Dietary dehulled soybean meal as a replacement for fish meal in fingerling and growing olive flounder Paralichthys olivaceus (Temminck et Schlegel)

Two experiments were conducted to determine the optimum dietary inclusion level of dehulled soybean meal (DHSM) as a fish meal (FM) replacement in diets for olive flounder Paralichthys olivaceus. All the experimental diets were formulated to be isonitrogenous and isoenergetic to contain 50% crude protein (CP) and 16.7 kJ energy g^?1 diet. In the first experiment, eight diets were formulated to replace FM with DHSM at 0%, 10%, 20% without amino acid (AA) supplementation; 20%, 30%, 40% with AA supplementation and 30%, 40% with AA & attractant supplementation (DHSM₀, DHSM₁₀, DHSM₂₀, DHSM_20+AA, DHSM_30+AA, DHSM_30+AA+Att, DHSM_40+AA, DHSM_40+AA+Att respectively). Triplicate groups of 25 fish averaging 5.0±0.04 g (mean±SD) were fed one of eight experimental diets for 8 weeks. In the second experiment, six diets were formulated to replace FM with DHSM at 0%, 10%, 20%, 30% without attractant supplementation and 20%, 30% with attractant supplementation (DHSM₀, DHSM₁₀, DHSM₂₀, DHSM₃₀, DHSM_20+Att, DHSM_30+Att respectively). Triplicate groups of 15 fish averaging 45.5±0.08 g (mean±SD) were fed one of six experimental diets for 10 weeks. Based on growth performance, we concluded that DHSM could replace FM up to 20% without AAs (lysine and methionine) and attractant supplementation, and up to 30% with AAs and/or attractant supplementation in diets for fingerling and growing 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.     

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.     

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.     

Optimum feeding rates in juvenile olive flounder,Paralichthys olivaceus,at the optimum rearing temperature

Two feeding trials were carried out to determine the optimum feeding rates in juvenile olive flounder, Paralichthys olivaceus, at the optimum rearing temperature. Fish averaging 5.0 ± 0.11 g (mean ± SD) in experiment 1 and 20.2 ± 0.54 g (mean ± SD) in experiment 2 were fed a commercial diet at the feeding rates of 0%, 3.0%, 4.0%, 4.25%, 4.5% and 4.75% body weight (BW) day^?1 and satiation (5.52% BW day^?1) in experiment 1 and 0%, 1.0%, 2.0%, 3.0% and 3.5% BW day^?1 and satiation (4.12% BW day^?1) in experiment 2 at 20 ± 1 °C. Both feeding trials lasted for 2 weeks. Results from experiment 1 indicated that weight gain (WG) and specific growth rate (SGR) of fish fed to satiation were significantly higher than those of fish fed at other feeding rates while feed efficiency (FE) and protein efficiency ratio (PER) of fish fed at 4.25% BW day^?1 were significantly higher than those of fish fed to satiation and fish fed at 3.0% BW day^?1 (P < 0.05). In experiment 2 WG, SGR and PER leveled out after the feeding rate of 3.5% BW day^?1 whereas FE reached a plateau at 3.0% BW day^?1. anova of FE indicated that the optimum feeding rates in 5.0 and 20 g juvenile olive flounder could be 4.25% and 3.0% BW day^?1, respectively. Broken line analysis of WG suggested the optimum feeding rates of 5.17% and 3.47% BW day^?1 in 5.0 and 20 g fish, respectively. Therefore, these results indicated that the optimum feeding rates could be >4.25 but <5.17% BW day^?1 for 5.0 g, and it could be >3.0 but <3.47% BW day^?1 for 20 g size of juvenile olive flounder at the optimum rearing temperature.     

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.     

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.     

Pathogenicity of Edwardsiella tarda to olive flounder, Paralichthys olivaceus (Temminck & Schlegel)

The LD50 and cytotoxic and enzymatic activities of both cells and extracellular products (ECPs) of eight Edwardsiella tarda strains were determined and their bacterial superoxide dismutase gene (sodB) and catalase gene (katB) were sequenced. Strains were also examined for their ability to resist the immune responses of olive flounder, Paralichthys olivaceus. LD50 values of strains (FSW910410, KE1, 2, 3, 4, 5 and 6) in olive flounder ranged between 10(2.5) and 10(5.3) cfu (colony forming units) per fish. Unlike the avirulent strain SU100 (LD50>or=10(7)), all pathogenic strains were able to survive in flounder serum and head kidney leucocytes (except for KE2). The virulent strains possessed type I sodB and katB, whereas SU100 had type II sodB but not katB. However, there was no difference between avirulent and virulent strains in haemolytic and cytotoxic activities. The results of this study demonstrated that the ability of E. tarda to resist complement activity and phagocytosis is conferred by its superoxide dismutase and catalase, which thus play an essential role in the pathogenicity of this bacterium. In addition genotyping of sodB and kat B proved to be a very useful tool to distinguish virulent from avirulent strains.     

牙鲆微卫星标记的开发及多态性分析

将牙鲆基因组DNA经限制性内切酶RsaⅠ和BstUI双酶切后采用FIASCO法构建酶切片段基因组文库。共挑取269个克隆,177个为阳性克隆,阳性克隆率为65.79%。经测序后获得191条微卫星序列,其中完美型占74.35%;非完美型占14.66%;混合型占10.99%。用引物设计软件Primer Premier 5.0设计引物153对,挑选其中的50对合成并在32个野生牙鲆个体中进行扩增,共31个位点具有多态性,统计结果后使用POPGENE软件进行分析,平均等位基因个数为3.939 4,平均有效等位基因数为3.052 2,平均观测杂合度为0.650 1,平均期望杂合度为0.586 6,各引物的Hardy-Weinberg平衡指数在0.012 587~0.984 917变动。这些筛选出的多态性微卫星标记可应用于进一步的牙鲆遗传多样性分析、家系分析及遗传图谱的构建等工作中。     

Effects of dietary fish oil replacement by microalgae raw materials on growth performance,body composition and fatty acid profile of juvenile olive flounder,Paralichthys olivaceus

Replacing dietary fish oil with DHA‐rich microalgae Schizochytrium sp. and EPA‐rich microalgae Nannochloropsis sp. for olive flounder (Paralichthys olivaceus) was examined. Three experimental isonitrogenous and isolipidic diets with lipid source provided by 50% fish oil (F50S50), 50% (M50F25S25) and 100% microalgae raw material (M100) respectively were compared with a soybean oil (S100) diet as control. Triplicate groups of olive flounder juveniles (16.5 ± 0.91 g) were fed the experimental diets, and a group was fed the control diets for 8 weeks in a recirculation system. Results showed feed efficiency and growth performance were not significantly changed when fish oil (FO) was totally substituted by soybean oil (SO) or microalgae raw material (MRM). The whole‐body composition, lipid content of liver and muscle, and lipid composition of plasma were not significantly influenced by the total substitution of FO by MRM. The polyunsaturated fatty acids (PUFA) content of muscle and liver declined in fish fed S100 diet, whereas it was not significantly reduced in fish fed M50F25S25 and M100 diets. The total substitution of FO by MRM not only maintained the levels of arachidonic acid, EPA or DHA but also increased n‐3/n‐6 ratio. In conclusion, MRM as the sole lipid source is sufficient to obtain good feed efficiency, growth performance and human health value in olive flounder juveniles.     

牙鲆微卫星标记的筛选及群体多态性分析

以牙鲆（Paralichthys olivaceus Temminck et Schlegel）为材料，基因组DNA经Sau3A Ⅰ进行部分酶切后，选取400～1200bp大小的片段连接到经BamH Ⅰ酶切的pUC19质粒中，连接产物转化大肠杆菌DH5a，构建牙鲆酶切片段基因组文库。采用菌落杂交的方法，以（AC）10、（AG）10为探针从文库中筛选阳性克隆16个，共得到20个微卫星序列，其中完全型13个，不完全型5个，复合型2个。对其中18个进行引物设计，有11对引物扩增出目的片段，其中8对引物在群体中具有扩增多态性。在威海野生牙鲆30个个体中，各座位上得到的等位基因数为4～19个，观测杂合度（Ho）为0．4138～0．9655。其中有3对引物扩增的等位基因数超过17个，表现出高度多态性。本研究旨为进一步的牙鲆遗传多样性分析、家系分析及遗传图谱的构建等提供基础依据。     

Detection of antigenic proteins expressed by lymphocystis virus as vaccine candidates in olive flounder, Paralichthys olivaceus (Temminck & Schlegel)

Although the major capsid proteins (MCPs) of lymphocystis disease virus (LCDV) have been characterized, little is known about the host-derived immune response to MCPs and other LCDV antigenic proteins. To identify antigenic proteins of LCDV that could be used as vaccine candidates in olive flounder, Paralichthys olivaceus, we analysed the viral proteins responsible for its virulence by applying immuno-proteomics. LCDV proteins were separated by one-dimensional gel electrophoresis, transferred to polyvinylidene difluoride membrane, and probed with homogeneous P. olivaceus antisera elicited by LCDV natural infection and vaccination with formalin-killed LCDV. Four immune-reactive proteins were obtained at 68-, 51-, 41- and 21 kDa using antisera collected from natural infection while two proteins at 51- and 21 kDa exhibited response to antisera from vaccinated fish, indicating that the latter two proteins have vaccine potential. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nanoelectrospray MS/MS, the 51 and 21 kDa proteins were identified as MCP and an unknown protein, respectively.     

Effect of phenanthrene on haematological parameters in olive flounder, Paralichthys olivaceus (Temminch et Schlegel)

Effects of phenanthrene on haematological parameters in olive flounder, Paralichthys olivaceus, were studied by exposing fish to water‐borne phenanthrene (0.5, 1 and 2 μM) for 4 weeks. Red blood cell count, haemoglobin concentration, haematocrit levels decreased with increasing exposure time of phenanthrene to the fish. Plasma potassium level increased significantly in the experimental group but the levels of sodium and chloride were significantly decreased compared with control. There was no significant change in plasma magnesium, calcium and phosphate in fish of all treated groups compared with the control. The most profound physiological disturbance in phenanthrene‐exposed fish was a decrease in blood osmolality. Malondialdehyde level in the plasma in the phenanthrene group (2 μM) was 1.4‐fold higher following 4‐week exposure compared with the control group. Acetylcholinesterase activity decreased as the concentration of phenanthrene increased, reaching 65.9% of the control value at the highest dose at the 4th week. A key finding from this study is that olive flounder exposed to phenanthrene at concentrations more than 1.0 μM are likely to experience negative impacts on haematological parameters.     

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.     

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.     

Evaluation of shrimp or tilapia protein hydrolysate at graded dosages in low fish meal diet for olive flounder (Paralichthys olivaceus)

A study was carried out to examine and optimize the inclusion levels of shrimp hydrolysate (SH) or tilapia hydrolysate (TH) in low fishmeal diets for olive flounder (Paralichthys olivaceus). A fishmeal (FM)‐based diet was considered as a high FM (HFM) diet, and a diet containing soy protein concentrate (SPC) as a FM replacer at 50% substitution level was regarded as a low FM (LFM) diet. Six other experimental diets were prepared by dietary supplementation of SH or TH to LFM diet at different inclusion levels of 15 g/kg, 30 or 45 g/kg in the expense of FM (designated as SH‐1.5, SH‐3.0, SH‐4.5, TH‐1.5, TH‐3.0 and TH‐4.5, respectively). After 10 weeks of a feeding trial, growth performance and feed utilization efficiency of fish were significantly higher in fish fed HFM, SH‐3.0, SH‐4.5, TH‐1.5 and TH‐3.0 diets compared to those of fish fed LFM diet. Intestine diameter, villus height and goblet cell counts of fish were significantly increased by dietary inclusion of SH or TH into LFM diet. Dry matter and protein digestibility of diets were significantly improved by SH or TH incorporation. Innate immunity of fish was significantly enhanced by dietary SH or TH supplementation into LFM diet. Disease resistance of fish was significantly increased against Edwardsiella tarda by dietary inclusion of SH and TH at the highest inclusion level (45 g/kg). The optimum inclusion level of SH or TH in a SPC‐based LFM diet could be ~30 g/kg and 15–30 g/kg, for olive flounder.     

Lymphocystis disease virus persists in the epidermal tissues of olive flounder, Paralichthys olivaceus (Temminch & Schlegel), at low temperatures

Olive flounder artificially infected with lymphocystis disease virus (LCDV) were reared at 10, 20 and 30 °C for 60 days, to compare LCD-incidence. In the fish reared at 20 °C, lymphocystis cells appeared on the skin and fins at 35 days post-challenge, and the cumulative LCD-incidence was 80% at 60 days. High levels of LCDV, with a mean polymerase chain reaction (PCR) titre of 10⁶ PCR-U mg⁻¹ tissue, were detected in the fins and skin of LCD-affected fish at 20 °C, but were not detected in the spleen, kidney, brain and intestinal tissues of these fish. No LCD clinical signs were observed in the fish reared at 10 °C and 30 °C; however, a low level of LCDV (10³ PCR-U mg⁻¹ tissue) was detected in the fins and skin of these fish. By increasing the rearing temperature from 10 to 20 °C, lymphocystis clusters appeared on the skin and fins of the fish with no previous LCD clinical signs within 33 days after the temperature change. It was shown that permissive cells for LCDV infection exist in the epidermis of olive flounder. At low temperatures, small amounts of LCDV were able to persist over a period extended for a further 45 days in the fish epidermis, even though the fish showed no LCD clinical signs. The optimum growth temperature of LCDV is near 20 °C.     

Influences of dietary distillers dried grain level on growth performance,body composition and biochemical parameters of juvenile olive flounder (Paralichthys olivaceus)

This study was conducted to investigate the influence of dietary distillers dried grain (DDG) level on growth and body composition of juvenile olive flounder (Paralichthys olivaceus). Five diets (designated as DDG0, DDG7, DDG14, DDG21 and DDG28) were prepared to contain 0, 70, 140, 210 and 280 g kg^?1 DDG. Three replicate groups of fish averaging 11.6 ± 0.1 g were fed one of the diets for 8 weeks. Weight gain of fish fed diets containing 70–280 g kg^?1 DDG was not different to that of fish fed DDG0 diet (P > 0.05). Feed efficiency of fish fed DDG21 and DDG28 diets was lower than that of fish fed DDG0 diet (P < 0.05). The antioxidant enzyme and digestive enzyme activities were not affected by dietary DDG levels. The results of this experiment suggested that DDG is a valuable ingredient as a candidate to replace wheat flour in the diet and could be used up to 280 g kg^?1 for the growth of olive flounder. On the other hand, if lower feed efficiency of fish fed 210–280 g kg^?1 DDG is considered, up to 140 g kg^?1 DDG in diet is recommended for optimum feed utilization of fish.     

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.