Prokaryotic Expression and Antigenicity Analysis of Porcine Japenese Encephalitis Virus Envelope Protein Domain Ⅲ

In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×10⁵ anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×10⁴ anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV.     

GM1 gangliosidosis in a Japanese domestic cat: a new variant identified in Hokkaido,Japan

A male Japanese domestic cat with retarded growth in Hokkaido, Japan, showed progressive motor dysfunction, such as ataxia starting at 3 months of age and tremors, visual disorder and seizure after 4 months of age. Finally, the cat died of neurological deterioration at 9 months of age. Approximately half of the peripheral blood lymphocytes had multiple abnormal vacuoles. Magnetic resonance imaging showed bisymmetrical hyperintensity in the white matter of the parietal and occipital lobes in the forebrain on T2-weighted and fluid-attenuated inversion recovery images, and mild encephalatrophy of the olfactory bulbs and temporal lobes. The activity of lysosomal acid β-galactosidase in leukocytes was negligible, resulting in the biochemical diagnosis of GM1 gangliosidosis. Histologically, swollen neurons characterized by accumulation of pale, slightly granular cytoplasmic materials were observed throughout the central nervous system. Dysmyelination or demyelination and gemistocytic astrocytosis were observed in the white matter. Ultrastructually, membranous cytoplasmic bodies were detected in the lysosomes of neurons. However, genetic analysis did not identify the c.1448G>C mutation, which is the single known mutation of feline GM1 gangliosidosis, suggesting that the cat was affected with a new variant of the feline disease.     

花鲈虹彩病毒交叉引物恒温扩增检测方法的建立

【目的】花鲈虹彩病毒(Lateolabrax maculatus iridovirus,LMIV)严重威胁花鲈养殖业安全,无特效防控药物,早期诊断在LMIV防控中发挥极其重要的作用。建立一种简便、快捷、准确的现场快速诊断方法,可为LMIV的基层诊断提供技术支撑。【方法】利用交叉引物恒温扩增技术(Cross priming amplification,CPA),针对LMIV ATPase基因高保守区设计1套单交叉引物。以构建的ATPase重组质粒作为阳性模板,对反应体系中的引物浓度比,Bst DNA聚合酶、Betaine、MgSO₄、dNTP浓度,以及反应温度和反应时间进行优化;结合一次性核酸试纸条,建立可视化检测LMIV-CPA的方法。【结果】最优引物浓度比组合为交叉引物CPF1.0μmol/L,引物F3和B3均为0.4μmol/L,探针引物B1(FAM)和B2(Biotin)均为0.8μmol/L;MgSO4浓度为6 mmol/L、Betaine浓度为0.4 mol/L、dNTP浓度为0.6 mmol/L、Bst DNA聚合酶浓度为0.256 U/μL;最佳反应...     

刺参与日本对虾池塘高效混养技术

刺参夏眠期间正是日本对虾生长的大好季节,将刺参和日本对虾混养,在7月份放养虾苗,严格控制养殖池内的杂鱼虾等,就可大大提高日本对虾和刺参的成活率,利用此二者在饵料和空间的互补性,可显著提高虾池的利用率和经济效益。     

Fluorescence localization of epidermal cathepsins L and B in the Japanese eel

Histochemical localization of proteolytic activities in the dorsal epidermis of Japanese eel was demonstrated by fluorescent microscopy utilizing 4-methoxy-2-naphthylamide (4M$\beta$NA) derivatives as substrates and 5-nitrosalicylaldehyde as a trapping agent. Carbobenzoxy-L-phenylalanyl-L-arginyl-4M$\beta$NA (Cbz-Phe-Arg-4M$\beta$NA) and Cbz-Arg-Arg-4M$\beta$NA were used for direct detection of cathepsins L and B activities, respectively, in fresh frozen sections and unfixed cells of the eel epidermis. The fluorescing areas, where Cbz-Phe-Arg-4M$\beta$NA was hydrolyzed by cathepsin L, were shown in mucus secretory cells and club cells and broadly around skin surface. The fluorescing areas due to Cbz-Arg-Arg-4M$\beta$NA hydrolysis by cathepsin B were localized similarly in these tissues. The fluorescing intensity for both catheptic activities in mucus secretory cells was higher than that in club cells, where small fluorescing granules were distributed. These results indicate that eel cathepsins L and B are stored in epidermal secretory cells at different levels and probably serve as defense factors before or after secretion by these cells. Abbreviations: Cbz – carbobenzoxy; 4M$\beta$NA – 4-methoxy-2-naphthylamide; NSA – 5-nitrosalicylaldehyde.     

Survival and serum biochemical responses of spotted sea bass Lateolabrax maculatus during simulated waterless live transportation

Waterless live fish transportation is an alternative and promising transport strategy, which may reduce shipping costs, increase stocking density and improve survival of some species. Spotted sea bass is one of the most economically important marine fish in China. However, waterless transportation has not yet been studied on spotted sea bass. To explore the possibility of waterless transportation of spotted sea bass and its sublethal stress responses, the survival rate and dynamics of blood biochemical parameters were evaluated in the present study by preserving it under no water, less water and water (control) conditions for 9 hr. The serum biochemical parameters including total protein, cortisol, glucose, lactate, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total carbon dioxide (TCO₂), creatinine, blood urea nitrogen (BUN), ammonia, superoxide dismutase (SOD) activity and malonaldehyde (MDA) were investigated. Results showed that the survival rates of fish in all groups gradually declined with time increasing. At the end of transportation, the survival rates in the control group, less‐water group and no‐water group decreased to 49.11%, 15.60% and 23.96% respectively. Significant changes were observed in several selected serum biochemical parameters, including glucose, lactate, ALT, TCO₂, BUN and ammonia during simulated waterless transportation, which might negatively affect the physiological homeostasis of fish, leading to the high mortality in present study. In general, waterless transportation of spotted sea bass was possible, but it might be rather suitable for short time than long time.     

日本渔业概况

日本是全球重要的渔业国家。本文介绍了日本的渔业生产、渔业资源、渔业在国民经济中的地位、渔业管理以及渔业劳动力和经营状况。日本渔业以海洋捕捞为主,由于全球渔业资源衰退,国际竞争加剧以及日本国内经济的持续低迷,日本渔业在经历了上世纪80年代的鼎盛期之后呈现大幅下降趋势,面临着产业结构调整的巨大压力。     

2种LED灯的蓝圆鲹和竹荚鱼渔获比较

为了探索节能减排的海洋捕捞作业方式,在中国沿海海域做了白光（偏绿光）发射二极管（LED）水下灯和蓝紫光LED水下灯集鱼试验。对比2种等光能LED集鱼灯周围蓝圆鲹（Decapterus maruadsi）和竹筴鱼（Trachurus japonicus）的渔获,对渔获数据进行t检验分析表明,白光LED灯对蓝圆鲹的光诱效果显著优于蓝紫光LED灯;白光LED灯对竹筴鱼的光诱效果极显著优于蓝紫光LED灯。海上试验结果证实,在实验室水槽中所得的光诱鱼试验结果对海洋捕捞实践具有很好的指导作用。     

浙北地区利用冬闲虾塘饲养日本对虾的试验

利用饲养南美白对虾的冬闲虾塘 ,采取精养模式养一茬日本对虾 ,可提高围塘养殖的综合效益。本试验的放养密度为 2× 10 4余尾 / 6 6 7m2 ,单产 38.5kg/ 6 6 7m2 ,成活率 2 3.3%。盐度控制在 17以上     

Adapting bioenergetic factorial modelling to understand the implications of heat stress on barramundi (Lates calcarifer) growth,feed utilisation and optimal protein and energy requirements – potential strategies for dealing with climate change?

The implications of temperature on bioenergetics for barramundi (Lates calcarifer) were defined in an improved factorial model that encompassed revised parameters accounting for effects over the temperature range of 16–39 °C and size range of 10–3000 g. A revised growth function describing weight gain by barramundi as a function of fish weight and temperature was derived from farm and laboratory data and included a term for a shift in optimal temperature with fish size: Gain (g fish^?1 day^?1) = (K + xT + yT ² + zT³) * (weight)^ax+b. Maintenance energy and protein demand functions were also derived on a similar form, and all three functions combined to form the basis of a factorial model for energy and protein demand. Using this model, optimal iterative feed specifications were defined for a range of fish sizes at temperatures of 25, 30 and 35 °C. A feed demand model was also developed based on the demand for digestible energy (DE) at each of these temperatures. The model shows that at high temperatures (35 °C), there is an increase in digestible protein (DP) to DE demand, and that with increasing size, there is a decrease in the DP to DE demand.