白斑综合征病毒对脊尾白虾的致病性研究

2010年、2011年江苏沿海多数地方海水养殖中发生了脊尾白虾大规模死亡的现象,本课题通过对患暴发性流行病的脊尾白虾(Exopalaemon carinicauda)进行病毒分子生物学检测,发现病虾体内白斑综合征病毒(White Spot Syndrome Virus,简称WSSV)检测普遍呈阳性;应用发病虾组织制备的病毒粗提液,进行了人工感染实验,实验表明该病毒对脊尾白虾具有较强的致病性,可引起68.5%的死亡率。电镜结果表明,发病虾与感染病虾鳃及肝胰脏等组织器官都发生了相同的细胞病理变化,主要表现为细胞核内染色质边聚、线粒体肿胀、内嵴消失等一系列细胞病理变化,在其细胞核与细胞质中发现了具有单层囊膜结构的对虾白斑综合征病毒粒子,病毒粒子大小约为(130~170)nm×(305~405)nm,核衣壳呈子弹形,一端较细另一端较粗,完整病毒粒子由核心、衣壳和囊膜构成,分子生物学实验表明该病毒为WSSV。实验结果符合柯赫氏法则,可以初步推断WSSV感染脊尾白虾并与引发的大规模死亡有着直接关系。     

Assessment of chemical and physical treatments to selectively kill non-indigenous freshwater zooplankton species

Non-indigenous zooplankton species pose a biosecurity threat to New Zealand’s freshwater native taxa. Nine species are known to have established in New Zealand lakes to date. The spread of some zooplankton taxa is linked to the translocation of farmed fish, principally grass carp (Ctenopharyngodon idella), and recreational vessel movements. The aims of this study were to assess the effectiveness of a range of chemical and physical treatments for transport water and associated equipment to kill freshwater cladoceran, copepod, and rotifer zooplankton species, and their risk to non-target fish. Sodium chloride was the most effective and applicable chemical treatment tested at length in the cladoceran and, combined with physical treatment via mechanical filtration of water or hot water immersion of equipment (to also manage the risk of diapausing eggs), represents an effective option for the control of non-indigenous zooplankton, with limited impact on stenohaline fish.     

2014年黄骅港海域浮游动物群落结构季节变化及其影响因素研究

为了解黄骅港海域浮游动物群落结构的季节变化特征及其与环境因子的相关关系,分别于2014年2月(冬季)、5月(春季)、8月(夏季)和11月(秋季)对该海域进行4个航次的浮游动物和环境因子调查,采用优势度指数、物种多样性和聚类分析等方法分析该海域浮游动物的群落结构特征及其季节变化,采用冗余分析探究引起浮游动物群落结构变动的...     

In vivo neutralization assays by monoclonal antibodies against white spot syndrome virus in crayfish(Cambarus proclarkii)

The neutralizing activities of eight monoclonal antibodies （MAbs） against white spot syndrome virus （WSSV） （2D2, 2B2,1D2, 1D5, 1C2, 4A1, 6A4 and 6B4） were analyzed by in vivo experiments. Gills from WSSV-infected shrimp were homogenized and ten-fold serially diluted by PBS, and then incubated with MAbs （hybridoma culture supernatant）, respectively. The mixture of WSSV and MAbs were injected into crayfish （Procambarus clarkii）. After challenge, the death rates of crayfish were counted to determine the neutralizing activities of MAbs. At the same time, the mixture of myeloma culture supernatant and WSSV or PBS was served as positive or negative control, respectively. The results showed that, at each virus dilution, the mean time to death of the crayfish injected with MAb-treated virus was significantly longer than that in the positive control, though they all showed 100% mortality within 25 d, and meanwhile, few crayfish died in the negative control. Among the eight MAbs, 2D2, 2B2, 1D2 and 1D5, especially the former two, delayed the mortality significantly, and 1 C2, 4A1 and 6A4 delayed the mortality as well but not so efficiently, while MAb 6IM was efficient only when the virus concentration increased. The results indicated that the anti-WSSV MAbs can neutralize WSSV in different virus dilutions.     

Changes in zooplankton community structure in Sanmen Bay, China

Seasonal variations in zooplankton abundance,biomass,species diversity and community structure were investigated in the Sanmen Bay,China.Samples were collected from 15 stations,on the seasonal basis,in April(spring),July(summer) and October 2005(autumn) and January 2006(winter),respectively.The results show that zooplankton species number,abundance and biomass varied widely and had distinct spatial heterogeneity in the Sanmen Bay.A total of 72 species of zooplankton belonging to 56 genera and 17 groups of pelagic larvae were identified.The zooplankton species richness was strongly related to salinity.Based on hierarchical cluster analysis,zooplankton in this study area were classified into three groups:coastal,neritic and pelagic groups,which corresponded to the upper,middle and lower portion of the Sanmen Bay,respectively.The coastal low-saline species were dominant in the study area.The zooplankton abundance and biomass reached a peak in summer,moderate in spring and autumn,and the lowest in winter.Zooplankton abundance decreased from the upper to lower portion of the bay in April,when the highest biomass occurred in the middle portion of the bay.There were the same spatial distribution patterns for the biomass and abundance in July,with the maximum in the middle of the bay.However,zooplankton abundance was the highest in the middle of the bay in October,when maximum biomass occurred near the lower of the bay.Zooplankton abundance and biomass were evenly distributed in the Sanmen Bay in January.Spatial and temporal variations in zooplankton and their relationship with environmental factors were also analyzed.The BIOENV results indicate that the combination of chlorophyll a(Chl-a),salinity,dissolved inorganic nitrogen(DIN),dissolved oxygen(DO) and silicate(SiO3) was responsible for the variations in zooplankton community structure in the Sanmen Bay.The environmental changes played an important role in changes in the zooplankton community structure in the Sanmen Bay.     

两种围栏养殖模式防白斑综合征病毒(WSSV)效果的比较

2003年在乳山养殖场采用两种围栏养殖模式养殖了凡纳滨对虾,并对其防白斑综合征病毒(WSSV)效果进行了比较。结果表明,凡纳滨对虾在养殖过程中WSSV阳性率有下降趋势,浮游动物WSSV阳性率较低,布围隔的防病毒效果好于网围隔。     

白斑症病毒感染与病毒囊膜完整性的关系

对白斑症病毒（white spot syndrome virus,WSSV）感染与病毒囊膜完整性关系的探讨,是深入研究切断WSSV感染途径,为该病的预防提供一条新思路、新方法并积累实验依据的重要环节.本实验利用物理低渗方法即用蒸馏水使WSSV的囊膜破碎,破坏了病毒囊膜完整性,并用此病毒做螯虾体内、外感染实验.结果发现,WSSV的囊膜破碎后仍具有感染性;在此基础上,分别用抗WSSV囊膜蛋白的单克隆抗体（4G9）和抗WSSV核衣壳蛋白单克隆抗体（2A3）进行螯虾体内中和实验,结果显示抗WSSV囊膜蛋白的单抗有中和作用,而抗WSSV核衣壳蛋白的单抗无中和作用;将破碎的囊膜从核衣壳上去掉后,用不同浓度的纯核衣壳重复以上体内感染实验,结果其感染性消失,说明囊膜破碎的WSSV引起的感染是由WSSV上的破碎囊膜介导的病毒侵染所致,而不是核衣壳被直接内吞引起的.这些结果提示,WSSV是否引起靶细胞感染主要与病毒囊膜上与病毒侵染有关的功能蛋白有直接关系,只要这些功能蛋白活性不变并且存在的量足以与靶细胞膜结合,即使WSSV囊膜破碎、结构不完整也能引起感染.     

2010年夏季雷州半岛沿岸海域浮游动物群落结构特征

2010年7月对雷州半岛海岸带的浮游动物群落结构及相关环境因子进行了调查,共鉴定出浮游动物87种和34类浮游幼体,其中原生动物4种,占总种类数的4.6%;腔肠动物20种,占总种类数的23.0%;介形类1种,占总种类数的1.1%;枝角类2种,占总种类数的2.3%;桡足类37种,占总种类数的42.5%;十足类2种,占总种类数的2.3%;毛颚动物7种,占总种类数的8.0%;被囊动物6种,占总种类数的6.9%;浮游软体动物8种,占总种类数的9.2%。优势种有13种和3类浮游幼体,孔雀强额哲水蚤和小毛猛水蚤是该海域重要的优势种群,几乎在全海域均为优势种,桡足类幼体为全海域优势类群。半岛沿岸海域浮游动物种类数由近岸往远岸逐渐增多,浮游动物丰度分布则相反。浮游动物种类数由多到少的顺序为东北部、东南部、西北部和西南部、南部,丰度由大到小的顺序为西北部、西南部、东北部、东南部、南部,生物量由大到小的顺序为东北部、西南部、西北部、东南部、南部,浮游动物丰度和生物量的平面分布反映了人类对海岸带开发利用的活动概貌。多样性指数和均匀度平均值分别为2.96和0.57。相关分析表明:浮游动物丰度与水温、细菌总数、浮游动物生物量均呈极显著的正相关关系,与磷酸盐呈显著的负相关关系;浮游动物生物量与TOC、细菌总数、叶绿素a、浮游植物丰度均呈极显著的正相关关系,与pH呈显著的正相关关系,与盐度呈显著的负相关关系。     

Enhanced resistance of marine shrimp Exopalamon carincauda Holthuis to WSSV by injecting live VP28-recombinant bacteria

Recombinant bacteria secreting the transmembrane-truncated region of white spot syndrome virus (WSSV) envelope protein VP28 named DhpVB were constructed, using live Escherichia coli as a deliverer and a constitutive secretory expression plasmid as the expression vector. With the ability to deliver recombinant protein products outside, DhpVB could make VP28 directly interact with the shrimp when injected into shrimp. In order to test the protection potential, live DhpVB were injected into the marine shrimp Exopalamon carincauda Holthuis for three times and WSSV challenge was performed twice simultaneously at the last two injections of the bacteria. The results showed that DhpVB displayed statistically significant advantage over bacteria without VP28 after the second challenge with an average RI (Resistance index) of 0.67 ± 0.08, compared with 0.41 ± 0.09 of bacteria without VP28 (P <0.05). The data suggested that a quasi-immune response may exist in E. carincauda and live bacteria carrying VP28 could enhance the shrimp resistance against WSSV.     

兔抗对虾白斑症病毒(WSSV)独特型抗体的制备及其特性分析

以前期已制备的抗WSSV囊膜蛋白单克隆抗体4G9(Ab1)杂交瘤细胞生产小鼠腹水,腹水经辛酸-硫酸铵法、ProteinG亲和层析法纯化后,用以制备兔抗血清,所得血清经纯化得到粗提的兔抗WSSV独特型抗体(Ab2)。采用竞争酶联免疫吸附实验、间接免疫荧光法、斑点免疫印迹和蛋白免疫印迹等实验方法分析了Ab2特性,结果表明:Ab2能识别Ab1并与WSSV竞争Ab1的抗原结合位点,是具有模拟WSSV特性的抗独特型抗体;Ab2能与中国对虾血细胞结合,并能部分阻断WSSV与血细胞膜的结合;其与中国对虾血细胞膜上结合的蛋白分子量分别为94.5、51.5和27.0kDa,由此推断,这三个蛋白为WSSV在中国对虾血细胞膜上的结合蛋白。本研究结果可为进一步研究WSSV侵染机理提供资料。     

White spot syndrome virus envelope protein VP124 involved in the virus infection

White spot syndrome virus （WSSV） is one of the major shrimp pathogens causing large economic losses to shrimp farming. In an attempt to identify the envelope proteins involved in the virus infection, purified WSSV virions were mixed with three antisera against WSSV envelope proteins （VP39, VP124 and VP187 ）, individually. And then they were injected intramuscularly into crayfish （Procambarus clarkii） to conduct in vivo neutralization assays. The results showed that for groups injected with virions only and groups injected with the mixture of virions and antiserum against VP124, the crayfish mortalities were 100% and 60% on the 8th day postinfection, individually. The virus infection could be delayed or neutralized by antibody against the envelope protein VP124. Quantitative PCR was used to further investigate the influence of three antisera described above on the virus infection. The results showed that the antiserum against VP124 could restrain the propagation of WSSV in crayfish. All of the results suggested that the viral envelope protein VP124 played a role in WSSV infection.     

养殖牙鲆鳗弧菌疫苗的研究

以牙鲆病原性鳗弧菌M3为抗原,制备了细胞（CV）、细胞－胞外产物（CEV）、脂多糖（LPS）三种疫苗,通过浸泡和注射两种途径对牙鲆进行两次免疫,检测疫苗的免疫保护效果。免疫后4周可检测到免疫组牙鲆具有明显的凝集抗体效价,其中注射组效价显著高于浸泡组,而对照组变化不明显。与对照组相比,免疫组牙鲆获得良好的免疫保护力,其中注射免疫组高于浸泡组。在注射免疫途径中,以CEV的免疫保护效果最好;在浸泡免疫途径中,以CEV和LPS的免疫保护效果最好。结果表明鳗弧菌M3的细胞－胞外产物苗有显著的免疫保护作用。     

2019–2020年夏季舟山海域浮游动物优势种生态位及其生态分化

为了解舟山海域浮游动物种间关系及环境因子变化对优势种生态位分化的影响,根据2019年夏季和2020年夏季舟山海域浮游动物调查资料,利用优势度指数判断该海域浮游动物优势种,运用Shannon公式和Pianka公式分析浮游动物优势种生态位宽度和生态位重叠指数,采用冗余分析方法探究影响舟山海域浮游动物优势种生态位分化的主要环境因子。结果表明,舟山海域2019年、2020年浮游动物优势种更替率为66.67%。根据舟山海域浮游动物优势种生态位宽度值,可将其分为广生态位种、中生态位种、窄生态位种,2019年广生态位种有4种,2020年有6种,分别占浮游动物优势种的66.67%和60.00%,表明广生态位种是舟山海域浮游动物优势种的主要组成成分。2019年浮游动物优势种生态位重叠指数范围为0.11～0.79,2020年生态位重叠指数范围为0～0.98。冗余分析表明,2019年浮游动物优势种生态位分化主要受硅酸盐浓度、化学需氧量、温度和叶绿素a浓度的影响,2020年浮游动物优势种生态位分化主要受硅酸盐和溶解氧浓度的影响。该研究有望进一步了解舟山海域浮游动物群落的种间关系,为阐述浮游动物对环境变化的适应...     

白斑综合症病毒膜蛋白VP124参与病毒的感染

白斑综合症病毒(white spot syndrome virus,WSSV)是对虾的最主要病毒病原之一,给对虾养殖造成了巨大的经济损失。WSSV分别与三种该病毒膜蛋白VP39,VP124和VP187的特异性抗血清在体外作用后,再注射到螯虾体内,进行中和试验。结果显示,注射同或未同VP124抗血清作用的WSSV,8 d后螯虾的死亡率分别是100%和60%,WSSV的感染能够被抗VP124抗体中和。进一步利用定量PCR分析上述三种特异性抗血清对病毒感染的影响,结果显示,VP124抗血清可抑制WSSV在螯虾体内的增殖。所有结果表明,VP124在病毒的感染中起作用。     

虾池中桡足类及其休眠卵携带对虾白斑综合症病毒的初步调查研究

探讨对虾白斑综合症(White Spot Syndrome WSS)与虾池中桡足类之间的关系,于2003年5~10月份,在暴发过对虾白斑综合症的池塘中采集桡足类样品共128个,并从冬季底泥样品分离出桡足类休眠卵样品20个,以及此种休眠卵孵化得来的桡足类幼体样品5个。利用PCR斑点杂交法以及巢式PCR法检测样品中对虾白斑综合症病毒(White Spot Syndrome Virus WSSV)携带情况。结果表明,5~10月份桡足类样品中均能检测到阳性样品,总的阳性率为56.3%;其中,5月份样品阳性率为37.5%,6,8,9,10月份各取样1次,每次样品中的阳性率均在60%以上。在20例桡足类休眠卵样品中有1例为阳性,阳性率为5%;并且实验室中孵化此种休眠卵得来的桡足类幼体样品中检测到阳性,阳性率为40%。本研究初步表明,桡足类是WSSV的携带者或传播媒介之一,且冬季池塘底泥中携带WSSV的桡足类休眠卵可萌发出带病毒的桡足类。     

广东省南澳岛东部海域浮游动物群落结构及其影响因素

广东省南澳海域是粤东重要的海产养殖基地, 分析该海域浮游动物群落结构特征对评估其生态环境质量具有重要意义。文章根据2014年9月(秋季)、12月(冬季)、2015年4月(春季)和2016年7月(夏季)在南澳岛东部海域的浮游动物调查, 分析该海域浮游动物的群落结构特征, 探讨环境因素对其时空分布的影响。共鉴定浮游动物206种(包括浮游幼虫), 桡足类种数最多, 达94种; 远岸海域浮游动物的种数高于近岸海域。浮游动物丰度和生物量的季节变化明显, 夏、秋季高于冬、春季; 浮游动物丰度和生物量的分布趋势较一致, 夏季高值区主要出现在近岸, 秋季由近岸向远岸海域递增。浮游动物不同类群和优势种的丰度也存在季节变化, 桡足类是调查期间丰度较高的类群, 秋季水母类和海樽类丰度明显增加; 优势种后圆真浮萤(Euconchoecia maimai)和针刺真浮萤(Euconchoecia aculeata)在夏季丰度高, 小齿海樽(Doliolum denticulatum)在秋季占绝对优势。温度、盐度和浮游植物生物量是影响南澳岛东部海域浮游动物时空变化的主要环境因子, 说明该海域浮游动物群落特征受海流、水团和养殖活动的综合影响。     

春季我国不同纬度河口浮游动物群落变化趋势

根据2010-2012年春季由南向北河口海域:北仑河口、九龙江口、瓯江口、长江口和灌河口的浮游动物调查资料;通过比较不同纬度河口浮游动物群落的生物多样性指数（H'）、物种更替率（R）和群落相似系数（S）;探讨了河口浮游动物群落特征沿着不同纬度变化的趋势。结果表明;自南向北5个河口可以划分为3个不同的浮游动物区系。北仑河口和九龙江口为南部亚热带区系;位于亚热带海域;以亚热带种为主;H'分别为1.01和2.18;两河口之间R值为65.52%;S值为51.28%;瓯江口和长江口为中部过渡区系;位于亚热带和温带过渡海域;亚热带种和暖温带种是主要出现的种类;H'分别为1.31和1.86;两河口之间R值为58.33%;S值为58.82%;灌河口单独为北部温带区系;位于温带海域;暖温带种是主要出现的种类;H'值为1.19;与其余河口物种更替率均高于80.00%;群落相似系数均低于32.00%;表明群落结构由南到北的变化非常显著。线性回归结果表明:我国不同河口之间浮游动物的物种更替率与纬度差（两河口所处纬度的差值;δN）呈正相关关系（R=0.026ΔN+0.608;p=0.002）;群落相似系数与纬度差呈负相关关系（S=0.034ΔN+0.578;p=0.001）。春季;我国不同纬度河口浮游动物群落结构的变化;主要是因为影响各河口的水温和水团存在较大差别所致。     

荣成湾毗邻海域浮游动物群落季节分布特征

利用2009年度4个季度月的现场调查资料研究了荣成湾毗邻海域浮游动物群落的季节分布特征。研究表明, 在荣成湾毗邻海域出现的浮游动物共59个种类, 秋季、夏季和春季浮游动物均为30种以上(依次为38种、35种和32种), 而冬季种类数相对较少(22种)。浮游动物优势种主要为强壮箭虫(Aidanosagitta crassa)、中华哲水蚤(Calanus sinicus)、小拟哲水蚤(Paracalanus parvus)和拟长腹剑水蚤(Oithona similis)等种类, 其个体丰度的空间和季节分布特征差异较大。浮游动物湿重生物量冬季最高(433.8 mg/m3), 春季(220.7 mg/m3)和秋季(162.4 mg/m3)次之, 而夏季(52.5 mg/m3)相对较低。浮游动物群落物种多样性指数H′年平均值为2.50, 均匀性指数J′年平均值为0.65, 季节比较上, 夏季浮游动物多样性和均匀度均较高, 而春季多样性和均匀性平均值为低值, 且最大值和最小值之间波动范围较大。进一步分析显示, 湾内的筏架养殖对浮游动物群落的分布有显著影响。     

2011—2020年调水调沙前后黄河口海域浮游动物年间变化及影响因子

依据2011—2020年(2016年中断)黄河调水调沙工程实施前、后黄河口海域18个站位野外调查数据, 研究浮游动物种类组成和丰度变化及优势种更替, 以查明黄河口海域浮游动物的年间变动规律及调水调沙工程的影响。结果显示: 共记录浮游动物82种类(包括浮游幼体16类), 其中刺胞动物种类数最多, 共记录26种, 其次是桡足类出现23种。浮游动物种类组成的年间变化差异显著(P < 0.05), 2017年(未进行调水调沙)最低; 调水调沙前、后分别记录浮游动物69种和75种, 调水调沙前后种类数呈正相关(r=0.684, P=0.042); 调水调沙前后优势种更替率较高; 浮游动物丰度在2011—2015年较高, 2017—2020年相对较低, 调水调沙后浮游动物丰度年间变化达极显著水平(P < 0.01), 而调水调沙前后没有显著差异; 浮游动物丰度水平分布总体呈现从黄河口近岸到离岸递减趋势, 密集区位于黄河口北部近岸海域, 调水调沙后密集中心较调水调沙前向离岸海域略有推移。冗余分析表明, 对黄河口浮游动物年间变化影响较大的因子是叶绿素a浓度、溶解氧浓度和水温。研究表明: 2011—2020年调水调沙期间黄河口海域浮游动物丰度年间变动大于调水调沙前后变动, 调水调沙实施对浮游动物影响持续时间较短。     

日照岚山港北作业区邻近海域春、秋季浮游动物群落结构研究

以2015年5月(春季)和10月(秋季)在日照岚山港北作业区邻近海域进行的浮游动物调查数据为例,分析了该海域浮游动物的群落结构特征.调查海域两季共发现浮游动物成体20种,浮游幼虫7类,其中春季成体12种,浮游幼虫4类,秋季成体19种,浮游幼虫6类;春、秋两季调查浮游动物平均丰度分别为128.1 ind./m3、103....