竹叶抗氧化物浸泡-乳清分离蛋白涂膜对冷藏富硒虹鳟蛋白质理化性质的影响

为从蛋白质水平解释竹叶抗氧化物浸泡-乳清分离蛋白涂膜对富硒虹鳟的保鲜机理,实验将鲜活富硒虹鳟去除头、内脏和表皮,切块后浸泡在1.0%的竹叶抗氧化物（AOB）溶液中（去离子水作为对照）,再进行乳清分离蛋白（WPI）溶液涂膜,25℃下鼓风吹干表面水分后置于4℃冰箱中贮藏1、4、7、10和13 d。测定鱼肉中肌原纤维蛋白和TCA-可溶性肽的含量、蛋白质的分子内价键,以及肌原纤维蛋白的总巯基含量、表面疏水性和二级结构,分析涂膜对冷藏富硒虹鳟蛋白质理化性质的影响。结果显示,冷藏期间,WPI组的肌原纤维蛋白含量和二级结构中β-折叠的含量显著高于对照组。冷藏1～7 d,WPI组的TCA-可溶性肽和二硫键含量以及表面疏水性指数（H₀）显著低于对照组,总巯基含量呈相反趋势,但随着冷藏时间的延长这些差异逐渐减弱。冷藏13 d时,WPI组与对照组无显著差异。3个实验组中,1.0%AOB+WPI组的肌原纤维蛋白、总巯基、氢键、二级结构中α-螺旋和β-折叠的含量在冷藏期间最高,H₀、TCA-可溶性肽、二硫键和二级结构中β-转角含量最低。WPI在鱼块表面形成一层薄膜,可...     

草鱼IFNa和IFNd重组蛋白表达、纯化及单克隆抗体制备

为系统研究草鱼I型干扰素的合成、分泌和免疫功能,本研究在大肠杆菌中表达并提纯了草鱼IFNa（CiIFNa）和IFNd（CiIFNd）重组蛋白。将CiIFNa和CiIFNd成熟肽分别克隆到pET-21d或pEHISTEVb表达载体上,并转化到大肠杆菌中;IPTG诱导表达得到CiIFNa和CiIFNd成熟肽的包涵体,经过盐酸胍变性、蛋白复性和浓缩后,利用AKTA分子筛层析获得了纯度较高的重组蛋白。用重组蛋白免疫小鼠,通过PEG法诱导得到杂交瘤细胞;将稳定分泌抗体的阳性细胞株的细胞悬液注射入小鼠腹腔,制备腹水抗体并进行纯化。本研究纯化了草鱼CiIFNa和CiIFNd各2株抗体,并采用SDS-PAGE、ELISA、Western blot和免疫荧光法对其进行了较全面的鉴定。研究结果表明CiIFNa和CiIFNd单克隆抗体特异性好、效价高,能够特异识别在大肠杆菌和真核细胞中表达的重组蛋白,且不存在CiIFNa和CiIFNd分子间的交叉识别。本研究制备的单克隆抗体为深入研究草鱼干扰素的细胞来源和蛋白表达规律奠定基础。     

灌喂必需氨基酸模式溶液对草鱼全鱼_省略_肌肉_肝胰脏蛋白质合成代谢的影响

采用大剂量食道灌喂DL－[4-^3H]-Phe，研究不同EAA模式下，20－30g体重草鱼种的全鱼和肌肉，肝胰脏的蛋白质合成代谢，结果表明：（1）分别缺乏Lys,Met,Trp Arg的4种极端不平衡EAA模式，同草鱼肌肉EAA模式即相对平衡的基础模式相比，肌肉，肝胰脏及全鱼的蛋白质合成速率（FSR）均显著降低，其降低程度高低依次为Met,Lys,Arg,Trp;(2)肝胰脏，肌肉人的蛋白质合成代谢与整体蛋白质合成代谢对EAA模式的改变产生相同的变化趋势，并且三者蛋白质合成代谢速率为肝胰脏＞全鱼＞肌肉；（3）EAA模式的Lys,Met,Trp,Arg4因素中，对肌肉，肝胰脏蛋白质合成速率影响最大的是Met,其余依次为Lys,Arg,Trp。     

Inhibition of protease activities in discus Symphysodon spp. by three plant meals

Based on biochemical assays and electrophoretical methods, the inhibitory effects of three plant meals (soybean meal, wheat meal, winged bean meal) on digestive alkaline proteases of discus were investigated. Casein assays revealed that increasing levels of soybean meal caused a linear inhibitory effect on activity of protease. SDS-PAGE images revealed that trypsin and chymotrypsin were the inhibited enzymes. Soybean showed the lowest inhibition rate followed by wheat meal and raw winged bean. There was a quadratic relationship between wheat meal levels and its inhibition of protease acitivity. The highest inhibitory effect was obtained with the winged bean meal with inhibition of caseinolytic activities ranging from 3.6–98.6%. Results from this study showed the potential of both soybean meal and wheat meal as ingredients for practical diet for discus, while demonstrating the need for further improvement in processing method for winged bean meal.     

The effects of variation in feed protein level on the culture of white shrimp, Litopenaeus vannamei (Boone) in low-water exchange experimental ponds

The present study was conducted to evaluate the effect of varying dietary protein level on pond water quality and production parameters of white shrimp Litopenaeus vannamei (Boone). Experimental units consisted of nine 400‐m² earthen ponds with a low water exchange. Two treatments were tested: treatment HP consisted of shrimp fed a high‐protein diet (40%) during the whole grow‐out, and treatment LP consisted of the use of a low‐protein diet for the complete farming period. No differences on any of the water quality parameters were observed among treatments. Excellent survival (over 85%) and feed conversion ratios (around 1.6), and acceptable growth (over 12 g) and biomass (from 1721 to 1793 kg ha^?1) were recorded in all experimental ponds. No significant differences in any of the production parameters were found among treatment groups.     

Strategies for further development of aquatic feeds

ABSTRACT: To date, aquatic feeds have depended heavily on fishmeal and fish oil as their source of protein and lipid. However, the feed industry is encountering shortfalls in the availability of these ingredients because of a decline in the number of fish captured in the wild and the increased human demand for some of the species currently being used for fishmeal and oil production. Therefore, efforts are now being directed in different parts of the world to finding alternative quality ingredients, which ideally are less expensive and readily available for use in practical diets. The data accrued have shown that a large proportion of both fishmeal and fish oil can be replaced by other protein and lipid sources. However, it is emphasized that an optimal essential amino acid balance be maintained and that the n-3 highly unsaturated fatty acid requirement be satisfied when combining economical protein and lipid ingredients. Newly developed feeds should aim at being nutrient-dense in order to reduce the output of solid, P and N waste. This can be done through improving nutrient availability, optimizing the digestible protein to energy balance of the diet, and replacing dietary fishmeal with alternate ingredients. These diets should also be effective for maintaining good health and improving disease resistance in fish through enhancement of immune responses. A wholesome approach to culturing fish would be to use appropriate feeding standards that are aimed not only at improving economic returns but also at developing a lasting cohabitation of sustainable aquaculture and a cleaner environment. Furthermore, in the 2lst century aquaculture would still retain its place as a prominent source of food protein, signifying that fish feed research remains a forceful area in discovering better feeds for the industry.     

Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans

The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.     

Allozyme diversity in Australian rainbow trout, Oncorhynchus mykiss

Rainbow trout, Oncorhynchus mykiss (Walbaum), were first introduced into Australia over 100 years ago, and forms the basis of important recreational inland fisheries and an aquaculture industry in south‐eastern Australia. This paper investigates the genetic variation within and between samples of Australian rainbow trout using allozyme electrophoresis. The levels of genetic diversity within Australia do not show marked differences from those observed in hatchery and wild populations from throughout North America, New Zealand and South Africa, but there is evidence for the loss of some rare alleles during translocation from California to Australia via New Zealand. No appreciable difference in genetic diversity was apparent between hatchery and self‐sustaining wild populations of rainbow trout from mainland Australia. However, significant differences in allelic frequencies were observed, with consistent genetic differences between Victorian and New South Wales samples most likely reflecting state‐based hatchery and stocking policies.     

稀有白甲鱼含肉率及肌肉营养成分分析

试验结果表明，稀有白甲鱼含肉率为69．92％；肌肉中蛋白质、脂肪和灰分含量分别为16．58％、2．05％、0．99％；肌肉中17种常见氨基酸（除色氨酸）总含量（占肌肉干重）为77．89％。其中必需氨基酸含量为36．27％，占氨基酸总含量的46．56％。必需氨基酸与非必需氨基酸含量的比值为0．87。鲜味氨基酸含量为26．26％，占氨基酸总含量的33．70％。必需氨基酸指数为70．44。缬氨酸和异亮氨酸分别为第一、二限制性氨基酸。     

Dietary plant protein utilization in Atlantic cod, Gadus morhua L.

The present trials aimed to investigate the effects of replacing fish meal with plant proteins in diets for cod, using a regression design where fish meal constituted the control. The plant protein diets were formulated to meet the amino acid requirements according to NRC (1993) and contained corn gluten meal, soybean meal, a mixture of these, or a mixture of wheat gluten meal and soybean concentrate. The plant protein fraction constituted up to 440 g kg⁻¹ of the extruded diet. Two feeding experiments were conducted, one at high (11 °C) and one at low (6.5 °C) temperature. High growth and feed utilization were obtained in all diet groups at both temperatures. However, only in Exp.1 (11 °C), growth and feed utilization were linearly reduced by increases in dietary soybean and corn gluten meal, while no such effect was detected in Exp.2 (6.5 °C). Reductions in protein retention were seen at both temperatures with use of all evaluated plant protein sources. Due to large amounts of plant protein ingredients in diet, differences in diet amino acid composition were seen. These were partly reflected in the muscle free amino acid pool. Dietary plant ingredients did not affect whole body, liver or muscle proximate compositions, or liver indices. In both experiments, blood parameters were within the range of earlier reported normal values and indicated, together with low mortality, good fish health status. The results show that there is high potential to use protein‐rich plant ingredients in diets for Atlantic cod.