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。     

草鱼肠道、肝胰脏对饲料蛋白质酶解速度的比较

草鱼样品体重(1543±213)g。采用离体消化培养和茚三酮比色方法,以草鱼前肠、中肠、后肠和肝胰脏的粗酶液作为酶源,对鱼粉、豆粕、菜粕和棉粕进行酶解,并测定酶解14h内,豆粕酶解液中氨基酸生成量随时间的变化,以及0至4h,内4种饲料蛋白质酶解液中氨基酸的生成量。结果表明,(1)在0至4h内,4种饲料酶解液OD570随时间变化的线性关系较好。(2)对于同种蛋白质饲料原料,以中肠组织粗酶液的酶解速度最大,其余依次为前肠、后肠及肝胰脏粗酶液。表明草鱼中肠对饲料蛋白质的酶解能力强于前肠和后肠,同时,肠道酶液的酶解速度均大于肝胰脏。(3)对于4种不同的蛋白质饲料原料,以鱼粉酶解氨基酸生成速度最大,其他依次为豆粕、菜粕、棉粕。(4)草鱼肝胰脏粗酶液对豆粕的酶解氨基酸生成速度最大,表明草鱼对豆粕进行酶解消化的能力较强。实验证实,饲料原料种类的差异和饲料蛋白质组成与性质的差异使其酶解氨基酸生成量和生成速度有差异。     

Effects of feeding rates on growth and production of Nile tilapia, common carp and silver carp polycultured in fertilized ponds

Abstract In an attempt to identify appropriate feeding rates for multispecies of fish raised in fertilized earthen ponds, the present work was conducted over a 19‐week experimental period to establish the growth performance, production and body composition of Nile tilapia, common carp and silver carp fed 0.0%, 0.5%, 1.0%, 3.0%, 5.0% biomass and to apparent satiation (treatments). Twelve ponds were stocked with a similar number and weight of each fish species. Two ponds were assigned to each of the treatments, and a 25% protein pelleted fish feed was used to feed fish at the specified rate of feeding. At the end of the experiment, growth, weight gain, survival, yield and body composition of fish groups were affected by the treatments. The economic effectiveness also varied among treatments. The most conspicuous attribute of the feeding rates was its lack of influence on growth (g day^?1), weight gain (g per fish), yield (kg ha^?1) or body composition of silver carp. The results of whole‐body proximate analysis indicated that various feeding rates had either an irregular pattern or no effects on the protein and ash gain per 100 g of fish body weight (bw) gain. The most notable exceptions were significant (P < 0.05) increases in body fat and gross energy gains in Nile tilapia, common carp and silver carp accompanied by decreases in percentages of moisture (but not in silver carp) as feeding rate increased. Among the six different feeding levels, feeding to apparent satiation (feed amount was equivalent to 2.67% of fish bw day^?1) appeared to be optimal, as it significantly (P < 0.05) supported the highest fish production, income and net profit compared with all other treatments except for the 3% feeding level, for which the differences in those measurements were comparable.     

Artificial spawning of carp (Cyprinus carpio L.); differences between the effects of reproduction in females of Hungarian, Polish and French origin treated with carp pituitary homogenate or [D-Tle6, ProNHEt9] GnRH (Lecirelin)

The investigation concerned the reproduction effects in carp females of the Hungarian strain 8, Polish strain 2 and French strain F of ovulation stimulation with carp pituitary homogenate (0.3 mg kg^?1 and after 12 h 2.7 mg kg^?1) or [D‐Tle⁶, ProNHEt⁹]GnRH (Lecirelin) with metoclopramide (15 μg kg^?1 and 10 mg kg^?1 respectively). The ovulation stimulators did not affect the weight of eggs (expressed in g and as the percentage of female body weight). However, in the group treated with pituitary homogenate (I) the least‐square means estimated for these parameters were higher than in the group treated with Lecirelin (II). A statistically significant (P≤0.05) difference in the mean percentage of living embryos after 48‐h incubation was observed between groups I and II, with the eggs obtained from females of group I being of better quality. The origin of females significantly (P≤0.05) affected the weight of eggs. The weight of eggs from females of Hungarian strain 8 was higher (P≤0.05) than the weight of eggs of the other two lines. With respect to parameters for egg quality (percentage of fertilized eggs and percentage of living embryos after 24‐, 36‐ and 48‐h incubation), no effect of the origin of females was observed. The interaction between the spawning agent and the origin of the females was not statistically significant with respect to the two parameters for the weight of the eggs. The least‐square means test for the investigated interaction showed that after the application of pituitary homogenate, the weight of eggs obtained from strains 8 and F was similar (respectively 711.2 g and 665.0 g). However, after the application of Lecirelin the females of strain 8 yielded eggs of a high weight (934.3 g) and those of strain F of a very low weight (373.2 g). A statistically significant (P≤0.05) interaction between ovulation stimulator and origin of females was recorded for the percentage of living embryos after 24‐ and 36‐h incubation. A dependence was found between the latency time and the reproduction effects.     

鲤鱼饲料添加剂的研究

根据鲤鱼的基本营养需求，确定基础饲料（蛋白质，脂肪，碳水化合物）配方后，对１２种无机盐和１２种维生素用数理统计正交试验，设计三个水平Ｌ２７（３＾１３），求出它们的电佳添加量，从而得出符合鲤鱼生理需求的无机盐，维生素综合配方－ＧＳ８９１添加剂。该添加剂经生产规模试验（池塘养鲤，水库网箱养鲤）证实，具有明显的促生长和节约饲料的功效。同期对比，增重率提高３０％以上，饲料系数降低７．９％以上。     

大头鲤粘孢子虫病的治疗试验

对患粘孢子虫病的大头鲤分别用食盐、亚甲基蓝、硝酸亚汞、敌百虫、硫酸亚铁和孔雀石绿几种药物进行治疗。结果表明 ,敌百虫与硫酸亚铁合剂以 1mg·L- 1 :0 2mg·L- 1 溶液治疗效果最好 ,治愈率 80 % ;1mg·L- 1 敌百虫溶液的效果次之 ,治愈率 5 0 % ;用0 1mg·L- 1 硝酸亚汞溶液的治愈率为 1 0 % ;用食盐溶液、亚甲基蓝溶液与孔雀石绿溶液处理无明显治疗效果。     

巴氏杀菌加热中3种淡水鱼肌肉的变性研究

将鲤、鲢和罗非鱼肌肉在巴氏杀菌条件 ( 75℃和 80℃ )下加热 ,测定其Vp值、升温曲线的斜率 fh 值、失水率以及过氧化物酶活力的变化特点。结果表明 ,3种鱼中罗非鱼肌肉升温速度最快 ,鲤最慢 ;罗非鱼肌肉过氧化物酶完全失活所需的Vp值最高 ;随加热温度升高 ,鲢失水率增加最多 ,而罗非鱼变化最小。在巴氏杀菌条件下 ,不同鱼类肌肉失水率对加热温度、Vp值变化的敏感性存在较大差异。因此 ,在生产中应根据加热温度、Vp、失水率等参数合理制定各鱼种相应的巴氏杀菌工艺条件     

青鱼消化器官数量性状胚后发育规律

采用显微解剖和测量、光镜和扫描电镜观察方法，系统观测全长为５．６～３３０．０ｍｍ的青鱼标本３７０尾，描述了鳃耙、咽齿、角质垫和肠等消化器官数量性状在胚后发育过程中的变化规律；论述了青鱼的摄食特性、消化器官数量性状胚后发育的阶段性及其相关性、消化器官数量性状发育与食性转化，为制订培育青鱼苗种和饲养成鱼的生物学技术提供了可靠依据     

天然雌核发育普安鲫(A型)细胞遗传学和血清电泳的初步研究

本文对贵州省普安县青山一带天然水体中的普安鲫(A型)作了细胞遗传学观察和血清电泳分析,发现它是天然雌核发育的鲫鱼,既不同于普通野鲫,也与已知的天然雌核发育方正银鲫和滇池高背鲫不是同一来源。     

Interaction of Grazing Muzzle Use and Grass Species on Forage Intake of Horses

The impact of horse preference and grass morphology on grazing muzzle effectiveness has not been investigated. The objective of this study was to determine the effectiveness of grazing muzzle use at reducing forage intake when horses grazed grasses with different morphology and preferences. The study was conducted in 2012 and 2013. Four horses were grazed in 2012, and three horses were grazed in 2013. Four species of perennial cool-season grasses were grazed in 2012 including Kentucky bluegrass (Poa pratensis L.), meadow fescue (Schedonorus pratensis Huds.), perennial ryegrass (Lolium perenne L.), and reed canarygrass (Phalaris arundinacea L.). In 2013, only Kentucky bluegrass and reed canarygrass were grazed because of winter kill of other species. Horses were allowed to graze a small pasture seeded with an individual species for 4 hours each day in June and August of 2012 and August and September of 2013. Horses grazed the same grass species for two consecutive days, one day with a muzzle and one day without. Before and after each grazing, a strip was mechanically harvested to determine initial and residual herbage mass. The difference was used to estimate forage intake. The effectiveness of a grazing muzzle was not affected by forage species (P ≥ .05). Use of a grazing muzzle decreased the amount of forage consumed by an average of 30% compared with not using a grazing muzzle (P < .0001). Results will aid horse owners and professionals in estimating forage intake of muzzled horses on pasture.