Narayanan S, Nagaraja T G, Okwumabua O, Staats J, Chengappa M M, Oberst R D
Department of Animal Sciences, Kansas State University, Manhattan 66506, USA.
Appl Environ Microbiol. 1997 Dec;63(12):4671-8. doi: 10.1128/aem.63.12.4671-4678.1997.
Restriction fragment length polymorphism analysis of rRNA genes was employed to genetically compare Fusobacterium necrophorum subsp. necrophorum and F. necrophorum subsp. funduliforme isolates from multiple abscesses of the same liver and isolates from liver abscesses, the ruminal wall, and ruminal contents from the same animal. Four livers with multiple abscesses and samples of ruminal contents, ruminal walls, and liver abscesses were collected from 11 cattle at slaughter. F. necrophorum was isolated from all liver abscesses, nine ruminal walls, and six ruminal content samples. Chromosomal DNA of the isolates was extracted and single or double digested with restriction endonucleases (EcoRI, EcoRV, SalI, and HaeIII); then restriction fragments were hybridized with a digoxigenin-labeled cDNA probe transcribed from a mixture of 16S and 23S rRNAs from Escherichia coli. EcoRI alone or in combination with EcoRV yielded the most discriminating ribopatterns for comparison. Within the subspecies multiple isolates from the same liver were indistinguishable based on the ribopattern obtained with EcoRI. The hybridization patterns of liver abscess isolates were concordant with those of the corresponding isolates from ruminal walls in eight of nine sets of samples. None of the six ruminal content isolates matched either the liver abscess isolates or the ruminal wall isolates. The genetic similarity between the isolates from liver abscesses and ruminal walls supports the hypothesis that F. necrophorum isolates of liver abscesses originate from the rumen.
采用rRNA基因的限制性片段长度多态性分析,对来自同一肝脏多个脓肿的坏死梭杆菌坏死亚种和坏死梭杆菌底形亚种分离株,以及来自同一动物肝脏脓肿、瘤胃壁和瘤胃内容物的分离株进行基因比较。在屠宰时从11头牛身上采集了4个有多个脓肿的肝脏以及瘤胃内容物、瘤胃壁和肝脏脓肿的样本。从所有肝脏脓肿、9个瘤胃壁和6个瘤胃内容物样本中分离出坏死梭杆菌。提取分离株的染色体DNA,用限制性内切酶(EcoRI、EcoRV、SalI和HaeIII)进行单酶切或双酶切;然后将限制性片段与用从大肠杆菌16S和23S rRNA混合物转录而来的地高辛标记cDNA探针杂交。单独使用EcoRI或与EcoRV联合使用时,产生的核糖型最具鉴别力,便于比较。在亚种内,基于用EcoRI获得的核糖型,来自同一肝脏的多个分离株无法区分。在9组样本中的8组中,肝脏脓肿分离株的杂交模式与瘤胃壁相应分离株的杂交模式一致。6个瘤胃内容物分离株中没有一个与肝脏脓肿分离株或瘤胃壁分离株匹配。肝脏脓肿分离株和瘤胃壁分离株之间的遗传相似性支持了肝脏脓肿的坏死梭杆菌分离株起源于瘤胃的假说。