The Experts below are selected from a list of 21531 Experts worldwide ranked by ideXlab platform
Shaoping Weng - One of the best experts on this subject based on the ideXlab platform.
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Development of a gene-deleted live attenuated candidate vaccine against fish virus (ISKNV) with low pathogenicity and high protection
'Elsevier BV', 2021Co-Authors: Ruoyun Zeng, Shaoping Weng, Yifan Lin, Zhiyong Luo, Weiqiang Pan, Chang-jun GuoAbstract:Summary: Aquaculture provides important food, nutrition, and income sources for humans. However, aquaculture industry is seriously threatened by viral diseases. Infectious spleen and Kidney Necrosis virus (ISKNV) disease causes high mortality and economic losses to the fish culture industry in Asia and has been listed as a certifiable disease by the International Epizootic Office. Vaccine development is urgent to control this disease. Here, a gene-deleted live attenuated candidate vaccine (ΔORF022L) against ISKNV with low pathogenicity and high protection was developed. ΔORF022L replicated well in mandarin fish fry-1 cells and showed similar structure with wild-type ISKNV. However, the pathogenicity was significantly lower as 98% of the mandarin fish infected with ΔORF022L survived, whereas all those infected with wild-type ISKNV died. Of importance, 100% of the ΔORF022L-infected fish survived the ISKNV challenge. ΔORF022L induced anti-ISKNV specific antibody response and upregulation of immune-related genes. This work could be beneficial to the control of fish diseases
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transcriptional programs of infectious spleen and Kidney Necrosis virus isknv in vitro and in vivo
Virus Genes, 2020Co-Authors: Lichao Huang, Zhixun Guo, Shaoping WengAbstract:Infectious spleen and Kidney Necrosis virus (ISKNV), causing serious infectious diseases to marine and freshwater fishes, is the type species of the genus Megalocytivirus, family Iridoviridae. In this study, the transcriptional programs of ISKNV in vitro (MFF-1 cells) and in vivo (spleens from mandarin fish) were investigated using real-time PCR. Transcription of all the putative open reading frames (ORFs) of ISKNV was verified. The temporal expression patterns of ISKNV ORFs in vitro and in vivo, including peak expression times (PETs) and relative maximal expression levels, were determined and compared. The K-means clustering with Spearman rank correlation was generated in heat maps constructed based on ISKNV ORF expression profiles in vivo and in vitro. The current study may provide a global picture of ISKNV infection at the transcription level and help better understand the molecular pathogenic mechanism of megalocytiviruses.
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Identification of infectious spleen and Kidney Necrosis virus (ISKNV)-encoded microRNAs.
Virus genes, 2020Co-Authors: Qiong Xia, Shaoping WengAbstract:MicroRNAs (miRNAs) are small noncoding RNAs that post-transcriptionally regulate gene expression by complementary binding to target mRNAs. Virus-encoded miRNAs play important roles in virus life cycle and virus-host interactions. Viruses from the Megalocytivirus genus, family Iridoviridae, infect a wide range of fishes, bringing great challenges to aquaculture. Infectious spleen and Kidney Necrosis virus (ISKNV) is the type species of the Megalocytivirus genus. In this study, using Illumina sequencing coupled with miRNA precursor prediction and stem-loop real-time PCR, 14 putative ISKNV-encoded miRNAs were preliminarily identified from ISKNV-infected mandarin fish MFF-1 cells. To initially study their functions, inhibitors of the 14 viral miRNAs were synthesized and transfected into MFF-1 cells, which were further infected with ISKNV. The results showed that these viral miRNAs could affect the virus titers in the supernatant of ISKNV-infected cells and the expression of major capsid protein (MCP). Moreover, we observed that inhibition of several ISKNV miRNAs had different effects on MCP expression and on titer of released virus, suggesting complex roles of viral miRNAs in ISKNV infection. The current study may provide a fundamental information for further identification and functional studies on miRNAs encoded by Megalocytivirus.
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the roles of mandarin fish sting in innate immune defense against infectious spleen and Kidney Necrosis virus infections
Fish & Shellfish Immunology, 2020Co-Authors: Jian He, Shaoping Weng, Zhimin Li, Yang Yu, Jianguo HeAbstract:Abstract The mandarin fish Siniperca chuatsi is a cultured freshwater fish species that is popular in China because of its high market value. With the development of high-density cultural mode in mandarin fish, viral diseases such as Infectious spleen and Kidney Necrosis virus (ISKNV) are becoming increasingly serious. Stimulator of interferon genes (STING) is a central component in the innate immune response to cytosolic DNA and RNA derived from different pathogens. However, the roles of STING in innate immune response of mandarin fish remain unknown. In the present study, S. chuatsi STING (scSTING)-mediated host immune response against ISKNV infection was investigated. ScSTING transcription level increased remarkably in response to ISKNV infection, LPS, PMA, or poly (I:C) stimulation in mandarin fish fry (MFF-1) cells. Immunofluorescence results showed that scSTING localized majorly in the endoplasmic reticulum. scSTING overexpression remarkably increased the expression levels of scIFN-h, scMx, scISG15, scPKR, scViperin, scIL-1β, scIL-18, and scTNF-α genes. IFN-β-luciferase report assay results showed that the relative expressions of luciferin were remarkably increased in MFF-1 cells. Site mutation of serine (S) on C-terminus of scSTING showed that both S388 and S396 were important for mediated signaling. Furthermore, scSTING overexpression inhibited ISKNV infection, and knockdown of scSTING promoted ISKNV infection, indicating that scSTING could suppress ISKNV infection in MFF-1 cells. These observations suggested that the scSTING played an important role in innate immune against ISKNV infection. Our work would help elucidate the roles of teleost fish STING in innate immunity.
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deletion of the infectious spleen and Kidney Necrosis virus orf069l reduces virulence to mandarin fish siniperca chuatsi
Fish & Shellfish Immunology, 2019Co-Authors: Yifan Lin, Shaoping Weng, Ruoyun Zeng, Zhiyong Luo, Weiqiang Pan, Chang-jun GuoAbstract:Mandarin fish (Siniperca chuatsi) is a significant cultured species with high added value in China. With the expansion of farming, diseases of mandarin fish such as Infectious spleen and Kidney Necrosis virus (ISKNV) diseases are becoming more and more serious. Human endogenous retrovirus subfamily H long terminal repeat associating protein 2 (HHLA2) is a type 1 transmembrane molecule with three extracellular Ig domains (IgV-IgC-IgV) and plays important roles in the T cell proliferation and tumorigenesis. The HHLA2-homologues have not been found in virus. In this study, a viral HHLA2 protein encoded by ISKNV ORF069L was identified and the virulence of the deleted ORF069L reconstruction ISKNV strain (ΔORF069L) was investigated. ISKNV ORF069L gene was predicted to encode a 222-amino acids peptide. The bioinformation analysis revealed that ISKNV ORF069L contained an Ig HHLA2 domain and was homologous to vertebrate B7-CD28 family proteins. The recombinant virus strain of ΔORF069L was constructed by homologous recombination technology. The virus titer and growth curves between ISKNV wild type (WT) and ΔORF069L on cellular level showed no significant differences indicating that the ORF069L did not influence the ISKNV replication. The expression levels of immune-related genes (Mx1, IL-1β, IL-8, TNF-a and IgM) were increased in fish infected with ΔORF069L, compared to those in fish infected with ISKNV WT. Furthermore, the lethality caused by ΔORF069L declined by 40% compared with ISKNV WT, indicating that ORF069L was a virulence gene of ISKNV. Most importantly, the protection rate was nearly 100% for fish immunized with ΔORF069L strain. Those results suggested that ΔORF069L could be developed as a potential attenuated vaccine against ISKNV. Our work will be beneficial to promote the development of gene deletion attenuated vaccines for ISKNV disease.
Qiang Lin - One of the best experts on this subject based on the ideXlab platform.
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gefitinib inhibits infectious spleen and Kidney Necrosis virus infection in vivo and vitro by blocking virus endocytosis
Aquaculture, 2021Co-Authors: Yinjie Niu, Lihui Liu, Qiang Lin, Xia Luo, Hongru LiangAbstract:Abstract Infectious spleen and Kidney Necrosis virus disease (ISKNVD) caused significant economic losses to the fishery industry. So it is urgent to develop an effective measure for prevention ISKNVD. In this study, we investigated the effects of gefitinib against ISKNV in vitro and vivo. The gefitinib inhibited viral protein synthesis and decreased viral titers with the dose-dependent manner in Chinese perch brain cells (CPB) cells. Gefitinib was observed to inhibit ISKNV infection by blocking ISKNV entry into CPB cells. Interestingly, gefitinib inhibited EGFR/PI3K phosphorylation and suppressed microfilament gathering induced by ISKNV. At the same time, gefitinib inhibited ISKNV infection and decreased pathogenicity of ISKNV in vivo. These results suggested that gefitinib prevented ISKNV infection by blocking virus-mediated endocytosis. This research provided potential of gefitinib for preventing ISKNVD.
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development and application of a sensitive droplet digital pcr ddpcr for the detection of infectious spleen and Kidney Necrosis virus
Aquaculture, 2020Co-Authors: Qiang Lin, Lihui Liu, Hongru Liang, Yinjie Niu, Yuanya Wen, Zhibin HuangAbstract:Abstract Infectious spleen and Kidney Necrosis, caused by the infectious spleen and Kidney Necrosis virus (ISKNV), has damaged the economy of the world fish industry. Droplet digital PCR (ddPCR) is a novel, sensitive, accurate and absolute quantitation method that does not require a standard curve. In this study, we established a sensitive ddPCR method to rapidly detect and quantify ISKNV DNA. The established method is highly specific to ISKNV and does not cross-react with other iridoviruses. The detection limit of the ddPCR was found to be 1.5 copies/μL, which is much lower than the 34 copies/μL determined for TaqMan real-time PCR (qPCR). This indicated that the sensitivity of the ddPCR assay was 20-fold higher than the sensitivity of the qPCR assay. We explored the feasibility of ddPCR to detect ISKNV from 23 fry samples (mandarin fish, Siniperca chuatsi) and compared the data with qPCR, in terms of sensitivity and accuracy. The detection results for fry samples showed that the positive detection rate of ddPCR (65.22%) was higher than that of qPCR (30.43%). In conclusion, the ddPCR method shows superiority for detection in samples with low ISKNV viral loads, which will facilitate the surveillance of sources and transmission routes of ISKNV.
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single walled carbon nanotubes as delivery vehicles enhance the immunoprotective effect of an immersion dna vaccine against infectious spleen and Kidney Necrosis virus in mandarin fish
Fish & Shellfish Immunology, 2020Co-Authors: Zhao Zhao, Gao Xue Wang, Qiang Lin, Chen Zhang, Zhibin Huang, Mi Zhao, Bin ZhuAbstract:As a high mortality disease, Infectious spleen and Kidney Necrosis virus (ISKNV) can cause massive economic damage on mandarin fish farming industry in China, which seriously hindered the development of mandarin fish farming industry. In this research, SWCNTs (single-walled carbon nanotubes) as a candidate for DNA vaccine carrier was vaccinated by immersion (1, 2, 5, 10, 20 mg/L) in juvenile mandarin fish. In muscle, spleen and Kidney tissues, the results showed that transcription and expression of MCP gene can be detected in pcDNA-MCP and SWCNTs-pcDNA-MCP groups after bath immunization. The immune response (immune-related genes expression, serum antibody production, enzyme activities and C3 content) was significantly enhanced in fish which vaccinated with SWCNTs-pcDNA-MCP in comparison with those vaccinated with pcDNA-MCP alone. After 14 d challenge, the RPS (relative percentage survival) can be enhanced which using SWCNTs as a carrier in SWCNTs-pcDNA-MCP (82.4%) group at 20 mg/L (the highest vaccine dose) than the naked pcDNA-MCP (54.2%) group. This study reveals that functionalized SWCNTs could be a promising immersion DNA vaccine carrier in aquaculture.
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accelerated metabolite levels of aerobic glycolysis and the pentose phosphate pathway are required for efficient replication of infectious spleen and Kidney Necrosis virus in chinese perch brain cells
Biomolecules, 2019Co-Authors: Xixi Guo, Lihui Liu, Qiang Lin, Hongru Liang, Yinjie Niu, Zhibin HuangAbstract:Glucose is a main carbon and energy source for virus proliferation and is usually involved in the glycolysis, pentose phosphate pathway (PPP), and tricarboxylic acid cycle (TCA cycle) pathways. In this study, we investigated the roles of glucose-related metabolic pathways during the replication of infectious spleen and Kidney Necrosis virus (ISKNV), which has caused serious economic losses in the cultured Chinese perch (Siniperca chuatsi) industry. We found that ISKNV infection enhanced the metabolic pathways of the PPP and the TCA cycle at the early stage of the ISKNV infection cycle and enhanced the glycolysis pathway at the late stage of the ISKNV infection cycle though the comprehensive analysis of transcriptomics, proteomics, and metabolomics. The advanced results proved that ISKNV replication induced upregulation of aerobic glycolysis at the late stage of ISKNV infection cycle and aerobic glycolysis were required for ISKNV multiplication. In addition, the PPP, providing nucleotide biosynthesis, was also required for ISKNV multiplication. However, the TCA cycle involving glucose was not important and necessary for ISKNV multiplication. The results reported here provide new insights into viral pathogenesis mechanism of metabolic shift, as well as antiviral treatment strategies.
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immersion vaccination of mandarin fish siniperca chuatsi against infectious spleen and Kidney Necrosis virus with a swcnts based subunit vaccine
Fish & Shellfish Immunology, 2019Co-Authors: Zhao Zhao, Gao Xue Wang, Qiang Lin, Chen Zhang, Zhibin Huang, Yijun Jia, Dekui Qiu, Bin ZhuAbstract:Infectious spleen and Kidney Necrosis virus (ISKNV) cause a high mortality disease which lead to significant economic loss on mandarin fish in China. There is no effective drug or vaccine against this fatal disease at present. Meanwhile, many drugs and vaccines had no effect in many cases account of several impenetrable barriers (cell, skin and gastrointestinal tract). Here we reported an immersion subunit vaccine system (SWCNTs-MCP) encoding MCP gene of ISKNV based on single-walled carbon nanotubes (SWCNTs). To evaluate its efficacy against ISKNV, we found a stronger and longer duration immune response (serum antibody production, enzyme activities and immune-related genes expression) can be induced in fish vaccinated with SWCNTs-MCP in comparison with those vaccinated with MCP alone. Importantly, SWCNTs can increase the immune protective effect of naked subunit vaccine by ca. 23.8%. Thereby, this study demonstrates that SWCNTs as a promising carrier for subunit vaccine might be used to vaccinate large-scale juvenile mandarin fish by bath administration approach.
Zhibin Huang - One of the best experts on this subject based on the ideXlab platform.
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development and application of a sensitive droplet digital pcr ddpcr for the detection of infectious spleen and Kidney Necrosis virus
Aquaculture, 2020Co-Authors: Qiang Lin, Lihui Liu, Hongru Liang, Yinjie Niu, Yuanya Wen, Zhibin HuangAbstract:Abstract Infectious spleen and Kidney Necrosis, caused by the infectious spleen and Kidney Necrosis virus (ISKNV), has damaged the economy of the world fish industry. Droplet digital PCR (ddPCR) is a novel, sensitive, accurate and absolute quantitation method that does not require a standard curve. In this study, we established a sensitive ddPCR method to rapidly detect and quantify ISKNV DNA. The established method is highly specific to ISKNV and does not cross-react with other iridoviruses. The detection limit of the ddPCR was found to be 1.5 copies/μL, which is much lower than the 34 copies/μL determined for TaqMan real-time PCR (qPCR). This indicated that the sensitivity of the ddPCR assay was 20-fold higher than the sensitivity of the qPCR assay. We explored the feasibility of ddPCR to detect ISKNV from 23 fry samples (mandarin fish, Siniperca chuatsi) and compared the data with qPCR, in terms of sensitivity and accuracy. The detection results for fry samples showed that the positive detection rate of ddPCR (65.22%) was higher than that of qPCR (30.43%). In conclusion, the ddPCR method shows superiority for detection in samples with low ISKNV viral loads, which will facilitate the surveillance of sources and transmission routes of ISKNV.
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single walled carbon nanotubes as delivery vehicles enhance the immunoprotective effect of an immersion dna vaccine against infectious spleen and Kidney Necrosis virus in mandarin fish
Fish & Shellfish Immunology, 2020Co-Authors: Zhao Zhao, Gao Xue Wang, Qiang Lin, Chen Zhang, Zhibin Huang, Mi Zhao, Bin ZhuAbstract:As a high mortality disease, Infectious spleen and Kidney Necrosis virus (ISKNV) can cause massive economic damage on mandarin fish farming industry in China, which seriously hindered the development of mandarin fish farming industry. In this research, SWCNTs (single-walled carbon nanotubes) as a candidate for DNA vaccine carrier was vaccinated by immersion (1, 2, 5, 10, 20 mg/L) in juvenile mandarin fish. In muscle, spleen and Kidney tissues, the results showed that transcription and expression of MCP gene can be detected in pcDNA-MCP and SWCNTs-pcDNA-MCP groups after bath immunization. The immune response (immune-related genes expression, serum antibody production, enzyme activities and C3 content) was significantly enhanced in fish which vaccinated with SWCNTs-pcDNA-MCP in comparison with those vaccinated with pcDNA-MCP alone. After 14 d challenge, the RPS (relative percentage survival) can be enhanced which using SWCNTs as a carrier in SWCNTs-pcDNA-MCP (82.4%) group at 20 mg/L (the highest vaccine dose) than the naked pcDNA-MCP (54.2%) group. This study reveals that functionalized SWCNTs could be a promising immersion DNA vaccine carrier in aquaculture.
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accelerated metabolite levels of aerobic glycolysis and the pentose phosphate pathway are required for efficient replication of infectious spleen and Kidney Necrosis virus in chinese perch brain cells
Biomolecules, 2019Co-Authors: Xixi Guo, Lihui Liu, Qiang Lin, Hongru Liang, Yinjie Niu, Zhibin HuangAbstract:Glucose is a main carbon and energy source for virus proliferation and is usually involved in the glycolysis, pentose phosphate pathway (PPP), and tricarboxylic acid cycle (TCA cycle) pathways. In this study, we investigated the roles of glucose-related metabolic pathways during the replication of infectious spleen and Kidney Necrosis virus (ISKNV), which has caused serious economic losses in the cultured Chinese perch (Siniperca chuatsi) industry. We found that ISKNV infection enhanced the metabolic pathways of the PPP and the TCA cycle at the early stage of the ISKNV infection cycle and enhanced the glycolysis pathway at the late stage of the ISKNV infection cycle though the comprehensive analysis of transcriptomics, proteomics, and metabolomics. The advanced results proved that ISKNV replication induced upregulation of aerobic glycolysis at the late stage of ISKNV infection cycle and aerobic glycolysis were required for ISKNV multiplication. In addition, the PPP, providing nucleotide biosynthesis, was also required for ISKNV multiplication. However, the TCA cycle involving glucose was not important and necessary for ISKNV multiplication. The results reported here provide new insights into viral pathogenesis mechanism of metabolic shift, as well as antiviral treatment strategies.
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immersion vaccination of mandarin fish siniperca chuatsi against infectious spleen and Kidney Necrosis virus with a swcnts based subunit vaccine
Fish & Shellfish Immunology, 2019Co-Authors: Zhao Zhao, Gao Xue Wang, Qiang Lin, Chen Zhang, Zhibin Huang, Yijun Jia, Dekui Qiu, Bin ZhuAbstract:Infectious spleen and Kidney Necrosis virus (ISKNV) cause a high mortality disease which lead to significant economic loss on mandarin fish in China. There is no effective drug or vaccine against this fatal disease at present. Meanwhile, many drugs and vaccines had no effect in many cases account of several impenetrable barriers (cell, skin and gastrointestinal tract). Here we reported an immersion subunit vaccine system (SWCNTs-MCP) encoding MCP gene of ISKNV based on single-walled carbon nanotubes (SWCNTs). To evaluate its efficacy against ISKNV, we found a stronger and longer duration immune response (serum antibody production, enzyme activities and immune-related genes expression) can be induced in fish vaccinated with SWCNTs-MCP in comparison with those vaccinated with MCP alone. Importantly, SWCNTs can increase the immune protective effect of naked subunit vaccine by ca. 23.8%. Thereby, this study demonstrates that SWCNTs as a promising carrier for subunit vaccine might be used to vaccinate large-scale juvenile mandarin fish by bath administration approach.
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co infections of infectious spleen and Kidney Necrosis virus and siniperca chuatsi rhabdovirus in chinese perch siniperca chuatsi
Microbial Pathogenesis, 2017Co-Authors: Qiang Lin, Zhibin Huang, Quanyuan Wan, Wenjie Chen, Yunmao Huang, Lijuan Zhao, Li LinAbstract:In spite of the quite common co-infections of viruses in the cultured fish, most of the previous studies have just simply focused on the infection of a single pathogen. In this report, we observed that about 13% of cultured Chinese perch have been co-infected by infectious spleen and Kidney Necrosis virus (ISKNV) and Siniperca chuatsi rhabdovirus (SCRV). Furthermore, Chinese perch could co-infected by ISKNV and SCRV by intraperitoneally injection with the two viruses. Interestingly, we revealed that the two viruses could even co-infect a single cell of Chinese perch in vivo and a single Chinese perch brain cells (CPB) cell in vitro. The dynamic co-infected viruses loads in the different tissues of Chinese perch showed dependent. When CPB cells were infected with the same 10 MOI of SCRV and ISKNV, the replication of SCRV overwhelmed the replication of ISKNV. When the MOI of ISKNV (10 MOI) was 10,000 times of MOI of SCRV (0.001 MOI), the dynamic virus loads of the two viruses in CPB cells indicated that co-infections could synergistically stimulate both viruses replication at the late time points but not at early time points. The co-infections of ISKNV and SCRV in the cultured Chinese perch will shed a new light on the prevention of the viral diseases of Chinese perch. The development of multivalent vaccine which could be effective for preventing against the co-infections of the viruses is highly needed.
Lihui Liu - One of the best experts on this subject based on the ideXlab platform.
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gefitinib inhibits infectious spleen and Kidney Necrosis virus infection in vivo and vitro by blocking virus endocytosis
Aquaculture, 2021Co-Authors: Yinjie Niu, Lihui Liu, Qiang Lin, Xia Luo, Hongru LiangAbstract:Abstract Infectious spleen and Kidney Necrosis virus disease (ISKNVD) caused significant economic losses to the fishery industry. So it is urgent to develop an effective measure for prevention ISKNVD. In this study, we investigated the effects of gefitinib against ISKNV in vitro and vivo. The gefitinib inhibited viral protein synthesis and decreased viral titers with the dose-dependent manner in Chinese perch brain cells (CPB) cells. Gefitinib was observed to inhibit ISKNV infection by blocking ISKNV entry into CPB cells. Interestingly, gefitinib inhibited EGFR/PI3K phosphorylation and suppressed microfilament gathering induced by ISKNV. At the same time, gefitinib inhibited ISKNV infection and decreased pathogenicity of ISKNV in vivo. These results suggested that gefitinib prevented ISKNV infection by blocking virus-mediated endocytosis. This research provided potential of gefitinib for preventing ISKNVD.
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development and application of a sensitive droplet digital pcr ddpcr for the detection of infectious spleen and Kidney Necrosis virus
Aquaculture, 2020Co-Authors: Qiang Lin, Lihui Liu, Hongru Liang, Yinjie Niu, Yuanya Wen, Zhibin HuangAbstract:Abstract Infectious spleen and Kidney Necrosis, caused by the infectious spleen and Kidney Necrosis virus (ISKNV), has damaged the economy of the world fish industry. Droplet digital PCR (ddPCR) is a novel, sensitive, accurate and absolute quantitation method that does not require a standard curve. In this study, we established a sensitive ddPCR method to rapidly detect and quantify ISKNV DNA. The established method is highly specific to ISKNV and does not cross-react with other iridoviruses. The detection limit of the ddPCR was found to be 1.5 copies/μL, which is much lower than the 34 copies/μL determined for TaqMan real-time PCR (qPCR). This indicated that the sensitivity of the ddPCR assay was 20-fold higher than the sensitivity of the qPCR assay. We explored the feasibility of ddPCR to detect ISKNV from 23 fry samples (mandarin fish, Siniperca chuatsi) and compared the data with qPCR, in terms of sensitivity and accuracy. The detection results for fry samples showed that the positive detection rate of ddPCR (65.22%) was higher than that of qPCR (30.43%). In conclusion, the ddPCR method shows superiority for detection in samples with low ISKNV viral loads, which will facilitate the surveillance of sources and transmission routes of ISKNV.
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accelerated metabolite levels of aerobic glycolysis and the pentose phosphate pathway are required for efficient replication of infectious spleen and Kidney Necrosis virus in chinese perch brain cells
Biomolecules, 2019Co-Authors: Xixi Guo, Lihui Liu, Qiang Lin, Hongru Liang, Yinjie Niu, Zhibin HuangAbstract:Glucose is a main carbon and energy source for virus proliferation and is usually involved in the glycolysis, pentose phosphate pathway (PPP), and tricarboxylic acid cycle (TCA cycle) pathways. In this study, we investigated the roles of glucose-related metabolic pathways during the replication of infectious spleen and Kidney Necrosis virus (ISKNV), which has caused serious economic losses in the cultured Chinese perch (Siniperca chuatsi) industry. We found that ISKNV infection enhanced the metabolic pathways of the PPP and the TCA cycle at the early stage of the ISKNV infection cycle and enhanced the glycolysis pathway at the late stage of the ISKNV infection cycle though the comprehensive analysis of transcriptomics, proteomics, and metabolomics. The advanced results proved that ISKNV replication induced upregulation of aerobic glycolysis at the late stage of ISKNV infection cycle and aerobic glycolysis were required for ISKNV multiplication. In addition, the PPP, providing nucleotide biosynthesis, was also required for ISKNV multiplication. However, the TCA cycle involving glucose was not important and necessary for ISKNV multiplication. The results reported here provide new insights into viral pathogenesis mechanism of metabolic shift, as well as antiviral treatment strategies.
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First report of megalocytivirus (iridoviridae) in cultured bluegill sunfish, Lepomis macrochirus, in China.
Microbial pathogenesis, 2019Co-Authors: Lihui Liu, Qiang Lin, Hongru Liang, Yinjie NiuAbstract:The bluegill sunfish, Lepomis macrochirus, is an important aquacultural and recreational species in southern China because of its excellent taste, rapid growth rate, and good looks. At present, few pathogens are known to affect the bluegill sunfish. However, an iridovirus-like disease recently caused heavy losses to the bluegill sunfish aquaculture industry in Guangdong, China. We report that a virus, designated BSMIV-SD-20171020, was isolated from diseased bluegill sunfish in China. The isolate was efficiently propagated in a Chinese perch brain (CPB) cell line. The cytopathic effect was observed, the MCP gene PCR amplified, and the virus observed with electron microscopy. Its viral titer in CPB cells reached 104.13 TCID50 mL-1. The mortality rate was 100% when bluegill sunfish were challenged with BSMIV-SD-20171020 at a dose of 103.13 TCID50/fish. A histopathological examination revealed basophilic hypertrophied cells in the intestine, liver, and spleen. A nucleotide sequence alignment and phylogenetic analysis of the major capsid protein revealed that isolate BSMIV-SD-20171020 is the species Infectious spleen and Kidney Necrosis virus (ISKNV), in the genus Megalocytivirus.
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application and development of a taqman real time pcr for detecting infectious spleen and Kidney Necrosis virus in siniperca chuatsi
Microbial Pathogenesis, 2017Co-Authors: Qiang Lin, Lihui Liu, Hongru Liang, Huizhi Guo, Shuwen Yin, Venkatesh Kumaresan, Zhibin HuangAbstract:Infectious spleen and Kidney Necrosis virus (ISKNV) is one of the major epidemiological agents that had caused great economic loss in Chinese perch (Siniperca chuatsi). In this study, a specific TaqMan real-time PCR was developed using a pair of primers and a TaqMan probe specific to the ORF007 gene of ISKNV to rapidly detect ISKNV copies in Chinese perch samples. This assay was optimized to produce linearity from 8.75 × 108 to 8.75 × 101 copies in standard curve with an efficiency of 98% and a R2 value of 0.9999. Moreover, the minimum detection limit of this assay was 10,000 times more sensitive than that of conventional PCR method. The coefficients of variation of intra- and inter-assay repeatability were less than 2.4% and 3.3%, respectively. The viral distribution in different tissues of diseased Chinese perch was evaluated by TaqMan real-time PCR method and the highest level of viral copies was detected in spleen. Among the 76 diseased Chinese perch clinical samples, 35 and 29 were positive samples based on the TaqMan real-time PCR and conventional PCR methods, respectively, indicating that the TaqMan real-time PCR was more sensitive than conventional PCR. Therefore, the TaqMan real-time PCR should be a useful tool for the early surveillance and quantitation of ISKNV.
Bin Zhu - One of the best experts on this subject based on the ideXlab platform.
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Single-walled carbon nanotubes as drug carrier loaded with moroxydine hydrochloride against infectious spleen and Kidney Necrosis virus in mandarin fish
Aquaculture, 2021Co-Authors: Zhao Zhao, Gao Xue Wang, Zi-rao Guo, Chen Zhang, Yan Xiong, Bin ZhuAbstract:Abstract Infectious diseases of viral origin cause major aquatic production losses in different parts of the world. There are lots of limited therapeutic effects for large amounts of antiviral drugs because of several impenetrable barriers (cell, skin, and gastrointestinal tract). In this study, we selected moroxydine hydrochloride (Mor) for the treatment of infectious spleen and Kidney Necrosis virus (ISKNV) and choose single-walled carbon nanotubes (SWCNTs) as drug carrier by immersion bath on mandarin fish infected with ISKNV. The results showed that increasing Mor intake was observed by SWCNTs carrier and therapeutic dosage to kill ISKNV is significantly reduced. After 4 h treatment, the concentration of Mor was 103.48 μg/g in 40 mg/L Mor group, and 182.35 μg/g in 40 mg/L Mor-SWCNTs groups. Comparing with the same dosage in Mor group, the detention time of Mor was extended more than 48 h in Mor-SWCNTs group. After 11 d treatment, cumulative mortality was 11.51% and infection rate was 3.81% in 40 mg/L Mor-SWCNTs group. In the same dosage of Mor group, cumulative mortality was 43.34% and infection rate was 22.67%. Moreover, enzyme activities, complement (C3) content and immune-related genes expression were significantly increased in Mor and Mor-SWCNTs groups than control group, herein all these mentioned activities reached the highest level at 3 days post-treatment. Our results indicated that drug delivery with functionalized SWCNTs can improve the antiviral effect on mandarin fish and has a potential application value to control fish viral diseases in aquaculture.
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in vivo efficacy of ganciclovir by using single walled carbon nanotubes as a drug carrier against infectious spleen and Kidney Necrosis virus in mandarin fish
Aquaculture, 2020Co-Authors: Zhao Zhao, Gao Xue Wang, Chen Zhang, Yijun Jia, Bin ZhuAbstract:Abstract Infectious spleen and Kidney Necrosis virus (ISKNV) cause a high mortality disease which leads to a significant economic loss on mandarin fish in China. At present, there is no effective drug against this fatal disease because of several impenetrable barriers (cell, skin and gastrointestinal tract). In this study, we selected ganciclovir for the treatment of ISKNV and choose single-walled carbon nanotubes (SWCNTs) as ganciclovir carrier by immersion bath on mandarin fish infected with ISKNV. After 7 d post-infection, cumulative mortality was 14.75% and infection rate was 26.55% in naked ganciclovir group (40 mg/L). Meanwhile, cumulative mortality of 32.50% and infection rate of 15.30% were observed in G-SWCNTs group (20 mg/L). In the control group, cumulative mortality was 88.75% and the infection rate was 100%. At 6 h post-infection, the concentration of ganciclovir was detected to be 34.58 μg/g in naked ganciclovir group (20 mg/L). And in G-SWCNTs groups (10 mg/L) at 6 h, the concentration of ganciclovir was 39.35 μg/g. Besides, the time detention of ganciclovir in G-SWCNTs group was significantly extended compared with the naked ganciclovir group in the same dosage. Moreover, enzyme activities, complement (C3) content and immune-related genes expression were used to verify the enhancement effect induced in G-SWCNTs groups, herein all these mentioned activities reached the highest level at 3 days post-treatment. The results showed that ganciclovir-SWCNTs can be a plausible candidate for preventing and controlling ISKNV among farmed mandarin fish.
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optimization of the efficacy of a swcnts based subunit vaccine against infectious spleen and Kidney Necrosis virus in mandarin fish
Fish & Shellfish Immunology, 2020Co-Authors: Zhao Zhao, Gao Xue Wang, Chen Zhang, Yan Xiong, Yijun Jia, Dekui Qiu, Bin ZhuAbstract:Abstract Infectious spleen and Kidney Necrosis virus (ISKNV) cause a high mortality disease which brings substantial economic losses to the mandarin fish culture industry in China. This study was aimed at optimizing the efficacy of a SWCNTs-based immersion subunit vaccine (SWCNTs-M-MCP) which as a promising vaccine against ISKNV. Mandarin fish were vaccinated by immersion, then we designed an orthogonal experiment to optimize different parameters affecting vaccination such as immune duration of bath immunization, immune dose, and fish density when immunized. Our results showed that the highest relative percent survival (86.7%) was found in the group 6 with 8 h of immune duration, 20 mg/L of immune dose, and 8 fish per liter of fish density. And other immune responses (serum antibody production, enzyme activities, and immune-related genes expression) also demonstrated similar results. In addition, the expression of IRF-I in group 6 (8 h, 20 mg/L, 8 fish per liter) was significant extents, and about 16-folds increases were obtained than the control group at 21 d post-vaccination. And the highest specific antibody response was significantly increased (more than 4-folds) than control group which was found in group 6. The optimum immune duration, immune dose, and fish density of SWCNTs-M-MCP were 8 h, 20 mg/L, 8 fish per liter, respectively. Importantly, our results also showed that immune duration had the greatest effect on the immune response of our vaccine, followed by immune dose. The study reported herein provides a helpful reference for the effective use of vaccine in fish farming industry.
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single walled carbon nanotubes as delivery vehicles enhance the immunoprotective effect of an immersion dna vaccine against infectious spleen and Kidney Necrosis virus in mandarin fish
Fish & Shellfish Immunology, 2020Co-Authors: Zhao Zhao, Gao Xue Wang, Qiang Lin, Chen Zhang, Zhibin Huang, Mi Zhao, Bin ZhuAbstract:As a high mortality disease, Infectious spleen and Kidney Necrosis virus (ISKNV) can cause massive economic damage on mandarin fish farming industry in China, which seriously hindered the development of mandarin fish farming industry. In this research, SWCNTs (single-walled carbon nanotubes) as a candidate for DNA vaccine carrier was vaccinated by immersion (1, 2, 5, 10, 20 mg/L) in juvenile mandarin fish. In muscle, spleen and Kidney tissues, the results showed that transcription and expression of MCP gene can be detected in pcDNA-MCP and SWCNTs-pcDNA-MCP groups after bath immunization. The immune response (immune-related genes expression, serum antibody production, enzyme activities and C3 content) was significantly enhanced in fish which vaccinated with SWCNTs-pcDNA-MCP in comparison with those vaccinated with pcDNA-MCP alone. After 14 d challenge, the RPS (relative percentage survival) can be enhanced which using SWCNTs as a carrier in SWCNTs-pcDNA-MCP (82.4%) group at 20 mg/L (the highest vaccine dose) than the naked pcDNA-MCP (54.2%) group. This study reveals that functionalized SWCNTs could be a promising immersion DNA vaccine carrier in aquaculture.
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immersion vaccination of mandarin fish siniperca chuatsi against infectious spleen and Kidney Necrosis virus with a swcnts based subunit vaccine
Fish & Shellfish Immunology, 2019Co-Authors: Zhao Zhao, Gao Xue Wang, Qiang Lin, Chen Zhang, Zhibin Huang, Yijun Jia, Dekui Qiu, Bin ZhuAbstract:Infectious spleen and Kidney Necrosis virus (ISKNV) cause a high mortality disease which lead to significant economic loss on mandarin fish in China. There is no effective drug or vaccine against this fatal disease at present. Meanwhile, many drugs and vaccines had no effect in many cases account of several impenetrable barriers (cell, skin and gastrointestinal tract). Here we reported an immersion subunit vaccine system (SWCNTs-MCP) encoding MCP gene of ISKNV based on single-walled carbon nanotubes (SWCNTs). To evaluate its efficacy against ISKNV, we found a stronger and longer duration immune response (serum antibody production, enzyme activities and immune-related genes expression) can be induced in fish vaccinated with SWCNTs-MCP in comparison with those vaccinated with MCP alone. Importantly, SWCNTs can increase the immune protective effect of naked subunit vaccine by ca. 23.8%. Thereby, this study demonstrates that SWCNTs as a promising carrier for subunit vaccine might be used to vaccinate large-scale juvenile mandarin fish by bath administration approach.
