扇贝养殖海区中小型浮游生物携带AVNV的荧光定量检测

为了解中小型浮游生物与扇贝急性病毒性坏死症(AVND)的流行传播关系,于2009年对青岛流清河与荣成桑沟湾2个不同扇贝养殖海区的浮游生物进行了采样调查,利用荧光定量PCR(Fluorescent quantitative PCR,FQ-PCR)技术对所采集样品携带急性病毒性坏死病毒(AVNV)的情况进行了定量检测.结果表明,2个扇贝养殖海区的浮游生物样品均检测到AVNV,而且不同粒径的浮游生物携带AVNV载量有差异(P<0.05):小于200目粒径的浮游生物携带AVNV载量最大,达到9.45×106 copy/mg (DNA); 60～200目粒径的浮游生物次之,大于60目粒径的浮游生物携带量最少,仅为5.81×104 copy/mg(DNA).在上述2个扇贝养殖海区,均在8月份检测到浮游生物携带AVNV.该结果与栉孔扇贝夏季大规模死亡在时间上是吻合的.结论认为,中小型浮游生物可以携带AVNV,并有可能在扇贝急性病毒性坏死症的流行和传播中起到传播媒介的作用.     

间接免疫荧光法检测栉孔扇贝急性病毒性坏死症病毒

用纯化栉孔扇贝(Chlamys farrei)急性病毒性坏死症病毒(AVNV)免疫兔子,以兔抗血清为一抗,荧光标记的羊抗兔抗体为二抗,采用冰冻切片技术,建立了栉孔扇贝AVNV的间接免疫荧光检测方法。用该方法分析栉孔扇贝和海湾扇贝(Argopecten irradians)体内AVNV感染强度,并对感染率进行统计。结果表明,栉孔扇贝的肾脏、肝胰腺中,AVNV阳性信号最强,呈现中度到重度感染,其AVNV感染率100％。鳃丝、性腺及闭壳肌中未检测到阳性信号。在海湾扇贝的肝胰腺及肾中AVNV的感染率也较高。提示AVNV感染扇贝的靶器官主要是肾脏、肝胰腺。对与栉孔扇贝同一海区养殖的海湾扇贝在栉孔扇贝发病期间存活率较高的原因进行了讨论。     

栉孔扇贝病原感染与病害发生关系探讨

利用电镜负染技术检测自然海区养殖和人工感染试验的栉孔扇贝组织提取液,分别发现病毒和类立克次氏体(RLO)两种病原体。动态检测两类病原体感染状况与整个养殖期间栉孔扇贝发病死亡情况,结果表明,养殖期间病毒感染率在80％—100％之间,最大感染强度出现在栉孔扇贝大规模死亡的7、8月份。人工感染病毒的栉孔扇贝,其病毒感染率和感染强度与养殖栉孔扇贝大规模死亡时期相当。而两种情况下栉孔扇贝类立克次氏体的感染率和感染强度均未见明显变化。综合分析表明,病毒可能是造成栉孔扇贝大规模死亡的直接病原体。     

应用DGGE技术研究扇贝养殖海域微型真核浮游生物多样性

为了研究扇贝养殖海区微型真核浮游生物群落多样性,明确养殖扇贝发病时期高丰度微型真核浮游生物种类,探讨微型真核浮游生物与栉孔扇贝急性病毒性坏死病毒(acute viral necrosis virus,AVNV)水平传播的可能关系。于2009年和2010年从青岛流清河湾扇贝养殖海区采集了9个月份的海水样品,经25和3 μm的滤膜过滤收集海水中3~25 μm的浮游生物,扩增18S rDNA可变区序列,并利用变性梯度凝胶电泳(denaturing gradient gelelectrophoresis,DGGE)技术对扩增序列进行分离以分析微型真核浮游生物多样性。结果表明,该养殖海区微型真核生物包括甲藻、纤毛虫、眼虫、定鞭藻、硅藻、盘蜷虫、隐藻、领鞭毛虫、变形虫和Cercozoan,其中甲藻类和纤毛类生物的最高相对丰度分别达41.0%和38.2%,是海区的优势种类。各月份DGGE谱带聚类分析结果表明,2009年6、7、8、9月份浮游生物群落组成较为相似。中肋骨条藻在扇贝发病前后均有分布。结合相关扇贝AVNV已有的研究结果,研究认为中肋骨条藻是AVNV水平传播的参与者之一,但海区中广泛分布的甲藻和纤毛虫与AVNV传播的关系还有必要进一步研究。     

烟台四十里湾栉孔扇贝、海带和刺参负荷力的模拟测定

利用模拟养殖系统,实测了四十里湾海区对栉孔扇贝（Ｃｈｌａｍｙｓ ｆａｒｒｅｉ）、海带（Ｌａｍｉｎａｒｉａ ｊａｐｏｎｉｃａ）和刺参（Ａｐｏｓｔｉｃｈｏｐｕｓ ｊａｐｏｎｉｃｕｓ）的负荷力。结果表明,模拟养殖系统,单养和贝藻参混养系统对栉孔扇贝的负荷力分别为１８４．６和２０１．７ｇ／ｍ＾２;混养系统对海带和刺参的负荷力分别为１０６．８和７４７．７ｇ／ｍ＾２。该海区对单养和贝藻参混养系统栉孔扇贝的负荷     

套子湾刺参半人工采苗技术

野生稚参比同期人工培育的稚参体质健壮、肉质肥厚、生长速度快,深受养殖业户的欢迎。养殖栉孔扇贝和牡蛎都能不同程度地采集到一定数量的稚参。受此启发,去年在总结往年生产经验的基础上,我们在套子湾扇贝养殖海区结合栉孔扇贝的养殖和栉孔扇贝半人工采苗生产投挂刺参采苗器,进行了刺参半人工采苗试验,现报告如下。     

桑沟湾贝藻养殖区附着生物群落季节演替研究

从2007年5月～2008年4月,采用悬挂试网和直接从海区采集栉孔扇贝养殖笼样品两种方法,对桑沟湾贝藻养殖区的附着生物群落结构和季节演替规律进行了研究。结果显示,桑沟湾附着生物群落结构复杂,本实验鉴定的大型附着生物23种,玻璃海鞘、柄海鞘和紫贻贝是群落中的优势种。附着生物的生物量呈现明显的季节变化特征,试网上附着生物湿重与温度变化相关,生物量为0.003～1.21kg/m2,其中2月和8月分别具有最小和最大值。栉孔扇贝养殖笼的附着生物生物量9月为1.94kg/笼,之后随温度下降而减少,10月为0.99kg/笼,之后又有所上升,11月份为1.03kg/笼。生物量变化主要是由于优势种的演替引起的,随温度下降海鞘类逐渐消退,贻贝等成为优势种。     

扇贝养殖海区浮游生物携带AVNV的荧光定量分析

为了探明养殖海区浮游生物与AVNV流行传播的关系,实验自2009年5月至11月,分别从青岛沙子口和荣成桑沟湾两个扇贝养殖海区定期采取50和300 cm两个水层水样,经25、3和0.22 μm 3个孔径的滤膜分级过滤收集3个粒径的浮游生物组分,利用荧光定量PCR技术对浮游生物携带AVNV的情况进行了定量检测。结果表明两个海区浮游生物样品均检测到AVNV,检测的AVNV数量也均在8月份达到峰值,沙子口海区每升海水中浮游生物携带4.11×10⁶拷贝的AVNV,桑沟湾海区浮游生物携带1.49×10⁵拷贝的AVNV。两个海区3个粒径浮游生物组分携带AVNV的数量从高到低依次为0.22~3 μm组分、3~ 25 μm组分和大于25 μm组分,但两个水层(50和300 cm)浮游生物携带AVNV的数量则没有显著差异。结合养殖期间扇贝摄食浮游生物的特点,研究认为,扇贝在养殖期间可能会通过滤食携带病毒的浮游饵料而感染AVNV,海区浮游生物可能是导致AVNV流行的重要传播媒介。     

栉孔扇贝换季养殖方法

近几年来，山东省日照市东港区浅海筏式养殖的栉孔扇贝连续几年出现大面积的死亡现象，死亡率高达９０％，部分海区甚至绝产。针对这种情况，我们及时进行了养殖结构的调整，推广栉孔扇贝的换季养殖，就是改变“春放秋收”的传统养殖方式为“秋放春收”，避开夏季高温死亡期和台风期，有效地制止了浅海养殖栉孔扇贝的滑坡现象，取得了较好的经济效益。现把我区林孔扇贝换季养殖的做法介绍如下： １、海区及器材养殖海区和养殖器材均与原养殖方式相同。 ２、苗种栉孔扇贝换季养殖的关键因素之一是购得大规格的苗种。大规格扇贝苗来源有二种，…     

招远县大力发展扇贝养殖

招远县濒临莱州湾，浅海可养面积10万亩，水质肥沃，饵料生物繁多，适宜发展海水养殖。1984年11月从长岛引进栉孔扇贝苗种，1985年7月从乳山县引进海湾扇贝苗种进行试养，结果栉孔扇贝经11个月养殖，壳高63毫米，亩产3．493公斤；海湾扇贝经4个月养殖，壳高47毫米，亩产2．260公斤。     

贝壳上污损生物的重量对栉孔扇贝生长和存活的影响

对网笼养殖的栉孔扇贝壳上污损生物的数量进行了研究,发现9～11月贝壳上污损生物的湿重分别为1．47、0．49和2．09g,与上壳重的比值分别为28．16％、10．24％和31．29％。通过在栉孔扇贝上壳添加上壳干重0．5、1、2和3倍重的水泥,用以模拟附着在扇贝壳上的污损生物,设不添加水泥的对照组,对贝壳上污损生物的重量对栉孔扇贝生长和存活的影响进行了研究。结果表明,实验组扇贝壳长、闭壳肌和剩余软体组织的特定增长率及存活率与对照组均没有显著差异（P〉0．05）,说明贝壳上的污损生物的重量没有影响扇贝的生长和存活。     

Pathogenicity and infectivity of a rickettsia-like organism in the sea scallop, Placopecten magellanicus

Abstract. A rickettsia-like organism from the gills and plicate membrane of the sea scallop, Placopecten magellanicus , was prevalent in near-shore Rhode Island scallop populations such as Block Island Sound and Narragansett Bay, but relatively scarce in Georges Bank samples. Fifty-three per cent of scallops transplanted in cages to Narragansett Bay became infected with the rickettsiae in 2.5-3.5 months. Scallops that were transferred to aquaria with Narragansett Bay water were 100% infected with rickettsiae in 5 months or less. There is some evidence that heavy infection of P. magellanicus inhibits swimming response and that rickettsial infection may contribute indirectly to mortalities by reducing scallop mobility and avoidance of predators. The organism was successfully cultured in gill organ culture and by direct injection of infected tissue homogenate, but could not be grown on artificial media or in yolk sac of embryonated hens' eggs. Other bivalve species could not be infected with the rickettsia of P. magellanicus through the water column or by direct injection.     

The effects of epibionts and predators on the growth and mortality rates of Argopecten purpuratus cultures in southern Chile

Epibiont and predator effects on the growth and mortality of the northern scallop Argopecten purpuratus were evaluated in cultures in southern Chile.The most common epibionts were bryozoans, hydrozoans and algae in both study sites, Metri Bay (41°36; 72°43W) and Chidhuapi Channel (41°49S; 73°05W).After 12 months of culture in pearl nets and lantern nets, the scallops size did not show statistically significant differences in cultures with and without epibionts in both study sites nor were there differences in the growth either with or without the presence of predators (decapod crustaceans). The growth rate was higher in Chidhuapi Channel than in Metri Bay.Mortality was concentrated in the initial phase of culture in pearl nets. During the culture phase in lantern nets, the mortality rate was lower than 3%. The mortality rate in scallops with epibionts was higher than when these were removed in the culture phase in pearl nets. In Metri Bay the mortality rate with predators was higher than without predators. Epibionts and predators did not affect mortality in the culture phase in lantern nets.Epibionts and predation are important factors in the early mortality of scallops and therefore in the success of culture. Epibiosis, however, is not important in scallop growth in southern Chile. This is related to the composition of the epibionts and to the low temperatures which probably limit the growth of algae and invertebrates.     

栉孔扇贝急性病毒性坏死病毒（AVNV）卵内的垂直传播途径

急性病毒性坏死病毒已被证实为是一种对养殖栉孔扇贝危害极大的病原体,探讨其传播途径是防控扇贝“大规模死亡症”的前提之一。将亲贝各放在一独立育苗水体内产卵、孵化,并进行幼虫培育。通过春季和秋季室内人工育苗,从40个独立育苗水体中,采获亲贝、卵各23个家系样品,受精卵、D形幼虫、壳顶幼虫各12个家系样品,眼点幼虫2个家系样品。采用间接ELISA法、间接免疫荧光技术和电子显微镜技术对亲贝、卵和发育各期的幼虫进行定性和定位的检测。结果发现,栉孔扇贝的性腺组织中存在病毒粒子,而卵、受精卵及各期发育幼虫体内均未检测到病毒。由此可以判断：AVNV可以感染栉孔扇贝的性腺,但并不存在卵内垂直传播的可能性。     

流清河湾扇贝养殖区夏季附着生物特征研究

2007年6~9月,利用悬挂不同网目的试网方法,对夏季青岛流清河湾栉孔扇贝筏式养殖区附着生物的种类、数量及其垂直分布特征进行了研究。结果显示,流清河湾附着生物群落的优势种在6月份为麦秆虫和钩虾,7月份以后贻贝成为优势种。附着生物湿重随挂网时间延长而增加,8月份增长最快,特定增长率最高可达8.77%/d,主要是由于优势种贻贝的快速生长。玻璃海鞘等海鞘类在流清河湾较少出现。附着生物的数量受网目和水深的影响,总体上随水深的增加而下降,较小网目上附着生物量较大。贻贝为群落优势种是流清河湾夏季附着生物数量较高的主要原因。     

栉孔扇贝夏季大规模死亡的神经网络预测模型

根据2002年和2003年对山东荣成桑沟湾栉孔扇贝养殖海区的水温、盐度、pH、氨氮浓度、亚硝氮浓度等环境因子和扇贝血清中的蛋白浓度、酸性磷酸酶活力、碱性磷酸酶活力、超氧化物歧化酶活力和过氧化氢酶活力等免疫学指标及栉孔扇贝养殖密度和死亡率的监测数据,运用人工神经网络(artificial neurd network,ANN)的原理和误差反相传播(back propagefion,BP)网络的方法,利用MATLAB软件初步建立养殖栉孔扇贝夏季大规模死亡的BP人工神经网络预测模型.预测模型经过300次的学习训练,误差平方和由67.46下降至0.009 1.该预测模型对未参与模型构建的样本预测的结果与实际监测结果的符合率达到87.5%.首次将人工神经网络与水产动物病害死亡的预测相结合,建立的预测模型具有对数据适应能力强,可适时学习,预测结果准确等突出优点,为水产养殖动物病害死亡程度的预测提供了一个新的研究方法.     

Growth,survival and immune activity of scallops,Chlamys farreri Jones et Preston,compared between suspended and bottom culture in Haizhou Bay,China

We examined the growth, survival and immune response of the scallop, Chlamys farreri, during a 1‐year period in deep water of Haizhou Bay. Scallops were cultured using two methods: (1) in lantern nets at a 5 m depth and (2) in a bottom culture system (sleeves) on the seabed at about a 25 m depth. Shell heights, meat dry weight and immune activities in the haemolymph (superoxide dismutase and myeloperoxidase) were measured bimonthly or quarterly from July 2007 to June 2008. Survival was measured at the end of the study and environmental parameters in the experimental layers were monitored during the experiment. The growth and immune activities of scallops were lower when the water temperature was high, which was consistent with the main mortality occurring in summer. The growth and immunity of scallops were higher in the suspended culture than in the bottom culture during the experiment, with the exception of shell growth during the last study period. Survival of scallops in the suspended culture (54.6±12.3%) was significantly lower than that in the bottom culture (86.8±3.5%) at the end of this study. We conclude from our results that the high mortality of C. farreri can be prevented by culturing them in a bottom culture system before November of the first year, and then transferring them to a suspended culture to improve scallop production.