目的对大鲵不明原因死亡进行病原学鉴定,并根据药物敏感实验结果选用抗生素治疗。方法从患病死亡大鲵足部溃烂病变部位及心脏取血采样,分别用液体培养基、普通琼脂平板、血平板进行增菌和画线培养分离,经法国BiomeriruxVITEK R 2全自动...目的对大鲵不明原因死亡进行病原学鉴定,并根据药物敏感实验结果选用抗生素治疗。方法从患病死亡大鲵足部溃烂病变部位及心脏取血采样,分别用液体培养基、普通琼脂平板、血平板进行增菌和画线培养分离,经法国BiomeriruxVITEK R 2全自动微生物分析系统检测。结果与结论引起感染是迟钝爱德华菌(E.tarda),该病原体对庆大霉素、头孢曲松钠和环丙沙星等抗菌药物敏感,用敏感的庆大霉素和头孢曲松钠处理,控制了感染的流行。展开更多
[Objective] The study aimed to establish an approach to high titer Et immune serum preparation by low-aged Japanese rabbits. [Method] Antigen of Et was prepared at first, Japanese rabbits were taken as immunized anima...[Objective] The study aimed to establish an approach to high titer Et immune serum preparation by low-aged Japanese rabbits. [Method] Antigen of Et was prepared at first, Japanese rabbits were taken as immunized animal, and divided into two groups for experiments, one is two-month-old group （T group）, and another is six-month-old group （S group）. Japanese rabbits were continuous by immunized with low-dose by using auricular intravenous method, then immune sera were collected. Immune serum antibody titer was determined with micro-agglutination reaction method. E Resultl Agglutination reaction showed that the Et serum titer of S group is higher than that of T group in the first testing with the same dose. But in the second testing, the serum titer of the T group and and the S group was consistent, [Conclusien] The animals in the T group （Japanese rabbits） were fewer months old, and produced high titer antiserum was which consistent with the S group, which indicated that the method of preparing of high titer Et immune serum by low-aged Japanese rabbits was feasible.展开更多
[Objective] The aim of the research was to investigate the function of large plasmid pEIB202 in the pathogenesis of Edwardsiella tarda EIB202 and to eliminate the plasmid pEIB202,so as to lay the foundation for develo...[Objective] The aim of the research was to investigate the function of large plasmid pEIB202 in the pathogenesis of Edwardsiella tarda EIB202 and to eliminate the plasmid pEIB202,so as to lay the foundation for developing safe and live attenu- ated vaccine against E, tarda. [Method] sacB was used as reverse screening marker to eliminate the plasmid by using homologous recombination technique. [Result] The plasmid pEIB202 was sequenced and it was found that the plasmid encoded multiple resistant genes and some components in type IV secretion system（T4SS）,which sug- gested that the plasmid might be related with the multiple drug-resistance and pathogenicity of E. tarda. The plasmid-eliminated strain EIB202Ap lost the resistance to chloramphenicol and tetracycline,but its growth,virulence and secretion of extra- cellular proteins had no significant difference with wild-type plants. [Conclusion] pEIB202 plasmid is the main reason that caused the multi-drug resistance of EIB202 and might have indirect effects in the pathogenesis of EIB202.展开更多
Edwardsiella tarda has become one of the most important emerging pathogens in aquaculture industry. Therefore, a rapid, reproducible, and sensitive method for detection and quantification of this pathogen is needed ur...Edwardsiella tarda has become one of the most important emerging pathogens in aquaculture industry. Therefore, a rapid, reproducible, and sensitive method for detection and quantification of this pathogen is needed urgently. To achieve this purpose, we developed a TaqMan-based real-time PCR assay for detection and quantification orE. tarda. The assay targets the hemolysin activator HlyB domain protein of E. tarda. Our optimized TaqMan assay is capable of detecting as little as 40 fg of genomic DNA per reaction. A standard curve was generated from the threshold cycle values （y） against log10 （E. tarda genomic DNA concentration） as x. The intra- and inter-assay coefficient of variation （CV） values were less than 2.06% and 1.05% respectively, indicating that the assay had good reproducibility. This method is highly specific to E. tarda strains, as it shows no cross-reactivity to Edwardsiella ictaluri, a member of the same genus, or to nine other fish-pathogenic bacteria species belonging to three other genera. This sensitive and specific real-time PCR assay provides a valuable tool for diagnostic quantitation of E. tarda in clinical samples.展开更多
文摘目的对大鲵不明原因死亡进行病原学鉴定,并根据药物敏感实验结果选用抗生素治疗。方法从患病死亡大鲵足部溃烂病变部位及心脏取血采样,分别用液体培养基、普通琼脂平板、血平板进行增菌和画线培养分离,经法国BiomeriruxVITEK R 2全自动微生物分析系统检测。结果与结论引起感染是迟钝爱德华菌(E.tarda),该病原体对庆大霉素、头孢曲松钠和环丙沙星等抗菌药物敏感,用敏感的庆大霉素和头孢曲松钠处理,控制了感染的流行。
基金Supported by Science and Technology Key Project of Science and Technology Bureau of Tangshan:Study of Rapid Detection and Diag-nosis Technology on Key Aquatic Animal Pathogen(06125401A-3)~~
文摘[Objective] The study aimed to establish an approach to high titer Et immune serum preparation by low-aged Japanese rabbits. [Method] Antigen of Et was prepared at first, Japanese rabbits were taken as immunized animal, and divided into two groups for experiments, one is two-month-old group （T group）, and another is six-month-old group （S group）. Japanese rabbits were continuous by immunized with low-dose by using auricular intravenous method, then immune sera were collected. Immune serum antibody titer was determined with micro-agglutination reaction method. E Resultl Agglutination reaction showed that the Et serum titer of S group is higher than that of T group in the first testing with the same dose. But in the second testing, the serum titer of the T group and and the S group was consistent, [Conclusien] The animals in the T group （Japanese rabbits） were fewer months old, and produced high titer antiserum was which consistent with the S group, which indicated that the method of preparing of high titer Et immune serum by low-aged Japanese rabbits was feasible.
基金Supported by National Technology System for Flatfish Culture Industry(CARS-50)National High Technology Research and Development Program of China(863Program)(2008AA092501)~~
文摘[Objective] The aim of the research was to investigate the function of large plasmid pEIB202 in the pathogenesis of Edwardsiella tarda EIB202 and to eliminate the plasmid pEIB202,so as to lay the foundation for developing safe and live attenu- ated vaccine against E, tarda. [Method] sacB was used as reverse screening marker to eliminate the plasmid by using homologous recombination technique. [Result] The plasmid pEIB202 was sequenced and it was found that the plasmid encoded multiple resistant genes and some components in type IV secretion system（T4SS）,which sug- gested that the plasmid might be related with the multiple drug-resistance and pathogenicity of E. tarda. The plasmid-eliminated strain EIB202Ap lost the resistance to chloramphenicol and tetracycline,but its growth,virulence and secretion of extra- cellular proteins had no significant difference with wild-type plants. [Conclusion] pEIB202 plasmid is the main reason that caused the multi-drug resistance of EIB202 and might have indirect effects in the pathogenesis of EIB202.
基金Supported by the Special Fund for Agro-scientific Research in the Public Interest(No.201103034)the Construction Special Fund of Modern Agriculture and Industrial Technology Research System(No.CARS-47)
文摘Edwardsiella tarda has become one of the most important emerging pathogens in aquaculture industry. Therefore, a rapid, reproducible, and sensitive method for detection and quantification of this pathogen is needed urgently. To achieve this purpose, we developed a TaqMan-based real-time PCR assay for detection and quantification orE. tarda. The assay targets the hemolysin activator HlyB domain protein of E. tarda. Our optimized TaqMan assay is capable of detecting as little as 40 fg of genomic DNA per reaction. A standard curve was generated from the threshold cycle values （y） against log10 （E. tarda genomic DNA concentration） as x. The intra- and inter-assay coefficient of variation （CV） values were less than 2.06% and 1.05% respectively, indicating that the assay had good reproducibility. This method is highly specific to E. tarda strains, as it shows no cross-reactivity to Edwardsiella ictaluri, a member of the same genus, or to nine other fish-pathogenic bacteria species belonging to three other genera. This sensitive and specific real-time PCR assay provides a valuable tool for diagnostic quantitation of E. tarda in clinical samples.
