流产奶牛胎儿肺炎克雷伯氏菌的分离与鉴定
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摘要
从流产奶牛胎儿组织中分离到1株革兰氏阴性短杆菌,采用细菌分离纯化、生化试验、16S rRNA和16S～23S rRNA测序、药敏试验和小鼠致死性试验对其进行了鉴定。结果显示,该分离菌与GenBank中肺炎克雷伯氏菌16S rRNA和16S～23S rRNA基因序列一致性分别高达99%和96%,16S～23S rRNA基因被扩增出3条不同长度条带;该菌生化特性均符合肺炎克雷伯氏菌的生化反应特性;对头孢曲松、阿米卡星、多粘菌素等敏感,对多西环素、四环素、卡拉霉素等中度敏感,对青霉素、红霉素、米诺环素等耐药;对小鼠有很强的致病性。以上结果表明,该分离细菌为肺炎克雷伯氏菌。
A gram-negative bacillus was isolated from tissues of an aborted dairy fetus and identified as Klebsiella pneumoniae by its morphology and culture characteristics, biochemistry test, 16 S rRNA and 16 S-23 S rRNA gene sequencing, drug susceptibility and pathogenicity. The 16 S rRNA gene and 16 S-23 S gene sequences of this isolate shared 99% and 96% homology with Klebsiella pneumoniae gene sequences published in GenBank and its 16 S-23 S gene was amplified as three different length products. The biochemistry test results of this isolate were consistent with Klebsiella pneumoniae. This isolate was virulent to mice. The drug test demonstrated that this isolate was highly susceptible to ceftriaxone, amikacin and polymyxin, susceptible to doxycycline, tetracycline and kalafungin but resistant to penicillin, erythromycin and inocycline. The results indicated that Klebsiella pneumoniae was isolated.
A gram-negative bacillus was isolated from tissues of an aborted dairy fetus and identified as Klebsiella pneumoniae by its morphology and culture characteristics, biochemistry test, 16 S rRNA and 16 S-23 S rRNA gene sequencing, drug susceptibility and pathogenicity. The 16 S rRNA gene and 16 S-23 S gene sequences of this isolate shared 99% and 96% homology with Klebsiella pneumoniae gene sequences published in GenBank and its 16 S-23 S gene was amplified as three different length products. The biochemistry test results of this isolate were consistent with Klebsiella pneumoniae. This isolate was virulent to mice. The drug test demonstrated that this isolate was highly susceptible to ceftriaxone, amikacin and polymyxin, susceptible to doxycycline, tetracycline and kalafungin but resistant to penicillin, erythromycin and inocycline. The results indicated that Klebsiella pneumoniae was isolated.
引文
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[16]周玉龙,李国军,李阳,等.奶牛肺炎克雷伯氏菌的分离与鉴定[J].中国奶牛,2007(6):36-38.
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[2]Hiroi M,Yamazaki F,Harada T,et al.Prevalence of extended-spectrumβ-lactamase-producing Escherichia coli and Klebsiella pneumoniae in food-producing animals[J].J Vet Med Sci,2012,74(2):189-195.
[3]Jian-Li W,Yuan-Yuan S,Shou-Yu G,et al.Serotype and virulence genes of Klebsiella pneumoniae isolated from mink and its pathogenesis in mice and mink[J].Sci Rep,2017,7(1):17291.
[4]王孝友,杨睿,沈克飞,等.兔肺炎克雷伯氏菌的分离与鉴定[J].中国兽医杂志,2012,48(2):43-44.
[5]Kanno H,Katoh N.Analysis by enzyme-linked immunosorbent assay and 2-dimensional electrophoresis of haptoglobin in the high-density lipoprotein fraction in cows[J].J Vet Med Sci,2001,63(1):61-66.
[6]李富祥,李华春,赵德宏,等.肺炎克雷伯氏菌强毒株的分离鉴定及16-23S rRNA ITS序列分析[J].中国畜牧兽医,2015,42(2):352-357.
[7]马磊,颜其贵,万莉,等.竹鼠肺炎克雷伯氏菌的分离鉴定[J].中国人兽共患病学报,2011,27(9):825-827
[8]冯娜,高翔,肖敏,等.牛源肺炎克雷伯氏菌的分离鉴定及遗传进化分析[J].中国兽医科学,2016,46(11):1358-1364.
[9]Jensen M A,Webster J A,Straus N.Rapid identification of bacteria on the basis of polymerase chain reactionamplified ribosomal DNA spacer polymorphisms[J].Appl Environ Microbiol.1993,59(4):945-952.
[10]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[11]金中淦,葛平,徐蓉,等.16S rRNA、16S-23S rRNA基因测序分析检测主要血流感染病原菌比较[J].分子诊断与治疗杂志,2012,4(3):181-185.
[12]Roth A,Reischl U,Streubel A,et al.Novel diagnostic algorithm for identification of mycobacteria using genusspecific amplification of the 16S-23S rRNA gene spacer and restriction endonucleases[J].J Clin Microbiol,2000,38:1094-1104.
[13]Harris K A,Hartley J C.Development of broad-range16S rDNA PCR for use in the routine diagnostic clinical microbiology service[J].J Med Microbiol,2003,52(Pt8):685-691.
[14]于超,郭海勇,魏嘉良,等.16S~23S rRNA基因序列在细菌鉴定中的应用[J].中国畜牧兽医,2012,39(2):57-60.
[15]李瑞蓉,崔雪萍.肺炎克雷伯菌主要耐药机制研究进展[J].中华临床实验室管理电子杂志,2016,4(2):86-90.
[16]周玉龙,李国军,李阳,等.奶牛肺炎克雷伯氏菌的分离与鉴定[J].中国奶牛,2007(6):36-38.
[17]王国卿,王洪海,郭梦尧,等.奶牛产后急性子宫内膜炎病原的分离与鉴定[J].中国兽医杂志,2010,46(10):10-12.