网箱养殖罗非鱼综合症的病理组织学研究

于1989至1992年对水库网箱养殖罗非鱼综合症各发病期、各症状型的病鱼进行了病理组织学研究。研究结果表明:网箱养殖罗非鱼综合症的病理基础是肝肾功能障碍,病鱼的肝、肾、脾、肠、鳃、性腺、脑均有不同程度的坏死或变性。本文还对细菌继发性感染,各症状型的病理学变化及防治对策作了讨论。     

Cross-shelf distribution of copepods and the role of event-scale winds in a northern California upwelling zone

Cross-shelf distribution and abundance of copepod nauplii and copepodids were measured during three summer upwelling seasons (2000–2002) in a coastal upwelling zone off northern California. These 3 years varied considerably in the intensity of winds, abundance of chlorophyll, and water temperature. The cruises in 2000 were characterized by relaxation conditions, with generally high levels of chlorophyll and high water temperature. The cruises in 2001 and 2002 were dominated by strong and persistent upwelling events, leading to lower chlorophyll and water temperatures. The copepod assemblage was dominated by Oithona spp., Acartia spp. and Pseudocalanus spp., with Metridia pacifica (lucens), Microsetella rosea, Oncaea spp. and Tortanus discaudatus also common during all 3 years. The cross-shelf distribution of copepods was generally shifted offshore during upwelling and onshore during relaxation events, although some variability between species occurred. Abundance of all life stages generally exhibited a negative correlation with cross-shelf transport averaged over at least 1–4 days and lagged by 0–3 days, indicating lower abundances during and immediately after active upwelling. However, copepod nauplii seemed to respond positively to wind events lasting 1–5 days followed by a period of relaxation lasting 6 or 7 days. These rapid rates of change in abundance are probably too great to be due to in situ growth and reproduction alone; physical processes must also play a role. These results suggest a highly dynamic relationship between copepods and upwelling events off northern California, with species-specific responses to upwelling to be expected.     

溶藻弧菌asp基因在大肠杆菌中表达活性研究及条件优化

为进一步研究溶藻弧菌碱性丝氨酸蛋白酶(ASP)蛋白生物学活性和功能,将构建的pET23d-asp在大肠杆菌BL21(DE3)中表达,对表达产物进行纯化分析,并优化表达条件。结果表明:在咪唑洗脱缓冲液浓度为100 mmol/L时,表达的可溶性ASP被大量洗脱,纯化的ASP相对分子质量约为42 000,具有蛋白活性;在诱导温度28℃、异丙基-β-D硫代半乳糖苷(Isopropyl-β-D-thiogalactopyranoside,IPTG)浓度1 mmol/L及诱导时间16 h时,表达的活性ASP蛋白最多。     

青蛤(Cyclina sinensis)IKK基因的克隆及其在鳗弧菌(Vibrio anguillarum)刺激下的表达分析

利用构建的青蛤(Cyclina sinensis)转录组文库,筛选到青蛤IKK基因的类似序列。经设计引物克隆比对后确认为CsIKK基因。利用生物信息学软件在线对该基因进行结构分析。采用PCR技术克隆基因,并使用实时荧光定量PCR技术克隆得到CsIKK基因在青蛤五个不同组织中的表达情况及在鳗弧菌(Vibrioanguillarum)的刺激下IKK基因在青蛤血淋巴中的时序性表达情况。综合结果得到, CsIKK基因序列开放阅读框长2298bp,编码765个氨基酸。IKK基因在青蛤的血淋巴、外套膜、闭壳肌、肝脏、性腺和鳃六个组织中均表达,在血淋巴中表达量最高。青蛤IKK基因在鳗弧菌胁迫下表达量在6h时达到最大值,与对照组相比差异极显著(P0.01),表明该基因所指导的蛋白是青蛤重要的免疫信号通路蛋白。     

凡纳滨对虾(Litopenaeus vannamei)病原副溶血弧菌(Vibrio parahaemolyticus)的表型及分子特征

采用常规的形态及生理生化特性检验及分子生物学等方法,对引起凡纳滨对虾幼虾发生毁灭性死亡的病原菌进行了致病性、表型生物学及分子生物学的系统研究。结果表明,分离菌为弧菌属(Vibrio Pacini 1954)的副溶血弧菌(Vibrio parahaemolyticus),对凡纳滨对虾、中国对虾及日本对虾仔虾均有强致病性。分离鉴定的4株副溶血弧菌具有淀粉酶、明胶酶、卵磷脂酶活性,但不具有蛋白酶、脂肪酶活性,KP溶血均呈阴性,且均具有K抗原;代表菌株(JGB080708-1株)的16S rRNA基因序列(长度为1456bp,GenBank登录号为GQ205448)和gyrB基因序列(长度为1195bp,GenBank登录号为GQ205453)与副溶血弧菌的两种基因序列相似性均可达98%以上。病原菌的耐药性检测结果显示,对供试49种抗菌药物中的林可霉素等6种药物耐药。     

6种海洋致病性弧菌36kDa外膜蛋白特性分析

研究本着筛选和制备弧菌共同外膜蛋白亚单位疫苗,用十二烷基肌氨酸钠抽提并结合超速离心的方法提取了鳗弧菌(Vibrio anguillarum)、鱼肠道弧菌(V.ichthyoenteri)、溶藻弧菌(V.alginolyticus)、副溶血弧菌(V.parahaemolyticus)、哈维氏弧菌(V.harveyi)和创伤弧菌(V.vulnificus)6种海洋致病性弧菌的主要外膜蛋白, SDS-PAGE图谱比较发现这6种弧菌都存在36 kDa的外膜蛋白条带.通过电泳洗脱分离纯化这6种弧菌的36 kDa外膜蛋白,并制备了鳗弧菌菌株SJ060621的36 kDa外膜蛋白多抗.ELISA和Western-blot印迹结果显示,仅鳗弧菌SJ060621、S010610-1和哈维氏弧菌的36 kDa外膜蛋白存在免疫交叉反应;双向电泳分析了鳗弧菌SJ060621、S010610-1、哈维氏弧菌和溶藻弧菌的36 kDa外膜蛋白,2-DE图谱显示出鳗弧菌SJ060621和S010610-1均由3种等电点相近的蛋白组成,存在很高的同源性,2株鳗弧菌与溶藻弧菌和哈维氏弧菌在蛋白质组成数量和等电点上都存在较大的差异,结果说明通过主要外膜蛋白的分子量和免疫特性研究,可为筛选不同弧菌共同外膜蛋白的亚单位疫苗提供有利参考.     

Ex situ cultivation of Aplysinaaerophoba close to in situ conditions: ecological, biochemical and histological aspects

Sponges provide the largest number of biologically active natural products known from the marine environment and continue to be a very well studied phylum of marine fauna. The Mediterranean sponge Aplysina aerophoba accumulates brominated isoxazoline alkaloids such as Aplysinamisin-1, Aerophobin-2, Isofistularin-3 and the biotransformation product Aeroplysinin-1, which possesses, for example, antibiotic and cytotoxic properties. Until now, it is still being discussed which organisms – the sponge itself or associated microorganisms – are responsible for metabolite production. For cultivating Aplysina individuals under ex situ conditions, we surveyed relevant ecological factors in situ and controlled them in our aquarium system. We maintained A. aerophoba for more than 9 months and analysed changes of metabolite content and composition, microbial association as well as morphology in situ and ex situ under different light exposure. Although sponges showed slight reduction during maintenance, ex situ cultivation similar to in situ conditions provides a promising method to keep sponges and obtain their bioactive metabolites.     

三氯生对小球藻的生长效应研究

研究三氯生(Triclosan) 对小球藻(Chlorella spp) 生长状况、藻细胞光合色素及碳水化合物含量的影响,检测三氯生对小球藻的毒性效应.采用概率单位-浓度对数法进行数据处理.结果表明:藻细胞密度随药剂浓度增加而明显下降,且呈现较好的剂量-效应关系;96 h 半抑制效应浓度 ( EC50 ) 为 0.065 0 mg·L-1,属于高毒;三氯生明显抑制小球藻光合色素含量的增加,并对碳水化合物含量变化有持续的影响.     

末次冰盛期与全新世青藏高原松属植被分布及其演变

史前人类与其生存环境,特别是与植被之间的相互作用一直是人们关注的焦点。气候变化与人类活动对植被覆盖格局的塑造均有重要影响,厘清植被分布及其植被变化背后的气候或人为因素,有助于更好地认识人类活动对植被环境的影响程度与范围。本研究对青藏高原松属（Pinus spp.）植被适生区分布动态历史进行了物种分布模型模拟,并结合环境考古学、古生态学等多学科方法,探究青藏高原松属植被分布变化历史及其驱动因素。结果表明,基于最大熵（MaxEnt）算法并结合R语言包ENMeval来优化参数设置的模型性能表现良好且稳健,较为准确模拟松属在青藏高原地区不同时期的适生区分布动态。高原上松属适生区在末次冰盛期（LGM）分布最小,仅在东缘的河谷局地;在全新世早、中期分布达到最大值,到全新世晚期部分区域分布缩小,高原东北部高度适生区向2500 m a. s. l.上下的低海拔谷地收缩,整体上与高原气候变化趋势大体一致。结合化石花粉、木炭等证据表明,全新世中晚期以前,人类对高原松属植被影响总体尚不显著;进入全新世晚期,农业技术的进步促使高原东北部人口快速增加,高原上现已发现的古遗址也高度集中在此。结合多项考古证据表明,人类对松属为代表的林木资源规模化开发行为开始显著干扰了当地植被,人类对松属等针叶林木材资源的利用可能是该地区树种向阔叶林树种演替的重要驱动因素之一。