Distribution of lipid droplets is an indicator for egg quality in brown trout, Salmo trutta fario

In the eggs of the brown trout, Salmo trutta fario, lipid droplets are clearly visible beneath the oolemma on the surface of the protein yolk. When examining the distribution of lipid vesicles in egg batches of different quality, four categories of lipid vesicle distribution could be distinguished: In the first category, the lipid droplets were evenly distributed throughout the egg. These egg batches had a high quality (percentage of eggs developing to eyed stage embryos: 96.2 ± 5.2). Some lipid droplets had the tendency to coalesce in one pole of the egg in the second category. This coalescing of lipid droplets was accompanied with a reduction in egg quality to about 50% of the first category (% of eyed stage embryos: 40.6 ± 15.2). In the last two categories, the lipid droplets were mostly coalesced in one or two poles of the egg and the egg quality was very low (% of eyed stage embryos: 3.3 ± 2.9 and zero, respectively). In conclusion, examination of lipid droplets distribution, in brown trout, can distinguish between high and low quality eggs in an easy and reliable way.     

Susceptibility by amberjack (Seriola dumerili), yellowtail (S. quinqueradiata) and Japanese flounder (Paralichthys olivaceus) to Neobenedenia girellae (Monogenea) infection and their acquired protection

This study investigated: 1) susceptibility differences to infection by Neobenedenia girellae (Capsalidae) between amberjack Seriola dumerili (Carangidae), yellowtail S. quinqueradiata and Japanese flounder Paralichthys olivaceus (Paralichthyidae); 2) growth and egg production of N. girellae on each fish species; 3) acquired protection of each fish species against this parasite. The number of N. girellae on S. dumerili was significantly higher than on S. quinqueradiata and P. olivaceus when these fishes were exposed to oncomiracidia in the same aquarium. Neobenedenia girellae growth on S. dumerili was fastest and, thus the number of eggs laid by parasites on S. dumerili was greater than on the other two species. Seriola dumerili and P. olivaceus, which were previously infected with N. girellae and treated by freshwater bath, acquired partial protection against re-infection by N. girellae. The relative re-infection of three S. dumerili individuals out of eleven individuals was markedly low compared with the initial infection, and the relative initial infection and re-infection on two P. olivaceus out of eleven individuals was markedly low. The results of this study could be useful to control N. girellae infections when cultivating S. dumerili, S. quinqueradiata and P. olivaceus.     

Acoustic conditioning for recall/recapture of escaped Atlantic salmon and rainbow trout

Escape of salmon from sea cages is a problem that continues to plague the aquaculture industry. Data collected during the past 15 years from Norway, Scotland, Ireland, Canada and U.S. suggest significant impacts on natural runs of fish and economic losses to producers. The present report investigated the feasibility of using acoustic conditioning as a means of recalling/recapturing escaped fish. Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) were found to respond to frequencies in the range of 50 - 400 Hz equally well. Subsequently, both species were conditioned to a 250 Hz acoustic tone during feeding. Juvenile and sub-adult fish readily conditioned to the acoustic signal within four days, with the maximum number of fish responding (85% salmon, 96% trout) by day seven. To assess retention of conditioning, fish were exposed to a single tone without feed reinforcement every one, two or four weeks. Salmon and trout continued to respond for a seven month period with no significant decrease (88% salmon, 97% trout) in response. No significant differences were observed in the response of either species to tones differing in frequency by up to 200 Hz (89% salmon, 96% trout) and intensity by 20 dB (91% salmon, 96% trout). Both species were reproducibly recalled to a cage or feeding ring in a 3.7 m tank, but were reluctant to re-enter the cage. The findings indicate that salmon and trout are readily conditioned to acoustic signals and retain that conditioning for an extended period of time without reinforcement. These characteristics suggest that acoustic conditioning has potential as a means to recall escaped salmon and when coupled with recapture, can reduce interactions with wild stocks and losses to the producer.     

Effects of diets containing a bacterial protein meal on growth and feed utilisation in rainbow trout (Oncorhynchus mykiss)

Triplicate groups of rainbow trout with initial weight 361 g were fed either a fish meal based control diet or diets containing 9, 18 or 27% bacterial protein meal (BPM) or 9% of an autolysate (AU) of the BPM. No significant treatment effects were found on specific growth rates (SGR), feed intake, feed efficiency ratio (FER), or retention of nitrogen, amino acids or energy. The apparent digestibility coefficients (ADC) of nitrogen, energy and most indispensable amino acids decreased when BPM was included in the diet. The ADC of lipid, sum of amino acids, arginine, lysine, threonine and most of the dispensable amino acids were reduced at 27% BPM inclusion compared to the control, 9% and 18% BPM diets. None of the ADC estimated was different in the 9% AU diet compared to the 9% BPM diet. The loss of nitrogen and energy in faeces per kg gain increased as the dietary BPM or AU levels increased, and the energy used for activity and maintenance was higher in fish fed the 27% BPM diet than in fish fed the other diets (P < 0.05). There were no significant differences in the urea concentrations in plasma, liver and muscle, whereas the uric acid level in plasma was elevated in trout fed the 27% BPM diet. Histological evaluation of tissue from the stomach, pyloric caeca, mid-intestine and distal intestine did not reveal any diet-related morphological changes.In conclusion, no significant differences in growth and feed efficiency were found in the rainbow trout fed diets containing up to 27% BPM, and the AU did not increase fish performance compared to the BPM. Based on the data from this study, at levels up to 27% dietary inclusion, BPM is a good replacement for fish meal in diets for rainbow trout.     

Metazoan parasites on gills of Southern Bluefin Tuna (Thunnus maccoyii) do not rapidly proliferate after transfer to sea cages

A relatively new and highly valuable aquaculture industry focuses on three species of bluefin tunas, which are captured from the wild and fattened for several months in sea cages. In teleost aquaculture, mortalities and extra production costs are very commonly associated with metazoan ectoparasites. In tuna, however, the production value lost due to diseases associated with ectoparasites is unknown. We collected epidemiological data on burdens of metazoans on the gills of farmed southern bluefin, Thunnus maccoyii, in a series of monthly samples of tuna from the time of stocking through to harvest (March to August, 2004; N = 210) in five sea cages on a farm off Port Lincoln, Australia. Three species were recorded; for one (a copepod, Pseudocycnus appendiculatus), there was a gradual, significant increase in both abundance (from a mean of 0.1 in March, to 3.83 in August) and prevalence (from 10% to 67.5%). For the other two species (a second copepod, Euryphorus brachypterus, and a polyopisthocotylean flatworm, Hexostoma thynni) there were no discernible trends in prevalences and abundances. These results contrast markedly with those of other intensively cultured species of finfishes, in which parasite epizootics are frequent. This finding may indicate that despite the stresses of captivity, tuna mount a robust immune response to ectoparasites; the relatively low stocking densities at which tuna are farmed may facilitate this. The fall in water temperature during farming (22 °C to 13 °C) may also reduce the reproductive rate of these ectoparasites.     

Isolation and partial characterization of rainbow trout (Oncorhynchus mykiss) gill mucin

Gill mucin from rainbow trout was isolated utilizing two rounds of cesium chloride density ultracentrifugation followed by gel filtration on Sepharose CL-2B. Neither density ultracentrifugation nor gel filtration alone was sufficient for purification of the mucin. Isolated gill mucin had a density of 1.5 g/ml and eluted at the void volume of the Sepharose CL-2B column. Silver-stained reducing 6% polyacrylamide gel electrophoresis of gill mucin produced a band at the origin with a smear entering the separating gel. There was no evidence of a link protein in gill mucin on reducing 12% polyacrylamide gel electrophoresis. Gill mucin had an amino acid profile similar to that of mucins in other species. Specifically, 35.1% of the total amino acids were represented by threonine and serine, while another 27.5% were alanine and proline. Gill mucin contained galactose (26.7 ± 3.2%), galactosamine (22.5 ± 4.4%), glucose (16.6 ± 8.7%), fucose (16.1 ± 1.5%), glucosamine (12.0 ± 1.9%) and mannose (5.1 ± 4.4%). Uronic acid levels from purified mucin were very low (0.7 ± 0.1%). Sialic acid was also present (0.06 g/g of mucin protein). The periodic acid-Schiff assay routinely utilized for detection of mucins was relatively insensitive for detection of gill mucin (6 × less sensitive than for pig gastric mucin) so a rabbit antiserum was raised. The antiserum produced profiles similar to the periodic acid-Schiff assay of fractions following gel filtration. Immunofluorescence of formalin-fixed rainbow trout gill tissue sections showed that the antiserum detected mucin within branchial goblet cells.     

Low feed pellet water stability and fluctuating water salinity cause separation and accumulation of dietary oil in the stomach of rainbow trout (Oncorhynchus mykiss)

Regurgitation (belching) of oil is a problem in sea farmed rainbow trout kept in water with fluctuating salinity. This often coincides with abdominal distension syndrome, which is associated with osmoregulatory stress and lipid indigestion. The present study investigated combined effects of water stability in extruded feeds and fluctuating water salinity on feed consumption, growth, nutrient digestibility, gastrointestinal transit, and accumulation of water and oil in the stomach of rainbow trout. Diets were produced with high or low water stability, which were characterised by a novel water stability test. In an initial 84 days growth experiment, each diet was fed to six groups of 772 g trout, of which three tanks were supplied with full-strength seawater, and three tanks were fluctuated between seawater and freshwater (7 h per day during 3 successive days per week). No significant effects were seen, although the feed intake appeared slightly lowered by low dietary water stability at fluctuating salinity. In a second experiment, each diet was fed to four groups of 1 kg trout, of which two tanks were supplied with full-strength seawater, and two tanks were fluctuated between seawater and brackish water (17‰ 7 h per day daily). Low water stability of the diet resulted in accumulation of oil in the stomach, and this response was worsened when the salinity of the rearing water fluctuated. It also corresponded with reduced lipid digestibility. Consequently, low water stability of the diet caused an oil separation in the stomach that may result in oil-belching in rainbow trout suffering from osmoregulatory stress.     

一株虹鳟源传染性胰腺坏死病病毒的分离与鉴定

为明确引起四川石棉某养殖场饲养的虹鳟患病死亡的病原体,实验对自然发病虹鳟进行大体病变观察并对其病原体进行分离,通过人工感染实验及多重RT-PCR鉴定确定病原体WZ160509,并对病原体的主要结构蛋白VP2进行扩增分析,同时对病变组织进行组织病理学观察。结果显示,患病鱼主要临床症状表现为体表发黑,腹部膨大,挤压腹部可见肛门喷射淡黄色黏液便;剖检可见肝脏、肾脏苍白;肠道内无食物,内积黄色黏液。将患病虹鳟组织匀浆液无菌接种虹鳟鱼生殖腺细胞系(rainbow trout gonad cell line,RTG-2)细胞,盲传3代均出现典型的细胞病变。人工感染实验显示死亡率高达90%,并出现与自然患病鱼相同的症状。多重RT-PCR检测发现,自然发病鱼、人工感染鱼以及病变RTG-2细胞均为传染性胰腺坏死病病毒(infectious pancreatic necrosis virus,IPNV)阳性,其主要结构蛋白VP2基因与美国分离株基因组1型聚为一支,且同源性分析表明,WZ160509-VP2与IPNV-VP2(AY026345)的同源性最高,序列一致性为95.8%。组织病理学观察显示,患病鱼胰腺细胞空泡变性,坏死;肝细胞空泡变性,坏死;肾小球轻度炎症,毛细血管通透性增加,肾小囊腔内有红色絮状蛋白类物质渗出,肾小管上皮细胞空泡变性。研究表明,从该养殖场患病虹鳟中分离到的病毒为IPNV。     

Insulin-receptor binding in skeletal muscle of trout

Two hundred rainbow trout (Oncorhynchus mykiss) age 0 +, weight range 11.3 – 11.5 g, were distributed randomly in two groups and maintained for five weeks on either 10% dextrin, or 20% dextrin diet. The fish were sampled 3–5 h and 18–20 h after the last feeding and insulin binding to partially purified insulin receptors in white and red skeletal muscles and to liver plasma membranes was assessed. Plasma insulin, plasma glucose, and liver glycogen content were analyzed in the same fish.Fish fed a diet with higher carbohydrate content (HC) had elevated insulin and glucose levels in peripheral blood, but lower liver glycogen contents compared to the fish fed a diet with lower carbohydrate content (LC). No growth retardation was observed in the fish from HC group.Three to five hours after the last feeding, insulin-receptor binding in white skeletal muscles was higher in HC group of fish, mostly because of an increase in number of high affinity binding sites. Eighteen to twenty hours after the last feeding this difference disappeared. In contrast, the specific binding of insulin to the liver plasma membranes appeared to be lower in the HC group of fish. The lower insulin binding to the liver plasma membranes observed 3–5 h after feeding, could be attributed to the lower quantity of binding sites, while the same phenomenon 18 h after feeding was likely a result of affinity changes. We conclude that higher glycemic levels observed in trout fed a HC diet as compared to LC group of fish, are not a consequence of impaired binding of insulin to its receptors in skeletal muscles.Presented in part at the Annual Meeting of American Society of Zoologists, December 26–30, 1989, Boston, U.S.A. Supported by a grant (Personal investigador en el extranjero) of Ministerio de Educacion y Ciencia, (CYCIT) Spain to J.G. and by grants of the U.S. National Science Foundation (DCB 8615551 and DCB 8915935 to E.P.) and North Atlantic Treaty Organization (NATO) 0926/87 to C. Ottolenghi and E.P.     

大西洋鲑、三倍体虹鳟和金鳟的肌肉营养成分与品质特性

为了解大西洋鲑(Salmo salar)、三倍体虹鳟(Oncorhynchus mykiss)、金鳟(Oncorhynchus mykiss)3种鱼肌肉营养成分和品质特性,利用生化分析、物性分析方法分析3种鱼肌肉的营养成分、氨基酸和脂肪酸组成、肉色、系水力和质构特性。结果表明,大西洋鲑、三倍体虹鳟、金鳟肌肉的水分质量分数分别为62.91%、67.15%、73.02%,粗蛋白质量分数分别为22.39%、21.03%、22.11%,粗脂肪质量分数分别为14.64%、17.16%、5.11%。3种鱼肌肉的滴水损失、黄色值(b~*)、羟脯氨酸含量、内聚性均显著不差异(P0.05)。3种鱼肌肉的硬度和咀嚼性由低到高依次为大西洋鲑、三倍体虹鳟、金鳟,而pH值的结果则与之相反(P0.05)。大西洋鲑和三倍体虹鳟肌肉的灰分、解冻损失、蒸煮损失、回复性、弹性和红色值(a~*)差异不显著(P0.05),但灰分、蒸煮损失、回复性均小于金鳟肌肉的对应指标(P0.05),弹性和红色值(a~*)则均大于金鳟肌肉的对应指标(P0.05)。大西洋鲑、三倍体虹鳟、金鳟肌肉中必需氨基酸含量占氨基酸总量分别为42.28%、41.84%、41.63%(质量分数),必需氨基酸/非必需氨基酸(EAA/NEAA)比值分别为73.25%、71.94%、71.32%,均符合联合国粮农组织/世界卫生组织(FAO/WHO)对优质蛋白质的评价标准;3种鱼肌肉中均检测到22种脂肪酸,组成丰富,其中不饱和脂肪酸含量较高。综上所述,3种鱼的肌肉都是符合人体营养需求的优质水产品,其中大西洋鲑和三倍体虹鳟肉质接近,且都优于金鳟的肉质。     

Assessment of “discreet” vertebral abnormalities, bone mineralization and bone compactness in farmed rainbow trout

The present study aimed to evaluate the importance of “discreet” vertebral abnormalities in normally-shaped rainbow trout in relation to vertebral bone condition in French fish farms. A total of 373 trout (262 ± 2 mm in total length) from 20 fish farms sampled were studied. The fish were radiographed and the axial skeleton examined for vertebral abnormalities. Vertebrae from the middle axial region (V32-38) were selected to evaluate vertebral bone condition. Bone mineralization (BM, %) was estimated by the ratio of ash and dry weight. Bone compactness (BC,%) was measured using Bone Profiler 3.23 images software on digitized radiographs of transverse sections (125 ± 10 μm). Statistical analyses were performed to test the relationships between the occurrence of vertebral abnormalities, and BM and BC. The occurrence of affected trout ranged from 0 to 55% depending on the farm. Trout displayed vertebrae with low BM (< 54.6%) and low BC (< 28.1%) in 40% and 55% of the farm, respectively. No relationships were observed between bone condition parameters (BM, BC) and the occurrence of vertebral abnormalities. These results could be explained by a wide and variable plastic response of bone characters (i.e., vertebral abnormalities, BM and BC) to the various rearing conditions in the fish farms sampled.     

虾青素和角黄素对虹鳟肌肉着色和肝脏总抗氧化能力的影响

分别在基础饲料(对照组)中添加100 mg/kg的虾青素、角黄素,混合色素(50 mg/kg虾青素+50 mg/kg角黄素)饲喂初始体重为(56.60±0.63) g的虹鳟60 d,考察虾青素和角黄素对虹鳟肌肉着色和肝脏总抗氧化能力的影响。结果显示,饲料中添加了虾青素、角黄素和混合色素后对虹鳟增重率、饲料系数及肌肉常规成分、肌肉失水率、含肉率均无显著影响(P> 0.05)。虾青素组、角黄素组和混合色素组虹鳟肌肉的比色卡得分、红度、虾青素含量和血清总类胡萝卜素含量均比对照组有显著提高(P< 0.05);虾青素组虹鳟肌肉比色卡得分(26.25)和红度值(18.40)显著高于角黄素组(22.38, 14.13)和混合色素组(24.00, 15.70)(P< 0.05);虾青素组虹鳟肌肉虾青素含量为4.75 mg/kg (30 d)和6.45 mg/kg (60 d),均显著高于混合色素组的3.87 mg/kg (30 d)和5.48 mg/kg (60 d)(P< 0.05);在虹鳟血清总类胡萝卜素含量方面,虾青素组 > 混合色素组 > 角黄素组;虾青素组、角黄素组、混合色素组虹鳟肝脏的总抗氧化能力之间无显著差异(P> 0.05),分别为2.39 U/mg,2.25 U/mg,2.39 U/mg,均较对照组(2.03 U/mg)显著提高(P< 0.05)。上述结果表明：饲料中添加100 mg/kg虾青素、角黄素及虾青素+角黄素混合(1∶1)均能有效改善虹鳟肌肉颜色,提高肝脏总抗氧化能力,虾青素、虾青素+角黄素混合(1∶1)对虹鳟肌肉的着色效果优于角黄素。     

虹鳟源嗜冷黄杆菌的分离鉴定及致病性研究

为确定患病虹鳟的病原,本实验从患病鱼溃烂肌肉中分离到2株细菌,分别命名为CH06和CH07,经回归感染证实分离菌株为导致此次虹鳟患病的病原菌,并进一步对其形态特征、理化特性、分子特征、血清型及耐药性进行分析。结果显示,CH06和CH07株在TYES琼脂平板上呈煎蛋状外观,产黄色素,氧化酶和过氧化氢酶呈阳性,能水解明胶和酪蛋白,不能水解淀粉,不能利用果糖、半乳糖和七叶苷等。16S rRNA比对结果显示,CH06和CH07株与嗜冷黄杆菌模式株NBRC 15942的同源性分别为99.35%和99.42%。综合菌株理化和分子特性确定CH06和CH07株为嗜冷黄杆菌。利用多重PCR方法鉴定CH06和CH07株的血清型均为1型(Fd型);多位点序列分型(MLST)分析表明,CH06和CH07株的基因型分别为ST-12和ST-78型,且均属于CC-ST10克隆型。人工感染结果显示,CH06和CH07株对虹鳟幼鱼具有较高致病性,其半致死浓度(LD₅₀)分别为7.1×10⁵和1.1×10⁵ CFU/mL,攻毒剂量与临床病症出现时间呈反比,从人工感染实验鱼的肌肉、脾脏等组织中可重新分离到嗜冷黄杆菌。组织病理变化显示,病鱼肝细胞肿胀,空泡变性,部分肝细胞溶解坏死,细胞核溶解消失;脾脏充血、出血,淋巴细胞减少,红细胞和含铁血黄素增多;肌纤维间隙增宽、断裂、弯曲不齐,部分肌细胞肌浆溶解呈蜂窝状。CH06和CH07株对10种抗菌药物的耐药谱略有不同,均对氨苄西林和甲氧苄啶-磺胺甲噁唑敏感;CH06株对恩诺沙星、氟苯尼考等耐药,而CH07株对恩诺沙星和氟苯尼考中度敏感。本研究首次报道了我国虹鳟源嗜冷黄杆菌的分离鉴定及生物学特性,以期为虹鳟细菌性冷水病的防控提供科学依据。     

国内虹鳟代表性养殖群体的高通量SNP芯片检测及遗传分析

本研究旨在对国内虹鳟(Oncorhynchus mykiss)代表性养殖群体开展全基因组水平的遗传评估。利用57K单核苷酸多态性(single nucleotide polymorphism,SNP)芯片,检测了来自不同地域的6个虹鳟养殖群体样本共计48尾,包括黑龙江虹鳟、黑龙江金鳟、四川虹鳟、四川金鳟、北京虹鳟和北京金鳟,共获得有效SNP位点50201个,在中国虹鳟中的多态比例达到97.7%,表明该芯片虽然基于美国和挪威虹鳟群体设计,但对中国群体同样具有良好的适用性。各群体最小等位基因频率均值为0.240~0.267,与国外主流养殖群体相近,黑龙江虹鳟、四川虹鳟和北京虹鳟群体内遗传多样性丰富,多态位点比例为83.6%~84.9%,与国外主流养殖群体相近,而黑龙江金鳟、四川金鳟和北京金鳟,多态位点比例相对较低,在60.2%~76.9%范围内。应用6个中国虹鳟群体和2个美国虹鳟群体数据开展系统发育分析、主成分分析和群体遗传结构STRUCTURE分析,结果显示8个群体可分为3个祖源类群,其中3个金鳟群体为遗传联系较紧密的一个类群,黑龙江虹鳟和北京虹鳟为一个类群,而四川虹鳟与2个美国虹鳟群体为一个类群,部分中国养殖群体中有显著离群个体存在,表明群体遗传背景不均一。本研究表明,高密度SNP芯片在我国虹鳟养殖群体遗传分析中具有广泛的应用前景,能够为种质资源评估、本土化良种培育、制种和引种工作提供基因组水平的参考信息。     

Pseudomonas putida infection in rainbow trout

A disease outbreak occurred during the Spring of 2006 among rainbow trout, Oncorhynchus mykiss, kept in growout tanks. During the outbreak 35% of the rainbow trout died. Typical clinical signs were ulceration on the dorsal surface of the fish. Based on biochemical characteristics and sequence of 16S rRNA, the causative bacteria isolated from diseased fish were identified as Pseudomonas putida. Histologically, skin ulcer initially was characterized by sloughed off epithelia and epithelial necrosis. In the advanced form of the disease, almost all layers of the skin down to the epidermis were lost. Outside of Japan, this study reports the first P. putida infection in fish.     

虹鳟生长性状的随机回归分析

生长和抗逆是水产动物遗传育种工作中最重要的农艺性状,虹鳟的生长性状关乎虹鳟规模化养殖的生产经济效益,为了从遗传上精细解析虹鳟的生长性状,我们从渤海、丹麦、挪威、唐纳森和加利福尼亚5个虹鳟(Oncorhynchus mykiss)种系间的双列杂交开始,进行了连续4代的继代选育。本研究测量了第4代总共4368个实验个体在516日龄、608日龄、668日龄、883日龄和1036日龄5个时间点的生长数据。采用随机回归测定日模型,对虹鳟生长性状进行了动态遗传分析。根据贝叶斯信息准则,确定3阶勒让德多项式为拟合体重和体长的加性遗传效应和永久环境效应变化的最优子模型。利用双变量随机回归模型同时分析体重和体长两个生长性状。它们的遗传力在400～1000日龄之间呈现递减趋势,分别从0.288下降到0.164和从0.469下降到0.186,并且在该生长区间内体长的遗传力始终高于体重的遗传力。无论体重还是体长性状,在不同日龄之间的遗传相关都随着生长间隔的增大而降低,但是两个性状在生长初期和后期之间的遗传相关较高(遗传相关系数0.75以上),尤其是体重(遗传相关系数0.85以上),该研究结果为虹鳟早期的遗传选育提供了理论支撑。两个性状之间在相同日龄之间的遗传相关均在0.75以上,在不同日龄之间的遗传相关随着生长间隔的增大由0.83下降到0.63。以上的研究结论为虹鳟生长性状(主要是体长和体重)的遗传选育提供了理论基础,同时也为虹鳟的体长和体重两个性状的联合选育提供了精确的遗传分析结果,由于两性状在前期有较高的遗传相关,因此建议在虹鳟生长前期(400日龄)进行联合选择。     

太平洋鳕RAG1和IgM基因荧光定量PCR方法的建立及初步应用

为研究太平洋鳕发育早期特异免疫系统形成的机制,通过RAG1和IgM基因的转录水平衡量特异免疫系统的发育特点.根据GenBank中RAG1和IgM的序列信息,分别设计1对特异引物,从太平洋鳕头肾中扩增得到RAG1和IgM的基因片段.将所获基因片段分别插入到克隆载体pMD18-T中,从而构建太平洋鳕RAG1和IgM基因的质粒标准品.建立并优化太平洋鳕RAG1和IgM基因绝对荧光定量PCR方法.为进一步验证该方法的可靠性,分别利用绝对定量和相对定量检验目的基因在太平洋鳕早期发育过程不同组织内的表达差异.以优化后的绝对荧光定量PCR方法检测不同发育时期太平洋鳕RAG1和IgM的表达情况.结果显示,RAG1的回归方程为y=-3.266x+33.77,回归系数R2=0.996;IgM的回归方程为y=-3.119x +27.61,回归系数R2 =0.998.绝对定量和相对定量结果在基因转录趋势上显现出一致性,即RAG1基因在胸腺和头肾中表达,且在胸腺中的表达量显著高于头肾中的表达量,在肝脏和脾脏中无表达;IgM基因在胸腺、头肾、肝脏和脾脏中均有表达,其中脾脏中表达量最高,其次是头肾.RAG1基因在太平洋鳕发育早期的表达水平很低,到61日龄(days posthatching,dph)至95 dph表达量显著提高;IgM基因在早期表达水平同样很低,到33 dph至61 dph才有明显表达,在95 dph时表达量显著提高.研究表明,本实验方法可靠,特异性较强,可成功对目标基因转录水平进行检测.     

AFLP-based genetic linkage map of marine shrimp Penaeus (Fenneropenaeus) chinensis

This paper presents the genetic linkage map of the Chinese shrimp Penaeus (Fenneropenaeus) chinensis constructed with 472 AFLP markers. A hundred F₁ progeny from an intercross between a female from the new variety “Yellow Sea No. 1” and wild caught male used for the mapping study. Two separate maps were constructed for each parent. The female linkage map consisted of 197 marker loci forming 35 linkage groups and spanned a total length of 2191.1 cM, with an average marker space of 13.5 cM. The male map consisted of 194 marker loci mapped to 36 linkage groups and spanned a total length of 1737.3 cM, with an average marker spacing of 11.0 cM. The level of segregation distortion observed in this study was 12.2%. The estimated genome length of P. chinensis was 3150.3 cM for the female and 2549.3 cM for the male, respectively. The observed genome coverage was 69.6% for the female and 68.1% for the male map. The linkage maps constructed in this study provide basic information for further linkage studies on Chinese shrimp, and more importantly, the construction of the maps are part of the work of the genetic breeding programs which will be used for growth discovered in the QTL analysis of P. chinensis.     

罗非鱼海豚链球菌病的病理学观察

采用光学显微镜和透射电子显微镜技术分别对患海豚链球菌病的罗非鱼的病理学变化进行了研究。分离的病原菌革兰氏染色呈阳性,透射电镜负染观察菌体球形或卵圆形,直径0.6~1.0μm,多数呈链状排列。组织学病变主要表现为全身多组织、器官水肿,出血、变性、坏死以及炎症反应,特别是肝、脾、肾和脑分别表现为肝炎,脾炎,间质性肾炎和脑膜炎。超微结构观察发现病鱼肝、脾、肾、脑、心肌和骨骼肌等器官的细胞超微结构都有较为严重的破坏,细胞核畸形,染色质浓缩或边集,粗面内质网囊泡化及脱粒,线粒体肿胀,嵴断裂或溶解消失。研究表明,海豚链球菌能造成罗非鱼全身性组织器官病变,致使器官功能障碍,正常生理代谢调节紊乱,最后导致死亡。     

Disease of rainbow trout caused by Pseudomonas luteola

Bacteria isolated from rainbow trout, Oncorhynchus mykiss, kept in a farm, in Turkey. During the outbreak, 40% of the rainbow trout (10-40 g) died. Typical clinical signs were exophthalmia, dark pigmentation, hemorrhage at the base of the pectoral, pelvic, anal fins and around the vent. Internal signs were enlarged spleen, pale liver and intestine filled with yellowish fluid. Liver, kidney and spleen of diseased fish, were aseptically streaked on Tryptic Soy Agar. After incubation, pure cultured colonies were observed and biochemically characterized with API 20 NE and other biochemical tests. Cultured bacterial 16 S rDNA gene was sequenced. Based on biochemical characteristics and sequence of 16 S rRNA, the causative bacteria were identified as Pseudomonas luteola. This study reports the first P. luteola infection in fish.