Physical and genetic map of the Serpulina hyodysenteriae B78T chromosome

Zuerner, Richard L.; Stanton, Thad B.

doi:10.1128/jb.176.4.1087-1092.1994

Cited by 50 publications

(40 citation statements)

References 39 publications

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“…These markers are located on the opposite side of the S. hyodysenteriae chromosome (35), so there does not appear to be preferences in regards to chromosomal location for genetic transduction. Since VSH-1 does not form plaques, the transduction frequency estimates were based on the number of phage particles estimated from direct electron microscope counts.…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of VSH-1, a generalized transducing bacteriophage of Serpulina hyodysenteriae

et al. 1997

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Serpulina hyodysenteriae B204 cells treated with mitomycin (20 g of mitomycin/ml of culture broth) lysed and released bacteriophages. Bacteriophage particles, precipitated by using polyethylene glycol and purified by CsCl density gradient ultracentrifugation, had a buoyant density of 1.375 g/cm 3 and consisted of a head (45-nm diameter) and an ultrastructurally simple (noncontractile) tail (64 by 9 nm) composed of at least 13 proteins with molecular masses ranging between 13 and 101 kDa. The purified bacteriophage has been designated VSH-1 (VSH for virus of S. hyodysenteriae). VSH-1 was incapable of lytic growth on any of five intestinal spirochete strains, representing three Serpulina species. VSH-1 nucleic acid was determined to be approximately 7.5 kb in size and to be linear, double-stranded DNA based on differential staining with acridine orange, DNase I sensitivity, electrophoretic mobility, and contour length as measured by electron microscopy. Phage DNA digested by the restriction enzymes SspI, AseI, EcoRV, and AflII gave electrophoretic banding patterns nearly identical to those of digested chromosomal DNA from S. hyodysenteriae. Additionally, VSH-1 DNA fragments hybridized with probes complementary to S. hyodysenteriae chromosomal genes nox and flaA1. When purified bacteriophages induced from cultures of S. hyodysenteriae A203 (⌬flaA1 593-762::cat) were added to growing cells of strain A216 (⌬nox 438-760::kan), transductants (Cm r Km r ) were obtained at a frequency of 1.5 ؋ 10 ؊6 per phage particle (enumerated by electron microscopy). These findings indicate that induced VSH-1 virions package DNA of S. hyodysenteriae and are capable of transferring host genes between cells of that spirochete. To our knowledge, this is the first report of genetic transduction of a spirochete.Natural gene transfer mechanisms (conjugative plasmids and transducing bacteriophages) have not been demonstrated for spirochetes. Indirect evidence of lateral gene transfer has been obtained for Borrelia spp. (16). The existence of DNAfilled membrane vesicles on the surfaces of Borrelia burgdorferi cells has been reported, although their possible role in gene transfer is unknown (7). In addition, several extrachromosomal elements have been identified in B. burgdorferi, and an extrachromosomal plasmid of 2.6 kb has been isolated from Treponema denticola and characterized (10,25). In cultures of various spirochetes, bacteriophages have been observed free, attached to cells, or within cells as noted previously (9). Three lytic phages of Leptospira biflexa have been isolated and characterized (24). However, to our knowledge, there is no evidence that these three phages play a role in gene transfer.The spirochete Serpulina hyodysenteriae causes swine dysentery, an enteric disease producing a severe mucohemorrhagic diarrhea in infected swine (8). Potential genetic transfer elements for S. hyodysenteriae have been identified. Ritchie and colleagues observed bacteriophages with the same morphology in 18 different cultures of S. hyodysente...

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Section: Discussionmentioning

confidence: 99%

Purification and characterization of VSH-1, a generalized transducing bacteriophage of Serpulina hyodysenteriae

et al. 1997

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“…PDD spirochetes were grown to an OD 620 of 0.8 in 100 ml of OTIS broth and encapsulated in agarose beads, and genomic DNA was prepared as described previously (47). Encapsulated DNA was digested at 37°C with approximately 10 U of restriction endonuclease for 4 to 6 h, then separated in 1% agarose gels by clamped homogenous electric field electrophoresis as described previously (47).…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2003

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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)...

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“…Five isolates including Brachyspira aalborgi (Hovind-Hougen et al, 1982) yielded too few fragments with SmaI and SacII to be used in the statistical analyses and were therefore removed from the study. The size of the whole intestinal spirochaete genome was estimated to be approximately 2.6-3 M b using PFGE following digestion with Not1 which has a single restriction site (Zuerner & Stanton, 1994). For each isolate the approximate genome size was obtained by adding together the sizes of the resolved restriction fragments and was found to be 3 Mb.…”

Section: Quantitative Evaluation Of Macrorestriction Fragment Profilesmentioning

confidence: 99%

Sub-specific differentiation of intestinal spirochaete isolates by macrorestriction fragment profiling

1997

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Macrorestriction fragment profile analysis by PFGE was used to distinguish intestinal spirochaetes, some of which were isolated from cases of swine dysentery and intestinal spirochaetosis in humans, pigs, mice, chickens and dogs. Macrorestriction fragment profiles using SmaI and Sacll restriction enzymes were produced and used in statistical analysis. This permitted the division of the isolates into two major clusters. One cluster contained isolates which were identified as Sepulina pilosicoli and the second cluster contained isolates identified as Serpuline hyodysenteriae by immunoblotting with species-specific mAbs. Both species contained sub-specif ic groups, although these rarely correlated with the source of the isolates. We conclude that PFGE is capable of sub-specif ic differentiation of intestinal spirochaetes, but that the current species contain a large variety of genotypes among which crossspecies transmission may be feasible.

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Physical and genetic map of the Serpulina hyodysenteriae B78T chromosome

Cited by 50 publications

References 39 publications

Purification and characterization of VSH-1, a generalized transducing bacteriophage of Serpulina hyodysenteriae

Purification and characterization of VSH-1, a generalized transducing bacteriophage of Serpulina hyodysenteriae

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

Sub-specific differentiation of intestinal spirochaete isolates by macrorestriction fragment profiling

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