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在哥伦比亚鉴定出牛源布鲁氏菌流产亚种生物变种4。

Identification of Brucella abortus biovar 4 of bovine origin in Colombia.

作者信息

Torres Higuera Ligia D, Jiménez Velásquez Sabrina Del C, Rodríguez Bautista José L, Patiño Burbano Rocio E

机构信息

Corporación Colombiana de Investigación Agropecuaria AGROSAVIA, Centro de investigación Tibaitatá, Km 14 vía Mosquera-Bogotá, Mosquera, Colombia.

Programa de Pós-graduação em Ciências Veterinárias, Federal Rural University of Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil.

出版信息

Rev Argent Microbiol. 2019 Jul-Sep;51(3):221-228. doi: 10.1016/j.ram.2018.08.002. Epub 2018 Dec 12.

Abstract

The objective of this study was to identify twelve Brucella abortus isolates of bovine origin from the department of Nariño in Colombia up to the biovar level. These isolates are included in the collection of the Germplasm Bank of Microorganisms of Animal Health Interest - Bacteria and Virus (BGSA-BV). The identification was carried out through conventional methods such as macro and microscopic morphological descriptions, enzymatic activity, biochemical profile, substrate use and sensitivity to dyes. Complementary genotypic characterization was carried out using multiplex PCR for B. abortus, Brucella melitensis, Brucella ovis, and Brucella suis-Erytritol (AMOS-ERY-PCR), RFLP-IS711, by southern blot hybridization, as well as by the multiple locus variable number of tandem repeat analysis (MLVA) using the ery gene and the insertion sequence IS711 and variable number of tandem repeats (VNTR) as molecular markers. The results of the phenotypic and molecular characterization allowed to identify twelve isolates as B. abortus biovar 4 as well as to differentiate field from vaccine strains. This is the first study on the phenotypic and molecular identification of B. abortus isolates in Colombia. It was concluded that the phenotypic and molecular identification of twelve isolates as B. abortus biovar 4 could be achieved using conventional and molecular techniques with enough resolution power. The identification of these isolates to the biovar level in taxonomic and epidemiological terms will allow the use of this genetic resource as reference strains in future research. This finding constitutes the basis for identifying biotypes not previously reported in the country that might be useful to support brucellosis survey programs in Colombia.

摘要

本研究的目的是将来自哥伦比亚纳里尼奥省的12株牛源流产布鲁氏菌分离株鉴定到生物变种水平。这些分离株保存在动物健康相关微生物种质库——细菌和病毒(BGSA-BV)中。鉴定通过常规方法进行,如宏观和微观形态描述、酶活性、生化特征、底物利用及对染料的敏感性。采用多重PCR对流产布鲁氏菌、马尔他布鲁氏菌、绵羊布鲁氏菌和猪布鲁氏菌-赤藓醇(AMOS-ERY-PCR)、RFLP-IS711进行Southern印迹杂交,并以ery基因、插入序列IS711和可变数目串联重复序列(VNTR)作为分子标记进行多位点可变数目串联重复分析(MLVA),进行补充基因型特征分析。表型和分子特征分析结果表明,12株分离株均为流产布鲁氏菌生物变种4,并能区分田间菌株和疫苗菌株。这是哥伦比亚首次对流产布鲁氏菌分离株进行表型和分子鉴定的研究。研究得出结论,使用具有足够分辨力的常规和分子技术可以实现将12株分离株鉴定为流产布鲁氏菌生物变种4。从分类学和流行病学角度将这些分离株鉴定到生物变种水平,将有助于在未来研究中把这种遗传资源用作参考菌株。这一发现为鉴定该国以前未报告的生物型奠定了基础,这些生物型可能有助于支持哥伦比亚的布鲁氏菌病调查项目。

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