Determination of Salt Diffusion Coefficient in Rainbow Trout (Oncorhynchus mykiss)

ABSTRACT

The diffusion properties of sodium chloride in rainbow trout muscle during dry salting at 10°C were investigated. The average initial dry matter content of the samples increased from 27.80 ± 0.10 to 36.01 ± 0.16%, and the average salt content increased to 17.98 ± 0.38 g NaCl/100 g dry matter at the end of the 10-h dry salting process. The analytical solution of Fick’s second law considering one-dimensional diffusion through an infinite slab was used to calculate effective salt diffusion coefficients (D_eff). The D_eff values in rainbow trout fillets decreased with increasing salting time and ranged from 6.64 × 10^?10 to 16.45 × 10^?10 m²/s.     

虹鳟IL-17多克隆抗体的制备及初步鉴定

本研究根据NCBI已发表序列设计引物,提取经植物血凝素刺激后的虹鳟Oncorhynchus mykiss头肾细胞总RNA,采用RT-PCR方法扩增IL-17成熟肽基因,测序结果表明所获得的序列与发表序列相一致。将该基因重组至原核表达载体pET32a中,并转化大肠杆菌Escherichia coli Rosetta,进行诱导表达。SDS-PAGE电泳结果表明:目的蛋白以包涵体形式表达,大小约为32k Da,重组蛋白经Ni-NTA系统纯化、复性后纯度达90%以上,以其免疫小鼠制备虹鳟IL-17多克隆抗体。ELISA结果显示其效价为1∶25 600,而间接免疫荧光结果显示所制备的多克隆抗体能够特异性地识别真核细胞中瞬时表达的虹鳟IL-17。本研究成功制备虹鳟IL-17多克隆抗体,为下一步IL-17在虹鳟黏膜免疫中的作用及其佐剂效应研究奠定基础。     

Influence of Dietary Lipid and Protein Sources on the Sensory Quality of Organic Rainbow Trout (Oncorhynchus mykiss) After Ice Storage

The influence of dietary protein and lipid sources on the quality of organic rainbow trout (Oncorhynchus mykiss) was studied. The protein and oil sources were fishmeal, fish oil, and organic vegetable protein and oils. Sensory profiling was performed during 3 to 14 days of ice storage along with lipid analyses of the fillet. Overall, the results showed that the sensory characteristics of the trout were affected in different ways during ice storage. The source of lipid seemed to affect the sensory quality at the beginning of the storage period, while the protein source seemed to have a more pronounced impact at the end of the storage period.     

二倍体与三倍体虹鳟(Oncorhynchus mykiss)红细胞及部分血液指标的比较

对三倍体和二倍体虹鳟(Oncorhynchus mykiss)的红细胞大小(长、短径值和体积)、形态及部分血液指标(红细胞数、血红蛋白、红细胞压积、红细胞脆性和白细胞数)进行了比较。结果表明:三倍体虹鳟红细胞体积与二倍体比值为1.51∶1,细胞核大小与二倍体比值为1.49∶1,与理论值(1.5∶1)比较均无显著差异(P>0.05)。二倍体红细胞短径/长径比值为0.63,呈现出椭圆形;三倍体红细胞短径/长径比值为0.60,呈现出明显的长椭圆形,并有19%的红细胞有哑铃形或分裂的细胞核,这可作为区别二倍体鱼类的一个重要鉴定标志。三倍体虹鳟的红细胞数、红细胞压积和红细胞脆性明显小于二倍体(P<0.05),而血红蛋白与白细胞数与二倍体比较无明显差异(P>0.05)。     

二倍体和三倍体虹鳟精液低温保存寿命的研究

与二倍体（2n组）精原液做比较,以不同密度为组别,研究了4℃下两组（3n-HD和3n-LD）三倍体虹鳟（Oncorhynchus mykiss）精原液的保存时限。结果发现,随着保存时间的延长,精液中精子的活力和寿命逐渐降低。2n组、3n-HD组和3n-LD组虹鳟精原液中精子分别在保存5d、4.5d和12.5d后失去活力;在7d、15d和14.5 d后死亡,即三倍体虹鳟精原液4℃保存时间约为二倍体的2倍,说明低温短期保存三倍体虹鳟精原液可以取得较好效果。     

Prediction of the Weight of Aquacultured Rainbow Trout (Oncorhynchus mykiss) by Image Analysis

Prediction relationships between weight and image features were established with a high correlation for whole aquacultured rainbow trout. Three hundred fish from three different farms were used. The fish were temporarily removed from the raceway, anesthetized, and their picture was taken by a digital camera. A reference square of known surface area and color was placed beside the fish. The fish were then returned to the raceway alive. The image was analyzed, and the view area of the fish was calculated using the area of the reference square. The average color of the fish was also determined (L*, a*, and b* values). The following equations were used to fit the view area (X) vs. weight (Y) data: linear, power, and second order polynomial. The R² values for the used equations were: linear = 0.98; power = 0.99; polynomial = 0.98. Image analysis can be used reliably to predict the weight of whole aquacultured rainbow trout. In addition, color and other visual attributes can be objectively determined by image analysis to sort by visual quality.     

二倍体和三倍体虹鳟精液低温保存寿命的研究

与二倍体(2n组)精原液做比较,以不同密度为组别,研究了4℃下两组(3n-HD和3n-LD)三倍体虹鳟(Oncorhynchus mykiss)精原液的保存时限。结果发现,随着保存时间的延长,精液中精子的活力和寿命逐渐降低。2n组、3n-HD组和3n-LD组虹鳟精原液中精子分别在保存5d、4.5d和12.5d后失去活力;在7d、15d和14.5 d后死亡,即三倍体虹鳟精原液4℃保存时间约为二倍体的2倍,说明低温短期保存三倍体虹鳟精原液可以取得较好效果。     

荧光染色法检测虹鳟精子质量

本实验运用三种荧光染色方法对虹鳟精子的质量进行检测,并探讨荧光染色对虹鳟精子质量评价的可性及意义。在生殖季节采集虹鳟精液,分别进行Hoechst33342/碘化丙啶双染,吖啶橙染色及罗丹明123/碘化丙啶双染,然后将其制成涂片荧光显微镜下观察。经荧光染色的虹鳟精子质膜完整,DNA完整,可明显分辨出活体和死亡精子,线粒体膜功能正常精子的比率分别为90.7%、98.7%和96.4%。综上所述,结合三种检测方法可以简便快速地测定虹鳟精子的质量。     

新型锌制剂对虹鳟生长及免疫功能的影响

为研究纳米锌及纳米锌多糖复合体对虹鳟生长及免疫功能的影响,选取平均体质量(120.0±3.0) g的虹鳟300尾,随机分成5组,每组3个平行,每个平行20尾,按1μL/g进行尾部肌肉注射纳米锌及纳米锌多糖复合体溶液,含量分别为1000、3000 mg/kg,试验周期为10 d。试验结果显示,纳米锌及纳米锌多糖复合体可不同程度提高虹鳟特定生长率(P>0.05)。1000 mg/kg纳米锌多糖复合体组虹鳟血液NBT阳性细胞数量百分比在第3 d最高,达22.3%(P<0.05);3000 mg/kg纳米锌多糖复合体组白细胞吞噬能力在第6 d最强(P<0.05)。3000 mg/kg纳米锌组虹鳟血清杀菌能力第3 d最高(19.4%),而3000 mg/kg纳米锌多糖复合体组血清杀菌能力第6 d最高,达32.43%(P<0.05)。第3 d,1000 mg/kg纳米锌多糖复合体组过氧化氢酶活性、一氧化氮合成酶活性分别为26.39、43.38 U/mL (P<0.05);第6 d,超氧化物歧化酶、酸性磷酸酶活性最高,分别为29.91 U/mL、201.4 U/L(P<0.05),微量丙二醛浓度最低,为3.19 nmol/mL。试验结束后通过杀鲑气单胞菌攻毒感染,7 d内观察虹鳟累积死亡率,1000 mg/kg纳米锌多糖复合体组累积死亡率仅30%。由试验结果可知,纳米锌和纳米锌多糖复合体均能对虹鳟免疫功能产生促进作用,以1000 mg/kg纳米锌多糖复合体效果最佳。     

植酸酶对金鳟粪磷及水环境中磷含量的影响

通过喷涂和预处理两种方法将不同浓度植酸酶添加到豆粕型金鳟(Oncorhynchus mykissW albaum)饲料中,研究了植酸酶对金鳟粪磷及养殖水环境中磷含量的影响。金鳟粪磷分析结果显示,喷涂法添加植酸酶,对照组中粪磷含量最高,其他各试验组较对照组差异显著(P<0.05),但500~2000 U/kg组中粪磷含量差异显著(P<0.05),2500~6000 U/kg组中粪磷含量无显著差异(P(0.05);预处理法添加植酸酶,对照组粪磷含量最高,其他各试验组粪磷含量较对照组降低显著(P<0.05),但添加植酸酶的各试验组之间差异不显著(P(0.05)。通过水中磷含量试分析结果显示,无论是喷涂法添加植酸酶还是预处理法添加植酸酶,各试验组水体总磷含量分别较其对照组差异不显著(P(0.05),各试验组的活性磷含量均未检出。实验结果表明,金鳟饲料中添加植酸酶可以降低粪磷含量,对养殖生产过程中控制磷的排放,减少磷污染具有重要的意义。     

Use of Underground Brackish Water for Reproduction and Larviculture of Rainbow Trout,Oncorhynchus mykiss

Broodfish were reared for 6 months in underground brackish water (11 ppt), and the quality of eggs and semen was evaluated in comparison to broodfish held in fresh water. Fry produced by the broodfish in brackish water (S) and freshwater (F) were then reared in different salinities (1, 4, 7, and 10 ppt). Broodfish successfully matured and spawned in underground brackish water, and gonad quality was better than of the broodfish in fresh water (p < 0.01). Growth indices of fry produced by both groups decreased with increasing salinity (p < 0.01). Survival was not affected by the salinities tested. Underground brackish water can be used as a source to induce successful sexual maturation and to produce high-quality gonads.     

虹鳟营养需求的研究进展

简要介绍了近年来国内外虹鳟的营养研究,包括蛋白质、脂肪、维生素、矿物质等方面,提出一些虹鳟养殖中的注意事项,为虹鳟养殖、饲料生产和科研开发提供参考。     

Dietary essential amino acids and heat increment in rainbow trout (Oncorhynchus mykiss)

Oxygen consumption attributable to apparent heat increment (AHI) was measured in relation to varying essential amino acid proportions (EAA) infused into rainbow trout,Oncorhynchus mykiss (250–450 g), induced to swim at 1 BL s^–1. Five diets, mimicking EAA concentrations in trout whole body protein, deficient in the branched chain amino acids (isoleucine, leucine and valine), containing unbalanced proportions of EAAs and supplying lysine in excessive and limiting proportions, were tested. Following infusion of the experimental diets, a significant increase in oxygen consumption was observed. Changes in plasma EAAs following infusion paralleled the time course of AHI (i.e., oxygen consumption). AHI represented the equivalent of 15–32% of the gross energy intake depending on dietary EAA composition. Diets supplying EAAs similar to trout whole body protein and limiting in lysine produced the lowest AHI values, indicating efficient utilization of dietary amino acids. Higher AHI values were associated with diets deficient in the branched chain amino acids and diets supplying lysine in excess. Duration of elevated metabolism was independent of both dietary composition and energy intake. Different proportions of EAAs in the diet can increase the energy expended as AHI. In an attempt to reduce the energy liberated as AHI, attention must be paid to the quality, quantity and balance of dietary EAAs.     

Changes of Non-Specific Immune Parameters in Rainbow Trout,Oncorhynchus mykiss,After Exposure to Antimicrobial Agents Used in Aquaculture

Changes of plasma lysozyme and serum bactericidal activity, ceruloplasmin, total plasma protein, serum glucose, hematocrit and leucocrit levels in rainbow trout (Oncorhynchus mykiss) were studied after exposure to formalin and chloramine-T, which are used in aquaculture as anti-ectoparasitic and antibacterial agents. Fish were exposed to formalin at a concentration of 250 ppm for 60 min and exposed to chloramine-T for 5 ppm for 3 h. The parameters were analyzed for rainbow trout exposed to formalin and chloramine-T soon after treatments and rainbow trout exposed to freshwater after 24 h (return) following the treatments. Plasma lysozyme activity decreased after exposure to formalin; however, it was not affected by chloramine-T exposure. Serum bactericidal activity reduced after both formalin and chloramine-T exposure and did not change after returning to freshwater. Ceruloplasmin and total plasma protein were not affected by both treatments. Serum glucose levels of exposed fish elevated and remained at higher levels during recovery period when compared to control values. Hematocrit values of fish exposed to formalin increased; however, no significant change was observed in fish exposed to chloramine-T. Leucocrit levels of fish treated with both formalin and chloramine-T were higher than that of controls.     

Effect of feeding on the tissue free amino acid concentrations in rainbow trout (Oncorhynchus mykiss Walbaum)

This study investigates whether tissue free amino acid (FAA) pools in rainbow trout,Oncorhynchus mykiss (Walbaum), are altered following feeding and the relationships between the amount of food consumed and the FAA pool size. Trout were starved for 7 days to provide baseline data and then refed on day 8. Individual food intake was measured by radiography and the consumption of amino acids (AA) calculated from dietary protein consumption. Total FAA concentrations in the stomach, liver and white muscle were little changed at various times after the meal and this pattern was repeated for the majority of individual FAA. Overall, the most notable change was a reduction in essential FAA concentrations (principally in valine, leucine and isoleucine) in the white muscle following feeding. However, in the caeca total FAA, total essential FAA and a number of individual FAA were significantly elevated at 4, 9 and 15h following feeding. There were few significant correlations between dietary amino acid consumption and total tissue FAA and essential FAA concentration in the stomach, caecum and white muscle; correlations were stronger in the liver. In order to explain the relative constancy of total FAA concentrations in the tissues following food intake (which represents over 100% of the total FAA pool) a model is presented that quantifies the AA flux through the free pools and considers the role of protein turnover in regulating FAA pool size.     

植酸酶对金鳟体成分及金鳟椎骨中钙、磷含量的影响

采用喷涂法和预处理两种方法,在豆粕型金鳟饲料中添加不同浓度的植酸酶,通过90 d的饲喂试验,研究植酸酶对金鳟(Oncorhynchus mykiss Walbaum)体成分及其椎骨中钙、磷含量的影响。结果显示:喷涂法和预处理法添加植酸酶的处理组与对照组相比,粗蛋白、粗脂肪的含量均有所下降;喷涂法添加植酸酶的处理组的鱼体粗灰分、磷和钙含量均高于对照组,其中,喷涂法添加1500 U/Kg植酸酶组鱼体磷含量、灰分含量和钙含量显著高于对照组(P(0.05),且此组鱼体磷高于其它试验组;喷涂法各组椎骨中粗灰分、磷含量较对照组提高显著(P(0.05),2500 U/kg植酸酶组椎骨磷含量最高,椎骨中钙含量除D1和D8组外,添加植酸酶的处理组均较对照组提高显著(P(0.05)。预处理法添加植酸酶的处理组与对照组相比,鱼体和椎骨中粗灰分、磷和钙含量均有显著提高(P(0.05),但植酸酶试验组间椎骨中粗灰分、磷含量和钙含量差异均不显著(P(0.05)。结果表明,金鳟饲料喷涂法添加植酸酶1500~2500 U/kg浓度最适,预处理法添加植酸酶500 U/kg浓度较为宜。     

Sox9和amh基因在雌、雄虹鳟和伪雄鱼各组织中的表达模式分析

本研究利用Real-timePCR分析和比较sox9与amh基因在雄、雌虹鳟Oncorhynchus mykiss及及其伪雄鱼各组织中的表达模式。结果表明:sox9基因在这三种鱼性腺、脑、脾、肝以及脑垂体中的表达显著高于其他组织(P<0.05)。在性腺组织中,sox9基因在雄鱼中的表达量显著高于伪雄鱼(P<0.01),而在伪雄鱼中的表达量又显著高于普通雌鱼(P<0.01)。脑组织中,雌鱼sox9的表达显著高于雄鱼和伪雄鱼(P<0.05)。Amh基因在性腺和脑组织中的表达显著高于其他组织(P<0.05)。该基因在伪雄鱼性腺和脑中的表达均显著高于雄鱼和雌鱼(P<0.05)。结果表明,外源雄激素的诱导可导致伪雄虹鳟性腺和脑组织中sox9和amh基因的表达显著高于雌性虹鳟,促进其性腺的雄性化发育。本研究可为全雌性虹鳟制种技术研究奠定重要理论基础。     

Dynamics of plasma free amino acids in rainbow trout (Oncorhynchus mykiss under variety of dietary conditions

Plasma amino acid profiles in the systemic circulation of rainbow trout acclimated to diets containing different protein sources, with and without supplementation with free amino acids, were monitored for up to 120 hours after consumption of the diets. Plasma concentrations of lysine, methionine, and isoleucine increased more rapidly after feeding a diet supplemented with these amino acids in free form and, depending upon the increase in plasma concentration, remained at concentrations above those in fish fed the basal diet for at least 24h after feeding. Dietary supplementation with isoleucine increased plasma concentrations of leucine and valine as well as of isoleucine. Maximum plasma concentrations for most amino acids were attained between 12 and 24h postprandial. Dietary inclusion of gelatin caused more rapid elevations in plasma glycine and serine than did free glycine. Feeding at three hour intervals resulted in stable plasma concentrations of individual amino acids in contrast to the fluctuations occurring when fish were fed once daily. Frequent feeding favoured a higher proportion of protein to lipid in body composition.     

热休克诱导全雌虹鳟三倍体

以虹鳟二倍体的伪雄鱼为父本（遗传型为xx），探讨了采用热休克方法阻止第二极体排放诱导全雌虹鳟三倍体的适宜条件。结果表明：虹鳟三倍体诱导率明显受处理温度、起始时间以及持续时间等因子的影响。在孵化水温为6．5℃，卵子受精后20min经26℃热处理20min，孵化率为64．62％，三倍体诱导率为86．67％；卵子受精后lOmin经26℃热处理20min，三倍体诱导率为100％，但孵化率仅为3．87％。受精卵经26℃处理的诱导效果好于24℃和28℃的（P〈0．01）。     

Effect of Dietary Safflower and Canola Oil on Growth Performance,Body, and Fatty Acid Composition of Rainbow Trout (Oncorhynchus mykiss)

The objective of the present study was to evaluate the effects of the diets enriched with safflower and canola oil on growth, feed utilization, body composition, liver, and muscle fatty acid composition of rainbow trout (Oncorhynchus mykiss). Rainbow trout having approximate initial weight of 97.03 ± 0.10 g were fed the experimental diets containing only fish oil (Group 0SFO), safflower oil (50% safflower oil, Group 50SFO and 33% safflower oil, Group 33SFO), and vegetable oil blend (33% safflower and 33% canola oil, Group 66SFCO) for 45 days. Twenty-five fish were randomly assigned for triplicate treatments and offered the test diets two times daily to apparent satiation. At the end of the experiment, survival was 100% in all treatments. No significant differences in the weight gain, specific growth rate, feed conversion ratio, and protein efficiency ratio were found between fish fed with the different experimental diets. The highest hepatosomatic index (HIS) and viscerasomatic index (VSI) was obtained in 50SFO and 33SFO groups, respectively. The moisture, protein, lipid, and ash content in the body composition of the fish increased in all experimental groups. The lipid content was not significantly different among the groups (p > 0.05); however, there was a significant difference in ash content between the control and the other groups (p < 0.05). The experimental diets containing vegetable oil (50SFO and 33SFO groups) and vegetable oil blend (66SFCO group) had significantly higher concentrations of n-6 fatty acids, predominantly in the form of linoleic acid (LA). The n-3 fatty acids were present in significantly higher concentration in the control treatment (0SFO). The fatty acid composition of fish fillet and liver were reflective of the dietary lipid source. While the fillet and liver of fish fed the 50SFO diet was high in linoleic acid (18:2 n-6), fish fed the 66SFCO diet had high concentrations of oleic acid (OA; 18:1 n-9). The present study suggests that fish oil can be replaced by up to 50% with safflower oil and by up to 66% with safflower + canola oil blend in rainbow trout diets with no significant effect on growth.