共查询到20条相似文献，搜索用时 328 毫秒
1.
辛志明 《中国海洋大学学报(自然科学版)》2011,41(10):40-44
本研究用纳米胶体金颗粒标记抗迟缓爱德华氏菌(AL60306NA1)单克隆抗体3A7并制备金标垫,将抗迟缓爱德华氏菌(AL60306NA1)单克隆抗体4F11与羊抗鼠抗体作为捕获抗体包被硝酸纤维素膜,建立迟缓爱德华氏菌的胶体金免疫层析快速检测方法。经过对试纸条的特异性和灵敏度测定,结果表明:试纸条与嗜水气单胞菌、温和气单胞菌、豚鼠气单胞菌、海安气单胞菌、产碱假单胞菌、无乳链球菌、大肠埃希杆菌、哈维氏弧菌、鳗弧菌、创伤弧菌、副溶血弧菌、类志贺邻单胞菌、鲁氏不动杆菌等15种水产常见病原菌没有交叉反应,与迟缓爱德华氏菌特异性反应,检测灵敏度为1×105cfu.mL-1,检测所需时间低于20min。所制备的迟缓爱德华氏菌胶体金免疫层析检测试纸条具有快速、简便、特异性高和适应基层生产推广应用等优点。 相似文献
2.
《海洋湖沼通报》2015,(3)
为明确江苏连云港养殖长牡蛎大规模死亡病因,对分离自长牡蛎的4株病原菌开展了致病性、表型特征、分子特征等研究,结果表明4株病原菌均具致病性,4株病原菌ML1、ML2、ML3、ML4对长牡蛎的半数致死量LD50分别为2.8×105、1.1×105、1.8×105和1.6×105 CFU/mL;4株病原菌均为呈短杆状革兰氏阴性细菌,氧化酶阳性,还原硝酸盐,能利用甘露醇、纤维二糖,形态及理化特性结果表明4株分离菌与弧菌属的哈维氏弧菌亲缘关系最近;16SrRNA、gyrB、rpoA基因同源性检索及系统发育学分析结果表明4株分离菌与GenBank数据库中哈维氏弧菌的基因序列相似性最高且在系统发育树中聚为1个分支;根据4株分离菌的致病性、表型与分子特征,表明引起长牡蛎大规模死亡的病原为哈维氏弧菌。为进一步明确分离菌株毒力基因的携带情况,检测了分离鉴定的4株病原菌的9种毒力基因,结果表明,4株病原菌均可检测到luxR、toxR、vhhA和vhhB 4种毒力基因,扩增片段大小分别为679bp、390bp、1324bp和216bp,其他5种毒力基因未检测到,且分离鉴定的4株病原哈维氏弧菌携带相同的毒力基因,这些毒力基因可作为检测致病性哈维氏弧菌的分子标记。 相似文献
3.
副溶血弧菌(Vibrio parahaemolyticus)是海水养殖动物弧菌病的主要病原菌之一。为实现对该菌的现场快速检测,本文以制备的兔抗副溶血弧菌多克隆抗体作为金标抗体,提取副溶血弧菌的外膜蛋白、鞭毛蛋白、胞外产物和全菌蛋白,以不同浓度的4种抗原蛋白作为4条检测线(T1~T4),特别在T4线设置全菌蛋白,基于竞争免疫层析原理研制出副溶血弧菌胶体金快速检测试纸。使用该试纸检测了副溶血弧菌(V.parahaemolyticus)、鳗弧菌(V.anguillarum)、溶藻弧菌(V.alginolyticus)、哈维氏弧菌(V.harveyi)和鱼肠道弧菌(V.ichthyoenteri),检测结果表明其能够准确鉴别副溶血弧菌感染,排除其他菌的交叉反应干扰。副溶血弧菌快速检测试纸的最低检测限为5×105 cfu·mL^-1,感染副溶血弧菌的牙鲆组织的检测结果与ELISA结果一致。本文研制的试纸具有较高的特异性与准确性且操作简单,为现场快速检测副溶血弧菌提供了有力工具。 相似文献
4.
为筛选具有广谱抑菌活性的乳酸菌用于微生态制剂开发,本研究自野生许氏平鲉肠道内共分离出35株细菌,从中筛选出1株乳酸菌YH1,其革兰氏染色呈阳性,且可发酵脱脂乳产生凝固、无气泡、呈酸性现象,基于形态学观察、生理生化和16S rDNA序列鉴定乳酸菌YH1为食窦魏斯氏菌。分析了酸碱度、温度和NaCl含量对食窦魏斯氏菌YH1生长的影响;选取金黄色葡萄球菌、嗜水气单胞菌、溶藻弧菌、副溶血弧菌、迟钝爱德华氏菌、鳗弧菌、假交替单胞菌和哈维氏弧菌等多种病原指示菌,以琼脂扩散法测定YH1培养12h产物的抑菌活性;并采用肌肉注射和腹腔注射两种方式对YH1的动物安全性进行检测。实验结果显示,食窦魏斯氏菌YH1适宜的生长条件为pH5.7—8.8、温度20—40oC,最适生长条件为pH7.2—8.4,温度35oC,在MRS培养基中添加NaCl会抑制YH1的生长;食窦魏斯氏菌YH1的无菌培养产物可显著抑制金黄色葡萄球菌、嗜水气单胞菌、溶藻弧菌、副溶血弧菌、迟钝爱德华氏菌、鳗弧菌、假交替单胞菌、哈维氏弧菌等病原指示菌的生长;食窦魏斯氏菌YH1在10~8CFU/m L及以下浓度对许氏平鲉是相对安全的。以上研究表明了食窦魏斯氏菌YH1具有广谱抑菌性和动物安全性,为其应用于水产养殖业奠定了理论基础。 相似文献
5.
采用体外拮抗试验的方法，先从不同生长阶段的对虾肠道中分离的96株细菌中筛选出4株对病原菌（鳗弧菌、副溶血弧菌、哈维氏弧菌）有拮抗作用的细菌；进一步通过固体扩散法和液体混合法确定LV060806、LV060808的产物对鳗弧菌有明显抑制作用，Lv060806、LV060815的产物对副溶血弧菌有明显抑制作用，LV060810、LV060815的产物对哈维氏弧菌有明显抑制作用。溶血试验证实这些菌株不产生溶血素，不具有潜在的致病性；动物安全性实验证实菌株浓度在10^6 cfu／ml以下对实验对虾无明显的毒害作用。随后测定了其16S rRNA基因序列，分析了相关细菌相应序列的同源性，构建了系统发生树，结合菌株表型特征和生理生化特性最终鉴定结果为：LV060806、LV060808为Vibrio natriegens；LV060810、LV060815为V.alginolyticus。 相似文献
6.
利用十二烷基磺酸钠抽提并结合超速离心的方法提取了鳗弧菌(Vibrio anguillarum)、河口弧菌(V.aestuarianus)、霍乱弧菌(V.cholerae)和副溶血弧菌(V.parahaemolyticus)4种致病性弧菌的主要外膜蛋白,通过SDS-PAGE分析比较4种弧菌主要外膜蛋白的组分。结果表明:4株弧菌的外膜蛋白电泳图谱一般可得到5～10条蛋白带,分子量多数集中在26~40 kD和48~85 kD。对所提取的4种弧菌主要外膜蛋白进行SDS-PAGE后,分别与自制的兔抗鳗弧菌血清、抗河口弧菌血清、抗副溶血弧菌血清进行Western-blotting印迹分析。结果显示每种全菌抗血清可与相应菌的外膜蛋白部分组分发生免疫反应,并与其他3种弧菌外膜蛋白的部分组分产生交叉免疫反应,这些反应条带分子量主要集中于30~48 kD之间。本研究为进一步研究致病性弧菌外膜蛋白免疫学特性提供参考。 相似文献
7.
6种海洋致病性弧菌36kDa外膜蛋白特性分析 总被引：3，自引：0，他引：3
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. 相似文献
8.
养殖石斑鱼溃疡病病原哈维氏弧菌胞外产物的研究 总被引：2，自引：0，他引：2
以网箱养殖的患溃疡病青石斑鱼(Epinephlus awoara)和斜带石斑鱼(Epinephlus co-ioides)分离的哈维氏弧菌(Vibrio harveyi)TS-628菌株为研究对象,采用平板玻璃纸覆盖技术,获得该菌株的胞外产物(ECP),并对其成分进行分析。利用杯碟法研究表明,哈维氏弧菌TS-628菌株的胞外产物具有酪蛋白酶、明胶蛋白酶、淀粉酶、脂肪酶、卵磷脂酶等多种酶活性和溶血活性,且溶血性物质具有热不稳定性。SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)及考马斯亮蓝R-250染色分析表明,ECP蛋白条带主要集中在分子质量范围25.0~66.2 ku之间,经不同时间培养所获得的ECP在凝胶上显示出的蛋白条带几乎一致。采用复性电泳方法(G-PAGE)分析了哈维氏弧菌及其它4种弧菌属细菌的ECP蛋白酶活性,结果发现哈维氏弧菌ECP在酸性条件下,蛋白酶活性较强,在碱性条件下,蛋白酶带明显减少,活性很弱。在相同的pH条件下,5种弧菌中以副溶血弧菌的ECP蛋白酶活性最强,种类多,分子质量范围宽;而哈维氏弧菌的蛋白酶带较少,活性也稍弱。 相似文献
9.
从患病黑鲪分离病原菌HV0811的鉴定及其系统发育分析 总被引：1，自引：0，他引：1
从症状为溃疡瞎眼的养殖黑鲪(Sebastodes fuscescens)病灶处分离到3株优势菌HV0811、PT0811和JH0811,以肌肉注射人工感染实验证实菌株HV0811为致病菌,其对黑鲪的半致死浓度为7.15×10~5 CFU/尾。通过对致病菌的形态学观察、生理生化分析表明:菌株HV0811为革兰氏阴性菌,极生单鞭毛,短杆状,直径1～3mm,需Na+生长,低于4℃、高于42℃不生长。基于16SrRNA和HSP60基因序列构建的系统发育树表明:HV0811分别与哈维氏弧菌(AY750578和AY332571)相类聚,其中16SrRNA基因序列与哈维氏弧菌(AY750578)聚合置信度较低,仅有10%,不能有效地确定该种;HSP60基因序列与哈维氏弧菌(AY332571)聚合为一个分支的置信度达100%。综合该菌的形态、生理生化及HSP60基因序列比对结果,表明分离到的病原菌HV0811为哈维氏弧菌。药敏试验结果显示病原哈维氏弧菌(HV0811)对庆大霉素、新霉素、氟哌酸等较为敏感。 相似文献
10.
采用凝胶层析技术从泥蚶血细胞裂解物中纯化得到两种血红蛋白Tg-HbⅡ(异源四聚体)和TgHbⅠ(同源二聚体),回收率分别为48.15%和24.91%。胰蛋白酶酶解Tg-HbⅡ,酶解液经反相C-18色谱柱分离纯化得7种血红蛋白源多肽F1~F7。通过琼脂扩散法检测酶解多肽抑菌活性,结果发现7种多肽对金黄色葡萄球菌、大肠杆菌、恶臭假单胞菌、枯草芽孢杆菌和坚强芽孢杆菌均无抑菌作用,但多肽F2~F7对副溶血弧菌、溶藻弧菌和哈维氏弧菌具有明显抗菌活性,且多肽F2~F7对副溶血弧菌、溶藻弧菌和哈维氏弧菌的MIC值在0.012~0.200mg/mL范围内。荧光显微镜观察酶解多肽杀菌效果发现多肽杀菌迅速,5min内弧菌细胞几乎全部死亡。利用透射电子显微镜观察正常的和经抗菌肽处理后的溶藻弧菌和哈维氏弧菌细胞的超微结构,结果显示,对照组正常细胞内部结构完整,而经抗菌肽处理过的细胞其细胞壁完整,但因缺乏细胞内容物支持导致局部区域内陷,细胞质膜明显收缩,细胞内物质外流形成许多空泡从而导致细胞死亡。研究结果表明血红蛋白源抗菌肽对泥蚶致病菌弧菌有明显的抗菌活性,其通过破坏细胞内部结构而起到杀菌作用,这些发现为血红蛋白抗菌机制的研究奠定了基础。 相似文献
11.
Arne Schnack-Friedrichsen Angela M. Davis Jim D. Bennell Dei G. Huws 《Marine Georesources & Geotechnology》2001,19(4):221-243
The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths. 相似文献
12.
Shigehisa Nakamura 《Marine Geodesy》2000,23(1):55-61
Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone. 相似文献
13.
The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics. 相似文献
14.
15.
《海洋学报(英文版)》2014,(9):F0003-F0003
<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology, 相似文献
16.
The relationship between the flux of exotic benthic foraminiferal tests (i.e. tests which are supplied from open-sea sources alone) in a tidal inlet and that of bulk sediment was analysed, which can be expressed as two first-order linear equations. According to this relationship, in order to determine net sediment transport directions in the entrance, the test concentration in surficial sediments of the tidal basin can be compared against a ‘ critical level ’. The critical level is determined for the conditions that no net transport of bulk sediment is present within the entrance. If the observed concentration (averaged over the tidal basin) is higher than the simulated critical level, then the net sediment transport is directed to landward. This method is applied to the analysis of net sand transport at Christchurch Harbour, a tidal inlet system located in southern England. In this investigation, concentrations of exotic foraminiferal tests in the surficial sediments of the tidal basin and ebb tidal delta area were obtained from the analysis of sea-bed sediment samples. A series of probable critical levels were calculated based upon the data sets with regard to: (1) sediment discharge from the rivers; (2) magnitude of sediment discharge within the entrance during the ebb; (3) the test concentration outside the harbour; (4) the thickness of the moving layer; and (5) two parameters associated with dispersive processes. The results show that the concentration in the tidal basin sediment is higher than a number of simulated critical concentrations for representative cases. Consequently, the high level of the concentration of exotic benthic foraminiferal tests within the harbour should be explained as a result of landward net transport of sands within the entrance. 相似文献
17.
18.
Rahul Sharma 《Marine Georesources & Geotechnology》2000,18(3):237-250
Observations on seafloor features from subbottom profiling and seabed photography data show that the undisturbed and natural conditions before the benthic disturbance undergo a substantial change after operation of the disturber. The effects can be seen in the form of deep trenches and grooves, formed by the action of the sleds and the pumping of the sediment along its path, as well as sediment piles on either side of the track. Biological traces such as fecal coils, casts, tubes, burrows, and trails get obliterated in the areas of disturbance and resedimentation. The smooth, uniform nature of the seafloor observed in predisturbance phase shows microtopographic changes in and around the disturbed area as a result of sediment excavation and resettlement. Vertical mixing as well as lateral transport of sediment alter the geological, biological, physical, and chemical conditions on the seafloor, which need to be monitored over time to assess the process and the time taken for restoration of environmental conditions. 相似文献
19.
Vance P. Vicente 《Marine Ecology》1989,10(2):179-191
Abstract. Long-term changes in the distributional patterns of commercial sponges ( Spongia spp. and Hippospongia spp.) within the West Indian Region indicates that: 1) commercial sponges had a widespread distribution throughout the whole West Indian Region and were ubiquitous in very shallow water until about the first half of the present century; 2) they were fished commercially not only in the traditional northern Caribbean sites (Florida, Gulf of Mexico, Bahamas) but also in the Greater ( e. g. , Hispaniola, Jamaica) and Lesser Antilles; and 3) they became extinct throughout most of the Lesser Antilles ( e. g. , Puerto Rico, Vieques, St. Thomas) sometime during the first half of this century. Mortalities of spongiids within the Antilles were found to differ from other marine mortalities in that: 1) species disappeared from a large region; 2) species vanished from different habitats and depths; and 3) natural populations never recovered. Species richness patterns suggest that commercial sponge genera ( Spongia and Hippospongia ) evolved under slightly cooler elimatic conditions than those found at present, and that these extinctions occurred as a direct or indirect effect of positive thermal anomalies in sea surface and atmospheric temperatures between 1900–1950. The concept that species diversity is stable on a regional scale is questioned. 相似文献
20.
The behaviour of an axially pre-loaded cylindrical member of an offshore structure, hit by a supply ship, has been investigated. The effects of axial pre-loading on the dynamic properties of members, the extent of damage and propagation of dynamic instability in the tubular members have also been investigated. Numerical models have been validated using available experimental data from the literature although these are mostly static with no pre-loading. Axial pre-loading was found to change the dynamic characteristics of some cylindrical members quite dramatically while it had no significant effect on others. The study examined whether there was any change in the behaviour between local and global modes when the loading was dynamic rather than static. The effect of damping on the dynamic instability of axially pre-loaded tubes under lateral dynamic loads was also studied. It was observed that for some geometries the quasi-static response can be used to define the boundary between bounded and unbounded dynamic responses. The main contribution of this study, compared with previous investigations reported in the literature, is that both pre-loading and dynamic effects have been included. 相似文献