Digital Dermatitis Lesions Research Articles

Characterization of Treponema phagedenis-like spirochetes isolated from papillomatous digital dermatitis lesions in dairy cattle.

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.

Genetic relatedness and phenotypic characteristics of Treponema associated with human periodontal tissues and ruminant foot disease.

Treponema have been implicated recently in the pathogenesis of digital dermatitis (DD) and contagious ovine digital dermatitis (CODD) that are infectious diseases of bovine and ovine foot tissues, respectively. Previous analyses of treponemal 16S rDNA sequences, PCR-amplified directly from DD or CODD lesions, have suggested relatedness of animal Treponema to some human oral Treponema species isolated from periodontal tissues. In this study a range of adhesion and virulence-related properties of three animal Treponema isolates have been compared with representative human oral strains of Treponema denticola and Treponema vincentii. In adhesion assays using biotinylated treponemal cells, T. denticola cells bound in consistently higher numbers to fibronectin, laminin, collagen type I, gelatin, keratin and lactoferrin than did T. vincentii or animal Treponema isolates. However, animal DD strains adhered to fibrinogen at equivalent or greater levels than T. denticola. All Treponema strains bound to the amino-terminal heparin I/fibrin I domain of fibronectin. 16S rDNA sequence analyses placed ovine strain UB1090 and bovine strain UB1467 within a cluster that was phylogenetically related to T. vincentii, while ovine strain UB1466 appeared more closely related to T. denticola. These observations correlated with phenotypic properties. Thus, T. denticola ATCC 35405, GM-1, and Treponema UB1466 had similar outer-membrane protein profiles, produced chymotrypsin-like protease (CTLP), trypsin-like protease and high levels of proline iminopeptidase, and co-aggregated with human oral bacteria Porphyromonas gingivalis and Streptococcus crista. Conversely, T. vincentii ATCC 35580, D2A-2, and animal strains UB1090 and UB1467 did not express CTLP or trypsin-like protease and did not co-aggregate with P. gingivalis or S. crista. Taken collectively, these results suggest that human oral-related Treponema have broad host specificity and that similar control or preventive strategies might be developed for human and animal Treponema-associated infections.

From tooth to hoof: treponemes in tissue-destructive diseases.

With the advent of new molecular and immunological tools, there is better understanding of the roles that difficult to cultivate bacteria, and not-yet-cultivated bacteria such as spirochaetes, play in polymicrobial diseases. Only relatively recently have studies implicated Treponema spirochaetes in human periodontal disease, a destructive condition of the tissues supporting the teeth. A number of different Treponema species have been isolated and their surface protein components that mediate adhesion, cytotoxicity, and tissue damage have been characterized. More recently Treponema strains closely related to human oral isolates have been cultivated from active lesions of digital dermatitis, an ulcerative condition affecting the feet of cows and sheep. This condition, like periodontal disease, appears to have a polymicrobial aetiology in which enrichment for Treponema may play a crucial part. This article reviews the known mechanisms by which Treponema interact with eukaryotic host cells and tissue proteins, and how these interactions may contribute to pathogenic diversity.

Efficacy of Two Modified Nonantibiotic Formulations (Victory) for Treatment of Papillomatous Digital Dermatitis in Dairy Cows

A field trial was conducted to compare the efficacy of the original and two modified formulations of Victory and oxytetracycline among dairy cows affected with papillomatous digital dermatitis. Seventy-eight cows with papillomatous digital dermatitis lesions were randomly allocated to one of four treatment groups (A, B, C, D). Cows in group A (n = 19) were treated with an oxytetracycline solution; cows in group B (n = 22) were treated with the original formulation of Victory containing soluble copper, peroxide compound, and a cationic agent; cows in group C (n = 17) were treated with a modified formulation of Victory containing reduced soluble copper and peroxide compound but increased levels of cationic agent; and cows in group D (n = 20) were treated with a modified formulation of Victory containing levels of soluble copper and cationic agent equivalent to the original formulation but with reduced concentrations of peroxide compound. Cows were examined 7, 14 and 28 d after initial treatment; during each examination, pain and lesion scores were recorded. The modified nonantibiotic formulation used in cows in group C appeared to be the most effective for treatment of papillomatous digital dermatitis. Proportions of cows with signs of pain were significantly lower among cows in group C, compared with cows in group A. Similarly, pain scores were significantly lower among cows in treatment group C, compared to cows in group A. The low efficacy of oxytetracycline was an unexpected result and may have clinical implications associated with possible antibiotic resistance in dairy cows affected with papillomatous digital dermatitis.

Clinical Effect of Local Antibiotic Treatment of Digital Dermatitis Lesions and Detection of Spirochetes by Means of PCR

The etiology of digital dermatitis (DD) remains to be fully understood. The disease has been associated with the presence of anaerobic bacteria, though, which most likely are spirochetes (Walker et al., 1997).
 The purpose of the study was to monitor the clinical healing process and the recurrence of lesions after topic antibiotic spray treatment. The presence of spirochetes in superficial wound smears taken during the course of the disease was monitored by polymerase chain reaction (PCR).

Uniform protocol for evaluating response to treatment of papillomatous digital dermatitis lesions

Digital dermatitis or papillomatous digital dermatitis (PDD), commonly known as footwarts or hairy heel warts, was first reported in Italy in 197 4 and in New York in the late 1970's. Similar lesions have occurred in 40 or more states in the United States and many other countries. The etiology of this disease is unknown, however, two to five phylotypes of spirochetes (Treponema sp.) have been associated with active lesions and identified by using Polymerase Chain Reaction (PCR), histologic, or microbiological methods.
 The purpose of this paper is to develop a uniform protocol which can be used to evaluate results of treatment to control PDD lesions. The remainder of this paper lists our suggestions for information which should be included in each trial protocol.

Treponema brennaborense sp. nov., a novel spirochaete isolated from a dairy cow suffering from digital dermatitis.

A novel Treponema species was isolated from an ulcerative lesion of a cow suffering from digital dermatitis (DD), a disease which causes painful ulcerations along the coronary band. Among other anaerobic bacteria, high numbers of spirochaetes have been regularly found in DD lesions. Here data are presented of a spirochaete isolated from a DD ulcer. By chemotaxonomy, protein analysis and comparative 16S rDNA sequence analysis this isolate was classified as a treponeme that differed from all Treponema species described previously. The only isolate, DD5/3T, for which the name Treponema brennaborense is proposed, is designated the type strain of the novel species. The strain is a small, highly motile spirochaete that has two periplasmic flagella, one flagellum being attached at each cell pole. Strain DD5/3T exhibits alpha-glucosidase and N-acetyl-beta-glucosaminidase activity and growth is inhibited by rabbit serum. T. brennaborense was phylogenetically most closely related (89.5% 16S rRNA similarity) to Treponema maltophilum, an oral spirochaete isolated from a periodontitis patient.

Efficacy of Serpens spp bacterin combined with topical administration of lincomycin hydrochloride for treatment of papillomatous digital dermatitis (footwarts) in cows on a dairy in California

The objective of this prospective, randomized field trial was to determine if a Serpens spp bacterin had a therapeutic effect on cows with active lesions of papillomatous digital dermatitis (PDD) before or in combination with treatment with topical lincomycin HCl on a California dairy. Fifty-nine lactating cows with active, painful PDD were randomly assigned to control (not vaccinated) and vaccinated groups. Lesions of the rear feet were evaluated for pain response, color, size, and lesion scores at each of the examinations. Cows in the vaccinated group were vaccinated 3 times with a Serpens spp bacterin (days 1, 14, 43). All cows were restrained on a hydraulic tilt table at approximately day 30 and, after examination, all lesions were treated with a slurry containing 3.2 g of lincomycin mixed with 3 ml of deionized water. The slurry was applied to cotton gauze and held in place with an elastic bandage. Bandages were allowed to wear off. Lesions were re-examined while cows were locked in stanchions at approximately days 50 and 70 and while restrained on the hydraulic tilt table on approximately day 110. Lesions improved on all cows during the course of the trial. Lesion and size scores were improved for control and vaccinated cows on days 50, 70, and 110. Pain response and color scores improved on days 50 and 70 but showed evidence of recurrence or recrudescence by day 110. There were no significant differences in evaluated scores between control and vaccinated cows at any of the examination times. When affected cows were re-examined at day 110, recurrence or recrudescence of lesions was evident in 41% of the control cows and 45% of the vaccinated cows. Vaccination of PDD affected cows with Serpens spp bacterin had no apparent effect on clinical outcome.

Fluorescence in situ hybridization shows spatial distribution of as yet uncultured treponemes in biopsies from digital dermatitis lesions.

Fluorescence in situ hybridization (FISH) was performed on sections of plastic-embedded tissue using 16S rRNA-directed oligonucleotide probes to visualize uncultured treponemes in skin biopsies of cows with digital dermatitis. Plastic as embedding material allowed sectioning of hard and soft tissue with a defined thickness, avoiding the risk of dragging bacteria into the tissue while sectioning. furthermore, it provided a good signal-to-noise ratio. Using this method the spatial distribution of three different bacterial phylotypes was visualized simultaneously within the tissue. Whereas debris covering the ulcers contained a mixture of different micro-organisms, a layering of certain treponemal phylotypes was observed deeper in the epidermis. Confocal laser scanning microscopy and subsequent three-dimensional reconstruction of series of optical sections confirmed that the treponemes migrated intercellularly around the cells, most of them directed towards the dermis. In situ hybridization on tissue embedded in plastic proved to be a useful method to study mixed bacterial infections since it combines excellent histological conservation of tissue with identification of bacterial species by simultaneous use of probes labelled with different fluorescent dyes. This technique may have implications for in situ detection, identification and localization of microorganisms in veterinary as well as in human medicine.

Spirochaetes and other bacterial species associated with bovine digital dermatitis

The 16S rRNA genes from spirochaetes associated with digital dermatitis of British cattle were amplified by polymerase chain reaction from digital dermatitis lesion biopsies using one universal and one treponeme-specific primer. Two treponemal sequences were identified both of which shared a high degree of homology with the oral pathogen Treponema denticola (98%). Two further 16S rRNA gene sequences were obtained and shared similarity to Bacteroides levii (99%) and Mycoplasma hyopharyngis (98%). Polymerase chain reaction with T. denticola-specific primers amplified a potential virulence gene from digital dermatitis lesions which shared a high degree of homology to the 46-kDa haemolysin gene of T. denticola. The significance of the presence of organisms in digital dermatitis lesions of the bovine foot which are closely related to oral pathogens is discussed.

Humoral response of dairy cattle to spirochetes isolated from papillomatous digital dermatitis lesions

Abstract Objective To determine whether a humoral response against spirochetes isolated from papillomatous digital dermatitis (PDD) lesions is elicited in dairy cattle affected with PDD. Sample Population 41 cattle with PDD from 8 dairies (study population) and 30 cattle from 2 dairies free of PDD (control population). Additionally evaluated were 32 cattle from a dairy with a past history of PDD but no current disease, and 52 cattle from a dairy with high prevalence of PDD, 25 with and 27 without detectable lesions. Procedure ELISA were used to evaluate the humoral response of all cattle to representative isolates from 2 groups of spirochetes of unknown species isolated from PDD lesions. Specificity of the response was evaluated, using immune sera prepared against each of the spirochetes, and by adsorption studies of immune and field sera. The potential for confounding by an antibody response to other spirochetes associated with diseases of cattle was assessed. Results The antibody response (specific) to both PDD spirochete groups of cows with PDD was significantly increased, compared with that of cows from PDD-free dairies. There was no association between antibody response to PDD-associated spirochetes and antibody response to other spirochetal diseases of cattle. None of the cattle from the dairy with previous history of PDD but without current disease were classified as test positive by either PDD ELISA. There was a significant (P < 0.01) difference in classification results for both PDD ELISA for cattle with PDD from the dairy with a high herd prevalence of PDD, compared with cattle without detectable disease from the same dairy. Conclusions and Clinical Relevance The humoral response in cattle with PDD lesions was significantly different from that in cattle without detectable lesions, thus providing additional information regarding the potential role of spirochetes isolated from PDD lesions in the etiopathogenesis of PDD. (Am J Vet Res 1997;58:744–748)

Partial identification of spirochaetes from two dairy cows with digital dermatitis by polymerase chain reaction analysis of the 16S ribosomal RNA gene

Specimens taken postmortem from typical lesions of digital dermatitis in two dairy cows were tested by the polymerase chain reaction (PCR) for the presence of a spirochaetal 16S rRNA gene....

Spirochetes from digital dermatitis lesions in cattle are closely related to treponemes associated with human periodontitis.

Digital dermatitis (DD), first described in 1974 by Cheli and Mortellaro (R. Cheli and C. Mortellaro, p. 208-213, in Proceedings of the 8th International Conference on Diseases of Cattle, 1974), is a major problem in diary cows and beef cattle causing significant economic losses worldwide. Lesions are typically found at the volar skin proximal to the heel bulbs. Microscopic examination of biopsies or touch preparations of these lesions revealed a variety of different bacterial morphotypes including significant numbers of spirochetes which often represent the predominant morphotype. We used comparative 16S rRNA sequence analysis to determine the diversity and phylogeny of these hitherto unclassified DD spirochetes. Results indicate that those lesions looked at so far contained at least five spirochetal phylotypes, all clustering within the genus Treponema. Phylotype DDKL-4 was nearly identical (99.4% similarity) to that of a nonpathogenic human treponeme, T. phagedenis. Two phylotypes DDKL-3 and DDKL-13 were closely related to those from treponemes commonly found in human periodontitis lesions, i.e., T. denticola and T. vincetii, exhibiting 95 and 98% similarity, respectively. The other two phylotypes, DDKL-12 and DDKL-20, had no close relatives to any cultivable treponemal species but clustered to previously described group IV oral treponemes. Preliminary analysis using in situ hybridization with fluorescently labeled oligonucleotide probes against smears from DD biopsies revealed that from all lesions analyzed so far, T. denticola-like spirochetes were detected in the highest proportion of all spirochetal morphotypes.

Spirochetes isolated from dairy cattle with papillomatous digital dermatitis and interdigital dermatitis

Two groups of spirochetes were isolated from papillomatous digital dermatitis (PDD) lesions in dairy cattle. The two groups could be readily differentiated on the basis of morphologic and immunologic characteristics and enzymatic activity. A spirochete isolated from an interdigital dermatitis (IDD) lesion appeared morphologically and antigenically similar to spirochetes in one of the PDD groups and exhibited an identical enzyme activity pattern. The two groups of PDD spirochetes had characteristics most consistent with the genus Treponema. The PDD and IDD isolates differed morphologically from previously described bovine Treponema spp. Although spirochetes have been observed to be one of the predominant bacterial morphotypes in PDD and IDD and are found invading the stratum spinosum and dermal papillae in PDD lesions, the significance of these spirochetes in the etiopathogenesis of PDD and IDD is presently unknown.

Evaluation of Four Therapies of Papillomatous Digital Dermatitis in Dairy Cattle

One hundred and forty-seven Holstein cows in three Central California dairy herds, housed in open corrals with shades, with active Papillomatous Digital Dermatitis (hairy footwart) lesions were randomly assigned to one of five treatment groups.