双抗体夹心ELISA法检测养殖对虾病毒的研究

用纯化的长毛对虾球状病毒（PPSV）和日本对虾中肠腺坏死杆状病毒（BMNV）制备新西兰兔抗BMNV和抗PPSV抗血清及Balb／c小鼠抗BMNV和抗PPSV抗血清,建立检测PSV和BMNV的双抗体夹心ELISA检测法,结果表明,双抗体夹心ELISA法具有较高的灵敏度,可以从100μl待测组织匀浆液中检测到50ng的PPSV蛋白,以及100ng的BMNV蛋白。不同病毒抗血清无交叉反应性,用该ELISA技术检测养殖对虾和多种采自养殖虾池及其附近的近海岸生物,发现相当比例的外观正常的对虾和近海岸生物已呈阳性反应     

中国对虾球状病毒垂直传播的初步研究

应用透射电镜技术在越冬中国对虾雌性亲虾体内检测出一种球状病毒,直径为80nm左右。人工感染实验证实该病毒颗粒具有感染性。对带毒亲虾在室内隔离条件下进行了暂养催产,并对其所产的卵子、幼虫和的虾进行了隔离培育。带毒亲虾的卵子孵化率、幼虫成活率和幼虾生长速度均明显低于不带毒亲虾的子代。电镜观察发现．在带毒亲虾卵巢、各期幼虫和幼虾的细胞内存在球状病毒颗粒,在刚产的卵子的卵黄颗粒中可看到一种似球状病毒颗粒。研究表明：带球状病毒的对虾呈隐性感染,在胁迫条件下带毒幼虾可暴发病害,带毒亲虾可正常发育产卵,病毒可能通过卵巢、卵子进行垂直传播。     

养殖对虾的纤毛虫病害

纤毛虫原是养殖水体正常的生态种群,关于纤毛虫与对虾(Penaeus)的生态关系,早有过报道。七十年代初,随着对虾养殖业在世界范围内的蓬勃兴起,纤毛虫对养殖对虾的危害才逐渐地引起人们的注意。1973年,美国学者Johnson和Overstreet分别报道在池养的白对虾(Penaeus setiferus)和褐对虾(P. aztecus)体表发现大量聚缩虫(Zoothamnium sp.)附着,指出当溶解氧水平降到3—2.6ppm时,对虾死亡率很高。在这以后,美国著名无脊椎动物病理学家Lightner(1975a,1975b)继续报道他在其他对虾,例如桃红对虾(P. duorarum),蓝对虾(P.stylirostris)等发现的纤毛虫病,并对病理作了初步的分析。     

对虾病毒病害的研究进展

随着对虾养殖业的发展,其病害亦日趋突出。对虾病原的研究,特别是病毒性病原的研究,是当前我国发展对虾养殖业的重要课题,它对虾病的防治与检测提供重要的科学依据。为此,我们对近十多年来已发表的有关对虾病毒方面研究的论文和资料作一概述,希望能对同行的研究工作有所稗益。l对虾病原的研究l．且国外已发现的对虾病毒种类自1974年Cou。h【‘’报道了第一种对虾病毒以来,共发现了15种病毒或似病毒颗粒：对虾杆状病毒（BP）、对虾中肠腺坏死病毒（BMNV）、斑节对虾杆状病毒（MBV）、对虾传染性皮下与造血器官坏死病毒（IHHNV）…     

养殖对虾一种新杆状病毒的研究

应用超薄切片、负染色和蔗糖梯度超离心技术,在自然发病和人工感染的对虾肝、肠、淋巴样器官、鳃和肌肉组织中,发现并纯化了一种新杆状病毒.该病毒直径96～112nm,是已知对虾病毒中最粗的一种.中央是高电子密度的核心,外裹衣壳和囊膜,两膜之间有宽阔的间隙,这是已报道的任何一种对虾杆状病毒所没有的.纯化的病毒核衣壳表面有螺旋排列的亚单位,这也是本病毒特有的.主要靶细胞是淋巴样细胞,未在肝、肠和鳃上皮细胞中观察到,这与对虾杆状病毒无一例外地侵染某一种上皮细胞迥然不同.病毒仅在靶细胞核内增殖,不形成封入体或包涵体,它是一种迄今未曾报道的新病毒,称之为“淋巴样细胞核型杆状病毒”.     

对虾病毒病病毒和细菌合并感染的病理特点和诊断价值

应用电镜超薄切片及光学石蜡切片和环氧树脂薄片技术,观察健康对照组、疾病始发组和濒临死亡组的草虾、中国对虾、日本对虾和长毛对虾的肝胰腺和中肠,结果在３组４种对虾中均检测到病毒,只在濒临死亡组的４种对虾中发现病毒与细菌的合并感染。合并感染的病毒有ＭＢＶ、球形病毒和淋巴样细胞核型杆状病毒。细菌以弧菌为主,其病理变化表现为器官和组织的保护层受损、溶解样坏死和凝固样坏死。仅以病毒和细菌合并感染作为养殖对虾病毒病早期诊断的指标是不够的,仅可作为辅助性诊断指标。但合并感染对评估对虾病毒病的预后以指导捕捞,却有一定意义。     

对虾杆状病毒病暴发式大流行的生态机理初步研究

初步探讨中国和沿海和对虾杆状病毒病暴发式大流行的生理机理。分析了病毒病特性与对虾免疫功能的关系;病毒在虾池间的传染机制。着重说明了虾病的群体感染过程,给出模式化的群体传染模型,简要阐述了与虾病相关的社会生态经济问题。     

对虾综合养殖生态系底泥细菌的数量动态

1997年5～9月于山东海阳市黄海集团公司养虾场,用5个实验围隔研究了对虾综合养殖生态系底泥细菌的数量动态.结果表明,底泥细菌数量波动在4.10×108～12.36×108cells·g-1之间,平均为8.09×108±2.17×108cells·g-1.随着养殖时间的推移,底泥细菌数量逐渐增加.底泥细菌数量最大值出现在表层1cm之内,2～3cm以下泥层,细菌数量明显减小     

对虾白斑综合征病毒新株型WSSV-CN-Pc的毒力研究

对虾白斑综合征病毒(white spot syndrome virus,WSSV)是一种能够感染虾类并且造成其大面积死亡的环状双链DNA病毒。WSSV有多种分离株,其毒力有所差异。从克氏原螯虾(Procambarus clarkii)中分离得到1株WSSV新分离株WSSV-CN-Pc,其毒力尚不清楚。本研究采用肌肉注射和经口注射的方法,以WSSVTW型作为阳性对照,分别对克氏原螯虾(P.clarkii)和罗氏沼虾(Macrobrachium rosenbergii)进行活体实验。实验结果显示:肌肉注射WSSV-CN-Pc和WSSV-TW的克氏原螯虾均在第6天出现100%的死亡;罗氏沼虾在肌肉注射WSSV-TW后未出现死亡,但在注射WSSV-CN-Pc后的第9天死亡率达100%。经口注射WSSV-CN-Pc和WSSV-TW的克氏原螯虾均在第16天出现100%的死亡;罗氏沼虾经口注射WSSV-CN-Pc后的第19天死亡率为100%,但注射WSSV-TW的实验组并未出现死亡。结果表明,对于克氏原螯虾,WSSV-CN-Pc具有和WSSV-TW相似的毒力,而对罗氏沼虾存在明显的毒力差异。提示克氏原螯虾是WSSV传播途径中的重要因素。     

对虾养殖围隔生态系颗粒悬浮物的研究

对５个对虾养殖围隔生态系颗粒悬浮物进行了研究．结果表明,颗粒悬浮物总量波动在２０．８３～１７２．５０ｍｇ·Ｌ－１之间,平均７２．２６±３９．８７ｍｇ·Ｌ－１,其中颗粒有机物占６２％,颗粒无机物占３８％．颗粒有机物中,颗粒腐质与细菌占９８％,浮游生物占２％（其中浮游植物为浮游动物生物量的４．５０倍）．颗粒悬浮物浓度（ＴＭ）与水体透明度（Ｓ）呈显著相关性     

对虾的行为生态学研究进展

由于对虾特殊的生态习性及其在海水养殖业中的重要地位,许多国家都开展了对虾行为生态学研究.本文综述了近年来国内外关于对虾行为习性的研究进展,着重介绍了不同生境中对虾的摄食、运动、防御、交配、产卵、蜕皮及潜底和浮现等行为习性,并探讨了不同生境中对虾行为习性的差异.在此基础上,提出了目前在对虾行为学研究中存在的一些问题以及今后的研究方向,即对虾在不同生境中的摄食策略及机制、对环境因子的选择及适应能力和对虾行为学理论在生产实践中的应用等.     

Taura syndrome,a disease important to shrimp farms in the Americas

World Journal of Microbiology and Biotechnology -     

对虾行为生态学研究进展Ⅱ.环境因子对对虾行为习性的影响

综述了近年来国内外关于外界环境因子对对虾行为习性影响的研究进展.着重介绍了光、水流和潮汐、溶解氧、底质及水温等环境因子对对虾摄食、运动、产卵、蜕皮及潜底和浮现等行为习性的影响,并探讨了对虾对环境因子变动的行为调节.提出了对虾行为生态学研究中存在的问题以及今后的研究方向,即对虾的行为习性和疾病传播间的关系、对虾的行为习性对养殖生态系统的影响和高密度工厂化养殖环境对对虾行为特征的综合影响等.     

白斑综合症病毒与对虾血淋巴细胞的体外结合实验

通过差速离心和蔗糖密度梯度离心,从感染了白斑综合症病毒(WSSV)的病虾头胸部分离了WSSV,利用地高辛对病毒蛋白进行了标记(DIG-WSSV),以体外培养的对虾血淋巴细胞为吸附基底,观察和分析了病毒与细胞间的结合现象及特性。以NBT/BCIP为酶反应显色底物观察到在细胞周围形成许多暗紫色颗粒,证实病毒与细胞间存在着稳定的结合。以OPD为酶反应显色底物分析了结合反应的特性:当DIG-WSSV维持恒定值时,随着血淋巴细胞数量的增加结合显色增强,细胞数量达到1.2104cells/孔,492nm处的吸光值达到饱和;当血淋巴细胞数量维持恒定值时,随着DIG-WSSV蛋白含量的增加显色增强,且在DIG-WSSV的蛋白浓度达到4g/孔时,492nm处的吸光值达到饱和;未标记WSSV可竞争抑制血淋巴细胞与DIG-WSSV间的结合作用。进一步的研究得出:4℃下,随着结合时间的延长显色增强,但继续延长结合时间显色反而减弱;缓冲液的渗透压对结合结果影响甚微,而酸性条件利于病毒与细胞间的结合。37℃孵育对病毒结合活性影响不大,55℃和70℃孵育可显著影响病毒的结合活性;短时间超声波处理病毒可增加病毒结合能力,长时间超声波处理可破坏病毒结合能力;有机溶剂处理同样可破坏病毒结合能力,其中尤以氯仿/甲醇的处理更为激烈;不同的去垢剂对病毒结合活性的影响结果不同:SDS和脱氧胆酸钠可以降低病毒的结合活性,而Triton X-100和NP-40可以提高病毒的结合活性。     

Virus diseases of farmed shrimp in the Western Hemisphere (the Americas): a review

Penaeid shrimp aquaculture is an important industry in the Americas, and the industry is based almost entirely on the culture of the Pacific White Shrimp, Litopenaeus vannamei. Western Hemisphere shrimp farmers in 14 countries in 2004 produced more than 200,000 metric tons of shrimp, generated more than $2 billion in revenue, and employed more than 500,000 people. Disease has had a major impact on shrimp aquaculture in the Americas since it became a significant commercial entity in the 1970s. Diseases due to viruses, rickettsial-like bacteria, true bacteria, protozoa, and fungi have emerged as major diseases of farmed shrimp in the region. Many of the bacterial, fungal and protozoan caused diseases are managed using improved culture practices, routine sanitation, and the use of chemotherapeutics. However, the virus diseases have been far more problematic to manage and they have been responsible for the most costly epizootics. Examples include the Taura syndrome pandemic that began in 1991-1992 when the disease emerged in Ecuador, and the subsequent White Spot Disease pandemic that followed its introduction to Central America from Asia in 1999. Because of their socioeconomic significance to shrimp farming, seven of the nine crustacean diseases listed by the World Animal Organization (OIE) are virus diseases of shrimp. Of the seven virus diseases of penaeid shrimp, five are native to the Americas or have become enzootic following their introduction. The shrimp virus diseases in the Americas are increasingly being managed by exclusion using a combination of biosecurity and the practice of culturing domesticated specific pathogen-free (SPF) stocks or specific pathogen-resistant (SPR) stocks. Despite the significant challenges posed by disease, the shrimp farming industry of the Americas has responded to the challenges posed by disease and it has developed methods to manage its diseases and mature into a sustainable industry.     

Nursery ground value of an endangered wetland to juvenile shrimps

Although urban development impacts wetlands around the world, until now there have been no studies of its effects on coastal wetlands in tropical regions of developed countries such as Australia. In fact the ecological value of such wetlands is poorly understood. This study provides an initial step in evaluating the ecological value of urbanised, tropical coastal wetlands by determining (a) the extent to which a remnant wetland, in a highly urbanised estuary in northern Australia, is used by juvenile commercial penaeid shrimps, and (b) the extent to which the shrimps rely on food chains based on wetland plants versus marine based food chains. Juvenile penaeids were abundant in the 11 wetland pools sampled. Catches included 5 commercial penaeids with two species, Fenneropenaeus merguiensis and Metapenaeus bennetae, comprising half the catch. Densities in the wetland pools were usually substantially higher than in adjacent estuarine habitats. Stable isotope analysis indicated that much of the nutrition of juvenile shrimps was supplied by marine primary producers (phytoplankton, epiphytic and epilithic algae, microphytobenthos, green filamentous algae) however the locally abundant saltmarsh grass Sporobolus virginicus was also a major contributor. In contrast, there was little indication of nutritional support from mangrove carbon. The lack of importance of mangrove carbon is surprising because the catches of F. merguiensis are often closely linked to the area or extent of mangroves, suggesting that aspects of mangrove systems other than the supply of mangrove carbon may determine the distribution of juvenile F. merguiensis.     

Histopathology of aflatoxicosis in the marine shrimp Penaeus stylirostris and P. vannamei

The acute and subacute toxicity of aflatoxin B₁ to the marine shrimp Penaeus stylirostris and P. vannamei (Order: Decapoda, Class: Crustacea) was investigated. Experimental shrimp were exposed to a range of concentrations of the toxin directly by intramascular injection (from 2 to 160 μg aflatoxin B₁/g body weight), or by multiple per os dosing with the feed (from 53 to 300 μg aflatoxin B₁/g feed) for up to 25 days. The histopathogenesis of aflatoxicosis in the aflatoxin-exposed animals was followed and found to be time and dose dependent in the hepatopancreas, mandibular organ, and in the hematopoietic organs. Less significant and/or inconsistent lesions were also observed in other organs and tissues, but a time-dose dependency was not noted. The principal lesions of aflatoxicosis in penaeid shrimp occur in the hepatopancreas and the mandibular organ. In the former organ, subacute and acute aflatoxicosis is expressed as necrosis of the hepatopancreatic tubule epithelium that proceeds from the proximal (older) portion of the tubules, in the center of the organ, to the peripheral (younger) tubule tips. A marked intertubular hemocytic inflammation followed by encapsulation and fibrosis of affected tubules follows in subacute aflatoxicosis, but is not as developed as in acute aflatoxicosis. The mandibular organ in aflatoxicosis displays a necrosis of the peripheral epithelial cells of the cords within the gland that progresses proximally to the central vein. Only a slight hemocytic inflammation accompanies the degenerative changes in this latter organ.