De Groote D, van Doorn L J, Ducatelle R, Verschuuren A, Tilmant K, Quint W G, Haesebrouck F, Vandamme P
Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, University of Ghent, Merelbeke, Belgium.
Int J Syst Bacteriol. 1999 Oct;49 Pt 4:1707-15. doi: 10.1099/00207713-49-4-1707.
Recently helicobacter-like organisms have been reported in the pyloric part of the abomasum of calves and adult cattle. Cultivation of these spiral bacteria has not been successful to date. In the present study, comparative 16S rDNA sequence analysis was used to determine the taxonomic position of these bacteria. Seven abomasal biopsies of adult cattle were sampled from different Belgian and Dutch farms. In all samples the presence of helicobacter-like organisms was demonstrated by biochemical, immunohistochemical and electron microscopical data. Bacterial 16S rDNA was amplified by PCR and sequences were determined either by direct or indirect sequence analysis. Pairwise comparisons revealed all sequences to be more than 99% homologous. Phylogenetic analysis placed the organism, corresponding to the reference sequence R2XA, within the genus Helicobacter. A diagnostic PCR assay was designed, differentiating all of the bovine 16S rDNA sequences from Helicobacter and Wolinella species. The low similarity level towards Helicobacter bilis (92.8%), its closest validly named neighbour, indicates that this novel taxon is indeed a novel Helicobacter species. An in situ hybridization procedure associated the bovine sequences to the helicobacter-like organisms in the abomasum. The name 'Candidatus Helicobacter bovis' is proposed for this new abomasal helicobacter from cattle.
最近，在犊牛和成年牛皱胃的幽门部发现了螺旋杆菌样微生物。迄今为止，这些螺旋菌的培养尚未成功。在本研究中，采用比较16S rDNA序列分析来确定这些细菌的分类地位。从比利时和荷兰不同农场采集了7份成年牛皱胃活检样本。通过生化、免疫组织化学和电子显微镜数据证实，所有样本中均存在螺旋杆菌样微生物。通过PCR扩增细菌16S rDNA，并通过直接或间接序列分析确定序列。成对比较显示所有序列的同源性超过99%。系统发育分析将与参考序列R2XA相对应的微生物置于螺旋杆菌属内。设计了一种诊断性PCR检测方法，可区分所有牛16S rDNA序列与螺旋杆菌属和沃氏菌属物种。与最接近的有效命名邻居胆汁螺旋杆菌的相似性水平较低（92.8%），表明这个新分类单元确实是一个新的螺旋杆菌物种。原位杂交程序将牛序列与皱胃中的螺旋杆菌样微生物联系起来。建议将这种来自牛的新的皱胃螺旋杆菌命名为“暂定牛螺旋杆菌”。
