新型鸭呼肠孤病毒RT-PCR检测方法建立与初步应用

为建立一种能够快速检测新型鸭呼肠孤病毒（NDRV）的方法,本研究参考GenBank上登录的NDRV S3基因保守序列设计特异性引物.经条件优化后,建立了检测NDRV的RT-PCR方法.对其特异性、敏感性和重复性进行检验.结果显示：该方法仅能从NDRV分离毒中扩增到与预期大小相符,长度为298 bp的特异性目的片段,检测灵敏度达到83.4 pg病毒RNA;而其它病毒：番鸭呼肠孤病毒、禽呼肠孤病毒、鸭病毒性肝炎病毒、鸭瘟病毒、鸭新城疫病毒和禽流感病毒等样品的扩增结果均为阴性.采用该方法对在广东不同地区采集的15份鸭病料样品进行检测,NDRV阳性率为53.33％.表明建立的RT-PCR方法特异性强、敏感度高,可用于NDRV的临床诊断和流行病学调查.     

不同给药方式下培氟沙星在鲤体内的药代动力学研究

在水温为18℃下,按10 mg/kg（体质量）对体质量为（200±30）g的福瑞鲤Cyprinus carpio单次肌肉注射和混饲口灌培氟沙星,于不同时间点采集鲤血浆、肌肉、肝胰脏、肾脏组织,经超高液相色谱法测定各组织中培氟沙星的浓度,并采用DAS 3.0药物代谢动力学软件的非房室模型统计矩方法分析药时数据。结果表明：混饲口灌给药和肌注给药后,培氟沙星的药时曲线下面积（ AUC）分别为88.35、139.9 mg·h/L,达峰浓度（ Cmax ）分别为2.092、3.687 mg/L,达峰时间（ Tmax ）分别为4.0、0.5 h,消除半衰期（ t1/2）分别为22.301、74.357 h,表观分布容积（ Vd ）分别为5.464、15.342 L/kg,总体清除率（ CL ）分别为0.170、0.143 L·kg/h。研究表明,肌肉注射给药较混饲口灌达峰时间短,达峰浓度高,半衰期长,生物利用度高。     

几种淡水养殖鱼死后僵硬的季节变化

几种淡水养殖鱼活杀死后在室全僵的时间（Ｔｒ）同其生活时的水温（ｔ）之间存在着良好的负相关，即ｌｎＴｒ＝５．０３７－０．１１６５ｔ（ｒ＝－０．９２３６），显示出明显的季节差异。死后僵硬还受致死方法影响，表现为活杀鱼的僵硬时间长，挣扎死的短，半死的近于二者之间。据此，讨论了在温度低，僵硬期长的季节，采用冰藏或１０℃以常温条件下活杀鲜鱼流通上市，拓宽淡水养殖鱼类流通渠道的可能性及活杀鲜鱼的鲜度质量指标等     

草鱼出血病细胞培养弱毒疫苗的制备及其免疫效果

本文报道经筛选的ＧＣＨＶ－８４１毒株在草鱼吻端成纤维细胞系中进行适应性培养，弱毒驯化，细胞培养弱毒疫苗的制备，安全性观察，不同稀释度和剂量的免疫保护率测定及保存试验，ＧＣＨＶ－８４１毒株在ＰＳＦ细胞中继代驯化至５３代达到减毒目的。筛选得５３－５９代作毒株在ＰＳＦ细胞中继代驯化至５３代达到减毒目的，筛选得５３－５９代作毒种制备的细胞制备细胞弱毒疫使用安全，用稀释１０－１０＾４倍的疫苗，０．１－０．３     

超高压对草鱼肌肉超微结构与质构特性的影响

研究了等静超高压对草鱼肌肉超微结构与质构特性的影响，并与猪肌肉用对照。结果表明：１．加压处理的草鱼肌肉，其超微结构中出现较明显的变化，如肌原纤维中Ａ带和Ｉ带的细丝均被破坏，粗丝相互聚集，出现间隙，Ｚ线变粗，不连续，Ｈ带和Ｍ线消失等，但肌原纤维的外形仍保持完整。２加压处理的草鱼肌肉，其外观无显著变化，色泽稍白，略有汁液流出，但质构特性与处理前无明显差别。     

Growth testing of two breeds of common carp, Cyprinus carpio L. (Hungarian mirror and Třeboň scaly carp), in ponds with low and high stocking density

The growth of two breeds of common carp, Cyprinus carpio L., was tested in ponds under the climatic conditions of South Bohemia. T?eboň scaly carp (TR) and Hungarian mirror carp (M2) were kept in both low and high stocking densities during the second growing season and then stocked together for communal testing during the third growing season. Before the communal testing, the mean initial weights of fish from low‐ and high‐density stocks differed significantly (374.1 vs. 227.7 g for the TR breed and 766.7 vs. 317.3 g for the M2 breed respectively, P<0.01). After communal testing, mean weights of fish from low‐ and high‐density stocks gained 761.8 vs. 543.8 g for the TR breed and 1339.7 vs. 706.7 g for the M2 breed respectively. These observed weights were also significantly different (P<0.01). However, the test of corrected weight gain, i.e. gain not related to the initial weight of fish, revealed insignificant differences (P>0.01) between the weight gains after correction, i.e. the effect of different initial weights was successfully eliminated. These results seem to confirm the applicability of this method for the assessment of growth of purebred common carp under the climatic conditions of Central European fish farms.     

The evaluation of secondary stress response of grass carp (Ctenopharyngodon idella, Val. 1844) after exposing to the saline water

The stress response of grass carp to salinity was evaluated assessing the levels of hematocrit, leucocrit, plasma glucose, sodium, potassium and calcium. Mean hematocrit values increased following exposure to salinity. Mean leucocrit values elevated after exposure to salinity for 24 h, then decreased. Mean plasma glucose increased during saline exposure for 48 h and then decreased to normal values. Plasma sodium and chloride levels were increased to their highest values after 48 h. While plasma potassium value increased with the exposure time, plasma calcium values were not affected by salinity exposure.     

Dynamics of appetite in three fish species (Gasterosteus aculeatus, Phoxinus phoxinus and Carassius auratus gibelio) after feed deprivation

The temporal dynamics of appetite (weight of food consumed per day) were analysed for fish fed to satiation after a 1 or 2-week period of feed deprivation. Three species were compared: two omnivores (minnow, Phoxinus phoxinus and gibel carp, Carassius auratus gibelio) and a carnivore (three-spined stickleback, Gasterosteus aculeatus). All species showed compensatory changes in appetite and growth during the re-feeding period but in the stickleback, there was a lag of a week before the compensatory response was detected. The temporal dynamics of appetite differed between the three species but not within a species. Appetite of the minnow declined towards control levels from the onset of re-feeding. In gibel carp, appetite increased to a peak and then declined to control levels. In the stickleback, appetite was initially below control levels, increased to a maximum, then declined towards control levels. The differences between the species might have been artefacts of the experimental protocols used but could also reflect underlying differences in the control of appetite in these species of fish.     

The purification and partial characterization of carp,Cyprinus carpio,vitellogenin

A procedure is described for the isolation of intact vitellogenin (c-VTG) from the carp, Cyprinus carpio. VTG was induced in juvenile females using oestradiol-17β and purified from the plasma using a combination of gel-filtration chromatography on Sepharose 6B and ion exchange chromatography on DEAE-cellulose. Purification procedures were conducted at low temperatures (below 9°C) in the presence of the proteolytic enzyme inhibitor aprotinin to prevent degradation. Intact c-VTG had an apparent molecular mass of 390,000 Daltons, but when extracted from plasma in the absence of aprotinin it underwent proteolysis into at least 2 protein fragments (apparent molecular masses of 230,000 and 96,000 Daltons), showing an instability of the native dimer. An amino acid analysis of c-VTG showed that its composition was almost identical to goldfish VTG, a species closely allied to the true carps and also similar to other oviparous vertebrate VTGs. Collectively, these data indicate that using these purification procedures VTG from carp, and probably other teleost species, can be isolated in an intact, highly purified form.     

草鱼出血病的病原研究

草鱼出血病的病原研究,始于50年代。1978—84年,分离到一种病原病毒,定名为草鱼呼肠孤病毒或鱼呼肠孤病毒。本文报道从出血病病鱼组织的电镜研究中发现两种病毒颗料,一种即是呼肠孤病毒,另一种是20-30nm大小的病毒。经病毒的核酸分析,前者是双股RNA病毒;后者为单股RNA病毒。用分离到的这两种病毒分别注入1足龄健康草鱼,可发生两类不同症状的出血病;呼肠孤病毒主要导致“肠出血型”症状;另一种病毒(经初步鉴别属于小RNA病毒科病毒)主要导致“肌肉出血型”出血病。由此,可以证实两种病毒都是草鱼出血病的病原病毒,同时也初步解释出现两种不同症状出血病的原因。