鲤春病毒血症(spring virernia of carp,SVC)是由鲤春病毒血症病毒(carp spring virernia virus, CSVV)引起的多种鲤科鱼类高死亡率的病毒病,一般发生在春季,暴发后世界渔业经济损失严重。SVC是世界动物卫生组织(Office International D...鲤春病毒血症(spring virernia of carp,SVC)是由鲤春病毒血症病毒(carp spring virernia virus, CSVV)引起的多种鲤科鱼类高死亡率的病毒病,一般发生在春季,暴发后世界渔业经济损失严重。SVC是世界动物卫生组织(Office International Des Epizooties,OIE)必报的重要疫病,也是我国2008年最新颁布的动物疫病病种名录中的一类动物疫病。本文阐述了CSVV的病原生物学特性、流行病学、病理学,介绍国内外CSVV的检测技术手段、防治方法及疫苗的研发等,为CSVV的防治提供参考。展开更多
糖蛋白(Glycoprotein,G)作为鲤春病毒血症病毒(Spring Virernia of Carp Virus,SVCV)主要的抗原蛋白,已成为现阶段SVCV病毒检测、抗体制备以及疫苗研制的热点。为了对其进行酵母表面展示,研究以SVCVshlj1分离株基因组为模板,通过RT-PCR...糖蛋白(Glycoprotein,G)作为鲤春病毒血症病毒(Spring Virernia of Carp Virus,SVCV)主要的抗原蛋白,已成为现阶段SVCV病毒检测、抗体制备以及疫苗研制的热点。为了对其进行酵母表面展示,研究以SVCVshlj1分离株基因组为模板,通过RT-PCR技术,体外扩增获得SVCV表面糖蛋白的基因开放阅读框(1530 bp)片段,将其克隆至酵母表面展示载体pYD1,构建重组质粒pYD1-G。利用电转化方法将重组质粒pYD1-G导入酿酒酵母EBY100感受态细胞,经YNB选择培养基筛选和菌液PCR的鉴定,挑选出阳性转化子(命名为EBY100-pYD1-G),对其进行2%半乳糖诱导。利用细胞免疫荧光和流式细胞仪检测G蛋白的酵母表面展示情况。细胞免疫荧光结果显示,诱导后的酵母细胞EBY100-pYD1-G能产生特异性红色荧光,且随着诱导时间的增加,红色荧光的酵母细胞所占比例不断增加,各组之间差异显著(P<0.05)。流式细胞仪检测结果显示,酵母细胞的荧光强度与诱导时间呈正比,其中诱导48h与72h的酵母细胞荧光强度不存在显著差异,基本趋于稳定不变的状态。因此,选取诱导48h为酵母表面展示的最佳诱导时间。上述研究结果表明SVCV的G蛋白已经成功展示于酿酒酵母细胞表面,研究为鲤春病毒血症酵母口服疫苗的研发奠定了前期基础。展开更多
[ Objective] The aim was to explore the effects of compound Chinese medicine on growth and antioxidant capacity of Acipenser schrenckii, and to provide a foundation for the application of traditional Chinese medicine ...[ Objective] The aim was to explore the effects of compound Chinese medicine on growth and antioxidant capacity of Acipenser schrenckii, and to provide a foundation for the application of traditional Chinese medicine as immune additives in sturgeon aquaculture production. [Methods] Three kinds of compound Chinese medicines （respectively called A, B and C） were added into basic material feed as feed additives, continuous fed 1 ^＋ age Acipenser schrenckii for 35 d, then weight gain rate was determined and superoxide dismutase （SOD） activity, malondialdehyde （MDA） content, total antioxidant capacity （T-AOC） and glutathione peroxidase （GSH-PX） activity in serum and liver were detected. [ Results ] Chinese medicine formulae could increase the superoxide dismutase （SOD） activity, total antioxidant capacity （ToAOC） and glutathione per- oxidase （GSH-PX） in serum and liver of sturgeon. Especially SOD enzyme activity of serum in the three formulae groups was significantly higher （ P 〈 0.05） than control group, SOD enzyme activities of fish liver in group A and C were significantly higher （ P 〈 0.05） than control group; T-AOC activities of serum and liver in the three formulae groups were both incressed, but had no significant differences with that of the control group; only GSH-PX activities of fish serum in group B and C had significant differences （P〈0.05） with that of control group, GSH-PX activities of liver in the three formulae groups had no significant differences with that of control group; and traditional Chinese medicine could significantly reduce malondialdehyde （MDA） content of sturgeon serum, MDA contents of serum and liver in three formulae groups had significant differences （P 〈 0.05） with that of control group. [ Conclusion] The selected three kinds of compound Chinese medicine additives had different degree of ability to improve sturgeon antioxidant function.展开更多
A total of 59 isolates of Aeromonas hydrophila were collected from common carp suffering from freshwater fish hemorrhage disease in 13 fishing grounds in the northeast China, and their phenotypic and genetic character...A total of 59 isolates of Aeromonas hydrophila were collected from common carp suffering from freshwater fish hemorrhage disease in 13 fishing grounds in the northeast China, and their phenotypic and genetic characteristics were investigated. All of the isolates were identified as A. hydrophila by traditional biochemical method and yielded a 686-bp DNA fragment of the 16S rDNA gene in the PCR experiments. Collected strains were also evaluated for their susceptibility to 17 different antibiotics. The isolates showed an even trend of the resistance and sensitivity to drugs, highly sensitive to antibiotics, such as Levofloxacin, PolymyxinB, Ofloxacin and resistant to antibiotics, such as Bristopen, Lincomycin, Ampicillin, Teicoplanin. Evaluation of genetic diversity was performed on all isolates by molecular typing with enterobacterial repetitive intergenic consensus （ERIC-PCR） method. The results showed that three different types, i.e. type I, ]I, and type ~I, were found in 59 isolates and type III accounted for a large proportion of 54.84%. There was no dominant difference between the tendency of the isolates of Heilongjiang Province and Jilin Province in these three types, which showed DI 〉 I 〉 I, while the isolates of Liaoning Province showed III 〉 II 〉 I. The percentage of different types in different provinces varied in each other; however, they didn＇t show any obvious regional or cluster-specific branches. In conclusion, the ability to distinguish Aeromonas hydrophila strains from diseased common carp with ERIC-PCR would be useful for epidemiological investigation and population genetic analysis of this pathogen in China.展开更多
The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four...The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.展开更多
文摘鲤春病毒血症(spring virernia of carp,SVC)是由鲤春病毒血症病毒(carp spring virernia virus, CSVV)引起的多种鲤科鱼类高死亡率的病毒病,一般发生在春季,暴发后世界渔业经济损失严重。SVC是世界动物卫生组织(Office International Des Epizooties,OIE)必报的重要疫病,也是我国2008年最新颁布的动物疫病病种名录中的一类动物疫病。本文阐述了CSVV的病原生物学特性、流行病学、病理学,介绍国内外CSVV的检测技术手段、防治方法及疫苗的研发等,为CSVV的防治提供参考。
基金Public welfare industry(agriculture)research special funds(201003055)Central public research institutes basic scientific research business expenses special(201003)
文摘[ Objective] The aim was to explore the effects of compound Chinese medicine on growth and antioxidant capacity of Acipenser schrenckii, and to provide a foundation for the application of traditional Chinese medicine as immune additives in sturgeon aquaculture production. [Methods] Three kinds of compound Chinese medicines （respectively called A, B and C） were added into basic material feed as feed additives, continuous fed 1 ^＋ age Acipenser schrenckii for 35 d, then weight gain rate was determined and superoxide dismutase （SOD） activity, malondialdehyde （MDA） content, total antioxidant capacity （T-AOC） and glutathione peroxidase （GSH-PX） activity in serum and liver were detected. [ Results ] Chinese medicine formulae could increase the superoxide dismutase （SOD） activity, total antioxidant capacity （ToAOC） and glutathione per- oxidase （GSH-PX） in serum and liver of sturgeon. Especially SOD enzyme activity of serum in the three formulae groups was significantly higher （ P 〈 0.05） than control group, SOD enzyme activities of fish liver in group A and C were significantly higher （ P 〈 0.05） than control group; T-AOC activities of serum and liver in the three formulae groups were both incressed, but had no significant differences with that of the control group; only GSH-PX activities of fish serum in group B and C had significant differences （P〈0.05） with that of control group, GSH-PX activities of liver in the three formulae groups had no significant differences with that of control group; and traditional Chinese medicine could significantly reduce malondialdehyde （MDA） content of sturgeon serum, MDA contents of serum and liver in three formulae groups had significant differences （P 〈 0.05） with that of control group. [ Conclusion] The selected three kinds of compound Chinese medicine additives had different degree of ability to improve sturgeon antioxidant function.
基金Supported by the Central-Level Non-profit Scientific Research Institutes Special Funds (201104)the Natural Science Foundation of Heilongjiang Province of China (C201140)
文摘A total of 59 isolates of Aeromonas hydrophila were collected from common carp suffering from freshwater fish hemorrhage disease in 13 fishing grounds in the northeast China, and their phenotypic and genetic characteristics were investigated. All of the isolates were identified as A. hydrophila by traditional biochemical method and yielded a 686-bp DNA fragment of the 16S rDNA gene in the PCR experiments. Collected strains were also evaluated for their susceptibility to 17 different antibiotics. The isolates showed an even trend of the resistance and sensitivity to drugs, highly sensitive to antibiotics, such as Levofloxacin, PolymyxinB, Ofloxacin and resistant to antibiotics, such as Bristopen, Lincomycin, Ampicillin, Teicoplanin. Evaluation of genetic diversity was performed on all isolates by molecular typing with enterobacterial repetitive intergenic consensus （ERIC-PCR） method. The results showed that three different types, i.e. type I, ]I, and type ~I, were found in 59 isolates and type III accounted for a large proportion of 54.84%. There was no dominant difference between the tendency of the isolates of Heilongjiang Province and Jilin Province in these three types, which showed DI 〉 I 〉 I, while the isolates of Liaoning Province showed III 〉 II 〉 I. The percentage of different types in different provinces varied in each other; however, they didn＇t show any obvious regional or cluster-specific branches. In conclusion, the ability to distinguish Aeromonas hydrophila strains from diseased common carp with ERIC-PCR would be useful for epidemiological investigation and population genetic analysis of this pathogen in China.
基金Supported by the National Natural Science Foundation of China(31802345)China Postdoctoral Science Foundation(2018M630893)Heilongjiang Province Postdoctoral Science Foundation(LBH-Z18275)。
文摘The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.
