牦牛漫游球菌的分离与生物学特性鉴定
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摘要
为探究1只牦牛屠宰后肝脏有干酪样结节形成的原因,对肝脏进行细菌的分离培养,对分离到的1株细菌的16S rRNA和16S～23S rRNA基因序列用通用引物扩增和并用NCBI blast和MEGA 7进行序列分析,并用Vitek 2全自动微生物鉴定仪进行生化表型的鉴定,用K-B法进行药敏试验,结果显示:该菌的16S rRNA为1 524 bp,NCBI GenBank登录号为MG753544.1,与水獭漫游球菌的同源性较高,16S～23S rRNA有大小不同的2个片段,分别为522,425 bp,NCBI GenBank登录号分别为MG758122.1和MG758123.1。Vitek 2全自动微生物鉴定仪鉴定为迟缓葡萄球菌。该菌对妥布霉素、头孢哌酮、米诺环素等多种药物高度敏感,对苯唑西林、头孢西丁有抗性。结果表明,分离出的细菌为漫游球菌,该菌是否具有致病性还有待进一步试验验证。
To investigate the formation cause of caseous nodules in the liver of a yak after slaughtering.The bacteria from the pathological liver were isolated and cultured.The 16 S rRNA and 16 S-23 S rRNA gene sequences of the bacteria were amplified by PCR with universal primers.The amplified sequence was analyzed with NCBI blast and MEGA 7.The biochemical phenotypes were identified by Vitek 2 Compact and drug susceptibility test was performed by K-B method.The amplification lengths of 16 S rRNA is 1 524 bp and sequence accession number is MG753544.1.The sequence similarity of 16 S rRNA of clinical isolation to Vagococcus lutrae was 99%.The 16 S-23 S rRNA has two fragments with different sizes,522 bp and 425 bp respectively.The accession number is MG758122.1 and MG758123.1.It was been identified as Staphylococcus lentus by Vitek 2 Compact,with a confidence level of 96%.The bacteria is sensitive to tobramycin,cefoperazone,minocycline and resistant to oxacillin,cefoxitin.The isolated bacteria is Vagococcus sp.Whether the bacteria is involved in the formation of liver lesions remains to be verified by further tests.
To investigate the formation cause of caseous nodules in the liver of a yak after slaughtering.The bacteria from the pathological liver were isolated and cultured.The 16 S rRNA and 16 S-23 S rRNA gene sequences of the bacteria were amplified by PCR with universal primers.The amplified sequence was analyzed with NCBI blast and MEGA 7.The biochemical phenotypes were identified by Vitek 2 Compact and drug susceptibility test was performed by K-B method.The amplification lengths of 16 S rRNA is 1 524 bp and sequence accession number is MG753544.1.The sequence similarity of 16 S rRNA of clinical isolation to Vagococcus lutrae was 99%.The 16 S-23 S rRNA has two fragments with different sizes,522 bp and 425 bp respectively.The accession number is MG758122.1 and MG758123.1.It was been identified as Staphylococcus lentus by Vitek 2 Compact,with a confidence level of 96%.The bacteria is sensitive to tobramycin,cefoperazone,minocycline and resistant to oxacillin,cefoxitin.The isolated bacteria is Vagococcus sp.Whether the bacteria is involved in the formation of liver lesions remains to be verified by further tests.
引文
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[2] CHANG H,DAI F,DUAN B,et al.Isolation and characterization of Vagococcus carniphilus from diseased crucian carp[J].Biotechnol Biotechnol Equip,2017(1):1-6.
[3] TEIXEIRA L M,CARVALHO M G,MERQUIOR V L,et al.Phenotypic and genotypic characterization of Vagococcus fluvialis,including strains isolated from human sources[J].J Clin Microbiol,1997,35(11):2778-2781.
[4] 杨苗,陈世界,林华,等.一株河流漫游球菌的分离鉴定[J].四川畜牧兽医,2015(9):30-32.
[5] CHANDEL K,PARIKH R Y,MENDKI M J,et al.Isolation and characterization of Vagococcus sp from midgut of Culex quinquefasciatus (Say) mosquito[J].J Vector Borne Dis,2015,52(1):52.
[6] WANG L,CUI Y S,KWON C S,et al.Vagococcus acidifermentans sp.nov.,isolated from an acidogenic fermentation bioreactor[J].Int J Syst Evol Microbiol,2011,61(5):1123-1126.
[7] LEBRETON F,VALENTINO M D,DUNCAN L B,et al.High-quality draft genome sequence of Vagococcus lutrae strain LBD1,isolated from the largemouth bass micropterus salmoides[J].Genome Announc,2013,1(6):e01087-13.
[8] GAO J,BAO H Y,XIN M X,et al.Characterization of a bioflocculant from a newly isolated Vagococcus sp.W31[J].J Zhejiang Univ Sci B,2006,7(3):186-192.
[9] CHANG H C,WEI Y F,DIJKSHOORN L,et al.Species-level identification of isolates of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex by sequence analysis of the 16S-23S rRNA gene spacer region[J].J Clin Microbiol,2005(43):1632-1639.
[10] 李仲兴.革兰阳性球菌与临床感染[M].北京:科学出版社,2007:509-514.
[11] WALLBANKS S,MARTINEZMURCIA A J,FRYER J L,et al.16S rRNA sequence determination for members of the genus Carnobacterium and related lactic acid bacteria and description of Vagococcus salmoninarum sp.nov[J].Int J Syst Bacteriol,1990,40(3):224.
[12] 房海,陈翠珍,张晓君,水产养殖动物病原细菌学[M].北京:中国农业出版社,2010:640-642.
[13] GURTLER V,STANISICH V A.New approaches to typing and identification of bacteria using the 16S-23S rDNA spacer region[J].Microbiology,1996(142):3-16.