Department of Biological Sciences, School of Life Sciences, University of Warwick, Coventry, UK.
ISME J. 2011 Sep;5(9):1426-37. doi: 10.1038/ismej.2011.25. Epub 2011 Mar 24.
We report the first study of the bacterial microbiome of ovine interdigital skin based on 16S rRNA by pyrosequencing and conventional cloning with Sanger-sequencing. Three flocks were selected, one a flock with no signs of footrot or interdigital dermatitis, a second flock with interdigital dermatitis alone and a third flock with both interdigital dermatitis and footrot. The sheep were classified as having either healthy interdigital skin (H) and interdigital dermatitis (ID) or virulent footrot (VFR). The ovine interdigital skin bacterial community varied significantly by flock and clinical condition. The diversity and richness of operational taxonomic units was greater in tissue from sheep with ID than H or VFR-affected sheep. Actinobacteria, Bacteriodetes, Firmicutes and Proteobacteria were the most abundant phyla comprising 25 genera. Peptostreptococcus, Corynebacterium and Staphylococcus were associated with H, ID and VFR, respectively. Sequences of Dichelobacter nodosus, the causal agent of ovine footrot, were not amplified because of mismatches in the 16S rRNA universal forward primer (27F). A specific real-time PCR assay was used to demonstrate the presence of D. nodosus, which was detected in all samples including the flock with no signs of ID or VFR. Sheep with ID had significantly higher numbers of D. nodosus (10(4)-10(9) cells per g tissue) than those with H or VFR feet.
我们报告了首个基于 16S rRNA 的焦磷酸测序和传统克隆 Sanger 测序的绵羊趾间皮肤细菌宏基因组研究。选择了三个羊群,一个没有腐蹄病或趾间皮炎迹象的羊群,一个只有趾间皮炎的羊群,一个既有趾间皮炎又有腐蹄病的羊群。绵羊被分为健康趾间皮肤(H)和趾间皮炎(ID)或毒力腐蹄病(VFR)。绵羊趾间皮肤细菌群落因羊群和临床状况而异。与 H 或 VFR 受影响的绵羊相比,ID 绵羊的组织中操作分类单位的多样性和丰富度更高。放线菌、拟杆菌门、厚壁菌门和变形菌门是最丰富的门,包含 25 个属。消化链球菌属、棒状杆菌属和葡萄球菌属分别与 H、ID 和 VFR 相关。由于 16S rRNA 通用正向引物(27F)中的错配,未能扩增腐蹄病的病原体 Dichelobacter nodosus 的序列。使用特定的实时 PCR 检测法证明了 D. nodosus 的存在,该菌存在于所有样本中,包括没有 ID 或 VFR 迹象的羊群。患有 ID 的绵羊的 D. nodosus 数量明显高于 H 或 VFR 脚的绵羊(每克组织 10(4)-10(9)个细胞)。