Monitoring experimental swine dysentery: rectal swab blood test and Serpulina (Treponema) hyodysenteriae detection

Stanton, Thad B.; Jensen, Neil S.

doi:10.1016/0378-1135(93)90064-e

Cited by 5 publications

(4 citation statements)

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“…Blood in feces was assessed visually or by an occult blood test (44). Shedding of B. hyodysenteriae cells in feces was monitored by direct microscopy and by culturing rectal swab samples on TSB agar plates containing spectinomycin (39).…”

Section: Methodsmentioning

confidence: 99%

Isolation, Oxygen Sensitivity, and Virulence of NADH Oxidase Mutants of the Anaerobic SpirocheteBrachyspira(Serpulina)hyodysenteriae, Etiologic Agent of Swine Dysentery

Stanton

Rosey²,

Kennedy³

et al. 1999

Appl Environ Microbiol

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Brachyspira (Serpulina)hyodysenteriae, the etiologic agent of swine dysentery, uses the enzyme NADH oxidase to consume oxygen. To investigate possible roles for NADH oxidase in the growth and virulence of this anaerobic spirochete, mutant strains deficient in oxidase activity were isolated and characterized. The cloned NADH oxidase gene (nox; GenBank accession no. U19610 ) on plasmid pER218 was inactivated by replacing 321 bp of coding sequence with either a gene for chloramphenicol resistance (cat) or a gene for kanamycin resistance (kan). The resulting plasmids, respectively, pCmΔNOX and pKmΔNOX, were used to transform wild-type B. hyodysenteriae B204 cells and generate the antibiotic-resistant strains Nox-Cm and Nox-Km. PCR and Southern hybridization analyses indicated that the chromosomal wild-type nox genes in these strains had been replaced, through allelic exchange, by the inactivatednox gene containing cat or kan. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblot analysis revealed that both nox mutant cell lysates were missing the 48-kDa Nox protein. Soluble NADH oxidase activity levels in cell lysates of Nox-Cm and Nox-Km were reduced 92 to 96% compared to the activity level in parent strain B204. In an aerotolerance test, cells of both nox mutants were at least 100-fold more sensitive to oxygen exposure than were cells of the wild-type parent strain B204. In swine experimental infections, bothnox mutants were less virulent than strain B204 in that fewer animals were colonized by the mutant cells and infected animals displayed mild, transient signs of disease, with no deaths. These results provide evidence that NADH oxidase serves to protect B. hyodysenteriae cells against oxygen toxicity and that the enzyme, in that role, contributes to the pathogenic ability of the spirochete.

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Section: Methodsmentioning

confidence: 99%

Isolation, Oxygen Sensitivity, and Virulence of NADH Oxidase Mutants of the Anaerobic SpirocheteBrachyspira(Serpulina)hyodysenteriae, Etiologic Agent of Swine Dysentery

Stanton

Rosey²,

Kennedy³

et al. 1999

Appl Environ Microbiol

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“…In 85.7% of B. hyodysenteriae culture positive samples the direct stains did not reveal the presence of spirochaete in three samples (14.3%). Stanton and Jensen (1993) also found that 12.5% of samples positive by selective culture were negative by direct microscopy in experimental swine dysentery in postweaning piglets. This result is not unexpected since culture methods using TSA with blood and antimicrobial agents are more sensitive than direct microscopy for detection of B. hyodysenteriae (Stanton and Jensen, 1993).…”

Section: Resultsmentioning

confidence: 88%

“…Stanton and Jensen (1993) also found that 12.5% of samples positive by selective culture were negative by direct microscopy in experimental swine dysentery in postweaning piglets. This result is not unexpected since culture methods using TSA with blood and antimicrobial agents are more sensitive than direct microscopy for detection of B. hyodysenteriae (Stanton and Jensen, 1993). But interestingly, four out of 25 samples (16%) (one sample of faeces, three samples of colonic mucosa) positive by direct microscopy were negative for B. hyodysenteriae, but one sample was positive for B. intermedia by selective culture.…”

Section: Resultsmentioning

confidence: 88%

“…But interestingly, four out of 25 samples (16%) (one sample of faeces, three samples of colonic mucosa) positive by direct microscopy were negative for B. hyodysenteriae, but one sample was positive for B. intermedia by selective culture. Olson and Fales (1983) and Stanton and Jensen (1993) similarly reported that 24-35% of rectal swabs contained visible spirochaetes but did not give any positive cultures. These authors suggested that the optimal methods for detecting B. hyodysenteriae in faeces should include both direct microscopy and selective culture.…”

Section: Resultsmentioning

confidence: 94%

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Brachyspira hyodysenteriae: detection, identification and antibiotic susceptibility

Novotná¹,

Skardová²

2002

Vet. Med. - Czech

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101 samples of faeces, colonic mucosa and rectal swabs taken from 100 pigs (29 commercial herds) were cultivated on Trypticase Soy Agar with 5% of sheep blood, spectinomycin (200 mg/l), vancomycin (50 mg/l), rifampicin (12.5 mg/l) and colistin (12.5 mg/l). Plates were incubated in an anaerobic container at 37°C for 5–7 days. 25 samples (10 faeces, 15 scrapings of colonic mucosa) were examined by darkfield microscopy for the presence of spirochaetes. In 80 samples (21 faeces, 31 rectal swabs, 28 scrapings of colonic mucosa) from diarrhoeic pigs 44 isolates of spirochaetes were identified by PCR method as Brachyspira hyodysenteriae. In 21 samples (20 rectal swabs, 1 scraping of colonic mucosa) taken from apparently healthy pigs in herds with swine dysentery were isolated weakly haemolytic spirochaetes: B. intermedia in 5 samples and phenotypic group III brachyspirae in 4 samples. Minimal inhibitory concentration (MIC) of tylosin, tiamulin and valnemulin was determined by the agar dilution method. To tylosin 16 out of 17 strains of B. hyodysenteriae were resistant (MIC 64 and 256 µg/l), one strain was susceptible (MIC l µg/ml). To tiamulin 17 out of 19 strains were susceptible (MIC from 0.016 to 0.25 µg/ml), one strain was intermediately susceptible (MIC 2 µg/ml) and one resistant (MIC 32 µg/ml). To valnemulin 17 out of 19 strains were susceptible (MIC from 0.016 to 0.064 µg/ml), one strain was intermediately susceptible (MIC2µg/ml) and one was resistant (MIC 8 µg/ml). Valnemulin resistant strain was also resistant to tiamulin.

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The Genus Brachyspira

Stanton¹

2006

The Prokaryotes

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Monitoring experimental swine dysentery: rectal swab blood test and Serpulina (Treponema) hyodysenteriae detection

Cited by 5 publications

References 8 publications

Isolation, Oxygen Sensitivity, and Virulence of NADH Oxidase Mutants of the Anaerobic SpirocheteBrachyspira(Serpulina)hyodysenteriae, Etiologic Agent of Swine Dysentery

Isolation, Oxygen Sensitivity, and Virulence of NADH Oxidase Mutants of the Anaerobic SpirocheteBrachyspira(Serpulina)hyodysenteriae, Etiologic Agent of Swine Dysentery

Brachyspira hyodysenteriae: detection, identification and antibiotic susceptibility

The Genus Brachyspira

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