共查询到19条相似文献,搜索用时 62 毫秒
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鲤浮肿病(carp edema virus disease,CEVD)是在锦鲤中发现的一种传染性病毒病,近年来在全球传播迅速,对鲤养殖业造成严重损害。本研究设计CEV环介导等温扩增(loop-mediated isothermal amplification,LAMP)引物,进行特异性实验;选取套式PCR、荧光PCR和LAMP三种检测方法,组织10家实验室进行室间比对测试:制备含CEV病毒核酸的测试样品,评价其均匀性和稳定性,通过测试结果分析方法检出限。结果显示,套式PCR的核酸样品检测限可达6.8×10-10 g。 相似文献
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本研究针对鲤浮肿病病毒(carp edema virus, CEV)基因组核蛋白编码基因P4a的序列,设计2对特异性引物,以克隆构建的重组质粒为标准模板,通过优化反应体系中引物浓度组合、Mg~(2+)浓度、dNTPs浓度、反应温度和扩增时间等参数,建立了CEV环介导等温扩增(loop-mediated isothermal amplification,LAMP)检测方法。结果表明,CEV-LAMP方法的最佳反应温度为62℃,引物浓度组合为引物F3/B3 0.2μmol/L,引物FIP/BIP 1.2μmol/L, Betaine0.7 mol/L, Mg~(2+) 8.0 mmol/L, dNTPs 1.2 mmol/L,反应时间60 min。反应产物经凝胶电泳呈现梯型条带,添加SYBR Green I荧光染料后,呈现明显的绿色阳性反应。CEV-LAMP法灵敏度高,最低检测限为10 copies/μL,较常规PCR法灵敏度高100倍; CEV-LAMP法特异性强,与锦鲤疱疹病毒(KHV)、鲤疱疹病毒Ⅱ型(CyHV-2)、鳜传染性脾肾坏死病毒(ISKNV)及鲤春病毒血症病毒(SVCV)无交叉反应。CEV-LAMP应用于患病鲤样本检测结果准确,简便快速,可为鲤浮肿病的现场诊断与防控提供技术支撑。 相似文献
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Snježana Zrnčić Dražen Oraić Ivana Giovanna Zupičić Željko Pavlinec Dragan Brnić Žaklin Acinger Rogić Ivica Sučec Dieter Steinhagen Mikolaj Adamek 《Journal of fish diseases》2020,43(6):673-685
Common carp (Cyprinus carpio) is a very important fish species for warm-water aquaculture in Croatia. All Croatian carp farms are subjected to a surveillance programme for the presence of koi herpesvirus (KHV), causing a deadly disease called koi herpesvirus disease (KHVD). However, there is no surveillance for other viral pathogens of importance like carp edema virus (CEV), a causative agent of koi sleepy disease (KSD). During regular testing within the KHVD surveillance programme, we tested samples for CEV simultaneously. The screening indicated possible outbreaks of KHVD and KSD. During 2016, KHVD broke out in an isolated area and soon thereafter a KHV eradication programme was successfully performed. However, during 2018 and 2019, two additional mortality events occurred in lakes in the southern part of Croatia during the spring. Samples from both events tested positive for CEV. An epidemiological investigation confirmed the introduction of infected carps from an infected farm to one of the lakes. To prevent the spreading of CEV into open waters, it is of utmost importance to introduce CEV testing before fish movement or to perform regular testing of all carp farms in the country to determine CEV prevalence for the purpose of implementation of control measures. 相似文献
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D Steinhagen 《Journal of fish diseases》2018,41(3):463-468
Hungary is an important carp producer with intensive trading relationships with farms in other carp‐producing areas in Europe. Carp in Europe were recently found infected with carp edema virus (CEV), a poxvirus which causes the koi sleepy disease (KSD) syndrome. Moribund carp were collected from 17 fish farms and angling ponds in different regions of Hungary. Histological analysis of gills from these carp revealed a proliferation of the interlamellar epithelium and an infiltration by eosinophilic cells. In 13 of 17 of these carp, CEV DNA was detected by qPCR and in seven fish more than 1 × 104 copies of virus‐specific DNA sequences per 250 ng of DNA, which could be considered as clinically relevant and a cause of disease. A phylogenetic analysis of the sequences revealed that all three genogroups of CEV were present in Hungarian common carp with genogroup I being most abundant. These results support the hypothesis of a prolonged presence of CEV in European carp populations and suggest that previous outbreaks of KSD were not recorded or misdiagnosed. Hence, a testing of carp and koi for infection with CEV should be included into disease surveillance programmes to prevent further spreading of this disease. 相似文献
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Carp edema virus in Polish aquaculture – evidence of significant sequence divergence and a new lineage in common carp Cyprinus carpio (L.) 下载免费PDF全文
M Matras E Borzym D Stone K Way M Stachnik J Maj‐Paluch M Palusińska M Reichert 《Journal of fish diseases》2017,40(3):319-325
Fish samples initially collected by local veterinarians on the common and koi carp farms in Poland between 2013 and 2015 as part of a KHV surveillance programme, when the water temperature was between 16 and 26 °C, and were also tested for CEV by qPCR. A partial 478 nucleotide fragment of the 4a gene was subsequently generated from 17 qPCR‐positive common carp Cyprinus carpio samples from 36 farm sites tested during the period. Sequence alignments and analysis revealed the presence of CEV in Poland both in common carp as well as in koi carp farms, and phylogenetic analysis assigned the Polish CEV sequences into three distinct genogroups. A lineage which includes the original sequences obtained from koi carp in Japan (genogroup II) included sequences from both koi carp and common carp, and the second lineage (genogroup I) contained sequences from common carp only. A third lineage (genogroup III) which was more closely related to the genogroup II also consisted of sequences from common carp only. The latter represents a lineage of CEV not previously described in the literature. 相似文献
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在23~25℃下,给平均体质量104.2g和106.8g的回交鲤(鲤鲫杂交♀×鲤♂)和回交荷包红鲤(鲤鲫杂交♀×荷包红鲤♂)胸腔注射植物凝集素(PHA)和秋水仙素,然后观察对比了染色体核型,探讨回交后代的染色体遗传。结果表明,两种鲤鲫杂交回交子代染色体组均由150条染色体组成,按着丝粒位置染色体组型可分为四组,每个染色体小组均由三条同源染色体组成。其中,回交鲤染色体数目为3n=150,染色体臂数NF=234,其核型公式为:3n=60m+24sm+36st+30t;回交荷包红鲤的染色体数目为3n=150,染色体臂数NF=252,其核型公式为:3n=66m+36sm+18st+30t。 相似文献
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Soumesh K. Padhi Isaiah Tolo Margaret McEachran Alexander Primus Sunil K. Mor Nicholas B. D. Phelps 《Journal of fish diseases》2019,42(11):1609-1621
Koi herpesvirus (KHV; cyprinid herpesvirus‐3) and carp oedema virus (CEV) are important viruses of common and koi carp (Cyprinus carpio); however, the distribution of these viruses in wild common carp in North America is largely unknown. During the summers of 2017 and 2018, 27 mass mortalities of common carp were reported from four states in the USA (Minnesota, Iowa, Pennsylvania and Wisconsin), the majority of which were distributed across eight major watersheds in southern Minnesota. Samples from 22 of these mortality events and from five clinically healthy nearby carp populations were screened for KHV, CEV and SVCV using real‐time polymerase chain reaction (qPCR). KHV was confirmed in 13 mortality events, CEV in two mortality events and coinfections of KHV/CEV in four mortality events. Nucleotide sequence analysis revealed that the KHV and CEV detected here are closely related to European lineages of these viruses. While molecular detection alone cannot conclusively link either virus with disease, the cases described here expand the known range of two important viruses. This is also the first reported detection of KHV and CEV coinfections in wild carp populations. 相似文献
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Experimental infection of European crustaceans with white spot syndrome virus (WSSV) 总被引:3,自引:1,他引:3
V Corbel Z Zuprizal C Shi Huang Sumartono J-M Arcier & J-R Bonami 《Journal of fish diseases》2001,24(7):377-382
Eight European marine and freshwater crustaceans were experimentally infected with diluted shrimp haemolymph infected with white spot syndrome virus (WSSV). Clinical signs of infection and mortalities of the animals were routinely recorded. Diagnosis was by direct transmission electron microscopy (TEM), DNA hybridization (dot-blot and in situ hybridization) using WSSV probes and by PCR using WSSV specific primers. High mortality rates were noted between 7 to 21 days post-infection for Liocarcinus depurator , Liocarcinus puber , Cancer pagurus , Astacus leptodactylus , Orconectes limosus , Palaemon adspersus and Scyllarus arctus . Mortality reached 100%, 1 week post-infection in P. adspersus . When infection was successful, direct TEM observation of haemolymph revealed characteristic viral particles of WSSV, some observed as complete virions (enveloped), others as nucleocapsids associated with envelope debris. WSSV probes showed strong positive reactions in dot-blots and by in situ hybridization in sections and specific virus DNA fragments were amplified successfully with WSSV primers. White spot syndrome virus was pathogenic for the majority of the crustaceans tested. This underlines the epizootic potential of this virus in European crustaceans. 相似文献
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Gill‐associated virus (GAV) was found to be associated with decreased prawn, Penaeus monodon, production when prawns from three farms (n = 45 ponds, 1800 prawns) were monitored for GAV over the production season using a graded RT‐nPCR. The grading system used was a visualization of either the outer or inner nested PCR products. Prevalence and loading of GAV were associated with disease severity. Ponds with a higher initial prevalence and a larger increase in GAV load over the production period suffered disease outbreaks. Ponds with low initial prevalence of GAV but a larger increase in prevalence and large increase in load over the production period suffered chronic disease with no disease outbreak identified, yet low production. However, the ponds with moderate to low initial prevalence of GAV with a low increase in prevalence and load of GAV over the production period incurred no disease outbreak and comparatively high production. Ponds with GAV prevalence greater than 75% at 1 month post‐stocking should be considered for termination as they have a high probability (95%) of having a disease outbreak. Emergency harvest when a disease outbreak occurs will significantly limit mortality losses. 相似文献
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在饲料中添加柑橘或甜橙外果皮对鲤鱼诱食作用的比较研究 总被引:11,自引:0,他引:11
在基础饵料中不同量(0.05%、0.1%、0.25%、0.5%、1%、3%)的柑橘或甜橙外果皮,用记录仪记录鲤鱼啄咬饵料球的频率和力。表明,柑橘和甜橙外果皮是2种新的鲤鱼诱食物物质,而且二者对鲤诱食作用的效果基本相同,其适宜添加量分别为0.25%或0.1%。 相似文献