Europe PMC

This website requires cookies, and the limited processing of your personal data in order to function. By using the site you are agreeing to this as outlined in our privacy notice and cookie policy.

Isolation, cultivation, and partial characterization of the ELB agent associated with cat fleas.

Author information

Affiliations

  • 1. Department of Microbiology and Immunology, School of Medicine, University of Baltimore, Maryland 21201-1559, USA.
      Authors
    • Radulovic S 1
    (1 author)

Infection and Immunity, 01 Dec 1995, 63(12):4826-4829
https://doi.org/10.1128/iai.63.12.4826-4829.1995 PMID: 7591142 PMCID: PMC173691

Free full text in Europe PMC

Abstract 

ELB rickettsiae from cat flea homogenates were recovered in tissue culture cells following sequential passage through laboratory rats and the yolk sacs of embryonated chicken eggs. Seven days after inoculation of ELB from the infected yolk sacs, Vero cells and L929 cells were observed to contain intracellular bacteria as demonstrated by Diff Quik and indirect immunofluorescence assay staining. The rickettsial and ELB identity of the cultured agent was confirmed by PCR detection of the 16S rRNA and citrate synthase genes and PCR-restriction fragment length polymorphism analysis of the 17-kDa conserved rickettsial antigen gene. The ELB rickettsiae induced plaques in Vero cells on day 11 postinfection. Rat anti-ELB serum reacted at 1:4,096 to cultured ELB and had lower reactivity to Rickettsia typhi Wilmington (1:1,024), Rickettsia akari Kaplan (1:512), and Rickettsia australis JC (1:64). Spotted fever group polyclonal sera also exhibited lower reactivity to ELB than to the homologous antigen. Coomassie blue-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of the ELB isolate and two R. typhi strains were identical.

Free full text 

Logo of iaiLink to Publisher's site
Infect Immun. 1995 Dec; 63(12): 4826–4829.
PMCID: PMC173691
PMID: 7591142

Isolation, cultivation, and partial characterization of the ELB agent associated with cat fleas.

S Radulovic, J A Higgins, D C Jaworski, G A Dasch, and A F Azad
Author information Copyright and License information Disclaimer

Abstract

ELB rickettsiae from cat flea homogenates were recovered in tissue culture cells following sequential passage through laboratory rats and the yolk sacs of embryonated chicken eggs. Seven days after inoculation of ELB from the infected yolk sacs, Vero cells and L929 cells were observed to contain intracellular bacteria as demonstrated by Diff Quik and indirect immunofluorescence assay staining. The rickettsial and ELB identity of the cultured agent was confirmed by PCR detection of the 16S rRNA and citrate synthase genes and PCR-restriction fragment length polymorphism analysis of the 17-kDa conserved rickettsial antigen gene. The ELB rickettsiae induced plaques in Vero cells on day 11 postinfection. Rat anti-ELB serum reacted at 1:4,096 to cultured ELB and had lower reactivity to Rickettsia typhi Wilmington (1:1,024), Rickettsia akari Kaplan (1:512), and Rickettsia australis JC (1:64). Spotted fever group polyclonal sera also exhibited lower reactivity to ELB than to the homologous antigen. Coomassie blue-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of the ELB isolate and two R. typhi strains were identical.

Full Text

The Full Text of this article is available as a PDF (288K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adams JR, Schmidtmann ET, Azad AF. Infection of colonized cat fleas, Ctenocephalides felis (Bouché), with a rickettsia-like microorganism. Am J Trop Med Hyg. 1990 Oct;43(4):400–409. [Abstract] [Google Scholar]
  • Adams WH, Emmons RW, Brooks JE. The changing ecology of murine (endemic) typhus in Southern California. Am J Trop Med Hyg. 1970 Mar;19(2):311–318. [Abstract] [Google Scholar]
  • Azad AF, Sacci JB, Jr, Nelson WM, Dasch GA, Schmidtmann ET, Carl M. Genetic characterization and transovarial transmission of a typhus-like rickettsia found in cat fleas. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):43–46. [Europe PMC free article] [Abstract] [Google Scholar]
  • Beati L, Finidori JP, Gilot B, Raoult D. Comparison of serologic typing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein analysis, and genetic restriction fragment length polymorphism analysis for identification of rickettsiae: characterization of two new rickettsial strains. J Clin Microbiol. 1992 Aug;30(8):1922–1930. [Europe PMC free article] [Abstract] [Google Scholar]
  • Boman HG, Faye I, Gudmundsson GH, Lee JY, Lidholm DA. Cell-free immunity in Cecropia. A model system for antibacterial proteins. Eur J Biochem. 1991 Oct 1;201(1):23–31. [Abstract] [Google Scholar]
  • Cociancich S, Bulet P, Hetru C, Hoffmann JA. The inducible antibacterial peptides of insects. Parasitol Today. 1994 Apr;10(4):132–139. [Abstract] [Google Scholar]
  • Daffre S, Kylsten P, Samakovlis C, Hultmark D. The lysozyme locus in Drosophila melanogaster: an expanded gene family adapted for expression in the digestive tract. Mol Gen Genet. 1994 Jan;242(2):152–162. [Abstract] [Google Scholar]
  • Gilmore RD, Jr, Cieplak W, Jr, Policastro PF, Hackstadt T. The 120 kilodalton outer membrane protein (rOmp B) of Rickettsia rickettsii is encoded by an unusually long open reading frame: evidence for protein processing from a large precursor. Mol Microbiol. 1991 Oct;5(10):2361–2370. [Abstract] [Google Scholar]
  • Gilmore RD, Jr, Hackstadt T. DNA polymorphism in the conserved 190 kDa antigen gene repeat region among spotted fever group Rickettsiae. Biochim Biophys Acta. 1991 Jul 26;1097(1):77–80. [Abstract] [Google Scholar]
  • Hoffmann JA, Hetru C. Insect defensins: inducible antibacterial peptides. Immunol Today. 1992 Oct;13(10):411–415. [Abstract] [Google Scholar]
  • Hultmark D. Immune reactions in Drosophila and other insects: a model for innate immunity. Trends Genet. 1993 May;9(5):178–183. [Abstract] [Google Scholar]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [Abstract] [Google Scholar]
  • Philip RN, Casper EA, Burgdorfer W, Gerloff RK, Hughes LE, Bell EJ. Serologic typing of rickettsiae of the spotted fever group by microimmunofluorescence. J Immunol. 1978 Nov;121(5):1961–1968. [Abstract] [Google Scholar]
  • Raoult D, Dasch GA. The line blot: an immunoassay for monoclonal and other antibodies. Its application to the serotyping of gram-negative bacteria. J Immunol Methods. 1989 Dec 20;125(1-2):57–65. [Abstract] [Google Scholar]
  • Regnery RL, Spruill CL, Plikaytis BD. Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes. J Bacteriol. 1991 Mar;173(5):1576–1589. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schriefer ME, Sacci JB, Jr, Dumler JS, Bullen MG, Azad AF. Identification of a novel rickettsial infection in a patient diagnosed with murine typhus. J Clin Microbiol. 1994 Apr;32(4):949–954. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schriefer ME, Sacci JB, Jr, Taylor JP, Higgins JA, Azad AF. Murine typhus: updated roles of multiple urban components and a second typhuslike rickettsia. J Med Entomol. 1994 Sep;31(5):681–685. [Abstract] [Google Scholar]
  • Sorvillo FJ, Gondo B, Emmons R, Ryan P, Waterman SH, Tilzer A, Andersen EM, Murray RA, Barr R. A suburban focus of endemic typhus in Los Angeles County: association with seropositive domestic cats and opossums. Am J Trop Med Hyg. 1993 Feb;48(2):269–273. [Abstract] [Google Scholar]
  • Weiss E, Coolbaugh JC, Williams JC. Separation of viable Rickettsia typhi from yolk sac and L cell host components by renografin density gradient centrifugation. Appl Microbiol. 1975 Sep;30(3):456–463. [Europe PMC free article] [Abstract] [Google Scholar]
  • Werren JH, Hurst GD, Zhang W, Breeuwer JA, Stouthamer R, Majerus ME. Rickettsial relative associated with male killing in the ladybird beetle (Adalia bipunctata). J Bacteriol. 1994 Jan;176(2):388–394. [Europe PMC free article] [Abstract] [Google Scholar]
  • Williams SG, Sacci JB, Jr, Schriefer ME, Andersen EM, Fujioka KK, Sorvillo FJ, Barr AR, Azad AF. Typhus and typhuslike rickettsiae associated with opossums and their fleas in Los Angeles County, California. J Clin Microbiol. 1992 Jul;30(7):1758–1762. [Europe PMC free article] [Abstract] [Google Scholar]
Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

Full text links 

Read article at publisher's site: https://doi.org/10.1128/iai.63.12.4826-4829.1995

Read article for free, from open access legal sources, via Unpaywall: https://iai.asm.org/content/iai/63/12/4826.full.pdfUnpaywall

Free after 4 months at iai.asm.org
http://iai.asm.org/cgi/reprint/63/12/4826

Free to read at iai.asm.org
http://iai.asm.org/cgi/content/abstract/63/12/4826

Citations & impact 

Impact metrics

26
Citations
Jump to Citations

Citations of article over time

Article citations

Go to all (26) article citations

Similar Articles 

To arrive at the top five similar articles we use a word-weighted algorithm to compare words from the Title and Abstract of each citation.

Funding 

Funders who supported this work.

NIAID NIH HHS (1)