Dietary potassium requirement of fingerling Indian major carp,Labeo rohita (Hamilton), based on growth parameters,gill Na+‐K+ ATPase activity,tissue mineralization and antioxidant activities

To quantify the dietary potassium requirement of fingerling Labeo rohita (6.2 ± 0.12 cm; 1.98 ± 0.06 g), seven purified experimental diets (350 g/kg crude protein and 16.72 kJ/g gross energy) with graded levels of potassium (0.32, 1.35, 2.41, 3.46, 6.48, 9.47 and 12.39 g/kg diet) were fed to triplicate groups of fishes at 08:00, 12:00 and 16:00 hr to apparent satiation for 8 weeks. Live weight gain (LWG; 671.46%), specific growth rate (3.65%/day), protein efficiency ratio (2.16), protein gain (PG; 2.41 g/fish) and feed conversion ratio (1.32) were found to be best in fish fed diet containing 3.46 g/kg potassium. Gill Na⁺‐K⁺ ATPase activity was also highest in fish fed diet with 3.46 g/kg potassium. Potassium content of whole‐body, vertebrae and scales increased significantly with the increase in dietary potassium level up to 6.48 g/kg. Significant changes were also noted in serum malondialdehyde content, superoxide dismutase, catalase, glutathione peroxidase and alkaline phosphatase activity. Based on the maximum live weight and protein gain observed in the present study, the inclusion of 3.55 g/kg potassium is recommended for developing potassium‐balanced commercial feeds for intensive culture of fingerling L. rohita.     

Dietary folic acid requirement of fingerling Catla,Catla catla (Hamilton)

The dietary folic acid requirement of fingerling Catla catla (3.4 ± 0.17 g; 7.6 ± 0.41 cm) was evaluated by feeding casein–gelatin‐based isonitrogenous (350 g/kg crude protein) and isocaloric (16.72 kJ/g GE) diets containing different concentrations of folic acid (0, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0 mg/kg) to triplicate groups to apparent satiation at 08:00, 12:30 and 17:30 hr for 16 weeks. Absolute weight gain (AWG; 40.07 g/fish), specific growth rate (SGR; 2.25%), feed conversion ratio (FCR; 1.53), protein retention efficiency (PRE; 31.42%) and protein gain (PG; 6.74) improved significantly (p < .05) with increasing folic acid levels up to 0.4 mg/kg diet and then reached a plateau. However, maximum liver folic acid concentration increased up to 0.6 mg/kg diet. Dietary folic acid levels also significantly affected (p < .05) body composition of fish. No significant change (p > .05) in haematological parameters except in fish fed folic acid‐free diet was noted. Antioxidant and immune parameters increased with increasing concentration of dietary folic acid up to 0.4 mg/kg diet. Broken‐line regression analysis of AWG, FCR, PRE, PG, HCT and liver folic acid concentrations of fingerling C. catla against dietary folic acid levels indicated optimum growth, FCR, PRE, PG, HCT and liver folic acid saturation ranging between 0.22 and 0.56 mg/kg diet, respectively.     

Optimum level of dietary monocalcium phosphate determined based on optimal growth and vertebrae phosphate content of juvenile Ussuri catfish,Pseudobagrus ussuriensis

Two experiments were conducted to determine the optimum level of dietary available phosphorus from monocalcium phosphate for juvenile Ussuri catfish Pseudobagrus ussuriensis. Experiment 1 was conducted to estimate phosphorus digestibility from monocalcium phosphate for juvenile Ussuri catfish. The apparent digestibility coefficient of phosphorus from monocalcium phosphate was 86.3%. In the experiment 2, triplicate groups of juvenile Ussuri catfish were fed diets containing graded levels of monocalcium phosphate (MCP: 0 g/kg, 8.2 g/kg, 16.4 g/kg, 24.6 g/kg, 32.8 g/kg and 41.0 g/kg) for 8 weeks. Fish fed the diet containing 16.4 g/kg MCP with available phosphorus of 4.8 g/kg showed the best weight gain (171.5%), feed conversion ratio (1.08) and protein efficiency ratio (2.06). No significant difference was observed in fish survival among the treatments. The best result in terms of phosphorus retention efficiency (46.10%) was observed in fish fed the diet containing 8.2 g/kg MCP with available phosphorus of 3.0 g/kg, which was not different (p > .05) from those fed the diet containing up to 24.6 g/kg MCP, and the highest vertebrae phosphorus content (58.2 g/kg) was observed in fish fed the diet containing 24.6 g/kg MCP with available P of 6.6 g/kg. The whole‐body lipid and protein, as well as phosphorus contents, were significantly affected by dietary available phosphorus (p < .05). Viscerosomatic index (VSI) and condition factor (CF) were inversely correlated with dietary phosphorus levels (p < .05). Quadratic regression analysis based on specific growth rate (SGR) against dietary available phosphorus levels indicated that the optimum available phosphorus requirement for the maximal growth of juvenile Ussuri catfish was 5.9 g/kg, and broken‐line analysis based on vertebrae phosphorus content against dietary available phosphorus levels indicated that a dietary level of 6.0 g/kg available phosphorus will provide optimum vertebrae phosphorus content.     

Determination of dietary phosphorus requirement of stinging catfish Heteropneustes fossilis based on feed conversion,growth, vertebrae phosphorus,whole body phosphorus,haematology and antioxidant status

A 12‐week feeding trial was conducted to determine the dietary phosphorus requirement of Heteropneustes fossilis fingerlings (7.7 ± 0.04 g). Fish were fed casein–gelatine‐based purified diets in triplicate groups near satiation with seven different levels of dietary phosphorus (3.2, 5.2, 7.2, 9.2, 11.2, 13.2 and 15.2 g/kg dry diet). All diets were formulated to be isoproteic (400 g/kg) and isoenergetic (17.89 kJ/g). Highest absolute weight gain (68.38 g/fish), best feed conversion ratio (1.48), protein retention efficiency (30.74%), protein gain (12.44 g/fish), haemoglobin (11.19 g/dL), RBCs (3.12 x10⁶/mm³), haematocrit (33.44%) and serum phosphate (2.82 mg/L) were found at 9.2 g/kg phosphorus. Hepatic superoxide dismutase and catalase activity were also significantly influenced by the dietary phosphorus levels. Whole body and vertebrae phosphorus concentrations increased significantly as the amount of dietary phosphorus increased from 3.2 to 11.2 g/kg dry diet and then plateaued. More accurate information on dietary phosphorus requirement was obtained by subjecting the AWG, FCR, vertebrae phosphorus and whole body phosphorus concentrations data against various levels of dietary phosphorus to broken‐line analysis, which yielded the requirement in the range of 9.0–11.0 g/kg for optimum growth and mineralization of H. fossilis.     

Dietary thiamin requirement of fingerling Channa punctatus (Bloch) based on growth,protein gain,liver thiamin storage,RNA/DNA ratio and biochemical composition

Dietary thiamin requirement of fingerling Channa punctatus was quantified by feeding casein/gelatin‐based diets (450 g/kg CP; 18.39 kJ/g GE) with seven graded levels of thiamin (0, 0.5, 1, 1.5, 2, 2.5 and 5 mg/kg diet) to triplicate groups of fish (6.9 ± 0.93 cm; 4.91 ± 0.62 g) for 16 weeks. Fish fed diet with 2.5 mg/kg thiamin reflected highest absolute weight gain (AWG), protein gain (PG), RNA/DNA ratio and lowest feed conversion ratio. Similarly, highest liver thiamin concentration was also recorded in fish fed 2.5 mg/kg thiamin diet. Hepatic thiobarbituric acid reactive substance (TBARS) concentration responded negatively with increasing concentrations of dietary thiamin up to 2.5 mg/kg, whereas superoxide dismutase and catalase activities were found to improve with the increasing levels of dietary thiamin from 0 to 2.5 mg/kg. Transketolase activity also improved as the thiamin concentrations increased up to 2.5 mg/kg. Broken‐line regression analysis of AWG, PG, RNA/DNA ratio, liver thiamin concentrations, transketolase and TBARS activities exhibited the thiamin requirement in the range of 2.34–2.59 mg/kg diet. Data generated during this study would be useful in formulating thiamin‐balanced feeds for the intensive culture of this fish.     

Dietary riboflavin requirement of fingerling Channa punctatus (Bloch) based on growth,conversion efficiencies,protein retention,liver riboflavin storage,RNA/DNA ratio and carcass composition

A 16‐week experiment was conducted to determine the dietary riboflavin requirement of the fingerling Channa punctatus (6.7 ± 0.85 cm; 4.75 ± 0.72 g) by a feeding casein–gelatin‐based (450 g/kg crude protein; 18.39 kJ/g gross energy) purified diet containing graded levels of riboflavin (0, 2, 4, 6, 8, 10 and 12 mg/kg diet) to triplicate groups of fish near to satiation at 09:30 and 16:30 hr. Absolute weight gain (AWG), protein efficiency ratio (PER), specific growth rate (SGR, % per day), protein retention efficiency (PRE%) and RNA/DNA ratio were positively affected by increasing concentrations of dietary riboflavin to 6 mg riboflavin per kg diet. Feed conversion ratio (FCR) decreased up to 6 mg riboflavin per kg diet but did not decrease further with higher riboflavin supplementation. Hepatic thiobarbituric acid‐reactive substance (TBARS) concentration also supported the pattern of FCR, whereas superoxide dismutase and catalase activities increased with increasing concentrations of dietary riboflavin from 0 to 6 mg/kg. Liver riboflavin concentrations increased with increasing levels of riboflavin up to 8 mg/kg diet. Broken‐line regression analysis of AWG, PRE and liver riboflavin concentrations of fingerling C. punctatus with dietary riboflavin level indicated optimum growth and liver riboflavin saturation at 5.7, 6.1 and 7.7 mg riboflavin per kg diet, respectively.     

The effects of dietary inulin and Jerusalem artichoke (Helianthus tuberosus) tuber on the growth performance,haematological, blood chemical and immune parameters of Nile tilapia (Oreochromis niloticus) fingerlings

This study investigated the effects of dietary inulin or Jerusalem artichoke (JA) on the growth performance, haematological, blood chemical and immune parameters of Nile tilapia fingerlings. Five treatment diets were designed to incorporate inulin at 0 (basal diet), 2.5 and 5.0 g/kg and JA at 5.0 and 10.0 g/kg. Two basal diets including fish meal and formulated experimental feed were used for fry and fingerling growing periods, respectively. During the fry growing period, larvae were fed treatment diets for 4 weeks. There were no significant differences in growth performance or survival rate. Fingerlings were then nursed with the formulated experimental diets from weeks 5 to 12. Fingerlings fed on inulin at 5.0 g/kg or JA at either level had better growth performance and survival rate than that fed on the basal diets. There were no significant differences in body composition. Dietary prebiotic inulin and JA increased red blood cell number (p < .05). Among the five blood chemistry parameters examined, both inulin at 5.0 g kg and JA (5.0 and 10.0 g/kg) increased blood protein (p < .05). Dietary inulin at 5.0 g/kg and JA at 5.0 and 10.0 g/kg increased total immunoglobulin and lysozyme activity (p < .05). Both inulin and JA inclusion diets increased alternative complement activity (p < .05). Taken together, dietary inulin at 5 g/kg and JA at 5.0 and 10.0 g/kg had beneficial effects on the growth performance, survival rate and immune of Nile tilapia fingerlings.     

A comparison of the effect of organic acids and dicalcium phosphate supplementation on phosphorus bioavailability,growth performance and digestive enzyme activities of Labeo rohita fingerlings

The effect of organic acids (OA) and dicalcium phosphate (P_i) supplementation in the feed of Labeo rohita fingerlings was studied by formulating seven practical diets, designed as D1 with no feed additives {dicalcium phosphorus (P_i) and organic acid (OA) blend} while, D2, D3, D4 and D5 were supplemented with P_i at the graded levels of 5, 10, 15 and 20 g/kg, respectively, whereas D6 and D7 contained blend of OA at 15 and 30 g/kg, respectively. The diet supplemented with OA blend exhibited increase (p < .05) in growth of L. rohita fingerlings compared with the diet supplemented with P_i. The OA supplementation at both levels showed significant improvement in nutrients digestibility and minerals absorption in fingerlings. Moreover, in case of muscle proximate composition, crude fat (CF) and crude protein (CP) were increased (p < .05) with the inclusion of OA blend in the diet while crude ash (CA) was improved by P_i supplementation. Additionally, digestive enzyme activities were not affected (p ? .05) by OA blend supplementation while increased activities were observed in the fingerlings fed with P_i diet. Consequently, supplementation of OA blend in the diet improved the growth performance and nutrient status while P_i enhanced digestive enzyme activities of L. rohita fingerlings.     

Dietary available phosphorus requirement of crucian carp,Carassius auratus

This study was carried out to evaluate the dietary available phosphorus (AP) requirement for crucian carp (Carassius auratus). Triplicate groups were fed diets containing five graded levels of AP in 15 recirculating tanks. After a 9‐week feeding experiment, weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and whole‐body and vertebrae P contents were significantly increased as dietary AP increased from 1.1 to 7.6 g/kg (p < .05) and then levelled off. N and P retention was also significantly increased as dietary AP increased to 5.5 g/kg (p < .05). Condition factor, viscerosomatic index, hepatosomatic index, whole‐body moisture, muscle P content and plasma total cholesterol were not affected by dietary P levels (p > .05). Protein and ash contents of the whole body increased linearly as the dietary P level increased, but the lipid content, plasma alkaline phosphatase activities and triacylglycerol contents showed an inverse relationship. Based on WG, FE, whole‐body P content and vertebrae P content, the optimum requirement of dietary AP for crucian carp was estimated to be 8.3, 8.3, 8.0 and 7.8 g/Kg, respectively.     

Dietary available phosphorus requirement for juvenile gibel carp (Carassius auratus gibelio var. CASIII)

A 57‐day growth experiment was conducted with juvenile gibel carp (13.48 ± 0.10 g) in a flow‐through system to study the effect of dietary phosphorus on growth performance, body composition, nutrition utilization, phosphorus loading and enzymes activities. Seven semipurifed diets were formulated to contain 0.07 (the basal), 2.27, 5.32, 8.10, 12.06, 15.24 and 19.48 g available phosphorus/kg diet. The results showed that specific growth rate, body length and feed efficiency significantly increased in the fish fed diets containing 0.07 to 15.24 g available P/kg diet (p < .05). Ash and P content increased in fish fed diets containing 0.07–12.06 P g/kg (p < .05) and then levelled off, while moisture, crude protein and lipid had no significant difference (p > .05). The protein retention efficiency increased in the fish fed with diets 0.07–5.32 g/kg P (p < .05) and then reached a plateau. The P content in faeces was higher in fish fed diets containing 15.24 and 19.48 g available P/kg. Total P concentration in tank water increased in fish fed 0.07–12.06 g available P per kg diet (p < .05). The plasma P was higher in the fish fed with 15.24 g available P/kg diet (p < .05), triglycerides was lower in the fish fed diet containing 15.24 and 19.48 g available P/kg (p < .05), no significant differences were observed in plasma Ca, plasma glucose and calcitonin (p > .05). Based on SGR, whole body P content and FE, dietary available P requirement for juvenile gibel carp were 13.37, 13.97 g/kg and 15.06 respectively.     

Dietary biotin requirement of fingerling Channa punctatus (Bloch)

ABSTRACT

Dietary biotin requirement of fingerling Channa punctatus (4.52 ± 0.46 g) was estimated by conducting a 16-week growth trial. Fish were fed casein gelatin-based purified diets (450 g/kg crude protein, 18.39 kJ/g gross energy) with seven levels of dietary biotin (0, 0.04, 0.09, 0.47, 1.02, 1.43, and 1.96 mg/kg diet) to triplicate groups near to satiation. Significantly higher absolute weight gain (P = 0.0018), specific growth rate (P = 0.0027), protein gain (P = 0.0016), protein deposition (P = 0.0038), and lower feed conversion ratio (P = 0.0003) were shown in fish fed diet containing 0.47 mg/kg biotin, whereas liver biotin concentration showed a significant improvement (P = 0.0021) with increasing levels of dietary biotin up to 1.02 mg/kg. Broken-line analysis of absolute weight gain, protein gain, and liver biotin concentrations indicated that fingerling C. punctatus require biotin at 0.46, 0.44, and 0.97 mg/kg diet. Based on protein gain, optimum pyridoxine requirement for fingerling C. punctatus is recommended at 0.44 mg/kg diet.     

Effect of dietary niacin on growth and body composition of two Indian major carps rohu, <Emphasis Type="Italic">Labeo rohita</Emphasis>, and mrigal, <Emphasis Type="Italic">Cirrhinus mrigala</Emphasis> (Hamilton), fingerlings based on dose–response study

An 15 week two set of feeding experiments were conducted to determine the dietary niacin requirement of Indian major carp fingerlings Labeo rohita and Cirrhinus mrigala, using casein gelatin–based diet. In both experiments, six isonitrogenous (40%) and isoenergetic (15.35 kJ g⁻¹) test diet, with graded levels of niacin (0–50 mg kg⁻¹ dry diet) in gradation of 10 mg kg⁻¹ dry diet, were formulated. In first experiment, fingerling of L. rohita (4.20 ± 1.22 cm; 0.632 ± 0.67 gm) were randomly stocked, in triplicate groups, in 55-L indoor polyvinyl flow-through system (1.5 L min⁻¹) and fed experimental diet at 0800 and 1600 h. Maximum live weight gain (1214%), feed conversion ratio (1.55) and protein efficiency ratio (1.60) were recorded at 30 mg dietary niacin diet. In second experiment, C. mrigala (4.50 ± 1.25 cm, 0.665 ± 0.88) were stocked in same setup. At the end of experiments, maximum live weight gain (1248%), FCR (1.47) and PER (1.70) occurred at 30 mg dietary niacin diet. However, the weight gain, FCR and PER data were analyzed by polynomial regression analysis indicating the requirement of niacin for L. rohita at 36.69, 33.06 and 32.0 mg kg⁻¹, respectively, and for C. mrigala at 35.19, 28.69 and 27.70 mg kg⁻¹ of dry diet, respectively. Whole body composition also showed significant (P < 0.05) differences among each other. On the basis of regression analysis of growth data, it is recommended that the diet for fingerlings should contain niacin at 33 and 30 mg kg⁻¹ dry diet for L. rohita and C. mrigala, respectively.     

Effects of glutamate in low‐phosphorus diets on growth performance,antioxidant enzyme activity,immune‐related gene expression and resistance to Aeromonas hydrophila of juvenile mirror carp (Cyprinus carpio)

This study investigated the effects of glutamate (Glu) in low‐phosphorus diets on growth performance, haematological indices, antioxidant enzyme activity, immune‐related gene expression and resistance to Aeromonas hydrophila in juvenile mirror carp (Cyprinus carpio) (5.07 ± 0.02 g). Fish were fed either graded levels of Glu (0 g/kg, 5 g/kg,  10 g/kg and 20 g/kg, named G0, G0.5, G1 and G2, respectively) in a low‐phosphorus diet (15 g/kg NaH₂PO_4, 0.49), or a normal phosphorus diet ( 20 g/kg NaH₂PO_4, 0.61) without added Glu (C), for 8 weeks. At the end of the feeding trial, the fish were challenged with A. hydrophila. Compared with G0 group, 10 g/kg and 20 g/kg Glu supplementation of the low‐phosphorus diet significantly improved the final weight, WGR, SGR and PER, and decreased FCR (p < .05). Glu supplementation of the low‐phosphorus diet significantly enhanced the T‐AOC, SOD activity and GSH content in intestine (p < .05). Glu supplementation significantly reduced MDA content in foregut and midgut and increased CAT activity in midgut and hindgut (p < .05). Regarding immune‐related gene expression, Glu supplementation significantly diminished the up‐regulation of intestinal TNF‐α, IL‐1β and IL‐8 mRNA levels induced by phosphorus deficiency (p < .05). The survival rate of the G1 group was significantly higher than that of the G0 group (p < .05). In conclusion, 10 g/kg Glu supplementation in low‐phosphorus diets can improve the growth performance, enhance the activity of intestinal antioxidant enzymes and strengthen the immune function of juvenile mirror carp.     

Dietary Arginine Requirement of Fingerling Indian Major Carp,Catla catla (Hamilton)

Dietary arginine requirement of fingerling Catla catla (3.55 ± 0.05 cm; 0.61 ± 0.02 g) was determined by feeding casein–gelatin‐based isonitrogenous (33% crude protein) and isocaloric (3.40 kcal/g digestible energy) amino acid test diets containing six graded levels of l ‐arginine (1, 1.25, 1.5, 1.75, 2, and 2.25% dry diet) for 12 wk. Maximum absolute weight gain (6.93 g/fish), protein efficiency ratio (2.13), protein deposition (0.36), arginine retention efficiency (78%), and best feed conversion ratio (1.42) were recorded in fish fed 1.75% arginine of the dry diet. Maximum carcass protein (15.57%) and RNA/DNA ratio (4.79) were also recorded for the group fed 1.75% arginine of the dry diet. Quadratic regression analysis at 95% maximum or minimum response of above growth parameters yielded optimum arginine requirement of fingerling C. catla at 1.67% of the dry diet. On the basis of the above analysis of the growth parameters, it is recommended that the inclusion of dietary arginine at 1.67% of the dry diet is optimum for formulating arginine‐balanced, cost‐effective quality feeds for the mass culture of fingerling C. catla .     

Utilization of electron beam irradiated Jatropha kernel meal in the diet of Labeo rohita (Hamilton, 1822) fingerlings

Defatted Jatropha kernel meal (DJKM) was irradiated through electron beam radiation at 25 kGy (IJKM). After irradiation, PEs and phytate were decreased by 36.67% and 55.27%, respectively, with slight reduction in total hydrolysed amino acids in IJKM. A 45‐day feeding trial was conducted to evaluate the utilization of irradiated Jatropha kernel meal (IJKM) in the diet of rohu (Labeo rohita) fingerlings. Five isonitrogenous (300 g/kg CP) and isoenergetic (15 MJ/kg GE) diets such as T0 (control, without IJKM), T5 (50 g/kg IJKM), T10 (100 g/kg IJKM), T15 (150 g/kg IJKM) and T20 (200 g/kg IJKM) were prepared and fed to fish of respective treatments. Fish fed diets containing T15 and T20 groups exhibited significantly lower (p < .05) weight gain, FCE, PER, ANPU, HSI, ISI, survival rate, nutrient and energy digestibility, than the other groups. Fish of higher IJKM fed groups (T15 and T20) also showed lower muscle moisture, protein, ash and higher muscle lipid content. The liver catalase and SOD activities significantly decreased in the higher IJKM fed groups. It is concluded that IJKM (irradiated by 25 kGy electron beam) can be incorporated up to 100 g/kg in carp feed with the replacement of 33% soybean meal and 28% ground nut oil cake without compromising growth performances of Labeo rohita.     

Dietary pantothenic acid requirement of fingerling Channa punctatus (Bloch) based on growth,feed conversion,liver pantothenic acid concentration and carcass composition

A 16‐week feeding trial was conducted to determine the dietary pantothenic acid requirement of fingerling Channa punctatus. Six casein–gelatin‐based diets (450 g/kg CP; 18.39 kJ/g GE) with graded levels of pantothenic acid (0, 10, 20, 30, 40 and 50 mg/kg diet) were fed to triplicate groups of fish (6.2 ± 0.71 cm; 4.26 ± 0.37 g) near to apparent satiation. The growth evaluation in terms of absolute weight gain (AWG), feed conversion ratio (FCR) and protein retention efficiency (PRE) indicated the best performance (p < .05) in fish fed diet containing 30 mg/kg pantothenic acid. Highest haemoglobin, haematocrit and RBCs counts were also obtained in fish fed diet with 30 mg/kg pantothenic acid. Mean cell haemoglobin and mean cell volume were found to be lowest in fish fed pantothenic acid‐free diet indicating the anaemia in this group of fish. Superoxidase dismutase and catalase activities of liver tissue were found to improve (p < .05) with the increasing levels of dietary pantothenic acid from 0 to 30 mg/kg. However, liver pantothenic acid concentration responded positively with the increasing levels of pantothenic acid up to 40 mg/kg diet and then stagnation in liver pantothenic acid concentration with the further inclusion of pantothenic acid was recorded. Second‐degree polynomial regression analysis of AWG, FCR and PRE exhibited the pantothenic acid requirement at 36.4, 32.8 and 34.7 mg/kg diet, respectively. Data generated during this study would be useful in formulating pantothenic acid‐balanced commercial feeds for the intensive culture of this fish.     

Effects of dietary inulin and Jerusalem artichoke (Helianthus tuberosus) on intestinal microbiota community and morphology of Nile tilapia (Oreochromis niloticus) fingerlings

This study investigated the effects of dietary inulin and Jerusalem artichoke (JA) on intestinal microbiota and morphometry of Nile tilapia fingerlings. Five treatment diets were designed to supplement inulin at 0 (basal diet), 2.5 and 5.0 g/kg, and JA at 5.0 and 10.0 g/kg. Nile tilapia larvae were fed experimental diets from the first feeding through the fingerling stage (84 days). The cultivation‐dependent technique showed that dietary inulin at 5.0 g/kg and JA (at both levels) increased lactic acid bacteria and Bifidobacterium spp., but decreased Vibrio spp. (p < .05). PCR‐DGGE targeting 16S ribosomal RNA gene revealed that dietary inulin and JA generated different profiles of microbial community compared with fish fed a basal diet. Compared with fish fed the basal diet, a greater intestinal villi height was observed in fish fed 5.0 g/kg inulin and JA at both levels (p < .05). A larger relative goblet cell number were observed in the anterior intestine of fish fed 5.0 g/kg inulin or JA (p < .05). Overall, dietary inulin (5.0 g/kg) and JA (5 and 10.0 g/kg) since the first feeding had effects on modulating the intestinal microbiota and morphology of Nile tilapia fingerlings.     

Dietary copper requirements of juvenile grouper, Epinephelus malabaricus

To quantify dietary copper (Cu) requirements in grouper, Epinephelus malabaricus, copper sulfate was added to the basal diet at 0, 1, 2, 4, 6, 8, 10 and 20 mg Cu/kg diet providing the actual dietary value of 0.11, 1.66, 2.41, 4.37, 6.56, 8.97, 11.03 and 20.05 mg Cu/kg diet, respectively. Each diet was fed to triplicate groups of grouper (initial body weight 13.35 ± 0.01 g) in a recirculated rearing system for 8 weeks. The Cu concentration in rearing water was monitored during the feeding period and ranged from 1.0-1.5 μg/l. Weight gain (WG) and feed efficiency (FE) were higher (P < 0.05) in fish fed diets with 4.37 and 6.56 mg Cu/kg diet than those in fish fed diets with ≥ 11.03 and ≤ 1.66 mg Cu/kg diet. Hepatic thiobarbituric acid reactive substances (TBARS) value was lowest in fish fed diets with 4.37 and 6.56 mg Cu/kg diet, intermediate in fish fed diets with 11.03 and ≤ 1.66 mg Cu/kg diet, and highest in fish fed the diet with 20.05 mg Cu/kg diet. Differences between each of the three groups were statistically significant (P < 0.05). Both hepatic copper-zinc superoxide dismutase (Cu-Zn SOD) and plasma ceruloplasmin activities in fish fell into four groups. Cu-Zn SOD value was highest in fish fed the diet with 6.56 mg Cu/kg diet, second highest in fish fed the diet with 8.97 mg Cu/kg diet, intermediate in fish fed the diet with 20.05 mg Cu/kg diet, and lowest in fish fed the diet with 0.11 mg Cu/kg diet; plasma ceruloplasmin was highest in fish fed the diet with 6.56 mg Cu/kg diet, second highest in fish fed diets with 2.41, 4.37 and 8.97 mg Cu/kg diet, intermediate in fish fed diets with ≥ 11.03 mg Cu/kg diet, and lowest in fish fed diets with ≤ 1.66 mg Cu/kg diet. All values in each of the four groups were significantly different from the values of three other groups (P < 0.05). Analysis by broken-line regression of WG, hepatic SOD activity and TBARS value and linear regression of whole-body Cu retention of the fish indicate that the adequate dietary Cu concentration in growing grouper is about 4-6 mg Cu/kg diet.     

Dietary Houttuynia cordata leaf extract and meal enhances the immunity and expression of immune genes in Labeo rohita (Hamilton, 1822)

A feeding trial was conducted to explore the effect of dietary Houttuynia cordata leaf extract (HCLE) and leaf meal (HCLM) on immunological responses and expression of interferon‐gamma (IFN‐γ) and tumour necrosis factor‐alpha (TNF‐α) gene in Labeo rohita fingerlings. Six isonitrogenous (350 g/kg CP) and isocaloric (17 MJ/kg DE) purified experimental diets were formulated with Houttuynia cordata leaf extract and leaf meal comprising control, C (0 g/kg HCLE and HCLM), E2.5 (2.5 g/kg HCLE), E5 (5 g/kg HCLE), E10 (10 g/kg HCLE), M10 (10 g/kg HCLM) and M20 (20 g/kg HCLM). Labeo rohita fingerlings (3.37 ± 0.23 g) were distributed in six experimental groups in triplicates following the complete random distribution. Fish were fed twice daily with respective experimental diets for a period of 60 days. A significantly (p < .05) lower lactate dehydrogenase, malate dehydrogenase, superoxide dismutase and catalase activities were registered in supplemented groups compared with control group, while respiratory burst and lysozyme activities were significantly (p < .05) higher in E10 group compared with other experimental groups. Haemoglobin, total leucocyte count, total erythrocyte count and haematocrit values were significantly (p < .05) higher in E10 group. The expression of IFN‐γ and TNF‐α in both the kidney and liver was significantly up‐regulated in leaf extract and meal supplemented groups with the highest expression in the fish of E10 group. Overall, these results suggest that the dietary supplementation of ethanolic extract of the Houttuynia cordata leaf at 10 g/kg level can enhance the immune response of L. rohita fingerlings.     

Dietary phytase and protease improved growth and nutrient utilization in tilapia (Oreochromis niloticus × Oreochromis aureus) fed low phosphorus and fishmeal‐free diets

A 10‐week study was conducted to evaluate the effects of supplemental phytase and protease in low phosphorus (LP) and fishmeal‐free diets (FMF) on growth and nutrient utilization of tilapia. Seven isonitrogenous diets were prepared as positive control diet (PC) containing 60 g/kg fishmeal and 18 g/kg monocalcium phosphate (MCP), LP diet (12 g/kg MCP), FMF diet, LP‐FMF diet, and three enzyme diets with phytase (LP + Ph), protease (FMF + Pr), and phytase + protease (LP‐FMF + Ph + Pr) supplementation in LP, FMF and LP‐FMF diets, respectively. Weight gain, nutrient (except Ca) retention and digestibility of LP, FMF and LP‐FMF groups were lower than those of PC group (p < 0.05). Compared to LP, FMF and LP‐FMF groups, the supplementation of phytase, protease and phytase + protease improved WG by 13.2%, 11.1% and 13.0% (p < 0.05). The three enzyme groups showed higher digestibility of protein and P, and higher intestinal villus length than those of LP, FMF and LP‐FMF groups (p < 0.05), respectively. Dietary phytase and Ph + Pr also promoted the retentions of crude protein and P (p < 0.05). The results indicated that the supplementation of phytase in LP diet and protease in FMF diet improved the growth and nutrient utilization, and phytase reduced MCP inclusion in the diet of tilapia.