一株中等毒力牛种布鲁氏菌的鉴定和毒力测定

doi:10.3864/j.issn.0578-1752.2014.13.017

Abstract

Abstract: 【Objective】The objective of this study is to explore the biological characteristics and assess the virulence of a B. abortus isolate (strain 343).【Method】The culture of B. abortus 343 was diluted and cultivated to single clone on Tryptic Soy Agar (TSA) for morphological observation, and it was then stained with Gram’s and Koch’s methods. Biochemical characteristics tests (including the growth ability test on thionine and alkaline azaleine, the dependence on CO2, and H2S release test) , single phase specific serum (B. abortus specific anti-serum A, B. melitensis specific anti-serum M and Brucella rough-type specific anti-serum R) agglutination tests and AMOS-PCR were used to identify its comprehensive characteristics. Mice and guinea pigs were infected with a certain dose of B. abortus 343 to determine the virulence of the isolate. The survival time of B. abortus in mice, the isolated number per gram of the spleen from the infected guinea pigs, and the minimum infecting dose (MID) for guinea pigs were tested.【Result】B. abortus 343 is a smooth-type, Gram-negative, Koch's staining red strain, which can reflect glaucous radiance from its surface culture. It releases H2S during its metabolism, and grows on TSA containing thionine or alkaline azaleine, and the growth of which was not dependent on CO2. The antigen prepared from B. abortus 343 was positive with A-type serum in plate agglutination test, and it could induce the immunized mice to produce the specific antibody. The bacteria were isolated from the spleen of the tested mice 29 weeks post infection with a dose of 1×105 CFU. While infected with a dose of 1×109 CFU to the female guinea pigs, 2.4×105 CFU - 1.2×106 CFU could be cultured from spleen per gram 14 days post infection. It could induce specific smooth-type antibody from all guinea pigs infected with a dose of 1×105 CFU, and 1 month post infection the titers of serum were high from 320 to 1 280 in agglutination test. To guinea pigs, the MID is about 40 CFU. 【Conclusion】A middle virulence B. abortus strain, which can be used as a reference strain in the further study was identified. As a new isolate, the detailed identified B. abortus 343 strain is a good supplement to the brucella resources.

Key words: Brucella , AMOS-PCR , minimum infecting dose (MID) , virulence

DING Jia-Bo-1, WANG Fang-1, YANG Hong-Jun-2, WANG Nan-1, ZHU Liang-Quan-1, GU Jin-Hua-1, ZHANG Guang-Chuan-1, WANG Hai-Guang-1, ZHAO Peng-3, CHENG Jun-Sheng-1, MAO Kai-Rong-1, FENG Yu-3. Identification and Virulence Assessment for a Strain of Brucella abortus[J].Scientia Agricultura Sinica, 2014, 47(13): 2652-2658.

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Identification and Virulence Assessment for a Strain of Brucella abortus

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