Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Mar;34(3):628–633. doi: 10.1128/jcm.34.3.628-633.1996

Determination of stability of Brucella abortus RB51 by use of genomic fingerprint, oxidative metabolism, and colonial morphology and differentiation of strain RB51 from B. abortus isolates from bison and elk.

A E Jensen 1, D R Ewalt 1, N F Cheville 1, C O Thoen 1, J B Payeur 1
PMCID: PMC228859  PMID: 8904427

Abstract

Brucella abortus RB51 and isolates from cattle, bison, and elk were characterized by pulsed-field gel electrophoresis and standard techniques for biotyping Brucella species, which included biochemical, morphological, and antigenic techniques, phage susceptibility, and antibiotic resistance. The objectives were to ascertain the stability of RB51 and to differentiate RB51 from other brucellae. Genomic restriction endonuclease patterns produced by pulsed-field gel electrophoresis demonstrated a unique fingerprint for RB51 relative to other brucellae. Comparisons of the oxidative metabolic profiles of RB51 after time in vivo (14 weeks) and in vitro (75 passages) showed no change in characteristic patterns of oxygen uptake on selected amino acid and carbohydrate substrates. Strain RB51 was biotyped as a typical rough B. abortus biovar 1 (not strain 19) after animal passage or a high number of passages in vitro and remained resistant to rifampin or penicillin and susceptible to tetracycline. No reactions with A or M antiserum or with a monoclonal antibody to the O antigen of Brucella lipopolysaccharides were detected; however, RB51 agglutinated with R antiserum. The results indicate that the genomic fingerprint and rough colonial morphology of RB51 are stable characteristics and can be used to differentiate this vaccine strain from Brucella isolates from cattle, bison, and elk.

Full Text

The Full Text of this article is available as a PDF (329.6 KB).

Selected References

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

  1. Allardet-Servent A., Bourg G., Ramuz M., Pages M., Bellis M., Roizes G. DNA polymorphism in strains of the genus Brucella. J Bacteriol. 1988 Oct;170(10):4603–4607. doi: 10.1128/jb.170.10.4603-4607.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bauer A. W., Kirby W. M., Sherris J. C., Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966 Apr;45(4):493–496. [PubMed] [Google Scholar]
  3. Cheville N. F., Jensen A. E., Halling S. M., Tatum F. M., Morfitt D. C., Hennager S. G., Frerichs W. M., Schurig G. Bacterial survival, lymph node changes, and immunologic responses of cattle vaccinated with standard and mutant strains of Brucella abortus. Am J Vet Res. 1992 Oct;53(10):1881–1888. [PubMed] [Google Scholar]
  4. Cheville N. F., Jensen A. E., Morfitt D. C., Stabel T. J. Cutaneous delayed hypersensitivity reactions of cattle vaccinated with mutant strains of Brucella abortus, using brucellins prepared from various brucellar strains. Am J Vet Res. 1994 Sep;55(9):1261–1266. [PubMed] [Google Scholar]
  5. Cheville N. F., Stevens M. G., Jensen A. E., Tatum F. M., Halling S. M. Immune responses and protection against infection and abortion in cattle experimentally vaccinated with mutant strains of Brucella abortus. Am J Vet Res. 1993 Oct;54(10):1591–1597. [PubMed] [Google Scholar]
  6. Corbel M. J. Determination of the in vitro sensitivity of Brucella strains to rifampicin. Br Vet J. 1976 May-Jun;132(3):266–275. doi: 10.1016/s0007-1935(17)34686-9. [DOI] [PubMed] [Google Scholar]
  7. Farrell I. D. The development of a new selective medium for the isolation of Brucella abortus from contaminated sources. Res Vet Sci. 1974 May;16(3):280–286. [PubMed] [Google Scholar]
  8. Feingold D. S., HsuChen C. C., Sud I. J. Basis for the selectivity of action of the polymyxin antibiotics on cell membranes. Ann N Y Acad Sci. 1974 May 10;235(0):480–492. doi: 10.1111/j.1749-6632.1974.tb43285.x. [DOI] [PubMed] [Google Scholar]
  9. Gamazo C., Moriyón I. Release of outer membrane fragments by exponentially growing Brucella melitensis cells. Infect Immun. 1987 Mar;55(3):609–615. doi: 10.1128/iai.55.3.609-615.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jensen A. E., Cheville N. F., Ewalt D. R., Payeur J. B., Thoen C. O. Application of pulsed-field gel electrophoresis for differentiation of vaccine strain RB51 from field isolates of Brucella abortus from cattle, bison, and elk. Am J Vet Res. 1995 Mar;56(3):308–312. [PubMed] [Google Scholar]
  11. MEYER M. E., CAMERON H. S. Metabolic characterization of the genus Brucella. I. Statistical evaluation of the oxidative rates by which type I of each species can be identified. J Bacteriol. 1961 Sep;82:387–395. doi: 10.1128/jb.82.3.387-395.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meyer M. E. Characterization of Brucella abortus strain 19 isolated from human and bovine tissues and fluids. Am J Vet Res. 1985 Apr;46(4):902–904. [PubMed] [Google Scholar]
  13. Moore C. G., Schnurrenberger P. R. A review of naturally occurring Brucella abortus infections in wild mammals. J Am Vet Med Assoc. 1981 Dec 1;179(11):1105–1112. [PubMed] [Google Scholar]
  14. Moreno E., Berman D. T., Boettcher L. A. Biological activities of Brucella abortus lipopolysaccharides. Infect Immun. 1981 Jan;31(1):362–370. doi: 10.1128/iai.31.1.362-370.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Moriyon I., Berman D. T. Effects of nonionic, ionic, and dipolar ionic detergents and EDTA on the Brucella cell envelope. J Bacteriol. 1982 Nov;152(2):822–828. doi: 10.1128/jb.152.2.822-828.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Phillips M., Deyoe B. L., Canning P. C. Protection of mice against Brucella abortus infection by inoculation with monoclonal antibodies recognizing Brucella O-antigen. Am J Vet Res. 1989 Dec;50(12):2158–2161. [PubMed] [Google Scholar]
  17. Rhyan J. C., Quinn W. J., Stackhouse L. S., Henderson J. J., Ewalt D. R., Payeur J. B., Johnson M., Meagher M. Abortion caused by Brucella abortus biovar 1 in a free-ranging bison (Bison bison) from Yellowstone National Park. J Wildl Dis. 1994 Jul;30(3):445–446. doi: 10.7589/0090-3558-30.3.445. [DOI] [PubMed] [Google Scholar]
  18. Rifkind D. Studies on the interaction between endotoxin and polymyxin B. J Infect Dis. 1967 Dec;117(5):433–438. doi: 10.1093/infdis/117.5.433. [DOI] [PubMed] [Google Scholar]
  19. Roop R. M., 2nd, Jeffers G., Bagchi T., Walker J., Enright F. M., Schurig G. G. Experimental infection of goat fetuses in utero with a stable, rough mutant of Brucella abortus. Res Vet Sci. 1991 Sep;51(2):123–127. doi: 10.1016/0034-5288(91)90001-5. [DOI] [PubMed] [Google Scholar]
  20. Schurig G. G., Roop R. M., 2nd, Bagchi T., Boyle S., Buhrman D., Sriranganathan N. Biological properties of RB51; a stable rough strain of Brucella abortus. Vet Microbiol. 1991 Jul;28(2):171–188. doi: 10.1016/0378-1135(91)90091-s. [DOI] [PubMed] [Google Scholar]
  21. Schwartz D. C., Cantor C. R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. doi: 10.1016/0092-8674(84)90301-5. [DOI] [PubMed] [Google Scholar]
  22. Stevens M. G., Hennager S. G., Olsen S. C., Cheville N. F. Serologic responses in diagnostic tests for brucellosis in cattle vaccinated with Brucella abortus 19 or RB51. J Clin Microbiol. 1994 Apr;32(4):1065–1066. doi: 10.1128/jcm.32.4.1065-1066.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stevens M. G., Olsen S. C., Cheville N. F. Comparative analysis of immune responses in cattle vaccinated with Brucella abortus strain 19 or strain RB51. Vet Immunol Immunopathol. 1995 Feb;44(3-4):223–235. doi: 10.1016/0165-2427(94)05311-f. [DOI] [PubMed] [Google Scholar]
  24. Terakado N., Ueda H., Sugawara H., Isayama Y., Koyama N. Drug susceptibility of Brucella canis isolated from dogs. Nihon Juigaku Zasshi. 1978 Jun;40(3):291–295. doi: 10.1292/jvms1939.40.291. [DOI] [PubMed] [Google Scholar]
  25. Tessaro S. V. The existing and potential importance of brucellosis and tuberculosis in canadian wildlife: a review. Can Vet J. 1986 Mar;27(3):119–124. [PMC free article] [PubMed] [Google Scholar]
  26. Verger J. M., Grayon M. Oxidative metabolic profiles of "Brucella" species. Ann Sclavo. 1977 Jan-Feb;19(1):46–60. [PubMed] [Google Scholar]
  27. WHITE P. G., WILSON J. B. Differentiation of smooth and nonsmooth colonies of Brucellae. J Bacteriol. 1951 Feb;61(2):239–240. doi: 10.1128/jb.61.2.239-240.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Williams E. S., Thorne E. T., Anderson S. L., Herriges J. D., Jr Brucellosis in free-ranging bison (Bison bison) from Teton County, Wyoming. J Wildl Dis. 1993 Jan;29(1):118–122. doi: 10.7589/0090-3558-29.1.118. [DOI] [PubMed] [Google Scholar]

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

RESOURCES