Occurrence and Effects of the Rhizocephalan Parasite,Polyascus gregarius,in the Chinese Mitten Crab,Eriocheir sinensis,Cultured in a Freshwater Pond,China

An epizootic outbreak of the parasitic barnacle, Polyascus gregarius, was reported for the first time in cultured commercial mitten crab, Eriocheir sinensis. Observations were made in a freshwater culture pond in Shenjia Dang, Zhejiang province, China, from June to October 2007. The surface morphology and appearance of the interna, externa, or scar of P. gregarius were recorded in 1464 crabs, including 871 males and 593 females. Overall prevalence of infection was 37.8 and 31.7% in male and female crabs, respectively. The monthly variation of infection was mainly dependent on the mortality of the host, with the highest prevalence (58.0% of males and 51.3% of females infected) in June, and the lowest (13.4% of males and 6.3% of females) in October. A host with multiple externae or scars was encountered frequently, with the maximum of 28 externae or 19 scars on a single crab. Being restricted in the culture pond, the hosts could not escape and return to the brackish water of the estuary; therefore, the externae were never exposed to male cyprids and remained unfertilized. As a result, all the externae degenerated and dropped off finally after 2–3 mo. This parasite caused castration, anecdysis, and increased mortality in the host, and therefore, had both direct and indirect effects on the commercial culture of E. sinensis. Infected males were castrated and displayed morphological modifications of secondary sexual characters. The size of infected crabs was significantly smaller than that of uninfected crabs because the molting was reduced by infection. After all the externae dropped off, the scarred host crabs finally died. A high mortality (possibly >50%) due to parasitism was observed in this study.     

溶珊瑚弧菌hapR基因的敲除及功能研究

溶珊瑚弧菌(Vibrio coralliilyticus)是多种珊瑚的致病菌。hapR是弧菌群体感应系统核心的泛调控基因。前期实验中发现，溶珊瑚弧菌在感染佳丽鹿角珊瑚时，致病力与其接种密度密切相关，推测溶珊瑚弧菌的群体感应系统可能是调控其毒力的关键因子。笔者利用同源重组技术成功构建了溶珊瑚弧菌hapR缺失株△hapR-Vc450和回补株ChapR-Vc450，表型测试结果表明，hapR的缺失影响溶珊瑚弧菌的菌落形态，但不影响细菌的运动性和鞭毛结构；hapR缺失后，细菌最大生长量明显减少，但生物膜的形成量显著增加；用溶珊瑚弧菌人工感染鹿角珊瑚，结果显示，hapR缺失后菌株毒力增加，表明hapR在溶珊瑚弧菌中可能具有与霍乱弧菌hapR相似的功能，可以抑制毒力基因的级联激活。     

Biphenyls from Nicotiana tabacum and their anti-tobacco mosaic virus

Five new biphenyls, tababiphenyls A–E (1–5), together with five known ones (5–10), were isolated from the leaves of Nicotiana tabacum, of which compound 1 possessed a seldom reported 6-carbons unit in biphenyls. Their structures were established on the basis of extensive spectroscopic analyses. All compounds were tested for their anti-tobacco mosaic virus (anti-TMV) activities. The results showed that compounds 3 and 5 exhibited high anti-TMV activities with inhibition rate of 48.4% and 32.1%, respectively, which were higher than that of positive control (ningnanmycin). The other compounds also showed potential anti-TMV activities with inhibition rates in the range of 18.6–28.7%, respectively.     

Biodegradation of 1-Methylindole and 3-Methylindole by Mangrove Sediment Enrichment Cultures and a Pure Culture of an Isolated Pseudomonas Aeruginosa Gs

Indolic compounds are N-heterocyclic aromatic chemicals and have been detected at contaminated sites. Biodegradation of 1-methylindole (1MI) and 3-methylindole (3MI) was investigated initially using enrichment cultures with mangrove sediment obtained from Mai Po Nature Reserve of Hong Kong and subsequently with a pure culture of Pseudomonas aeruginosa Gs confirmed with 16S rRNA gene. At 2.0 mM, 1MI and 3MI were degraded in 4 and 3 days, respectively, by the respective 1MI- and 3MI-degrading enrichment cultures. When substrate concentrations were increased to 3.0,mM and 3.5,mM, slower degradation of 1MI and 3MI was observed indicating inhibitory effects from the substrates, possibly due to toxicity. In addition, no colony of bacteria could be observed on the agar plates amended with 3.5,mM 1MI or 4.0,mM 3MI, indicating that 1MI was more toxic than 3MI. Pseudomonas aeruginosa Gs, isolated from the enrichment culture, effectively utilized both substrates as the sole source of carbon and energy. Complete degradation of 1MI and 3MI was achieved after more than 40 days and 24 days, respectively, at an initial concentration of 2,mM in the culture. Effects of initial substrate concentration, pH and salinity on degradation of 1MI and 3MI by P. aeruginosa Gs were also studied in batch culture. The optimum pH and salinity for degrading both substrates by P. aeruginosa Gs was 7.0 and 5?, respectively. Biodegradation kinetics of 1MI and 3MI by P. aeruginosa Gs could be described using a first-order kinetic model. Our results suggest that both 1MI and 3MI are biodegradable in the mangrove environment and that toxicity of 1MI could be a potential factor limiting the removal of the chemical in the environment by microorganisms.     

Effect of starvation on biochemical composition and gametogenesis in the Pacific oyster <Emphasis Type="Italic">Crassostrea gigas</Emphasis>

The effects of starvation on biochemical composition and gametogenesis were investigated in the Pacific oyster Crassostrea gigas. Histological analysis, combined with oocyte examination and measurements of protein, glycogen and lipid levels and RNA/DNA ratio from gonad, adductor muscle and mantle tissue of each sex were performed. In the starved groups, C. gigas showed gonad development, but the progress was delayed during the experiment. Glycogen was the first substrate used by C. gigas for dealing with lack of food. While glycogen was rapidly consumed, protein and lipid contents decreased gradually. A decrease in the RNA/DNA ratio in the starved groups in all the body components was found during starvation, illustrating that RNA/DNA ratio was a valid indicator of nutritional condition in C. gigas. A significant increase in water and ash contents and a corresponding decrease in condition index were observed in the starved groups, showing that the water and ash content and condition index were related to the usage of glycogen, lipid and protein reserves in body composition. During starvation, energy reserves were mobilized for survival and gonad development, but spawning was arrested. The information obtained in this study is useful for broodstock management in the Pacific oyster industry.     

5科11种鱼类ITS1特征分析及其在系统分类研究中的适用性

为了探讨ITS1作为分子标记用于鱼类系统演化的适用性,实验选取鲈形目5科11种鱼类为研究对象,包括尖吻鲈科、射水鱼科、军曹鱼科、剑鱼科和鲹科。通过克隆和测序等技术共获得了348条ITS1序列,长度范围为442~661 bp;通过对所有序列的长度、变异位点数量、GC含量、核苷酸多样性及单倍型多样性指数等遗传特征比较分析发现,11种鱼类ITS1序列无论是在种内还是在种间,长度和序列都表现出较为明显的多态性。特别是在军曹鱼中,70条克隆的长度范围为648~661 bp,但有一条序列存在55 bp缺失,结合该序列的GC含量,二级结构和最小自由能,推断该序列为假基因。以鮣为外类群,基于核糖体ITS1序列构建的邻接树显示在物种种类水平上,不同个体的克隆都按种类聚支,ITS1可以用于该类群物种的区分;在属级水平上,ITS1将11属鱼类完全区分开,能够用于属级水平的区分;在科级水平上,虽然鲹科分为2支的分子结果和形态分类存在差异,但ITS1构建的系统关系与线粒体分子标记构建的系统进化树相似。研究表明,核糖体ITS1可以作为一种有效的分子标记用于研究鱼类的系统分类研究,并且不同的分类阶元其解析能力不同,这将为鱼类核糖体的研究提供科学依据。     

11种鲈形目鱼类ITS2特征及系统应用

ITS2 (Internal transcribed spacer 2)是位于核糖体5.8S和28S基因之间的非编码序列。为了探讨该片段的多样性特征以及进化模式,本研究选取了鲈形目(Perciformes) 5科11种鱼类为研究对象,共获得了444条ITS2克隆序列,其长度范围为332～515 bp。比较种内不同序列的长度发现,金带细鲹(Selaroides leptolepis)在种内存在24 bp的差异,剑鱼(Xiphias gladius)在种内存在32 bp的差异,这2种鱼类的差异较为明显;其余9种鱼类的长度相对比较保守,长度差异小于14 bp。依据11种鱼类的保守位点数、变异位点数、简约信息位点数、单倍型数、保守位点比例、单倍型多样性指数、核苷酸多样性等特征分析发现,种内存在着不同程度差异,特别是金带细鲹的ITS2序列存在着Type A、Type B和Type C 3种类型,各类型间差异较大。根据序列的多样性特征推断,金带细鲹和剑鱼的进化方式为非协同进化;蓝圆鲹(Decapterus maruadsi)、大甲鲹(Megalaspis cordyla)、吉打副叶鲹(Alepes djedaba)和日本竹筴鱼(Trachurus japonicas)的长度和变异位点均存在着一定程度的差异,但差异并不明显,视为不严格的协同进化;泰拉鰆鲹(Scomberoides tala)、布氏鲳鲹(Trachinotus blochii)、尖吻鲈(Lates calcarifer)、射水鱼(Toxotes chatareus)和军曹鱼(Rachycentron canadum) 5种鱼类为协同进化;另外,协同和非协同进化状态与分类系统没有相关性。序列比对发现,大甲鲹种内存在着由协同进化方式演变为非严格的协同进化方式的过度序列;在金带细鲹的3个不同个体中,序列间存在着从协同进化、非严格的协同进化演变为非协同进化的3种进化方式。基于ITS2序列构建的11种鱼类的邻接系统树显示,每种鱼类的克隆都分别按种单独聚为一支,鲹科7属鱼类各属也是单独聚支,表明ITS2不仅可以用在种类的分子鉴定,同时也可以作为分子标记应用于鲹科和属级水平的系统关系研究。     

Identification and characterization of a novel lymphocystis disease virus isolate from cultured grouper in China

Grouper Epinephelus spp. is one of the most important mariculture fish species in China and South-East Asian countries. The emerging viral diseases, evoked by iridovirus which belongs to genus Megalocytivirus and Ranavirus, have been well characterized in recent years. To date, few data on lymphocystis disease in grouper which caused by lymphocystis disease virus (LCDV) were described. Here, a novel LCDV isolate was identified and characterized. Based on the sequence of LCDV major capsid protein (MCP) and DNA polymerase gene, we found that the causative agents from different species of diseased groupers were the same one and herein were uniformly defined as grouper LCDV (GLCDV). Furthermore, H&E staining revealed that the nodules on the skin were composed of giant cells that contained inclusion bodies in the cytoplasm. Numerous virus particles with >210 nm in diameter and with hexagonal profiles were observed in the cytoplasm. In addition, phylogenetic analysis based on four iridovirus core genes, MCP, DNA polymerase, myristoylated membrane protein (MMP) and ribonucleotide reductase (RNR), consistently showed that GLCDV was mostly related to LCDV-C, followed by LCDV-1. Taken together, our data firstly provided the molecular evidence that GLCDV was a novel emerging iridovirus pathogen in grouper culture.     

海水贝类养殖中的问题及对策

正近几年,海水养殖业规模迅速扩大,产量大幅度增加,养殖形势及前景良好。继藻类和虾类养殖产业发展高潮之后,贝类养殖已发展为世界最大规模的养殖产业,对沿海经济的发展发挥着重大作用。但由于养殖户发展养殖生产的积极性无度发挥,片面追求高产,重数量、轻管理,过多地消耗资