草鱼鳃组织蛋白双向电泳技术的建立及优化     

徐湛宁  李福贵  陈杰  蒋霞云  邹曙明 《淡水渔业》2017,47(5)

为建立并优化草鱼鳃蛋白质组双向电泳的技术体系,实验将草鱼鳃经裂解处理后,通过固相p H梯度胶条进行一向等电聚焦和SDS聚丙烯酰胺凝胶垂直电泳进行蛋白分离,对不同的裂解液配方、蛋白纯化方法、IPG胶条的分离范围、上样量和染色方法等进行了探索和优化,并应用lmage Master 2D Platinum 7.0软件对电泳图谱进行了分析。结果表明:采用裂解液中添加DTT来取代Tris和TBP、选择分离范围为pH4-7的IPG胶条和150μg的主动水化上样,电泳结束后对凝胶进行硝酸银染色,获得了覆盖范围广、低背景、高分辨率、适合差异蛋白分析的双向电泳参考图谱。  相似文献   

鳙鳃凝集素的分离纯化及理化性质的研究     

盘赛昆  汤坚  顾小红  杨超 《水产科学》2009,28(12)

利用DEAE-Sepharose FF离子交换和Sephacryl S-200 HR及Superdex 200 10/300 GL分子筛层析技术,从鳙鳃组织中分离纯化到一种岩藻糖专一的凝集素,命名为GANL.在还原SDS-PAGE电泳上显示单一蛋白染色带,其亚基相对分子质量为37 kD.经Superdex 200凝胶过滤层析测得其天然相对分子质量为220 kD.GANL的中性糖含量为13.4%.因此,GANL作为一种由相同亚基组成多亚基糖蛋白.GANL对兔红细胞有专一的凝集活性,其凝集活性不依赖Ca~(2+).在被测的单糖、双糖及糖蛋白中,仅岩藻糖能抑制其凝集活性.氨基酸分析表明GANL的Asp、Glu、Leu、Val、Lys的含量较高,Cys-S的含量为0.81%.GANL的最适pH为8～9;具有很高的热稳定性,最适温度为50 ℃.  相似文献   

Amoebic gill disease in hatchery‐reared ayu,Plecoglossus altivelis (Temminck & Schlegel), in Japan is caused by Neoparamoeba perurans     

P B B Crosbie  K Ogawa  D Nakano  B F Nowak 《Journal of fish diseases》2010,33(5):455-458

  相似文献   

盐度对俄罗斯鲟幼鱼血清渗透压、离子含量及鳃丝 Na⁺/K⁺-ATP 酶活力的影响   总被引：3，自引：1，他引：2  

屈亮  庄平  章龙珍  赵峰  刘鉴毅  王妤 《中国水产科学》2010,17(2):243-251

研究了急性盐度胁迫对7月龄俄罗斯鲟(Acipenser gueldenstaedtii)幼鱼[体质量(86.1±18)g]鳃丝Na+/K+-ATP酶活力、血清渗透压和血清离子(Na+、K+、Cl-)的影响。结果表明:幼鱼从淡水直接转入盐度15、20、25水中,96h死亡率分别为72.22%、100%、100%,其他组(盐度0、5、10)无死亡。各盐度组96h血清渗透压、Na+、Cl-浓度随盐度升高而增加,且各盐度组显著高于对照组(P0.05),盐度15组最高,盐度5、10组间各指标不存在显著差异(P0.05)。盐度组血清K+离子在48h中较对照组均显著降低(P0.05),96h时有所回升但仍低于对照组,盐度15与盐度5、10相比血清K+离子有显著性差异(P0.05),盐度5、10之间无显著性差异(P0.05)。盐度组鳃丝Na+/K+-ATP酶活力呈先下降后上升变化,48h为最低,与对照组有显著性差异(P0.05),96h时盐度5、10组鳃丝Na+/K+-ATP酶活力与对照组及组间无显著差异(P0.05),盐度15与对照组及盐度5、10均有显著差异(P0.05)。96h幼鱼的等渗点为303.2mOsm·kg-1,相当于盐度10.06,而Na+及Cl-等离子点分别为146.1mmol·L-1和136.8mmol·L-1,分别相当于盐度9.02和8.95。与盐度15组相比,盐度10及其以下处理组各项检测指标变化幅度相对较小,在幼鱼渗透调节范围之内,盐度10中养殖15d后各项指标与淡水组差异较小,因此7月龄俄罗斯鲟幼鱼已具备在盐度10及以下的咸水中生活的能力,但不能耐受高于10的盐度。  相似文献   

Early stages of Margaritifera margaritifera glochidiosis in Atlantic salmon: Morphopathological characterization     

Pedro A. Castrillo  Catuxa Varela-Dopico  Paz Ondina  María Isabel Quiroga  Roberto Bermúdez 《Journal of fish diseases》2020,43(1):69-80

Freshwater mussels of the order Unionida encyst into the fish mucosa to metamorphose and complete their life cycle, causing a parasitic disease known as glochidiosis. This parasitic stage represents a bottleneck for the survival of naiads, particularly for critically endangered species as Margaritifera margaritifera; however, little is known about the events occurring during this critical stage. Therefore, this study aimed to histologically characterize the development of M. margaritifera glochidiosis in Atlantic salmon to get insight into the pathogenesis of this interaction. Fish exposed to glochidia were sampled during the first 44 days post-exposure, and organs were observed by stereomicroscopy and light microscopy. Glochidia attached to the gills by pinching the lamellar epithelium, whereupon an acute proliferative branchitis engulfed most of the larvae. However, during the first 14 days, a severe detachment of unviable glochidia occurred, associated with the presence of pleomorphic inflammatory infiltrate and epithelial degeneration. In the cases where larvae remained attached, a chronification of the lesions with none to scarce inflammation was observed. These results provide key information to better understand the complex host–parasite interaction during the early stages of glochidiosis and provide valuable information to optimize artificial rearing of naiads in conservation of threatened freshwater mussel populations.  相似文献   

Methacarn preserves mucus integrity and improves visualization of amoebae in gills of Atlantic salmon (Salmo salar L.)     

Carolina Fernandez  Dario Mascolo  Sean J. Monaghan  Johanna L. Baily  Lynn Chalmers  Giuseppe Paladini  Alexandra Adams  James E. Bron  Sophie Fridman 《Journal of fish diseases》2019,42(6):883-894

Two aqueous fixation methods (modified Davidson's solution and modified Davidson's solution with 2% (w/v) Alcian blue) were compared against two non‐aqueous fixation methods (methacarn solution and methacarn solution with 2% (w/v) Alcian blue) along with the standard buffered formalin fixation method to (a) improve preservation of the mucous coat on Atlantic salmon, Salmo salar L., gills and (b) to examine the interaction between the amoebae and mucus on the gill during an infection with amoebic gill disease. Aqueous fixatives demonstrated excellent cytological preservation but failed to deliver the preservation of the mucus when compared to the non‐aqueous‐based fixatives; qualitative and semi‐quantitative analysis revealed a greater preservation of the gill mucus using the non‐aqueous methacarn solution. A combination of this fixation method and an Alcian blue/Periodic acid–Schiff staining was tested in gills of Atlantic salmon infected with amoebic gill disease; lectin labelling was also used to confirm the mucus preservation in the methacarn‐fixed tissue. Amoebae were observed closely associated with the mucus demonstrating that the techniques employed for preservation of the mucous coat can indeed avoid the loss of potential mucus‐embedded parasites, thus providing a better understanding of the relationship between the mucus and parasite.  相似文献   

基于北斗船位数据的流刺网网次和方向提取方法研究   总被引：1，自引：0，他引：1  

张荣瀚  张胜茂  陈雪忠  樊伟  朱文斌 《海洋渔业》2019,41(2):169-178

渔船监测系统(vessel monitoring system,VMS)在渔业管理中发挥着重要作用,一直以来都受到各国及渔业组织的重视,我国近海渔船主要采用基于北斗的VMS系统。刺网是我国主要的捕捞作业方式之一,提出一种使用北斗数据提取流刺网作业网次以及网长和方向的方法。该方法使用阈值综合判别的方法判断捕捞作业的状态,通过航速、空间距离、时间间隔和航向变化的阈值判别作业时收网状态的船位点,然后根据收网状态的起始点判定放网状态的起始点。根据范例中提取的516组网次,使用程序判别的船位点和人工判断的船位点有74%相同,表明有较好的一致性,具有精度高、处理快速、实时程度高的特点。该方法可为我国渔业管理和相关研究提供新的手段,并可应用于捕捞控制、海上执法等多个领域。  相似文献   

Effect of hydrogen peroxide as treatment for amoebic gill disease in Atlantic salmon (Salmo salar L.) in different temperatures          下载免费PDF全文

Kristine Hov Martinsen  Audur Thorisdottir  Marie Lillehammer 《Aquaculture Research》2018,49(5):1733-1739

Amoebic gill disease (AGD) is a pathogenic disease in salmonids caused by Neoparamoeba perurans. Treatment of AGD infection has been through freshwater bathing of the fish. However, as the availability of fresh water is often limited, hydrogen peroxide has been introduced as an alternative treatment. This study investigated the effect of hydrogen peroxide as treatment for AGD‐infected salmon (Salmo salar L.,) at different seawater temperatures and hydrogen peroxide dosages. In total, 600 fish were challenged with N. perurans and the severity of the AGD infection was measured using a gill score scale. After challenge and disease development, the fish were distributed into 12 tanks. The treatment was performed at different seawater temperatures (8°C, 12°C, 17°C) using different hydrogen peroxide doses. Each temperature included an untreated control group. Linear models were used to analyse gill score. A significant effect of treatment was found (?0.68 ± 0.05) regardless of dose and temperature, suggesting that hydrogen peroxide was effective in treating AGD. When the model included dose, a negative linear relationship between dose and gill score was found. The study proved that treatment of AGD with hydrogen peroxide was successful, as gills partially recovered following treatment and further disease development was delayed.  相似文献   

Effect of repeated exposure to AQUI‐S® on the viability and growth of Neoparamoeba perurans          下载免费PDF全文

R J Chance  Z Allcock  C J Secombes  B Collet  C Collins 《Journal of fish diseases》2018,41(2):291-298

There have been recent efforts amongst immunologists to develop approaches for following individual fish during challenges with viral and bacterial pathogens. This study contributes to assessing the feasibility of using such approaches to study amoebic gill disease (AGD). Neoparamoeba perurans, agent of AGD, has been responsible for widespread economic and fish loss in salmonid aquaculture. With the emergence of AGD in Europe, research into infection dynamics and host response has increased. This study investigated the effect of repeat exposure to anaesthesia, a necessary requirement when following disease progression in individual fish, on N. perurans. In vitro cultures of N. perurans were exposed every 4 days over a 28‐day period to AQUI‐S^® (isoeugenol), a popular anaesthetic choice for AGD challenges, at a concentration and duration required to sedate post‐smolt salmonids. Population growth was measured by sequential counts of amoeba over the period, while viability of non‐attached amoeba in the culture was assessed with a vital stain. AQUI‐S^® was found to be a suitable choice for in vivo ectoparasitic challenges with N. perurans during which repetitive anaesthesia is required for analysis of disease progression.  相似文献   

Morphological diversity of Paramoeba perurans trophozoites and their interaction with Atlantic salmon,Salmo salar L., gills          下载免费PDF全文

J Wiik‐Nielsen  T A Mo  H Kolstad  S N Mohammad  S Hytterd  M D Powell 《Journal of fish diseases》2016,39(9):1113-1123

Amoebic gill disease (AGD) caused by the ectoparasite Paramoeba perurans affects several cultured marine fish species worldwide. In this study, the morphology and ultrastructure of P. perurans in vitro and in vivo was investigated using scanning and transmission electron microscopy (SEM and TEM, respectively). Amoebae cultures contained several different morphologies ranging from a distinct rounded cell structure and polymorphic cells with pseudopodia of different lengths and shapes. SEM studies of the gills of AGD‐affected Atlantic salmon, Salmo salar L., revealed the presence of enlarged swellings in affected gill filaments and fusion of adjacent lamellae. Spherical amoebae appeared to embed within the epithelium, and subsequently leave hemispherical indentations with visible fenestrations in the basolateral surface following their departure. These fenestrated structures corresponded to the presence of pseudopodia which could be seen by TEM to penetrate into the epithelium. The membrane–membrane interface contained an amorphous and slightly fibrous matrix. This suggests the existence of cellular glycocalyces and a role for extracellular products in mediating pathological changes in amoebic gill disease.  相似文献