Abstract
BackgroundDigital dermatitis in cattle is an emerging infectious disease. Ulcerative lesions are typically located on the plantar skin between the heel bulbs and adjacent to the coronet. Spirochetes of the genus Treponema are found in high numbers in the lesions and are likely to be involved in the pathogenesis. The aim of this study was to obtain pure cultures of spirochetes from cattle with digital dermatitis and to describe them further.MethodsTissue samples and swabs from active digital dermatitis lesions were used for culturing. Pure isolates were subjected to, molecular typing through 16S rRNA gene sequencing, pulsed-field gel electrophoresis (PFGE), random amplified polymorphic DNA (RAPD) and an intergenic spacer PCR developed for Treponema spp. as well as API-ZYM and antimicrobial susceptibility tests. The antimicrobial agents used were tiamulin, valnemulin, tylosin, aivlosin, lincomycin and doxycycline.ResultsSeven spirochete isolates from five herds were obtained. Both 16S rRNA gene sequences, which were identical except for three polymorphic nucleotide positions, and the intergenic spacer PCR indicated that all isolates were of one yet unnamed species, most closely related to Treponema phagedenis. The enzymatic profile and antimicrobial susceptibility pattern were also similar for all isolates. However it was possible to separate the isolates through their PFGE and RAPD banding pattern.ConclusionThis is the first report on isolation of a Treponema sp. from cattle with digital dermatitis in Scandinavia. The phylotype isolated has previously been cultured from samples from cattle in the USA and the UK and is closely related to T. phagedenis. While very similar, the isolates in this study were possible to differentiate through PFGE and RAPD indicating that these methods are suitable for subtyping of this phylotype. No antimicrobial resistance could be detected among the tested isolates.
Highlights
Digital dermatitis in cattle is an emerging infectious disease
Raw sequence data without any traces of contamination, using non species specific Treponema primers both for the 16S rRNA gene and the intergenic spacer, indicate that > 95% of the cultures were of one species
When different DNA preparations from the same isolate were analysed by pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD), identical banding patterns indicate that the cultures contained a single species
Summary
Digital dermatitis in cattle is an emerging infectious disease. Ulcerative lesions are typically located on the plantar skin between the heel bulbs and adjacent to the coronet. Spirochetes of the genus Treponema are found in high numbers in the lesions and are likely to be involved in the pathogenesis. The aim of this study was to obtain pure cultures of spirochetes from cattle with digital dermatitis and to describe them further. Digital dermatitis (DD) is a contagious claw disease causing lameness in cattle, most commonly seen in intensive dairy production. The disease was first described in 1974 in Italy by Cheli and Mortellaro [1]. There is a strong connection between wet/dirty claw environments and the occurrence of DD [4], for example in cubicle systems where accumulation of faeces and urine on the alleys is a typical hygienic problem. Besides being an animal welfare problem, economic losses due to reduced milk production and weight loss are associated with DD [5]
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