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根据派琴虫的基因保守序列设计了特异性引物,对派琴虫的DNA进行PCR扩增并将产物克隆到pMD18-T载体后测序.结果表明,扩增大小为596bp,与预期扩增序列同源性为99.8%.该PCR方法检测灵敏度高,最低可检测1pg的派琴虫DNA;特异性强,对荧光假单胞菌、嗜水气单胞菌、大肠杆菌、葡萄球菌、副溶血孤菌、沙门氏菌、诺瓦克样病毒等贝类病原体的扩增均为阴性.用该PCR技术对广西沿海的104份牡蛎、49份贻贝和20份文蛤病料进行检测,阳性率分别为14.6%、10.6%和15.0%.结果显示,派琴虫广泛存在于中国南方沿海的养殖贝类中,建立的PCR方法可用于贝类派琴虫的临床快速检测. 相似文献
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选择派琴虫保守的核糖体DNAITS-2区域设计引物和TaqMan探针,通过对反应体系和反应条件进行优化,建立了实时荧光定量PCR检测派琴虫的方法。所构建方法检测质粒模板DNA的动态范围为2.6×10^1～2.6×10^7拷贝,敏感度可检测到26拷贝质粒DNA,而且与包拉米原虫、隐孢子虫等其他寄生性原虫无交叉反应,也不受贝类组织DNA的干扰。利用本研究所建立的方法对来自我国山东、福建等不同沿海海域的30份贝类样品进行检测,检出阳性样品3份。研究表明,本研究所构建的派琴虫实时荧光定量PCR检测方法具有快速、灵敏和特异等优点,可满足国内养殖场及进出口水生动物携带派琴虫的检测需要。 相似文献
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于2011-2012年期间,对白洋淀水域4种经济鱼类(鲫鱼、草鱼、鲢鱼、鳙鱼)指环虫(Dactylogyrus)的寄生情况进行了调查,并对指环虫的重要分类特征——中央大钩和交接器进行了显微拍照。本次调查共发现指环虫12种(其中2种为未定种),在鲫鱼上发现6种,其中美丽指环虫(D.formosus)和弧形指环虫(D.arcuatus)为优势种。鲢鱼上发现2种,鲢指环虫(D.hypophalmichthys)为优势种。鳙鱼上发现2种,鳙指环虫(D.aristichthys)为优势种。草鱼上发现2种,页形指环虫(D.lamellatus)为优势指环虫。通过调查,初步掌握了白洋淀4种经济鱼类指环虫的寄生情况,为白洋淀指环虫病的研究提供了基础资料。 相似文献
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寄生虫对大黄鱼养殖造成的危害极大 ,因其种类多、不分季节、寄生的部位又很广泛 ,使人们防不胜防。现主要介绍几种 ,供养殖业者参考、借鉴。一、指环虫病1 .病原体 指环虫。2 .流行情况 主要危害大黄鱼的鱼种和成鱼 ,四季均可发生 ,寄生于鱼鳃内。3 .主要症状 轻度感染时症状不明显 ,当大量寄生时 ,由于指环虫以其钩及边缘小钩钩住寄生的鳃组织 ,并在鳃上不断蠕动 ,因而破坏鳃丝的表皮细胞 ,刺激鳃丝细胞分泌过多的粘液 ,妨碍鱼的呼吸并使鱼体贫血。在低倍镜下 ,可以观察到鲜活病鱼鳃片上正在伸缩活动的虫体。若一个视野虫体达 5… 相似文献
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根据GenBank中单孢子虫和折光马尔太虫基因序列,用Primer Express 2.0软件设计了两对引物和两条TaqMan探针.对反应条件和试剂浓度进行优化,建立了能够同时检测单孢子虫和折光马尔太虫的二重荧光定量PCR方法.该方法对单孢子虫和折光马尔太虫的检测敏感性达到40个模板拷贝数;此外抗干扰能力强,对单孢子虫和折光马尔太虫不同模板浓度进行组合,仍可有效地同时检测到这两个原虫.该方法对派琴虫、荧光假单胞菌、副溶血弧菌、溶藻弧菌、河弧菌和拟态弧菌等病原体的检测,结果全为阴性.研究建立的单孢子虫和折光马尔太虫荧光定量PCR具有特异、敏感、快速、定量、重复性好等优点,可用于贝类单孢子虫和折光马尔太虫感染的检测. 相似文献
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近年来,车轮虫病及其引起的继发性疾病在全国各地均有大面积发生。危害的品种很多,包括各种养殖经济鱼类以及贝类等。流行的季节较长,肉眼不易诊断,治疗难度大,容易复发。本文简要阐述车轮虫的寄生特点及该病的诊治现状,并就当前如何防治车轮虫病提出笔者个人意见。一、车轮虫的寄生特点1.车轮虫在宿主上的分布车轮虫属自建立以来已报 相似文献
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在阐述海水养殖贝类病害出现原因的基础上,提出海水养殖贝类病害预警预报技术,并以某海域的养殖贝类病害预警工作情况为例,分析基于海水养殖贝类病害预警预报技术的养殖贝类病害预警预报模型构建及应用,旨在能够为海水养殖贝类病害防范和解决提供重要参考支持. 相似文献
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奥尔森帕金虫是重要的贝类病原性寄生虫之一,为建立快速、灵敏、准确和使用简便的检测方法,实验根据奥尔森帕金虫5.8S rDNA中的内转录间隔区(internal transcribedspacer,ITS)序列,建立了环介导等温扩增(loop-mediated isothermal amplification,LAMP)检测方法,并对反应温度、反应体系中Mg2+浓度和反应时间进行了优化。该方法的检测灵敏度约为30拷贝质粒DNA,并且特异性较强,与海水帕金虫、包纳米虫、波豆虫及急性病毒性坏死病毒(acute viral necrosis virus,AVNV)等病原均无交叉反应。使用LAMP法对两批菲律宾蛤仔样品进行检测,结果表明,LAMP检测与传统PCR检测相比,灵敏度更高,检测结果更准确。实验所建立的奥尔森帕金虫LAMP检测方法简单、快速、灵敏且特异性强,可以在沿海贝类养殖厂及条件简陋的实验室使用。 相似文献
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对中国沿海23个地点的太平洋牡蛎（Crassostrea gigas）、近江牡蛎（Crassostrea ariakensis）、褶牡蛎（Ostrea plicatula）进行样品采集。利用液体巯基醋酸盐培养基培养法、组织切片法、聚合酶链反应法,对样品进行帕金虫含量检测。结果表明在三种牡蛎体内春、夏、冬季都有帕金虫寄生,夏季帕金虫感染率最高可达100%,其他季节感染率较低。利用分子生物学方法鉴定帕金虫种类,结果表明,在中国沿海的三种牡蛎体内主要寄生两种帕金虫：Perkinsus.sp.n（EU068107）与Perkinsus olseni,这两种帕金虫在中国沿海分布具有一定地域性,Perkinsus.sp.n（EU068107）主要分布在中国南海的海南省、广西壮族自治区、广东省沿岸,而Perkinsus olseni分布在中国东海的浙江省沿岸。 相似文献
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A novel sialic acid-specific lectin (MCsialec) was detected from an expressed sequenced tag (EST) sequence from Manila clam haemocytes infected with Perkinsus olseni. The cDNA of the lectin was cloned using gene-specific primers based on a previously determined EST and characterized. The full-length cDNA of MCsialec is 603 bp in length and encodes a polypeptide of 200 amino acids with a calculated molecular mass of 21.928 kDa. Sequence alignment and protein motif analyses showed that MCsialec shares identity with sialic acid-specific invertebrate lectins from Cepaea hortensis, Helix pomatia and Haliotis discus discus. The lectin was expressed in Escherichia coli M15 cells and purified using a Ni-NTA His-binding resin matrix for antibody production. The presence of the lectin in various tissues of Perkinsus-infected and uninfected Manila clams was analysed by both PCR and immunohistochemical localization assays. MCsialec was detected in each tissue of the clams; however, upon infection, the level of expression of the lectin increased in each tissue. Vibrio tapetis infection also induced high-level expression of MCsialec in the haemocytes. These data suggest that MCsialec plays a crucial role in the immune system of the Manila clam during pathogenic infection. 相似文献
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Ana C. Luz Cunha Vitor de A. Pontinha Maria Alcina M. de Castro Simone Sühnel Sthefanie C. Medeiros ngela M. Moura da Luz Ricardo Harakava Leonardo Tachibana Danielle F. Mello Naissa M. Danielli Alcir L. Dafre Aimê R. M. Magalhes Jos Luiz P. Mourio 《Journal of fish diseases》2019,42(3):455-463
Perkinsus spp. have been detected in various bivalve species from north‐east Brazil. Santa Catarina is a South Brasil state with the highest national oyster production. Considering the pathogenicity of some Perkinsus spp., a study was carried out to survey perkinsosis in two oyster species cultured in this State, the mangrove oyster Crassostrea gasar and the Pacific oyster Crassostrea gigas. Sampling involved eight sites along the state coast, and oyster sampling was collected during the period between January 2013 and December 2014. For the detection of Perkinsus, Ray's fluid thioglycollate medium (RFTM) and histology were used, and for the identification of the species, PCR and DNA sequencing were used. Perkinsus spp. was found by RFTM in C. gigas and C. gasar from São Francisco do Sul. This pathology was also detected in C. gasar from Balneário Barra do Sul both, by RFTM and histology. Perkinsus marinus was identified in C. gigas and C. gasar from São Francisco do Sul and Perkinsus beihaiensis in C. gasar from Balneário Barra do Sul. This is the first report of P. marinus in C. gigas from South America. Results of this preliminary study suggest that both oyster species tolerate the species of Perkinsus identified, without suffering heavy lesions. 相似文献
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Ricardo B. Leite Ricardo Afonso M. Leonor Cancela 《Aquaculture (Amsterdam, Netherlands)》2004,240(1-4):39-53
This study characterises Perkinsus atlanticus disease prevalence and intensity within the economically important beds of the clam Ruditapes decussatus along the Iberian Atlantic coast. Samples were collected from five different sites along the Portuguese coast (Ria Formosa, Ria do Alvor, V.N. Milfontes, Aveiro, Lagoa de Obidos) and in Galicia (Ria de Arosa) between winter 2000/2001 and 2002/2003. The infection level was evaluated by the Ray Fluid Thioglycollate medium (RFTM) method using the body burden assay. In addition, the measure of the condition index (CI) (percentage between the edible part and the total weight of the clams) was conducted to investigate a possible correlation between this parameter and the intensity of the infection. However, no clear relationship between infection intensity and condition index (CI) could be determined although a significant decline in CI was found for heavily infected clams. Results of a 2-year survey demonstrate the presence of Perkinsus sp. in all sampling sites albeit with different intensities. Sites where human interference was minimal showed the lowest levels of infection. There were no significant differences in Perkinsus sp. infection intensity between samples collected in winter and summer, in contrast with several studies describing higher intensities at the end of the summer. Nevertheless, major differences were observed from year to year and site to site, indicating that factors other than those responsible for seasonal climatic variations might affect the prevalence and the intensity of Perkinsus sp. infection. 相似文献
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Julie D. Gauthier Thomas M. Soniat James S. Rogers 《Journal of the World Aquaculture Society》1990,21(2):105-115
Oysters were collected along salinity gradients within four major oyster producing watersheds in Louisiana (Lake Calcasieu, Terrebonne Bay, Barataria Bay and Lake Borgne) and examined for parasites in relation to oyster condition and reproductive state. The percentage of Infection by Perkinsus marinus, Nematopsis prytherchi, N. ostrearum, Bucephalus cuculus , prokaryotic inclusion bodies, and ciliates was recorded for each site. Average pigment cell number, oyster condition, gonadal index, and the percentage of oysters with atrophic digestive diverticula were also determined.
In general, oysters experienced higher levels of parasitism in higher salinity waters. Perkinsus marinus Occurrence was correlated with salinity, an atrophied condition of the oysters, and Nematopsis spp. occurrence. Oysters in poor condition had higher concentrations of pigment cells, greater atrophy of the digestive diverticula, and increased parasitism by N. ostrearum . 相似文献
In general, oysters experienced higher levels of parasitism in higher salinity waters. Perkinsus marinus Occurrence was correlated with salinity, an atrophied condition of the oysters, and Nematopsis spp. occurrence. Oysters in poor condition had higher concentrations of pigment cells, greater atrophy of the digestive diverticula, and increased parasitism by N. ostrearum . 相似文献
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广东部分岛屿紫菜物种研究 总被引：1，自引：0，他引：1
为了解广东沿海紫菜物种自然资源现状及物种间的亲缘关系,于2012年冬季对广东3个岛屿进行调查,并对采集样品进行形态、细胞学以及基于rbcL和nrSSU序列的系统学研究。结果显示,12月平屿岛优势种有坛紫菜（Porphyrahaitanensis）、长紫菜（Pdentata）和皱紫菜（P．crispata）;外伶仃岛、下川岛为皱紫菜和多枝紫菜（Pramosissima）;3个岛屿的紫菜属于暖温性的刺缘组紫菜,染色体均为n=5,其中多枝紫菜染色体为首次观察报道。系统学结果显示,平屿岛皱紫菜与坛紫菜亲缘关系最近,与多枝紫菜之间的关系最远;外伶仃岛、下川岛皱紫菜与平屿岛皱紫菜的相似度远低于与多枝紫菜种间的系数。研究扩增了皱紫菜的nrSSU序列和多枝紫菜的rbcL、nrSSU序列,为紫菜的系统学研究提供了基础资料。 相似文献
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南麂列岛海洋自然保护区的虾类种类组成和数量分布 总被引：1，自引：0，他引：1
根据2013年11月、2014年2月、5月、9月在南麂列岛海洋自然保护区开展渔业资源调查所获得的虾类调查资料,将生物量作为虾类资源分布的数量指标,对南麂列岛海洋自然保护区的虾类种类组成、数量分布、优势种变化,及其与环境因子的关系等进行了研究。结果如下:(1)调查海域共鉴定出虾类25种,隶属于9科17属,虾类生物量各季节由高到低依次为夏季(32358.0 g)、秋季(13033.0 g)、冬季(3938.6 g)、春季(3635.6 g);(2)虾类数量分布季节变化明显,且岛礁区偏外的开阔海域虾类生物量比较高;(3)不同季节优势种更替较显著,春季优势种为日本鼓虾、鲜明鼓虾和细巧仿对虾,夏季优势种为哈氏仿对虾和中华管鞭虾,秋季优势种为中华管鞭虾、细巧仿对虾和哈氏仿对虾,冬季的优势种为细巧仿对虾、脊尾白虾、鲜明鼓虾和日本鼓虾;(4)水深对虾类生物量分布影响明显,各季节虾类生物量与环境因子相关性关系变化较大。结论认为,南麂列岛调查海域的虾类种类以季节性的广温广盐性种类为主;主要优势种生物量夏秋两季明显高于冬春两季,可能是因为大量虾类冬春季离开该海域去其他海域越冬繁殖,而到了夏秋季则回到该海域产卵、育肥和栖息;在夏、秋、冬季沿岸区虾类生物量高于岩礁区,可能因为虾类一般生活于有利于索饵、成长的泥沙底质沿岸区;优势种更替显著与主要优势种生长周期与虾类个体大小特征有关;南麂列岛海域虾类生物量与环境因子关系复杂与该海域多变的温度盐度、复杂区系特点有关;虾类生物量分布随水深变化分布明显,这主要是由于调查海域主要优势种虾类活动范围与水深有关。 相似文献