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J R Bonami - One of the best experts on this subject based on the ideXlab platform.
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Screening the post-larvae of Macrobrachium rosenbergii for early detection of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) by RT–PCR and immunological techniques
Aquaculture, 2011Co-Authors: A.s. Sahul Hameed, M. Ravi, G. Taju, M.a. Farook, R.i. Hernandez-herrera, J R BonamiAbstract:Abstract White tail disease (WTD) was found to be a serious problem in hatcheries and nursery ponds of freshwater prawn ( Macrobrachium rosenbergii ) . The causative organisms have been identified as Macrobrachium rosenbergii nodavirus ( Mr NV) and extra small virus (XSV). RT–PCR and immunological techniques such as Western blot and ELISA were used for early detection of Mr NV and XSV in post-larval samples obtained from time-course experiments at different time intervals. Two viruses were purified from diseased post-larvae of M. rosenbergii by a combination of low and high speed centrifugation using sucrose and CsCl gradients to raise the antisera separately. One structural protein with molecular weight of 43 kDa (CP-43) was identified from the purified preparation of Mr NV, and two overlapping polypeptides of about 17 kDa (CP-17) and 16 kDa (CP-16) were found in XSV particles by SDS-PAGE. The antisera raised against CP-43 of Mr NV, CP-16 and CP17 of XSV in mice were used to detect Mr NV and XSV by Western blot and ELISA. Published primers specific to Mr NV and XSV were used for the early detection of these viruses by RT–PCR and nested RT–PCR. The post-larval samples collected at 3 h post infection (h p.i.) showed positive for both viruses by nested RT–PCR and negative by RT–PCR, Western blot and ELISA techniques. The samples collected at 24 h p.i. and thereafter were found to be positive for Mr NV and XSV by RT–PCR, ELISA and Western blot analyses.
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rapid detection of Macrobrachium rosenbergii nodavirus mrnv and extra small virus xsv the pathogenic agents of white tail disease of Macrobrachium rosenbergii de man by loop mediated isothermal amplification
Journal of Fish Diseases, 2006Co-Authors: D Pillai, J R Bonami, Sri J WidadaAbstract:: 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.
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studies on the occurrence of Macrobrachium rosenbergii nodavirus and extra small virus like particles associated with white tail disease of m rosenbergii in india by rt pcr detection
Aquaculture, 2004Co-Authors: A Sahul S Hameed, Sri J Widada, K Yoganandhan, J R BonamiAbstract:A new disease similar to whitish tail disease (WTD) was observed in larvae and post-larvae of Macrobrachium rosenbergii in hatcheries and nursery ponds located at Nellore, Vijayawada and Chennai, India. Reverse transcriptase polymerase chain reaction (RT-PCR) assay on these samples revealed the presence of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) in infected larvae and post-larvae, as reported in China. Published primers and self-designed primers specific for MrNV were tested and compared for sensitivity with MrNV samples collected in India. Our designed primer sets 4 and 2 gave amplicons of 425 and 590 bp, respectively, with improved sensitivity to the level of 0.25 fg of total RNA when compared to 25 fg of total RNA for the previously published method. XSV detection was based on a recently published RT-PCR protocol. This is the first report on the occurrence of MrNV and XSV in WTD of freshwater prawn from India.
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characteristics of the monocistronic genome of extra small virus a virus like particle associated with Macrobrachium rosenbergii nodavirus possible candidate for a new species of satellite virus
Journal of General Virology, 2004Co-Authors: Joannes Sri Widada, J R BonamiAbstract:White tail disease (WTD) causes a high mortality rate in the freshwater prawn Macrobrachium rosenbergii. The pathogenic agent is a small virus, 25 nm in diameter, Macrobrachium rosenbergii nodavirus (MrNV), associated with extra small virus (XSV), a virus-like particle,15 nm in diameter. Sequencing of the XSV genome showed that it consists of a linear single-stranded RNA of 796 nucleotides, encoding a single structural protein, the capsid CP-17. The genome is in sense orientation, ended by a short poly(A) tail at the 3′-end. Sequence comparison did not allow XSV to be affiliated to known virus families. The hypothesis that XSV is a satellite virus, such as those described in the plant kingdom, is put forward based on its characteristics. It would constitute, therefore, the first satellite virus associated with a nodavirus.
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genome based detection methods of Macrobrachium rosenbergii nodavirus a pathogen of the giant freshwater prawn Macrobrachium rosenbergii dot blot in situ hybridization and rt pcr
Journal of Fish Diseases, 2003Co-Authors: Sri J Widada, S Durand, I Cambournac, D Qian, E Dejonghe, V Richard, J R BonamiAbstract:The availability of specific and rapid detection methods is essential for monitoring the health status of farmed species, particularly in viral diseases as in this case early diagnosis is a critical factor in containing disease outbreaks. Three complementary genome-based methods were developed for the detection of Macrobrachium rosenbergii nodavirus (MrNV), i.e. dot-blot hybridization, in situ hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR). Detection limits were established for dot-blot hybridization and RT-PCR and are c. 7 fg and 8 pg of viral RNA, respectively. In situ hybridization indicated that infection was confined to the striated muscle tissue. As a result of its sensitivity, RT-PCR can be used for in-depth investigations to examine the extent of the viral infection and establish the onset of infection in hatcheries. The application of RT-PCR on samples collected from prawn farms in China showed the possible use of this method in routine health monitoring.
Sri J Widada - One of the best experts on this subject based on the ideXlab platform.
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rapid detection of Macrobrachium rosenbergii nodavirus mrnv and extra small virus xsv the pathogenic agents of white tail disease of Macrobrachium rosenbergii de man by loop mediated isothermal amplification
Journal of Fish Diseases, 2006Co-Authors: D Pillai, J R Bonami, Sri J WidadaAbstract:: 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.
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studies on the occurrence of Macrobrachium rosenbergii nodavirus and extra small virus like particles associated with white tail disease of m rosenbergii in india by rt pcr detection
Aquaculture, 2004Co-Authors: A Sahul S Hameed, Sri J Widada, K Yoganandhan, J R BonamiAbstract:A new disease similar to whitish tail disease (WTD) was observed in larvae and post-larvae of Macrobrachium rosenbergii in hatcheries and nursery ponds located at Nellore, Vijayawada and Chennai, India. Reverse transcriptase polymerase chain reaction (RT-PCR) assay on these samples revealed the presence of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) in infected larvae and post-larvae, as reported in China. Published primers and self-designed primers specific for MrNV were tested and compared for sensitivity with MrNV samples collected in India. Our designed primer sets 4 and 2 gave amplicons of 425 and 590 bp, respectively, with improved sensitivity to the level of 0.25 fg of total RNA when compared to 25 fg of total RNA for the previously published method. XSV detection was based on a recently published RT-PCR protocol. This is the first report on the occurrence of MrNV and XSV in WTD of freshwater prawn from India.
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genome based detection methods of Macrobrachium rosenbergii nodavirus a pathogen of the giant freshwater prawn Macrobrachium rosenbergii dot blot in situ hybridization and rt pcr
Journal of Fish Diseases, 2003Co-Authors: Sri J Widada, S Durand, I Cambournac, D Qian, E Dejonghe, V Richard, J R BonamiAbstract:The availability of specific and rapid detection methods is essential for monitoring the health status of farmed species, particularly in viral diseases as in this case early diagnosis is a critical factor in containing disease outbreaks. Three complementary genome-based methods were developed for the detection of Macrobrachium rosenbergii nodavirus (MrNV), i.e. dot-blot hybridization, in situ hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR). Detection limits were established for dot-blot hybridization and RT-PCR and are c. 7 fg and 8 pg of viral RNA, respectively. In situ hybridization indicated that infection was confined to the striated muscle tissue. As a result of its sensitivity, RT-PCR can be used for in-depth investigations to examine the extent of the viral infection and establish the onset of infection in hatcheries. The application of RT-PCR on samples collected from prawn farms in China showed the possible use of this method in routine health monitoring.
D Qian - One of the best experts on this subject based on the ideXlab platform.
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genome based detection methods of Macrobrachium rosenbergii nodavirus a pathogen of the giant freshwater prawn Macrobrachium rosenbergii dot blot in situ hybridization and rt pcr
Journal of Fish Diseases, 2003Co-Authors: Sri J Widada, S Durand, I Cambournac, D Qian, E Dejonghe, V Richard, J R BonamiAbstract:The availability of specific and rapid detection methods is essential for monitoring the health status of farmed species, particularly in viral diseases as in this case early diagnosis is a critical factor in containing disease outbreaks. Three complementary genome-based methods were developed for the detection of Macrobrachium rosenbergii nodavirus (MrNV), i.e. dot-blot hybridization, in situ hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR). Detection limits were established for dot-blot hybridization and RT-PCR and are c. 7 fg and 8 pg of viral RNA, respectively. In situ hybridization indicated that infection was confined to the striated muscle tissue. As a result of its sensitivity, RT-PCR can be used for in-depth investigations to examine the extent of the viral infection and establish the onset of infection in hatcheries. The application of RT-PCR on samples collected from prawn farms in China showed the possible use of this method in routine health monitoring.
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extra small virus like particles xsv and nodavirus associated with whitish muscle disease in the giant freshwater prawn Macrobrachium rosenbergii
Journal of Fish Diseases, 2003Co-Authors: D Qian, I Cambournac, S Zhang, L Li, J R BonamiAbstract:A disease of Macrobrachium rosenbergii, the giantfreshwater prawn, farmed in China was recentlyrecorded in Zhejiang, Jiangsu, Shanghai, Guangxiand Guangdong provinces. The clinical sign of thedisease, which develops in post-larvae (PL), is awhitish appearance of the muscles, particularlynoticeable in the abdomen. Mortalities may reach100% in some hatcheries. Investigations by trans-mission electron microscopy after negative stainingof diseased PL homogenates showed the presence oftwo types of viral particles: one, unenveloped,icosahedral in shape, 26–27 nm in diameter, thesecond, much smaller, about 14–16 nm in diam-eter, designated extra small virus particle (XSV).The large virus has a genome with two pieces ofssRNA (RNA-1 and RNA-2), of 3 and 1.2 kb,respectively. Hybridization tests confirmed that thislarge virus is closely related to M. rosenbergiinodavirus (MrNV) which was isolated from dis-eased prawns in a hatchery in the French WestIndies. Its very small size and hypothesized bio-chemical and biological characteristics suggest XSVis a new type of crustacean virus. As XSV has alwaysbeen found associated with the larger virus (noda-virus) and is located in muscle and connective cellsof diseased animals, it could be an autonomousvirus, a helper-type virus or a satellite-like virus.Keywords: Macrobrachium rosenbergii, MrNV,nodavirus, RNA viruses, whitish muscle disease,extra small virus.
I Cambournac - One of the best experts on this subject based on the ideXlab platform.
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genome based detection methods of Macrobrachium rosenbergii nodavirus a pathogen of the giant freshwater prawn Macrobrachium rosenbergii dot blot in situ hybridization and rt pcr
Journal of Fish Diseases, 2003Co-Authors: Sri J Widada, S Durand, I Cambournac, D Qian, E Dejonghe, V Richard, J R BonamiAbstract:The availability of specific and rapid detection methods is essential for monitoring the health status of farmed species, particularly in viral diseases as in this case early diagnosis is a critical factor in containing disease outbreaks. Three complementary genome-based methods were developed for the detection of Macrobrachium rosenbergii nodavirus (MrNV), i.e. dot-blot hybridization, in situ hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR). Detection limits were established for dot-blot hybridization and RT-PCR and are c. 7 fg and 8 pg of viral RNA, respectively. In situ hybridization indicated that infection was confined to the striated muscle tissue. As a result of its sensitivity, RT-PCR can be used for in-depth investigations to examine the extent of the viral infection and establish the onset of infection in hatcheries. The application of RT-PCR on samples collected from prawn farms in China showed the possible use of this method in routine health monitoring.
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extra small virus like particles xsv and nodavirus associated with whitish muscle disease in the giant freshwater prawn Macrobrachium rosenbergii
Journal of Fish Diseases, 2003Co-Authors: D Qian, I Cambournac, S Zhang, L Li, J R BonamiAbstract:A disease of Macrobrachium rosenbergii, the giantfreshwater prawn, farmed in China was recentlyrecorded in Zhejiang, Jiangsu, Shanghai, Guangxiand Guangdong provinces. The clinical sign of thedisease, which develops in post-larvae (PL), is awhitish appearance of the muscles, particularlynoticeable in the abdomen. Mortalities may reach100% in some hatcheries. Investigations by trans-mission electron microscopy after negative stainingof diseased PL homogenates showed the presence oftwo types of viral particles: one, unenveloped,icosahedral in shape, 26–27 nm in diameter, thesecond, much smaller, about 14–16 nm in diam-eter, designated extra small virus particle (XSV).The large virus has a genome with two pieces ofssRNA (RNA-1 and RNA-2), of 3 and 1.2 kb,respectively. Hybridization tests confirmed that thislarge virus is closely related to M. rosenbergiinodavirus (MrNV) which was isolated from dis-eased prawns in a hatchery in the French WestIndies. Its very small size and hypothesized bio-chemical and biological characteristics suggest XSVis a new type of crustacean virus. As XSV has alwaysbeen found associated with the larger virus (noda-virus) and is located in muscle and connective cellsof diseased animals, it could be an autonomousvirus, a helper-type virus or a satellite-like virus.Keywords: Macrobrachium rosenbergii, MrNV,nodavirus, RNA viruses, whitish muscle disease,extra small virus.
Saengchan Senapin - One of the best experts on this subject based on the ideXlab platform.
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Molecular isolation and characterization of a haemocyanin of Macrobrachium rosenbergii reveal its antibacterial activities
Aquaculture Research, 2017Co-Authors: Chutima Srisuk, Saengchan Senapin, William G. Bendena, Siwaporn Longyant, Paisarn Sithigorngul, Parin ChaivisuthangkuraAbstract:Haemocyanin is a multi-subunit protein complex found in the haemolymph and is involved in the immune system of crustaceans. In this study, a haemocyanin gene of Macrobrachium rosenbergii, designated MrHc, was successfully isolated. The MrHc gene contained an open reading frame (ORF) of 1,992 nucleotides, encoding a protein of 663 amino acid residues with a molecular mass of 76.5 kDa. The deduced amino acid sequence contained distinct structural motifs of the haemocyanin superfamily, including an all-alpha domain, a copper-containing domain and an immunoglobulin-like domain. Based on the phylogenetic analysis, the MrHC protein demonstrated a close relationship with the haemocyanins of Palaemon carinicauda and Macrobrachium nipponense. The MrHc gene was expressed in various shrimp tissues, including the hepatopancreas, gill, haemocytes, stomach and muscle. After Macrobrachium rosenbergii nodavirus (MrNV) challenge tests, the MrHc gene was up-regulated 237-fold at day 2. A recombinant protein of the MrHc immunoglobulin-like domain exhibited antibacterial activity against Vibrio vulnificus, V. parahaemolyticus, Aeromonas caviae, A. veronii, A. hydrophila and Bacillus cereus. This study suggested that MrHc may play important roles in the shrimp innate immune response to MrNV infection and bacterial infection.
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infections of mrnv Macrobrachium rosenbergii nodavirus in cultivated whiteleg shrimp penaeus vannamei in asia
Aquaculture, 2012Co-Authors: Saengchan Senapin, Chatlada Jaengsanong, Kornsunee Phiwsaiya, Satit Prasertsri, Kesinee Laisutisan, Niti Chuchird, Chalor LimsuwanAbstract:Abstract Macrobrachium rosenbergii nodavirus ( Mr NV) is well-known as a major pathogen that causes whitened muscles and high mortality in the freshwater prawn Macrobrachium rosenbergii . Recently, it has also been reported to cause white muscles and high mortality in postlarvae of the marine shrimp Penaeus ( Penaeus ) monodon and Penaeus ( Fenneropenaeus ) indicus in India. The latter report stimulated us to re-examine specimens from Asian shrimp farms that had experienced high mortality in Penaeus ( Litopenaeus ) vannamei with white muscles but tested negative for infectious myonecrosis virus (IMNV) and Penaeus vannamei nodavirus ( Pv NV) by RT-PCR analysis. Some of these specimens were indeed positive for Mr NV by RT-PCR. Sequences of the two single-stranded RNA fragments in the Mr NV genome were amplified from these P. vannamei specimens and found to share ~ 97% nucleic acid sequence identity with the Mr NV sequences deposited at GenBank ( NC_005094 and NC_005095 NC_005094 NC_005095 ). Extra small virus (XSV) usually associated with Mr NV, was detected in some but not all of the samples. Infectivity tests were performed by feeding P. vannamei with minced tissues from Mr NV-infected M. rosenbergii . The assays were preformed at low salinity of 2 ppt and at two different water temperatures of approximately 22 °C and 28 °C. It was revealed that shrimp exhibited a higher infectivity and mortality at the lower temperature. Our findings suggested that P. vannamei is an additional species that is susceptible to Mr NV and that low temperature together with low salinity of rearing water may increase the severity of infections leading to significant mortality.
