β-1,3-葡聚糖及其羧甲基衍生物对海湾扇贝免疫功能的影响

2003年3月和2004年4月，新疆某养殖场和西安灞桥区某养殖场的白斑狗鱼（Esox lucius Linnaeus）相继爆发“败血症”。从具有明显症状的濒死病鱼体表病灶和内脏组织分离得到12株优势菌株，经人工感染试验表明：其中10株分离菌为致病菌。对分离菌株进行形态、培养特征、生理生化试验鉴定，认为该病原菌为杀鲑气单胞菌史氏亚种（Aeromonas salmonida subsp．smith）。血清学试验表明各分离菌株具有相同的保护性抗原。组织病理学研究表明：患病狗鱼病灶肌纤维明显肿胀断裂，肌纤维束之间出现间隙；肝胰组织严重坏死，细胞核溶解消失，细胞浆崩解；肾小管绝大部分均已坏死，细胞破裂，胞浆呈红色团块状流失于管腔。脾脏细胞大量坏死，髓质内含铁血黄素沉积。代表菌株EL0304-1药敏试验结果为：对供试24种抗菌药物中的磺胺二甲嘧啶钠、磺胺二甲嚯唑钠等不敏感；庆大霉素、卡那霉素等中度敏感；对氧氟沙星、培氟沙星、新霉素等高度敏感。     

白鲢出血病病原菌的鉴定

从患出血病的白鲢体内分离出两株细菌(编号为水_1、水_2),分别感染健康白鲢,人工复制白鲢出血病获得成功。对这两株细菌进行形态、营养需求、生长特性、以及生化反应等观察,作者认为,水_1菌为产碱假单胞菌(Pseudomonas alcaligenes);水_2菌为嗜水气单胞菌(Aeromonas hydrophila)。前者隶属于假单胞菌科(Pseudomonaceae),后者隶属于孤菌科(Vibrionaceae)。     

中国海洋生物多样性保护研究进展与几个热点问题

根据近年最新研究成果,对中国海洋生物多样性保护和研究进展进行了综述。文章重点阐述了海洋生物遗传多样性、物种多样性和生态系统多样性状况、保护现状及其研究进展,分析了目前海洋生物多样性保护所面临的主要威胁;探讨了近期中国海洋生物多样性保护研究的几个热点问题。     

锦鲤暴发性出血症病原的研究

从患暴发性出症的锦鲤体内分离出2株细菌－052202和052203，应用细菌分离鉴定方法和法国Biomerieux FRC公司的API全自动微生物鉴定系统进行鉴定，结果显示：052202株为嗜麦芽窄食单胞菌（Stenotrophomonas maltophilia),052203株为温和气单胞（菌（Aeromonas sobria)。生物毒性试验的结果表明，温和气单胞菌可使健康锦鲤死亡，引起体表出血，肝，肾，脾，胆囊发生病变，有致病性；而嗜麦芽窄食单胞菌不引起健康锦鲤死亡，无致病性，28例药物的药敏试验结果表明，温和气单胞菌对新霉素，卡那霉素，庆大霉素，头孢噻吩，头孢曲松的头孢他啶的敏感性强或较强。     

Aeromonas sobria, a causative agent of disease in farmed perch, Perca fluviatilis L

Significant numbers of perch, Perca fluviatilis, raised on a pilot fish farm in Switzerland presented focal skin lesions on the lateral sides and fin rot. Mortality rates reached levels of up to 1% of the total fish on the farm per day. Virtually pure cultures of Aeromonas sobria were isolated from the liver, kidney, spleen and skin lesions of affected fish. Aeromonas sobria isolated from the farmed perch had a haemolytic effect on sheep and trout erythrocytes, autoaggregated, was cytotoxic for cultured fish cells and possessed genes involved in type III protein secretion. Experimental infection of naive perch with a single colony isolate of A. sobria from an affected farm fish resulted in the development of clinical signs identical to those seen on the farm. The results indicate that A. sobria can act as a primary pathogen of perch.     

Analysis of exotoxins produced by atypical isolates of Aeromonas salmonicida,by enzymatic and serological methods

In this study, exotoxins produced by 62 Aeromonas salmonicida strains and the bacterium Haemophilus piscium were analysed. Enzymatic assays, zymograms and serological detection were used to monitor secretion by bacterial strains of the previously described exotoxins P1, GCAT and AsaP1 and also the extracellular P2 metallo-gelatinase and a serine caseinase, which is different from the P1 protease and has not yet been characterized. Based on the results, the strains were divided into five groups. One comprised the type strains for A. salmonicida ssp. masoucida, H. piscium and 36% of the atypical isolates, and another, a type strain for A. salmonicida ssp. smithia together with 14% of the atypical isolates. A second type strain of A. salmonicida ssp. smithia was grouped with 8% of the atypical isolates. The largest group contained the type strains for A. salmonicida ssp. achromogenes and 38% of the atypical isolates. The type strains for A. salmonicida ssp. salmonicida were in the last group with all the four typical strains and 4% of the atypical isolates. The combination of zymogram and serological detection used is recommended as the most reliable method for characterizing A. salmonicida strains according to their exotoxin secretion.     

海水网箱养殖对近岸环境影响的研究进展

近年来,我国的海水网箱养殖业发展迅猛,已成为我国渔业新兴产业的生力军。海水网箱养殖系统作为一种高密度、高投饵的人工养殖生态系统,其输出的废物,残饵、代谢及排泄废物等是引发环境问题的主要污染源,再加上网箱养殖区布局不合理,通常设在水交换率较低的内湾,当养殖容量超出了海域的环境容量,就会引发一系列生态环境问题[1]。本文就海水网箱养殖对环境污染及防治的研究进展进行综述,并提出今后的重点研究方向。1养殖区水质污染网箱养殖对浅海生态系统的影响主要表现为养殖水域营养负荷增加[2]。一般来说,鲑鳟鱼类的网箱养殖,饲料中总氮的…     

维氏气单胞菌菌影疫苗的构建及其溶菌动力学研究

细菌菌影(Bacterial ghosts)是通过诱导PhiX 174噬菌体中裂解基因E在革兰氏阴性菌中的表达所获得的无细胞内容物的空细菌体.它保持了与活菌相同的膜蛋白成分、天然结构、细胞表面抗原等特性,是一种新型的死菌疫苗.为优化维氏气单胞菌(A.veronii)菌影制备条件,本实验将带有庆大抗性的裂解基因E转入A.veroniiATCC35624株中,通过温度变化对其进行溶菌动力学试验,每隔30 min对菌液OD600nm及其活菌数进行测定,当OD600nm稳定不再下降时,通过扫描电镜观察裂解后菌体的形态变化.结果显示,在诱导30 min后OD600nm值开始持续下降,当其下降到最低值时,活菌检测结果表明几乎无活菌存在.本研究利用菌影形成机制构建A.veronii菌影,为新型疫苗的制备提供依据.     

鲈细菌性类结节病的病原及血液病理研究

2002年5月从患细菌性类结节病的濒死鲈体内分离到一株细菌02517-1，经人工感染试验及API 20E系统鉴定，认为该病原菌为嗜水气单胞菌(Aeromonas hydrophila)。血液生理生化指标检测显示，与健康鱼相比，患病鱼的红细胞、白细胞、血糖、蛋白质、尿素、胆固醇等显著下降，而谷丙转氨酶ALT、谷草转氨酶AST、乳酸脱氢酶LDH等血清酶类活性增强，说明经嗜水气单胞菌感染后，鲈在营养物质代谢、抗病力、消化、呼吸、排泄等方面的生理功能受到严重损害。药敏实验结果表明：复方新诺明、卡那霉素、庆大霉素、氟哌酸等4种化学疗剂及地榆、乌梅、木瓜、五味子等4种中草药对该种病原菌有较强的抑菌作用。     

鳜嗜水气单胞菌GYK1株胞外产物提取、纯化及生物学活性分析

以鳜分离的嗜水气单胞菌GYK1株为代表,采用玻璃纸覆盖技术提取了8株嗜水气单胞菌和1株温和气单胞菌的胞外产物,SDS-PAGE电泳分析显示,8株嗜水气单胞菌的胞外产物均有35kDa的共同蛋白带。酶活性和溶血性分析表明,GYK1株胞外产物具有酪蛋白酶、淀粉酶、脂肪酶、明胶酶活性,但不具备脲酶活性;溶血性较强,蛋白浓度为365μg·mL-1的胞外产物对鳜血细胞的溶血价为213,对加州鲈、银鲫、小白鼠、兔、绵羊、人O型血的红细胞的溶血价在211~214之间。应用聚丙烯酰胺葡聚糖凝胶柱层析,对GYK1株的胞外产物进行了初步纯化,得到分子量为35kDa的蛋白带,该纯化产物具溶血性,但溶血性比粗胞外产物降低,对鳜和小鼠血细胞的溶血价均为25。