquímica dos tambaquis foram determinados após 30 dias de alimentação e sete dias do desafio. Não houve mortalidade dos peixes durante o experimento, sendo que o óleo não influenciou os parâmetros produtivos dos peixes. Entretanto, a concentração de hemoglobina (Hb) aumentou em peixes alimentados com 0,5 e 1,5% de óleo por kg de dieta; a albumina aumentou em 1,0%; o colesterol aumentou em todos os grupos com óleo; e triglicérides em 0,5% por kg de dieta. Após desafio bacteriano ocorreram diminuições da Hb nos peixes alimentados com 1,5% de óleo; do volume corpuscular médio com 1,0%; e da concentração de hemoglobina corpuscular média com 0 e 1,5%. Níveis aumentados de proteína nos grupos 0 e 1,5%; da albumina em 0 e 1,0%; do colesterol no controle; e dos triglicérides em 0, 0,5 e 1,5% de óleo na dieta. Assim o óleo essencial de M. piperita na dieta promoveu alterações hematológicas e pode ser recomendado seu uso em inclusões de até 1,0% por kg da dieta de tambaqui, devido às mínimas alterações fisiológicas causadas. Entretanto, estudos adicionais são necessários, de modo a obter informações sobre os efeitos fisiológicos deste imunoestimulante.

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.

Growth and intestinal morphology of juvenile pacu Piaractus mesopotamicus (Holmberg 1887) fed dietary prebiotics (mannanoligosaccharides - MOS).

Intensification of aquaculture production systems exposes fish to numerous stressors, which may negatively affect their growth and limit profitability of aquaculture systems. This study determined effects of increasing levels of dietary mannanoligosaccharides on growth and intestine morphology of pacu. Fish (44.04 g) were randomly distributed into 32 tanks (500 L; 10 fishes per tank) and fed during 63 days with a commercial diet supplemented with 0.0; 0.2; 0.4; 0.6; 0.8; 1.0; 1.5 and 2.0% dietary mannanoligosaccharides. Growth parameters did not differ (P>0.05) between fish fed control diet and mannanoligosaccharide supplemented diets. Intestinal villi perimeter was performed in fish fed control diet, 0.4 and 1.5% dietary mannanoligosaccharides and also showed no differences (P>0.05) between treatments. Dietary supplementation of mannanoligosaccharides unclear did not have effects on pacu. Studies on the characterization of intestinal microbiota together with experiment that reproduce commercial fish production systems rearing conditions are necessary to determine the effective use of this dietary supplement for the species.

The time course of aggressive behaviour in juvenile matrinxã Brycon amazonicus fed with dietary L-tryptophan supplementation.

This study evaluated the influence of dietary L-tryptophan (TRP) supplementation on the time course of aggressive behaviour and on neuroendocrine and hormonal indicators in juvenile matrinxã Brycon amazonicus. Supplementation with TRP promoted a change in the fight pattern at the beginning of an interaction with an intruder, resulting in decreased aggressive behaviours during the first 20 min. The decrease in aggression did not persist throughout the interaction but increased at 3 and 6 h after the beginning of the fight. Monoamine levels in the hypothalamus were not influenced by TRP before or after the fight; however, the hypothalamic serotonin (5-HT) concentration and the 5-hydroxyindole-3-acetic acid (5HIAA):5-HT ratio were significantly correlated with the reduction in aggressive behaviour at the beginning of the fight. Cortisol was not altered by TRP before the fight. After the fight cortisol increased to higher levels in B. amazonicus fed with supplementary TRP. These results indicate that TRP supplementation alters the aggressive behaviour of B. amazonicus and that this effect is limited to the beginning of the fight, suggesting a transient effect of TRP on aggressive behaviour. This is the first study reporting the effects of TRP supplementation on the time course of aggressive interaction in fishes.

Growth and hematology of juvenile pacu Piaractus mesopotamicus (Holmberg 1887) fed with increasing levels of vitamin E (DL-α-tocopheryl acetate).

Intensive fish production systems are characterized by 100% artificial feeding, so any dietary imbalances or deficiencies may lead to diseases outbreaks and economic losses. This study was set out to determine the effects of increasing levels of dietary vitamin E on growth and hematology of juvenile pacu. Fishes were fed for 90 days, twice a day until apparent satiation with semi-purified diets containing 0.0; 25; 50; 150; 300 or 600 mg.kg-1 diet DL-α-tocopheryl acetate in a completely randomized design trial (n=4); biometrical and hematological data were collected and analyzed. Fishes fed with vit E diet (150 mg.kg-1) showed higher (p<0.05) weight gain and specific growth. Hematocrit, erythroblast number and total plasma protein were increased (p<0.05) in fishes fed diet with no vit E diet. Vitamin E supplementation in artificial diets for pacu is essential for growth and maintenance of normal erythropoiesis.