Differentiation of intestinal spirochaetes by multilocus enzyme electrophoresis analysis and 16S rRNA sequence comparisons

Stanton, Thad B.; Trott, Darren J.; Lee, Jae I.; McLaren, Andrew; Hampson, D.J.; Paster, B. J.; Jensen, Neil S.

doi:10.1111/j.1574-6968.1996.tb08046.x

Cited by 50 publications

(59 citation statements)

References 26 publications

(12 reference statements)

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“…Between species, the levels of DNA relatedness to heterologous reference strains ranged from 21 to 68%, with ATm values of more than 5°C. The DNA homology results indicate that rhea spirochete strain R-1 is an S. hyodysenteriae strain and that human spirochete strain WES-B is a strain of S. pilosicoli, which is consistent with the identification of these strains based on MEE analysis (35).…”

Section: Resultssupporting

confidence: 71%

“…The gas yields were calculated as micromoles per milliliter and do not reflect the actual concentrations of gases in the medium. quences of these two species are very similar (26,35). In addition, Brachyspira aalborgi could be differentiated from the Serpulina species since its amplicon was cleaved by MboII into two fragments that were 93 and 465 bp long (data not shown) and the amplicons from the Serpulina species were not cut by this enzyme.…”

Section: Resultsmentioning

confidence: 75%

“…For these new species we propose the names S. intermedia (previously called "S. intermedius") and S. murdochii (previously also called group B spirochetes) (19,20). Indeed, DNA relatedness results indicate that the groups of intestinal spirochete strains defined previously on the basis of MEE results (20,35) represent separate Serpulina species. Groups I, 111, and VI correspond to S. hyodysenteriae, S. innocens, and S. pilosicoli, respectively, while group I1 corresponds to S. intermedia sp.…”

Section: Discussionmentioning

confidence: 95%

“…intermedia strains have been classified by MEE analysis (MEE group I1 [35]) and by biochemical tests (group I1 spirochetes [7,81). This spirochete has DNA homology levels of 57, 45,28, and 37% with S. hyodysenteriae B78T, S. innocens B256T, S. pilosicoli P43/6/7gT, and S. murdochii 56-150T, respectively.…”

Section: Discussionmentioning

confidence: 99%

“…More recently, MEE analysis established two additional spirochete groups ( 3 9 , and comparisons of 16s ribosomal DNA (rDNA) sequences of intestinal spirochetes belonging to the seven MEE groups demonstrated that they are closely related to one another (26,35). The highly conserved 16s rDNA sequences of these intestinal spirochetes led to a recommendation that alternative techniques, such as DNA-DNA relative reassociation, should be used to define Serpulina species…”

Section: Swinementioning

confidence: 99%

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Recognition of Two New Species of Intestinal Spirochetes: Serpulina intermedia sp. nov. and Serpulina murdochii sp. nov.

Stanton

Fournié-Amazouz

Postić

et al. 1997

International Journal of Systematic Bacteriology

112

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On the basis of DNA-DNA hybridization data, nine intestinal spirochete strains were grouped into five genospecies. Three of these genospecies were previously recognized Serpulina species, Serpulina hyodysenteriae (type strain, B78), Serpulina innocens (type strain, B256), and Serpulina pilosicoli (type strain, P43/6/78; previously "Anguillina coli"). The other two genospecies were found to be new Serpulina species, for which we propose the names Serpulina intermedia sp. nov. (with type strain PWS/A) and Serpulina murdochii sp. nov. (with type strain 56-150). S. intermedia and S. murdochii cells had a typical spirochete ultrastructure with 22 to 28 periplasmic flagella per cell. Various soluble sugars were growth substrates for S. intermedia and S. murdochii. During growth in basal heart infusion broth supplemented with fetal calf serum beneath an 0,-N, (1:99) atmosphere, cells of these new species consumed oxygen and glucose and produced H,, CO,, acetate, butyrate, and ethanol. The G+C content of the DNA of S. murdochii 56-150T was 27 mol%, and the G+C content of the DNA of S. intermedia PWS/AT was 25 mol%. In addition, a restriction fragment length polymorphism-PCR assay for the detection of intestinal spirochetes was developed. The assay was based on generation and restriction endonuclease analysis (with HinfI, TaqI, Sau3A, and 4421011) of a 558-bp amplicon of ribosomal DNA (rDNA) encoding 16s rRNA. The PCR amplification was specific for Serpulina species and Brachyspira aalborgi. Four restriction digest patterns were found for the five Serpulina species. HinfI restriction differentiated S. murdochii and S. innocens from the other species. Sau3A and TaqI restrictions gave unique fragment patterns for S. murdochii and S. pilosicoli, respectively. S. hyodysenteriae and S. intermedia DNAs gave the same fragment pattern regardless of the enzyme tested. B. aalborgi was differentiated from the Serpulina species by MboII digestion of the 16s rDNA amplicon. Swine (367 40) dysentery (13) and porcine intestinal spirochetosis can be differentiated by their clinical signs. Swine dysentery is a severe, mucohemokhagic diarrheal disease that sometimes leads to death. Porcine intestinal spirochetosis is a mucus-containing, nonbloody diarrheal disease which leads to a poor growth rate in young pigs. The etiological agents of these two diseases are separate species of intestinal spirochetes belonging to the genus Serpulina, the strongly hemolytic organism Selpulina hyodysenteriae (13) and the weakly hemolytic organism Serpulina pilosicoli (formerly called "Anguillina coli") (40), respectively. Some human intestinal spirochetes (1, 18, 29) have been identified as S. pilosicoli (25, 41). Other previously characterized spirochetes found in the gastrointestinal tracts and feces of animals and humans include two nonpathogenic species from swine, Selpulirza innocens (31) and Treponema succinifaciens (5), and a human spirochete, Brachyspira aalborgi, which is believed to be a commensal (14). There undoubtedly are other intestinal spir...

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Section: Resultssupporting

confidence: 71%

Section: Resultsmentioning

confidence: 75%