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A loop-mediated isothermal amplification (LAMP) procedure is described for rapid diagnosis of white tail disease, a viral disease caused by Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV), in the giant freshwater prawn, Macrobrachium rosenbergii. This method was more sensitive than conventional RT-PCR for detecting the two viruses. A set of four primers, two outer and two inner, were designed for MrNV detection. An additional pair of loop primers was also used in an accelerated LAMP reaction for detection of XSV. Time and temperature conditions were optimized for detection of the two viruses. The LAMP reaction is highly suited for disease diagnosis in developing countries as amplification of DNA can be detected without the use of agarose gel electrophoresis, by the production of whitish precipitate of magnesium pyrophosphate as a by-product. 相似文献
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Qian D Shi Z Zhang S Cao Z Liu W Li L Xie Y Cambournac I Bonami JR 《Journal of fish diseases》2003,26(9):521-527
A disease of Macrobrachium rosenbergii, the giant freshwater prawn, farmed in China was recently recorded in Zhejiang, Jiangsu, Shanghai, Guangxi and Guangdong provinces. The clinical sign of the disease, which develops in post-larvae (PL), is a whitish appearance of the muscles, particularly noticeable in the abdomen. Mortalities may reach 100% in some hatcheries. Investigations by transmission electron microscopy after negative staining of diseased PL homogenates showed the presence of two types of viral particles: one, unenveloped, icosahedral in shape, 26-27 nm in diameter, the second, much smaller, about 14-16 nm in diameter, designated extra small virus particle (XSV). The large virus has a genome with two pieces of ssRNA (RNA-1 and RNA-2), of 3 and 1.2 kb, respectively. Hybridization tests confirmed that this large virus is closely related to M. rosenbergii nodavirus (MrNV) which was isolated from diseased prawns in a hatchery in the French West Indies. Its very small size and hypothesized biochemical and biological characteristics suggest XSV is a new type of crustacean virus. As XSV has always been found associated with the larger virus (nodavirus) and is located in muscle and connective cells of diseased animals, it could be an autonomous virus, a helper-type virus or a satellite-like virus. 相似文献
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为建立罗氏沼虾育种项目,2006年利用罗氏沼虾3个群体,通过巢式交配设计,实现了大规模构建罗氏沼虾父系半同胞家系。实验中,每箱放置亲虾1雄5雌,按交配设计共布置100个交尾网箱;通过对培育条件及幼体数量的标准化,使每个家系在幼体培育的各个阶段的条件尽量保持一致,减少由于条件不一致造成的家系间的环境偏差。结果显示,500尾雌虾抱卵虾为271尾,亲虾抱卵率平均为54.2%,且每个网箱出现两尾以上抱卵虾的网箱数为92个,占全部网箱数的92%;通过标准化培育,最终建立了123个家系,其中含有父系半同胞家系37个。本试验结果为建立罗氏沼虾全同胞和半同胞家系提供了依据。 相似文献
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This study was performed to assess the nutritional value of Streptocephalus dichotomus nauplii and compared with standard larval diet and parthenogenetic strain of Artemia nauplii. The effectiveness of live feed was determined by feeding freshwater prawn Macrobrachium rosenbergii postlarvae. Results on the growth rate, weight gain, survival, fatty acid and amino acid composition show a significant variation between the dietary treatments. Mean larval growth was significantly different ( P < 0.001) between control diet (13.5 ± 0.5 mm) and live feeds ( Streptocephalus nauplii; 18.4 ± 0.5 mm and Artemia nauplii 18.7 ± 0.2 mm). The weight of larvae-fed Streptocephalus (41.1 ± 1.44 mg) and Artemia nauplii (41.7 ± 0.2 mg) was not significantly different; however, treatment with live feeds was significantly higher than the control (16.3 ± 0.5 mg) ( P < 0.001). Proximate composition on the fairy shrimp reveals that they are rich in protein, lipid, essential amino acids (EAA) and essential fatty acids (EFA). The polyenoic unsaturated fatty acid (18 : 2 n-6 and 18 : 3 n-3) and highly unsaturated fatty acid (20 : 4 n-3 and 20 : 5 n-3) show a dramatic increase in larval tissue relative to its proportional composition in the live diets. Amino acid composition in the live feeds, Streptocephalus and Artemia nauplii, perfectly reflects on the amino acid proportions in the larvae-fed diets which confirm its utilization. 相似文献
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A sandwich enzyme linked immunosorbent assay (S-ELISA) for detection of MrNV in the giant freshwater prawn,Macrobrachium rosenbergii (de Man) 总被引：2，自引：0，他引：2
A sandwich enzyme-linked immunosorbent assay (S-ELISA) was developed to improve diagnosis of white tail disease of the giant freshwater prawn, Macrobrachium rosenbergii, caused by the nodavirus, MrNV. Polyclonal antibodies were produced by immunization of Balb/C mice using a purified suspension of the virus and IgG anti-MrNV were purified from ascitic fluid. A sandwich method was successfully developed, coating first with unlabelled antibody and detecting trapped antigens with a second biotinylated antibody. Reaction was demonstrated using an avidin-peroxidase conjugate. Tissue extracts from M. rosenbergii infected with MrNV or purified viral extracts (control) were successfully identified in an individual ELISA, thus confirming the validity of the method. This S-ELISA should be the technique of choice for epidemiological studies of this disease and is a rapid and inexpensive assay with high specificity and sensitivity. 相似文献
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Sri Widada J Durand S Cambournac I Qian D Shi Z Dejonghe E Richard V Bonami JR 《Journal of fish diseases》2003,26(10):583-590
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罗氏沼虾不同养殖条件下的生长和存活率相关分析 总被引：1，自引：0，他引：1
为探讨不同养殖条件下罗氏沼虾重要经济性状的相关性,以106个罗氏沼虾家系为材料,将每个家系随机分成两组,分别放入两个大小不同的池塘中,以不同的养殖密度养殖,两个月后测定每个家系的头胸甲长、腹长、体长、体重及存活率,并以家系为单位进行同一性状在两种不同养殖环境条件下的相关分析。结果表明,罗氏沼虾头胸甲长、腹长和体长在两种不同养殖环境条件下的对应性状间均呈不显著正相关(P>0.05),相关系数依次为r=0.109、r=0.15和r=0.143;体重呈显著正相关(r=0.223,P<0.05);存活率呈极显著正相关(r=0.862,P<0.01)。头胸甲长、腹长和体长的变异系数范围在10.85～18.09之间;两个土池中体重的变异系数分别为36.80和43.47,说明罗氏沼虾体重性状具有较大的遗传选择潜力。研究结果表明,罗氏沼虾在不同养殖条件下,以体重、存活率为选育指标,尽管选择趋势不变,但选种效果有差别。 相似文献
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Immunomodulatory effect of Cynodon dactylon against white tail disease of giant freshwater prawn,Macrobrachium rosenbergii (de Man, 1879) 
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下载免费PDF全文 Mohamed A Farook Sugumar Vimal Nithiyanandam Madan Gani Taju Seepoo Abdul Majeed Kalaiselvi S N Nambi Gnanavel Balasubramanian Azeez S Sahul Hameed 《Aquaculture Research》2016,47(11):3421-3431
The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important and extensively cultured crustacean worldwide. The viral pathogens, Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) are responsible for causing severe mortalities in the hatchery and nursery phases. This study investigates the protection of postlarvae of freshwater against white tail disease (WTD) using plant extract derived from Cyanodon dactylon and the modulation of the prawn non‐specific immunity. To determine the immunomodulatory effect of C. dactylon extract, the prawn was injected with plant extract and various immunological parameters were estimated. The immunological parameters such as proPO, SOD, THC and clotting time were found to be significantly higher in the plant extract‐injected prawn when compared with control groups. The results of real time PCR analysis revealed up regulation on the expression proPO, SOD and lysozyme genes in MrNV and XSV challenged prawn postlarvae treated with C. dactylon extract. Infectivity experiment showed high relative per cent survival in MrNV and XSV‐challenged prawn postlarvae treated with C. dactylon extract. These results strongly indicate that the administration of C. dactylon plant extract enhances immunity of the prawn. Based on the results, this study recommends that the immersion of postlarvae in C. dactylon plant extract is a potential prophylactic agent against WTD. 相似文献
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A feeding experiment was conducted to investigate the effect of Bacillus subtilis bacterium, on larval growth and development rate of Macrobrachium rosenbergii (de Man) during February 28 to April 8, 2005 in University Putra Malaysia hatchery. Newly hatched larvae of M. rosenbergii were reared with two dietary treatments consisting of newly hatched Artemia salina nauplii with B. subtilis (108 cells ml−1), and newly hatched A. salina nauplii without B. subtilis carried out in triplicate in 60‐L aquarium (50 L−1). After trial, the larvae that fed B. subtilis‐treated Artemia naupli were found to have higher survival and a faster rate of metamorphosis than larvae that were fed with nontreated Artemia naupli. There were significant differences between B. subtilis‐treated Artemia naupli and nontreated Artemia diet in larval growth and development rate of metamorphosis (P < 0.05). Larval survival after 40 days was significantly greater (P < 0.05) in the B. subtilis‐treated groups (55.3 ± 1.02) compared with the nontreated groups (36.2 ± 5.02%). 相似文献
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Larvae of two caridean shrimp species, Macrobrachium rosenbergii (De Man) and Palaemon elegans Rathke, were fed live and artificial diets. P. elegans larvae fed exclusively live Artemia salina (15 nauplii mL?1) developed into first postlarval stage (PL1) within 12 days at a temperature of 25°C and salinity 32.5 g L?1. Their survival and mean total length at this stage were 88.5% and 6.7 mm respectively. M. rosenbergii larvae fed on 15 Artemia mL?1 started to metamorphose into PLl within 24 days at 29–30°C and 12 g L?1. Attempts to completely replace live Artemia for rearing P. elegans during early stages failed, and only a partial replacement was achieved for the larvae of both species. P. elegans larvae survived (49%) solely on a microgranulated diet (Frippak PL diet) from stage zoea (Z) 4–5 to PL1. Similarly, a microencapsulated diet (Frippak CD3) also sustained M. rosenbergii larvae from Z5–6 to PL1 with a 28% survival. Development of the larvae of both species was retarded by 2–3 days and their survivals were lower than those fed on the live diet. The inability of the early larvae of these caridean species to survive on artificial diets is attributed to their undeveloped guts and limited enzymatic capabilities. Trypsin activity in the larvae was determined for all larval stages. It was found that the highest trypsin activity, at stage Z4–5 in P. elegans and at stage Z5–6 in M. rosenbergii, coincides with a rapid increase in the volume of the hepatopancreas and the formation of the filter apparatus. These morphological changes in the gut structure appear to enable the larvae to utilize artificial diets after stage Z5–6. Low larval trypsin activities may be compensated by the easily digestible content of their live prey during early larval stages (Z1–Z4/5) and by longer gastroevacuation time (GET) and almost fully developed guts during later stages. 相似文献
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White tail disease of the farmed freshwater prawn, Macrobrachium rosenbergii, is the cause of mortalities in the French West Indies, China and India. Two different sized particles, both developing in the cytoplasm of target cells, are found associated with diseased animals. These two viruses were separated, purified and subsequently characterized. The larger one, called MrNV, is icosahedral in shape and 27 nm in diameter. Its genome is composed of two fragments of linear single-stranded RNA (ss-RNA), of 2.9 and 1.3 kb, respectively and its capsids exhibited a single polypeptide of 43 kDa. These characteristics and the partial sequence of a cloned fragment of RNA-1 suggest this agent is a member of the family Nodaviridae, but with differences from both the genera Alphanodavirus and Betanodavirus. The smaller virus, named XSV, is icosahedral in shape, 15 nm in diameter, possesses a linear ss-RNA genome of about 0.9 kb, and its capsid exhibits two polypeptides of 16 and 17 kDa, respectively. The relationships between these two viruses remain unknown. 相似文献
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Macrobrachium rosenbergii nodavirus (MrNV) that causes white tail disease (WTD) is an emerging disease that contributes to serious production losses in Macrobrachium hatcheries worldwide. Mosquito cell lines (C6/36) have been reported to support the growth of MrNV and used to observe the cytopathic effects (CPE) in infected cells. This study determined the susceptibility of C6/36 mosquito cells to the Australian isolate of MrNV in order to use fewer animals in further investigations. Different staining methods were used to observe MrNV viral activity in C6/36 cells. Typical cytopathic effects such as vacuolation and viral inclusion bodies were observed in infected C6/36 cells with H&E and Giemsa staining. With acridine orange, it was easier to detect presumptive MrNV messenger ribonucleic acid in the infected cells. Using neutral red staining to measure mitochondrial activity showed light absorption of infected cells maximized at day 4 (O.D. = 0.6) but was significantly lower (chi‐square = 41.265, df = 1, P < 0.05) than control groups (O.D. = 2) which maximized at day 12. Using trypan blue staining to count the number of cells with disrupted cell membranes, the maximum number of presumptively dead cells at day 8 (4 × 105 cells) in infected treatments was higher than the control treatment at day 10 (1.8 × 105 cells). However, TaqMan real‐time PCR did not confirm the replication of MrNV in the cells over 14 days. The mean viral copies and mean cycle times of positive samples were stable at 2.07 × 104 and 24.12, respectively. Limited evidence of viral replication was observed during four serial passages. This study determined the mortality of the C6/36 cell line to the Australian isolate of MrNV but suggests limited patent replication was occurring. Trying different cell lines or adapting the virus to the C6/36 cells may be necessary to successfully replicate Australian MrNV in cell lines. 相似文献
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为探索罗氏沼虾(Macrobrachium rosenbergii)铁虾综合征(IPS)的分子机制,采用高通量测序平台(Illumina Hiseq-2500)分别对患IPS罗氏沼虾(IPS虾)和正常罗氏沼虾开展转录组测序,进行生物信息学分析。结果显示,高通量测序共获得56.42 G高质量数据,拼接后得到221 901条单基因序列(unigene),长度范围为201~30 985 bp,平均长度为1572 bp,N50长度为2867 bp,N90长度为646 bp。将单基因序列分别在Nr、Nt、Swissprot、KEGG、KOG、GO、PFAM数据库进行序列比对及功能注释,103 570条得到注释,其中,GO数据库注释到的单基因序列最多。差异表达分析显示,2003个基因在IPS虾眼柄中差异表达,包括1209个上调基因和794个下调基因,516个基因被注释到242条KEGG通路中,翻译、信号转导和免疫系统富集的差异基因数目最多。催乳素、雌激素、胰岛素、促性腺激素释放激素、胰高血糖素、催产素、谷氨酸能突触、血清素能突触等与生殖调控相关激素的代谢过程在IPS虾与正常虾眼柄之间存在差异。此外,一些已被证明在免疫反应中起重要作用的基因在IPS虾眼柄中显著上调,如血管内皮生长因子受体1、丝氨酸蛋白酶抑制剂6、C型凝集素、芳基硫酸酯酶B、酚氧化酶原激活酶2a、组织蛋白酶B、组织蛋白酶L、甲壳类抗菌肽4等。同时,注释到溶酶体、吞噬体、抗原处理与呈递、细胞凋亡、内吞作用等多条与免疫相关的途径,支持近期研究得出的罗氏沼虾IPS与病原感染相关的结论。本研究为解析罗氏沼虾IPS的成因和分子机制提供了数据支撑。 相似文献
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对罗氏沼虾（Ｍａｃｒｏｂｒａｃｈｉｕｍｒｏｓｅｎｂｅｒｇｉ）冷冻调理品的加工方法和保鲜技术进行了研究。试验结果表明,加工后的罗氏沼虾,其保藏性能增强,加添加剂后罗氏沼虾的鲜度下降明显减慢。 相似文献