The phylogeny of intestinal porcine spirochetes (Serpulina species) based on sequence analysis of the 16S rRNA gene

Pettersson, Bertil; Fellström, Claes; Andersson, Anders F.; Uhlén, Mathias; Gunnarsson, A.; Johansson, Karl‐Erik

doi:10.1128/jb.178.14.4189-4199.1996

Cited by 54 publications

(69 citation statements)

References 57 publications

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“…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: 80%

“…Thus, besides the strongly hemolytic organism S. hyodysenteriae, four weakly hemolytic species are found in the genus Serpulina. These species roughly correlate with the biochemical-16s rRNA groups devised by Fellstrom et al (7,8,26), with S. intermedia representing their group I1 spirochetes, S. innocens representing groups IIIb and IIIc, S. murdochii representing group IIIa, and S. pilosicoli representing group IV. S. murdochii and S. intermedia cells are similar in ultrastructure to cells of S. hyodysenteriae.…”

Section: Discussionmentioning

confidence: 84%

“…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: Resultsmentioning

confidence: 80%

Section: Discussionmentioning

confidence: 84%

Section: Swinementioning

confidence: 99%

See 1 more Smart Citation

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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“…In the case of B. pilosicoli, the various isolates and the two B. pilosicoli sequences amplified from PET were closely clustered near their type strain, P43/6/78 T . The relatively large distance between B. aalborgi and these other six Brachyspira species, compared with the relatively small distances between the latter species, has been reported in a number of other studies (Lee & Hampson, 1994;Pettersson et al, 1996Pettersson et al, , 2000Stanton et al, 1996). At the time that it was proposed that the former Serpulina species be included in the genus Brachyspira (Ochiai et al, 1997), this relatively large 16S rDNA sequence difference between the Brachyspira and Serpulina lineages caused disquiet about the proposal (Hampson, 2000).…”

Section: Discussionmentioning

confidence: 99%

Analysis of genetic variation in Brachyspira aalborgi and related spirochaetes determined by partial sequencing of the 16S rRNA and NADH oxidase genes

Mikosza

Munshi

Hampson

2004

Journal of Medical Microbiology

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The purpose of this study was to investigate genetic variation in the anaerobic intestinal spirochaete Brachyspira aalborgi by partial sequencing of the 16S rRNA and NADH oxidase genes. The spirochaete is poorly cultivable; hence, only six isolates were available for analysis. Additional sequences were amplified from DNA extracted from fixed colorectal biopsies from 26 patients with histological evidence of intestinal spirochaetosis, and from the faeces of six non-human primates (NHP). Multiple biopsies from sites along the large intestine were tested from three of the 26 patients. Sequences from two biopsies were closely related to those of the spirochaete Brachyspira pilosicoli. Eight B. aalborgi-like 16S rDNA sequences were generated from the biopsies from the other 24 patients, and four from the NHP faeces. The B. aalborgi 16S rDNA sequences were divided into three clusters, 1, 2 and 4, with individual sequence similarities to the type strain ranging from 97 . 49 to 100 %. All human isolates of B. aalborgi were located in cluster 1, as was the sequence of the so-called 'Brachyspira ibaraki'. All four 16S rDNA sequences from the NHP faeces and the two NHP isolates of B. aalborgi were located in cluster 4, which was distinct. Cluster 4 may represent a novel Brachyspira species. Evidence for multiple strains of B. aalborgi or other Brachyspira species was found in biopsies from two patients. In the three individuals from whom multiple biopsies were amplified, the sequences at each intestinal site were the same, indicating the presence of one dominant strain.

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“…16S rDNA sequences of 11 of the selected Brachyspira isolates from farmed and wild-living mallards were determined as described by Pettersson et al (1996), with some modifications. The 16S rRNA gene was first amplified with the PCR primers listed in Table 2.…”

Section: Methodsmentioning

confidence: 99%

Brachyspira hyodysenteriae and other strongly -haemolytic and indole-positive spirochaetes isolated from mallards (Anas platyrhynchos)

Jansson¹,

Johansson²,

Olofsson³

et al. 2004

Journal of Medical Microbiology

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The aims of the current study were to collect intestinal spirochaetes (genus Brachyspira) from farmed and wild mallards (Anas platyrhynchos) and to identify and classify those isolates that phenotypically resembled Brachyspira hyodysenteriae, an enteric pathogen of pigs. The isolation rate of Brachyspira spp. was high from both farmed (93 %) and wild mallards (78 %). In wild mallards, it appeared that Brachyspira spp. were more likely to be found in migratory birds (multivariate analysis: RR 1 . 8, 95 % CI 1 . 1-3 . 1) than in mallards sampled in a public park. Pure cultures of putative B. hyodysenteriae were obtained from 22 birds. All five isolates from farmed mallards and ten randomly selected isolates with this phenotype were used for further studies. All isolates from farmed mallards and two of the isolates from wild mallards were PCR-positive for the tlyA gene of B. hyodysenteriae. Two isolates from farmed mallards were selected for pulsed field gel electrophoresis (PFGE) and randomly amplified polymorphic DNA (RAPD) analysis. These isolates clustered with the type and reference strains of B. hyodysenteriae. 16S rDNA sequence analysis performed on 11 of the strains showed that they were all closely related to each other and to the B. hyodysenteriae-Brachyspira intermedia cluster. Three of the mallard isolates had 16S rDNA sequences that were identical to those of B. hyodysenteriae strains R1 and NIV-1 previously isolated from common rheas (Rhea americana). To conclude, the isolates from farmed mallards and two isolates from wild mallards were classified as B. hyodysenteriae based on the fact that they could not be differentiated by any of the applied methods from type, reference and field strains of B. hyodysenteriae. The remaining isolates could not be assigned irrefutably to any of the presently recognized Brachyspira species. These results point to a broader host spectrum of B. hyodysenteriae than is generally recognized, and to the presence in mallards of strongly â-haemolytic and indole-producing spirochaetes that possess many, but not all, of the currently recognized characteristics of B. hyodysenteriae.

show abstract

The phylogeny of intestinal porcine spirochetes (Serpulina species) based on sequence analysis of the 16S rRNA gene

Cited by 54 publications

References 57 publications

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

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

Analysis of genetic variation in Brachyspira aalborgi and related spirochaetes determined by partial sequencing of the 16S rRNA and NADH oxidase genes

Brachyspira hyodysenteriae and other strongly -haemolytic and indole-positive spirochaetes isolated from mallards (Anas platyrhynchos)

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