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The aim of this research was to assess the effects of different feeding regimes on growth performance, plasma immune indices and related genes' expression of hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) juveniles fed two dietary energy levels. A 3 × 2 factorial experiment was designed including three feeding regimes (two meals a day, one meal a day, one meal every two days; designated as 2 M, 1 M, 2DM) and two dietary available energy levels (3.4 and 3.6 kcal/g; designated as E3.4, E3.6), with three replicates per treatment. Juvenile hybrid grouper (average initial body weight: 19.39 ± 0.04 g/fish) were cultured for 6 weeks in 18 floating cages (125 cm × 80 cm × 40 cm) placed in six concrete ponds sharing the same water source. Results revealed that the 2 M and 1 M treatments had significantly higher weight gain rate (WGR), protein efficiency ratio (PER), expression of hepatic IGF-1, TOR, and S6K1 than the 2DM treatment (P < 0.05), whereas feed conversion ratio (FCR) displayed an opposite trend (P < 0.05). No significant differences in the above parameters were found between the 2 M and 1 M groups. As the feeding frequency decreased, the total feed intake (FI) per fish was significantly reduced (P < 0.05), while feed intake per meal (FIM), stomachic index (SI) and expression of hypothalamus agouti related neuropeptide (AgRP) were significantly increased (P < 0.05). The 2 M treatment supported higher plasma lysozyme contents than the 1 M and 2DM treatments (P < 0.05). The E3.6 treatment had higher intraperitoneal fat ratio and intestinosomatic index, but lower plasma lysozyme and immunoglobulin M contents than the E3.4 treatment (P < 0.05). Based on our results, a feeding regime of two meals/day of a diet containing 3.4 kcal DE/g of dry matter is recommended for the culture of juvenile hybrid grouper with an initial size of 19.39 g. • The feeding regime of two meals a day is suitable for hybrid grouper. • The feeding regime of one meal every two days impairs fish growth and health. • The feeding regimes affect expression of genes involved in the TOR/S6K1 pathway. • The high dietary energy level (358 kcal per 100 g) impairs immune indices of hybrid grouper. [ABSTRACT FROM AUTHOR]

The optimum phenylalanine (Phe) requirement for hybrid grouper (Epinephelusfuscoguttatus ♀ × Epinepheluslanceolatus ♂) juveniles was determined through an 8-week growth trial. A total of seven isoenergetic (340 kcal per 100 g of dry matter), isonitrogenous, and isolipidic diets were made, containing 8.2 (Phe 8.2), 9.2 (Phe 9.2), 10.1 (Phe 10.1), 11.2 (Phe 11.2), 13.3 (Phe 13.3), 15.2 (Phe 15.2), and 17.3 g/kg (Phe 17.3), respectively. Triplicate tanks of juvenile fish (about 16.7 g/fish) were fed each experimental diet twice daily until apparent satiation. The results indicated that different dietary Phe levels significantly influenced weight gain percentage (WG), feed efficiency (FE), protein efficiency ratio (PER), as well as, productive protein value (PPV). Fish fed Phe 8.2 had the lowest WG or PPV among all experimental treatments. Furthermore, the optimal dietary Phe level increased fold height, width, enterocyte, and microvillus height of fish. The Phe 10.1 group exhibited higher growth hormone (GH) expression in the pituitary compared to other groups. Expression of hepatic insulin-like growth factor-1 (IGF-1) and growth hormone receptor 1 (GHR1) displayed a similar pattern of variation to that of GH. The Phe 13.3 group had lower expression of S6 kinase 1 (S6K1) and target of rapamycin (TOR) than other groups. In addition, fish fed Phe 10.1 had lower levels of nuclear factor erythroid 2 (Nrf2) and heat shock protein 70 (HSP70) in the head kidney, and Cu/Zn-superoxide (Cu/ZnSOD) dismutases in the midgut compared to fish fed other Phe levels. Generally, optimal Phe content in the diet of hybrid grouper was estimated to be 12.7 g/kg of dry matter (27.3 g/kg of dietary protein), and at this level, the feed utilization, gut micromorphology, and immunity of fish were also elevated. [ABSTRACT FROM AUTHOR]

A six-week feeding trial was carried out to evaluate the effects of different dietary Myo -inositol (MI) levels on growth, feed efficiency, immunity, and antioxidant properties of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) fed low-fishmeal diets. Seven experimental diets were made to contain 50, 100, 169, 241, 309, 386, and 459 mg/kg of MI (analyzed values, dry matter basis), being abbreviated as MI-50, MI-100, MI-169, MI-241, MI-309, MI-386, and MI-459, and each MI treatment had three replicates. Twelve fish (average initial body weight: 18.7 ± 0.1 g) were allocated to each replicate, being hand-fed their diets to apparent satiation on 8:00 am and 16:30 pm. The results indicated that within the range of 50–241 mg/kg dietary inositol, weight gain percentage (WG%), feed efficiency (FE) and protein efficiency ratio (PER) were increased with the increasing of MI, and above the 241 mg/kg, they were dropped, especially at 386 and 459 mg/kg. According to the broken-line regression analysis conducted to assess the relationship between WG% and dietary MI levels, the optimum MI dose was 267.9 mg/kg (dry weight) in the low-fishmeal diet of hybrid grouper. Condition factor (CF), viscerosomatic index (VSI), hepatosomatic index (HSI), and intestinal index (ISI) were significantly (P < 0.05) affected by dietary MI levels, varying with a similar trend as WG%. Fish fed MI-241 had the highest values of serum lysozyme (LZM), hepatic total antioxidant capacity (T-AOC) as well as the relative mRNA level of hepatic insulin-like growth factor-Ⅰ (IGF-Ⅰ) among all treatments. In conclusion, optimum dietary MI requirement for the maximal growth of hybrid grouper was 267.9 mg/kg, and suitable dietary MI supplementations improve growth, immunity, and antioxidant properties of juvenile hybrid grouper fed low-fishmeal diets. • An optimal MI content of 267.9 mg/kg is recommended in the low-fishmeal diets of hybrid grouper. • Suitable MI supplementations improve growth, immunity and antioxidant properties of fish. • Dietary MI levels exceeded 368 mg/kg have a negative impact on fish. [ABSTRACT FROM AUTHOR]

Ascorbic acid (Vitamin C - VC) is one of the essential micronutrients required for normal growth and physiology of fish. In this research, the optimum dietary VC requirement of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) was evaluated in a comparative feeding trial and subsequent oxidative stress test. Six isoenergetic (340 kcal/kg), isoproteic (49%) and isolipidic (7%) experimental diets were supplemented with graded amounts of L-ascorbyl-2-polyphosphate (analyzed VC concentrations of 76, 86, 104, 126, 137 and 152 mg/kg dry matter) and hand-fed to fish (initial weight of 15 ± 0.1 g/fish (means ± S.D.)) in three replicate cages twice a day (08:00 h and 16:30 h) to apparent satiation for 7 weeks. Percent weight gain (WG%), together with the VC concentrations in liver and head kidney, were elevated with increasing dietary VC contents, reaching a plateau in the VC137 treatment. The optimal dietary VC requirement of hybrid grouper was estimated to be 148 mg/kg dry matter based on broken-line regression analysis of WG%. Serum immunoglobulin M (IgM) concentrations and the relative mRNA expression of hepatic insulin-like growth factor-1 (IGF-1) displayed a similar variation tendency as WG%. At the transcriptional level, the activity of Nrf2-Keap1 pathway was up-regulated by VC supplementations, with the VC104 treatment showing the highest value. In summary, the optimal dietary VC requirement for the highest value of WG% of juvenile hybrid grouper was 148 mg/kg dry matter, and appropriate dietary VC supplementation improved oxidation resistance and serum IgM concentrations of the fish. • VC affects growth and expression of hepatic IGF-1 of hybrid grouper. • At the transcriptional level, the activity of Nrf2-Keap1 pathway was up-regulated by VC supplementations. • Optimal VC requirement of hybrid grouper fed low fishmeal diets was 148 mg/kg. [ABSTRACT FROM AUTHOR]