Generation and characterization of aptamers against grass carp reovirus infection for the development of rapid detection assay     

Qing Yu  Mingzhu Liu  Siting Wu  Hehe Xiao  Xinling Qin  Pengfei Li 《Journal of fish diseases》2021,44(1):33-44

Grass carp reovirus (GCRV) causes devastating viral haemorrhagic disease in farmed grass carp (Ctenopharyngon idellus). As novel molecular probes, aptamers have been widely applied in rapid diagnosis and efficient therapies against virus or diseases. In this study, three single‐stranded DNA (ssDNA) aptamers were selected against GCRV‐infected CIK cells via SELEX (systematic evolution of ligands by exponential enrichment technology). Secondary structures predicted by MFOLD indicated that aptamers formed stem‐loop structures, and GVI‐11 had the lowest ΔG value of ?30.84 KJ/mol. Three aptamers could specifically recognize GCRV‐infected CIK cells, with calculated dissociation constants (Kd) of 220.86, 176.63 and 278.66 nM for aptamers GVI‐1, GVI‐7 and GVI‐11, respectively, which indicated that they could serve as specific delivery system for antiviral therapies. The targets of aptamers GVI‐1, GVI‐7 and GVI‐11 on the surface of GCRV‐infected cells could be membrane proteins, which were trypsin‐sensitive. Furthermore, FAM‐labelled aptamer GVI‐7 could be applied to detect GCRV infection in vivo. It is the first time to generate and characterize aptamers against GCRV‐infected cells. These aptamers have great potentials in development of rapid diagnosis technology and antiviral agents against GCRV infection in aquaculture.  相似文献   

Effects of dietary lipid level on growth performance,body composition and digestive enzymes activity of juvenile sea cucumber,Apostichopus japonicus          下载免费PDF全文

Ming‐Ling Liao  Tong‐Jun Ren  Wei Chen  Yu‐Zhe Han  Chen‐Min Liu  Zhi‐Qiang Jiang  Fu‐Qiang Wang 《Aquaculture Research》2017,48(1):92-101

A feeding trial aimed to determine the effects of dietary lipid level on growth performance, body composition and digestive enzymes activity of juvenile sea cucumber, Apostichopus japonicus. Diets with six crude lipid levels (1.9, 13.8, 29.1, 43.6, 59.6 and 71.6 g kg^?1) were fed to sea cucumbers (initial weights 0.65 ± 0.01 g) at a density of 30 juveniles, once a day. After 60 days, body weight gain (BWG), specific growth rate (SGR), feed intake (FI) and protein efficiency ratio (PER) decreased with increasing dietary lipid levels. The sea cucumbers fed 1.9 g kg^?1 crude lipid showed significantly higher (P < 0.05) BWG than those of the sea cucumbers fed 59.6 and 71.6 g kg^?1 crude lipid. Intestinal protease and lipase activities generally increased with increasing dietary lipid levels. Eicosapentaenoic acid (EPA) content of body walls generally increased with increasing dietary lipid levels. Docosahexaenoic acid (DHA) content of body walls reached the maximum value at a dietary lipid level of 13.8 g kg^?1. N‐3 highly unsaturated fatty acid content followed the same pattern of DHA. According to the growth performance and body composition of sea cucumbers, it can be indicated that the optimum dietary lipid level for juvenile sea cucumbers is between 1.9 and 13.8 g kg^?1.  相似文献   

三种免疫添加剂对草鱼非特异性免疫功能的影响   总被引：2，自引：0，他引：2  

于翔  李赫  宋文华  富丽静  姚福向  胡宗云  张涛  张健 《水产养殖》2011,32(7):38-43

在饲料中添加不同剂量的Vc、酵母多糖和壳聚糖,连续投喂42d,并在试验的第14、28、35和42天取样检测,研究其对草鱼血清溶菌酶活性、血清超氧化物歧化酶活性（SOD）活性、血清碱性磷酸酶（AKP）活性、补体C3、含量丙二醛（MDA）含量和血液白细胞吞噬活性共6种指标的影响。结果显示：实验组与对照组相比,饲料中添加Vc可以提高血清溶菌酶活性、SOD活性、AKP活性、补体C3含量和血液白细胞吞噬活性高,并降低MDA含量。添加酵母多糖可以提高血清溶菌酶活性、AKP活性、补体C3含量和血液白细胞吞噬活性,并降低MDA含量,但不能提高SOD活性。添加壳聚糖可以提高血清溶菌酶活性、SOD活性、AKP活性、补体C3含量和血液白细胞吞噬活性,但不能降低MDA含量。饲料中添加Vc、酵母多糖和壳聚糖均对草鱼的非特异性免疫功能具有一定的促进作用。  相似文献   

斑马鱼早期胚胎发育囊胚sphere时期的蛋白组学研究     

陈漪  胡瑞芹  冉皓宇  陈良标 《海洋渔业》2019,41(1):53-64

斑马鱼(Danio rerio)早期胚胎发育囊胚时期是胚胎发育过程的关键时期,利用i TRAQ蛋白质质谱分析技术,检测斑马鱼早期胚胎发育过程中囊胚sphere时期的蛋白质表达情况,并分析该时期表达的蛋白质的相应功能和参与调控的生物过程。以野生型斑马鱼发育至sphere时期即4 hpf的胚胎为样本,利用i TRAQ标记与LC-MS/MS串联质谱技术,结合数据库比对,对该时期表达的蛋白质进行定性和定量的鉴定分析。检测结果共鉴定到的总蛋白数为1 178个,利用生物信息学进行功能分析,发现这些蛋白广泛参与了细胞信号传递、细胞运动和细胞骨架构建、细胞增殖、细胞分化、物质合成与代谢等各项重要的生命活动过程。研究表明,利用i TRAQ标记的方法可以对4hpf时期的斑马鱼胚胎中的蛋白质进行有效的分离和鉴定,并初步建立了斑马鱼早期发育关键阶段sphere时期的蛋白质组表达图谱,以期为斑马鱼胚胎发育过程中蛋白质组学和调控机制的研究提供参考。  相似文献   

Replacement of fish oil with soybean oil in diets for juvenile Chinese sucker (<Emphasis Type="Italic">Myxocyprinus asiaticus</Emphasis>): effects on liver lipid peroxidation and biochemical composition     

Deng-Hang Yu  Jia-Zhi Chang  Gui-Fang Dong  Jun Liu 《Fish physiology and biochemistry》2017,43(5):1413-1420

This study was designed to evaluate the effect of the replacement of fish oil (FO) by soybean oil (SO) on growth performance, liver lipid peroxidation, and biochemical composition in juvenile Chinese sucker, Myxocyprinus asiaticus. Fish (13.7 ± 0.2 g) in triplicate were fed five experimental diets in which 0% (FO as control), 40% (SO40), 60% (SO60), 80% (SO40), and 100% (SO100) FO were replaced by SO. The body weight gain of fish fed SO40, SO60, or SO80 diet was similar to FO group, but diets that have 100% soybean oil as dietary lipid significantly reduced fish growth (P < 0.05). Although the level of SO resulted in increasing crude lipid content of the liver, the level of SO did not significantly alter the hepatosomatic index (HSI). Indicators of peroxidation, such as vitamin E (V_E) and thiobarbituric acid-reactive substance (TBARS) contents, were changed as increasing dietary SO. It was shown that the inclusion of SO in the diets increased V_E concentrations, but reduced TBARS in the liver and total cholesterol (T-CHO) in the plasma. Linoleic acid (LA) and linolenic acid (LNA) significantly increased in fish liver fed diets that contained SO, but eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and the ratio n-3/n-6 were significantly reduced by the inclusion of dietary SO (P < 0.05). Our results indicated that the inclusion of SO increased the hepatic V_E content and reduced lipid peroxidation in fish. However, diet containing 100% SO as dietary lipid could reduce growth performance. Thus, we recommended that 40–80% SO can be used as dietary lipid to replace FO for juvenile Chinese sucker.  相似文献   

南昌月亮湖夏季浮游动物昼夜垂直分布及迁移特征     

胡旭仁  聂雪  兰志春  金斌松  秦海明 《水生态学杂志》2016,37(5):23-32

2013年8月1日19∶00至2日18∶00,通过间隔1 h的定量采样,研究南昌市典型城市湖泊月亮湖中浮游动物(轮虫、枝角类和桡足类)的昼夜垂直分布规律。结果表明,月亮湖共发现浮游动物20种,其中轮虫16种,枝角类1种,桡足类3种;剪形臂尾轮虫(Brachionus forficula)、桡足类无节幼体(Copepoda Nauplii)和迈氏三肢轮虫(Filinia maior)为优势物种,分别占总捕获数量的48.27%、15.75%和11.11%。浮游动物主要类群和优势物种的Morisita指数均大于1,说明月亮湖中的浮游动物群落呈聚集分布。密度垂直分布和平均滞留深度分析结果显示,浮游动物垂直分布昼夜差异明显,夜间(19∶00-05∶00),浮游动物先聚集于月亮湖中层,01∶00后逐渐迁移到表层;白天(06∶00-18∶00),浮游动物逐渐从表层向中层和底层迁移;主要类群中轮虫、枝角类和桡足类的迁移均表现为夜间上升、白天下降、傍晚再上升,轮虫和桡足类的迁移幅度小于枝角类。优势种剪形臂尾轮虫和无节幼体的昼夜垂直迁移表现为夜升昼降,至傍晚再上升;迈氏三肢轮虫昼夜垂直分布差异显著(P0.05),但昼夜垂直迁移活动不明显。相关分析表明,水温、p H值和溶解氧对月亮湖浮游动物的垂直分布和迁移活动有显著影响。  相似文献   

Effect of Chitosan on Intestinal Bacteria of Allogynogenetic Crucian Carp,Carassius auratus gibelio,as Depicted by polymerase chain reaction‐denaturing Gradient Gel Electrophoresis     

Xuemei Li  Yuhe Yu  Shouqi Xie  Qingyun Yan  Yuhang Chen 《Journal of the World Aquaculture Society》2011,42(4):539-548

A 16S rDNA‐based polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE) method was applied to detect intestinal bacterial communities of juvenile allogynogenetic crucian carp, Carassius auratus gibelio, fed with chitosan‐containing diet. This is the first time to use the molecular method to analyze the bacterial communities in the allogynogenetic crucian carp intestine. The DGGE profile with universal bacterial primers revealed simple communities in all treatment groups. Sequencing and phylogenetic analysis of excised DGGE bands showed that the dominant bacteria belonged to class γ‐Proteobacteria and Fusobacteria. The relative abundance and diversity of detected bacteria suggested that 0.5 and 0.75% of chitosan in diet were optimum for juvenile allogynogenetic crucian carp. As in these concentrations, some detected pathogen bacteria either disappeared or decreased. However, the DGGE profile with Aeromonas‐specific primers showed a similar composition among all treatment groups, which suggested that Aeromonas was one of relative stable bacteria components in the intestine of juvenile allogynogenetic crucian carp.  相似文献   

内蒙古达里诺尔湖水质现状调查   总被引：3，自引：0，他引：3  

张玉  王洪滨  何江  刘海涛  朱存良  王志新  张建明  赵月红 《水产科学》2008,27(12)

通过应用各种物理及化学仪器分析了达里诺尔湖水温、酸碱度、溶解氧、各种离子、总含盐量等因子,根据以往水质调查结果及这次测定数据比较,达里诺尔湖的pH有不断上升的趋势,这将危及到雅罗鱼及水中生物的生存,而其他各种离子均有小幅度的上升,溶解氧基本能满足鱼类的生存需求.通过水质的调查结果分析,使瓦氏雅罗鱼及其他水生生物更好地发挥在达里诺尔湖中生存的潜力.  相似文献   

莫桑比克鳗鲡养殖生物学主要特性研究   总被引：5，自引：1，他引：5  

樊海平  林煜  钟全福  廖碧钗  卓玉琛  辛志明 《福建水产》2008,(4)

对莫桑比克鳗鲡（Anguillamor morata）玻璃鳗经形态学鉴定品种后,开展了主要养殖生物学特性研究。耐受高温和低温分别为39℃和7℃,适宜生存水温15℃～33℃;耐受盐度与水温相关,水温30℃致死盐度2．9％,适宜盐度小于1．1％,20℃致死盐度2．7％,适宜盐度小于1．8％,10℃致死盐度2．1％,适宜盐度小于2．0％;耐受pH值与水温相关,30℃致死pH值分别为3．5和11．0,适宜pH值6．0—7．0,20℃致死pH值分别为4．0和11．0,适宜pH值6．0～8．0,10℃致死pH值分别为3．5和10．5,适宜pH值5．5—8．5。常用药物S高聚碘、二氧化氯、三氯异氰尿酸、苯扎溴铵、溴氯海因、甲醛、敌百虫的安全浓度高于或接近常用浓度,硫酸铜、溴氰菊脂乳油、氯氰菊脂、甲苯咪唑的安全浓度低于常用浓度。水体中非离子态氨的安全浓度为2．51mg／L,亚硝酸氮的安全浓度为55．12mg／L。  相似文献   

凡纳滨对虾28.5D血蓝蛋白的降解新片段     

章跃陵  叶向群  陈洁辉  黄通旺  胡忠  李远友 《中国水产科学》2008,15(3):425-430

血蓝蛋白是一种具有多种非特异性免疫学活性且可以产生免疫学活性降解片段的多功能蛋白。为了探索血蓝蛋白新的降解片段,本研究以凡纳滨对虾旺itopenaeus vannamei）为研究对象,采用比较蛋白质学技术分析对虾感染病原菌后血清蛋白质组的变化。结果发现,对虾感染副溶血弧菌（Hbrio parahaemolyticus）12h后,血清中新增一种28．5kD的蛋白（命名为p28．5）。经MALDI-TOF／MS分析,其与凡纳滨对虾血蓝蛋白75kD亚基具有高度同源性。进一步研究显示,p28．5蛋白不仅可与抗血蓝蛋白抗体发生特异性结合,而且还与对虾抗感染能力呈正相关。由此推测,p28．5蛋白应该是对虾感染病原菌后产生的一种28．5kD血蓝蛋白降解新片段,其可能是血蓝蛋白发挥免疫学功能的一种新方式的产物。[中国水产科学,2008,15（3）：425-430]  相似文献