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。     

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。     

诺氟沙星在金鳟体内的药物代谢动力学研究

实验水温为15±2℃,金鳟（Oncorhynchus mykiss）（平均体质量100±10g）单剂量肌肉注射30.0 mg/kg诺氟沙星后,应用高效液相色谱（HPCL）法于0.15,0.25,0.5,0.75,1,1.5,2,4,6,8,12,24,48,72 h测定了鱼血浆、肝脏和肾脏组织中药物的浓度,研究了诺氟沙星在金鳟组织中的分布及药物动力学规律。结果表明,诺氟沙星在金鳟体内吸收分布迅速,符合药物动力学一级吸收二室开放模型,但消除缓慢。诺氟沙星在金鳟血浆、肝脏和肾脏中的主要动力学参数如下：分布半衰期（T1/2α）分别为0.866、1.985、0.388h;消除半衰期（T1/2β）分别为31.369、36.402、30.975h;药时曲线下面积（AUC）分别为：308.005μg/mL.h、622.721μg/g.h、794.362μg/g.h。     

诺氟沙星在金鳟体内的药物代谢动力学研究

实验水温为15±2℃,金鳟（Oncorhynchus mykiss）（平均体质量100±10g）单剂量肌肉注射30.0 mg/kg诺氟沙星后,应用高效液相色谱（HPCL）法于0.15,0.25,0.5,0.75,1,1.5,2,4,6,8,12,24,48,72 h测定了鱼血浆、肝脏和肾脏组织中药物的浓度,研究了诺氟沙...     

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)。     

Production Characteristics of Rainbow Trout,Oncorhynchus mykiss and Brook Trout,Salvelinus fontinalis Under Seasonal Pond Conditions

Abstract

The production characteristics of juvenile rainbow trout, Oncorhynchus mykiss and brook trout, Salvelinus fontinalis were compared under winter pond conditions. Juvenile rainbow trout (55.1 ±1.5 g) and brook trout (28.9 ±0.4 g) were stocked at a density of 8,750 fish/ha into six 0.04-ha ponds. After 163 days, survival, growth, and feed conversion were similar (P >0.05). The results of this study suggest that brook trout may attain growth rates similar to rainbow trout under winter pond conditions in temperate regions of North America.     

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.     

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。     

Effect of GnRHa injection on milt volume in recently stripped rainbow trout Oncorhynchus mykiss

In this study, the effect of gonadotropin‐releasing hormone agonist (GnRHa) injection on milt production in spent rainbow trout was investigated. On day 0, 25 newly matured male rainbow trout (Oncorhynchus mykiss) were stripped manually, and sperm quantity (vol: mL fish^?1) and quality, spermatocrit (%), sperm count (cell mL^?1), motile sperm percentage and motility duration (s) were evaluated. After stripping, fish were randomly divided into five groups: intact; sham (injected with propylene glycol as a hormone vehicle); and groups receiving 4, 8 or 16 μg kg^?1 BW of [d ‐Ala⁶ Des‐Gly¹⁰] mGnRHa. On day 7, the fish were stripped again and the same sperm characteristics as on day 0 were measured. At the beginning of the experiment, there were no significant differences in any of the sperm quantity characteristics between groups. On day 7, expressible milt volume was significantly reduced compared with day 0 (P<0.05, t‐test) in the intact and sham groups but milt quality remained the same (P>0.05, t‐test). The present study shows that GnRHa injection with a concentration as low as 4 μg kg^?1 BW after first stripping could prevent a significant reduction in milt quantity collected 7 days later without any adverse effects on sperm quality.     

Effects of Microbial Phytase Supplementation in Corn Distiller's Dried Grain with Solubles on Nutrient Digestibility and Growth Performance of Rainbow Trout,Oncorhynchus mykiss

Abstract

Two experiments were conducted to evaluate the effects of microbial phytase in corn distiller's dried grain with solubles (DDGS) on apparent digestibility coefficients (ADCs) and growth performance of rainbow trout, Oncorhynchus mykiss. In experiment 1, DDGS was supplemented with phytase at 0,300,600,900, and 1200 FTU/kg diet. A total of 180 fish (129.1±9.3 g) were stocked into six 140-L tanks with 30 fish/tank. Fish were assigned randomly to a reference diet and each of the 5 diets containing DDGS. Yttrium oxide was used as an inert marker. Results showed that ADCs in DDGS supplemented with different dosages of phytase were: dry matter, 49.1-58.6%; crude fat, 78.9-88.9%; crude protein, 80.0-91.9%; gross energy, 50.5-66.6%; minerals, -7.3-99.7%; and amino acids: 73.9 to 96.8%. In experiment 2, abasal diet containing 15% DDGS supplemented with lysine and methionine was used to determine if trace mineral supplemental levels in rainbow trout diets could be reduced if microbial phytase was supplemented. Six diets were made with trace mineral premix supplementation at 0.1, 0.08, 0.06, 0.04, 0.02, and 0%. Phytase was not supplemented in the basal diet, but supplemented at 500 FTU/kg diet in all other diets. Ten-week results showed that there were no significant differences in fish weight gain, feed conversion ratio, and survival (P < 0.05). There were no significant differences in fish body composition and apparent nutrient retention among fish fed all diets, except that fish fed a diet without trace mineral supplementation had the lowest zinc level and the highest manganese retention. Results indicated that phytase was effective in releasing most of minerals, and that trace mineral supplementation level could be reduced when phytase was used in rainbow trout diets.     

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。     

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。     

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。     

The Economic Impact of the Trout,Oncorhynchus mykiss,Industry on Transylvania County,North Carolina

Abstract

The commercial culture of rainbow trout, Oncorhynchus mykiss, in Transylvania County, North Carolina, generated approximately $9 million in economic output in 2002. Commercial trout farming began in North Carolina more than 45 years ago. Twenty of the 57 commercial trout farms in production in North Carolina are located in Transylvania County, ranging in size from 1,135 to 204,300 kg annual production in 2002. North Carolina presently ranks second in the United States in commercial trout production, behind Idaho. The study shows that, in 2002, the trout industry generated about $9 million in economic output, created 201 jobs, and generated $3 and $0.9 million, respectively, as labor income and tax revenue in 2002. This is particularly important for Transylvania County where economic prosperity depends on locally available jobs and diversification of economic activities.     

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.     

The effectiveness of adenoviral vectors to deliver and express genes in rainbow trout,Oncorhynchus mykiss (Walbaum)

The efficacy of adenoviral vectors for gene delivery into fish cells, both in vitro and in vivo, was evaluated. Vectors utilized were of human adenovirus serotype 5 (Ad), which are commonly used in human clinical trials, but have not been assessed for gene delivery to fish. Because nothing is known about Ad receptors in fish, both an Ad (Ad5Luc1) with natural tropism for the coxsackie and adenovirus receptor (CAR), as well as an infectivity enhanced Ad (Ad5LucRGD) were included within this study. Gene expression was detected in cell lines using either vector. The levels seen with Ad5LucRGD were much higher than for Ad5Luc1 in most lines except CHSE-214. Transduction of CHSE-214 cells with Ad5Luc1 could be blocked with an excess of a competitive inhibitor, suggesting that these cells possess a CAR homologue thatmediates attachment of Ad, similar to that seen in mammalian cells. In vivo gene delivery was attempted by several methods, with significant expression seen only via intramuscular injection, although infection efficiency was low. Thus it was observed that several teleost cell lines are capable of being infected and one cell line expressed a human serotype adenoviral receptor homologue that aids in Ad infection. Additionally, in vivo studies indicated that muscle tissue of rainbow trout could be infected with Ad vectors, suggesting an alternative gene delivery strategy for this animal.     

三倍体虹鳟实时荧光定量PCR内参基因稳定性分析

采用实时荧光定量PCR(qRT-PCR)方法比较甘油醛-3-磷酸脱氢酶(gapdh)、核糖体大亚基蛋白基因(rplp2)、真核延伸因子基因(ef1-α)、β-肌动蛋白(β-actin)和18S核糖体核酸基因(18S rRNA)等5个候选内参基因在体质量约800 g三倍体虹鳟脑、眼、鳃、皮肤、心脏、肾脏(头肾、中肾、后肾...     

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.     

Utilization of (3S,3'S)-astaxanthin acyl esters in pigmentation of rainbow trout (Oncorhynchus mykiss)

The deposition of natural, optically active, astaxanthin fatty acid esters in rainbow trout ( Oncorhynchus mykiss ) was studied. Mono-esterified and di-esterified (3 S ,3' S ) astaxanthin were purified from the green microalga Haematococcus pluvialis and incorporated into extruded diets and compared with diets containing synthetic racemic astaxanthin (Carophyll Pink) and a total carotenoid extract from the alga. All sources of astaxanthin achieved >4 mg kg⁻¹ in the white muscle after 6 weeks feeding. No significant difference ( P  > 0.05) between the deposition of astaxanthin or total carotenoid for the different diets was observed. Other xanthophylls, namely lutein, zeaxanthin and idoxanthin were found in the white muscle of rainbow trout fed all diets and together accounted for 10–14% of total carotenoid. Astaxanthin was deposited in the white muscle in the stereochemical form administered in the diet, i.e. racemic astaxanthin for Carophyll Pink and ˜100% (3 S ,3' S )-astaxanthin for the algal sources. In contrast, epimerization of (3 S ,3' S ) astaxanthin from the alga was observed for the astaxanthin esters deposited in the skin of rainbow trout, with a ratio close to 1.0:2.0:1.0 (3 S ,3' S :3 R ,3' S :3 R ,3' R ).