草鱼巨噬细胞的集结及相关的组织变化

给草鱼腹腔注射细微炭粒（墨汁）后不同时间取样，比较观察了草鱼胸腺、肝、脾、头肾和肾组织中巨噬细胞集结（ｍ ａｃｒｏｐｈａｇｅ ａｇｇｒｅｇａｔｉｏｎ ， Ｍ Ａ）状况及相应组织的变化。结果表明，在头肾和脾组织中， Ｍ Ａ 的数量和体积均较肝和肾中的多而大，胸腺中所产生的 Ｍ Ａ 数量较肝和肾中的少，体积也小；注射墨汁后 １～３ 周内，在胸腺、头肾、肝和脾组织中， Ｍ Ａ 的数量和体积均有不同程度的增加，但在注射墨汁后第 ４ 周的肾组织中， Ｍ Ａ 的体积缩小而且密度降低，同时肾组织结构有逐渐恢复的趋势，可能是由于 Ｍ Ａ 分解后通过肾小球和肾小管排泄到膀胱或体外的缘故。对草鱼 Ｍ Ａ 的组织分布、排除等分析认为， Ｍ Ａ 可作为草鱼非特异细胞防御作用的解剖学和细胞学的一种生物标记。     

利用紫苏子提取物作为异育银鲫饲料添加剂的生产试验

通过在异育银鲫饲料中添加300mg/kg紫苏子提取物,经随机抽取两处10口池塘,计算同期池塘应用及未应用紫苏子提取物添加饲料池塘投入产出情况:应用紫苏子提取物作为添加剂的池塘,异育银鲫增重率比未使用紫苏子提取物的池塘高9%～15%;饲料系数下降11.4%～11.8%,异育银鲫重量增长了50～99kg/亩,产值增长680～853元/亩。     

Essential oil of Aloysia citriodora Paláu and citral: sedative and anesthetic efficacy and safety in Rhamdia quelen and Ctenopharyngodon idella

ObjectiveTo verify the efficacy of citral in inducing sedation and anesthesia in silver catfish (Rhamdia quelen) and grass carp (Ctenopharyngodon idella) and to assess the safety of essential oil (EO) of Aloysia citriodora and citral in inducing and maintaining anesthesia in silver catfish.Study designClinical study, randomized, parallel, multi-arm with control group in target species.AnimalsA total of 96 juvenile and 72 adult silver catfish and 80 juvenile grass carp were used.MethodsSilver catfish and grass carp were exposed to different concentrations of citral, 15–675 and 15–600 μL L^–1, respectively, during the maximum period of 30 minutes to verify sedation and anesthesia induction and recovery times. In addition, for anesthetic induction, silver catfish were exposed to the EO of A. citriodora and citral at 225 μL L^–1 for 3.5 minutes. Then, fish were transferred to an anesthesia maintenance solution at 50 μL L^–1 for 10 minutes to assess hematologic and biochemical variables at 60 minutes, 2 and 6 days after treatment.ResultsCitral only induced sedation from 15, 25 and 40 μLL^–1 in both species. Anesthesia without mortality was induced in silver catfish at 50–600 μL L^–1 and grass carp at 75–450 μL L^–1. At 675 and 600 μL L^–1, mortality was recorded in silver catfish and grass carp, respectively. The EO of A. citriodora and citral were safe in inducing and maintaining anesthesia in silver catfish, with mean corpuscular hemoglobin concentration being the only variable that varied in relation to time and treatments.Conclusions and clinical relevanceCitral was effective in inducing sedation and anesthesia in both species. In addition, A. citriodora EO and citral were safe in inducing and maintaining anesthesia in silver catfish. Both agents are promising substances for the development of new drugs for fish.     

纳米氧化物对美托洛尔生物富集的影响研究

为揭示氧化物纳米颗粒（NPs）对离子型有机污染物在水生生物体内富集的影响,本研究考察了两种典型的氧化物NPs对鲤鱼富集β-阻断剂美托洛尔的影响以及美托洛尔在鱼体内各部位的分布,并通过批平衡实验进一步探讨了吸附解吸与生物富集的关系。结果表明：NPs共暴露时,在摄取阶段结束时美托洛尔生物浓缩因子（BCF）值增加了2.39倍（TiO₂ NPs）和3.49倍（SiO₂ NPs）。净化阶段鱼体内的美托洛尔半衰期由20.09d缩短到8.39d（TiO₂ NPs）和6.13d（SiO₂ NPs）,净化结束时鱼体内的美托洛尔残余浓度则略有升高。NPs的共暴露未改变鲤鱼摄取美托洛尔的途径,但显著升高了鱼鳃和内脏中美托洛尔的浓度。相对于TiO₂ NPs,SiO₂ NPs对美托洛尔具有更强的、且不可逆的吸附,使用Freundlich模型拟合的lg K_F值分别为2.21（TiO₂ NPs）和4.47（SiO₂ NPs）。研究表明NPs能够通过吸附载带促进鱼体对美托洛尔的富集,吸附态美托洛尔主要通过摄食和鳃的呼吸作用随NPs进入鱼体内,由NPs携带进入鱼体内的美托洛尔一部分发生解吸从而被鱼体利用,未发生解吸的部分随NPs的排出被排出体外。     

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。