Development of a rainbow trout intestinal epithelial cell line and its response to lipopolysaccharide

A cell line, RTgutGC, was developed from the intestine of Oncorhynchus mykiss. RTgutGC has an epithelial‐like shape, been passaged over 100 times, and cryopreserved successfully. A rainbow trout origin was confirmed by sequencing a 652 bp region of the mitochondrial cytochrome c oxidase I gene. RTgutGC is grown routinely in Leibovitz’s L15 without glutamine supplemented with 10% fetal bovine serum (FBS). Cell viability was evaluated with Alamar blue (AB) for metabolic activity and carboxyfluorescein diacetate acetoxymethyl ester (CFDA AM) for membrane integrity. Viability was unchanged by lipopolysaccharide (LPS) for cultures in FBS. For cultures at low cell densities in L15 without FBS or glutamine, cell viability declined in a LPS dose‐dependent manner, allowing calculation of the concentration causing a 50% decline in viability (EC₅₀). When glutamine was present, the EC₅₀ was increased for both AB and CFDA AM. As the cell density increased, LPS became much less cytotoxic and no EC₅₀ could be calculated for very confluent cultures. Only high‐density cultures had alkaline phosphatase (AP) activity. Thus, glutamine and possibly AP protect against LPS cytotoxicity. RTgutGC should be a useful in vitro tool for studying problems of nutrition and gastrointestinal health in fish.     

Minimising aerobic respiratory demands could form the basis to sub-lethal copper tolerance by rainbow trout gill epithelial cells in vitro

Mechanisms of Cu tolerance were investigated in respiratory epithelial cell cultures, from rainbow trout gills, by studying O₂ consumption and protein synthesis rates, intracellular Na concentration and TER. The lowest concentration found to reduce O₂ consumption was 25 M Cu. This did not affect either protein synthesis rate or intracellular Na concentration and was interpreted in terms of copper tolerance; i.e., how these two energetically demanding processes are maintained despite a reduction in aerobic ATP supply. The relationship between protein synthesis rate and synthesis cost is exponential and the cost of protein synthesis in gill cells was found to be minimal (i.e., this cell occupies a position on the asymptotic section of the protein synthesis rate/synthesis cost model) and unaffected by 25 M Cu. Thus protein synthesis rates could be maintained since any reduction would represent an insignificant energy saving. Intracellular Na concentrations and O₂ consumption rates were linearly correlated suggesting reducing intracellular maintenance costs would have a greater significance in terms of overall energetic conservation. Intracellular Na maintenance costs, calculated from O₂ consumption rates and intracellular Na concentrations, were found to decline after exposure to 25 M Cu. Since TER was unaffected this implied the reduced costs arose from membrane `channel arrest'. Thus the Na/K ATPase energy demands, associated with maintaining intracellular Na concentration, could be reduced by decoupling metabolic demand and membrane function. Therefore this study may demonstrate how the flexibility of cellular energetics enables gill epithelial cells to tolerate sub-lethal Cu.     

Acclimation of rainbow trout to sea water

In some fish farms it is advantageous to transfer rainbow trout from fresh water to sea water at as small a size as possible. It has been shown that a relatively small reduction in salinity at the marine site can give appreciable improvements in survival rates during acclimation by smaller fish, thus rainbow trout of 15 g have routinely been put directly into sea cages at a salinity of 22% with mortality levels of from 1 to 8%.     

虹鳟含肉率及肌肉营养成分分析

测定了6尾虹鳟的含肉率及其中4尾肌肉的营养成分,并与部分淡水鱼进行了比较,对其营养价值作了综合评定。虹鳟的含肉率达75.61%。鱼肌肉粗蛋白质质量分数为21.11%,粗脂肪质量分数为3.53%。肌肉中18种氨基酸的总量为18.63%,其中8种必需氨基酸和4种鲜味氨基酸分别占8.37%和6.79%,必需氨基酸占总氨基酸量的44.92%,EAAI为69.20,低于鸡蛋蛋白质模式,与WHO/FAO模式基本相同。第1限制性氨基酸为蛋氨酸 胱氨酸,CS和AAS分别为0.358和0.672。     

Comparison of different European rainbow trout populations under intensive production conditions

Growth and carcass quality of European rainbow trout populations were compared in a full sib family test over five experimental periods. In total, 131 full-sib families of 17 different populations, (i.e. 19 500 fish), were included. A special system for family testing in trout was developed. This consisted of separate hatching of single pair matings, separate rearing of families up to marketable size and common fattening under two stocking densities. Thus, growth was studied separately for the rearing and the fattening period. The observed differences in rearing and fattening weight between and within populations were quite distinct and of significance for production and breeding efficiency. Similar ranking of populations after fattening in different years and under different stocking densities indicated that the final weight was highly genetically controlled. Carcass characteristics determined at marketable size showed somewhat smaller differences between populations. Populations with the highest fattening weight did not generally produce the most valuable carcasses. The results are discussed with respect to their impact on breeding strategies.     

Demand-feeding behaviour of rainbow trout

Rainbow trout learned quickly to operate a trigger to deliver dry food and showed a peak of feeding at dusk, but also appreciable nocturnal feeding. Food deliveries were aggregated into feeding bouts with a preferred time between trigger presses of 4–8 min; the aggregation of feeds was more marked in groups of trout than with trout held singly. Total daily food intake varied with the reward level per trigger press, and could therefore be controlled at about the manufacturers recommended feeding levels. Territorial and hierarchical behaviour were observed and in groups of up to 20 trout only one member of the group pressed the trigger although all the fish took the food delivered, and there was no evidence that smaller fish in a group were deprived of food to the point at which they lost condition.     

虹鳟养殖技术之一利用水库底层水养殖虹鳟

虹鳟鱼属冷水性鱼类,生长快,抗病力强,是目前深受养殖户和消费者喜爱的品种之一.2002年,我们利用凌源菩萨庙水库底层水进行虹鳟商品鱼养殖试验,取得较好的经济效益.     

Effect of varying dietary protein level in different families of rainbow trout

Groups of rainbow trout (Salmo gairdneri) fingerlings from five different families and five inbred groups were fed for 153 days on each of four diets similar in energy content, but differing in their percentage of protein. The protein contents were 24, 33, 42 and 51%, respectively. Significant differences between fish families were found for growth, condition factor, chemical composition of the carcass, and digestibility of protein. At the end of the experiment, interaction between diet fed and family was found close to significance for weight and relative growth rate, and significant for length and condition factor. This interaction indicates that prospects for selectively breeding strains of rainbow trout specifically better able to utilize protein are promising.Fish growth rate increased with increasing dietary protein content. The corresponding condition factor, content of dry matter and energy in the carcass decreased. The high protein diets gave more protein in the carcass and a more discoloured liver than the low protein diets. Apparent protein digestibility increased with increasing protein level.     

不同动植物蛋白比饲料对虹鳟生长的影响

虹鳟(Oncorhynchus mykiss)是一种名贵的鲑科鱼类,肉质细嫩,味道鲜美,无小骨刺、无需刮鳞,蛋白质和脂肪含量高,高度不饱和脂肪酸含量高于其他鱼类.虹鳟苗种的密养技术和商品鱼的高产养殖技术已在全国推广,目前已遍布全国20多个省.近几年来,虹鳟养殖在我省各县逐渐兴起,并取得了较好的经济效益.虹鳟是典型的肉食性鱼类,对它的营养需要已做了较详细的研究,在国外虹鳟配合饲料的使用已较普及,配方技术也较成熟,但是我国目前虹鳟饲料的配制技术水平与国外先进水平相比尚有一定差距.     

提高虹鳟、金鳟鱼苗成活率的关键技术

在虹鳟及金鳟鱼苗的养殖中,鱼苗的成活率是养鳟鱼场取得良好经济效益的关键,也是衡量鱼场技术水平的一个重要指标。根据多年的生产实践经验,总结出以下几点,供广大鳟鱼养殖户参考。     

Interaction of low density lipoproteins with liver cells in rainbow trout

Liver is the main catabolic tissue for low density lipoprotein in rainbow trout (Gjøen and Berg 1992). We have investigated the interaction of LDL with isolated trout liver cells and liver membranes. ¹²⁵I-TC labelled trout LDL bound to isolated trout liver cells in a time dependent and saturable manner with an apparant K_d of 20.1 g/ml, suggesting the existence of a specific binding site on the surface of these cells. The binding was Ca²⁺ dependent assessed by the 50% reduction obtained by 5 mM EDTA. Saturable binding to isolated trout liver membranes could also be demonstrated, but with lower affinity as compared to intact cells. Degradation of ¹²⁵I-TC-LDL in hepatocytes was also saturable as degradation could be inhibited about 60% by a 100 fold surplus of unlabelled LDL. The rate of degradation increased with temperature up to 20°C. Both cell association (binding + uptake) and degradation were reduced down to 20% of control in the presence of microtubular and lysosomal inhibitors. Hepatic catabolism of trout LDL therefore seems to depend on receptormediated endocytosis, followed by lysosomal degradation.Abbreviations TC tyramine cellobiose - LDL low density lipoproteins - MeLDL methylated low density lipoproteins - VLDL very low density lipoproteins - HDL high density lipoproteins - VTG vitellogenin - EDTA ethylenediamine tetraacetic acid - PBS phospate buffered saline - SDS-PAGE sodium dodecyl sulphatepolyacrylamide gel electrophoresis - DMPC L--phosphatidylcholine-dimyristoyl     

Primary responses of rainbow trout to dietary phosphorus concentrations

A series of experiments was conducted to research sensitive indicators for phosphorus status of fish using post‐juvenile rainbow trout. Fish were fed up to 24 days with semipurified diets that varied in phosphorus content. Concentrations of glucose‐6‐phosphate, ATP, creatine phosphate, glucose, total lipids and total cholesterol in blood or skeletal muscle were relatively unchanged by the 24 days of dietary phosphorus restriction. Inorganic phosphorus and ATP levels in the blood, however, correlated significantly and positively. Inorganic phosphorus levels in plasma and urine were significantly lower in fish fed phosphorus‐deficient diets than phosphorus‐supplemented diets. Urinary phosphorus excretion peaked 6–10 h after feeding fish with diets containing potassium phosphate. Fish receiving either commercial feeds or experimental diets containing phosphorus as fish bone excreted trace amounts of phosphorus in the urine. Faecal content of phosphorus significantly increased when the diet containing potassium phosphate was supplemented with calcium carbonate. Urinary phosphorus concentration was found to be a rapid and sensitive indicator for dietary intake of phosphorus and probably phosphorus status of the fish, and had an advantage over conventional response variables in estimating dietary phosphorus requirement especially with large commercial‐size fish.     

Attachment of Vibrio pathogens to cells of rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract. The attachment of Vibrio pathogens to cells of rainbow trout, Oncorhynchus mykiss (Walbaum), was studied by use of species-specific monoclonal antibodies and indirect FITC- immunofluorescence microscopy. Vibrio anguillarum, V. ordalii and V. parahaemolyticus attached to cultured cells of rainbow trout gonads, various tissues of cryostat sections of whole fish, and smears of gills, intestine, buccal mucosa and skin. The attachment was inhibited by prior incubation of bacteria with monoclonal antibodies at titres of 1:32, or less. Other Vibrio pathogens used in this study did not attach to any trout cells. The research provides approaches to study the mechanisms of bacterial attachment in the onset of Vibrio infections.     

虹鳟养殖技术 之三武陵山区养殖虹鳟应注意的技术环节

虹鳟是典型的冷水性鱼类,2005年北京市对口支援湖北省巴东县虹鳟鱼养殖项目,当年5月引进鱼苗1.5万尾后放入引水水渠中饲养,之后在水渠下方建新鱼池,并将鱼苗转入新池     

Progressive transfer to seawater enhances intestinal and branchial Na+-K+-ATPase activity in non-anadromous rainbow trout

Two sizes of rainbow trout [170 (4), 91 (3) g] were progressively transferred to seawater in January 1995 using three steps of 9, 20 and 28. Na+-K+-ATPase activity in gills, intestine and kidney, and plasma sodium and magnesium levels were assessed in response to changed external salinity. Gradual transfer to seawater had a stimulatory effect on gill and intestinal Na+-K+-ATPase activities after the transfer to 28, while its activity remained unchanged in the kidney. Plasma sodium content was not modified, while magnesium levels increased in response to increased external salinity. The size-dependent response to seawater transfer described by other authors was not detected in our experiments. The results are discussed in terms of long-term adaptation to seawater.     

Distribution of adipocyte-related cells in skeletal muscle of rainbow trout Oncorhynchus mykiss

Adiponectin is one of the adipokines secreted mainly from adipocytes in mammals. In rainbow trout, however, adiponectin is highly expressed in skeletal muscle. Although it has been suggested that fish skeletal muscle contains adipocytes, their endocrine function and distribution are poorly understood. Recently, an EST analysis of rainbow trout found that heart-type fatty acid binding protein (H-FABP), a member of the intracellular fatty acid binding protein family, encodes an adipose-specific gene. In this study, we produced anti-adiponectin and H-FABP antibodies and investigated the distribution of adipocytes and related cells in skeletal muscle of rainbow trout. The adiponectin signal was detected at around 75 kDa in muscle in Western blotting. Since the molecular mass of rainbow trout adiponectin is around 25 kDa, this 75 kDa band would be a trimer. For H-FABP, the signal band was detected at around 15 kDa. Immunohistochemistry of skeletal muscle sections indicated that adiponectin and H-FABP signals were present outside of muscle cells and throughout the muscle tissues, suggesting the existence of adipocyte-related cells in these regions. These results will contribute to our understanding of energy metabolism in fish skeletal muscle.     

应用不同模型估计虹鳟生长性状的遗传参数

利用黑龙江水产研究所渤海冷水性鱼类试验站2001至2006年5个品系虹鳟生长性状(体重、体长和肥满度)的测定数据,应用单性状和两性状动物模型估计遗传参数并对方差组分的差异比较分析.结果如下:体重的遗传力为0.20±0.05～0.45±0.03、体长为0.27±0.04～0.60±001、肥满度为0.32±0.06～0.47±0.03.虹鳟12月龄的生长性状间存在着较强的表型相关和遗传相关,其中表型相关为0.92、0.84、0.88,遗传相关为0.92±0.03、0.90±0.05、0.92±0.02;18月龄生长性状的遗传和表型相关分别为0.53±0.01、0.51±0.05～0.43±0.02、0.22、0.06～0.22;24月龄生长性状的遗传和表型相关分别为0.31±0.04、-0.91±0.14、-0.90±0.11、0.36、-0.03、-0.13;从不同模型的比较结果来看,体长和肥满度性状均以用两性状动物模型检测效果较好,体重则表现为单性状动物模型强于两性状动物模型.     

Phagocytic cells in the epidermis of rainbow trout, Salmo gairdneri Richardson

Abstract. Electron microscopical observations of skin of rainbow trout revealed the presence of several types of phagocytic cells in the epidermis, most notably macrophages, some of which were seen crossing the basal lamina. Occasionally, filament-containing cells showed either melanin granules or phagosomes, suggesting these cells were also capable of phagocytosis. No cells with the characteristics of the Langerhans cells were observed, which does not necessarily preclude the possibility of macrophages being able to play an identical role as antigen-trapping cells.     

Production of all-female triploid rainbow trout

Triploid rainbow trout were produced by heat shock treatment of eggs soon after fertilization with either normal sperm or sperm from masculinised females. The proportion of triploid fry, as judged from red blood cell nuclear volume, varied between 75% and 100% in three experiments using different batches of eggs from an autumn-spawning strain of trout while a single batch of eggs from a winter-spawning strain yielded 50% triploids.

A microscopic examination of the gonads was made on 5-month-old fish weighing between 1 and 3 g. In female controls the ovaries were packed with oocytes while those from female triploids, although showing the typical lamellar structure of an ovary, contained no oocytes, thus indicating that female triploids are sterile. The testes from triploid males appeared to be developing normally.

The use of masculinized females combined with heat shock treatment of eggs to produce triploids, allowed the production of sterile all-female triploids. This should have considerable potential for aquaculture.