1887

Abstract

Porcine intestinal spirochaetes are fastidious anaerobic organisms and, as a consequence, it has been necessary to develop various protocols to enhance their isolation from or detection in faeces. Immunomagnetic separation (IMS) is a method developed recently to improve separation of target cells from mixed cell suspensions. The purpose of the present study was to compare the relative sensitivity of IMS for isolation of and with current routine diagnostic methods (culture on selective media and PCR) for detection of these micro-organisms in pig faeces. Neither direct nor indirect IMS methods enhanced the sensitivity of detection of either organism when performed with the recommended washings during sample processing. Performance of the IMS procedure without washing gave sensitivity at levels similar to direct culture onto selective medium. Further development of IMS techniques is required to improve isolation rates of species from faecal samples.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.05500-0
2004-04-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jmm/53/4/JM530407.html?itemId=/content/journal/jmm/10.1099/jmm.0.05500-0&mimeType=html&fmt=ahah

References

  1. Achacha M., Messier S. 1992; Comparison of six different culture media for isolation of Treponema hyodysenteriae . J Clin Microbiol 30:249–251
    [Google Scholar]
  2. Atyeo R. F., Oxberry S. L., Combs B. G., Hampson D. J. 1998; Development and evaluation of polymerase chain reaction tests as an aid to diagnosis of swine dysentery and intestinal spirochaetosis. Lett Appl Microbiol 26:126–130 [CrossRef]
    [Google Scholar]
  3. Barcellos D. E. S., de Uzeda M., Ikuta N., Lunge V. R., Fonseca A. S. K., Kader I. I. T., Duhamel G. E. 2000; Identification of porcine intestinal spirochaetes by PCR-restriction fragment length polymorphism analysis of ribosomal DNA encoding 23S rRNA. Vet Microbiol 75:189–198 [CrossRef]
    [Google Scholar]
  4. Calderaro A., Merialdi G., Perini S., Ragni P., Guegán R., Dettori G., Chezzi C. 2001; A novel method for the isolation of Brachyspira ( Serpulina ) hyodysenteriae from pigs with swine dysentery in Italy. Vet Microbiol 80:47–52 [CrossRef]
    [Google Scholar]
  5. Chapman P. A., Siddons C. A. 1996; A comparison of immunomagnetic separation and direct culture for the isolation of verocytotoxin-producing Escherichia coli O157 from cases of bloody diarrhoea, non-bloody diarrhoea and asymptomatic contacts. J Med Microbiol 44:267–271 [CrossRef]
    [Google Scholar]
  6. Cooper C., Cotton D. W., Hudson M. J., Kirkham N., Wilmott F. E. 1986; Rectal spirochaetosis in homosexual men: characterisation of the organism and pathophysiology. Genitourin Med 62:47–52
    [Google Scholar]
  7. Duhamel G. E., Muniappa N., Gardner I., Anderson M. A., Blanchard P. C., DeBey B. M., Mathiesen M. R., Walker R. L. 1995; Porcine colonic spirochetosis: a diarrhoeal disease associated with newly recognised species of intestinal spirochaetes. Pig J 35:101–110
    [Google Scholar]
  8. Duhamel G. E., Trott D. J., Muniappa N., Mathiesen M. R., Tarasiuk K., Lee J. I., Hampson D. J. 1998; Canine intestinal spirochetes consist of Serpulina pilosicoli and a newly identified group provisionally designated ‘‘ Serpulina canis ’’ sp.nov. J Clin Microbiol 36:2264–2270
    [Google Scholar]
  9. Dwars R. M., Davelaar F. G., Smith H. F. 1992; Infection of broiler chicks ( Gallus domesticus ) with human intestinal spirochaetes. Avian Pathol 21:559–568 [CrossRef]
    [Google Scholar]
  10. Enroth H., Engstrand L. 1995; Immunomagnetic separation and PCR for detection of Helicobacter pylori in water and stool specimens. J Clin Microbiol 33:2162–2165
    [Google Scholar]
  11. Fellström C., Gunnarsson A. 1995; Phenotypical characterisation of intestinal spirochaetes isolated from pigs. Res Vet Sci 59:1–4 [CrossRef]
    [Google Scholar]
  12. Fellström C., Pettersson B., Thomson J., Gunnarsson A., Persson M., Johansson K.-E. 1997; Identification of Serpulina species associated with porcine colitis by biochemical analysis and PCR. J Clin Microbiol 35:462–467
    [Google Scholar]
  13. Fellström C., Zimmerman U., Aspan A., Gunnarsson A. 2001; The use of culture, pooled samples and PCR for the identification of herds infected with Brachyspira hyodysenteriae . Anim Health Res Rev 2:37–43
    [Google Scholar]
  14. Heuvelink A. E., van den Biggelaar F. L. A. M., de Boer E., Herbes R. G., Melchers W. J. G., Huis in't Veld J. H. J., Monnens L. A. H. 1998; Isolation and characterization of verocytotoxin-producing Escherichia coli O157 strains from Dutch cattle and sheep. J Clin Microbiol 36:878–882
    [Google Scholar]
  15. Hommez J., Castryck F., Haesebrouck F., Devriese L. A. 1998; Identification of porcine Serpulina strains in routine diagnostic bacteriology. Vet Microbiol 62:163–169 [CrossRef]
    [Google Scholar]
  16. Jenkinson S. R., Wingar C. R. 1981; Selective medium for the isolation of Serpulina hyodysenteriae . Vet Rec 109:384–385 [CrossRef]
    [Google Scholar]
  17. Kunkle R. A., Kinyon J. M. 1988; Improved selective medium for the isolation of Treponema hyodysenteriae . J Clin Microbiol 26:2357–2360
    [Google Scholar]
  18. La T., Phillips N. D., Hampson D. J. 2003; Development of a duplex PCR assay for detection of Brachyspira hyodysenteriae and Brachyspira pilosicoli in pig feces. J Clin Microbiol 41:3372–3375 [CrossRef]
    [Google Scholar]
  19. Lee B. J., Hampson D. J. 1995; A monoclonal antibody reacting with the cell envelope of spirochaetes isolated from cases of intestinal spirochaetosis in pigs and humans. FEMS Microbiol Lett 131:179–184 [CrossRef]
    [Google Scholar]
  20. Lee B. J., Hampson D. J. 1996; Production and characterisation of a monoclonal antibody to Serpulina hyodysenteriae . FEMS Microbiol Lett 136:193–197 [CrossRef]
    [Google Scholar]
  21. Leser T. D., Møller K., Jensen T. K., Jorsal S. E. 1997; Specific detection of Serpulina hyodysenteriae and potentially pathogenic weakly β-haemolytic porcine intestinal spirochetes by polymerase chain reaction targeting 23S rDNA. Mol Cell Probes 11:363–372 [CrossRef]
    [Google Scholar]
  22. Lysons R. J., Lemcke R. M. 1983; Swine dysentery: to isolate or to fluoresce?. Vet Rec 112:203– 203
    [Google Scholar]
  23. Marshall K. C., Stout R., Mitchell R. 1971; Mechanism of the initial events in the sorption of marine bacteria to surfaces. J Gen Microbiol 68:337–348 [CrossRef]
    [Google Scholar]
  24. McLaren A. J., Trott D. J., Swayne D. E., Oxberry S. L., Hampson D. J. 1997; Genetic and phenotypic characterization of intestinal spirochetes colonizing chickens and allocation of known pathogenic isolates to three distinct genetic groups. J Clin Microbiol 35:412–417
    [Google Scholar]
  25. Mikosza A. S. J., La T., Margawani K. R., Brooke C. J., Hampson D. J. 2001; PCR detection of Brachyspira aalborgi and Brachyspira pilosicoli in human faeces. FEMS Microbiol Lett 197:167–170 [CrossRef]
    [Google Scholar]
  26. Park N. Y., Chung C. Y., McLaren A. J., Atyeo R. F., Hampson D. J. 1995; Polymerase chain reaction for identification of porcine spirochaetes recovered from cases of intestinal spirochaetosis. FEMS Microbiol Lett 125:225–229 [CrossRef]
    [Google Scholar]
  27. Rasmussen A. M., Smeland E., Erikstein B. K., Caignault L., Funderud S. 1990; A new method for detachment of Dynabeads from positively selected B lymphocytes. J Immunol Methods 146:195–202
    [Google Scholar]
  28. Sambrook J., Fritch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  29. Songer J. G., Kinyon J. M., Harris D. L. 1976; Selective medium for isolation of Treponema hyodysenteriae . J Clin Microbiol 4:57–60
    [Google Scholar]
  30. Stanton T. B., Postic D., Jensen N. S. 1998; Serpulina alvinipulli sp.nov., a new Serpulina species that is enteropathogenic for chickens. Int J Syst Bacteriol 48:669–676 [CrossRef]
    [Google Scholar]
  31. Stanton T. B., Rosey E. L., Kennedy M. J., Jensen N. S., Bosworth B. T. 1999; Isolation, oxygen sensitivity, and virulence of NADH oxidase mutants of the anaerobic spirochaete Brachyspira ( Serpulina ) hyodysenteriae, etiologic agent of swine dysentery. Appl Environ Microbiol 65:5028–5034
    [Google Scholar]
  32. Stone G. G., Oberst R. D., Hays M. P., McVey S., Chengappa M. M. 1994; Detection of Salmonella serovars from clinical samples by enrichment broth cultivation-PCR procedure. J Clin Microbiol 32:1742–1749
    [Google Scholar]
  33. Surawicz C. M., Roberts P. L., Rompalo A., Quinn T. C., Holmes K. K., Stamm W. E. 1987; Intestinal spirochetosis in homosexual men. Am J Med 82:587–592 [CrossRef]
    [Google Scholar]
  34. Takeuchi A., Jervis H. R., Nakazawa H., Robinson D. M. 1974; Spiral-shaped organisms on the surface colonic epithelium of the monkey and man. Am J Clin Nutr 27:1287–1296
    [Google Scholar]
  35. Taylor D. J., Alexander T. J. L. 1971; The production of dysentery in swine by feeding cultures containing a spirochaete. Br Vet J 127:58–61
    [Google Scholar]
  36. Taylor D. J., Simmons J. R., Laird H. M. 1980; Production of diarrhoea and dysentery in pigs by feeding pure cultures differing from Treponema hyodysenteriae . Vet Rec 106:326–332 [CrossRef]
    [Google Scholar]
  37. Taylor M. J., Ellis W. A., Montgomery J. M., Yan K. T., McDowell S. W. J., Mackie D. P. 1997; Magnetic immunocapture PCR assay (MIPA): detection of Leptospira borgpetersenii serovar hardjo . Vet Microbiol 56:135–145 [CrossRef]
    [Google Scholar]
  38. Teran-Dianderas M. 1997; Rapid detection and identification of porcine Serpulina sp. by polymerase chain reaction (PCR) and restriction fragment analysis. MSc thesis University of Edinburgh; Scotland, UK:
  39. Vanrobaeys M., de Herdt P., Ducatelle R., Devriese L. A., Charlier G., Haesebrouck F. 1998; Typhlitis caused by intestinal Serpulina -like bacteria in domestic guinea pigs ( Cavia porcellus ). J Clin Microbiol 36:690–694
    [Google Scholar]
  40. Widjojoatmodjo M. N., Fluit A. C., Torensma R., Keller B. H., Verhoef J. 1991; Evaluation of the magnetic immuno PCR assay for rapid detection of Salmonella . Eur J Clin Microbiol Infect Dis 10:935–938 [CrossRef]
    [Google Scholar]
  41. Widjojoatmodjo M. N., Fluit A. C., Torensma R., Verdonk G. P., Verhoef J. 1992; The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples. J Clin Microbiol 30:3195–3199
    [Google Scholar]
  42. Windsor R. S., Simmons J. R. 1981; Investigations into the spread of swine dysentery in 25 herds in East Anglia and assessment of its economic significance in five herds. Vet Rec 109:482–484 [CrossRef]
    [Google Scholar]
  43. Wright D. J., Chapman P. A., Siddons C. A. 1994; Immunomagnetic separation as a sensitive method for isolating Escherichia coli O157 from food samples. Epidemiol Infect 113:31–39 [CrossRef]
    [Google Scholar]
  44. Zhang G., Weintraub A. 1998; Rapid and sensitivity assay for detection of enterotoxigenic Bacteroides fragilis . J Clin Microbiol 36:3545–3548
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.05500-0
Loading
/content/journal/jmm/10.1099/jmm.0.05500-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error