Coagulation test, in principle, is an immunodiagnostics technique, in which immunoglobuline G of antibody is bound to protein A from Staphylococcus aureus. The aim of study is to develop a rapid test kit for detecting...Coagulation test, in principle, is an immunodiagnostics technique, in which immunoglobuline G of antibody is bound to protein A from Staphylococcus aureus. The aim of study is to develop a rapid test kit for detecting iridovirus infection in fish. Method was summarized as follows： （1） vaccine of iridovirus was injected to rabbit four times with a dosage as 0.5 mL, 1 mL, 2mL, 3 mL each week. Serum was collected at the fifth week as a coagglutination test kit; （2） through the positif polymerase chain reaction （PCR） test, the kidney and spleen sample infected with iridovirus are homogenized by using the phosphate buffered saline （PBS） solution of pH 7.2 with ratio 1：2 （WN）; （3） the supernatant material is collected after centrifugation at 8,000 rpm for 15 min; （4） filtrate/supernatant from sample was dropped on a slide an added with coagglutination test kit with the same volume （l：l）; （5） the agglutination observation is done after the 30, 60 and 90 min incubate at room temperature. The coagglutination test gave positive result in 25% of the test samples.展开更多
A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus (LYCIV) is described, which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated fr...A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus (LYCIV) is described, which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen. Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus (RSIV) and sea bass iridovirus (SBIV), suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone. The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes, the expected fragment was detected from spleen DNA samples of infected fishes, whereas no fragments were amplified from healthy fish spleen DNA, white spot syndrome baculoviruses (WSBV) DNA and pseudorabies virus (PRV) DNA. Detection limit of this method was 10(-7) ng positive plasmid DNA containing target sequence, equal to about 100 virions. In the infected experiment, first positive detection (1/4) appeared at Day 3 post-infection, all fish (4/4) tested positive at Day 7, however obvious symptoms were observed at Day 8, so LYCIV infection could be detected prior to the appearance of obvious symptoms. These results indicate that this PCR method could be used for early, rapid and specific detection of LYCIV infection.展开更多
Iridovirus poses a substantial threat to global aquaculture due to its high mortality rate;however,the molecular mechanisms underpinning its pathogenesis are not well elucidated.Here,a multi-omics approach was applied...Iridovirus poses a substantial threat to global aquaculture due to its high mortality rate;however,the molecular mechanisms underpinning its pathogenesis are not well elucidated.Here,a multi-omics approach was applied to groupers infected with Singapore grouper iridovirus(SGIV),focusing on the roles of key metabolites.Results showed that SGIV induced obvious histopathological damage and changes in metabolic enzymes within the liver.Furthermore,SGIV significantly reduced the contents of lipid droplets,triglycerides,cholesterol,and lipoproteins.Metabolomic analysis indicated that the altered metabolites were enriched in 19 pathways,with a notable down-regulation of lipid metabolites such as glycerophosphates and alpha-linolenic acid(ALA),consistent with disturbed lipid homeostasis in the liver.Integration of transcriptomic and metabolomic data revealed that the top enriched pathways were related to cell growth and death and nucleotide,carbohydrate,amino acid,and lipid metabolism,supporting the conclusion that SGIV infection induced liver metabolic reprogramming.Further integrative transcriptomic and proteomic analysis indicated that SGIV infection activated crucial molecular events in a phagosome-immune depression-metabolism dysregulation-necrosis signaling cascade.Of note,integrative multi-omics analysis demonstrated the consumption of ALA and linoleic acid(LA)metabolites,and the accumulation of L-glutamic acid(GA),accompanied by alterations in immune,inflammation,and cell death-related genes.Further experimental data showed that ALA,but not GA,suppressed SGIV replication by activating antioxidant and anti-inflammatory responses in the host.Collectively,these findings provide a comprehensive resource for understanding host response dynamics during fish iridovirus infection and highlight the antiviral potential of ALA in the prevention and treatment of iridoviral diseases.展开更多
Turbot(Scophthalmus maximus L.) reddish body iridovirus(TRBIV) was propagated in turbot fin cells(TF cells) and inactivated as the TRBIV vaccine with its protection efficiency evaluated in this study.TF cells were cul...Turbot(Scophthalmus maximus L.) reddish body iridovirus(TRBIV) was propagated in turbot fin cells(TF cells) and inactivated as the TRBIV vaccine with its protection efficiency evaluated in this study.TF cells were cultured in 10% bovine calf serum(BCS)-containing MEM medium(pH7.0) at 22℃,in which TRBIV propagated to a titer as high as 105.6 TCID50 mL-1.The TRBIV was inactivated with 0.1% formalin and formulated with 0.5% aluminum hydroxide.The inactivated vaccine caused neither cytopathogenic effect(CPE) on TF cells nor pathogenic effect on turbots.After being administered with the vaccine twice via muscle injection,the turbot developed high-tittered TRBIV neutralizing antibodies in a dose-dependent manner.The vaccine protected the turbot from dying with an immunoprotection rate of 83.3% as was determined via subcutaneous vaccination in the laboratory and 90.5% via bath vaccination in turbot farms,respectively.The inactivated vaccine was very immunogenic,efficiently preventing tur-bot from death.It holds the potential of being applied in aquaculture.展开更多
Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of t...Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of the cause of an outbreak of illness caused by iridovirus in the field, so that remedial action can be taken quickly and appropriately to minimize the impact of wider losses. Samples were taken from the grouper and pomfret star farms that are experiencing outbreaks of infectious diseases in the months from May to August 2015, Tanjungpinang, Indonesia. The sick and allegedly attacked by iridovirus samples showed abnormal swimming clinical symptoms, weakness and the swollen spleen. The swollen spleen of sick fish created suspension in phosphate buffered saline （PBS） with pH 7.2, and then centrifuged at 8,000 rpm for I0 rain. The supernatant after centrifuge was used as the test sample. On a clean object glass, 50 μL of the supematant was treated with 50 μL kit co-agglutination pre-prepared. The positive results were shown by the agglutination reaction after 10-15 rain, while as a negative control, PBS was reacted with co-agglutination kit that looked homogeneous （no agglutination）. It was showed that the grouper （Epinepkelus sp.） on four farms and pomfret star （Thracinotus blochii） on one farm that experienced an outbreak of infectious disease in Tanjungpinang showed positively infected iridovirus. The same positive iridovirus result was also demonstrated by examination using polymerase chain reaction （PCR） at 570 bp. So, the causative agent of plague on grouper and pomfret star was iridovirus. In addition, the co-agglutination test based on serology is more quick, cheap and accurate.展开更多
There are a variety of viral pathogens that cause disease in mosquitoes with most belonging to three major groups. The most common viruses of mosquitoes are the baculoviruses (DBVs) (Baculoviridae: Deltabaculovirus), ...There are a variety of viral pathogens that cause disease in mosquitoes with most belonging to three major groups. The most common viruses of mosquitoes are the baculoviruses (DBVs) (Baculoviridae: Deltabaculovirus), cytoplasmic polyhedrosis viruses (CPVs) (Reoviridae: Cypovirus) and the iridoviruses (MIVs) (Iridoviridae: Chloriridovirus). Baculoviruses and iridoviruses are DNA viruses while cypoviruses are the main RNA viruses in mosquitoes. This review presents an overview of the current status and recent advancements in understanding the biology and molecular features of mosquito pathogenic viruses.展开更多
大菱鲆病毒性红体病(Viral reddish body syndrome, VRBS)是一种新发现的感染我国养殖大菱鲆的流行性疾病.本文描述了病鱼的外观症状和解剖病理特征,报道了该病的病原及疾病流行情况调查结果.外观检查发现,病鱼的体表无明显损伤,但腹面...大菱鲆病毒性红体病(Viral reddish body syndrome, VRBS)是一种新发现的感染我国养殖大菱鲆的流行性疾病.本文描述了病鱼的外观症状和解剖病理特征,报道了该病的病原及疾病流行情况调查结果.外观检查发现,病鱼的体表无明显损伤,但腹面沿脊椎骨附近皮下淤血、发红,鳍及鳍基部充血、发红;病鱼贫血,血液凝固性差;肾脏肿大,呈灰白色.组织病理学研究显示病鱼脾组织中存在大量肥大细胞,电镜切片中可见大量平均直径约125 nm的二十面体病毒粒子,即大菱鲆红体病虹彩病毒(Turbot reddish body iridovirus, TRBIV).将过滤除菌的含TRBIV的病鱼脾组织匀浆液,经腹腔注射进行人工感染,感染鱼在3周内的累积死亡率达85.7%,死亡大菱鲆表现出腹面和鳍边发红等外观症状,并在感染鱼脾组织切片中观察到大量同样的病毒粒子,由此证实TRBIV是我国养殖大菱鲆病毒性红体病的病原.疾病流行情况调查显示,该病多在养成期大菱鲆中流行,高发季节为每年的8～12月.展开更多
文摘Coagulation test, in principle, is an immunodiagnostics technique, in which immunoglobuline G of antibody is bound to protein A from Staphylococcus aureus. The aim of study is to develop a rapid test kit for detecting iridovirus infection in fish. Method was summarized as follows： （1） vaccine of iridovirus was injected to rabbit four times with a dosage as 0.5 mL, 1 mL, 2mL, 3 mL each week. Serum was collected at the fifth week as a coagglutination test kit; （2） through the positif polymerase chain reaction （PCR） test, the kidney and spleen sample infected with iridovirus are homogenized by using the phosphate buffered saline （PBS） solution of pH 7.2 with ratio 1：2 （WN）; （3） the supernatant material is collected after centrifugation at 8,000 rpm for 15 min; （4） filtrate/supernatant from sample was dropped on a slide an added with coagglutination test kit with the same volume （l：l）; （5） the agglutination observation is done after the 30, 60 and 90 min incubate at room temperature. The coagglutination test gave positive result in 25% of the test samples.
文摘A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus (LYCIV) is described, which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen. Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus (RSIV) and sea bass iridovirus (SBIV), suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone. The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes, the expected fragment was detected from spleen DNA samples of infected fishes, whereas no fragments were amplified from healthy fish spleen DNA, white spot syndrome baculoviruses (WSBV) DNA and pseudorabies virus (PRV) DNA. Detection limit of this method was 10(-7) ng positive plasmid DNA containing target sequence, equal to about 100 virions. In the infected experiment, first positive detection (1/4) appeared at Day 3 post-infection, all fish (4/4) tested positive at Day 7, however obvious symptoms were observed at Day 8, so LYCIV infection could be detected prior to the appearance of obvious symptoms. These results indicate that this PCR method could be used for early, rapid and specific detection of LYCIV infection.
基金supported by the National Natural Science Foundation of China(31930115,32173007)China Agriculture Research System of MOF and MARA(CARS-47-G16)Basic and Applied Basic Research Foundation of Guangdong Province(2022A1515010595)。
文摘Iridovirus poses a substantial threat to global aquaculture due to its high mortality rate;however,the molecular mechanisms underpinning its pathogenesis are not well elucidated.Here,a multi-omics approach was applied to groupers infected with Singapore grouper iridovirus(SGIV),focusing on the roles of key metabolites.Results showed that SGIV induced obvious histopathological damage and changes in metabolic enzymes within the liver.Furthermore,SGIV significantly reduced the contents of lipid droplets,triglycerides,cholesterol,and lipoproteins.Metabolomic analysis indicated that the altered metabolites were enriched in 19 pathways,with a notable down-regulation of lipid metabolites such as glycerophosphates and alpha-linolenic acid(ALA),consistent with disturbed lipid homeostasis in the liver.Integration of transcriptomic and metabolomic data revealed that the top enriched pathways were related to cell growth and death and nucleotide,carbohydrate,amino acid,and lipid metabolism,supporting the conclusion that SGIV infection induced liver metabolic reprogramming.Further integrative transcriptomic and proteomic analysis indicated that SGIV infection activated crucial molecular events in a phagosome-immune depression-metabolism dysregulation-necrosis signaling cascade.Of note,integrative multi-omics analysis demonstrated the consumption of ALA and linoleic acid(LA)metabolites,and the accumulation of L-glutamic acid(GA),accompanied by alterations in immune,inflammation,and cell death-related genes.Further experimental data showed that ALA,but not GA,suppressed SGIV replication by activating antioxidant and anti-inflammatory responses in the host.Collectively,these findings provide a comprehensive resource for understanding host response dynamics during fish iridovirus infection and highlight the antiviral potential of ALA in the prevention and treatment of iridoviral diseases.
基金supported by the National High Technology Research and Development Program of China(863 Program)(2006AA10A401)
文摘Turbot(Scophthalmus maximus L.) reddish body iridovirus(TRBIV) was propagated in turbot fin cells(TF cells) and inactivated as the TRBIV vaccine with its protection efficiency evaluated in this study.TF cells were cultured in 10% bovine calf serum(BCS)-containing MEM medium(pH7.0) at 22℃,in which TRBIV propagated to a titer as high as 105.6 TCID50 mL-1.The TRBIV was inactivated with 0.1% formalin and formulated with 0.5% aluminum hydroxide.The inactivated vaccine caused neither cytopathogenic effect(CPE) on TF cells nor pathogenic effect on turbots.After being administered with the vaccine twice via muscle injection,the turbot developed high-tittered TRBIV neutralizing antibodies in a dose-dependent manner.The vaccine protected the turbot from dying with an immunoprotection rate of 83.3% as was determined via subcutaneous vaccination in the laboratory and 90.5% via bath vaccination in turbot farms,respectively.The inactivated vaccine was very immunogenic,efficiently preventing tur-bot from death.It holds the potential of being applied in aquaculture.
文摘Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of the cause of an outbreak of illness caused by iridovirus in the field, so that remedial action can be taken quickly and appropriately to minimize the impact of wider losses. Samples were taken from the grouper and pomfret star farms that are experiencing outbreaks of infectious diseases in the months from May to August 2015, Tanjungpinang, Indonesia. The sick and allegedly attacked by iridovirus samples showed abnormal swimming clinical symptoms, weakness and the swollen spleen. The swollen spleen of sick fish created suspension in phosphate buffered saline （PBS） with pH 7.2, and then centrifuged at 8,000 rpm for I0 rain. The supernatant after centrifuge was used as the test sample. On a clean object glass, 50 μL of the supematant was treated with 50 μL kit co-agglutination pre-prepared. The positive results were shown by the agglutination reaction after 10-15 rain, while as a negative control, PBS was reacted with co-agglutination kit that looked homogeneous （no agglutination）. It was showed that the grouper （Epinepkelus sp.） on four farms and pomfret star （Thracinotus blochii） on one farm that experienced an outbreak of infectious disease in Tanjungpinang showed positively infected iridovirus. The same positive iridovirus result was also demonstrated by examination using polymerase chain reaction （PCR） at 570 bp. So, the causative agent of plague on grouper and pomfret star was iridovirus. In addition, the co-agglutination test based on serology is more quick, cheap and accurate.
文摘There are a variety of viral pathogens that cause disease in mosquitoes with most belonging to three major groups. The most common viruses of mosquitoes are the baculoviruses (DBVs) (Baculoviridae: Deltabaculovirus), cytoplasmic polyhedrosis viruses (CPVs) (Reoviridae: Cypovirus) and the iridoviruses (MIVs) (Iridoviridae: Chloriridovirus). Baculoviruses and iridoviruses are DNA viruses while cypoviruses are the main RNA viruses in mosquitoes. This review presents an overview of the current status and recent advancements in understanding the biology and molecular features of mosquito pathogenic viruses.
文摘大菱鲆病毒性红体病(Viral reddish body syndrome, VRBS)是一种新发现的感染我国养殖大菱鲆的流行性疾病.本文描述了病鱼的外观症状和解剖病理特征,报道了该病的病原及疾病流行情况调查结果.外观检查发现,病鱼的体表无明显损伤,但腹面沿脊椎骨附近皮下淤血、发红,鳍及鳍基部充血、发红;病鱼贫血,血液凝固性差;肾脏肿大,呈灰白色.组织病理学研究显示病鱼脾组织中存在大量肥大细胞,电镜切片中可见大量平均直径约125 nm的二十面体病毒粒子,即大菱鲆红体病虹彩病毒(Turbot reddish body iridovirus, TRBIV).将过滤除菌的含TRBIV的病鱼脾组织匀浆液,经腹腔注射进行人工感染,感染鱼在3周内的累积死亡率达85.7%,死亡大菱鲆表现出腹面和鳍边发红等外观症状,并在感染鱼脾组织切片中观察到大量同样的病毒粒子,由此证实TRBIV是我国养殖大菱鲆病毒性红体病的病原.疾病流行情况调查显示,该病多在养成期大菱鲆中流行,高发季节为每年的8～12月.
