Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Oct;34(10):2460–2463. doi: 10.1128/jcm.34.10.2460-2463.1996

Molecular characterization of Salmonella enterica subsp. enterica serovar choleraesuis field isolates and differentiation from homologous live vaccine strains suisaloral and SC-54.

M Weide-Botjes 1, B Liebisch 1, S Schwarz 1, J L Watts 1
PMCID: PMC229294  PMID: 8880500

Abstract

Four independent molecular methods were used to characterize the Salmonella enterica subsp. enterica serovar choleraesuis live vaccine strains SC-54 and Suisaloral and to differentiate them from S. choleraesuis field isolates. Plasmid analysis revealed the presence of seven plasmid profiles. A virulence plasmid of 52-kbp was identified by hybridization with an spvB-spvC gene probe in each of the S. choleraesuis field isolates and in the Suisaloral vaccine strain, but not in the SC-54 vaccine strain. Ribotyping, performed with a gene probe that recognized 23S, 16S, and 5S rRNA genes, resulted in three closely related hybridization patterns. IS200 elements were not detected in the field isolates or in the two S. choleraesuis live vaccine strains. Macrorestriction analysis with the enzymes XbaI, SpeI, NotI, and SfiI differentiated the 29 S. choleraesuis strains included in this study into 10, 13, 8, and 13 different fragment patterns, respectively. While the Suisaloral vaccine strain showed a unique XbaI macrorestriction pattern, the fragment patterns of the SC-54 strain obtained with the different enzymes were shared by 2 to 18 S. choleraesuis field strains. A combination of plasmid analysis and macrorestriction analysis proved to be most suitable for the molecular typing of S. choleraesuis and the differentiation of both live vaccine strains from field isolates of this serovar.

Full Text

The Full Text of this article is available as a PDF (255.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beyer W., Geue L. Characterization of the virulence regions in the plasmids of three live Salmonella vaccines. Zentralbl Bakteriol. 1992 Jun;277(1):10–21. doi: 10.1016/s0934-8840(11)80865-0. [DOI] [PubMed] [Google Scholar]
  2. Brosius J., Ullrich A., Raker M. A., Gray A., Dull T. J., Gutell R. R., Noller H. F. Construction and fine mapping of recombinant plasmids containing the rrnB ribosomal RNA operon of E. coli. Plasmid. 1981 Jul;6(1):112–118. doi: 10.1016/0147-619x(81)90058-5. [DOI] [PubMed] [Google Scholar]
  3. Chowdry N., Threlfall E. J., Rowe B., Stanley J. Genotype analysis of faecal and blood isolates of Salmonella dublin from humans in England and Wales. Epidemiol Infect. 1993 Apr;110(2):217–225. doi: 10.1017/s0950268800068138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Esteban E., Snipes K., Hird D., Kasten R., Kinde H. Use of ribotyping for characterization of Salmonella serotypes. J Clin Microbiol. 1993 Feb;31(2):233–237. doi: 10.1128/jcm.31.2.233-237.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gibert I., Barbé J., Casadesús J. Distribution of insertion sequence IS200 in Salmonella and Shigella. J Gen Microbiol. 1990 Dec;136(12):2555–2560. doi: 10.1099/00221287-136-12-2555. [DOI] [PubMed] [Google Scholar]
  6. Gulig P. A. Virulence plasmids of Salmonella typhimurium and other salmonellae. Microb Pathog. 1990 Jan;8(1):3–11. doi: 10.1016/0882-4010(90)90003-9. [DOI] [PubMed] [Google Scholar]
  7. Hunter P. R., Gaston M. A. Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J Clin Microbiol. 1988 Nov;26(11):2465–2466. doi: 10.1128/jcm.26.11.2465-2466.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kado C. I., Liu S. T. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. doi: 10.1128/jb.145.3.1365-1373.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kramer T. T., Roof M. B., Matheson R. R. Safety and efficacy of an attenuated strain of Salmonella choleraesuis for vaccination of swine. Am J Vet Res. 1992 Apr;53(4):444–448. [PubMed] [Google Scholar]
  10. Liebisch B., Schwarz S. Evaluation and comparison of molecular techniques for epidemiological typing of Salmonella enterica subsp. enterica serovar dublin. J Clin Microbiol. 1996 Mar;34(3):641–646. doi: 10.1128/jcm.34.3.641-646.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Liebisch B., Schwarz S. Molecular typing of Salmonella enterica subsp. enterica serovar Enteritidis isolates. J Med Microbiol. 1996 Jan;44(1):52–59. doi: 10.1099/00222615-44-1-52. [DOI] [PubMed] [Google Scholar]
  12. Macrina F. L., Kopecko D. J., Jones K. R., Ayers D. J., McCowen S. M. A multiple plasmid-containing Escherichia coli strain: convenient source of size reference plasmid molecules. Plasmid. 1978 Jun;1(3):417–420. doi: 10.1016/0147-619x(78)90056-2. [DOI] [PubMed] [Google Scholar]
  13. Millemann Y., Lesage M. C., Chaslus-Dancla E., Lafont J. P. Value of plasmid profiling, ribotyping, and detection of IS200 for tracing avian isolates of Salmonella typhimurium and S. enteritidis. J Clin Microbiol. 1995 Jan;33(1):173–179. doi: 10.1128/jcm.33.1.173-179.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Montenegro M. A., Morelli G., Helmuth R. Heteroduplex analysis of Salmonella virulence plasmids and their prevalence in isolates of defined sources. Microb Pathog. 1991 Dec;11(6):391–397. doi: 10.1016/0882-4010(91)90035-9. [DOI] [PubMed] [Google Scholar]
  15. Olsen J. E., Brown D. J., Skov M. N., Christensen J. P. Bacterial typing methods suitable for epidemiological analysis. Applications in investigations of salmonellosis among livestock. Vet Q. 1993 Dec;15(4):125–135. doi: 10.1080/01652176.1993.9694390. [DOI] [PubMed] [Google Scholar]
  16. Olsen J. E., Skov M. N., Threlfall E. J., Brown D. J. Clonal lines of Salmonella enterica serotype Enteritidis documented by IS200-, ribo-, pulsed-field gel electrophoresis and RFLP typing. J Med Microbiol. 1994 Jan;40(1):15–22. doi: 10.1099/00222615-40-1-15. [DOI] [PubMed] [Google Scholar]
  17. Olsen J. E., Skov M. Genomic lineage of Salmonella enterica serovar Dublin. Vet Microbiol. 1994 Jun;40(3-4):271–282. doi: 10.1016/0378-1135(94)90116-3. [DOI] [PubMed] [Google Scholar]
  18. Roof M. B., Doitchinoff D. D. Safety, efficacy, and duration of immunity induced in swine by use of an avirulent live Salmonella choleraesuis-containing vaccine. Am J Vet Res. 1995 Jan;56(1):39–44. [PubMed] [Google Scholar]
  19. Schwarz S., Liebisch B. Pulsed-field gel electrophoretic identification of Salmonella enterica serovar Typhimurium live vaccine strain Zoosaloral H. Lett Appl Microbiol. 1994 Dec;19(6):469–472. doi: 10.1111/j.1472-765x.1994.tb00984.x. [DOI] [PubMed] [Google Scholar]
  20. Schwarz S., Liebisch B. Use of ribotyping, IS200 typing and plasmid analysis for the identification of Salmonella enterica subsp. enterica serovar Typhimurium vaccine strain Zoosaloral H and its differentiation from wild type strains of the same serovar. Zentralbl Bakteriol. 1994 Nov;281(4):442–450. doi: 10.1016/s0934-8840(11)80330-0. [DOI] [PubMed] [Google Scholar]
  21. Schöll W., Grünert G. Suissaloral "Dessau"--ein Salmonella-cholerae-suis-Lebendimpfstoff zur oralen, parenteralen und kombinierten Anwendung. Arch Exp Veterinarmed. 1980;34(1):91–97. [PubMed] [Google Scholar]
  22. Stanley J., Baquar N., Threlfall E. J. Genotypes and phylogenetic relationships of Salmonella typhimurium are defined by molecular fingerprinting of IS200 and 16S rrn loci. J Gen Microbiol. 1993 Jun;139(Pt 6):1133–1140. doi: 10.1099/00221287-139-6-1133. [DOI] [PubMed] [Google Scholar]
  23. Stanley J., Burnens A. P., Threlfall E. J., Chowdry N., Goldsworthy M. Genetic relationships among strains of Salmonella enteritidis in a national epidemic in Switzerland. Epidemiol Infect. 1992 Apr;108(2):213–220. doi: 10.1017/s0950268800049694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tenover F. C., Arbeit R. D., Goering R. V., Mickelsen P. A., Murray B. E., Persing D. H., Swaminathan B. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995 Sep;33(9):2233–2239. doi: 10.1128/jcm.33.9.2233-2239.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Thong K. L., Ngeow Y. F., Altwegg M., Navaratnam P., Pang T. Molecular analysis of Salmonella enteritidis by pulsed-field gel electrophoresis and ribotyping. J Clin Microbiol. 1995 May;33(5):1070–1074. doi: 10.1128/jcm.33.5.1070-1074.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Threlfall E. J., Frost J. A. The identification, typing and fingerprinting of Salmonella: laboratory aspects and epidemiological applications. J Appl Bacteriol. 1990 Jan;68(1):5–16. doi: 10.1111/j.1365-2672.1990.tb02542.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES