Protein Requirements of the Freshwater Prawn Macrobrachium rosenbergii Evaluated by the Factorial Method

The dietary protein requirement of the freshwater prawn Macrobrachium rosenbergii to obtain the maximum body protein retention was assessed by the estimated protein needed daily for maintenance (M) and body protein increment (G). The body protein increment was estimated by quantifying body nitrogen increments when the prawn was fed high‐protein diets composed primarily of casein. The protein required for maintenance was estimated by regressing weight gains in feeding experiments using graded dietary levels of casein. The true daily retention (R) or increase in body protein of the prawn corresponds to the sum of M and G. The values of M, G, and R determined in the present study were 3.86, 0.24, and 4.10 g/kg body weight/d, respectively. On the basis of the daily body protein increment and the net protein utilization of casein (58%), dietary protein requirement of the juvenile M. rosenbergii for maximum body protein retention was suggested to be about 7.1 g/kg body weight/d.     

Studies on the Nutrition of Spotted Sand Bass Paralabrax maculatofasciatus: Effect of the Dietary Protein Level on Growth and Protein Utilization in Juveniles Fed Semipurified Diets

Two feeding trials were conducted to determine the digestibility of a casein-based semi-purified diet and the effects of different protein levels on growth and protein use of spotted sand bass Paralabrax maculatofasciatus juveniles. For trial I, a semipurified diet with vitamin-free casein as the sole source of protein was fed three times a day to apparent satiation, for a period of 20 d. Feces were collected by siphoning each tank. The digestibility of the experimental diet was high: 97% for protein, 89% for lipids, and 84% for gross energy, whereas that of organic matter was 78%. For trial II, seven diets were formulated using vitamin-free casein at graded levels (25, 30, 35, 40, 45, 50, and 55% protein). Triplicate tanks for each dietary treatment were stocked with fish and fed by hand three times a day to apparent satiation for 6 wk. Perfomance of fish fed the different diets was evaluated for survival, percent weight gain, specific growth rate, feed conversion ratio, and protein efficiency ratio. Survival was 100% for all treatments. Growth of spotted sand bass juveniles increased as the dietary protein increased, but no evidence of reaching a plateau was found. The daily feed intake values showed an inverse relation to the protein content of the diets. The feed conversion ratio did not differ among diets containing 40% protein or greater. The results indicate that spotted sand bass juveniles with 2.5-g mean weight need at least 55% dietary protein for best growth when casein is the sole protein source. However, in terms of feed conversion ratio, the requirement apparently could be lower.     

鱼类病毒性出血性败血症病毒基质蛋白的原核表达及其亚细胞定位

为了对鱼类病毒性出血性败血症病毒(viral hemorrhagic septicemia virus, VHSV)基质蛋白(matrix protein, M)进行功能研究,本实验通过PCR扩增了M基因全长序列,将其克隆至原核表达载体pET-32a(+),转化至大肠杆菌Rosetta(DE3)感受态细胞后进行IPTG诱导表达,将纯化后的重组蛋白免疫BALB/c小鼠制备多克隆抗体,采用间接ELISA检测抗体效价,并运用Westernblot和间接免疫荧光检测抗体特异性。结果显示,M基因全长为606bp,IPTG诱导得到的融合蛋白主要以包涵体的形式存在,大小约为36 kD,比预计略小。间接ELISA检测抗体效价大于1:102400,Westernblot检测显示该抗体可以特异性识别纯化的融合蛋白和VHSV感染的鲤上皮瘤(epithelioma papulosum cyprini, EPC)细胞中的M蛋白。间接免疫荧光结果显示M蛋白多抗能识别感染VHSV的EPC细胞中的M蛋白,且M蛋白主要定位于细胞质和细胞膜。本研究中M蛋白多克隆抗体的制备将有助于开展M蛋白的功能研究及疾病的免疫学诊断。     

Studies on the effect of increasing proportions of sucrose or gelatinized maize starch in diets for rainbow trout (Salmo gairdneri, R.) on the utilization of dietary energy and protein

Trial 1: eight diets containing between 25 and 54% sucrose and correspondingly between 47 and 23% crude protein were fed to replicate groups of trout for 44 or 45 days. Feed conversion increased only when the sucrose content was above 36% and crude protein correspondingly below 40%. Decreasing the protein content by increasing the proportion of sucrose improved the protein efficiency ratio (PER) between 47 and 40% protein; PER values lower than for the diet containing 47% crude protein were obtained only when the protein concentration was less than 36%.

Trial 2: protein of a fish meal/casein diet was replaced by 10–40% sucrose or gelatinized maize starch. All diets were fed to replicate groups of trout for 60 days. Although gelatinized maize starch did not affect the conversion ratio, with increasing proportions of sucrose in-creasing amounts of feed were required per unit of weight gain. PER values increased with increasing proportions of either carbohydrate. No clear differences could be found in body composition. Efficiency of utilization of gross energy was between 40 and 43% for the basal diet and diets containing gelatinized maize starch, whereas it was reduced to about 31% by increasing the proportion of sucrose to 40%. The percentage of dietary N retained rose from 23 to 29% by increasing sucrose, and to 44% by increasing gelatinized maize starch in the diet.     

鲤肠道小肽转运载体PepT1多克隆抗体的制备及其组织表达分析

为从蛋白水平研究小肽转运载体(PepT1)在鲤(Cyprinus carpio L.)不同组织中的表达及分布规律,本研究采用PCR法获得PepT1 cDNA片段,并转化至大肠杆菌Rosetta,对目标多肽进行原核表达,将Pep T1重组蛋白纯化后免疫新西兰长耳兔(Oryctolagus cuniculus),获取兔抗鲤PepT1多克隆抗体。采用ELISA检测抗体效价,免疫组化检测PepT1的组织表达情况,并用荧光实时定量PCR技术检测PepT1转录水平组织表达情况。结果显示,目标多肽分子量约为28 kD;抗体效价达到4×10~5。PepT1在鲤的前肠、中肠、后肠、脾、肝胰脏和肾中均有表达。肠道组织PepT1的高表达与其主要完成食物中肽的吸收功能密切相关,且吸收部位主要集中在前肠和中肠;肾中PepT1免疫染色阳性区域也较为明显,这与肾小管基底膜存在对短肽的重吸收功能相关。此外,肝胰脏和脾PepT1也有一定量的表达,可能与这两个重要器官代谢旺盛有关。本研究制备的兔抗鲤PepT1多克隆抗体能够有效识别鲤各组织中的PepT1蛋白,在后续研究中亦可用于其他鱼类PepT1转运蛋白的表达定位和定量研究。     

Characterization of the monoclonal antibody specific to the ORF72 protein of koi herpesvirus and cellular distribution analysis of the viral protein

Koi herpesvirus (KHV) is an emerging pathogen of koi and common carp that causes a severe disease and mass mortality of infected fish. The KHV ORF72 protein is an important capsid protein that has been suggested to be a candidate for the development of diagnostic reagents and KHV vaccines. The purpose of this study was to clone and express the KHV ORF72 gene for further preparation of a specific monoclonal antibody (mAb) and to analyse cellular distribution of the viral protein. The mAb 3E1 could specifically recognize the expressed ORF72 protein of transfected cells by indirect immunofluorescence, and the antigenic site recognized by the mAb 3E1 was mapped to the region of N-terminal 124 residues of KHV ORF72. This mAb was further demonstrated to specifically detect the KHV-infected fish tissue by immunohistochemistry, thereby suggesting its high diagnostic potential. In addition, the cellular distribution analysis of the KHV ORF72 protein revealed that the region of amino acid residues 125–247 was related to mitochondrial localization and proliferation. Furthermore, a putative nuclear export signal (NES) of ORF72 at the residues 201–212 was confirmed on the basis of its function associated with NES activity.     

添加小麦蛋白对马面鲀鱼糜凝胶强度影响的研究

<正> 自从60年代初日本的西谷等人开发冷冻鱼糜制造技术以来,鱼糜制品在日本得到了很大的发展。目前日本冷冻鱼糜年产量100万吨左右,鱼肉香肠、鱼肉火腿年产量10万吨左右。近年来,鱼糜制品技术日益引起世界各国的重视,美国、欧洲和亚洲的泰国、南朝鲜,我国台湾等都在研究开发鱼糜制品。鱼肉火腿,鱼肉香肠作为鱼糜制品中的     

传染性造血组织坏死病毒核衣壳蛋白的原核表达及抗原性分析

应用RT-PCR方法扩增了长度为1176 bp的IHNV-ZYX株编码核衣壳(N)蛋白基因,将N基因克隆至原核表达载体pET30b,并在大肠杆菌Rosetta(DE3)中得到了表达.通过SDS-PAGE分析表明,重组菌诱导后得到了预期大小约48 KD的N蛋白,与理论值相符;提取N蛋白的包涵体,并制备抗血清.间接ELIS...     

A Standard Reference Diet for Crustacean Nutrition Research. II. Selection of a Purification Procedure for Production of the Rock Crab Cancer irroratus Protein Ingredient

Obtaining reliable results from comparative crustacean nutrition studies requires a dietary reference protein which is available world-wide and is nutritionally optimal for growth and survival. Traditional protein sources include "vitamin free" casein, egg albumin, whole egg protein, soybean protein, and others. These have proven to be inadequate for juvenile lobster ( Homrus americanus ). Purified rock crab ( Cancer irroratus ) protein is proposed to be nutritionally superior to casein for inclusion in semi-purified lobster diets.
Eighteen week feeding trials with juvenile H. americanus were conducted to compare diets containing protein obtained from rock crabs ( Cancer irroratus ), and produced by several different purification processes with a control diet having casein as its primary protein source. Diets containing purified rock crab protein were found to be superior to the control diet. Among the differently derived purified rock crab protein products, the precooked, isopropanol-purified was the most effective dietary ingredient.     

Oxygen Consumption and Ammonia Excretion of the American Eel Anguilla rostrata Fed Diets with Varying Protein Energy Ratios and Protein Levels

The effect of protein/energy ratio and protein level on utilization of protein and energy as indicated by oxygen consumption and ammonia production was investigated. When protein/energy ratio of the diet was optimum, there was no significant increase in ammonia production, indicating no increase in catabolism of protein for energy as protein levels increased. However, when the protein/ energy level was below optimum, there was a significant ( P < 0.05) increase in ammonia production as protein increased, indicating increased catabolism of protein for energy. However, there was no significant increase in oxygen consumption rate.     

中华绒螯蟹精巢特异表达蛋白DMRT-like:抗体制备及免疫鉴定

Dmrt是性别调控因子Doublesex和Mab-3的相关基因,近年报道了中华绒螯蟹EsDmrt-like只在精巢中表达,为了验证EsDMRT-like蛋白是否在中华绒螯蟹精巢中特异表达及其功能,根据中华绒螯蟹EsDmrt-like基因序列,构建重组质粒pET-32a-EsDmrt-like,转化大肠杆菌BL21,经融合表达和SDS-PAGE分析表明,融合蛋白主要以包涵体形式存在,分子量约为46ku.利用Ni柱亲和纯化融合蛋白免疫家兔,制备获得EsDMRT-like多克隆抗体.Western-blotting检测表明该抗体既能特异地识别重组蛋白,又能特异识别精巢中EsDMRT-like蛋白,并且该抗体仅在精巢中检测到EsDMRT-like蛋白的表达,分子量约为52 ku,为预期单体分子量的二倍.Western-blotting检测变性后精巢总蛋白,该抗体能识别52和34 ku两条条带,证明了二聚体的存在.这一结果暗示EsDMRT-like可能通过形成二聚体形式调控中华绒螯蟹精巢发育.     

Effect of the use of the microalga Spirulina maxima as fish meal replacement in diets for tilapia, Oreochromis mossambicus (Peters), fry

The present study addresses the use of the microalga Spirulina maxima as a protein source in diets for tilapia, Oreochromis mossambicus (Peters), fry. Animal protein was replaced with algae protein at ratios of 20%, 40%, 60%, 80% and 100%, and the substitution effect was compared with a control diet in which fish meal was the sole protein. An additional 100% spirulina protein diet was supplemented with phosphorous to test for possible mineral deficiency in the plant-protein-based diet. The six treatments were tested in triplicate in a closed-recirculating system where the fish were fed by hand at 6% of their body weight. After a 9-week feeding period, the growth rate and protein utilization of fish fed the diet with 20% and 40%Spirulina were elevated and not significantly different (P>0.05) from those fed the control diet. Further increases in the alga protein content significantly decreased the growth and feeding performance. The addition of P to the 100%Spirulina diet slightly improved performance in comparison to the same diet without P. None of the treatments produced any clear adverse effects on carcass composition. It is observed that Spirulina can replace up to 40% of the fish meal protein in tilapia diets.     

Fast growth, protein turnover and costs of protein metabolism in yolk-sac larvae of the African catfish (Clarias gariepinus)

In fish larvae the costs of rapid growth may be accommodated by a decrease in the rate of protein turnover or by a reduction in the costs of protein synthesis. Protein growth, synthesis and degradation were measured in yolk-sac larvae of Clarias gariepinus and the costs of protein synthesis and protein growth were estimated. Growth rates were over 100% protein weight day^-1. Protein synthesis retention efficiency (retained protein per unit of synthesis) was estimated to be 69.6%, a value comparable to that of larger fish. The larvae used 43% of their oxygen consumption for protein synthesis. Nevertheless, protein synthesis costs were close to theoretical minima. Therefore, the high growth rates of catfish yolk-sac larvae seem to be possible through minimisation of the costs of protein synthesis. These low costs are associated with high rates of protein synthesis (138%protein weight day^-1), and elevated RNA concentrations (107 µg RNA mg^-1protein), which together suggest very high RNA efficiencies (12.9 g protein synthesized g^-1RNA day^-1).     

Effect of extrusion processing on the nutritive value of canola meal for chinook salmon (Oncorhynchus tshawytscha) in seawater

This study was conducted to evaluate three canola protein products as partial replacements of steam-dried whole herring meal (HM) in a practical diet for juvenile chinook salmon in seawater. Groups of 30–40 g chinook salmon held in 9.5–12°C seawater on a natural photoperiod were fed one of seven equivalent protein (390 g kg⁻¹) and isoenergetic (20 MJ gross energy kg⁻¹) diets to satiation. Each of three test protein sources, namely, commercial canola meal (CM), low-temperature extruded CM (90°C; LT), and high-temperature extruded CM (150°C; HT) comprised about 120 and 240 g kg⁻¹ of dietary protein by replacement of 15 and 30%, respectively, of the HM protein in the control diet. Chinook salmon growth and feed intake were markedly depressed by replacing HM protein with a high level of CM, but these responses were not lowered by replacement with LT and HT. The phytic acid content in commercial canola meal was reduced by about 10 and 30% from the original level by extrusion cooking at low (90°C) and high (150°C) temperature, respectively. Therefore, the diets containing HT still had significant amounts of phytic acid. Terminal whole-body zinc contents were inversely related to the dietary levels of the different canola protein products. Thyroid function was altered by CM but this effect was offset by heat treatment. The results suggest that extrusion cooking of CM improves its nutritive value for chinook salmon in seawater. Indeed, it was found that HT could comprise 240 g kg⁻¹ of the dietary protein without adversely affecting performance.     

三疣梭子蟹幼蟹配合饲料中蛋白质、脂肪、粗纤维适宜含量研究

以初始平均体重为7.1 g的三疣梭子蟹(Portunus trituberculatus)幼蟹为试验对象,以粗蛋白、粗脂肪和粗纤维为试验因素,各设3个水平,采用正交试验设计L9(34),配制成9种饲料,在室内水族箱进行为期4周的饲养试验,以探讨三疣梭子蟹幼蟹配合饲料中蛋白质、脂肪和粗纤维的适宜含量.结果表明,三疣梭子蟹幼蟹配合饲料中蛋白质、脂肪和粗纤维的适宜含量分别为40%、6%～8%和≤5%时,三疣梭子蟹幼蟹增重率和蜕壳率高,饲料系数低和蛋白质效率较高.     

Increase in the plant protein ratio in the diet of white shrimp,Litopenaeus vannamei (Boone), using Bacillus subtilis E20‐fermented soybean meal as a replacement

This study aimed to improve the nutritional value and utilization of soybean meal (SBM) used as feed ingredient for shrimp by an approach of solid‐state fermentation with Bacillus subtilis E20. The protein content and B. subtilis E20 proliferation significantly increased, as the initial moisture increased from 30% to 50% during fermentation. Compared with SBM, the protein content of FSBM increased by 19% after fermentation, accompanied by an increase of 18.75% in the total hydrolyzed amino acids. The free amino acid profile and content in FSBM also obviously increased by 374.9% compared with SBM. The FSBM is a good substitute for fish meal (FM) in a diet with 37% protein and 7% lipid content. The maximal replacement levels of FM in shrimp diet with SBM and FSBM were 37.42% and 61.67%, respectively, based on the feed efficiency estimated by a broken‐line analysis. The B. subtilis E20‐FSBM can be a potential protein source used as a replacement for FM in shrimp diet.     

Effect of dietary lipid level on protein utilization and the size and proximate composition of body compartments of Colossoma macropomum (Cuvier)

Juvenile Colossoma macropomum were fed ad libitum diets containing either 30% or 40% protein, while the dietary lipid level varied between 5% and 20%. Growth and protein utilization efficiency increased with the dietary lipid level. However, the economic feasibility of the addition of extra dietary lipid to C. macropomum diets is questionable, as for every extra gram of protein deposition it was necessary to replace 20–25 g of dietary carbohydrates by lipids. Moreover, increased dietary lipid levels resulted in increased lipid deposition. In C. macropomum, feed uptake was regulated by the dietary protein level and unaffected by the dietary lipid level. Dissection of the body into head, viscera and trunk revealed that 45–48% of the body lipids were stored in the trunk, independent of diet composition. A positive relation between dietary and body protein level was confirmed in this experiment. However, the dissection of the body revealed that the body protein concentrations in head + viscera and trunk are not changed significantly by the dietary protein level. The higher protein contents offish fed higher-protein diets are due to an increase in the relative weight of the trunk (muscle) to the total body weight.     

Evaluation of an in vitro digestibility technique for the prediction of protein digestibility in the South African abalone, Haliotis midae L.

A pH-stat multienzyme in vitro digestibility assay was investigated for its efficacy in predicting apparent protein digestibility coefficients in abalone diets. Linear regression analysis between in vitro digestibility estimates and in vivo digestibility coefficients obtained from abalone, revealed that the technique could be used to reliably predict apparent protein digestibility. Maximal predictability of the system was obtained when protein sources were analysed according to origin – animal or plant. The technique was used to assess the apparent protein digestibility of 34 potential protein sources for use in formulated feeds for Haliotis midae .     

Development of plant‐based diets and the evaluation of dietary attractants for juvenile Florida pompano,Trachinotus carolinus L.

Sustainable and profitable commercial aquaculture production of marine fish species is dependent on the development of sustainable protein sources as substitutes for expensive animal meals such as fishmeal (FM). Previous Florida pompano Trachinotus carolinus studies have indicated that poultry by‐product meal (PBM) and meat and bone meal can be used to produce a FM‐free diet if suitable levels of nutrients (such as taurine) are included in the diets. In this study, we attempted to develop an all‐plant protein diet by removing the animal proteins in practical diets for pompano by substituting back select ingredients. A series of eight FM‐free diets were formulated, four systematically replaced soybean meal (SBM) with soy protein concentrate (SPC) and four replaced PBM with SPC. Based on the results, there is no clear disadvantage to the use of SPC as a replacement for SBM. However, the complete removal of PBM resulted in reduced performance. Two additional growth trials were conducted to supplement additional amino acids including glycine, valine and histidine, a proprietary chemical attractant mix, fish protein concentrate and squid hydrolysate to improve the growth of pompano when fed all‐plant protein diets. The only improvement in performance occurred with the squid hydrolysate. These results demonstrate that using soybean meal, soy protein concentrate and corn gluten meal as the primary protein sources, a plant‐based feed formulation can be developed, but the removal of all animal proteins is not yet feasible.     

The effects of partial replacement of fishmeal protein by hydrolysed feather meal protein in the diet with high inclusion of plant protein on growth performance,fillet quality and physiological parameters of Pengze crucian carp (Carassius auratus var. Pengze)

Goal of this research is to explore the effects of graded dietary levels of hydrolysed feather meal (HFM) protein as an alternative of fishmeal protein on Pengze crucian carp (Carassius auratus var. Pengze). Five isonitrogenous (35% crude protein) diets were formulated as Control diet and four other diets F15, F30, F45 and F60 wherein 15%, 30%, 45% and 60% fishmeal protein replaced by HFM protein fed for 70 days. The results showed that fish growth was unaffected up to 45% fishmeal protein replacement. Dietary HFM protein could reduce the crude protein composition of body and further affect fillet quality through a significant increase in springiness, gumminess, chewiness and/or resilience. Activity of digestive enzymes (amylase, lipase and tryptase) increased as the fishmeal protein replacement increased, whereas a higher replacement level had negative impacts on absorptive capacity of intestine by decreasing its absorptive area, following by a reduction in concentration of lactate dehydrogenase, total cholesterol, high‐density lipoprotein, low‐density lipoprotein and ammonia in serum. Although intestinal antioxidant parameters were elevated by dietary HFM protein , the content of malondialdehyde in intestine was also positively associated with the protein replacement level. Overall, our results indicate that up to 45% of fishmeal protein could be replaced by HFM protein in diet without significant impact on the growth of Pengze crucian carp, whereas the crude protein content of whole body, fillet quality and oxidative status would significantly be affected when the replacement level of fishmeal protein reaches 30% together with a negative effect on the intestinal histology.