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

Abstract

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.Page 1 of 27A c c e p t e d M a n u s c r i p t A previously isolated contagious ovine digital dermatitis spirochete was located within 34 one of the three phylogroups, group 3, and could also be identified within this group on 35 the basis of phenotype testing, suggesting BDD and contagious ovine digital dermatitis 36 may share the same aetiological agent. A strain isolated from a bovine interdigital 37 dermatitis lesion, could be identified as part of BDD isolate group 2, suggesting bovine 38 interdigital dermatitis and BDD may have the same causative agent. Two common 39 enzyme activities, C4 esterase and C8 esterase lipase, were identified in all BDD 40 associated treponemes suggesting common metabolic pathways for sharing this novel 41 niche or even common virulence traits. Further studies are required to determine whether 42 the three groups of novel treponemes are representative of new treponeme taxa and to 43 delineate how they interact with bovine tissues to cause disease. 44

Association of Unique, Isolated Treponemes with Bovine Digital Dermatitis Lesions

“…Twenty-three strains isolated from BDD lesions and characterized previously were used to validate the PCR tests (13). A previously isolated group 2 strain, G356 (8); a group 3 strain, G179, isolated from a case of contagious ovine DD (CODD) (7); and several reference strains considered to be the closest relatives were also included (complete list in Table 1).…”

“…All biopsies were used for DNA extraction, and 20 of the 29 biopsies were used in an attempt to cultivate treponemes. Biopsies for bacterial cultivation and DNA extraction were collected and transported back to the laboratory as described previously (13), placed immediately in an anaerobic cabinet, and divided into two samples for said procedures. Samples for DNA extraction only were transported on ice without transport medium.…”

“…Strain isolations from BDD lesional tissues, isolate 16S rRNA gene sequencing, and phylogenetic analyses were carried out as previously described (13).…”

“…Cloning of bacterial 16S rRNA genes from BDD lesions in Germany indicated five phylogroups of treponemes present within lesions (5). We recently isolated a large number of treponemes from BDD lesions in the United Kingdom and demonstrated that the isolates belong to three distinct phylogenetic/taxonomic groups (13). We designated the BDD treponemes group 1, group 2, and group 3, which correspond to Treponema medium/Treponema vincentii-like, Treponema phagedenis-like, and Treponema putidum/Treponema denticola-like spirochetes, respectively.…”

Ovine pedomics: the first study of the ovine foot 16S rRNA-based microbiome

“…were detected in only one sheep with ID from flock C. There were two sequences and they had 94 and 99% similarity to uncultured Treponema phylotypes from samples from cattle with digital dermatitis and from animal faecal samples (Klitgaard et al, 2008;Ley et al, 2008). Treponema are often free living but are linked to contagious ovine digital dermatitis and bovine digital dermatitis in sheep and cattle, respectively (Collighan et al, 2000;Moore et al, 2005;Demirkan et al, 2006;Evans et al, 2008Evans et al, , 2009Sayers et al, 2009). Several species have been associated to contagious ovine digital dermatitis and DD including Treponema phagedenis-like and Treponema medium/Treponema vincentii-like, Treponema medium/Treponema vincentii-like, Treponema phagedenis-like, and Treponema denticola/ Treponema putidum-like (Sayers et al, 2009).…”