Effect of various sources of dietary additive on growth,body composition and serum chemistry of juvenile olive flounder (Paralichthys olivaceus)

Asia traditional herbal feed additives have often been shown to have beneficial effects on disease resistance, growth, feed conversion and protein conversion. There have been few studies on the use of these feed additives in olive flounder diets. The present study was designed to determine if there are benefits associated with the inclusion of any of these traditional herbal feed additives in olive flounder diets. One thousand two hundred sixty juvenile fish averaging 6.5 g were randomly distributed into 36, 180‐L flow‐through tanks. No additive was included into the control (Con) diet. The aqueous type of additives: 2.5% bamboo (BB), 0.25% Prunus mume (PM), 2.5% Artemisia capillaris (AC), 2.5% cinnamon (CC), 2.5% Cirtus junos (CJ), 0.25% Cudrania tricuspidata (CT) and 2.5% Ficus carica (FC), and the powder type of additives: 1% Houttuynia cordata (HC), 1% safflower (SF) and 1% sea tangle (ST) were included into the experimental diets. Finally, 0.1% emulsifying oil (EO) was included into the experimental diet. Feed efficiency ratio (FER) of fish fed the BB, CT and SF diets was higher than that of fish fed the FC and HC diets. Protein efficiency ratio (PER) of fish fed the BB and SF diets was higher than that of fish fed the AC, FC and HC diets. None of serum criteria of fish was affected by any of dietary additives. Dietary additives did not affect weight gain and serum chemistry of olive flounder, but 2.5% bamboo extract and 1% safflower powder was effective to improve FER and PER.     

Optimum dietary protein level for maximum growth of juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel)

This experiment was conducted to determine the optimum dietary protein level for juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel) fed a white fish meal and casein‐based diets for 8 weeks. Olive flounder with an initial body weight of 4.1 ± 0.02 g (mean ± SD) were fed one of the six isocaloric diets containing 35%, 45%, 50%, 55% and 65% crude protein (CP) at a feeding rate of 4–5% of wet body weight on a dry‐matter basis to triplicate groups of 20 fish per aquarium. After 8 weeks of feeding, per cent weight gain (WG) and feed efficiency ratios of fish fed the 55% CP diet were not significantly higher than those from fish fed the 50% and 65% CP diets, but significantly higher than those from fish fed the 35% and 45% CP diets. Fish fed the 50%, 55% and 65% CP diets had significant higher specific growth rates than did fish fed the 35% and 45% CP diets; however, there was no significant difference among fish fed the 50%, 55% and 65% CP diets. The protein efficiency ratio was inversely related to the dietary protein level; that is, maximum efficiency occurred at the lowest dietary protein level. Broken‐line model analysis indicated that the optimum dietary protein level was 51.2 ± 1.8% for maximum weight gain in juvenile olive flounder. The second‐order polynomial regression analysis showed that the maximum WG occurred at 57.7% and it revealed that the minimum range of protein requirement was between 44.2% and 46.4%. These findings suggest that the optimum dietary protein level for maximum growth could be greater than 46.4%, but less than 51.2% CP in fish meal and casein‐based diets containing 17.0 kJ g^?1 energy for juvenile olive flounder.     

Effect of dietary nutrient composition on compensatory growth of juvenile olive flounder Paralichthys olivaceus using different feeding regimes

Effect of dietary nutrient composition on compensatory growth of juvenile olive flounder, Paralichthys olivaceus using different feeding regimes was determined. Six treatments were prepared in triplicate: fish were daily hand‐fed with the control (C) diet twice a day, 7 days a week, for 8 weeks (8WF‐C); fish were starved for 1 week, and then fed with the C and high protein and lipid (HPL) diets twice a day, 7 days a week, for 7 weeks, referred to as 7WF‐C and 7WF‐HPL, respectively; and fish were starved for 2 weeks, and then fed with the C, HPL and HPL with supplementation of the mixture of amino acids (HPLA) diets twice a day, 7 days a week, for 6 weeks, referred to as 6WF‐C, 6WF‐HPL and 6WF‐HPLA, respectively. Growth of fish in 7WF‐HPL treatment was greater than that of fish in any other treatments. Feed efficiency of fish in 7WF‐HPL, 6WF‐HPL and 6WF‐HPLA treatments was higher than that of fish in 8WF‐C, 7WF‐C and 6WF‐C treatments. The high protein and lipid diet effectively improved compensatory growth of fish. However, supplementation of the mixture of amino acids into the high protein and lipid diet did not improve compensatory growth of fish further.     

Dietary choline requirement of juvenile olive flounder (Paralichthys olivaceus)

An eight‐week study was conducted to determine the optimum dietary choline level in juvenile olive flounder, Paralichthys olivaceus. Seven diets were prepared to contain 0, 250, 500, 750, 1,000, 2,000 and 3,000 mg/kg diet. Juveniles (5.9 ± 0.03 g; 5.5 ± 0.4 cm; mean ± SD) were randomly distributed into 21 tanks (25 fish/tank) and fed one of the diets in triplicates. Survival rate of fish fed the diet containing the lowest choline level was significantly lower than those of fish fed the other diets (p < 0.05). Final body weight, weight gain, specific growth rate, feed efficiency and protein efficiency ratio significantly increased with increasing choline levels up to 1,000 mg/kg diet. Whole‐body protein and lipid contents increased in accordance with choline levels up to 750 mg/kg diet, beyond which they plateaued. Liver and muscle lipid contents elevated with increasing choline levels up to 2,000 mg/kg diet. Plasma cholesterol esters, triglycerides, cholesterol and total lipids were significantly influenced by the graded choline levels; however, responses of those indices were not identical. Broken‐line analyses of weight gain and liver choline concentrations responding to the graded choline levels revealed that choline requirements of the juvenile flounder could be between 847 and 1,047 mg/kg diet.     

Effects of feeding frequency and feed type on the growth,feed utilization and body composition of juvenile olive flounder,Paralichthys olivaceus

A factorial (3 × 4) feeding trial was conducted to investigate the effects of feeding frequency and feed type on the growth performance, feed utilization and body proximate composition of juvenile olive flounder. Three feed types including a sinking moist pellet (MP), a sinking dry pellet (DP) and a floating extruded pellet (EP) were used. Fish (average weight, 11.1 ± 0.4 g) were randomly distributed in thirty‐six 300 L tanks in a flowthrough system. Triplicate groups (30 fish per tank) of fish were fed each feed type to visual satiation at three meals per day, two meals per day, one meal per day and one meal every 2 days for 7 weeks. At the end of the feeding trial, the survival of fish was not significantly different among the treatments. Weight gain was affected by the feeding frequency. The highest weight gain was observed in fish fed the EP at three meals per day. The daily feed intake of fish fed the MP and DP tended to be higher than that of fish fed the EP at each feeding frequency. The feed efficiency and the protein efficiency ratio of fish fed the EP were higher than those of fish fed the MP and DP. The body proximate composition of fish was altered by the feeding frequency. The crude lipid content increased with the increase in the feeding frequency. The present findings suggest that three feedings per day at visual satiation of the floating EP may be sufficient for the maximal growth performance of olive flounder grown from 11 to 60 g.     

Optimal dietary taurine level for growth of juvenile Japanese flounder Paralichthys olivaceus

The requirement for taurine in juvenile Japanese flounder Paralichthys olivaceus was determined by feeding diets containing various levels of taurine and cystine. Test diets supplemented with 0.5, 1.0 and 1.5% of taurine or with 0.5, 1.0 and 1.5% of L -cystine were prepared. The basal diet contained 55% protein from white fish meal. These diets were fed to juvenile Japanese flounder with an initial mean bodyweight of 0.9 g (total length (TL) 48 mm) for 5 weeks. Approximately 1.4% taurine content in the diet was required for optimum growth of juvenile flounder. A positive linear relationship was noted between the content of taurine accumulated in the muscle, liver and brain and the level of taurine in the diet. However, there was no increased taurine content in tissues of fish fed the cystine-supplemented diet. In contrast, the fish fed control and cystine-supplemented diets showed higher contents of cystathionine in the tissues. The concentration of cystathionine in tissues rapidly decreased with an increase of taurine in the diet. It was also observed that for each of the dietary groups, a trace amount of taurine was excreted. These results suggest that the taurine content in the diet affects the sulfur amino acid metabolism of juvenile Japanese flounder, and indicate that juvenile flounder are unable to biosynthesize taurine from cystine.     

Effects of dietary arginine and lysine levels on growth performance and biochemical parameters of juvenile Japanese flounder Paralichthys olivaceus

ABSTRACT: To investigate the effects of disproportionate levels of dietary arginine and lysine on juvenile Japanese flounder Paralichthys olivaceus , growth performance and biochemical parameters were evaluated by feeding five test diets, comprising different levels of arginine and lysine, to triplicate groups of juveniles (initial bodyweight 1.85 g) for 40 days. Crystalline amino acids were supplemented to test diets to correspond to the amino acid pattern found in the whole body protein of the Japanese flounder, except for arginine and lysine. After the feeding trials, plasma arginine, lysine and urea levels, excreted ammonia-N, and liver arginase activity were analysed. Survival, specific growth rate, feed conversion efficiency, and apparent protein retention were adversely affected ( P  <   0.05) by the deficiency in dietary arginine or lysine concentrations. An excess of either lysine or arginine in the diet did not depress growth when the diets contained adequate levels of either arginine or lysine, proving that there is no evidence for an arginine–lysine antagonism. Results for plasma arginine, lysine and urea levels, excreted ammonia, and liver arginase activity also demonstrated that Japanese flounder juveniles are not sensitive to excess dietary levels of lysine and arginine.     

Effects of different dietary levels of L-ascorbyl-2-polyphosphate on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel)

This experiment was conducted to study the effects of different dietary levels of vitamin C, L‐ascorbyl‐2‐polyphosphate (ASPP), on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel). Fish were fed one of six semi‐purified diets containing an equivalent of 0, 25, 50, 75, 150, or 1500 mg ascorbic acid (AA) kg^?1 diet (C₀, C₂₅, C₅₀, C₇₅, C₁₅₀ or C₁₅₀₀) in the form of ASPP for 12 weeks. Weight gain (WG) and protein efficiency ratio (PER) of fish fed the C₀ diet were significantly lower than those of fish fed the other diets (P < 0.05), and WG and PER of fish fed the C₂₅, C₅₀ and C₇₅ diets were significantly lower than those of fish fed the C₁₅₀₀ diet (P < 0.05). Fish fed the C₀ diet exhibited vitamin C deficiency symptoms such as anorexia, scoliosis, cataract, exophthalmia and fin hemorrhage at the end of the 12‐week test. After 12 weeks of the feeding trial, AA concentrations from gill, kidney, and liver of fish fed the C₀, C₂₅, C₅₀ and C₇₅ diets were significantly lower than those of fish fed the C₁₅₀ and C₁₅₀₀ diets (P < 0.05). Based on broken line analyses for WG and PER, the optimum dietary levels of vitamin C were 91 and 93 mg AA kg^?1 diet respectively. These findings suggest that the dietary vitamin C requirement could be 93 mg AA kg^?1 diet to support reasonable growth, and greater than 150 mg AA kg^?1 diet may be required for AA saturation of major tissues for juvenile olive flounder under experimental conditions.     

Effects of feeding frequency and dietary energy level on growth and body composition of juvenile flounder, Paralichthys olivaceus (Temminck & Schlegel)

A two (dietary energy levels; low energy, LE and high energy, HE) × 4 (feeding frequency; one feeding in 2 days, one feeding daily, two times daily and three times daily) factorial experiment was performed to determine the effects of dietary energy level and/or feeding frequency on the growth and body composition of juvenile flounder Paralichthys olivaceus. The survival rate was not significantly different among treatments. Weight gain of fish fed the LE and HE diets significantly increased as feeding frequency increased. The weight gain of fish fed the LE diet was higher at each level of increasing feeding frequency. The weight gain of fish fed the HE diet was higher than that of fish fed the LE diet once in 2 days and once daily, but weight gain of fish fed the HE diet was lower than the LE diet three times daily. Daily feed intake was significantly influenced by feeding frequency, but not by dietary energy level. Feed efficiency of fish fed the HE diet once daily was significantly higher than that of fish fed the LE diet once in 2 days, but no significant difference in feed efficiency was observed among other groups of fish. The protein efficiency ratio of fish fed the LE diet once in 2 days was the lowest. Feeding frequency and dietary energy level had a significant effect on the body lipid content. A feeding frequency of two times or three times daily would be effective, depending on dietary energy level for maximum growth of juvenile flounder grown from 3.5 to 15 g.     

Muscle fiber growth in olive flounder,Paralichthys olivaceus: Fiber hyperplasia at a specific body weight period and continuous hypertrophy

The objective of this study was to investigate muscle fiber growth‐related attributes, especially fiber hyperplasia and hypertrophy, and their relation to growth characteristics in olive flounder. After a conditioning period, juvenile olive flounder (214 g of initial body weight) were weighed after 0, 4, 8, 12, and 24 weeks of a feeding trial and were selected based on mean body weight at each period (G1, G2, G3, G4, and G5, respectively). During the 3.0‐fold increase in body weight (363.1–1,071 g, p < 0.001), the fiber area was increased approximately 2.2 times (p < 0.05), and the median area increased from the G1 to G5 groups (p < 0.05). The G3 group exhibited a greater fiber number than the G2 group (p < 0.05) as the G3 group exhibited a higher percentage of smaller fiber and mosaic hyperplasia. In addition, a higher percentage of centered nuclei and deoxyribonucleic acid amount in the G2 group were accompanied by an increase in fiber number in the G3 group. Therefore, the muscle growth of olive flounder was influenced by continued fiber area growth and a significant number increase by mosaic hyperplasia during a specific period (G2 to G3 groups; from 512.7 to 670.3 g).     

Effect of dietary hydrolysate supplementation on growth performance,non‐specific immune response and disease resistance of olive flounder (Paralichthys olivaceus) challenged with Edwardsiella tarda

A 9‐week feeding trial was conducted to investigate the effects of dietary supplementation with protein hydrolysates on growth, innate immunity and disease resistance of olive flounder. A fishmeal (FM)‐based diet was regarded as a control, and three diets were prepared by partial replacement of FM with krill hydrolysate, shrimp hydrolysate or tilapia hydrolysate (designated as Con, KH, SH and TH, respectively). Triplicate groups of fish (24.5 ± 0.3 g) were fed one of the diets to apparent satiation twice daily for 9 weeks and then challenged by Edwarsellia tarda. Fish‐fed KH diet showed significantly (P < 0.05) higher growth performance and feed utilization compared with the Con diet. Dry matter digestibility of the diets was significantly increased by KH and TH supplementation. All the examined innate immune responses were significantly increased in fish fed KH diet. Significantly, higher respiratory burst and superoxide dismutase (SOD) activities were found in fish‐fed SH diet. Lysozyme and SOD activities were significantly increased in fish‐fed TH diet. However, no significant effect was found on fish disease resistance. This study indicates that dietary supplementation of the hydrolysates, particularly KH, can improve growth performance, feed utilization and innate immunity of olive flounder.     

Synergistic effects of dietary vitamin E and selenomethionine on growth performance and tissue methylmercury accumulation on mercury‐induced toxicity in juvenile olive flounder,Paralichthys olivaceus (Temminck et Schlegel)

An 8‐week feeding trial was conducted to evaluate the synergistic effects of dietary vitamin E and selenomethionine (SeMet) on induced methylmercury (MeHg) toxicity in juvenile olive flounder Paralichthys olivaceus. Nine semi‐purified diets were formulated to contain three different vitamin E levels as DL‐α‐tocopheryl acetate (0, 100 and 200 mg TAkg^?1 diet) and three different selenium (Se) levels (0, 2 and 4 SeMet mg kg^?1 diet) on the constant mercury toxicity level (20 mg MeHgkg^?1 diet). Nine experimental diets, in a 3² factorial design (E₀Se₀, E₀Se₂, E₀Se₄, E₁₀₀Se₀, E₁₀₀Se₂, E₁₀₀Se₄, E₂₀₀Se₀, E₂₀₀Se₂ and E₂₀₀Se₄), were fed to triplicate groups of fish averaging 2.3 ± 0.04 g (mean ± SD) in the semi‐recirculation system. After 8 weeks of feeding trial, vitamin E and Se showed significant effects on weight gain (WG) of fish (P < 0.05). We found that there was a clear trend of increasing WG with elevating vitamin E and Se levels in the diets. Feed efficiency (FE), specific growth rate (SGR), protein efficiency ratio (PER) and survivability exhibited a similar trend with WG. Both antioxidants had significant interaction effects on FE and PER (P < 0.05). Methylmercury concentrations in fish muscle, liver and kidney decreases in a dose‐dependent manner as dietary vitamin E and Se levels increase. Interestingly, the most significant interactive effects of vitamin E and Se were found in liver tissue for depleting Hg concentrations (P < 0.05). These findings suggest that dietary vitamin E more than 100 mg TA kg^?1 diet with 2 or 4 mg SeMet kg^?1‐supplemented diets could have synergistic effects on growth and liver mercury bioaccumulation on MeHg‐induced toxicity in juvenile olive flounder.     

Expression of insulin-like growth factor genes in olive flounder,Paralichthys olivaceus,fed a diet with partial replacement of dietary fish meal

This study analyzed the expression of growth-related genes of olive flounder, Paralichthys olivaceus, fed a low-fish meal (FM) diet to investigate the replacement of the FM diet in the field. The expression of growth hormone (GH)/insulin-like growth factor (IGF) axis genes in the brain/pituitary/liver and plasma hormone concentrations were measured. A basal experimental diet was formulated using 63% FM and 12% soybean meal as the primary protein sources, and then two other diets were made by replacing 20% and 30% of the FM with soy protein concentrate, tankage meal, and poultry by-products meal. Each diet was fed to duplicate groups of juvenile flounder (150 ± 3.0 g) twice a day. After 20 weeks, the plasma growth-related hormone concentrations were similar between the control and FM20 groups. Moreover, GH/IGF axis gene mRNA expression in the brain/pituitary/liver was similar between the control and FM20 groups. Immunoblotting of muscle and liver showed the same pattern. This study showed the utility of replacing 20% of the FM diet and supports the possibility of field application.     

Supplemental iron and phosphorus increase dietary inclusion of cottonseed and soybean meal in olive flounder (Paralichthys olivaceus)

A long‐term feeding experiment was conducted to investigate the use of cottonseed and soybean meal (CS) with iron and phosphorus supplements in diets for olive flounder (Paralichthys olivaceus). Olive flounder with an initial average size of 28.5 ± 0.35 g (mean ± SD) were divided into 15 groups (three tanks per dietary treatment) and fed 480 g kg^?1 crude protein diets in which each of five isonitrogenous diets was formulated to contain different levels of cottonseed/soybean meal (1 : 1) to replace fish meal (FM) with iron and phosphorus supplementations. The five experimental diets were as follows: diet 1 (control), 0%CS; diet 2, 20%CS; diet 3, 30%CS; diet 4, 30%CS + Fe&P; and diet 5, 40%CS + Fe&P. After 26 weeks of feeding trial, no significant differences were observed in weight gain, feed utilization and survival among all the treatments. The total gossypol accumulation in liver of fish fed diets supplemented with iron was significantly lower than that of fish fed diets without supplementation of iron. The results indicate that the addition of iron in diets could prevent the absorption of free gossypol. The findings in this study suggest that dietary supplements of iron and phosphorus could increase the inclusion of cottonseed and soybean meal for FM replacement in diets for marine fish species.     

Evaluation of optimum dietary protein-to-energy ratio in juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel)

An 8‐week feeding trial was conducted to estimate the optimum dietary protein to energy (P/E) ratio in juvenile olive flounder Paralichthys olivaceus. Eight experimental diets were formulated with two energy levels and four protein levels at each energy level. Two energy levels of 12.5 and 16.7 kJ g^?1 diets were included at crude protein (CP) levels of 25%, 30%, 35% and 45% with 12.5 kJ g^?1, and CP levels of 35%, 45%, 50% and 60% with 16.7 kJ g^?1. After 1 week of the conditioning period, fish initially averaging 8.1±0.08 g (mean±SD) were randomly distributed into the aquarium as groups of 15 fish. Each diet was fed on a dry‐matter basis to fish in three randomly selected aquariums at a rate of 3–5% of total wet body weight per day for 8 weeks. After 8 weeks of the feeding trial, weight gain (WG), feed efficiency ratio and specific growth rate of fish fed 45% CP with 16.7 kJ g^?1 energy diet were significantly higher than those from the other dietary treatments (P<0.05). WG of fish fed 12.5 kJ g^?1 energy diets increased with the increase of dietary protein levels. However, WG of fish fed 16.7 kJ g^?1 energy diets increased with the increase of dietary protein levels up to 45% CP and then decreased when fish fed 50% and 60% CP diets. Both dietary protein and energy affected protein retention efficiency and energy retention efficiency. Haemoglobin (Hb) of fish fed 35% and 45% CP diets with 12.5 kJ g^?1 energy were significantly high and not different from Hb of fish fed 45% and 50% CP diets with 16.7 kJ g^?1 energy. Haematocrit of fish fed 45% CP diet with 16.7 kJ g^?1 energy was significantly higher than those from fish fed 25% and 30% CP diets with 12.5 kJ g^?1 energy (P< 0.05). Based on the results of this experiment, we concluded that the optimum dietary P/E ratio was 27.5 mg protein kJ^?1 with diet containing 45% CP and 16.7 kJ g^?1 energy in juvenile olive flounder.     

Effects of starvation on kidney melano-macrophage centre in olive flounder, Paralichthys olivaceus (Temminck and Schlegel)

We investigated the influence of nutritional conditions on histological changes in melano‐macrophage (MM) accumulation in the kidney caused by long‐term starvation. These data were used to determine nutritional indices for olive flounder, Paralichthys olivaceus (Temminck and Schlegel). Experimental groups were established: control, fed and starved. All fish were fed daily with commercial feed at 1–3% of their total body weight for 2 weeks before the start of the experiment (initial control). The control group received feed at 1–3% of their total body weight and the fed group received feed ad libitum, while the starved group fasted throughout the experiment. Ten fish were removed from the initial control at the start of the experiment and each of the three groups every 4 weeks during the 12 weeks of the experiment to examine to investigate kidney MM accumulation. Kidney MMs with dark brown pigment were randomly distributed in the kidneys of starved fish, increasing rapidly after week 4, while deposition levels remained low throughout the experiment in the control and fed groups. These results suggest that catabolic tissue breakdown is a major factor contributing to the formation of pigments within MMs. These results suggest that the degree of MMs deposition in the kidney can be used as alternative indicators to identify starvation in wild and cultured olive flounder.     

Genetic relationship between broodstocks of olive flounder, Paralichthys olivaceus (Temminck and Schlegel) using microsatellite markers

For the first generation of a selective breeding programme, it is important to minimize the possibility of inbreeding. This mostly occurs by mating between closely related individuals, while proper mating can provide an opportunity to establish the base families with wide genetic variation from which selection for subsequent generations can be more effective. Genotyping with microsatellite‐based DNA markers can help us determine the genetic distances between the base populations. The genetic markers further facilitate the identification of the correct parents of the offspring (parentage assignments) reared together with many other families after hatching. We established a genetic analysis system with microsatellite DNA markers and analysed the genetic distances of three farmed stocks and a group of fish collected from wild populations using eight microsatellite markers. The averaged heterozygosity of the farming stocks was 0.826 and that of the wild population was 0.868. The hatchery strains had an average of 8.6 alleles per marker, which was less than a wild population that carried an average of 14.3 alleles per marker. Significant Hardy–Weinberg disequilibrium (HWDE) was observed in two farming stocks (P<0.05). Despite relatively low inbreeding coefficiency of the hatchery populations, the frequency of a few alleles was highly represented over others. It suggests that the hatchery stocks to some extent have experienced inbreeding or they originated from closely related individuals. We will develop a selective program using the DNA markers and will widen the usage of the DNA‐based genetic analysis system to other fish species.     

DNA polymorphism in the growth hormone gene and its association with weight in olive flounder Paralichthys olivaceus

ABSTRACT: Variation within the growth hormone gene and its association with growth trait in the olive flounder ( Paralichthys olivaceus ) was investigated. Based on Southern blot analysis and using various kinds of restriction endonucleases, it was demonstrated that the growth hormone gene exists as a single copy gene in the olive flounder. Polymorphisms of various lengths were also detected by Southern blot analysis, and by the subsequent digestion of the polymerase chain reaction (PCR) -amplified growth hormone gene fragment with Sau 3AI restriction enzyme. To study the possible association between variation in the growth hormone and weight, 60 progenies of the hatchery strain of three various sizes (large, medium and small weight) were selected and the entire genetic structure of the growth hormone gene was analysed. A total of 15 different genotypes was observed from the random association of six haplotypes. Significant heterogeneity of the growth hormone gene with haplotype and genotype frequencies was detected among the different-sized groups.