Efficacy of Crystalline Lysine in Alternative Diets for Pond‐raised Hybrid Catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus

A study was conducted to examine the efficacy of crystalline lysine in alternative diets for pond‐raised hybrid catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus. Two 28% protein alternative diets supplemented with l ‐lysine HCl at the required level based on 62% (previously published value) or 100% lysine availability were compared with a traditional 28% protein control diet. Hybrid catfish fingerlings (mean initial weight = 43 g/fish) were stocked into 15 earthen ponds (0.04 ha) at a density of 14,826 fish/ha with five ponds per treatment. Fish were fed once daily to apparent satiation for a 173‐d growing season. There were no significant differences in total diet fed, net yield, weight gain, and survival among dietary treatments. There were also no significant differences in carcass yield, fillet yield, and fillet proximate composition and fillet lysine concentration among treatments. Fish fed the traditional control diet had slightly, but significantly, lower feed conversion ratio than fish fed alternative diets, which is likely related to higher dietary fiber levels in the alternative diets. Results from this study show that crystalline lysine can be considered 100% available when used to supplement lysine‐deficient diets for pond‐raised hybrid catfish.     

Pond‐raised Hybrid Catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus,Do Not Respond to Microbial Phytase Superdosing

Two experiments were conducted in consecutive years to evaluate the responses of hybrid catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus, to “superdosing” of 6‐phytase added to existing commercial catfish feeds. In each experiment, two diets with or without a phytase superdose (2500 and 5000 phytase units/kg, respectively) were compared. In Experiment 1, fingerlings (mean weight: 59 g/fish) were stocked in 17 0.4‐ha earthen ponds at 17,290 fish/ha and were fed once daily to apparent satiation for 198 d. In Experiment 2, fingerlings (mean weight: 47 g/fish) were stocked in 10 0.4‐ha ponds at 24,710 fish/ha and were fed for 128 d. In both experiments, there were no significant differences in total feed fed, gross yield, final fish weight, survival, or Blood packed cell volume between fish fed diets with or without phytase. The diets also had no significant effects on pond water column total phosphorus or chlorophyll a concentrations, but soluble reactive phosphorus concentrations were significantly higher in ponds receiving the phytase diet in Experiment 2. Phytase superdosing of nutritionally complete feeds does not appear to have additional benefits beyond the standard phytase dose on production characteristics or packed cell volume of pond‐raised hybrid catfish and had no beneficial effects on water quality.     

Water Quality and Plankton Communities in Hybrid Catfish (♀ Channel Catfish,Ictalurus punctatus × ♂ Blue Catfish,Ictalurus furcatus) Ponds After Partial Fish Harvest

Twelve 0.4‐ha ponds were stocked with 10,000 hybrid catfish fingerlings in March 2015. Six ponds were partially harvested in August to remove fish larger than ～0.57 kg. All remaining fish were removed in October and November. Partial harvest of faster‐growing fish removed ～26% of the fish initially stocked; feeding rate (and therefore external nutrient loading) was reduced by about the same amount. However, reduced nutrient loading after partial fish harvest had no meaningful effects on water quality, phytoplankton biomass, phytoplankton and zooplankton community structure, or supplemental aeration. Lack of ecosystem change was caused by persistent internal recycling of nutrients added to the system before partial harvest and continued high (albeit reduced) external nutrient loading after partial harvest. Decisions to employ partial fish harvest to manage single‐cropped hybrid catfish ponds should be based on economic considerations and risk reduction rather than the expectation that fish biomass reduction will improve water quality.     

Replacing Soybean Meal with Alternative Protein Sources in Diets for Pond‐raised Hybrid Catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus

The present study investigated the replacement of soybean meal with combinations of two or three alternative protein sources in diets for pond‐raised hybrid catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus. Alternative protein sources evaluated included cottonseed meal, distillers dried grains with solubles (DDGS), peanut meal, and porcine meat and bone meal (PMBM). Hybrid catfish fingerlings with a mean initial weight of 35 g/fish were stocked into 25 earthen ponds (0.04 ha) at a density of 14,826 fish/ha. Fish were fed once daily to apparent satiation for 166 d. No significant differences were observed for total diet fed, net yield, weight gain, survival, carcass yield, fillet yield, or fillet proximate composition among dietary treatments. Results show soybean meal may be completely replaced by combinations of cottonseed meal and one or two other alternative protein sources including DDGS, peanut meal, and PMBM in the diet without markedly affecting production and processing characteristics and fillet proximate composition of pond‐raised hybrid catfish. These alternative diets may be used during foodfish production when prices are favorable.     

Response of Pond‐raised Fingerling Hybrid Catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus,to Dietary Protein Concentrations and Sources

This study examined four experimental diets with different protein concentrations and sources for pond‐raised fingerling hybrid catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus. A 35% protein diet with fishmeal was used as the control diet. Test diets were 32 and 28% all‐plant‐protein diets and a 28% protein diet with porcine meat, bone, and blood meal. Small fingerlings with a mean initial weight of 2.9 g/fish were stocked into 20 earthen ponds (0.04 ha) at a density of 172,970 fish/ha. They were fed once daily to apparent satiation for 107 d. No significant differences were observed for total diet fed, gross yield, final weight, survival, or condition factor among dietary treatments. However, fish fed the 28 and 32% all‐plant‐protein diets had a significantly higher feed conversion ratio than fish fed the 35% protein diet with fishmeal. There were no significant differences in chlorophyll a and nitrite concentrations in the pond water, but ponds receiving the 35% protein diet had significantly higher ammonia than those receiving 28% protein diets. Economic analysis suggested potential cost savings by using low‐protein and all‐plant‐protein diets for hybrid catfish fingerling production.     

Emergence of Edwardsiella piscicida in Farmed Channel ♀, Ictalurus punctatus × Blue ♂, Ictalurus furcatus,Hybrid Catfish Cultured in Mississippi

There is a trend toward the increased incidence and prevalence of Edwardsiella piscicida septicemia in US catfish aquaculture, particularly in channel ♀, Ictalurus punctatus, × blue ♂, I. furcatus, hybrid catfish. From 2013 to 2017, a total of 3242 disease case submissions were made to the Aquatic Research and Diagnostic Laboratory (ARDL) at the Thad Cochran National Warmwater Aquaculture Center in Stoneville, MS. Of these, 1400 (43.2%) were hybrids. E. piscicida was suspected in 138 (4.3%) of cases, the majority of which (89.1%) were from hybrid catfish. A molecular survey of these isolates confirmed the majority (92.0%) to be E. piscicida. Furthermore, cases of E. piscicida from hybrids submitted to the ARDL and the Aquatic Diagnostic Laboratory of the Mississippi State University College of Veterinary Medicine in Starkville, MS, were documented for gross lesions and histological analysis. Grossly, E. piscicida presents with small dermal ulcerations, a raised fluid‐filled cranial midline lesion that is frequently ulcerated, hemorrhage in the gills, exophthalmia, and abdominal distension. Internally, lesions include splenomegaly, straw‐colored ascites, renomegaly, and occasionally hemorrhagic intestines. Histopathological examination is in agreement with gross observations, and infected fish repeatedly demonstrate a mononuclear meningoencephalitis, hemorrhagic branchitis, splenitis, ulcerative dermatitis, granulomatous interstitial nephritis, and hepatitis coupled with a hemorrhagic enteritis.     

Effects of Calcium and Magnesium Hardness of Incubation Waters on the Hatching Success of Hybrid Eggs of ♀ Channel Catfish,Ictalurus punctatus × ♂ Blue Catfish,Ictalurus furcatus

Hatching efficiency is generally lower in channel catfish, Ictalurus punctatus × blue catfish, Ictalurus furcatus, hybrid catfish hatcheries compared to hatcheries hatching pond‐spawned channel catfish eggs. Egg quality and hatching water hardness are known variables influencing the efficiency of hybrid catfish fry production in hatcheries. Previous research recommended that at least 60 mg/L hardness had to be contributed only by calcium to optimize the hatching success of hybrid catfish eggs. However, whether the effect of waterborne magnesium in hatching waters facilitates or impedes the hatching success of hybrid catfish is not known. Two trials were conducted to determine the effects of calcium alone or magnesium hardness, in combination with the hatching success of hybrid catfish eggs. Trial 1 showed that if the previously identified minimum hardness criterion of 60 mg/L was met by calcium in the hatching waters, added magnesium had no effect on the hatching success of hybrid catfish eggs. Trial 2 showed that the previously identified minimum hardness of 60 mg/L can be met by either calcium alone or magnesium replacing up to 50% of the total hardness in the hatchery waters for optimal hatching success of hybrid catfish eggs. Calcium provides a specific requirement during embryonic development of catfish. The results from this study can be used to design treatment processes for hatchery water supplies to improve efficiency of hybrid catfish fry production in hatcheries.     

Foodfish Production Strategies Using 13‐cm and 19‐cm Hybrid Catfish, ♀Ictalurus punctatus × ♂Ictalurus furcatus

Most hybrid catfish, ♀Ictalurus punctatus × ♂Ictalurus furcatus, producers stock 18‐ to 19‐cm fingerlings in single‐batch production. While stocking smaller fingerlings would be less expensive, the economic trade‐offs of using smaller fingerlings is unknown. Two sizes of hybrid catfish fingerlings (13‐cm and 19‐cm) were stocked in single‐size treatments at 9884 head/ha and a multi‐size treatment with each size stocked at 9884 head/ha (total 19,768 head/ha). In the 13‐cm and 19‐cm single‐size treatments, 87 and 98%, respectively, reached market size in one season. In the multi‐size treatment, 77% of all fish stocked reached market size, demonstrating that more than half of the 13‐cm fish reached market size in this treatment. Gross, net, and marketable yields were significantly greater in the multi‐size treatment when compared to the 13‐cm or 19‐cm single‐size treatments, but were not significantly different between the two single‐size treatments. The 19‐cm single‐size treatment resulted in greater break‐even prices and economic risk than the other two treatments. This study demonstrated that 13‐cm hybrid catfish fingerlings can be raised economically in both single‐ and multi‐size production systems in the southern USA.     

Comparative Susceptibility of Channel Catfish,Ictalurus punctatus; Blue Catfish,Ictalurus furcatus; and Channel (♀) × Blue (♂) Hybrid Catfish to Edwardsiella piscicida,Edwardsiella tarda,and Edwardsiella anguillarum

Members of the genus Edwardsiella are important pathogens of cultured and wild fish globally. Recent investigations into the phenotypic and genotypic variation of Edwardsiella tarda have led to the segregation of E. tarda into three distinct taxa: E. tarda, Edwardsiella piscicida, and Edwardsiella anguillarum. In catfish aquaculture in the southeastern USA, E. piscicida has been more commonly associated with disease than E. tarda or E. anguillarum, and recent research has demonstrated E. piscicida to be more pathogenic in channel catfish than E. tarda or E. anguillarum. Anecdotal reports from industry suggest an increased prevalence of E. piscicida associated with the culture of channel (♀) × blue (♂) hybrid catfish. This work investigated the comparative susceptibility of channel catfish, blue catfish, and their hybrid cross to molecularly confirmed isolates of E. tarda, E. piscicida, and E. anguillarum. There was significantly higher mortality in hybrid catfish compared to channel catfish following intracoelomic injection of E. piscicida. To our knowledge, E. piscicida is the first bacterial pathogen to demonstrate increased pathogenicity in hybrid catfish compared to channel catfish.     

Evaluation of Low‐protein Alternative Diets for Pond‐raised Hybrid Catfish,Ictalurus punctatus × Ictalurus furcatus

A study was conducted to evaluate low‐protein traditional or alternative diets for pond‐raised hybrid catfish, Ictalurus punctatus × Ictalurus furcatus. Three 24% protein diets containing decreasing levels of soybean meal (30, 20, and 15%) and increasing levels of cottonseed meal and corn germ meal were compared with a 28% protein control diet. Hybrid catfish fingerlings (mean initial weight = 71 g/fish) were stocked into 20 earthen ponds (0.04 ha) at a density of 14,826 fish/ha with five ponds per dietary treatment. Fish were fed once daily to apparent satiation for a 191‐d growing season. There were no significant differences in total diet fed, net yield, weight gain, feed conversion ratio (FCR), survival, or fillet proximate nutrient composition among dietary treatments (P ≥ 0.10). However, regression analysis showed for fish fed 24% protein diets there was a linear increase in FCR as soybean meal levels decreased (P = 0.06). Compared with fish fed the 28% protein control diet, fish fed 24% protein diets had lower carcass and fillet yield. Results demonstrate a 24% protein alternative diet containing 20% soybean meal may be substituted for 28% protein diets for hybrid catfish during food fish production.     

Performance of Alternative Diets Containing Solvent‐Extracted Distillers Dried Grains with Solubles Compared to Traditional Diets for Pond‐Raised Channel Catfish,Ictalurus punctatus,and Hybrid Catfish,Ictalurus punctatus × Ictalurus furcatus

Two concurrent pond studies were initiated in April 2011 to evaluate feeds containing solvent‐extracted distillers dried grains with solubles (SE‐DDGS) in both 28 and 32% protein catfish feeds. Channel catfish, Ictalurus punctatus, were stocked in multiple‐batch production ponds at the University of Arkansas at Pine Bluff (UAPB) and hybrid catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus, in single‐batch production ponds at the National Warmwater Aquaculture Center (NWAC), Mississippi State University, Stoneville, Mississippi, USA. The experimental design was a 2 × 2 factorial with two protein levels (32 and 28% protein), with and without 20% SE‐DDGS. Fish were fed once daily to apparent satiation, and managed similarly to a commercial farm for 186 d at UAPB and 172 d at NWAC. For channel catfish, two‐way ANOVA showed significant effects of both protein level (28% being better than 32%) and protein source (20% SE‐DDGS diets being better than traditional diets) on yield, survival, total diet fed, and feed conversion ratio (FCR), due primarily to effects on the understocked fingerlings. Partial budget analysis suggested that the 28% protein diet with the 20% SE‐DDGS was preferred economically for the prevailing feed costs. For hybrid catfish, two‐way ANOVA did not show significant differences in total diet fed, yield, survival, and processing yields, but use of SE‐DDGS resulted in a lower FCR. Results showed that both channel and hybrid catfish can utilize 20% SE‐DDGS in the diet without adversely affecting production or processing characteristics.     

The Influence of Ovarian Fluid pH of Stripped Unfertilized Channel Catfish,Ictalurus punctatus,Eggs on the Hatching Success of Channel Catfish ♀ x Blue Catfish,Ictalurus furcatus♂, Hybrid Catfish Eggs

This study was designed to determine the effect of ovarian fluid pH of stripped unfertilized channel catfish, Ictalurus punctatus, eggs on fertilization and hatch rate of channel catfish ♀ x blue catfish, Ictalurus furcatus♂, hybrid catfish eggs. A significant correlation was established between ovarian fluid pH of stripped channel catfish eggs and hybrid embryo hatch rate (R² = 0.75, P = 0.01), suggesting ovarian fluid pH of stripped catfish eggs prior to fertilization can be predictive of the hatching success of hybrid catfish embryos. These data were used to categorize pH of stripped eggs: pH <7.0 as “low pH eggs,” pH 7.0–7.4 as “medium pH eggs,” or pH >7.4 as “high pH eggs” quality eggs. The range in percent hatch rate for these pH categories was <15%, 15–30%, and >30%, respectively. Higher calcium concentrations during incubation do not appear to improve hatching success of “low pH eggs.” The predictive model presented herein describes a quick method for evaluating stripped catfish eggs for hybrid fry production in catfish hatcheries.     

Influence of forage fish and dietary lipid supplements on egg quality and fry production in channel catfish (Ictalurus punctatus) × blue catfish (Ictalurus furcatus) hybridization

Hybrid catfish (channel catfish Ictalurus punctatus × blue catfish Ictalurus furcatus) display characteristics that are favourable to aquaculture production. Low hatch percentages are a principal reason this hybrid is not used widely in the catfish industry. This study was conducted to determine whether additional food source rich in lipids may lead to a higher quality egg production. A 10‐week feed trial was conducted in ponds in Auburn, AL. A total of 219 female Kansas Select channel catfish were stocked into nine ponds, 0.04 ha in size. Three dietary treatments were randomly allocated to the ponds. Diet‐1 was a standard 60 g kg^?1 lipid floating catfish feed. Diet‐2 was the same feed supplemented with forage fish at ～28 kg ha^?1. The third diet was the aforementioned catfish feed topcoated with 20 g kg^?1 lipid [10 g kg^?1 menhaden fish oil, 5 g kg^?1 high docosahexaenoic acid (DHA) oil and 5 g kg^?1 high arachidonic acid oil]. Results indicate that brood fish fed the high lipid diet spawned larger egg masses and had larger eggs both in weight and in diameter, with increased complements of fatty acids such as DHA, eicosapentaenoic acid and total n‐3 fatty acids. The neutral and polar lipid fractions are also presented.     

The Costs and Effects of Alternative Winter Feeding Strategies for Channel Catfish Ictalurus punctatus in Multiple-Batch Production

Previous work suggested that feeding catfish, Ictalurus punctatus, more frequently over the winter might reduce weight loss. Twelve 0.10-ha ponds were stocked with 987 kg/ha market-sized fish and 2,960 kg/ha sub-marketable fish in November 2008 with treatments of: (1) unfed, (2) fed daily (fed 90 d), and (3) temperature-threshold feeding (fed 62 d). Total gross yield was significantly greater for the temperature-threshold feeding treatment than the unfed control, but survival and mean weight of fish at harvest were not. Net yield was negative for all treatments, due primarily to mortality of market-sized fish. Plasma glucose and insulin-like growth factor-1 did not differ significantly, but plasma osmolality was significantly lower in fasted than in fed fish. Mean feed consumption rates in vats provided evidence to support more frequent feeding over the winter. Overwintering costs were $0.11/kg with temperature-threshold feeding. Research on strategies to reduce winter mortality has potential to reduce overwintering costs.     

Characterization of triploid hybrid groupers from interspecies hybridization (Epinephelus coioides ♀ × Epinephelus lanceolatus ♂)

Interspecies hybridization is widely used in aquaculture as a beneficial strategy. Diploid and triploid hybrids have been detected from the interspecies hybridization of Epinephelus coioides ♀ × Epinephelus lanceolatus ♂. This is the first report of triploidization through hybridization in grouper. Confirmation has been obtained through flow cytometry, karyotyping and erythrocyte nuclei measurement. The chromosome numbers of E. coioides, E. lanceolatus, diploid hybrid grouper are 48 and triploid hybrid grouper are 72. Measurements of erythrocyte nuclei indicate that triploid fish have a larger nuclear surface than the diploid groupers, and the average ratio of triploid to diploid surface area is 1.59. During the first 1.5 years, triploid hybrid groupers grow faster than diploid hybrid groupers or either parent species. The average growth rate of triploid hybrids is 1.61 times greater than that of diploid hybrids at 6 months of age and 1.43 times greater at 18 months of age. The triploid hybrid groupers are inferior in gonadal development, with no primary‐growth‐stage oocytes appearing in the gonads at 18 months of age. Morphological studies indicate that triploid hybrid groupers have distinctive differences in snout length, eye diameter, body trunk shape, and tail shape development compared with diploid hybrid groupers. Triploid hybrid groupers have an advantage in growth ability, and artificial breeding of triploid groupers might be of great potential use in the grouper aquaculture industry.     

Dietary myo‐inositol requirements of juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂)

This study was conducted to determine the optimal dietary myo‐inositol (MI) requirement of juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂). Fish with an initial average body weight of 28.28 ± 0.41 g were randomly divided into six groups with three repetitions in each group and fed the experimental diets with different graded MI concentrations (70.2, 130.0, 270.2, 443.0, 1,096.6 and 1997.3 mg/kg) three times per day for 12 weeks. The results showed that with the increase in dietary MI level from 70.2 mg/kg to 443.0 mg/kg, weight gain (WG) was increased significantly (p < .05); however, when the MI level was further increased, there was no significant difference in weight gain rate (WG) over time (p > .05). Specific growth rate (SGR) showed a similar trend. The feed conversion ratio (FCR), survival rate (SR), hepatosomatic index (HSI), viscerasomatic index (VSI) and condition factor (CF) were not affected by dietary MI supplementation (p > .05). Dietary MI did not affect whole‐body and muscle crude protein and ash content (p > .05), while dietary MI significantly affected the muscle and whole‐body crude lipid and moisture content (p < .05). Moreover, serum triglyceride (TG), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were decreased by increasing dietary MI levels. Total antioxidant capacity (T‐AOC), malonic dialdehyde (MDA) and superoxide dismutase (SOD) activities in the liver were also affected by the dietary MI (p < .05). In summary, this study indicated that the optimal dietary MI requirement for juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂) was 335.84 mg/kg and 590.65 mg/kg based on the maximum WG and liver MI accumulation indices respectively.     

Responses of hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenckii ♂) to oral administration of phosphorus

To better understand the phosphorus (P) utilization in hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenckii ♂), a combined technique with oesophageal intubation, dorsal aorta cannulation and urinary catheterization was used in the present study. Groups of five hybrid sturgeon were orally administrated with graded doses of P (0, 50, 100 and 200 mg P/kg BW) in the form of monocalcium phosphate. The high dose of P (200 mg P/kg BW) caused a significant increase in plasma P concentration. Plasma calcium (Ca) concentrations were significantly increased in the fish treated with 0 and 100 mg P/kg BW. Both P and urea excretion showed increased changing pattern with increasing P intubation level and exhibited significant higher value in 200 mg P/kg BW group. A significant increase in Ca excretion was found in the group without P supplementation (0 mg P/kg BW). ALP and SOD activity in plasma was inhibited in fish at P‐deficient groups (0 and 50 mg P/kg BW). The data in the current study showed that hybrid sturgeon at 100 mg P/kg BW treatment exhibited better P and Ca utilizations with lower waste excretion, as well as favourable status of anti‐oxidation.     

Ploidy polymorphism and morphological variation among reciprocal hybrids of Pseudosciaena crocea (♀) × Miichthys miiuy (♂)

Diploid and triploid hybrids of large yellow croaker (Pseudosciaena crocea, Pc) and Miiuy croaker (Miichthys miiuy, Mm) were produced by interspecific hybridization. The result showed that Pc (♀) and Mm (♂) can be successfully crossed and hybrid offspring of normal hatch rate were produced. In contrast, a lower fertilize and hatch rate were produced for hybrids of Pc (♂) and Mm (♀), and the offspring died day 19 after hatching. To confirm the ploidy of hybrid offspring, we examined the karyotypes of offspring. The result of DNA relative content investigation and genetic relationship analysis demonstrated that the major hybrid offspring were hybrid diploid. In addition, a small number of hybrid were triploid and gynogenesis diploid. Morphological analysis based on the measurable traits data indicated the success of the interspecific crosses. The natural gynogenetic diploid large yellow croaker (PP), diploid hybrids (PM) and triploid hybrids (PPM) were closer to maternal parent (Pc) than paternal parent (Mm). This study provided genetic evidences of the chromosome, DNA content and morphological levels to support the successful establishment of the polyploidy hybrids of Pc (♀) and Mm (♂).     

The optimum dietary isoleucine requirement of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)

A six‐week growth trial was conducted to evaluate the optimum dietary isoleucine requirement of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). Seven isoenergetic (3,400 kcal/kg of dry matter), isoproteic (496 g/kg of dry matter) and isolipidic (70 g/kg of dry matter) diets were formulated to contain graded Ile levels (7.3, 11.3, 15.7, 19.6, 23.5, 26.8 and 30.8 g/kg, dry‐matter basis). Each experimental diet was fed to triplicate groups of 12 hybrid grouper juveniles (average initial body weight: 6.00 ± 0.01 g/fish). Experimental fish were randomly distributed into 21 glass tanks (L 60 × W 45 × H 50 cm) connected to mechanical and biological water filters as a recycling system. Fish were fed twice daily (08:00 and 16:00) to apparent satiation. After the sampling of the growth trial, the remaining fish in each group were fed their corresponding diets for 2 d and then exposed to 4 mg Cu²⁺ · L^?1 water for 24 hr. Results showed that growth performance and feed utilization were significantly affected by different dietary Ile levels (p < .05). Weight gain percentage (WG%), protein productive value (PPV), protein efficiency ratio (PER) and feed efficiency (FE) were increased as dietary Ile level increased, reaching a peak value at 19.6 g/kg dietary Ile, and thereafter, these four parameters declined as dietary Ile level continued to increase. Daily feed intake (DFI) showed an opposite tendency of variations as FE. The quadratic regression analysis of WG%, PPV, PER and FE against dietary Ile levels indicated that the optimum dietary Ile requirement for hybrid grouper was estimated to be 19.8, 20.8, 19.4 and 19.1 g/kg dry matter, respectively. Among all experimental treatments, fish fed 19.6 g/kg dietary Ile had the highest expression of growth and protein synthesis‐related genes, including growth hormone (GH) in pituitary, insulin‐like growth factor 1 (IGF‐1), growth hormone receptor 1 (GHR1), target of rapamycin (TOR) and S6‐kinase 1 (S6K1) in liver. Gut micromorphology was significantly influenced by dietary Ile levels. After the exposure to 4 mg Cu²⁺ · L^?1 water for 24 hr, fish fed 19.6 g/kg dietary Ile had the highest survival and the best immunologic manifestation among all experimental treatments. Generally, the optimum dietary Ile requirement for maximum growth of hybrid grouper was estimated to be 19.8 g/kg dry matter, corresponding to 39.9 g/kg dietary protein.