Targeting the Treponemal Microbiome of Digital Dermatitis Infections by High-Resolution Phylogenetic Analyses and Comparison with Fluorescent
            <i>In Situ</i>
            Hybridization

Klitgaard, Kirstine; Bretó, Antoni Foix; Boye, Mette; Jensen, Tim Kåre

doi:10.1128/jcm.00320-13

Cited by 59 publications

(84 citation statements)

References 35 publications

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“…Moreover, a range of metagenomic studies have identified the association of specific treponeme phylogroups with DD lesions in Europe, Japan, and the United States (31)(32)(33)(34). In each of these studies, other bacterial genera were identified in DD lesions; however, only for the treponemes were there strong association data across all these studies.…”

Section: Discussionmentioning

confidence: 93%

Isolation of Digital Dermatitis Treponemes from Hoof Lesions in Wild North American Elk (Cervus elaphus) in Washington State, USA

et al. 2015

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c Since 2008, a large increase in the numbers of cases of lameness have been seen in wild North American elk (Cervus elaphus) from Washington State, USA. The most recent cases manifested as foot lesions similar both clinically and pathologically to those seen in digital dermatitis (DD) in cattle and sheep, a disease with a bacterial etiopathogenesis. To determine whether the same bacteria considered responsible for DD are associated with elk lameness, lesion samples were subjected to bacterial isolation studies and PCR assays for three phylogroups of relevant DD treponemes. The DD treponemes were isolated from lesional tissues but not from control feet or other areas of the diseased foot (including the coronary band or interdigital space), suggesting that the bacteria are strongly associated with DD lesions and may therefore be causal. In addition, PCR analysis revealed that all three unique DD treponeme phylotypes were found in elk hoof disease, and in 23% of samples, all 3 DD-associated treponemes were present in lesions. Sequence analysis of the 16S rRNA gene showed that the elk lesion treponemes were phylogenetically almost identical to those isolated from cattle and sheep DD lesions. The isolates were particularly similar to two of the three culturable DD treponeme phylotypes: specifically, the Treponema medium/Treponema vincentii-like and Treponema phagedenislike DD spirochetes. The third treponeme culturable phylogroup (Treponema pedis), although detected by PCR, was not isolated. This is the first report describing isolation of DD treponemes from a wildlife host, suggesting that the disease may be evolving to include a wider spectrum of cloven-hoofed animals.

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Section: Discussionmentioning

confidence: 93%

Isolation of Digital Dermatitis Treponemes from Hoof Lesions in Wild North American Elk (Cervus elaphus) in Washington State, USA

et al. 2015

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“…also appears to have different constituents at each of the various stages of development, which may help explain the variability noted in the Treponema spp. visualized from previous fluorescent in situ hybridization studies (33,34,54). The significance of these changes in Treponema species and relative abundance in relation to the possible role of Treponema spp.…”

Section: Discussionmentioning

confidence: 99%

“…Several researchers (15,53) have used 16S-based cloning to evaluate the DD microbiome by classic Sanger sequencing, creating at most 1,525 clones from nine bovine feet. Klitgaard et al (54) utilized high-throughput 16S-based 454 sequencing to create 212,827 sequences from 40 cows; however, instead of using universal 16S primers, they utilized a custom Treponema-specific primer to explore the diversity of this bacterial genus in each biopsy specimen (54). Consequently, the relative abundance of the Treponema spp.…”

Section: Discussionmentioning

confidence: 99%

Deep Sequencing Analysis Reveals Temporal Microbiota Changes Associated with Development of Bovine Digital Dermatitis

et al. 2014

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bBovine digital dermatitis (DD) is a leading cause of lameness in dairy cattle throughout the world. Despite 35 years of research, the definitive etiologic agent associated with the disease process is still unknown. Previous studies have demonstrated that multiple bacterial species are associated with lesions, with spirochetes being the most reliably identified organism. This study details the deep sequencing-based metagenomic evaluation of 48 staged DD biopsy specimens collected during a 3-year longitudinal study of disease progression. Over 175 million sequences were evaluated by utilizing both shotgun and 16S metagenomic techniques. Based on the shotgun sequencing results, there was no evidence of a fungal or DNA viral etiology. The bacterial microbiota of biopsy specimens progresses through a systematic series of changes that correlate with the novel morphological lesion scoring system developed as part of this project. This scoring system was validated, as the microbiota of each stage was statistically significantly different from those of other stages (P < 0.001). The microbiota of control biopsy specimens were the most diverse and became less diverse as lesions developed. Although Treponema spp. predominated in the advanced lesions, they were in relatively low abundance in the newly described early lesions that are associated with the initiation of the disease process. The consortium of Treponema spp. identified at the onset of disease changes considerably as the lesions progress through the morphological stages identified. The results of this study support the hypothesis that DD is a polybacterial disease process and provide unique insights into the temporal changes in bacterial populations throughout lesion development.

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“…The bacteria most often associated with DD are spirochetes of the genus Treponema, which predominate in the deepest regions of DD lesions (7,8). To date, at least 20 different phylotypes have been identified from DD biopsy specimens; among these, Treponema phagedenis-like, Treponema medium/Treponema vincentii-like, Treponema denticolalike, and Treponema pedis phylotypes are highly associated with progression of the disease (9)(10)(11)(12). DD also appears to be highly infectious based on the rapid intra-and interherd spread of the disease (6,13,14).…”

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confidence: 99%

Discovery of Bovine Digital Dermatitis-Associated Treponema spp. in the Dairy Herd Environment by a Targeted Deep-Sequencing Approach

Klitgaard

Nielsen

Ingerslev

et al. 2014

Appl Environ Microbiol

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The bacteria associated with the infectious claw disease bovine digital dermatitis (DD) are spirochetes of the genus Treponema; however, their environmental reservoir remains unknown. To our knowledge, the current study is the first report of the discovery and phylogenetic characterization of rRNA gene sequences from DD-associated treponemes in the dairy herd environment. Although the spread of DD appears to be facilitated by wet floors covered with slurry, no DD-associated treponemes have been isolated from this environment previously. Consequently, there is a lack of knowledge about the spread of this disease among cows within a herd as well as between herds. To address the issue of DD infection reservoirs, we searched for evidence of DDassociated treponemes in fresh feces, in slurry, and in hoof lesions by deep sequencing of the V3 and V4 hypervariable regions of the 16S rRNA gene coupled with identification at the operational-taxonomic-unit level. Using treponeme-specific primers in this high-throughput approach, we identified small amounts of DNA (on average 0.6% of the total amount of sequence reads) from DD-associated treponemes in 43 of 64 samples from slurry and cow feces collected from six geographically dispersed dairy herds. Species belonging to the Treponema denticola/Treponema pedis-like and Treponema phagedenis-like phylogenetic clusters were among the most prevalent treponemes in both the dairy herd environment and the DD lesions. By the high-throughput approach presented here, we have demonstrated that cow feces and environmental slurry are possible reservoirs of DD-associated treponemes. This method should enable further clarification of the etiopathogenesis of DD. Bovine digital dermatitis (DD) was first described in 1974 and has since become a growing problem worldwide (1). The disease is characterized by focal proliferative to ulcerative dermatitis that is typically located on the plantar aspect of the hoof (1). The consequences of this disease are decreased animal welfare and serious economic losses for the farmers due to factors such as reduced milk yield and premature culling (2-4). DD is considered a multifactorial disease, and moist, unhygienic conditions, such as the slurry to which the animals are frequently exposed, are considered predisposing factors (5, 6). The bacteria most often associated with DD are spirochetes of the genus Treponema, which predominate in the deepest regions of DD lesions (7,8). To date, at least 20 different phylotypes have been identified from DD biopsy specimens; among these, Treponema phagedenis-like, Treponema medium/Treponema vincentii-like, Treponema denticolalike, and Treponema pedis phylotypes are highly associated with progression of the disease (9-12). DD also appears to be highly infectious based on the rapid intra-and interherd spread of the disease (6, 13, 14). The spread of DD in dairy cattle is facilitated by wet walking surfaces covered with feces (15), although the causative agents have not yet been found in slurry (16). Because the Treponema ...

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Targeting the Treponemal Microbiome of Digital Dermatitis Infections by High-Resolution Phylogenetic Analyses and Comparison with Fluorescent In Situ Hybridization

Cited by 59 publications

References 35 publications

Isolation of Digital Dermatitis Treponemes from Hoof Lesions in Wild North American Elk (Cervus elaphus) in Washington State, USA

Isolation of Digital Dermatitis Treponemes from Hoof Lesions in Wild North American Elk (Cervus elaphus) in Washington State, USA

Deep Sequencing Analysis Reveals Temporal Microbiota Changes Associated with Development of Bovine Digital Dermatitis

Discovery of Bovine Digital Dermatitis-Associated Treponema spp. in the Dairy Herd Environment by a Targeted Deep-Sequencing Approach

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