Journal Mobile Options
Table of Contents
Vol. 9, No. 1, 2005
Issue release date: 2005
J Mol Microbiol Biotechnol 2005;9:44–51

Isolation of Bacteriophages from Bartonella vinsonii subsp. berkhoffii and the Characterization of Pap31 Gene Sequences from Bacterial and Phage DNA

Maggi R.G. · Breitschwerdt E.B.
Vector-Borne Diseases Diagnostic Laboratory, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, N.C., USA

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Bacteriophages enhance bacterial survival, facilitate bacterial adaptation to new environmental conditions, assist in the adaptation to a new host species, and enhance bacterial evasion or inactivation of host defense mechanisms. We describe the detection and purification of a novel tailed bacteriophage from Bartonella vinsonii subsp. berkhoffii, which was previously described as a bacteriophage-negative species. We also compare B. vinsonii subsp. berkhoffi Pap31 bacteriophage gene sequences to B. henselae (Houston I), and B. quintana (Fuller) bacteriophage Pap31 sequences. Negative staining electron microscopy of log phase culturesof B. vinsonii subsp. berkhoffii identifiedbacteriophages, possessing a 50-nm icosahedric head diameter and a 60- to 80-nm contractile tail. Sequence analysis of the bacteriophage Pap31 gene from B. vinsonii subsp. berkhoffii showed three consensus sequences and a 12-bp insertion when compared with Pap31 gene sequences from B. henselae (Houston I) and B. quintana (Fuller) bacteriophages. Isolation of B. vinsonii subsp. berkhoffii bacteriophages containing a Pap31 gene suggests that this heme-binding protein gene might play an important role in bacterial virulence through the genetic exchange of DNA within this subspecies. Defining phage-associated genes may also contribute to the enhanced understanding of the evolutionary relationships among members of the genus Bartonella.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Adal KA, Cockerell CJ, Petri WA: Cat scratch disease, bacillary angiomatosis, and other infections due to Rochalimaea. N Engl J Med1994;330:1509–1515.
  2. Alsmark CA, Frank AC, Karleberg EO, Legault BA, Ardell DH, Canback B, Eriksson AS, Naslund AK, Handley SA, Huvet M, La Scola B, Holmberg M, Andersson SGE: The louse-borne human pathogen Bartonella quintana is a genomic derivative of the zoonotic agent Bartonella henselae. Proc Natl Acad Sci USA 2004;101:9716–9721.
  3. Anderson BE, Goldsmith C, Johnson A, Padmalayam I, Baumstark B: Bacteriophage-like particles of Rochalimaea henselae. Mol Microbiol 1994;13:67–73.
  4. Anderson BE, Neuman MA: Bartonella spp. as emerging human pathogens. Clin Microbiol Rev 1997a;10:203–219.
  5. Anderson BE, Scotchlas D, Jones D, Johnson A, Tzianabos T, Baumstark B: Analysis of a 36-kilodalton protein (PapA) associated with the bacteriophage particle of Bartonella henselae. DNA Cell Biol 1997b;16:1223–1229.
  6. Armengol CE, Hendley JO: Cat-scratch disease encephalopathy: a cause of status epilepticus in school-aged children. J Pediatr 1999;5:635–638.

    External Resources

  7. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K: Short Protocols in Molecular Biology, ed 3. New York, Wiley, 1997, pp 1512.
  8. Avidor B: Bartonella koehlera, a new human pathogen causing culture-negative endocarditis. 4th International Conference on Bartonella as Emerging Pathogens, Uppsala 2004.
  9. Banks DJ, Lei B, Musser JM: Prophage induction and expression of prophage-encoded virulence factors in group A Streptococcus serotype M3 strain MGAS315. Infect Immun 2003;71:7079–7086.
  10. Barbian KD, Minnick MF: A bacteriophage-like particle from Bartonella bacilliformis. Microbiology 2000;146:599–609.
  11. Barksdale L, Arden SB: Persisting bacteriophage infections, lysogeny, and phage conversions. Rev Microbiol 1974;28:265–299.
  12. Bergmans AMC, Schellekens JFP, Van Embden JDA, Schouls LM: Predominance of two Bartonella variants among cat scratch disease patients in The Netherlands. J Clin Microbiol 1996;34:254–260.
  13. Birge EA: Bacterial and Bacteriophage Genetics, ed 1. New York, Springer, 2000, pp 559.
  14. Birtles RJ, Canales J, Ventosilla P, Alvarez E, Guerra H, Llanos-Cuentas A, Raoult D, Doshi N, Harrison TG: Survey of Bartonella species infecting intradomicillary animals in the Huayllacallan Valley, Ancash, Peru, a region endemic for human bartonellosis. Am J Trop Med Hyg 1999;60:799–805.
  15. Birtles RJ, Fry NK, Ventosilla P, Caceres AG, Sanchez E, Vizcarra H, Raoult D: Identification of Bartonella bacilliformis genotypes and their relevance to epidemiological investigations of human bartonellosis. J Clin Microbiol 2002;40:3606–3612.
  16. Bishai WR, Murphy JR: Bacteriophage gene products that cause human disease; in Calendar R (ed): The Bacteriophages. New York, Plenum Press, 1988, pp 683–724.
  17. Bossi L, Figueroa-Bossi N: Inducible prophages contribute to Salmonella virulence in mice. Mol Microbiol 1999;33:167–176.
  18. Bossi L, Figueroa-Bossi N, Uzzau S, Maloriol D: Variable assortment of prophages provides a transferable repertoire of pathogenic determinants in Salmonella. Mol Microbiol 2001;39:260–271.
  19. Bowers TJ, Sweger D, Jue D, Anderson B: Isolation, sequencing and expression of the gene encoding a major protein from the bacteriophage-associated with Bartonella henselae. Gene 1998;206:49–52.
  20. Boyd EF, Davis BM, Hochhut B: Bacteriophage-bacteriophage interactions in the evolution of pathogenic bacteria. Trends Microbiol 2001;9:137–144.
  21. Boyd EF, Brussow H: Common themes among bacteriophage-encoded virulence factors and diversity among the bacteriophages involved. Trends Microbiol 2002;10:521–529.
  22. Breitschwerdt EB, Kordick DL, Malarkey DE: Endocarditis in a dog due to infection with a novel Bartonella subspecies. J Clin Microbiol 1995;33:154–160.
  23. Breitschwerdt EB, Atkins CE, Brown TT, Kordick DL, Snyder PS: Bartonellavinsonii subsp. berkhoffi and related members of the alpha subdivision of the Proteobacteria in dogs with cardiac arrhythmias, endocarditis, or myocarditis. J Clin Microbiol 1999;37:3618–3626.
  24. Breitschwerdt EB, Kordick DL: Bartonella infection in animals: carriership, reservoir potential, pathogenicity, and zoonotic potential for human infection. Clin Microbiol Rev 2000;13:428–438.
  25. Brouqui P, La Scola B, Roux V, Raoult D: Chronic Bartonella quintana bacteremia in homeless patients. N Engl J Med 1999;340:184–189.
  26. Carroll JA, Coleman SA, Smitherman LS, Mimmick M: Hemin-binding surface protein from Bartonella quintana. Infect Immun 2000;68:6750–6757.
  27. Chang CC, Chomel BB, Kasten RW, Heller R, Kocan KM, Ueno H, Yamamoto K, Bleich VC, Pierce BM, Gonzales BJ, Swift PK, Boyce WM, Jang SS, Boulouis HJ, Piémont Y: Bartonella spp. isolated from wild and domestic ruminants in North America. Emerg Infect Dis 2000;6:306–311.
  28. Cheetham BF, Katz ME: A role for bacteriophages in the evolution and transfer of bacterial virulence determinants. Mol Microbiol 1995;18:201–208.
  29. Chenoweth MR, Somerville GA, Krause DC, O’Reilly KL, Gherardini FC: Growth characteristics of Bartonella henselae in a novel liquid medium: primary isolation, growth-phase dependent phage induction, and metabolic studies. App Environ Microbiol 2004;70:656–663.
  30. Chomel BB, MacDonald KA, Kasten RW, Chang C, Wey AC, Foley JE, Thomas WP, Kittleson MD: Aortic valve endocarditis in a dog due to Bartonella clarridgeae. J Clin Microbiol 2001;39:3548–3554.
  31. Clarridge JE, Raich TJ, Pirwani D, Simon B, Tsai L, Rodriguez-Barradas MC, Regnery RL, Zollo A, Jones DC, Rambo C: Strategy to detect and identify Bartonella species in routine laboratory yields Bartonella henselae from human immunodeficiency virus-positive patient and unique Bartonella strain from his cat. J Clin Microbiol 1995;33:2107–2113.
  32. Daly JS, Worthington MG, Brenner DJ, Moss CW, Hollis DG, Weyant RS: Rochalimaea elizabethae sp. nov. isolated from a patient with endocarditis. J Clin Microbiol 1993;31:872–881.
  33. Dehio C: Bartonella interactions with endothelial cells and erythrocytes. Trends Microbiol 2001;9:279–285.
  34. Dehio C: Role of the type IV secretion system VirB/D4 in Bartonella pathogenesis. 4th International Conference on Bartonella as Emerging Pathogens, Uppsala 2004.
  35. DeLeo FR, Sturdevant DE, Braughtn KR, Kobayashi SD, Lei B, Virtaneva K, Dorward DW, Musser JM, Voyich JM: Genome-wide protective response used by group A Streptococcus to evade destruction by human polymorphonuclear leukocytes. Proc Natl Acad Sci USA2003;100:1996–2001.
  36. Drancourt M, Birtles R, Chaumentin G, Vandenesch F, Etienne J, Raoult D: New serotype of Bartonella henselae in endocarditis and cat-scratch disease. Lancet 1996;347:441–443.
  37. Ellis BA, Regnery RL, Beati L, Bacellar F, Rood M, Glass GG, Marston E, Ksiazek TG, Jones D, Childs JE: Rats of the genus Rattus are reservoir hosts for pathogenic Bartonella species: An old world origin for a new world disease? J Infect Dis 1999;180:220–224.
  38. Hadfield TL, Warren R, Kass M, Brun E, Levy C: Endocarditis caused by Rochalimaeahenselae. Hum Pathol 1993;24:1140–1141.
  39. Houpikian P, Raoult D: 16S/23S rRNA intergenic spacer regions for phylogenetic analysis, identification, and subtyping of Bartonella species. J Clin Microbiol 2001;39:2768–2778.
  40. Jacomo V, Kelly PJ, Raoult D: Natural history of Bartonella infections (an exception to Koch’s postulate). Clin Diagn Lab Immunol 2002;9:8–18.
  41. Jensen WA, Fall MZ, Rooney J, Kordick DL, Breitschwerdt EB: Rapid identification and differentiation of Bartonella species using a single-step PCR assay. J Clin Microbiol 2000;38:1717–1722.
  42. Joblet C, Roux V, Drancourt M, Gouvernet J, Raoult D: Identification of Bartonella(Rochalimaea) species among fastidious gram-negative bacteria on the basis of the partial sequence of the citrate-synthase gene. J Clin Microbiol 1995;33:1879–1883.
  43. Karem KL, Paddock CD, Regnery RL: Bartonella henselae, B. quintana, and B. bacilliformis: historical pathogens of emerging significance. Microbes Infect 2000;2:1193–1205.
  44. Kerkhoff FT, Bergmans AM, Van der Zee A, Rothova A: Demonstration of Bartonella grahamii DNA in ocular fluids of a patient with neuroretinitis. J Clin Microbiol 1999;37:4034–4038.
  45. Koehler JE, Quinn FD, Berger TG, LeBoit PE, Tappero JW: Isolation of Rochalimaea species from cutaneous and osseous lesions of bacillary angiomatosis. N Engl J Med 1992;327:1625–1631.
  46. Koehler JE: Clinical manifestation of Bartonella infections. 4th International Conference on Bartonella as Emerging Pathogens, Uppsala 2004.
  47. Kordick DL, Breitschwerdt EB: Intraerythrocytic presence of Bartonellahenselae. J Clin Microbiol 1995;33:1655–1656.
  48. Kordick DL, Swaminathan B, Greene CT, Wilson KH, O’Connor S, Hollis DG, Matar GM, Malcolm GB, Hayes PS, Hadfield TL, Breitschwerdt EB, Brenner DJ: Bartonellavinsonii subsp. berkhoffii subsp. nov., isolated from dogs, Bartonellavinsonii subsp. vinsonii, and emended description of Bartonellavinsonii. Int J Syst Bacteriol 1996;46:704–709.
  49. Kordick DL, Hilyard EJ, Hadfield TL, Wilson KH, Steigerwalt AG, Brenner DJ, Breitschwerdt EB: Bartonellaclarridgeiae, a newly recognized zoonotic pathogen causing inoculation papules, fever, and lymphadenopathy (cat scratch disease). J Clin Microbiol 1997;35:1813–1818.
  50. Maggi RG, Breitschwerdt EB: Bacteriophage in Bartonella vinsonii berkhoffii and characterization of its Pap31 Gene. 4th International Conference on Bartonella as Emerging Pathogens, Uppsala 2004.
  51. Maggi RG, Breitschwerdt EB: Potential limitations of the 16S-23S rRNA intergenic region for molecular detection of Bartonella species. J Clin Microbiol 2005;43:1171–1176.
  52. Maggi RG, Duncan AB, Breitschwerdt EB: Novel chemically defined liquid medium for the growth, as single or composed culture, and the primary isolation of Bartonella species from blood and body fluids. J Clin Microbiol 2005;43:2651–2655.
  53. Margileth AM, Hayden GF: Cat scratch disease – from feline affection to human infection. N Engl J Med 1993;329:53–54.
  54. Maurin M, Birtles R, Raoult D: Current knowledge of Bartonella species. Eur J Clin Microbiol Infect Dis 1997;16:487–506.
  55. Raoult D, Drancourt M, Carta A, Gastaut JA: Bartonella(Rochalimaea)quintana isolation in a patient with chronic adenopathy, lymphopenia, and a cat. Lancet 1994;343:977.
  56. Raoult D: Bartonella(Rochalimaea)quintana endocarditis in three homeless men. N Engl J Med 1995;332:419–423.
  57. Raoult D, Greub G, Fournier PE, Zeaiter Z: Diagnosis of Bartonella endocarditis by real-time nested PCR assay using serum. J Clin Microbiol 2003;41:919–925.
  58. Regnery RL, Anderson BE, Clarridge JE III, Rodriguez-Barradas M, Jones DC, Carr JH: Characterization of a novel Rochalimaea species, R. henselae sp. nov., isolated from blood of a febrile human immunodeficiency virus-positive patient. J Clin Microbiol 1992;30:265–274.
  59. Relman DA: Has trench fever returned? N Engl J Med1995;332:463–464.
  60. Schattner A, Zimhony O, Avidor B, Giladi M: Asking the right question. Lancet 2003;361:1786.
  61. Schmid MC, Schulein R, Dehio M, Denecker G, Carena I, Dehio C: The VirB type IV secretion system of Bartonella henselae mediates invasion, proinflammatory activation, and anti-apoptotic protection of endothelial cells. Mol Microbiol 2004;52:81–92.
  62. Schulein R, Dehio C: The VirB/VirD4 type IV secretion system of Bartonella is essential for establishing intraerythrocytic infection. Mol Microbiol 2002;46:1053–1067.
  63. Schuster SC, Lanz C, Raadatz G, Stockli M, Saenz H, Dehio C: Sequencing the Bartonella tribocorum genome. 4th International Conference on Bartonella as Emerging Pathogens, Uppsala 2004.
  64. Schwartzman W: Bartonella(Rochalimaea) infections: beyond cat scratch. Annu Rev Med 1996;47:355–364.
  65. Waldor MK: Bacteriophage biology and bacterial virulence. Trends Microbiol 1998;6:295–297.
  66. Welch DF, Pickett DA, Slater LN, Steigerwalt AG, Brenner DJ: Rochalimaea henselae sp. nov., a cause of septicemia, bacillary angiomatosis, and parenchymal bacillary peliosis. J Clin Microbiol 1992;30:275–280.
  67. Woestyn S, Olive N, Bigaignon G, Avesani V, Delmee M: Study of genotypes and virB4 secretion gene of Bartonella henselae strains from patients with clinically defined cat scratch disease. J Clin Microbiol 2004;42:1420–1427.
  68. Wong MT, Dolan MJ, Lattuada CP, Regenry RL, Garcia ML, Mokulis EC: Neuroretinitis, aseptic meningitis, and lymphadenitis associated with Bartonella henselae infection in immunocompetent patients and patients infected with human immunodeficiency virus type I. Clin Infect Dis 1995;21:352–360.
  69. Wright A: Mechanism of conversion of the Salmonella O antigen by bacteriophage epsilon 34. J Bacteriol 1971;105:927–936.
  70. Zimmermann R, Kempf VAJ, Schiltz E, Oberle K, Sander A: Hemin binding, functional expression, and complementation analysis of Pap31 from Bartonella henselae. J Bacteriol 2003;185:1739–1744.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50