Biochemical and molecular characterization of Treponema phagedenis-like spirochetes isolated from a bovine digital dermatitis lesion

Abstract

BackgroundBovine papillomatous digital dermatitis (DD) is the leading cause of lameness in dairy cattle and represents a serious welfare and economic burden. Found primarily in high production dairy cattle worldwide, DD is characterized by the development of an often painful red, raw ulcerative or papillomatous lesion frequently located near the interdigital cleft and above the bulbs of the heel. While the exact etiology is unknown, several spirochete species have been isolated from lesion material. Four isolates of Treponema phagedenis-like spirochetes were isolated from dairy cows in Iowa. Given the distinct differences in host, environmental niche, and disease association, a closer analysis of phenotypic characteristics, growth characteristics, and genomic sequences of T. phagedenis, a human genitalia commensal, and the Iowa DD isolates was undertaken.ResultsPhenotypically, these isolates range from 8.0 to 9.7 μm in length with 6–8 flagella on each end. These isolates, like T. phagedenis, are strictly anaerobic, require serum and volatile fatty acids for growth, and are capable of fermenting fructose, mannitol, pectin, mannose, ribose, maltose, and glucose. Major glucose fermentation products produced are formate, acetate, and butyrate. Further study was conducted with a single isolate, 4A, showing an optimal growth pH of 7.0 (range of 6–8.5) and an optimal growth temperature of 40°C (range of 29°C-43°C). Comparison of partial genomic contigs of isolate 4A and contigs of T. phagedenis F0421 revealed > 95% amino acid sequence identity with amino acid sequence of 4A. In silico DNA-DNA whole genome hybridization and BLAT analysis indicated a DDH estimate of >80% between isolate 4A and T. phagedenis F0421, and estimates of 52.5% or less when compared to the fully sequenced genomes of other treponeme species.ConclusionUsing both physiological, biochemical and genomic analysis, there is a lack of evidence for difference between T. phagedenis and isolate 4A. The description of Treponema phagedenis should be expanded from human genital skin commensal to include being an inhabitant within DD lesions in cattle.

Isolation and identification of Treponema pedis and Treponema phagedenis-like organisms from bovine digital dermatitis lesions found in dairy cattle in Turkey

“…The enzyme pattern of reactivity of current isolates with positive results for esterase C4, esterase lipase C8, leucine arylamidase, trypsin, and chymotrypsin (group I isolates) was similar to UK bovine treponemes of group 3 and T. pedis except for leucine arylamidase (Evans et al, 2008 and2009a, respectively). Group II isolates shared similar reactivity with T. phagedenis Kazan except for β-galactosidase (Wilson-Welder et al, 2013). The enzyme profiles of the current isolates produced a specific pattern that differed from the genus Treponema shown in Table 4.…”

“…Although Holstein or Friesian-Holstein cows form the backbone of the dairy industry worldwide, managed usually in larger herds and housed partially or permanently throughout the year, Simmen- Table 2. Bacterial size and flagella number of treponemes identified in this study, compared with Treponema pedis and Treponema phagedenis characteristics described by Evans et al (2008) and Wilson-Welder et al (2013) Treponeme and numbers examined (n) (Evans et al, 2008) 3:6:3 5:10:5 5 16 0.2 0.3 Reference for T. phagedenis (Wilson-Welder et al, 2013) 7:14:7 9:18:9 8 9.7 0.3 0.35 Figure 3. Neighbor-joining phylogenetic tree with respect to 16S rRNA gene sequence comparisons of ~1,400 aligned bases (TOVAG112O888 in this study).…”

“…Histopathological examination of DD lesion biopsies have shown consistently that spirochete-like microorganisms are present (Demirkan et al, 1999;Santos et al, 2012), and these are now known to be treponemes that appear phylogenetically and serologically diverse within lesions and in different countries (Schrank et al, 1999;Stamm et al, 2002;Elliott et al, 2007; , 2009b;Yano et al, 2010;Wilson-Welder et al, 2013;Gomez et al, 2014;Nascimento et al, 2015). These treponemes are clustered within 3 groups, described as Treponema medium/Treponema vincentii-like, Treponema phagedenis-like, and Treponema denticola/Treponema putidum-like (Evans et al, 2008), the latter now classified as a new species, Treponema pedis (Evans et al, 2009a).…”

Conservation of the Host-Interacting Proteins Tp0750 and Pallilysin among Treponemes and Restriction of Proteolytic Capacity to Treponema pallidum

“…Studies have shown that T. phagedenis migrates intercellularly deep within the epidermis and, if left untreated, digital dermatitis lesions eventually affect the whole hoof (52,53). Recent biochemical, molecular, and physiological characterizations, together with genome-wide comparisons of T. phagedenis strain F0421 and bovine T. phagedenis isolates (including strain 4A), indicated that the two spirochetes are almost identical in terms of biochemistry and genetics, leading to the conclusion that these bacteria belong to the same species and that the current description of T. phagedenis should be broadened to include both commensal strains of humans and pathogenic strains in cattle (54).…”

Treponemes Detected in Digital Dermatitis Lesions in Brazilian Dairy Cattle and Possible Host Reservoirs of Infection

“…The ruminal environment may favor the survival of certain treponemes, because an environment rich in volatile fatty acids favors T. phagedenis (21). Further, the ability of treponemes to colonize different environments depends, among other factors, on the presence of periplasmic flagella for better propulsion through highly viscous or viscoelastic material (22), such as ruminal fluid and humid claw skin.…”