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

Abstract

The isolation and identification of microorganisms associated with digital dermatitis (DD) in Turkish dairy cattle was investigated using punch skin biopsy samples from typical clinical lesions; they were collected from dairy farms and abattoirs in 5 different geographical locations in Turkey. Morphological characteristics and flagellation types were examined using a transmission electron microscope, and their enzyme profiles by enzyme activation kits; their catalase reaction characteristics were evaluated by the addition of 3% HO. Their phylogenies were identified using 16S rRNA and the results compared with known gene bank data. Bacterial cells were 5.0 to 18.2 µm long, 0.2 to 0.5 µm wide, and their minimum number of periplasmic flagellum was 4 (4:8:4) with a maximum of 8 (8:16:8). All isolates were catalase negative. Of the spirochetes isolated and identified, group I organisms showed close similarity with Treponema pedis (99% genetic homology), whereas those in group II were similar to Treponema phagedenis (98% homology). This is the first report of specific sub-groups of Treponema spp., isolated from Turkish dairy cattle presenting with DD lesions, being associated with this disease; these morphotypes were similar to those found globally in housed dairy cattle units and are probably significant microorganisms associated with the aetiopathogensis of this infectious disease causing acute bovine lameness. These results suggest that the distribution of DD-associated treponemes is not specific to particular geographic regions of Turkey.

Genotypic and Phenotypic Characterization of Treponema phagedenis from Bovine Digital Dermatitis

“…Difficulties in culturing BDD-associated treponemes, due to the fact of their fastidiousness has restricted the number of isolates and the recovery of treponemes collected from lesion samples. In Treponema -associated studies, concerns regarding the difficulty of isolating a pure culture has been emphasized [ 14 , 24 ]. Extensive cleaning was crucial in the elimination of contaminants, and the increased antibiotic concentrations allowed treponemal growth while inhibiting others’ growth.…”

“…Treponema denticola displayed an iron acquisition mechanism that involved binding a 47 kDa outer membrane sheath with hemin, an important iron source derived from hemoglobin [ 28 ]. Varying degrees of hemolysis by T. phagedenis from BDD have been reported [ 1 , 2 , 14 , 16 ], but this property still requires further investigation since BDD often involves a bleeding response. Furthermore, round bodies have been observed in other Spirochetes , especially on human pathogens, Borrelia burgdorferi, and Treponema pallidum subsp.…”

“… * Isolates in this study, a [ 18 ]; b [ 16 ]; c [ 15 ]; d [ 2 ]; e [ 14 ]; f [ 1 ]. (1) alkaline phosphatase, (2) C4 esterase, (3) C8 esterase lipase, (4) C14 lipase, (5) leucine arylamidase, (6) valine arylamidase, (7) cystine arylamidase, (8) trypsin, (9) α-chymotrypsin, (10) acid phosphatase, (11) Naphtol-AS-BI-phosphohydrolase, (12) α-galactosidase, (13) β-galactosidase, (14) β-glucuronidase, (15) α-glucosidase, (16) β-glucosidase, (17) N-acetyl β-glucosaminidase, (18) α-mannosidase, (19) α-fucosidase.…”

“…The fastidiousness of Treponema makes it difficult to culture in vitro which has limited the success of studies aimed at determining the pathogenic mechanisms of these bacteria. Only a few researchers have successfully isolated and characterized BDD-associated treponemes [ 1 , 2 , 11 , 14 , 15 , 16 , 17 , 18 ]. Recently, an investigation on BDD in Korea investigated the microbiome diversity and the dominant pathogens in lesions [ 9 ].…”

Treponema spp. Isolated from Bovine Digital Dermatitis Display Different Pathogenicity in a Murine Abscess Model

“…are consistently identified in active bovine DD lesions [ 8 , 10 , 11 , 12 ], the exact role of these spirochetes in the aetiopathogenesis of DD remains unclear [ 13 ]. Treponema phagedenis , T. medium , T. pedis are among the most commonly detected species in DD lesions [ 8 , 10 , 11 , 14 , 15 ]. Interestingly, DD-associated treponemes have also been associated with other hoof-related animal diseases, including contagious ovine digital dermatitis [ 16 ] and elk hoof disease [ 17 ].…”