Association of unique, isolated treponemes with bovine digital dermatitis lesions.

This study used a PCR-based approach targeting 16S rRNA gene fragments to determine the occurrence and association of the three bovine digital dermatitis (BDD) treponeme phylogroups within lesions found in cattle from the United Kingdom. Examination of 51 BDD lesions collected from infected cattle across the United Kingdom revealed that BDD treponeme group 1 (Treponema medium/Treponema vincentii-like), group 2 (Treponema phagedenis-like), and group 3 (Treponema putidum/Treponema denticola-like) were present in 96.1%, 98%, and 76.5% of BDD lesions, respectively. The three phylogroups were present together in 74.5% of lesions. The PCR assays enabled the isolation of further treponeme strains from previously mixed primary BDD lesion cultures. Here a representative from each of the three distinct treponeme phylogroups was isolated from a single BDD lesion for the first time. These data highlight the extent to which this disease is polytreponemal. Immunohistochemistry and electron microscopy were used to investigate lesional hoof tissues, resulting in treponemes being identified copiously in hair follicles and sebaceous glands, suggesting a potential route of exit and/or entry for these pathogens. This study gives further evidence for the importance of the three treponeme groups in BDD pathogenesis and reiterates the value of molecular genetic approaches for isolating and identifying fastidious anaerobes.

In vitro susceptibility of bovine digital dermatitis associated spirochaetes to antimicrobial agents.

Bovine digital dermatitis (BDD) is an infectious lameness in cattle, which has a large global impact in terms of animal welfare and cost. The majority of evidence suggests that spirochaetes are the aetiological agent of this disease. The aim of this study was to identify the susceptibility of BDD associated spirochaetes to a range of antimicrobial agents with a view to potential usage in vivo to treat this widespread cattle disease. A microdilution method was adapted to determine the in vitro susceptibilities of 19 UK digital dermatitis spirochaetes (6 Treponema medium/Treponema vincentii-like, 8 Treponema phagedenis-like and 5 Treponema denticola/Treponema putidum-like) to eight relevant antimicrobials. The BDD spirochaetes exhibited the highest susceptibility to penicillin and erythromycin and this information may now be used to aid development of efficacious treatments. This study has also identified that BDD spirochaete T167 is spectinomycin resistant and that the likely biological basis is a point mutation in the 16S rRNA gene. Interestingly, nearly all Brachyspira isolate 16S rRNA gene sequences in Genbank have this substitution, suggesting it may be responsible for the characteristic spectinomycin resistance reported for the Brachyspira genus.

Three unique groups of spirochetes isolated from digital dermatitis lesions in UK cattle.

Bovine digital dermatitis (BDD) is a severe infectious cause of lameness which has spread through dairy cattle populations worldwide, causing serious welfare and agricultural problems. Spirochetes are the main organisms implicated and have previously proven difficult to isolate. This study aimed to isolate and characterise the range of spirochetes associated with BDD in the UK. Twenty-three spirochete isolates were obtained from 30 BDD lesions, which by 16S rRNA gene and flaB2 gene analysis clustered within the genus Treponema as three phylogroups; groups 1 (Treponema medium/Treponema vincentii-like), 2 (Treponema phagedenis-like) and 3 (Treponema denticola/Treponema putidum-like). The treponemes displayed large genotypic and phenotypic diversity between phylogroups and differed from named treponeme species. A previously isolated contagious ovine digital dermatitis spirochete was located within one of the three phylogroups, group 3, and could also be identified within this group on the basis of phenotype testing, suggesting BDD and contagious ovine digital dermatitis may share the same aetiological agent. A strain isolated from a bovine interdigital dermatitis lesion, could be identified as part of BDD isolate group 2, suggesting bovine interdigital dermatitis and BDD may have the same causative agent. Two common enzyme activities, C4 esterase and C8 esterase lipase, were identified in all BDD associated treponemes suggesting common metabolic pathways for sharing this novel niche or even common virulence traits. Further studies are required to determine whether the three groups of novel treponemes are representative of new treponeme taxa and to delineate how they interact with bovine tissues to cause disease.

Surgical treatment of a sloughed hoof by skin grafting in a young bull.

A sloughed hoof in a young bull as a sequela of foot and mouth disease was treated by skin grafting. Skin blocks obtained from the craniolateral thoracic area of the same animal were placed in slit-like pockets created on the coronary band. New hoof formation was completed within 20 weeks.

Treponema pedis sp. nov., a spirochaete isolated from bovine digital dermatitis lesions.

Bovine digital dermatitis (BDD) is a debilitating infection that is being increasingly recognized in livestock worldwide. Several treponeme phylotypes have been identified in BDD lesions, although only a single BDD-associated treponeme taxon (Treponema brennaborense) has been proposed thus far. In a previous study, we observed that four BDD-associated spirochaete isolates formed a distinct phylogenetic cluster on the basis of 16S rRNA gene sequence analysis and shared less than 97 % sequence similarity with recognized treponeme species. Further characterization of these isolates on the basis of growth characteristics, flaB2 gene sequences, enzyme profiles and cell morphology confirmed that they formed a coherent taxonomic group displaying marked genotypic and phenotypic differences with respect to recognized treponeme species. The four novel isolates displayed a novel 3 : 6 : 3 flagellar pattern rather than the 2 : 4 : 2 pattern shown by their closest relatives and exhibited esterase C4, esterase lipase C8, trypsin and chymotrypsin enzyme activities. Therefore these four new isolates represent a novel species of the genus Treponema, for which the name Treponema pedis sp. nov. is proposed. The type strain is T3552B(T) (=DSM 18691(T)=NCTC 13403(T)).

Reduction and stabilization of hip luxation by the transposition of the ligamentum sacrotuberale in dogs: an in vivo study.

OBJECTIVES: To evaluate the effectiveness of transposition of the sacrotuberous ligament (LST) for the treatment of coxofemoral luxation in the dog. STUDY DESIGN: In vivo experimental study. SAMPLE POPULATION: Ten mixed-breed dogs (weighing 12 to 26 kg). METHODS: After general anesthesia, the LST was exposed and released from the sacrum with a sacral bone fragment. Coxofemoral luxation was created by capsulotomy and transection of the ligament of the head of the femur. Suture was passed through 2 small holes in the bone fragment to guide transposition of the LST through a tunnel drilled through the acetabulum and femoral head and neck. The suture material was pulled tight and an interference screw was placed into the femoral tunnel to lock the sacral bone fragment and LST securely in the tunnel. Butorphanol was administered for pain relief and dogs were allowed unrestricted activity. Coxofemoral radiographs were obtained at 15-day intervals. Two dogs were euthanatized for macroscopic and histopathologic examinations at 3 months. RESULTS: Release of the sacral bone was moderately difficult. The transposed ligament and bone fragment were of an appropriate length and were easily and securely fixed by a interference screw method. Visible severe lameness, during the first 7 to 10 days improved to mild lameness within 10 to 21 days. Gait was subjectively normal after 5 weeks. No radiographic abnormalities were observed at any time point. Grossly, the LST was intact and apparently viable and functional. On histopathology, the space in the bone tunnel was filled with new tissue and a hyaline-like layer surrounded the LST. The sacral bone fragment attached to the LST was united with the femoral bone. CONCLUSIONS: Transposition of the LST reduced and stabilized experimentally induced coxofemoral luxation in 10 dogs. Survival of the LST up to 3 months and its adaptation to transposition suggest that this surgical technique could be considered as a permanent treatment option for hip luxations. Transposition and fixation of the LST was easy, but releasing the sacral edge of the LST was difficult because of its deep location and division of the origin into 2 branches in some dogs. CLINICAL RELEVANCE: Results of this experimental study suggest that the technique may be satisfactorily used in dogs with coxofemoral luxation.