Effects of dietary protein and lipid levels on growth, feed conversion and body composition in rohu, Labeo rohita (Hamilton), fingerlings

Twelve experimental diets (D‐1 to D‐12) in a 4 × 3 factorial design (four protein levels: 250, 350, 400 and 450 g kg^?1 and three lipid levels: 50, 100 and 150 g kg^?1) were formulated. Carbohydrate level was constant at 250 g kg^?1. Rohu fingerlings (average wt. 4.3 ± 0.02 g) were fed the experimental diets for 60 days in three replicates at 2% BW  day^?1. Weight gain (%), specific growth rate (% day^?1) and feed gain ratio (FGR) indicated that diets containing 450 g kg^?1 protein and 100 or 150 g kg^?1 lipid (diets D‐11 and D‐12) resulted in best performance, although results were not significantly different from those of diet D‐9 (400 g kg^?1 protein and 150 g kg^?1 lipid). Protein efficiency ratio was highest with diets D‐6 (350 g kg^?1 protein and 150 g kg^?1 lipid) and D‐9 (400 g kg^?1 protein and 150 g kg^?1 lipid) (P > 0.05) and declined with higher and lower protein diets at all levels of lipid tested. Elevated lipid level (50, 100 or 150 g kg^?1) did not produce better FGR in diets containing 400 and 450 g kg^?1 dietary protein (P > 0.05). The combined effects of protein and lipid were evident up to 400 g kg^?1 protein. Growth and FGR showed consistent improvement with increased lipid levels from 50 to 150 g kg^?1 at each protein level tested except with diets containing 450 g kg^?1 protein. Apparent nutrient digestibility (for protein, lipid and energy) did not show significant variation among different dietary groups (P > 0.05). Whole body protein and lipid contents increased significantly (P > 0.05) with dietary protein level. The results of this study indicate that rohu fingerlings are adapted to utilize high protein in diets with varying efficiency. The fish could utilize lipid to spare protein but there is no significant advantage from this beyond the dietary protein level of 350–400 g kg^?1 in terms of growth and body composition.     

Effects of varying dietary protein levels on growth, reproductive performance, body and egg composition of rohu, Labeo rohita (Hamilton)

A 360‐day feeding trial was conducted to observe the influence of varying levels of dietary protein on growth, reproductive performance, body and egg composition of rohu, Labeo rohita. Twenty fish (40.4 ± 0.24 cm; 852 ± 4.9 g), stocked in outdoor concrete tanks (200 m²), in duplicate, were fed diets with varying levels (200, 250, 300, 350 and 400 g kg^?1) of crude protein exchanged with carbohydrate to apparent satiation, twice daily, at 09:00 and 17:00 h. Higher (P < 0.05) weight increment was discernible in fish fed dietary protein ≥300 g kg^?1. Gonadosomatic index was comparable (P > 0.05) among fish of different dietary groups except those fed 200 g kg^?1 protein diet which produced least values. Egg diameter remained unaffected (P > 0.05) by variations in levels of dietary protein. Relative fecundity was maximum (P < 0.05) in fish fed 250 and 300 g kg^?1 protein diets. With the exception of fish fed 200 g kg^?1 protein diet, fertilizability (%) remained unaffected (P > 0.05) by variations in dietary protein level. Hatchability (%) followed the trend of variations almost similar to that of fertilizability. Proximate composition of muscle and eggs varied significantly (P < 0.05) with dietary protein levels. For broodstock L. rohita, a dietary protein level of 250 g kg^?1 was found optimum with regard to its reproductive performance, egg quality and composition.     

Effect of dietary L-carnitine supplements on growth and body composition of fingerling rohu, Labeo rohita (Hamilton)

Rohu, Labeo rohita (Ham.), fingerlings (average wt 3.38 g) maintained in 25 m² cement tanks were fed fish-meal-based, 300 g kg⁻¹ protein diets containing different levels (0, 0.25, 0.5, 0.75 and 1.0 g kg⁻¹) of L -carnitine in triplicate over a period of 126 days. Fish that received 0.5 g kg⁻¹ carnitine showed significantly higher growth than those fed the basal diet or the rest of the carnitine diets. Fat digestibility, food conversion efficiency and protein efficiency ratio were better in carnitine-fed fish; however, viscerosomatic and hepatosomatic indices decreased. Muscle proximate composition was affected as a result of carnitine treatment. The results of the present study demonstrate a positive effect of carnitine on the growth and body composition of rohu.     

Allozyme variation of hatchery and river populations of rohu (Labeo rohita, Hamilton) in Bangladesh

The genetic variations of rohu (Labeo rohita, Hamilton) sampled from five hatchery populations (Arabpur, Brahmaputra, Comilla, Kishorganj and Natore) and three major river populations (the Halda, the Jamuna and the Padma) were analysed by allozyme electrophoresis. Ten enzymes encoded by 11 loci were screened, and six were polymorphic. Alleles at three loci (Est‐1^*, Gpi‐1^* and Gpi‐2^*) proved variable for hatchery and river populations, and the Mdh‐2^* locus exhibited heterozygous genotypes for river populations only. Polymorphic loci per population (27.3±5.3%), heterozygous loci per individual (15.5±1.2%) and relative gene diversity (0.27±0.08) in river populations were higher than those for hatchery populations (25.5±1.8%, 10.7±1.6% and 0.25±0.01 respectively). Also, the observed heterozygosity (Ho) and expected heterozygosity (He) (0.09±0.03 and 0.14±0.04 respectively) in river populations were higher than those in hatchery populations (0.08±0.01 and 0.11±0.01 respectively). The lower levels of genetic variability in hatchery populations suggested the occurrence of inbreeding and/or genetic drift. The pairwise population differentiation (F_ST) values showed a lower level of genetic differentiation between hatchery and river population pairs. The unweighted pair‐group method with arithmetic mean dendrogram of Nei's genetic distances showed a relationship between the genetic distance and geographic distance. The populations were clustered into three groups: the Padma in one group, the Halda in second group and the Jamuna, including five hatcheries, in the third group. Highly diversified rohu individuals were observed in the Padma and Halda Rivers, whereas less genetically variable individuals were found in the Jamuna River and five hatcheries. These findings can be useful for rohu hatchery propagation to enhance the sustainable aquaculture production.     

Total sulphur amino acid requirement and cystine replacement value for fingerling rohu,Labeo rohita: effects on growth,nutrient retention and body composition

Two feeding experiments were conducted to quantify the total sulphur amino acid (TSAA) requirement and replacement value of cystine for methionine for fingerling Labeo rohita. In Experiment I, isonitrogenous (380 g kg^?1 CP) and isocaloric (17.90 kJ g^?1 GE) amino acid test diets with graded levels of methionine (4, 6, 8, 10, 12, 14 g kg^?1 dry diet) and 0.4 g kg^?1 cystine were fed to fish (4.62 ± 0.2 cm; 0.66 ± 0.1 g) and methionine requirement determined by analysing absolute weight gain (AWG) (5.48), feed conversion ratio (FCR) (1.26), protein retention efficiency (PRE%) (39%) and energy retention efficiency (ERE%) (85%) data which were best at 10 g kg^?1 methionine of dry diet. In Experiment II, six diets with different ratios of L‐cystine and L‐methionine on equimolar sulphur basis were fed to fish (4.71 ± 0.1 cm; 0.69 ± 0.2 g) under identical conditions. Maximum AWG (5.58), best FCR (1.24), PRE (41%) and ERE (86%) in fish fed Diet IV indicated cystine replacement value to be 40%. On the basis of the broken‐line and second‐degree polynomial regression analyses of results obtained in Experiments I and II, it is concluded that inclusion of TSAA in the range of 25.2–31.31 g kg^?1 of protein is optimum of which 33–39% could be spared by cystine.     

The potential of periphyton-based culture of two Indian major carps, rohu Labeo rohita (Hamilton) and gonia Labeo gonius (Linnaeus)

The effects of periphyton, grown on bamboo substrates, on growth and production of two Indian major carps, rohu, Labeo rohita (Hamilton) and gonia, Labeo gonius (Linnaeus), were studied at the Bangladesh Agricultural University, Mymensingh. For each species, five ponds were provided with bamboo substrates and five ponds without substrate (control). Ponds were stocked at a rate of 10 000 ha^?1 in both treatments. There was no discernible difference in water quality parameters between treatments. A large number of plankton (39 genera) showed periphytic nature and colonized the bamboo substrates. Rohu grew faster, resulting in a 77% higher net production (P < 0.05) in the ponds with bamboo substrates compared with the ponds without substrate. In contrast, the growth and production of gonia did not vary significantly (P > 0.05) between the substrate and control ponds. Rohu seems to be a more suitable candidate for periphyton‐based aquaculture systems than gonia.     

Effects of dietary inclusion of the beta-adrenergic agonist, salbutamol, on growth performance and whole-body composition of rohu Labeo rohita (Hamilton) fingerlings fed diets containing two protein levels

A 60-day feeding experiment was conducted in the laboratory to evaluate the effect of oral administration of the beta-adrenergic agonist (BAA), salbutamol, on growth, nutrient utilization and whole-body composition of rohu Labeo rohita (Hamilton) fingerlings (average weight 5.51 ± 0.07 g) acting as a repartitioning agent in intermediary metabolism and redistributing nutrients for muscle synthesis. Two diets (diets 30/0 and 40/0) were formulated containing 30% and 40% crude protein to serve as basal diets containing the same ingredient composition. Another four diets (diets 30/3, 30/6, 40/3 and 40/6) were prepared in the same way to contain either 3 or 6 mg kg⁻¹ BAA salbutamol at each protein level making the total of six experimental diets. Rohu fingerlings were fed with the experimental diets in three replicate treatments at a restricted feeding regime equivalent to 2% of body weight (BW) day⁻¹. A significant (P < 0.05) interaction effect was found between dietary protein level and BAA treatment. Dietary incorporation of BAA at both protein levels significantly (P < 0.05) increased growth and nutrient utilization in terms of weight gain (%), specific growth rate (SGR), feed:gain ratio (FGR) and protein efficiency ratio (PER). BAA treatment at the 6 mg kg⁻¹ concentration led to a 12% rise in growth with the 30% protein diet and a 6% rise with the 40% protein diet, above their respective controls (without BAA treatment). However, no significant differences were found by raising the BAA concentration from 3 to 6 mg kg⁻¹. The apparent digestibility values for protein, lipid and energy were only higher with the 30% protein diets containing BAA. At both protein levels tested, BAA exerted a significant (P < 0.05) positive influence on protein retention and a negative influence on lipid retention efficiency, although the differences between the 3 and 6 mg kg⁻¹ BAA diets were not significant. This indicated that BAA acted as a repartitioning agent in suppressing lipid deposition in favour of protein accretion. BAA induced significantly (P < 0.05) higher whole-body protein and lipid relative to the untreated groups. The condition factor improved significantly (P < 0.05) in the BAA-treated dietary groups. The fillet percentage displayed a direct correlation and the frame percentage a negative correlation with BAA concentration in the diets. The results indicate that the BAA, salbutamol, was effective in producing growth enhancement, improved body composition (higher protein and lower lipid accretion) and efficient nutrient utilization at the 3 mg kg⁻¹ dietary incorporation level and thus has a potential for application in formulated diets for the Indian major carp, rohu, under culture conditions.     

Effects of paddy straw and sugarcane bagasse on water quality, bacterial biofilm production and growth and survival of rohu, Labeo rohita (Hamilton)

Sugarcane bagasse (T₁), paddy straw (T₂) and their mixture (T₃) were added to mud‐bottomed, manured cement tanks in triplicate to study the effect on bacterial biofilm production and growth and survival of rohu, Labeo rohita. All tanks were stocked with fingerling rohu at 10 000 ha^?1. The total plate count (TPC) of bacteria in water and on substrate did not differ significantly between treatments. The mean phytoplankton and zooplankton density in water was the highest in T₂, followed by T₃ and T₁. The growth of fish was significantly (P<0.05) higher in T₂ and T₃ compared with T₁. Significantly higher RNA values were recorded in T₂ and T₃ compared with T₁. Similarly, T₂ and T₃ registered higher DNA values than T₁. RNA:DNA ratio was higher in T₂ compared with T₁. However, T₃ did not differ significantly from T₁ and T₂. Both intestinal and hepatopancreatic protease and lipase activity was higher in T₁ and T₂ compared with T₃ and both differ significantly from T₁. Intestinal amylase activity was higher in T₂ and T₃ compared with T₁, whereas hepatopancreatic amylase activity was higher in T₂ compared with T₁ and T₃. The results demonstrated that production of L. rohita can be significantly increased by the introduction of biodegradable substrates into culture systems where fertilization is employed.     

Arginine and histidine requirements of the Indian major carp, Labeo rohita (Hamilton)

Growth studies were conducted to quantify requirements for the essential amino acids (EAA), arginine and histidine in Labeo rohita (Hamilton). Diets incorporating casein and gelatine, as sources of intact protein, supplemented with crystalline amino acids were formulated to a crude protein content of 400 g kg^?1. Diets with six graded levels of arginine (14.6, 17.0, 19.0, 21.0, 23.0 and 25 g kg^?1) and histidine (3.2, 5.0, 7.0, 9.0, 11.0 and 13.0 g kg^?1) were formulated and fed to triplicate groups of juvenile rohu twice a day up to satiation for 60 days. Dietary requirements for arginine and histidine for rohu, estimated using break point analysis, were 23 and 9 of the diet respectively (57.5 and 22.5 g kg^?1 of dietary protein). Food conversion rate, specific growth rate and survival were better in treatments with diets containing optimum levels of EAA.     

Total sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton)

The quantitative dietary sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton), was determined by conducting a growth study. The experimental diets contained 400 g crude protein kg⁻¹ from casein, gelatine and supplemental crystalline amino acids. Diets containing six graded levels of methionine (3.2, 6.5, 9.0, 11.5, 14 and 16.5 g kg⁻¹) with a constant level of cystine (1.4 g kg⁻¹) were formulated and fed to triplicate groups of Labeo rohita fingerlings twice a day to satiation for 60 days. The optimum dietary requirement for methionine was estimated using the break-point regression analysis at 11.5 g kg⁻¹ of diet or 28.8 g kg⁻¹ of dietary protein. Thus the total sulphur amino acid (Met + Cys) requirement was determined to be 12.9 g kg⁻¹ of diet or 32.3 g kg⁻¹ of protein. Higher survival, specific growth rate and food conversion efficiency values were observed for fish fed the diet containing optimum levels of sulphur amino acids.     

Dietary protein level, microbial phytase, citric acid and their interactions on bone mineralization of Labeo rohita (Hamilton) juveniles

A feeding trial was conducted for 60 days to study the effect of dietary protein, microbial phytase and citric acid on intestinal digesta pH, bone ash and bone mineral contents in Labeo rohita juveniles. Eight experimental diets were prepared in 2 × 2 × 2 factorial arrangement with crude protein levels (25% and 35%), microbial phytase (0 and 500 U kg^?1), and citric acid (0 and 3%). The 25% crude protein level feed was supplemented with phytase (U kg^?1) and citric acid (%) at the level of 0,0 (C₂₅); 500,0 (T₁); 0,3 (T₂); 500,3 (T₃), and 35% crude protein level feed at 0.0 (C₃₅); 500,0 (T₄); 0,3 (T₅) and 500,3(T₆) respectively. One hundred and twenty juveniles of L. rohita (av. wt. 12.61–13.72 g) were distributed randomly in eight treatments, each of with three replicates. Addition of citric acid in the 25% crude protein feed significantly decreased (P<0.001) feed pH with concurrent decrease in intestinal digesta pH (P<0.001) and increased the bone ash content (P<0.05) by 4.6%. An interaction between citric acid and phytase (P<0.05) was also observed for bone ash content. Increasing the dietary protein content from 25% to 35% significantly decreased (P<0.01) bone Zn content by 14.9%, which was more prominent with the addition of citric acid, resulting in significant interaction between protein and citric acid (P<0.05), but the bone Cu content was significantly increased (P<0.01) with increasing dietary protein content. Dietary supplementation of microbial phytase (500 U kg^?1) significantly increased (P<0.05) bone Na, Ca, K, P and Fe contents by 15%, 12.1%, 17.4%, 9.2% and 40.7%, respectively, whereas bone P and Mn content was significantly increased (P<0.05) by addition of citric acid (3%). Addition of phytase to plant‐based diets increased the bioavailability of minerals, thereby increasing bone mineralization. The effect of phytase was increased because of addition of citric acid (3%).     

Dietary isoleucine requirement of fingerling Indian major carp, Labeo rohita (Hamilton)

An 8‐week feeding experiment was conducted to quantify the dietary isoleucine requirement of fingerling Indian major carp, Labeo rohita (3.50 ± 0.04 cm; 0.40 ± 0.02 g) using amino acid test diets (400 g kg⁻¹ crude protein; 17.90 kJ g⁻¹ gross energy) containing casein, gelatin and l ‐crystalline amino acids. Six dietary treatments supplemented with graded levels of isoleucine (7.5, 10.0, 12.5, 15.0, 17.5 and 20.0 g kg⁻¹), in gradations of 2.5 g kg⁻¹ diet, were fed to triplicate groups of fingerlings to apparent satiation divided over two feedings at 07:00 and 17:30 h. Performance of the fish was evaluated on the basis of live weight gain, feed conversion ratio (FCR), protein efficiency ratio (PER), specific growth rate (SGR) and protein productive value (PPV). Statistical analysis of live weight gain, FCR, PER, SGR and PPV reflected significant differences among treatments. Live weight gain and conversion efficiencies were best with isoleucine at 15.0 g kg⁻¹ of diet. Live weight gain, FCR, PER, SGR and PPV data were also analysed using second‐degree polynomial regression analysis to obtain more accurate isoleucine requirement estimate which was found to be at 15.9, 15.3, 15.2, 15.8 and 15.7 g kg⁻¹ of dry diet, corresponding to 39.8, 38.3, 38.0, 39.5 and 39.3 g kg⁻¹ of dietary protein respectively. Based on the quadratic regression analysis of the live weight gain, FCR, PER, SGR and PPV, the optimum level of isoleucine for fingerling L. rohita is in the range of 15.2–15.9 g kg⁻¹ of dry diet, corresponding to 38.0–39.8 g kg⁻¹ of dietary protein. Maximum body protein, minimum moisture and fat were noted at 15.0 g kg⁻¹ of dietary isoleucine while the body ash remained constant among all the treatment levels. No mortality was recorded during the duration of the experiment.     

Growth Response and Biochemical Composition of Rohu,Labeo rohita,Fed Salt-Incorporated Diets

Abstract

Rohu, Labeo rohita, fingerlings (av. wt. 1.6 g) were fed pelleted diets supplemented with 0,0.5,1.0,1.5 and 2.0% sodium chloride (NaCl) for 120 days in 25-m³ cement tanks. The final weight gain of fish fed 0.5 and 1.0% NaCl-incorporated diets was significantly (P < 0.05) higher than that of the control (0% NaCl). Net protein retention and carcass protein and fat contents were higher under different treatments as compared to the control. Growth indices and carcass composition indicate inclusion of 1.0% NaCl in the diet to be beneficial for growth promotion in rohu.     

Dietary leucine requirement of fingerling Indian major carp, Labeo rohita (Hamilton)

An 8-week feeding experiment was conducted to evaluate the dietary leucine requirement of fingerling Indian major carp, Labeo rohita (3.50±0.04 cm; 0.40±0.02 g) using amino acid test diets (40% crude protein; 17.90 kJ g⁻¹ gross energy) containing casein and gelatin as intact protein sources and l -crystalline amino acids. Growth performance and biochemical parameters were assessed by feeding six amino acid test diets supplemented with graded concentrations of leucine (0.75, 1.0, 1.25, 1.50, 1.75 and 2.0 g per 100 g) to triplicate groups of fingerlings to apparent satiation divided over two feedings at 07:00 and 17:30 hours. Performance of the fish was evaluated on the basis of live weight gain, feed conversion ratio (FCR), protein efficiency ratio (PER) and body protein deposition (BPD) data. Maximum live weight gain (315%), best FCR (1.35), highest PER (1.86) and BPD (33.9) were recorded at 1.50 g per 100 g dietary leucine. Statistical analysis of live weight gain, FCR, PER and BPD data reflected significant differences (P<0.05) among treatments. Live weight gain, FCR, PER and BPD data were also analysed using second-degree polynomial regression analysis to obtain more accurate leucine requirement estimate which was found to be at 1.57, 1.55, 1.52 and 1.50 g per 100 g of dry diet, corresponding to 3.92, 3.87, 3.80 and 3.75 g per 100 g of dietary protein respectively. Based on the quadratic regression analysis of the live weight gain, FCR, PER and BPD data, the optimum requirement of fingerling L. rohita for leucine is estimated to be in the range of 1.50–1.57 g per 100 g of the dry diet, corresponding to 3.75–3.92 g per 100 g of dietary protein.     

Dietary threonine requirement of fingerling Indian major carp,Labeo rohita (Hamilton)

An 8‐week feeding experiment was conducted in a water flow‐through system (26–28 °C) to determine the dietary threonine requirement of fingerling Labeo rohita (3.90±0.03 cm; 0.58±0.02 g). Growth, feed utilization and body composition of fish fed test diets (40% crude protein; 17.9 kJ g^?1 gross energy) with graded levels of l ‐threonine (0.75%, 1.0%, 1.25%, 1.50%, 1.75% and 2.0% dry diet) to apparent satiation were response variables used to assess threonine adequacy. Diets were made isonitrogenous and isoenergetic by adjusting the levels of glycine and dextrin. The amino acid profiles of the test diets were formulated to that of 40% whole chicken egg protein except for threonine. The performance of fish fed experimental diets was evaluated using calculated values for weight gain (g fish^?1), feed conversion ratio (FCR), protein efficiency ratio (PER) and protein productive value (PPV) data. Maximum weight gain (g fish^?1) (1.79), lowest FCR (1.39), highest PER (1.76) and PPV (0.33) were recorded at 1.50 g per 100 g dietary threonine. Statistical analysis of weight gain, FCR, PER and PPV data reflected significant differences (P<0.05) among treatments. Except for reduced growth performance in fish fed threonine‐deficient diets, no deficiency signs were noted. Weight gain, FCR, PER and PPV data were also analysed using second‐degree polynomial regression analysis to obtain a more accurate threonine requirement estimate, which was found, using each response variable, to be at 1.70, 1.63, 1.65 and 1.51 g per 100 g of dry diet, corresponding to 4.2, 4.07, 4.12 and 3.77 g per 100 g of dietary protein respectively. Based on the second‐degree polynomial regression analysis of the live weight gain, FCR, PER and PPV data, the optimum dietary level of threonine for fingerling L. rohita was found to be in the range of 1.51–1.70 g per 100 g of the dry diet, corresponding to 3.77–4.2 g per 100 g of dietary protein.     

Dietary Phosphorus Requirement of Fingerling Indian Major Carp,Labeo rohita (Hamilton)

Dietary phosphorus requirement of fingerling Labeo rohita (6.1 ± 0.13 cm; 1.88 ± 0.05 g) was quantified by feeding seven isonitrogenous (350 g/kg crude protein) and isocaloric (16.72 kJ/g gross energy) purified diets with different levels of phosphorus as 3.5 (basal diet), 4.6, 5.7, 6.5, 7.8, 8.9, and 10.1 g/kg. Triplicate groups of fish were fed at 0800, 1200, and 1600 h to apparent satiation for 8 wk. Live weight gain (LWG; 494.68%), specific growth rate (3.18%/d), feed conversion ratio (1.54), feed efficiency (0.65), protein gain (PG; 1.26 g/fish), protein efficiency ratio (1.86), and phosphorus utilization efficiency (98.78%) improved significantly (P < 0.05), with increasing dietary phosphorus level up to 6.5 g/kg. However, phosphorus contents of vertebrae and scale increased significantly up to 7.8 g/kg. Dietary phosphorus levels significantly affected serum phosphorus concentration and alkaline phosphatase activity. Broken‐line analysis based on LWG; PG; and whole‐body, vertebrae, and scale phosphorus against dietary phosphorus indicated the optimal phosphorus requirement of fingerling L. rohita at 6.56, 6.58, 6.56, 8.02, and 8.44 g/kg diet, respectively. In order to restrict superfluous phosphorus in the diet, inclusion of 6.56 g/kg phosphorus is recommended for optimal growth of fingerling L. rohita.     

Development and characterization of cell lines derived from rohu, Labeo rohita (Hamilton), and catla, Catla catla (Hamilton)

Two new cell lines, designated RE and CB, were derived from the eye of rohu, Labeo rohita , and the brain of catla, Catla catla , respectively. The cell lines were maintained in Leibovitz's L-15 supplemented with 20% foetal bovine serum. The RE cell line was sub-cultured for more than 70 passages and the CB cell line for more than 35 passages. The RE cells are rounded and consist predominantly of epithelial cells. The CB cell line consists of predominantly fibroblastic-like cells. Both cell lines are able to grow at temperatures between 25 and 32 °C with an optimum of 28 °C. The growth rate of the cells increased as the foetal bovine serum concentration increased from 2% to 20% at 28 °C, with optimum growth at concentrations of 15% or 20% foetal bovine serum. The cells were successfully cryopreserved and revived at different passage levels. The cell lines were not susceptible to four marine fish viruses. Extracellular products from Aeromonas sp . were toxic to the cell lines. When the cells were transfected with plasmid eukaryotic green fluorescent protein (pEGFP [Clontech, Carlsbad, CA, USA]) vector DNA, a significant fluorescent signal was observed suggesting that these cell lines could be a useful tool for transgenic and genetic manipulation studies. Polymerase chain reaction amplification of mitochondrial 12S rRNA from rohu and catla confirmed that the cell lines originated from these fish species. The cell lines were further characterized by immunocytochemistry using confocal laser scanning microscopy.     

Dietary microbial phytase and citric acid synergistically enhances nutrient digestibility and growth performance of Labeo rohita (Hamilton) juveniles at sub-optimal protein level

A 2 × 2 × 2 factorial experiment was designed to study the main effect of dietary microbial phytase, citric acid, crude protein (CP) level and their interactions on growth performance, nutrient digestibility and body composition of Labeo rohita juveniles. Two basal diets were formulated using plant‐based ingredients containing either sub‐optimum (25%) or normal (35%) CP levels. Both the diets were supplemented with microbial phytase (U kg⁻¹) and citric acid (%) at the level of 0, 0; 500, 0; 0, 3; and 500, 3 respectively. One hundred and twenty L. rohita juveniles (average weight 12.61–13.72 g) were randomly distributed in eight treatments, each with three replicates. Dietary microbial phytase alone had no significant effect on whole‐body ash content but addition of citric acid (3%) in the diet activated the phytase as evidenced by their significant interaction. A significant interaction between citric acid and microbial phytase was also found on weight gain%, specific growth rate, protein efficiency ratio. Increasing the dietary CP level from 25% to 35% significantly (P<0.01) decreased phosphorus and dry matter digestibility. Thus, these results showed that the growth‐promoting effect was higher in groups fed a sub‐optimum protein (25%) diet containing both 3% citric acid and 500 U kg⁻¹ of microbial phytase than those fed 35% CP diets. This suggests that microbial phytase and citric acid in sub‐optimum CP diet had a synergistic effect on nutrient digestibility and growth performance of L. rohita juveniles.     

Stress mitigating and immunomodulatory effect of dietary pyridoxine in Labeo rohita (Hamilton) fingerlings

A 60‐day experiment was carried out to delineate stress mitigating and immunomodulatory role of dietary pyridoxine (PN) in Labeo rohita fingerlings exposed to endosulfan. Two hundred and seventy fingerlings were randomly distributed into six treatments in triplicates. Five iso‐caloric and iso‐nitrogenous purified diets were prepared with graded levels of pyridoxine. Six treatment groups were T₀ (10 mg PN+without endosulfan), T₁ (0 mg PN+endosulfan), T₂ (10 mg PN+endosulfan), T₃ (50 mg PN+endosulfan), T₄ (100 mg PN+endosulfan) and T₅ (200 mg PN+endosulfan). The role of pyridoxine on stress mitigation and immunomodulation was assessed by biochemical and haemato‐immunological parameters like aspartate aminotransaminase, alanine aminotransaminase, lactate dehydrogenase, malate dehydrogenase, superoxide dismutase and catalase were significantly (P<0.05) lower while acetylcholinesterase was significantly (P<0.05) higher in pyridoxine‐fed groups. Erythrocytes count, haemoglobin content and total serum protein, albumin, globulin, nitroblue tetrazolium and lysozyme activity were significantly (P<0.05) higher while cortisol and blood glucose were decreased significantly (P<0.05) in pyridoxine‐fed groups. Percentage survival after challenge with Aeromonas hydrophila was highest in T₀ group. The results obtained in present study indicate that dietary pyridoxine supplementation at 100 mg PN kg^?1 diet reduces the endosulfan‐induced stress and triggers immune response in L. rohita fingerlings.     

Characteristics of Collagen from Rohu (Labeo rohita) Skin

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from rohu skin with the yield of 64.2 and 6.8% (dry weight basis), respectively. Both collagens had glycine as the major amino acid with imino acid content of 196–202 residues/1,000 residues and were characterized as type I collagen with molecular composition of (α1)₂α2-heterotrimer. Fourier transform infrared spectra of both collagens were similar, with no shift in wavenumber of all amide bands. The T_max value of ASC and PSC was 36.40 and 35.48°C, respectively. The zero surface net charge of ASC and PSC was found at pH 5.9 and 5.3, respectively.