Muzafar Mohd, Calvo-Bado Leo A, Green Laura E, Smith Edward M, Russell Claire L, Grogono-Thomas Rose, Wellington Elizabeth M H
School of Life Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.
Department of Clinical Veterinary Sciences, University of Bristol, Langford House, Langford BS40 5DU, UK.
Vet Microbiol. 2015 Aug 31;179(1-2):53-9. doi: 10.1016/j.vetmic.2015.04.010. Epub 2015 Apr 22.
Dichelobacter nodosus (D. nodosus) is the essential causative agent of footrot in sheep. The current study investigated when D. nodosus was detectable on newborn lambs and possible routes of transmission. Specific qPCR was used to detect and quantify the load of D. nodosus in foot swabs of lambs at birth and 5-13 h post-partum, and their mothers 5-13 h post-partum; and in samples of bedding, pasture, soil and faeces. D. nodosus was not detected on the feet of newborn lambs swabbed at birth, but was detected 5-13 h after birth, once they had stood on bedding containing naturally occurring D. nodosus. Multiple genotypes identified by cloning and sequencing a marker gene, pgrA, and by multi locus variable number tandem repeat analysis (MLVA) of community DNA from swabs on individual feet indicated a mixed population of D. nodosus was present on the feet of both ewes and lambs. There was high variation in pgrA tandem repeat number (between 3 and 21 repeats), and multiple MLVA types. The overall similarity index between the populations on ewes and lambs was 0.45, indicating moderate overlap. Mother offspring pairs shared some alleles but not all, suggesting lambs were infected from sources(s) other than just their mother's feet. We hypothesise that D. nodosus is transferred to the feet of lambs via bedding containing naturally occurring populations of D. nodosus, probably as a result of transfer from the feet of the group of housed ewes. The results support the hypothesis that the environment plays a key role in the transmission of D. nodosus between ewes and lambs.
坏死梭杆菌是绵羊腐蹄病的主要致病因子。本研究调查了新生羔羊何时可检测到坏死梭杆菌以及可能的传播途径。采用特异性定量聚合酶链反应(qPCR)检测并定量出生时及产后5 - 13小时羔羊蹄拭子、产后5 - 13小时母羊蹄拭子,以及垫料、牧场、土壤和粪便样本中坏死梭杆菌的载量。出生时采集的新生羔羊蹄拭子未检测到坏死梭杆菌,但出生后5 - 13小时,一旦它们站立在含有自然存在的坏死梭杆菌的垫料上,就检测到了该菌。通过克隆和测序标记基因pgrA以及对单个蹄拭子的群落DNA进行多位点可变数目串联重复分析(MLVA)鉴定出的多种基因型表明,母羊和羔羊的蹄上均存在坏死梭杆菌混合菌群。pgrA串联重复数存在很大差异(3至21个重复之间),且有多种MLVA类型。母羊和羔羊群体之间的总体相似性指数为0.45,表明有中度重叠。母羊与后代对共享一些等位基因,但并非全部,这表明羔羊感染的来源不止其母羊的蹄部。我们推测,坏死梭杆菌通过含有自然存在的坏死梭杆菌菌群的垫料转移到羔羊蹄部,可能是由于圈养母羊群蹄部的传播所致。这些结果支持了环境在坏死梭杆菌在母羊和羔羊之间传播中起关键作用这一假说。