Effect of digestible protein on intermediate metabolism, hepatic enzyme activities, energy reserves, and growth performance of pacu (Piaractus mesopotamicus) in the finishing growth phase.

This study investigated the effect of different levels of digestible protein (DP) on blood metabolites, hepatic enzyme activity of glycolysis and amino acid metabolism, energy reserves, and the production characteristics of pacu (Piaractus mesopotamicus) during the finishing growth phase. Six semi purified and isoenergetic diets, containing 16.3, 20.1, 23.8, 27.2, 31.5, and 34.8% of balanced DP, provided with essential amino acid balance, were hand-fed to pacu (1100.0 ± 10.3 g, initial weight) three times daily for 7 weeks. The experiment consisted of six treatments, with three randomly arranged replicates (tanks) per treatment. The data obtained from this experiment were analyzed by one-way analysis of variance (ANOVA), and significant differences (p < 0.05) between treatments were determined using Tukey's test. Blood metabolites, except serum ammonia and the hepatic enzymes activities of glycolysis and amino acid metabolism, except hexokinase activity were affected (p < 0.05) by balanced DP. The energy reserve indices, except hepatic total lipid content, were also found associated (p < 0.05) with balanced DP. The test diets significantly (p < 0.05) affected growth performance parameters. Higher dietary proteins led to a greater energy uptake by fish from the protein in feed. Overall, fish fed the intermediate level (23.8%) of balanced DP with digestible energy of 17.95 MJ kg-1 showed better production traits and physio-biochemical health markers. This information could help nutritionists and farmers to develop nutritionally balanced and economically and environmentally sustainable aquafeed for promoting healthy and sustainable production of pacu in intensive culture systems.

First Report of Aeromonas veronii as an Emerging Bacterial Pathogen of Farmed Nile Tilapia (Oreochromis niloticus) in Brazil.

Brazil is one of the world's leading producers of Nile tilapia, Oreochromis niloticus. However, the industry faces a major challenge in terms of infectious diseases, as at least five new pathogens have been formally described in the last five years. Aeromonas species are Gram-negative anaerobic bacteria that are often described as fish pathogens causing Motile Aeromonas Septicemia (MAS). In late December 2022, an epidemic outbreak was reported in farmed Nile tilapia in the state of São Paulo, Brazil, characterized by clinical signs and gross pathology suggestive of MAS. The objective of this study was to isolate, identify, and characterize in vitro and in vivo the causative agent of this epidemic outbreak. The bacterial isolates were identified as Aeromonas veronii based on the homology of 16S rRNA (99.9%), gyrB (98.9%), and the rpoB gene (99.1%). A. veronii showed susceptibility only to florfenicol, while it was resistant to the other three antimicrobials tested, oxytetracycline, enrofloxacin, and amoxicillin. The lowest florfenicol concentration capable of inhibiting bacterial growth was ≤0.5 µg/mL. The phenotypic resistance of the A. veronii isolate observed for quinolones and tetracycline was genetically confirmed by the presence of the qnrS2 (colE plasmid) and tetA antibiotic-resistant genes, respectively. A. veronii isolate was highly pathogenic in juvenile Nile tilapia tested in vivo, showing a mortality rate ranging from 3 to 100% in the lowest (1.2 × 104) and highest (1.2 × 108) bacterial dose groups, respectively. To our knowledge, this study would constitute the first report of highly pathogenic and multidrug-resistant A. veronii associated with outbreaks and high mortality rates in tilapia farmed in commercial net cages in Brazil.

Carbohydrate tolerance in Amazon tambaqui (Colossoma macropomum) revealed by NMR-metabolomics - Are glucose and fructose different sugars for fruit-eating fish?

In the present study, two approaches were followed to evaluate the metabolic responses of tambaqui (Colossoma macropomum), a frugivorous species, to intraperitoneal (IP) administration of glucose (GLU) and fructose (FRU) in fed (FED) and 10-day fasted (FAST) fish. Glucose and fructose tolerance tests were performed to assess the carbohydrate utilization and complementary NMR-metabolomics analyses were done to elucidate the impacts of sugar mobilization on the metabolic profile of plasma, liver and muscle. Blood was sampled from FED groups at 0, 3, 6 and 24 h; and at 0 and 24 h from FAST groups. Significant differences were observed in the hyperglycaemic peak between sugars at 3 h (GLU - 13.7 ± 2.0 mM vs. FRU - 8.7 ± 1.1 mM; saline 6.3 ± 0.6 mM) and on the return to normoglycaemia (GLU - 8.5 ± 2.2 mM vs. FRU - 5.2 ± 0.9 mM; saline 4.9 ± 0.6 mM) 6 h after IP on the FRU fish. The NMR-metabolomics approach allowed to conclude that tambaqui seems to be more responsive to the feeding regime (FED vs. FAST) than to the injected sugar (FRU vs. GLU). From the studied tissues, plasma showed no significant variations between feeding regimes at 24 h after IP, while muscle and liver revealed some variations on the final metabolome profile between FED and FAST groups. The metabolome variations between feeding regimes are indicative of changes on the amino acid utilization. Fish from FAST group seem to utilize amino acids as energy source rather than for protein synthesis and muscle growth. Variations on glucose concentration in muscle can also indicate different utilization of the sugars depending on the feeding regime.

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.

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.

Estratégia alimentar com ciclos de restrição e realimentação no desempenho produtivo de juvenis de tilápia do Nilo da linhagem GIFT / Feeding strategies with cycles of restriction and refeeding on growth performance of Nile tilapia juveniles, GIFT line

O objetivo deste estudo foi avaliar a ocorrência de resposta compensatória no desempenho produtivo de juvenis de tilápia do Nilo Oreochromis niloticus, linhagem GIFT, submetidos a diferentes estratégias alimentares. Foram utilizados 135 juvenis de tilápia, distribuídos em nove tanques de polietileno de 100L cada. As estratégias testadas foram: grupo controle (alimentado todo dia), grupo alimentado por cinco dias seguidos de dois dias de restrição de alimento (5A/2R) e grupo alimentado por quatro dias seguidos de três dias de restrição de alimento (4A/3R). Foram avaliados parâmetros físico-químicos da água e de desempenho produtivo. Os resultados foram submetidos à análise de variância, e as médias foram comparadas pelo teste Tukey, a 5 por cento de probabilidade. A qualidade de água, o fator de condição e a conversão alimentar não foram influenciados pela estratégia alimentar. O grupo alimentado com a estratégia 5A/2R apresentou peso final, ganho de peso e taxa de crescimento específico semelhantes ao grupo continuamente alimentado (7,8 e 9,2g; 6,4 e 7,8g e 2,7 e 3,0 por cento dia-1, para peso final, ganho de peso e taxa de crescimento especifico, respectivamente). A estratégia 4A/3R apresentou os piores resultados de desempenho produtivo, e a estratégia 5A/2R pode ser usada na alimentação de juvenis de tilápia do Nilo sem prejuízo ao desempenho produtivo, possibilitando inclusive redução de até 22,5 por cento na quantidade de alimento ofertada.

The objective of this study was to evaluate the occurrence of compensatory answer on productive performance of Nile tilapia Oreochromis niloticus juveniles, GIFT line, submitted to different feeding strategies. It was used 135 juveniles tilapias distributed in nine 100L polyethylene tanks each. The tested strategies were the control group (fed everyday), group fed for 5 days followed by 2 days of food restriction (5A/2R) and group fed for 4 days followed by 3 days of food restriction (4A/3R). Were evaluated physical chemical water parameters e growth performance. Results were submitted to an analysis of variance and averages compared by Tukey test 5 percent. Water quality, condition factor and feed conversion were not influenced by feeding strategy. Group fed 5A/2R strategy showed final weight, weight gain and specific growth rate similar to group continually fed (7.8 e 9.2g; 6.4 e 7.8g e 2.7 e 3.0 percent day-1, for final weight, weight gain and specific growth rate, respectively). The strategy 4A/3R showed the poorest productive performance. Strategy 5A/2R can be used in Nile tilapia juveniles feeding without damage to the fish productive performance, allowing a reduction of up to 22.5 percent on the feed quantity offered.