Characterization of Treponema phagedenis -Like Spirochetes Isolated from Papillomatous Digital Dermatitis Lesions in Dairy Cattle

Abstract

Four spirochete strains were isolated from papillomatous digital dermatitis (PDD) lesions in Iowa dairy cattle and compared with two previously described spirochete strains isolated from dairy cattle in California. These six strains shared an identical 16S ribosomal DNA sequence that was 98% similar to Treponema phagedenis and 99% similar to the uncultivated PDD spirochete sequence DDLK-4. The whole-cell protein profiles resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of these six strains were similar. However, these strains showed differences in the antigenic diversity of lipopolysaccharide (LPS). Genetic diversity was also detected by pulsed-field gel electrophoresis of genomic DNA digests, revealing differences among five of the six strains. Serum immunoglobulin G antibodies from dairy cattle with active PDD lesions reacted with the LPS of all but one PDD spirochete strain. Likewise, peripheral blood mononuclear cells from cattle with active PDD lesions produced blastogenic responses to one of the two California isolates. Both antibody and lymphocyte blastogenic responses were reduced in convalescent dairy cattle, suggesting the immune response to these spirochetes has short duration. These results demonstrate genetic and antigenic diversity among T. phagedenis-like treponemes and provide further evidence for the involvement of these spirochetes in the pathogenesis of PDD.Papillomatous digital dermatitis (PDD) is a leading cause of lameness in dairy cattle (23). This disease state is referred to by a variety of names including digital dermatitis, interdigital papillomatosis, hairy heelwart, and hairy footwart. PDD begins as a mild, superficial dermatitis which progresses to an erosive lesion. The tissue becomes granulated and can form a hyperkeratotic papillomatous lesion with long hair-like projections (3,29,30). These lesions usually form on the plantar surface, proximal to the bulb of the heel, or occasionally, within the interdigital cleft, and are thought to arise due to constant exposure to fecal slurry (2, 28).The first report of PDD was in 1974 in Europe (8), and the disease is now found with increasing frequency in most geographical regions of the world. In the United States, the disease was identified in 43% of 1,182 U.S. dairy herds surveyed in 1996 and 78% of these herds reported their first cases in 1993 or later. These data confirm a rapid spread of the disease (46), which may in part be due to changes in herd management (31,32,46).The etiology of PDD remains equivocal. Application of topical or parenteral antibiotics results in rapid resolution of the lesions, suggesting that PDD is caused by a bacterial infection. A mixed population of gram-negative bacteria, including anaerobes, microaerophilic organisms, and spirochetes have been demonstrated in or isolated from PDD lesions (4,26,29).Spirochetes are prevalent in PDD lesions and may be important in pathogenesis. Spirochetes are found deep within the epidermis, invading the stratum spinosum and dermal papillae (2,4,22,29)...

Association of Unique, Isolated Treponemes with Bovine Digital Dermatitis Lesions

“…This indicated the presence of two (or, most commonly, three) phylogroups in BDD lesions. Isolation data from previous studies suggested that the group 2 isolates (T. phagedenis-like) might be the only phylogroup specifically associated with BDD, as they have been the most commonly isolated ones (13,32,33). The current data clearly refute this premise, as the new PCR tests indicated the presence of group 2 treponemes in nearly all BDD lesions (98%) and detected group 1 treponemes (T. medium/T.…”

Evidence of Multiple Treponema Phylotypes Involved in Bovine Digital Dermatitis as Shown by 16S rRNA Gene Analysis and Fluorescence In Situ Hybridization

“…Still, the high number of invasive spirochetes observed suggests an active contribution of this group to DD pathogenesis (8,9,12,16,24,35). This hypothesis is further substantiated by the fact that serum samples from cattle infected with DD contain elevated levels of antibody to Treponema antigens (13,25,36). Also, it has been demonstrated that four Treponema strains previously isolated from DD-infected cows were able to induce abscess formation in mice (18).…”

Autoreactive T cells specific for insulin B:11-23 recognize a low-affinity peptide register in human subjects with autoimmune diabetes

“…We identified a number of environmental microbes expressing proteins that contain peptide sequences with high homology to the R3 core region of B:11-23 R22E peptide (ALYLVCGEE). These microbes included species that are commonly isolated from poultry or dairy animals and environmental microbes that are commonly encountered (23)(24)(25)(26). DQ8-restricted insulin-specific T-cell clones were activated by these homologous microbial peptides as effectively as the B:11-23 R22E peptide.…”

“…We identified four microbial peptides with high-sequence homology to ALYLVCGEE (Table S3). These sequences were derived from Bacteroides salanitronis, a common inhabitant of chicken intestine/cecum (23), Clostridium acetobutylicum, a soil dwelling Clostridium species (24), Treponema phagedenis, a pathogen of papillomatous digital dermatitis isolated from Iowa dairy cattle (25), and Microscilla marina, a bacterial group frequently found on marine particulates (26). All of the T-cell clones tested responded to stimulation with these bacterial peptides and the responses, as evaluated by the stimulation index, were of comparable magnitude to the modified B:11-23 R22E peptide (Fig.…”